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S Bhattacharya

Publications and source records attributed to S Bhattacharya.

At least 19 recordsLinked to original sources
PubMed1995

Soluble ligands of the alpha v beta 3 integrin mediate enhanced tyrosine phosphorylation of multiple proteins in adherent bovine pulmonary artery endothelial cells.

Binding of substrate-bound extracellular matrix proteins to cell surface integrins results in a variety of cellular responses including adhesion, cytoskeletal reorganization, and gene expression. We have previously shown that addition of soluble SC5b-9, the complement-vitronectin complex, resulted in an RGD-dependent increase in lung venular hydraulic conductivity (Ishikawa, S., Tsukada, H., and Bhattacharya, J. (1993) J. Clin. Invest. 91, 103-109). To identify specific integrin(s) and signal transduction pathways that are responsive to soluble vitronectin-containing ligands, we exposed confluent bovine pulmonary artery cells to purified soluble human mono- or multimeric vitronectin, or SC5b-9, and determined the extent of endothelial cell protein tyrosine phosphorylation. Monomeric vitronectin (Vn) did not induce enhanced protein tyrosine phosphorylation. However, multimeric Vn and SC5b-9 elicited time- and concentration-dependent increases in tyrosine phosphorylation of numerous proteins. Antiserum against vitronectin, RGD peptides, and monoclonal and polyclonal antibodies against the alpha v beta 3 integrin blocked the vitronectin- or SC5b-9-induced enhanced accumulation of tyrosine phosphoproteins, while antibodies against beta 1 integrins and the alpha v beta 5 integrin did not. Clustering of the alpha v beta 3 integrin using monoclonal antibody LM609 caused a pattern of enhanced tyrosine phosphorylation similar to that caused by multimeric Vn and SC5b-9, suggesting that aggregation of alpha v beta 3 was critical for signaling. Among the proteins that underwent enhanced tyrosine phosphorylation in response to vitronectin were the cytoskeletal proteins paxillin, cortactin, and ezrin, as well as the SH2 domain-containing protein Shc, and p125FAK. We conclude that ligation of the alpha v beta 3 integrin by soluble ligands promotes enhanced phosphorylation of several proteins implicated in tyrosine kinase signaling and suggest that this pathway may be important in inflammatory states which are accompanied by accumulation of SC5b-9.

Animals↗
PubMed1995

Evolutionary origins of apoB mRNA editing: catalysis by a cytidine deaminase that has acquired a novel RNA-binding motif at its active site.

The site-specific C to U editing of apolipoprotein B100 (apoB100) mRNA requires a 27 kDa protein (p27) with homology to cytidine deaminase. Here, we show that p27 is a zinc-containing deaminase, which operates catalytically like the E. coli enzyme that acts on monomeric substrate. In contrast with the bacterial enzyme that does not bind RNA, p27 interacts with its polymeric apoB mRNA substrate at AU sequences adjacent to the editing site. This interaction is necessary for editing. RNA binding is mediated through amino acid residues involved in zinc coordination, in proton shuttling, and in forming the alpha beta alpha structure that encompasses the active site. However, certain mutations that inactivate the enzyme do not affect RNA binding. Thus, RNA binding does not require a catalytically active site. The acquisition of polymeric substrate binding provides a route for the evolution of this editing enzyme from one that acts on monomeric substrates.

APOBEC-1 Deaminase↗
PubMed1995

Ras membrane targeting is essential for glucose signaling but not for viability in yeast.

Ras proteins are small GTP binding proteins that serve as critical relays in a variety of signal transduction pathways in eukaryotic cells. Like most metazoan Ras proteins, yeast Ras is post-translationally modified by addition of a farnesyl and a palmitoyl moiety, and these modifications are required for targeting the protein to the cytoplasmic face of the plasma membrane and for biological activity of the protein. We have constructed mutants of the yeast (Saccharomyces cerevisiae) Ras that are farnesylated in vivo but are not palmitoylated. These mutant proteins are not localized to the plasma membrane but function in the cell as well as the wild-type protein. Such mutants are viable but fail to induce a transient increase in intracellular cAMP concentration in response to glucose addition, although this deficiency does not yield a marked growth phenotype. These results are consistent with the hypothesis that the essential role of the farnesyl moiety on yeast Ras is to enhance productive interaction between Ras and its essential downstream target, adenylyl cyclase, rather than to localize Ras to the plasma membrane.

Alleles↗
PubMed1995

The allosteric enhancer, PD 81,723, stabilizes human A1 adenosine receptor coupling to G proteins.

2-Amino-3-benzoylthiophenes such as PD 81,273 (PD) have been shown to increase agonist, but not antagonist, radioligand binding and to enhance functional effects of A1 adenosine receptor (A1AR) activation in tissues derived from rats and guinea-pigs. The mechanism by which PD produces this allosteric enhancement, and its effect on human A1ARs was not known. In this study, we demonstrate that PD modifies recombinant human A1AR binding and function in stably transfected CHO cells. In membranes, PD (20 microM) causes: (i) a 3-fold increase in the fraction of receptors found in a high affinity G-protein coupled conformation as assessed by the binding of [125I]N6-(3-iodo-4-aminobenzyladenosine) (125I-ABA), an A1AR agonist; (ii) a 2.44-fold increase in the potency of the agonist R-N6-phenylisopropyladenosine (R-PIA) to compete for binding with the antagonist radioligand, [3H]8-cyclopentyl-1,3-dipropylxanthine ([3H]CPX); (iii) a 1.5-fold increase in the t1/2 of 125I-ABA to dissociate from A1AR; and (i.v.) a 2.2-fold increase in the concentration of guanosine-5'-3-O-(thio)triphosphate (GTP gamma S) required to half-maximally uncouple receptor-G-protein complexes. In intact CHO cells expressing A1AR, PD increases the potency of R-PIA to decrease forskolin-stimulated cAMP accumulation by 3.3-fold. We speculate that PD binds to A1AR at a site distinct from the agonist binding site and stabilizes agonist-R-G complexes.

Allosteric Regulation↗
PubMed1995

Autosomal dominant macular dystrophy simulating North Carolina macular dystrophy.

OBJECTIVE: To characterize an autosomal dominant macular dystrophy with highly variable expression that does not fall clearly into a known disease entity. METHODS AND PATIENTS: Clinical, angiographic, and electrophysiologic data of five affected members in a family of Indian origin were evaluated. Molecular genetic analysis was undertaken to assess whether the gene responsible for the phenotype in this pedigree mapped to a region previously assigned to dominantly inherited macular dystrophies, including North Carolina macular dystrophy. RESULTS: The fundus appearance in the proband simulated stage 3 North Carolina macular dystrophy. Affected relatives had features in common with pattern dystrophy, fundus flavimaculatus with a dark choroid, and dominantly inherited drusen. Linkage to loci assigned to a number of retinal dystrophies principally affecting the posterior pole, including the North Carolina macular dystrophy locus, was excluded. CONCLUSION: The findings support the view that different genotypes are associated with similar phenotypes in autosomal dominant macular dystrophy.

Adult↗
PubMed1995

Histopathological changes induced by chronic nonlethal levels of elsan, mercury, and ammonia in the small intestine of Channa punctatus (Bloch).

Histopathological changes in the intestine of Channa punctatus induced by chronic nonlethal levels of Elsan (211 ppb), mercury (16.7 ppb), and ammonia (15.64 ppm) were studied at 7-day intervals for 90 days and the data were presented only for days (7, 28, 63, and 90) when the most conspicuous changes were noted after treatment. In the earlier phases of Elsan treatment (7 and 28 days) overall destruction of the structure of villus and other layers was prominent. Histopathology of the intestine of C. punctatus, after 63-day Elsan exposure, could be described as collapsed villi with the tips merged with each other to give a flattened appearance. Ninety-day Elsan exposure demonstrated severe damage in the longitudinal muscle layer. After 7-day mercury treatment a high degree of necrosis was indicated by submucosal area, whereas 28-day mercury treatment revealed collapsed villi due to necrotic mucosal cells and goblet cells. Mercury treatment for 63 days caused disarray of all the layers, but some improvement of villus organization was noted in fish treated with mercury for 90 days. Seven-day exposure to ammonia deteriorated the normal structure of the villus, whereas in 28-day ammonia exposure, lesions were predominant in the submucosal layers. Sixty-three-day ammonia treatment demonstrated an effect similar to that produced by Elsan, associated with a more complete destruction of all the layers. Fishes under ammonia treatment for 90 days demonstrated extensive damage to the mucosal folds. The major changes in thickness of different layers of the intestine were also evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗
PubMed1995

Gonadotropin releasing hormone stimulation of cyclic 3',5'-AMP in the pituitary cell of a teleost (Channa punctatus, Bloch) requires extracellular calcium: its relationship to gonadotropin release.

Pituitary cells were prepared by enzymatic dispersion and incubated in vitro. To observe the effect of gonadotropin releasing hormone (GnRH) and Ca2+ on the murrel pituitary cyclic 3',5'-AMP (cAMP), cells were dispersed by 0.3% collagenase plus 0.05% trypsin in Earle's minimum essential medium without Ca2+ and a considerably high yield of viable cells were obtained. Addition of a murrel, Channa punctatus, GnRH (cGnRH, 10 micrograms/incubation) to pituitary cell incubation (6 x 10(4) cells/well) containing 4 mM theophylline, a phosphodiesterase (PDE) inhibitor, stimulated cAMP accumulation in the pituitary cell 2.4-fold and its release into the medium about 2-fold as compared to control. The extent of stimulation was greatly increased on addition of Ca2+ (2 mM/incubation) with cGnRH: accumulation 5.8-fold and release 3.7-fold, respectively, in comparison to control. A time-course study with cGnRH (20 micrograms/incubation) plus Ca2+ (2 mM/incubation) on pituitary cell cAMP accumulation showed that the peak of cAMP level was reached at 15 min and remained at the same level until 60 min in the presence of theophylline; this peak was drastically reduced (5-fold) at 30 min in the absence of theophylline, indicating rapid hydrolysis of cAMP by PDE. Ca(2+)-augmented cGnRH stimulatory effect on cAMP accumulation and release could be significantly (P < 0.01) inhibited by verapamil (3 microM/incubation), a specific calcium channel blocker, suggesting requirement of extracellular Ca2+ influx in this process. Calmodulin (CaM), a Ca2+ carrier protein, addition to cGnRH and Ca2+ incubation further augmented the increase of cellular accumulation of cAMP and its release by 39.5 and 45%, respectively, in comparison to cGnRH and Ca2+ (both were statistically significant, P < 0.01). CaM effect could be blocked by calmidazolium (1 microM/incubation), a specific inhibitor of CaM, indicating specificity of the stimulatory action of CaM. Addition of radioiodinated 125I-CaM, in the presence of Ca2+ or cGnRH plus Ca2+ resulted in the binding of 125I-CaM to pituitary cells and to the pellet of the lysed cells. 125I-CaM specifically binds to pituitary cell plasma membrane preparation and saturation of 125I-CaM binding occurred at 9 ng of 125I-CaM. To investigate whether cGnRH plus Ca(2+)-stimulation of pituitary cells cAMP is linked to gonadotropin (GtH) release, similar protocols were followed. It was found that GtH release was augmented to 7-fold by cGnRH plus Ca2+, which was inhibited by verapamil and stimulated by CaM in a similar manner as observed in the case of cAMP.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Nutrient availability and the RAS/cyclic AMP pathway both induce expression of ribosomal protein genes in Saccharomyces cerevisiae but by different mechanisms.

By differential hybridization, we identified a number of genes in Saccharomyces cerevisiae that are activated by addition of cyclic AMP (cAMP) to cAMP-depleted cells. A majority, but not all, of these genes encode ribosomal proteins. While expression of these genes is also induced by addition of the appropriate nutrient to cells starved for a nitrogen source or for a sulfur source, the pathway for nutrient activation of ribosomal protein gene transcription is distinct from that of cAMP activation: (i) cAMP-mediated transcriptional activation was blocked by prior addition of an inhibitor of protein synthesis whereas nutrient-mediated activation was not, and (ii) cAMP-mediated induction of expression occurred through transcriptional activation whereas nutrient-mediated induction was predominantly a posttranscriptional response. Transcriptional activation of the ribosomal protein gene RPL16A by cAMP is mediated through a upstream activation sequence element consisting of a pair of RAP1 binding sites and sequences between them, suggesting that RAP1 participates in the cAMP activation process. Since RAP1 protein decays during starvation for cAMP, regulation of ribosomal protein genes under these conditions may directly relate to RAP1 protein availability. These results define additional critical targets of the cAMP-dependent protein kinase, suggest a mechanism to couple ribosome production to the metabolic activity of the cell, and emphasize that nutrient regulation is independent of the RAS/cAMP pathway.

Cyclic AMP↗
PubMed1995

FISH mapping of 22 novel X chromosome cosmids and the isolation of a novel microsatellite on proximal Xp.

Twenty-two X-linked cosmid clones have been localised by fluorescence in situ hybridisation (FISH). Twelve map to the long arm of the X chromosome and 10 to the short arm. Seven of the latter cosmids and an additional one were mapped by two colour FISH relative to reference markers DXS7 and DXS426 in proximal Xp. Finer localisation of one of the cosmids, namely HX43 was achieved by isolation of a microsatellite followed by genetic mapping with respect to reference markers in the region.

Base Sequence↗
PubMed1995

Ligation of endothelial alpha v beta 3 integrin increases capillary hydraulic conductivity of rat lung.

Complement-mediated pulmonary edema results from increases in lung capillary hydraulic conductivity (Lp), possibly by receptor-mediated mechanisms. We considered the Lp effects of vitronectin and the vitronectin-containing complement complex SC5b-9, which ligate the integrin alpha v beta 3. Vitronectin, SC5b-9, and SC5b-9-enriched zymosan-activated serum all rapidly increased Lp, as determined by the split-drop technique in single lung capillaries of rat lung. The Lp increases were inhibited by a monospecific (LM609) and a polyclonal (R838) antibody against the alpha v beta 3 integrin but not by an irrelevant monoclonal antibody isotype matched with LM609, by a monoclonal antibody against the alpha v beta 5 integrin, or by preimmune rabbit serum. Vitronectin monomers failed to increase Lp. The tyrosine kinase blockers genistein and methyl 2,5-dihydroxycinnamate caused significant concentration-dependent inhibitions of Lp increases due to vitronectin and zymosan-activated serum. By contrast, the protein kinase C blocker calphostin C had no major effect. We conclude that (1) multivalent ligation of the luminally located alpha v beta 3 integrin of lung capillary endothelium increases transcapillary liquid flux, and (2) the dominant signal transduction pathway for this effect occurs through tyrosine kinase activation.

Animals↗
PubMed1994

Iodized oil in the treatment of hepatocellular carcinoma.

When injected into the hepatic artery the contrast agent Lipiodol (iodized poppy seed oil) is selectively retained by hepatocellular carcinoma (HCC) for a prolonged period of time. Liver computed tomography (CT) performed after Lipiodol angiography is more sensitive than ordinary CT at imaging HCC. Arterial administration of cytotoxic drugs and radioisotopes conjugated to Lipiodol has been shown to be reasonably safe in patients with irresectable HCC. These therapies, often combined with embolization, provide effective palliation, better tumour response and improved survival compared with other available treatments. Their use as a preoperative adjunct to surgical resection of HCC is controversial.

Animals↗
PubMed1994

Drug mixture which improves survival of ischemic rabbit epigastric skin flaps.

The chief aim of this study was to maximize flap survival by counteracting the pathophysiological changes occurring during ischemia-reperfusion. Rabbit epigastric skin flaps given 21 hours of ischemia were infused intra-arterially with selected drugs at the start of reperfusion. Compared with control infused ischemic flaps, which had a 33% survival rate on day 7 post-ischemia, significant improvement was found with vasodilators nitrendipine (61%) and prostacyclin (65%) and the thrombolytic agent urokinase (65%); marginal improvement with the free radical scavenger desferrioxamine (53%); but no change with streptokinase (44%), heparin (21%), and ATP-MgCl2 (35%). A drug mixture comprising all of these agents except streptokinase and urokinase produced 87% survival, suggesting an additive effect. Biochemical assays on skin homogenates and blood implicated oxygen free radicals, neutrophil infiltration, and thromboxane in flap failure. These results imply that multiple factors are responsible for ischemic flap failure and that a mixture of drugs needs to be infused to counteract all of the detrimental changes.

Adenosine Triphosphate↗
PubMed1994

Histopathology of kidney of Channa punctatus exposed to chronic nonlethal level of Elsan, mercury, and ammonia.

Histopathological changes in the head and trunk kidneys of Channa punctatus induced by chronic nonlethal levels of Elsan (211 ppb), mercuric chloride (16.7 ppb), and aqueous ammonia (15.64 ppm) were studied on 7, 28, 63, and 90 days of exposure. The pathology of the head kidney was characterized by degeneration and dispersion of interrenal and chromaffin tissue and necrosis in the haemopoietic elements. Kidney lesions were observed throughout the entire experimental period in fish exposed to Elsan and mercuric chloride. In contrast, the lesion induced by exposure to aqueous ammonia began to heal during the first phase of treatment. Marked abnormalities in trunk kidney histology were also found. Renal lesions consisted of minimal to mild multifocal, acute tubular epithelial degeneration, karyolysis, and dilation or shrinkage of Bowman's capsule and glomerulus. Elsan treatment resulted in a highly significant decrease in the dimension of Bowman's capsule and glomerulus at all days of sampling, except on Day 28. The response of the fish trunk kidney tissue to mercuric chloride was similar to that observed with Elsan exposure in terms of the alteration in the mean dimensions of Bowman's capsule and glomerulus. The response to ammonia was significant reduction in the size of Bowman's capsule and glomerulus throughout the experimental period except at Day 28. Little dilation of Bowman's capsule and a significant dilation of glomerulus were found at Day 28 of ammonia exposure. This study demonstrated that a chronic nonlethal exposure to Elsan, mercuric chloride affect both endocrine and excretory parts of the kidney while ammonia specifically damages the excretory part of the kidney of C. punctatus.

Ammonia↗
PubMed1994

Nucleotide sequence organisation and analysis of the nuclear ribosomal DNA circle of the protozoan parasite Entamoeba histolytica.

We have sequenced the extrachromsomal ribosomal DNA (rDNA) circle of the human protozoan parasite Entamoeba histolytica HM-1:IMSS and present here the complete sequence organisation of the 24.5-kb molecule. Each circle contains two 5.9-kb rDNA transcription units organised as inverted repeats. The regions downstream (3543 bp) and upstream (9216 bp) of the rDNAs contain various families of short tandem repeats. Some of the upstream repeats share extensive sequence homology with the downstream repeats. In addition to the rDNAs themselves, the rDNA circle appears to code for only one other transcript which is 0.7 kb in size as seen in Northern blots. From DNA sequence analysis, no open reading frame could be assigned to the transcript. Extrachromosomal rDNA circles also exist in other E. histolytica strains. Restriction enzyme maps of rDNA circles were constructed from E. histolytica strains 200:NIH, HK-9 and Rahman; and Entamoeba moshkovskii strain Laredo. Striking differences were observed in the organisation of some of them, e.g. the HK-9, Rahman and Laredo circles contained only one rDNA unit and lacked the 0.7-kb transcript sequence. The short repeat sequences upstream and downstream of rDNAs were present in HK-9 and Rahman but absent in Laredo. Circles with one rDNA unit may be derived from those with two units by homologous recombination at direct repeat sequences located upstream and downstream of the two rDNAs.

Animals↗
PubMed1994

Binding of thyroid hormone to the goat testicular Leydig cell induces the generation of a proteinaceous factor which stimulates androgen release.

Leydig cells isolated from goat testis were sonicated and pure nuclear preparations obtained for 125I-3,5,3'-triiodothyronine (T3)-binding assay. Under optimum assay conditions of pH 7.2 at 37 degrees C and 90 min of incubation, binding of 125I-T3 to Leydig cell nuclei reached saturation at 1.2 nmol/l concentration. A Scatchard analysis of T3 binding exhibited a Kd of 0.535 x 10(-9) mol/l and a maximum binding capacity of 1.25 pmol/mg DNA. Competitive inhibition studies showed T3 binding to be analogue specific. The physiological relevance of T3 binding to goat Leydig cell was examined by adding increasing concentrations of T3 to the Leydig cell incubation (1 x 10(6) cells/incubation). T3 (10, 25 and 50 ng/ml or 4, 10 and 20 ng/incubation) resulted a dose dependent increase in androgen release and in all cases stimulation of androgen release was statistically significant (P < 0.01) compared with control. Stimulation of Leydig cell androgen release by T3 was significantly inhibited by actinomycin-D (P < 0.01) and cycloheximide (P < 0.01). T3 had additive stimulatory effects on LH-augmented androgen release from Leydig cells. T3 (50 ng/ml or 20 ng/incubation) effected a more than twofold increase in Leydig cell protein synthesis compared with control and both actinomycin-D and cycloheximide (50 micrograms/ml) inhibited it completely. The data indicated that the stimulatory effect of T3 on androgen release is mediated via T3-induced protein(s).(ABSTRACT TRUNCATED AT 250 WORDS)

Androgens↗
PubMed1994

Correlation of metal distribution, reduced glutathione and metallothionein levels in liver and kidney of rat.

Effect of group IIB metals on the endogenous status of metallothionein (MT) and reduced glutathione (GSH) was studied in two vital detoxifying organs namely, liver and kidney of rat. The metals were administered at non lethal levels (1/10 LD50) which were found to cause no death. Zinc showed accumulation in both liver and kidney, cadmium preferentially in the liver while mercury in the kidney. Hepatic MT content was increased by 18-fold, 15-fold and 2-fold by cadmium, zinc and mercury respectively while renal MT was increased maximally by zinc. Among the metals, mercury caused highest depletion of hepatic GSH level (51%). The renal GSH showed differential response to the metal treatment, the level increasing slightly by cadmium and depleting significantly by zinc and mercury. A positive correlation was found between group IIB metal accumulation and the manifestation of toxic response.

Animals↗