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Biomedical subjects

D Levin

Publications and source records attributed to D Levin.

At least 37 records · Page 2Linked to original sources
PubMed1996

Modified thioglycolate medium: a simple and reliable means for detection of Trichomonas vaginalis.

Despite the declining rate of sexually transmitted diseases in developed countries, trichomoniasis is still one of the most common venereal infections. While diagnosis of this condition is commonly based on the microscopic wet-mount method, culture remains the most accurate single procedure for detecting the presence of Trichomonas vaginalis in clinical samples. In the present study, the efficacy of a modified formula of the commonly available thioglycolate medium was compared with that of the standard Diamond's medium for detection of T. vaginalis in samples from 176 women with vaginal symptoms. Thioglycolate medium supplemented with yeast extract, horse serum, and antimicrobial agents was as reliable as Diamond's medium for detection of T. vaginalis in vaginal fluid samples. Modified thioglycolate medium may be used as a readily available, low-cost substitute for the standard medium for culturing T. vaginalis.

Animals↗
PubMed1996

Expression of the inducible nitric oxide synthase gene in diaphragm and skeletal muscle.

Nitric oxide (NO) is a pluripotent molecule that can be secreted by skeletal muscle through the activity of the neuronal constitutive isoform of NO synthase. To determine whether skeletal muscle and diaphragm might also express the macrophage-inducible form of NO synthase (iNOS) during provocative states, we examined tissue from mice at serial times after intravenous administration of Escherichia coli endotoxin. In these studies, iNOS mRNA was strongly expressed in the diaphragm and skeletal muscle of mice 4 h after intravenous endotoxin and was significantly diminished by 8 h after challenge. Induction of iNOS mRNA was followed by expression of iNOS immunoreactive protein on Western immunoblots. Increased iNOS activity was demonstrated by conversion of arginine to citrulline. Immunochemical analysis of diaphragmatic explants exposed to endotoxin in vitro revealed specific iNOS staining in myocytes, in addition to macrophages and endothelium. These results may be important in understanding the pathogenesis of respiratory pump failure during septic shock, as well as skeletal muscle injury during inflammation or metabolic stress.

Animals↗
PubMed1995

Peptide and protein antigens require distinct antigen-presenting cell subsets for the priming of CD4+ T cells.

Priming of naive CD4+ T cells to Ag requires an antigen-presenting cell (APC) that can take up the Ag and present peptide bound to MHC class II molecules. We have used both in vivo and in vitro approaches to demonstrate that the APC used to prime naive CD4+ T cells depends on the initial form in which an Ag is administered. Although Ag delivered as a peptide was presented most efficiently to CD4+ T cells by DC, these APC were poor at priming to a protein form of the same Ag. In contrast, the presence of B cells was a requisite for priming to protein Ag.

Animals↗
PubMed1995

Regulation of transgenic class II major histocompatibility genes in murine Langerhans cells.

I-E is a class II major histocompatibility complex molecule normally expressed by Langerhans cells. A series of transgenic mice were developed previously that carry E alpha d gene constructs with promoter-region deletions that cause expression of I-E by different cell types when maintained on a B6 (I-E[-]) genetic background. To study cis-acting gene sequences that regulate expression of class II proteins by Langerhans cells, we identified transgenic I-E expression by tissue immunoperoxidase staining and by epidermal cell suspension immunofluorescence cytometry. Mice with a transgene containing 1.4 kilobase pairs (kb) of flanking sequence 5' to the E alpha initiation site expressed barely detectable levels of I-E on a tiny percentage of Langerhans cells, indicating that sequences promoting Langerhans cell expression of E alpha exist between 2.0 and 1.4 kb 5' of the E alpha initiation site. Removal of an additional 170 bp of 5' flanking sequence caused near-normal levels of expression by approximately one third of epidermal Langerhans cells, which contrasts with studies that showed minimal transgene expression by splenic dendritic cells in these animals. Thus, sequences between 1.4 and 1.23 kb 5' of the E alpha initiation site decrease expression of I-E by epidermal Langerhans cells, but enable I-E expression by splenic dendritic cells. These studies identify Langerhans cell-specific regulatory sequences and genetic regions controlling major histocompatibility complex class II gene expression in Langerhans cells and splenic dendritic cells. The genetic regions identified may be particularly important because differential regulation of class II major histocompatibility complex protein synthesis by Langerhans cells and dendritic cells may be crucial to immune functions of intact animals.

Animals↗
PubMed1994

Brain death in pediatric intensive care unit patients: incidence, primary diagnosis, and the clinical occurrence of Turner's triad.

OBJECTIVES: To determine the incidence and characteristics of children with brain death in the pediatric intensive care unit (ICU), and to assess the incidence of the clinical triad (Turner's triad) of central diabetes insipidus, low glucose demand, and low CO2 production. DESIGN: Retrospective review of medical records. SETTING: Two multidisciplinary pediatric ICUs. PATIENTS: Medical records of pediatric patients declared brain dead and admitted to two multidisciplinary pediatric ICUs. MEASUREMENTS AND MAIN RESULTS: Patient records were reviewed for demographic data, primary diagnosis, severity of illness (Pediatric Risk of Mortality score and calculated risk of mortality), methods by which brain-death diagnosis was determined, presence of central diabetes insipidus, low glucose demand, low CO2 production in the final 24 hrs before the diagnosis was made, and whether organ donation was accomplished. The incidence of brain death among all patients admitted to the pediatric ICUs was 0.9%, accounting for 11% of patients who died during the same period. The most common presentation leading to brain-death diagnoses was trauma, followed by drowning/near drowning, and meningitis. The majority of brain-death diagnoses were made using both clinical criteria and confirmatory tests (66%). The incidence of clinical signs of Turner's triad was 41% for central diabetes insipidus, 49% for low glucose demand, and 53% for low CO2 production. Two of the three features were present in 38% of patients, and 12% of the patients had all three features. CONCLUSIONS: Our series of brain-dead patients in the pediatric ICU showed a 0.9% incidence of brain death. The most common primary diagnosis was trauma, a finding that is similar to other series. We also demonstrated that the clinical triad (Turner's triad) is present in this patient population, although only 12% of study patients demonstrated all three features.

Adolescent↗
PubMed1994

Carcinoma of the prostate gland: MR imaging with pelvic phased-array coils versus integrated endorectal--pelvic phased-array coils.

PURPOSE: To compare the performance of pelvic phased-array (PPA) coils and integrated endorectal PPA coils in evaluation of local extent of prostate cancer. MATERIALS AND METHODS: Seventy-one men underwent magnetic resonance (MR) imaging within 3 weeks of radical prostatectomy. MR findings of tumor, extracapsular extension, seminal vesicle invasion, nodal involvement, and stage were compared with step-sectioned (3-4-mm intervals) whole-mounted pathologic specimens. Prospective consensus versus single-reader interpretation (kappa statistics) and PPA versus integrated endorectal PPA coils (McNemar test) were compared, and a rating scale of 1-6 was developed for analysis of receiver operating characteristics. Statistical significance was calculated at delta = .05. RESULTS: Comparison between consensus and single-reader image interpretation showed positive but poor agreement (kappa = .38) and no statistical significance. Staging accuracy was better (difference approached significance) for integrated endorectal PPA coils (77%) than for PPA coils (68%). CONCLUSION: The integrated endorectal PPA coil is better for evaluation of local prostatic cancer than is the PPA coil.

Humans↗
PubMed1993

Role of dendritic cells in the priming of CD4+ T lymphocytes to peptide antigen in vivo.

The contribution of dendritic cells (DC) in the initial priming of CD4+ T lymphocytes in vivo was examined. To assess the capacity of different APC to prime CD4+ T cells, a series of MHC class II I-E transgenic mice that differentially express I-E on APC were utilized. Transgenic mice that express I-E primarily on DC, on macrophages, and on B cells were primed with an I-E restricted peptide in vivo, and the extent of priming was determined by restimulation of CD4+ T cells in vitro with the same Ag. The results indicate that DC are required for priming of CD4+ T cells, and that the extent of priming correlates with the frequency of I-E+ DC. Moreover, DC alone can serve as APC for the peptide Ag, in that priming can be restored in an I-E negative mouse by the transfer of peptide-pulsed I-E+ DC. The potency of DC, compared with B cells and macrophages, to prime naive CD4+ T cells was confirmed with in vitro studies.

Animals↗
PubMed1992

Recycling and phosphorylation of eukaryotic initiation factor 2 on 60S subunits of 80S initiation complexes and polysomes.

Phosphorylation of the alpha-subunit (38 kDa) of eukaryotic initiation factor 2 (eIF-2 alpha) regulates initiation of protein synthesis in eukaryotic cells. This phosphorylation is enhanced in cycloheximide-treated heme-deficient reticulocyte lysates in which polysomes are maintained. In early heme deficiency prior to polysome disaggregation, eIF-2(alpha P) accumulates primarily on the 60S subunits of polysomes. Further, isolated polysomes contain eIF-2 alpha that is efficiently phosphorylated in vitro by heme-regulated inhibitor (HRI). Immunoblot analysis of eIF-2 distribution in sucrose gradients of actively protein-synthesizing lysates indicates that eIF-2 is distributed at low levels throughout the polysome profiles. These findings suggest that polysome-bound eIF-2 alpha is a target of HRI under physiological conditions. The presence of eIF-2 on the 60S subunits of polysomes is incompatible with the conventional model in which eIF-2 is recycled during the joining of the 48S preinitiation complex and the 60S subunit to form the 80S initiation complex. A modified model is presented with emphasis on the translocation of eIF-2 from the 40S ribosomal subunit of the 48S preinitiation complex (eIF-2.GTP.Met-tRNA(f).40S.mRNA) to the 60S subunit of the 80S initiation complex.

Animals↗
PubMed1990

Synthesis of the membrane fusion and hemagglutinin proteins of measles virus, using a novel baculovirus vector containing the beta-galactosidase gene.

An improved baculovirus expression vector was developed to expedite screening and facilitate oligonucleotide-directed mutagenesis. This vector contained twin promoters derived from the P10 and polyhedrin genes of Autographica californica nuclear polyhedrosis virus. The P10 promoter directed the synthesis of beta-galactosidase, whereas the polyhedrin promoter controlled the synthesis of foreign gene products. These two genes recombined with wild-type virus genome to yield recombinants which were polyhedrin negative, produced the foreign gene product, and formed blue plaques when beta-galactosidase indicator was present in the agarose overlay. An origin of replication derived from M13 or f1 bacteriophage was also included in the plasmid to permit the synthesis of single-stranded DNA. This template DNA was used to introduce or delete sequences through the process of site-specific mutagenesis. The measles virus virion possesses a membrane envelope which contains two glycoproteins: the hemagglutinin (H) and membrane fusion (F) proteins. The H polypeptide has receptor-binding and hemagglutinating activity, whereas the F protein mediates virus penetration of the host cell, formation of syncytia, and hemolysis of erythrocytes. Genes for these two glycoproteins were inserted into the NheI cloning site of the modified expression vector described above. The vector and purified wild-type viral DNA were introduced into Sf9 insect cells by calcium phosphate precipitation. A mixture of wild-type and recombinant virus was generated and used to infect Sf9 cells, which were subsequently overlaid with agarose. After 3 days, 0.1 to 1% of the plaques became blue in the presence of beta-galactosidase indicator. At least 70% of these blue viral colonies contained the foreign gene of interest as determined by dot blot analysis. Recombinant virus was separated from contaminating wild-type virus through several rounds of plaque purification. Insect cells were then infected with the purified recombinants, and synthesis of H and F proteins were verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblot detection and Coomassie blue staining. Glycosylation of the proteins appeared to be impaired somewhat, and the precursor to the F protein was not completely cleaved by the proteases present in insect host cells. On the other hand, both proteins appeared to be active in hemagglutination, hemolysis, and cell fusion assays. Levels of synthesis were in the order of 50 to 150 mg of protein per 10(8) cells.

Animals↗
PubMed1990

Persistent hypoxemia and excessive daytime sleepiness in chronic obstructive pulmonary disease (COPD).

Fourteen patients with chronic obstructive pulmonary disease (COPD) and chronic hypoxemia were studied to evaluate the relationship between hypoxemia and objective and subjective daytime sleepiness. Patients were selected with a waking PaO2 of less than 70 mm Hg and less than 50 percent predicted FEV1. Clinically, none of these patients had complaints of significant daytime sleepiness. Each patient underwent standard all-night polysomnographic evaluation followed by a multiple sleep latency test (MSLT). There was no significant correlation between the mean sleep onset latency for the MSLT and the waking PaO2, PCO2, FEV1, or spontaneous desaturations during sleep. Patients with COPD exhibited a mean MSLT that was within normal limits, despite a short total sleep time and numerous arousals from sleep. We conclude that there appears to be no relationship between chronic daytime hypoxemia and subjective reports and objective measures of daytime sleepiness.

Aged↗
PubMed1989

Antigen presentation by neonatal murine spleen cells.

The ability of murine neonatal spleen cells to present soluble antigen to T-helper cells and to produce growth factors in response to subsequent cellular interactions was studied. The T-helper-cell line (D10-G4.1) (D10), which is specific for the soluble antigen conalbumin presented on H-2-matched (H-2k) antigen-presenting cells, was used as cooperating and indicator cells in these cellular interactions. The D10 cells are TH2 T-helper cells which secrete the autocrine growth factor IL-4 and can also respond to exogenous IL-2 (T. R. Mosmann and R. L. Coffmann, Immunol. Today 8, 223, 1987). D10 cells require exogenous IL-1 for their proliferation and secrete, in addition to IL-4, IL-1 inducer factor and GM-CSF. The ability of neonatal spleen cells to present antigen and to stimulate D10 cells to produce IL-4 and proliferate is low. During antigen presentation there is an augmentation of IL-1 and IL-2 production by the antigen-presenting spleen cell population. However, neonatal spleen cells do not respond to the same levels as do adult spleen cells. The addition of exogenous IL-1 cannot repair the antigen presentation by neonatal cells. Experiments in which the antigen processing and presentation steps were separated from those requiring growth factor induction and secretion demonstrate that neonatal spleen cells are impaired in their ability to perform adequate antigen processing and presentation. The neonatal spleen cells are as competent as adult cells to cooperate with T-helper cells and secrete growth factors, provided antigen processing and presentation is performed by fully competent adult spleen cells. Experiments in which neonatal and adult antigen-presenting spleen cell populations were mixed, and others in which plastic adherent and nonadherent cells were separated, could not detect any suppressor mechanisms responsible for the low antigen presentation of neonatal cells. Thus, neonatal spleen cells are impaired in the initial stages of antigen processing and presentation. This impairment which leads to low levels of growth factor production is the major determinant in the ineffectual stimulation of T-helper cells by neonatal spleen cells.

Animals↗
PubMed1989

AIDS-related secular trends in cancer in Los Angeles County men: a comparison by marital status.

Data from the population-based cancer registry for Los Angeles County, an area with high risk of AIDS, were used to evaluate secular trends of Kaposi's sarcoma (KS), non-Hodgkin's lymphoma, and other possibly AIDS-related cancers in men aged 18 to 54. Marital status was used as a surrogate for homosexual behavior to compare the proportional incidence rates for the pre-AIDS era, 1972 to 1979, to those for 1980 to 1982 and 1983 to 1985. Both absolute incidence and proportional incidence of KS continue to increase sharply, although in absolute numbers, KS is making a smaller contribution to the total number of AIDS cases as the Los Angeles County epidemic progresses. For never-married men the proportional incidence rate of KS in 1983 to 1985 was nearly 100-fold greater than that of 1972 to 1979 and 7-fold greater than that of 1980 to 1982. High-grade lymphomas show statistically significant secular increases in both never-married and ever-married men, but only the rates of Burkitt's lymphomas have increased to a greater extent in never-married men. A small but significant increase of central nervous system lymphomas is seen in both marital status groups. There is no evidence of any AIDS-related increases in Hodgkin's disease, leukemia, testicular cancer, anal cancer, liver cancer, oral cancer, multiple myeloma, or malignant melanoma. As of 1985, cancer, as a manifestation of AIDS, is still apparently limited to KS and high-grade lymphomas (particularly Burkitt's) in Los Angeles County.

Acquired Immunodeficiency Syndrome↗