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Biomedical subjects

S N Sarkar

Publications and source records attributed to S N Sarkar.

35 records · Page 2Linked to original sources
PubMed1997

Effect of isoproturon on male reproductive system: clinical, histological and histoenzyonological studies in rats.

Isoproturon, a nonhalogenated substituted phenylurea herbicide, was evaluated for its cumulative toxic effects on testicular histomorphology., steroid hormone biosynthesis-related enzymes, spermatogenesis and sperm cells in adult albino rats. The compound, suspended in refined groundnut oil, was administered (po) to rats for 10 weeks @ 0,200, 400 and 800 mg/kg/day for 6 days/week. Isoproturon, at 800 mg/kg dose, decreased epididymal sperm count and percentage of motile sperms and increased the percentage of morphologically abnormal sperm cells. At the same dose, diameter of seminiferous tubules was reduced, number of tubules per microscopic field was increased and the percentage of tubules with evidence of spermatogenesis decreased. However, the percentage of damaged tubules was increased with 400 and 800 mg/kg doses. Histoenzymological observations revealed dose-related reduction in the activities of glucose-6-phosphate dehydrogenase and delta 5-3 beta-hydroxy steroid dehydrogenase. Activity of 17 beta-hydroxy steroid dehydrogenase was not affected appreciably. Overall findings suggest that isoproturon, at high dose, impairs androgen biosynthetic process, affects spermatogenesis and induces maturational anomalies of sperm cells in rat.

Administration, Oral↗
PubMed1996

Reversible unfolding of Escherichia coli alkaline phosphatase: active site can be reconstituted by a number of pathways.

Acid-induced and guanidine hydrochloride (GdnCl)-induced reversible unfolding of Escherichia coli alkaline phosphatase (AP) was characterized under equilibrium conditions. The protein was exposed to extreme conditions of pH 2.0 or 6 M GdnCl and was subsequently returned to normal conditions. Associated changes in the protein structure was probed by various spectroscopic methods. The changes in the functional properties were monitored by measuring enzymatic activity, capacity to renature spontaneously upon removal of the denaturant, and renaturation in presence of various site-specific and nonspecific effector molecules, in the absence and presence of beta-mercaptoethanol. Analysis of the fluorescence and CD spectra showed that the unfolding of the organized structures was much more extensive in 6 M GdnCl than at pH 2.0. Intrachain S-S bonds in each unfolded state were accessible to reduction by beta-mercaptoethanol. The effectors Zn2+ and ATP induced renaturation of active site only under reducing conditions, whereas Triton X-100 or alpha-crystallin needed the presence of some organized structure. The reconstituted protein from each denatured state without or with an effector showed different CD spectra. It is concluded that the active site domain of AP could be reconstituted independently of other structural domains in different pathways.

Alkaline Phosphatase↗
PubMed1995

Kinetic characterization of rat brain type IIA sodium channel alpha-subunit stably expressed in a somatic cell line.

1. The rat brain type IIA Na+ channel alpha-subunit was stably expressed in Chinese hamster ovary (CHO) cells. Current through the expressed Na+ channels was studied using the whole-cell configuration of the patch clamp technique. The transient Na+ current was sensitive to TTX and showed a bell-shaped peak current vs. membrane potential relation. 2. Na+ current inactivation was better described by the sum of two exponentials in the potential range -30 to + 40 mV, with a dominating fast component and a small slower component. 3. The steady-state inactivation, h infinity, was related to potential by a Boltzmann distribution, underlying three states of the inactivation gate. 4. Recovery of the channels from inactivation at different potentials in the range -70 to -120 mV were characterized by an initial delay which decreased with hyperpolarization. The time course was well fitted by the sum of two exponentials. In this case the slower exponential was the major component, and both time constants decreased with hyperpolarization. 5. For a working description of the Na+ channel inactivation in this preparation, with a minimal deviation from the Hodgkin-Huxley model, a three-state scheme of the form O<-->I1<-->I2 was proposed, replacing the original two-state scheme of the Hodgkin-Huxley model, and the rate constants are reported. 6. The instantaneous current-voltage relationship showed marked deviation from linearity and was satisfactorily fitted by the constant-field equation. 7. The time course of activation was described by an m chi model. However, the best-fitted value of chi varied with the membrane potential and had a mean value of 2. 8. Effective gating charge was determined to be 4.7e from the slope of the activation plot, plotted on a logarithmic scale. 9. The rate constants of activation, alpha m and beta m, were determined. Their functional dependence on the membrane potential was investigated.

Animals↗
PubMed1995

Subacute toxicity of urea herbicide, isoproturon, in male rats.

Isoproturon, a substituted phenylurea herbicide, was evaluated for its cumulative toxic effect on growth, organ weight and histomorphology of different organs in adult male rats. The compound (200, 400 and 800 mg/kg/day for 6 days/week), suspended in refined groundnut oil, was administered to rats p.o. for 7 and 10 weeks. There were no definite signs and symptoms of toxicity in treated rats but the herbicide significantly decreased the weekly body weight of rats at 800 mg/kg dose. Isoproturon, in all the three doses, increased the weight of liver in a dose-dependent manner. At 800 mg/kg dose, isoproturon increased the weight of kidney and heart, and decreased the weight of epididymis. Histopathological alterations in the organs were dose-dependent. Salient microscopic changes induced by isoproturon were hepatocellular degenerative changes and focal necrosis in liver, glomerular and tubular degeneration in kidney and haemosiderosis in spleen. Testis showed degeneration and desquamation of cells of germinal layers. Tubular lumens of testis and epididymis exhibited reduced number of spermatids and spermatozoa, respectively, indicating retardation of spermatogenesis.

Animals↗
PubMed1993

Effect of fenvalerate, a pyrethroid insecticide on membrane fluidity.

Fenvalerate is a commonly used pyrethroid insecticide, used to control a wide range of pests. We have studied its interaction with the membrane using fluorescence polarization and differential scanning calorimetry (DSC) techniques. Fenvalerate was found to decrease the DPH fluorescence polarization value of synaptosomal and microsomal membrane, implicating that it makes the membrane more fluid. At different concentrations of fenvalerate, the activation energy of the probe molecule in the membrane also changes revealed from the change in slope of the Arrhenius plot. At higher concentrations the insecticide slowly saturates the membrane. The effects of fenvalerate on model membrane were also studied with liposomes reconstituted with dipalmitoylphosphatidylcholine (DPPC). Fenvalerate decreased the phase transition temperature (Tm) of DPPC by 1.5 C degrees at 40 microM concentration, but there was no effect on the cooperativity of the transition as interpreted from the DSC thermogram. From the change in the thermogram profile with fenvalerate it has been interpreted that it localizes in the acyl chain region of the lipid, possibly between C10 and C16 region and weakens the acyl chain packing. Fenvalerate was also found to interact with DPPC liposomes containing cholesterol to fluidize it.

Animals↗
PubMed1993

Production of IL-3 by non-transformed primary neonatal murine keratinocytes: evidence for constitutive IL-3 gene expression in neonatal epidermis.

Interleukin 3 (IL-3) is a cytokine produced by activated T lymphocytes that is best understood as a hematopoietic growth and differentiation factor. Production of IL-3 by other cell types is controversial; while certain transformed non-lymphocyte cell lines can produce IL-3, it is generally assumed that their non-transformed counterparts do not. It has been previously reported that Pam 212, a transformed murine keratinocyte cell line, produces IL-3. In this study we report that IL-3 can also be secreted by normal murine keratinocytes. Using a cell line (FL5.12) which is responsive to IL-3 and not to other keratinocyte derived cytokines, (e.g. GM-CSF, IL-1 and IL-6), we tested conditioned media from cultures of normal neonatal keratinocytes for biologically active IL-3. These media stimulated the proliferation of FL5.12, and the effect could be neutralized by specific antibodies to IL-3. The presence of IL-3 mRNA was demonstrated by polymerase chain reaction (PCR) amplification of reverse transcribed IL-3 mRNA from cultured normal neonatal keratinocytes and confirmed by Southern blot analysis. By similar techniques, IL-3 mRNA could be identified in freshly isolated neonatal epidermis but not dermis. These data indicated that IL-3 is produced by keratinocytes in the skin of normal neonatal mice, raising the likelihood that the neonatal epidermal microenvironment may have hematopoietic or lymphopoietic properties.

Animals↗
PubMed1993

Neurotoxicity of isoproturon, a substituted phenylurea herbicide, in mice.

Effects of single oral administration of isoproturon (0.5, 1.0 and 2.0 g/kg) on CNS of male mice were studied. At higher doses, spontaneous motor activity (SMA) and forced locomotor activity (FLA) were reduced. Reduction of SMA and FLA was lesser than the reference drug-chlorpromazine hydrochloride (Ch. HCl; 15 mg/kg). Isoproturon, at all doses, potentiated both pentobarbital and barbital-induced sleeping time. At higher doses, potentiation of pentobarbital-induced hypnosis was comparable to Ch.HCl but isoproturon was more effective in inducing hypnosis with barbital. Isoproturon could not protect mice against amphetamine-induced aggregation toxicity. Isoproturon exhibited anticonvulsant activity against both supramaximal electroshock seizures and pentylenetetrazol-induced convulsions. It had no anticonvulsant activity against strychnine but only caused delay in onset of and protection from mortality. At higher doses, anticonvulsant action of isoproturon was comparable to diphenylhydantoin (50 mg/kg) and phenobarbital sodium (100 mg/kg). The study reveals that isoproturon has distinct inhibitory effect on central motor performance and sedative action on CNS. And also it has anticonvulsive and protective actions.

Amphetamine↗
PubMed1993

Fetotoxic and teratogenic potential of substituted phenylurea herbicide, isoproturon, in rats.

Effect of isoproturon (0.225, 0.45 and 0.90 g/kg/day) administered (po) from day 6 through 15 of gestation was studied on pregnant rats and their offsprings. There were no distinct clinical signs other than dose-related depression and drowsiness of pregnant rats. None of the animals died due to toxicity of isoproturon. At higher doses, decreased maternal body weight was observed during the advanced stage of pregnancy. The litter size, fetal weight and crown-rump and transumbilical lengths were decreased. There was increase in fetal resorption frequency and the number of fetuses with stunted growth. The compound had no effect on fetal sex ratio. No major visceral and skeletal malformations were observed. The study indicates fetotoxic potential of the compound.

Abnormalities, Drug-Induced↗
PubMed1987

DNA mismatch repair detected in human cell extracts.

A system to study mismatch repair in vitro in HeLa cell extracts was developed. Preformed heteroduplex plasmid DNA containing two single base pair mismatches within the SupF gene of Escherichia coli was used as a substrate in a mismatch repair assay. Repair of one or both of the mismatches to the wild-type sequence was measured by transformation of a lac(Am) E. coli strain in which the presence of an active supF gene could be scored. The E. coli strain used was constructed to carry mutations in genes associated with mismatch repair and recombination (mutH, mutU, and recA) so that the processing of the heteroduplex DNA by the bacterium was minimal. Extract reactions were carried out by the incubation of the heteroduplex plasmid DNA in the HeLa cell extracts to which ATP, creatine phosphate, creatine kinase, deoxynucleotides, and a magnesium-containing buffer were added. Under these conditions about 1% of the mismatches were repaired. In the absence of added energy sources or deoxynucleotides, the activity in the extracts was significantly reduced. The addition of either aphidicolin or dideoxynucleotides reduced the mismatch repair activity, but only aphidicolin was effective in blocking DNA polymerization in the extracts. It is concluded that mismatch repair in these extracts is an energy-requiring process that is dependent on an adequate deoxynucleotide concentration. The results also indicate that the process is associated with some type of DNA polymerization, but the different effects of aphidicolin and dideoxynucleotides suggest that the mismatch repair activity in the extracts cannot simply be accounted for by random nick-translation activity alone.

Bacterial Proteins↗
PubMed1986

Detection and analysis of UV-induced mutations in mammalian cell DNA using a lambda phage shuttle vector.

In order to study mutagenesis in mammalian cells, stable mouse L-cell lines were established with multiple copies of a lambda phage vector that contains the supF gene of Escherichia coli as a target for mutagenesis. Rescue of viable phage from high molecular weight mouse cell DNA using lambda in vitro packaging extracts was efficient (5 phage per microgram of cell DNA per copy) and yielded a negligible background of mutant phage (0 out of 54,605). From mouse cells exposed to 254-nm ultraviolet light (12J/m2), 78,510 phage were rescued, of which 8 were found to have mutant supF genes. DNA sequence analysis of the mutants suggests that the primary site of UV mutagenesis in mammalian cells is at pyrimidine-cytosine (Y-C) sequences, and that the most frequent mutation at this site is a C----T transition.

Animals↗
PubMed1986

Induction of c-myc expression in human B lymphocytes by B-cell growth factor and anti-immunoglobulin.

Purified human B lymphocytes were examined for transcriptional expression of c-myc in response to mitogenic stimulation by the method of in situ hybridization using 35S-labeled DNA probes. The level of c-myc expression increased 10- to 20-fold within 2 hr after the addition of anti-mu, formalinized Staphylococcus aureus Cowan strain I, or B-cell growth factor, as compared to resting B cells. After 72-96 hr of mitogenic stimulation, c-myc expression remained elevated 5-fold, but expression among individual cells had become more heterogeneous than at early time points. To determine whether c-myc expression in human B lymphocytes is phase specific within the cell cycle, mitogen-stimulated cells were sorted by DNA content into populations of cells in G0/G1, S, and G2/M phases of the cell cycle. Examination of c-myc expression in phase-specific cells revealed that c-myc expression was elevated in all phases of the cell cycle, but it appeared to be maximally expressed in S phase. These studies suggest that c-myc expression in normal human B lymphocytes is cell-cycle dependent and remains elevated in all phases of the cycling cell.

Antibodies, Anti-Idiotypic↗
PubMed1985

Direct and inducible mutagenesis in mammalian cells.

The understanding of mutagenic mechanisms in mammalian cells is based on extrapolations of results obtained in prokaryotes and simple eukaryotes. The use of animal viruses as targets for mutagenesis suggests that these extrapolations may be valid: recent data indicate that mutagenesis of ultraviolet-damaged templates in mammalian cells seems to occur at similar sites to that observed in prokaryotes; furthermore, nuclear replicating animal viruses are subject to the phenomenon of ultraviolet-enhanced reactivation, called SOS reactivation in bacteria. In experiments analogous to Weigle mutagenesis of phage, several groups have shown that animal cells appear to respond to DNA damage by induction of mutagenic pathways which can act upon infecting viral genomes. Recent experiments with shuttle vectors that can replicate both in animal cells and in bacteria have confirmed these conclusions. These vectors now make possible the rapid recovery of mutant genes for direct sequence analysis.

Animals↗
PubMed1980

Kinetics of induction and decay of error-prone DNA repair activity in Escherichia coli after treatment with nalidixic acid.

Inducible error-prone DNA repair activity was detected by infecting nalidixic acid-pretreated E. coli cells with UV-irradiated phage phi X174. Induction and decay kinetics of reactivation very much resembled that of mutagenesis of the UV-damaged phage. Repair as well as mutagenic activity increased for about 30 min. The maximal error-prone repair capacity, which was induced in the cell during the 30 min nalidixic acid treatment, rapidly died out during subsequent cell growth in absence of nalidixic acid. Induction of this repair mode was not observed in a recA- mutant. In the presence of nalidixic acid plus rifampicin both repair and mutagenic effects were abolished.

Bacteriophage phi X 174↗