Establishment and characterization of endometrial undifferentiated carcinoma cell line (TMCC-2.U)

We established new cell line designed TMCC-2.U, which suggested transformation to undifferentiated carcinoma, derived from endometrial clear cell carcinoma cell line (TMCC-2). The monolayer culture cell showed a pavement arrangement and spindle like shape. A rough-endoplasmic reticulum, mitochondria etc. are well developed. But cytoplasmic endocrine granulosa were so poorly, it suggests functional developments are poor. The TMCC-2.U cells were transplanted to nude mice which showed no typical pattern suggested undifferentiated carcinoma. Their chromosome number varied and the mode is 78. Marker chromosome were found frequency. Growth pattern and production of tumor marker are clearly differentiate from TMCC-2. As mensioned above, TMCC-2.U cell line will be very valuable in basic research on mechanism of transformation and effects of patient's serum on hystogenesity.     

Full replacement of 2-mercaptoethanol by cysteine plus selenium compounds in augmenting DNA synthesis of mitogen-stimulated mouse spleen lymphocytes

Mouse spleen lymphocytes require 2-mercaptoethanol for maximal mitogenic activation in vitro. Previous studies indicate that the lymphocytes are defective in the cystine transport activity and that they require 2-mercaptoethanol to utilize cystine. 2-Mercaptoethanol catalytically carries cysteine moiety into the cells in a mixed disulfide form. Because cysteine is easily oxidized to cysteine in the culture medium, it has been not easy to precisely examine the effect of near-physiological concentrations of cysteine on the activation of lymphocytes. By controlling the cysteine content in the medium, we have reviewed the effect of cysteine to see if cysteine replaces 2-mercaptoethanol in enhancing the DNA synthesis of lipopolysaccharide-stimulated lymphocytes. It was found that cysteine was less effective than 2-mercaptoethanol, and that cysteine fully replaced 2-mercaptoethanol when a selenium compound was supplemented. The effects of cysteine and selenium compounds were apparently independent and additive. Among the selenium compounds examined, sodium selenite and L-selenocystine were much more effective in stimulating DNA synthesis than sodium selenate and L-selenomethionine.     

Bullous pemphigoid antigen (BPAG1): cDNA cloning and mapping of the gene to the short arm of human chromosome 6

Bullous pemphigoid antigen (BPAG1), an integral component of the cutaneous basement membrane zone, serves as autoantigen in a blistering disease, bullous pemphigoid. In this study, we have generated cDNAs corresponding to human BPAG1 sequences. Two cDNAs, a 0.45-kb PCR product synthesized with human keratinocyte RNA as template and a 2.2-kb cDNA isolated from human keratinocyte lambda gt11 library, were utilized for chromosomal in situ hybridizations to establish the genomic location of the BPAG1 gene. Metaphase chromosomes of phytohemagglutinin-stimulated human peripheral blood leukocytes were examined by hybridizations with 3H-labeled cDNAs, and the chromosomes were identified by R-banding (fluorochrome-photolysis-Giemsa method). The results indicated that the human BPAG1 gene is at locus 6p11-6p12. This conclusion was supported by hybridizations with a panel of human X rodent hybrid cell DNA, which indicated concordance with human chromosome 6. Because different genes encoding human basement membrane components have been mapped previously to chromosomes other than 6, the results further indicate that human basement membrane zone genes are widely dispersed within the human genome.     

Genetic Control of Immune Responses to HIV-1 env DNA Vaccine

We investigated the genetic control of immunoglobulin production and the delayed-type hypersensitivity (DTH) response produced by an HIV-specific DNA vaccine using several strains of mice. Murine antigen-specific immunoglobulin production was determined by ELISA. The DTH response was assessed in terms of the footpad swelling reaction. All strains of mice, except for B10.RIII and B10.T(6R), exhibited strong immunoglobulin production and footpad swelling in response to the DNA vaccine. In vitro treatment of lymphoid cells with monoclonal antibodies showed that the footpad swelling response was mediated by CD4⁺8^? and Ia— T cells. However, CD8⁺ T cells did not suppress footpad swelling. There was no difference in the induction of HIV-specific immunoglobulin production or DTH response induced by the DNA vaccine among the strains, suggesting that HIV-specific DNA vaccine is useful for immunizing various populations against HIV-1.     

Plasma levels of glibenclamide in diabetic patients during its routine clinical administration determined by a specific radioimmunoassay

Plasma concentration of glibenclamide in routine clinical practice was determined by a specific radioimmunoassay. In diabetic patients treated with glibenclamide for a month or longer, the drug level in fasting morning plasma was variable but the mean level paralleled the daily dose. After oral administration of 2.5 mg in healthy and diabetic subjects, the drug level reached peaks in 1.5 hours and declined to the half of the peak level in next 2-3 hours. The plasma glibenclamide profile after oral dose did not differ significantly in patients with secondary failure to the drug. Comparison of a single-dose and divided-dose schedules of 5 mg glibenclamide revealed that plasma drug level increased each time after administration. Plasma glucose and insulin concentrations did not differ significantly at most times of the day but there was a tendency that increment of plasma glucose after meal was suppressed by a dose taken immediately before a meal. The relationship of blood level of glibenclamide to clinical effectiveness may be rather indirect and needs to be elucidated.