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N Srivastava

Publications and source records attributed to N Srivastava.

At least 73 records · Page 4Linked to original sources
PubMed1995

GABA attenuates the neurotoxic effects of ethanol in neuron-enriched cultures from 8-day-old chick embryo cerebral hemispheres.

Neuron-enriched cultures were prepared from 8-day-old chick embryo cerebral hemispheres and exposed to ethanol (50 mM), GABA (10(-5) M) and ethanol (50 mM) + GABA (10(-5) M) from day 4 to 8 in culture. At day 8, control, ethanol, GABA and ethanol + GABA-treated cultures were examined morphologically and biochemically. Choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) activities were used as markers for cholinergic and GABAergic neuronal phenotypic expression, respectively. Control cultures showed more numerous and large neuronal aggregates as well as prominent neuritic bundles. Moreover, cultures treated with GABA depicted even more numerous neuronal aggregates with interconnecting neurites as compared to control. In contrast, ethanol-treated cultures exhibited smaller neuronal aggregates with less prominent neuritic bundles than control. However, cultures treated concomitantly with ethanol + GABA exhibited numerous and larger aggregates than cultures treated with ethanol alone. Neuritic bundles which were highly reduced in ethanol-treated cultures became prominent in the presence of GABA. As previously reported, ethanol alone enhanced ChAT and reduced GAD activities. GABA given alone enhanced the expression of both neuronal phenotypes. When GABA was given concomitantly with ethanol the decline in GAD and the rise in ChAT observed in ethanol-treated cultures was restored by GABA to almost control levels. Thus, ethanol-induced alterations in morphology and neuronal phenotypes were counteracted by the neurontrophic effect of GABA.

Animals↗
PubMed1995

Hypothalamic tissue stimulates hippocampal pyramidal neuron survival during development: evidence from intraocular double transplants.

The present study was undertaken to evaluate innervation and possible growth promotion by posterior hypothalamic tissue on different areas that are, or are not, interactive with this brain region during development. Posterolateral hypothalamus was dissected from embryonic day 17 rat fetuses, and inserted into the anterior chamber of the eye of adult rat hosts. Two weeks postgrafting, a second transplant consisting of either fetal hippocampal, cerebellar, or lung tissue was placed adjacent to the first graft. Growth of the intraocular double transplants was monitored weekly by measurements through the cornea. Fetal hippocampal tissue grew significantly larger when placed together with a hypothalamic graft, as compared to single hippocampal transplants. Cerebellar or lung tissue growth was not stimulated by a hypothalamic cograft. Pyramidal neuron cell counts demonstrated a significantly higher final number of these neurons in growth-stimulated hippocampal grafts, as compared to non-stimulated single hippocampal grafts. Immunohistochemistry with antibodies directed against histamine or histidine decarboxylase revealed that hippocampal transplants received the most dense histaminergic innervation. Cerebellar transplants contained occasional histaminergic neurites, and lung tissue never exhibited any histaminergic innervation from the adjacent hypothalamic graft. Taken together, these results demonstrate a growth-promoting effect of posterior hypothalamic tissue on developing hippocampal tissue, as well as target specificity of histaminergic innervation patterns.

Animals↗
PubMed1995

Astrocyte differentiation is enhanced in chick embryos treated with ethanol during early neuroembryogenesis.

In this study, we examined the effects of ethanol administered to chick embryos, on the maturation of astrocytes, using glutamine synthetase (GS) activity as an astrocyte marker. Ethanol (50 mM) was administered in ovo via the air sac, embryos were sacrificed at various days of embryonic development and GS activity was determined in cerebral hemispheres and cerebellum. We found that in both cerebral hemispheres and cerebellum, GS activity was higher in the ethanol-treated embryos, as compared to controls, during the embryonic periods, E6 to E10 in the cerebral hemispheres and E10 to E14 in the cerebellum. These periods are characterized by increased neuronal differentiation in these CNS areas. The increase in GS activity in the ethanol-treated embryos is speculated to reflect either a transient reactive gliosis and/or an enhancement in the differentiation of radial glia, immature glia, to more mature astrocytes.

Animals↗
PubMed1995

Maturation of cerebellar granule cells is delayed in cultures derived from ethanol-treated chick embryos: survival and proliferation studies.

Previous studies from this laboratory have shown that ethanol administration to chick embryos during embryonic days 1-3, a critical period of neuroembryogenesis, differentially affects primordial CNS structures. In this study, chick embryos were treated in ovo with ethanol (10 mg/50 microliter/day) at E1 to E3. At 14 days of embryonic age cerebellar (E14CE) granule cell cultures were prepared from both control and ethanol-treated embryos. Growth patterns were evaluated morphologically and the neuronal nature of these cultures was evaluated immunocytochemically. E14CE granule cell cultures exhibited neurofilament immunoreactivity demonstrating the neuronal-nature of these cultures. In addition E14CE granule cultures contained numerous glutamatergic neurons as assessed by positive glutamate immunoreactivity and also some GABAergic neurons as assessed by positive GABA immunoreactivity. Cultures derived from both control and ethanol-treated embryos were labeled with 3H-thymidine and assessed for effects on survival and proliferation in culture. Cultures derived from ethanol-treated embryos showed a higher rate of proliferation and survival during the first 3 days in culture as compared to those derived from controls. However, after 3 days in culture, survival was lower in the cultures from ethanol-treated embryos as compared to those derived from controls. We interpret these findings to mean that (a) ethanol arrested cerebellar granule cell development at an immature state; (b) immature neurons have a higher survival capacity than differentiated neurons; and (c) ethanol accelerates normal neuronal cell death as previously reported.

Animals↗
PubMed1995

Ethanol neurotoxicity on neuroblast-enriched cultures from three-day-old chick embryo is attenuated by the neuronotrophic action of GABA.

In the present study, using neuroblast-enriched cultures derived from three-day-old chick embryos (E3WE), we examined the morphological effects of ethanol and/or GABA, as well as the developmental profile of the cholinergic and GABAergic neuronal phenotypes, as assessed by the activities of choline acetyltranferase (ChAT) and glutamate decarboxylase (GAD). Cultures exposed to ethanol (50 mM) exhibited smaller and fewer aggregates than controls with a neuritic network that lacked fasciculation. In cultures treated with GABA (10(-5) M) alone or ethanol+GABA the size and number of the neuronal aggregates was increased and also neuritic arborization and fasciculation was enhanced. Thus, addition of GABA restored the normal growth pattern in the ethanol-treated cultures. As previously shown, E3WE culture treated with ethanol alone showed a decrease in both ChAT and GAD activities compared to controls. Both cholinergic and GABAergic neuronal phenotypes were enhanced in cultures treated with GABA as assessed by increases in ChAT and GAD activities, respectively, compared to controls. Moreover, in cultures treated concomitantly with ethanol and GABA both ChAT and GAD activities were higher than in ethanol-alone-treated cultures. Thus, the presence of GABA in the ethanol-treated cultures counteracted the decline in ChAT and GAD activities observed in the ethanol-alone-treated cultures. We conclude that GABA through its neuronotrophic actions can rescue neuroblasts from ethanol insult and restore neuronal phenotypes.

Animals↗
PubMed1995

Rifamycins: strain improvement program.

Rifamycins are primarily produced by Gram-positive bacterium Amycolatopsis mediterranei, which belongs to the order Actinomycetales. These antibiotics, apart from their application against pathogens of tuberculosis and leprosy, have also been found to be effective against several other pathogens including Mycobacterium avium and Pneumococcus. Because of the importance of rifamycin, the producer strain A. mediterranei has been genetically manipulated since 1957 in order to develop a strain that can either produce larger amounts of rifamycin or derivatives of rifamycin. In this article, the importance of the producer strain, traditional methods (mutations and recombination) of strain improvement, their limitations, and the development of a cloning vector and transformation methods that have made recombinant DNA techniques accessible for genetic manipulations of A mediterranei are discussed.

Actinobacteria↗
PubMed1995

A rapid method for the preparation of plasmid DNA.

A rapid method was developed to prepare plasmid DNA to be used for in vitro transcription, subcloning and sequencing. Clean plasmid DNA can be prepared within 90 min using this method and the quality of the plasmid DNA was found to be suitable for sequencing, preparation of in vitro RNA transcript and electroporation. Using this method plasmid DNA can be prepared from 2 ml to 100 ml culture. The yield of the recombinant plasmids was found to be in the range of 50-100 micrograms from 10 ml culture depending on the low and high copy number plasmids. The advantage of this method is that one can easily and rapidly prepare large quantities of plasmid DNA of high quality to be used for various purposes.

DNA↗
PubMed1995

Regulation of low density lipoprotein receptor gene expression in HepG2 and Caco2 cells by palmitate, oleate, and 25-hydroxycholesterol.

Our in vivo studies in mice have shown that LDL-receptor gene expression is regulated differently in both liver and intestine by dietary cholesterol and dietary saturated fat. While dietary cholesterol serves to regulate at transcriptional levels, dietary fatty acids do not. To study the mechanism of regulation of LDL-receptor by saturated fat and cholesterol at the cellular level, where any secondary effects of long-term feeding in vivo are minimized we used the cultured hepatoma and colon carcinoma cells, HepG2 and Caco2. LDL-receptor activity was determined by 125I-labeled LDL binding and uptake, LDL-receptor protein by Western blotting, LDL-receptor mRNA by RNase protection assay, and relative rates of LDL-receptor mRNA transcription by nuclear 'run-off' assay. Incubation of cells in lipoprotein-deficient serum (LPDS) for 48 h progressively induced LDL-receptor activity and LDL-receptor protein by 5- to 6-fold in HepG2 cells and 2- to 3-fold in Caco2 cells. Absolute levels of LDL-receptor mRNA and relative rates of LDL-receptor mRNA transcription also increased in parallel to the LDL-receptor activity and protein levels in both cell lines. These data suggest that LPDS induced the LDL-receptor gene by transcriptional mechanism. The suppressive effect of 25-hydroxycholesterol on LDL-receptor regulation was studied by incubating HepG2 and Caco2 cells grown either in 10% FCS or 10% LPDS for 24 h and then for 0-24 h with various doses of 25-hydroxycholesterol. In HepG2 cells, LDL-receptor activity and protein mass progressively decreased to 50% of zero time controls over 24 h. LDL-receptor mRNA levels and relative rates of transcription decreased in parallel. In Caco2 cells, 25-hydrocholesterol lowered LDL-receptor activity, mRNA, and transcription by approximately 35%. To examine the effects of palmitate on LDL-receptor regulation, palmitate was complexed with albumin. Palmitate decreased LDL-receptor activity by 25% in HepG2 cells without altering LDL-receptor mass, mRNA levels, or rates of mRNA transcription. Similarly, in Caco2 cells, palmitate decreased LDL-receptor activity and protein mass 30% of controls, but did not change LDL-receptor mRNA levels and/or rates of transcription. The combination of palmitate (0.8 mM) and 25-hydroxycholesterol (2.5-5 micrograms/ml) suppressed LDL-receptor activity by 65% in HepG2 cells and by 52% in Caco2 cells. However, LDL-receptor mRNA decreased by approximately 50% in HepG2 cells and 30-40% in Caco2 cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Blood↗
PubMed1995

Hypoxia-induced arterial endothelin does not influence peripheral vascular tone.

The effect of hypoxia (SpO2 80%) on the arteriovenous production of endothelin-1 (ET) in the forearm was investigated in six healthy volunteers. In eight other subjects the local vascular effects of a low physiologic dose of intra-arterially administered ET (0.5 pg/kg/min) was studied during normoxia. Forearm blood flow (FBF) was measured by venous occlusion plethysmography, and arterial and venous plasma concentrations of ET were measured. FBF was not significantly influenced during hypoxia. After 15 min of hypoxia the arterial plasma level of ET increased from 2.5 +/- 0.1 to 3.3 +/- 0.3 ng/L (p < 0.05), whereas the venous plasma level remained unchanged. Intra-arterial infusion of ET increased the arterial plasma level of ET in the forearm from 2.2 +/- 0.3 to 4.9 +/- 0.4 ng/L. This increase in ET concentration did not influence FBF. It is concluded that hypoxia is a stimulus for ET release from the pulmonary vascular bed and that the resulting increase arterial plasma ET level does not influence peripheral vascular tone.

Adult↗
PubMed1994

Outcome in relation to Apgar score in term neonates.

Sixty four asphyxiated term babies (Apgar score of 6 or less at 5 minutes) and 90 non-asphyxiated term babies (controls) were studied. Of these, 40 cases and 48 controls could be followed up. Mortality and neurodevelopmental outcome were studied in both the cases and controls. Mortality and poor neurodevelopmental outcome correlated inversely with the Apgar scores at 5 and 10 minutes. The outcome of babies with low 5 minute Apgar scores was significantly better than those with the same scores at 10 minutes. Symptomatic neonates when compared to asymptomatic neonates with same Apgar score showed significantly poorer outcome. Babies with Apgar scores of 6 at 5 or 10 minutes behaved like the controls both in terms of mortality and neurodevelopmental outcome.

Apgar Score↗
PubMed1994

Rejuvenation of epididymal principal cells by prolactin under androgen deficiency.

Influence of prolactin on the ultrastructure of principal cells lining the epididymal epithelium was investigated in Wistar rats. Orchidectomy produced degenerative changes suggesting that structural integrity of principal cell is maintained by factors originating in the testis. The atrophic changes in the principal cell of ordhidectomised rats were significantly reversed when prolactin was administered to these animals. The number of cells that responded were found to increase with the dose of prolactin injected. On the otherhand, bromocryptine treatment did not appreciably change the ultrastructure of principal cells in orchidectomised rats. Results suggest that prolactin may have a rejuvenating epididymal principal cells in androgen deficient states.

Androgens↗
PubMed1994

Impact of prolactin on epididymal lipid profile in castrated rats.

Prolactin treatment to castrated rats led to accumulation of triacylglycerol and esterified cholesterol. There was no appreciable drift in epididymal cholesterol: phospholipid ratio between the prolactin treated and control animals. However, further analysis of phospholipids showed a build up of phosphatidyl inositol, phosphatidyl choline and phosphatidyl ethanolamine but a drop in the levels of phosphatidyl serine and sphingomyelin in prolactin treated castrated rats as compared to those castrated animals injected with vehicle alone. Changes in phospholipids reported above were prominently seen in the group of castrated rats that received 100 micrograms oPRL/100 g body weight but not in those animals which received either lower or higher doses of the hormone. Interestingly, bromocryptine treatment in castrated rats produced a general depletion in the levels of all lipid classes studied in the epididymis. It is suggested that this may be due to impaired synthesis and/or increased breakdown of lipids in this organ.

Animals↗
PubMed1993

Comparative kinetic studies on aflatoxin B1 binding to pulmonary and hepatic DNA of rat and hamster receiving the carcinogen intratracheally.

Several epidemiological studies have discussed the outcome of inhalation of airborne aflatoxins by humans. Metabolism of aflatoxin B1 (AFB1) by lung parenchyma leading to DNA binding is reported here. The tissue distribution pattern of [3H]AFB1 radioactivity revealed the lungs to be the second most important organ after the liver to retain a considerable amount of the radioactivity (66%). The lung indicated a selective activation of AFB1 as it showed only 7.7% binding of [3H]AFB1 to pulmonary DNA. Rats and hamsters were dosed with [3H]AFB1 (2 microCi containing 40 micrograms AFB1/100 g body wt.) intratracheally (i.t.) and sacrificed at different intervals after toxin treatment. Peak binding occurred at 0.5, 1, and 2 h in case of hamster lung, rat lung, and alveolar macrophages of both the species, respectively. At the end of 24 h, the relative AFB1-DNA binding (percentage of peak binding) in hamster lung was 72% while that in rat was 24%. The relative binding in rat lung alveolar macrophages (AMs) was generally higher than that of the hamster. AFB1 binding to hepatic DNA of both the species approached the peak at 1 h after the toxin administration i.t. Under these conditions, binding of AFB1 (or its metabolites translocated to liver) to hepatic DNA of both the species progressively diminished with time in contrast to lung, as revealed by the relative binding values at 12 h for rat and hamster lung, which were 48 and 67%, respectively, while for the rat and hamster liver they were 28 and 24%, respectively. Binding of i.t. administered [3H]AFB1 to rat liver DNA is only marginally higher than that observed with hamster liver, in contrast to the wide difference observed in animals receiving AFB1 intraperitoneally. These results highlight the persistence of AFB1 binding to pulmonary DNA, and the extent of translocated AFB1 binding to hepatic DNA presents an interesting difference from that observed when the toxin was administered through a gastrointestinal route. It is worth concluding that AMs unlike many other xenobiotics, possess specific mixed function oxidase activity to epoxidize AFB1.

Aflatoxin B1↗
PubMed1993

Glutathione S-transferase mu polymorphism does not explain variation in nitroglycerin responsiveness.

OBJECTIVE: To determine whether the considerable interindividual variability in nitroglycerin-induced venodilation in humans is related to the polymorphic expression of the mu class of glutathione S-transferase (GST mu). Recently vascular glutathione S-transferase (EC 2.5.1.18) of the mu-class (GST mu), a polymorphic group of enzymes present in only about 60% of the population, have been identified and shown in vitro to possess high metabolic activity toward nitroglycerin. Their clinical relevance is unknown. DESIGN: Dose-response relationships to nitroglycerin were constructed in vivo measuring changes in compliance of dorsal hand veins in 26 healthy volunteers during local infusion of small amounts of nitroglycerin. Polymerase chain reaction was applied to detect the deoxyribonucleic acid sequence that codes GST mu in whole blood samples. RESULTS: The GST mu isozyme was present in 15 subjects (58%) and deficient in 11 subjects. Values for mean maximum venodilation (Emax) and dose rates producing 50% of Emax (ED50) were not significantly different between the groups with or without GST mu. The respective values were 98% and 103% dilation for Emax and 9 and 16 ng/min for ED50. There was no gender difference in the venodilatory response to nitroglycerin. CONCLUSIONS: Subjects lacking GST mu can clearly respond normally to nitroglycerin, and the large interindividual variability in nitroglycerin potency is not related to the expression of this polymorphic enzyme. Intersubject variability is therefore more likely to be the result of differences in the presence or activity of other vascular enzymes or in steps further distal in the venodilatory cascade.

Adult↗
PubMed1993

Lack of role of endothelium-derived relaxing factor in effects of alpha-adrenergic agonists in cutaneous veins in humans.

In some blood vessels, the alpha 2-adrenergic agonist clonidine simultaneously activates vasoconstrictive alpha-adrenoceptors on smooth muscle cells and endothelial alpha 2-adrenoceptors mediating release of endothelium-derived relaxing factor (EDRF), with the net vascular response representing a balance between these two counteracting pathways. To investigate whether clonidine's modest constrictor effect in human veins is due to simultaneous release of EDRF, the dorsal hand vein compliance technique was used to measure vascular responses in healthy volunteers. Clonidine-induced venoconstriction was not potentiated by methylene blue, an inhibitor of EDRF-mediated relaxation. After preconstriction with angiotensin II, clonidine did not cause venodilation but rather promoted further constriction, which could be reversed by the alpha 1-antagonist labetalol. However, in veins preconstricted with the full alpha 1-agonist phenylephrine, clonidine induced venodilation, suggesting that clonidine is a partial agonist at venous alpha 1-adrenoceptors. In conclusion, we found no evidence for endothelial alpha 2-adrenoceptor-mediated release of EDRF in human hand veins. The results further suggest that postjunctional alpha 1-adrenoceptors participate in clonidine-induced venoconstriction in humans.

Adrenergic alpha-Agonists↗
PubMed1992

Piperine, a plant alkaloid of the piper species, enhances the bioavailability of aflatoxin B1 in rat tissues.

Piperine is known to modify the biotransformation of drugs. The effect of piperine on the metabolic activation and distribution of [3H]-aflatoxin B1 (AFB1) in rats has been described. Piperine markedly inhibited liver microsome-catalysed [3H]AFB1 binding to calf thymus DNA in vitro, in a dose dependent manner. Rats pretreated with piperine accumulated considerable [3H]AFB1 radioactivity in plasma and in the tissues examined as compared to the controls. However, piperine had no influence on hepatic [3H]AFB1-DNA binding in vivo, which could possibly be due to the null effect of piperine on liver cytosolic glutathione (GSH) 5-transferase activity. Piperine-treated rat liver microsomes demonstrated a tendency to enhance [3H]AFB1 binding to calf thymus DNA in vivo. The effect of piperine on AFB1 metabolism thus closely resembles the mode of action of SKF 525-A on biotransformation of foreign compounds.

Aflatoxin B1↗
PubMed1992

Characterization of the RNA processing enzyme RNase III from wild type and overexpressing Escherichia coli cells in processing natural RNA substrates.

1. A precursor to small stable RNA, 10Sa RNA, accumulates in large amounts in a temperature sensitive RNase E mutant at non-permissive temperatures, and somewhat in an rnc (RNase III-) mutant, but not in an RNase P- mutant (rnp) or wild type E. coli cells. 2. Since p10Sa RNA was not processed by purified RNase E and III in customary assay conditions, we purified p10Sa RNA processing activity about 700-fold from wild type E. coli cells. 3. Processing of p10Sa RNA by this enzyme shows an absolute requirement for a divalent cation with a strong preference for Mn2+ over Mg2+. Other divalent cations could not replace Mn2+. 4. Monovalent cations (NH+4, Na+, K+) at a concentration of 20 mM stimulated the processing of p10Sa RNA and a temperature of 37 degrees C and pH range of 6.8-8.2 were found to be optimal. 5. The enzyme retained half of its p10Sa RNA processing activity after 30 min incubation at 50 degrees C. 6. Further characterization of this activity indicated that it is RNase III. 7. To further confirm that the p10Sa RNA processing activity is RNase III, we overexpressed the RNase III gene in an E. coli cells that lacks RNase III activity (rnc mutant) and RNase III was purified using one affinity column, agarose.poly(I).poly(C). 8. This RNase III preparation processed p10Sa RNA in a similar way as observed using the p10Sa RNA processing activity purified from wild type E. coli cells, confirming that the first step of p10Sa RNA processing is carried out by RNase III.

Chromatography, Liquid↗