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Biomedical subjects

R Prasad

Publications and source records attributed to R Prasad.

At least 181 records · Page 10Linked to original sources
PubMed1994

Copper levels & reactive oxygen species in uterine flushings in copper intrauterine contraceptive device users.

Copper levels were estimated in the uterine flushings in 15 copper 'T' 200 device acceptors. There was a statistically significant difference in the preinsertion and one week post insertion values (P < 0.001). This also corresponded to a high level of generation of reactive oxygen species. The levels of reactive oxygen species were similar to preinsertion values at 4 and 12 wk. The copper values declined over the 3 months but were still higher at 12 wk as compared to preinsertion levels and this difference was statistically significant. Hence addition of copper to the intrauterine device besides enhancing the contraceptive efficacy, perhaps also plays an important role in microbicidal activity and helps in overcoming the infection introduced at the time of insertion.

Adult↗
PubMed1993

Cloning of the ALL-1 fusion partner, the AF-6 gene, involved in acute myeloid leukemias with the t(6;11) chromosome translocation.

Reciprocal chromosome translocations involving 11q23 are frequently associated with acute leukemias, with the t(4;11) translocation predominating among acute lymphoblastic leukemias, and the t(9;11), t(11;19) and t(6;11) translocations most common among acute myeloid leukemias. In each of these translocations the ALL-1 gene, located at 11q23 and constituting the human homologue of Drosophila trithorax, fuses to a specific gene on the partner chromosome to produce a chimeric protein. Here we report the cloning and the characterization of the partner gene from chromosome 6 (AF-6). AF-6 is expressed in a variety of cell types and encodes a protein of 1612 amino acids. The protein contains short stretches rich in prolines, charged amino acids, serines, or glutamines. In addition, the AF-6 protein contains the GLGF motif shared with several proteins of vertebrates and invertebrates thought to be involved in signal transduction at special cell-cell junctions.

Amino Acid Sequence↗
PubMed1993

Yeast open reading frame YCR14C encodes a DNA beta-polymerase-like enzyme.

We have shown by activity gel that overexpression in E. coli of a yeast chromosome 3 open reading frame (ORF) designated YCR14C and bearing homology to mammalian DNA polymerases beta results in a new DNA polymerase in the host cells. The molecular mass of this enzyme corresponded to the YCR14C-predicted 67 kDa protein, and NH2-terminal amino acid sequencing confirmed that the expressed protein was encoded by the yeast ORF. This new yeast DNA polymerase was purified to homogeneity from E.coli. In a fashion similar to that of mammalian beta-polymerases, the purified yeast enzyme exhibited distributive DNA synthesis on DNA substrate with a single-stranded template and processive gap-filling synthesis on a short-gapped DNA substrate. Activity of this yeast beta-polymerase-like enzyme was sensitive to the beta-polymerase inhibitor ddNTP and resistant to both 1 mM NEM and neutralizing antibody to E. coli DNA polymerase I. These results, therefore, indicate that YCR14C encodes a DNA beta-polymerase-like enzyme in yeast, and we name it DNA polymerase IV. Yeast strains harboring a deletion mutation of the pol IV gene are viable, they exhibit no increase in sensitivity to ultraviolet light, ionizing radiation or alkylating agents, and sporulation and spore viability are not affected in the mutant.

Amino Acid Sequence↗
PubMed1993

Effect of miconazole on the structure and function of plasma membrane of Candida albicans.

The primary mode of action of azoles is the inhibition of cytochrome P-450 dependent 14 alpha-demethylase, a key enzyme in ergosterol biosynthesis in fungi. Our results demonstrated that Candida albicans cells grown in the presence of 10 micrograms ml-1 of miconazole (miconazole-grown), do not possess ergosterol in their plasma membranes and this ergosterol depletion leads to a drastic change in membrane fluidity as shown by fluorescence polarization measurements and unsaturation index. There was an increase in membrane order in miconazole-grown cells and a reduced rate of uptake of amino acids. We also checked for membrane permeability changes in normal mid-log phase cells (normal-grown) in short incubations (10 min) with 10 micrograms ml-1 miconazole (miconazole-incubated). Interestingly, the amino acid uptake rates except that of Gly were not affected significantly in these cells. The results suggest that in the miconazole-incubated cells, the drug is not able to alter the level of ergosterol or inhibit ergosterol biosynthesis during 10 min incubation and therefore the interaction of the drug neither leads to significant disorganization of membrane components, nor affects permease activity, whereas in the miconazole-grown cells there is ergosterol depletion leading to accumulation of biosynthetic intermediates, resulting in membrane rearrangement thereby causing a major fluidity change. This fluidity change may explain the drastic reduction of amino acid transport in miconazole-grown cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1993

Identification of residues in the single-stranded DNA-binding site of the 8-kDa domain of rat DNA polymerase beta by UV cross-linking.

Rat DNA polymerase beta (beta-pol) is a 39-kDa monomeric protein, organized in two structurally and functionally distinct domains. The 8-kDa NH2-terminal domain binds single-stranded (ss) DNA, whereas the 31-kDa COOH-terminal domain does not. To facilitate studies on ssDNA binding structure-function relationships of beta-pol, we overexpressed the 8-kDa domain in Escherichia coli, and purified the recombinant protein to homogeneity. Single-stranded nucleic acid binding of the recombinant 8-kDa domain was found to be similar to that previously reported for the 8-kDa fragment prepared by proteolysis of intact beta-pol (Kumar, A., Widen, S. G., Williams, K. R., Kedar, P. Karpel, R. L., and Wilson, S. H. (1990b) J. Biol. Chem. 265, 2124-2131; Casas-Finet, J. R., Kumar, A., Morris, G., Wilson, S. H., and Karpel, R. L. (1991) J. Biol. Chem. 266, 19618-19625). Residues in or near the DNA-binding pocket of the recombinant 8-kDa domain were examined by photochemical cross-linking to [32P] p(dT)16. Cross-linking was localized to a tryptic fragment spanning residues 28 through 35 and a V8 protease fragment spanning residues 27 through 58. Sequence analysis of the various [32P]p(dT)16-labeled proteins indicated that Ser30 and His34 were modified by cross-linking to p(dT)16. Therefore, these residues of the ssDNA-binding domain of beta-pol appear to be in close contact with this nucleic acid probe.

Amino Acid Sequence↗
PubMed1993

Location of the enterotoxin gene from Salmonella typhimurium and characterization of the gene products.

The enterotoxin gene (stn) in Salmonella typhimurium (Q1 strain) was confined to an 800 bp ClaI-EcoRI genomic DNA fragment (pCE3) that coded for two polypeptides (25 and 12 kDa) under the control of the T7 RNA polymerase/promoter system. The appearance of the 25 kDa protein corresponded to the enterotoxic activity, as determined by elongation of Chinese hamster ovary (CHO) cells, fluid accumulation in rabbit intestinal loops, and altered vascular permeability in rabbit skin. The stn gene products (STN) caused an elevation of intracellular cAMP in CHO cells. These values were at control levels in stn mutants devoid of enterotoxicity, and the 25-kDa protein concurrently disappeared. The biological activity of the heat-labile enterotoxin was blocked by GM1 ganglioside and neutralized by affinity-purified antibodies made against cholera toxin. The 12 kDa protein however was not correlated with an enterotoxic response.

Animals↗
PubMed1993

Genes on chromosomes 4, 9, and 19 involved in 11q23 abnormalities in acute leukemia share sequence homology and/or common motifs.

Chromosome translocations involving band 11q23 are associated with human acute leukemias. These translocations fuse the ALL-1 gene, homolog of Drosophila trithorax and located at chromosome band 11q23, to genes from a variety of chromosomes. We cloned and sequenced cDNAs derived from transcripts of the AF-4 and AF-9 genes involved in the most common chromosome abnormalities, t(4:11)(q21:q23) and t(9:11)(p22:q23), respectively. Sequence analysis indicates high homology between the AF-9 gene protein product and the protein encoded by the ENL gene fused to ALL-1 in (11:19) chromosome translocations. AF-4, AF-9, and ENL proteins contain nuclear targeting sequences as well as serine-rich and proline-rich regions. Stretches abundant in basic amino acids are also present in the three proteins. These results suggest that the different proteins fused to ALL-1 polypeptide(s) provide similar functional domains.

Acute Disease↗
PubMed1993

Levels of plasma membrane H(+)-ATPase do not change during growth and morphogenesis of Candida albicans.

A transient rise in the PM-ATPase activity was observed at the time of commitment of Candida albicans cells to either bud or hyphal formation. However, the changes in PM-ATPase activity did not correlate with the level of enzyme protein detected by ELISA. It was found to be fairly constant during differentiation, implying that there was no de novo synthesis of the protein. Post-translational modification(s) of enzyme protein is suggested to account for variation in PM-ATPase activity during morphogenesis.

Candida albicans↗
PubMed1993

1-Aryl-2-amino/hydrazino-4-phenyl-1,6-dihydro-1,3,5-triazine-6-thione and related thiocarbamides/thiosemicarbazides as antithyroidal agents.

Different 1-aryl-2-benzylmercapto-4-phenyl-1,6-dihydro-1,3,5-triazine-6- thiones have been synthesized by known methods. These triazines on treatment with ammonia/hydrazine hydrate afforded the corresponding 1-aryl-2-amino/hydrazino-4-phenyl-1,6-dihydro-1,3,5-triazine-6-thiones which on treatment with arylisothiocyanates afforded the related thiocarbamide/thiosemicarbazides. Some of these compounds show appreciable antithyroidal activity.

Animals↗
PubMed1993

Hearing impairment and ear pathology in Nepal.

A stratified random cluster sample of 15,845 subjects was performed in two regions of Nepal to determine the prevalence and main causes of hearing impairment (the most common disability) and the prevalence of ear disease. Subjects reporting current ear pain, or ear discharge, or hearing impairment on direct questioning by a Nepali health worker (primary screening failed), had otoscopy and audiometry (using the Liverpool Field Audiometer) performed, and a questionnaire administered relating to past history. In every fifth house subjects who passed the primary screening (1,716 subjects) were examined to assess the false negative rate of screening. An estimated 16.6 per cent of the study population have hearing impairment (either ear worse than 30 dB hearing threshold level (HTL) 1.0-4.0 kHz, or 50 dB HTL 0.5 kHz), and 7.4 per cent ear drum pathology, equivalent to respectively 2.71 and 1.48 million people extrapolated to the whole of Nepal. Most hearing impairment in the school age group (55.2 per cent) is associated with otitis media or its sequelae. Probably at least 14 per cent of sensorineural deafness is preventable (7 per cent infectious disease, 3.9 per cent trauma, 0.8 per cent noise exposure, 1 per cent cretinism, and 1 per cent abnormal pregnancy or labour). Most individuals reporting current ear pathology (61 per cent) had never attended a health post, and of those receiving ear drop treatment, 84 per cent still had serious pathology. Of subjects who reported ear drop treatment at any time, 31 per cent still had serious pathology. The use of traditional remedies was prevalent.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

Capillary filtration coefficients using laser densitometry and gravimetry in isolated dog lungs.

We compared pulmonary capillary filtration coefficients (Kf,c) using measurements of transcapillary filtration rates based on laser densitometry of perfusate hematocrit changes (Jy,l) and gravimetric measurement of the rate of lobe weight gain (delta Wt/delta t) after an increase in capillary pressure (Pc) in isolated autologous blood-perfused dog lungs. Although the lobe weight increased at a rate that decreased with time, the densitometric filtration rate was relatively constant over time. Kf,c values were calculated in milliliters per minute per centimeter water per 100 g from filtration rates obtained by 1) extrapolation of the delta Wt/delta t from 3 to 10 min back to time 0 [Kf,c(0)], 2) use of the slope of the line fitted to the delta Wt/delta t and Jv,l values obtained after three stepwise increases in Pc [Kf,c(slope)], and 3) use of delta Wt/delta t and Jv,l values obtained 10, 20, and 30 min after either a constant 30-min or three 10-min stepwise increases in Pc [Kf,c(t)]. The mean Kf,c(0) values ranged from 0.171 +/- 0.024 to 0.188 +/- 0.070 and were not significantly different between vascular pressure states. These Kf,c(0) values were significantly higher than all gravimetric or densitometric Kf,c(t) or Kf,c(slope) values that ranged from 0.037 to 0.100. Kf,c(t) values were not statistically different from each other by use of either method, but the gravimetric Kf,c(slope) was significantly higher than the densitometric value. Therefore, extrapolated Kf,c(0) probably overestimates endothelial barrier Kf,c because of persistent vascular stress relaxation after a vascular pressure increase. Adjustment of tissue Starling forces may occur rapidly after an increase in filtration pressure.

Animals↗