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Biomedical subjects

M Bhattacharya

Publications and source records attributed to M Bhattacharya.

At least 55 records · Page 3Linked to original sources
PubMed1998

Detection of HIV-RNA-positive monocytes in peripheral blood of HIV-positive patients by simultaneous flow cytometric analysis of intracellular HIV RNA and cellular immunophenotype.

Determinations of plasma HIV viral RNA copy numbers help to define the kinetics of HIV-1 infection in vivo and to monitor antiretroviral therapy. However, questions remain regarding the identity of various infected cell types contributing to this free virus pool and to the in vivo lifecycle of HIV during disease progression. Characterization of a novel fluorescence in situ hybridization (FISH) assay employing a pool of labeled oligonucleotide probes directed against HIV RNA was done followed by coupling of the FISH assay with simultaneous surface immunophenotyping to address these questions. In vitro characterizations of this assay using tumor necrosis factor-alpha stimulated and unstimulated ACH-2 cells demonstrated the ability to detect < 5% HIV RNA positive cells with a sensitivity of < 30 RNA copies per cell. Peripheral blood mononuclear cells from 39 HIV-seropositive patients on no, single, combination, or triple drug therapy and 8 HIV-seronegative patients were examined. The majority of HIV-positive patients (24/39) harbored monocytes positive for HIV RNA and a significantly higher fraction of patients with high plasma viral load carried positive monocytes (13/16) than did patients in the low plasma viral load group (11/23). These results demonstrate the effectiveness of a novel FISH assay for identifying and monitoring HIV-infected cell populations in the peripheral blood of HIV-positive patients. In addition, monocytes are a major source of cellular HIV virus in the peripheral blood of HIV patients, even with progression of disease.

Cell Line↗
PubMed1998

Immunosuppressant activity in human beta-casein fragment analogs.

Human beta-casein fragment (54-59) having the amino acid sequence Val-Glu-Pro-Ile-Pro-Tyr, has been shown potent immunostimulant activity. Several analogs of this hexapeptide have been synthesized with modification in the N-terminal region and tested for their immunomodulatory activity. Interestingly, two hexapeptides have shown significant immunosuppressant activity.

Amino Acid Sequence↗
PubMed1998

Expression of cyclooxygenases in ductus arteriosus of fetal and newborn pigs.

OBJECTIVE: We studied the ontogeny of the expression of cyclooxygenase-1 and cyclooxygenase-2 in the ductus arteriosus and evaluated their functional significance. STUDY DESIGN: The expression of cyclooxygenase-1 and cyclooxygenase-2 was studied in ductus arteriosus of fetal (at approximately 75% gestation) and term newborn pig. Effects of selective inhibitors of cyclooxygenase-1 and cyclooxygenase-2 on ductal patency were evaluated by Doppler ultrasonography. RESULTS: Ductus arteriosus of the fetus expressed virtually only cyclooxygenase-1 immunoreactive protein and activity. In contrast, the ductus of the term newborn pig (<45 minutes old) contained proteins of both cyclooxygenase-1 and cyclooxygenase-2, but the latter contributed to >90% of prostaglandin E2 formation. The selective cyclooxygenase-2 inhibitor DuP697 reduced prostaglandin E2 levels in the ductus arteriosus, albeit not in plasma, but did not affect ductal patency in the newborn pig (<1(1/2) hours old); in contrast, the cyclooxygenase-1 inhibitor valeryl salicylate, like indomethacin, markedly reduced levels of prostaglandin E2 in the plasma and ductus arteriosus and caused significant constriction of the ductus arteriosus. CONCLUSION: The ductus arteriosus of the term newborn pig, in contrast to that of the fetus, expresses cyclooxygenase-2, but circulating prostaglandins, arising mostly from cyclooxygenase-1, seem to exert the major control on ductal patency in vivo. Our data suggest that cyclooxygenase-2 inhibitors might be better alternatives for the fetus than nonselective cyclooxygenase blockers if indicated for maternal conditions such as inflammation or for tocolysis.

Animals↗
PubMed1998

Cross-contamination of specimens with Mycobacterium tuberculosis: clinical significance, causes, and prevention.

At the Veterans Affairs Lakeside Medical Center, two episodes of specimen cross-contamination with Mycobacterium tuberculosis were detected during a 54-month period by molecular strain typing using DNA restriction fragment length polymorphism for 3 patients without clinical or radiologic signs of tuberculosis (TB). A cross-contaminated specimen was the only culture-positive specimen for each of the 3 patients. Laboratory features of cross-contamination included acid-fast smear negativity, growth only in broth or solid medium, and growth in solid medium with 5 or fewer colonies. Retrospective analysis demonstrated identical features for occasional culture-positive specimens from 54 patients with TB during the same period. However, productive cough, pleural pain, weight loss, night sweats, chest radiograph results suggestive of TB, positive tuberculin skin testing, and/or multiple culture-positive specimens were invariably present in patients with TB with such specimens. Most patients with TB (50/54; 93%) had multiple specimens positive in culture for M. tuberculosis, and the few patients with TB with single culture-positive specimens were symptomatic. These results indicate that correlation with clinical manifestations is necessary to determine the significance of isolated, acid-fast smear negative, and/or low-yield culture-positive specimens. Although the prevalence of specimen cross-contamination is low (0.1%), possible sources (especially the use of single-reagent delivery systems for multiple specimens) should be eliminated by mycobacteriology laboratories.

Adult↗
PubMed1998

Increases in retinovascular prostaglandin receptor functions by cyclooxygenase-1 and -2 inhibition.

PURPOSE: To determine the relative contribution of cyclooxygenase (COX)-1 and COX-2 in regulating prostaglandin (PG) E2 and PGF2alpha receptors (EP and FP, respectively) densities and their functions in retinal vasculature of neonatal pigs. METHODS: Newborn pigs were treated intravenously every 8 hours for 48 hours with saline, 40 mg/kg nonselective COX inhibitor ibuprofen, 80 mg/kg COX-1 inhibitor valeryl salicylate, or 5 mg/kg DuP697 and 5 mg/kg NS398, COX-2 inhibitors. Retinal microvessel EP and FP receptor densities were measured by radioligand binding and receptor-coupled effects by determining second-messenger inositol 1,4,5-trisphosphate (IP3) and vasomotor responses. Retinal blood flow (RBF) response to incremental increases in blood pressure (BP) was measured by a microsphere technique. RESULTS: Valeryl salicylate, DuP697, and NS398 reduced retinal PGE2 and PGF2alpha concentrations in the newborn by approximately half, whereas ibuprofen caused further reduction to levels observed in adults. Retinal vessel EP1, EP3, and FP receptor densities increased approximately threefold after treatments with COX-1 or COX-2 inhibitors, and five- to sixfold after ibuprofen treatment. EP and FP receptor upregulation was associated with corresponding increases in IP3 production and retinal vasoconstriction in response to PGF2alpha, fenprostalene (an FP agonist), PGE2, 17-phenyl trinor PGE2 (an EP1 agonist), and M&B28,767 (an EP3 agonist) and with enhanced RBF autoregulation of high BP (> or =125 mm Hg). Conversely, EP2 receptor density and coupled functions were minimally affected by COX inhibition. CONCLUSIONS: Data suggest that increased COX-1- and COX-2-catalyzed prostaglandin synthesis contribute equivalently to the downregulation of retinovascular EP1, EP3, and FP receptors and their vasoconstrictor functions in newborn pigs; the EP2 receptor was not significantly influenced by ontogenic alterations in prostaglandin levels.

Animals↗
PubMed1997

Prevention of postasphyxia electroretinal dysfunction with a pyridoxal hydrazone.

The newborn retina is particularly sensitive and frequently subjected to peroxidative stresses that result in visual sequelae. We compared two iron chelators, deferoxamine and a newer compound, pyridoxal isonicotinoyl hydrazone (PIH), in protecting the retina of newborn pigs (1-3 d old) from asphyxia-reoxygenation insults. Animals were treated IV with either saline, deferoxamine 15.2 mumol/kg (10 mg/kg) or PIH 34.8 mumol/kg (10 mg/kg); n = 10 in each treatment group. Scotopic and photopic electroretinograms (ERG) were recorded before and 40 min after drug treatment as well as 45 min following a 5-min period of asphyxia by interrupting ventilation. In separate animals the indices of peroxidation, malondialdehyde (MDA: TBARS) and hydroperoxides, were measured in retina at the same times. In saline-treated animals, there was a marked increase in MDA and hydroperoxide concentrations in the retina following the asphyxia-reoxygenation period. This was associated with a decrease in the a- (photoreceptor generated) and b-wave (generated by Müller and bipolar cells) amplitudes measured under photopic (cone-mediated response) and scotopic (rod-mediated response) conditions, and an increase in their implicit times. PIH and deferoxamine prevented the postasphyxial increase in MDA and hydroperoxides. However, only PIH prevented the postasphyxial changes in a- and b-wave amplitudes and implicit times, whereas deferoxamine markedly altered the preasphyxial ERG and provided only partial postasphyxial protection simply to the retinal outer segment. Our findings indicate that the iron chelator PIH effectively inhibits peroxidation and retinal electrophysiological alterations secondary to asphyxia-reoxygenation-induced oxidative stresses to newborn animals, whereas deferoxamine adversely affects retinal function; hence, PIH may be a preferred alternative to deferoxamine.

Animals↗
PubMed1997

Key role for cyclooxygenase-2 in PGE2 and PGF2alpha receptor regulation and cerebral blood flow of the newborn.

Ibuprofen, a cyclooxygenase (COX) inhibitor nonselective for either COX-1 or COX-2 isoform, upregulates cerebrovascular prostaglandin E2 (PGE2) and PGF2alpha receptors in newborn pigs. COX-2 was shown to be the predominant form of COX and the main catalyst of prostaglandin synthesis in the newborn brain. We proceeded to establish direct evidence that COX-2-generated prostaglandins govern PGE2 and PGF2alpha receptor density and function in the cerebral vasculature of the newborn. Hence, we determined PGE2 and PGF2alpha receptor density and functions in brain vasculature by using newborn pigs treated with saline, ibuprofen, COX-1 inhibitor (valerylsalicylate), or COX-2 inhibitors (DUP-697 and NS-398). Newborn brain PGE2 and PGF2alpha concentrations were significantly reduced by ibuprofen, DUP-697, and NS-398 but not by valerylsalicylate. In newborn pigs treated with DUP-697, NS-398, and ibuprofen, PGE2 and PGF2alpha receptor densities in brain microvessels were increased to adult levels; there was also a significant increase in inositol 1,4,5-trisphosphate (IP3) production and cerebral vasoconstrictor effects of 17-phenyl trinor PGE2 (EP1 receptor agonist), M&B-28767 (EP3 receptor agonist), PGF2alpha, and fenprostalene (PGF2alpha analog). Treatment with ibuprofen or DUP-697 also increased the upper blood pressure limit of cerebral cortex and periventricular blood flow autoregulation from 85 to > or = 125 mmHg (uppermost blood pressure studied). However, valerylsalicylate treatment did not affect cerebrovascular PGE2 and PGF2alpha receptors, IP3 production, or vasoconstrictor effects in newborn animals. These in vivo and in vitro observations indicate that COX-2 is mainly responsible for the regulation of PGE2 and PGF2alpha receptors and their functions in the newborn cerebral vasculature.

Animals↗
PubMed1997

Validity of using slide positivity rate (SPR) in identification of high risk malarious segments in rural areas.

The Expert Committee Report-1995, recommended various parameters for identifying worst malaria affected segments in rural areas, which are mainly based on slide positivity rate (SPR). A analysis of the epidemiological data of Uttar Pradesh was carried out in different settings to study the sensitivity of SPR in identifying high risk areas. The validity of using SPR in the entire range in all settings is examined and appropriate corrective measures suggested.

Blood Specimen Collection↗
PubMed1996

(-)-Epigallocatechin gallate, a polyphenolic tea antioxidant, inhibits peroxynitrite-mediated formation of 8-oxodeoxyguanosine and 3-nitrotyrosine.

Reaction with peroxynitrite at pH 7.4 and 37 degrees C was found to increase the 8-oxodeoxyguanosine levels in calf thymus DNA 35- 38-fold. This oxidation of deoxyguanosine, as well as the peroxynitrite-mediated nitration of tyrosine to 3-nitrotyrosine, was significantly inhibited by ascorbic acid, glutathione and (-)-epigallocatechin gallate, a polyphenolic antioxidant present in tea. For 50% inhibition of the oxidation of deoxyguanosine to 8-oxodeoxyguanosine, 1.1, 7.6 or 0.25 mM ascorbate, glutathione or (-)-epigallocatechin gallate, respectively, was required. For 50% inhibition of tyrosine nitration, the respective concentrations were 1.4, 4.6 or 0.11 mM. Thus, (-)-epigallocatechin gallate is a significantly better inhibitor of both reactions than either ascorbate or glutathione. Reaction of (-)-epigallocatechin gallate with peroxynitrite alone resulted in the formation of a number of products. Ultraviolet spectra of two of these suggest that the tea polyphenol and/or its oxidation products are nitrated by peroxynitrite.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1996

Hyperbaric oxygen therapy in parenchymal liver disease.

Use and efficacy of hyperbaric oxygen therapy (HOT) in liver disease has not been established. A prospective control study of HOT in liver diseases was undertaken. Sixty cases were selected for this study (30 with HOT and 30 with conventional therapy only). Almost equal number of Hepatitis B virus surface antigen (HbsAg) positive cases were included in both groups. All patients were male. Only cases with serum bilirubin over 10 mg/dl were included in this study. It was found that study cases recovered faster, gained appetite and had an earlier sense of well-being. There was faster disappearance of pruritus, earlier achievement of normal liver function, HbsAg negativity and overall shorter duration of hospital stay and convalescence. Short term adverse effects were unremarkable.

Adolescent↗
PubMed1996

A community-based study on the effect of hexachlorocyclo-hexane (HCH) exposure in spraymen and general population.

The study was carried out in Allahabad district, (Uttar Pradesh) with 260 spraymen as test subjects and 50 persons as controls from a sprayed and unsprayed village respectively. Majority of the spraymen (44%) had worked for 3-4 years (seasons) and 31% had worked in the programme for 5-10 years. The spraymen were relatively healthy with no complaints in 77% whereas the figures were 76% for the Community living in the sprayed village, and 50% for the Community in the unsprayed village. A comparison of the biochemical parameters revealed lowered Cholesterol more than 150 mg % in 38% of the spraymen and 58% had altered A:G ratio. Other biochemical estimations were not significantly different from the control population. The mean residue of Alpha, Beta and Gamma Isomers and the total Alpha, Beta and Gamma isomers were 0.0317, 0.2254, 0.0288 and 0.2859 mg/1 respectively; the corresponding mean values in the control population were 0.0211, 0.1112, 0.0197 and 0.1520 mg/1 respectively. The values in spraymen were twice those of the general population. A significant association (p < .05) was observed between their length of exposure and the levels of Cholesterol and HCH isomers in blood of spraymen. No significant morbidity was evident in spraymen due to HCH exposure.

Case-Control Studies↗
PubMed1995

Single-run separation and detection of multiple metabolic intermediates by anion-exchange high-performance liquid chromatography and application to cell pool extracts prepared from Escherichia coli.

A method is described for analysis of the intracellular concentrations of metabolic intermediates of the Embden-Meyerhof-Parnas pathway, the Entner-Doudoroff pathway, the pentose phosphate pathway, and the tricarboxylic acid cycle in cell pool extracts of Escherichia coli. A single anion-exchange HPLC run of 40 min allowed resolution of 27 anionic metabolite standards. Detection limits of 0.1 nmol per injection were achieved by use of a conductivity detector equipped with an anion self-regenerating suppressor and a uv detector. A boiling water extraction procedure was used to prepare cell pool extracts. Cochromatography of cell pool extracts and metabolite standards was used to confirm the identities of metabolites in the cell pool. As many as 16 metabolites could be detected and quantified in the cell pool extracts by using the described HPLC method. An analysis of metabolite concentrations in E. coli showed the dynamics of glucose metabolism during a 2-min transition from starvation to steady-state metabolism following addition of glucose. The ease and power of this method suggests general utility for in vivo metabolite analysis in a variety of experimental systems.

Carbohydrates↗
PubMed1995

The value of cervical cytology in HIV-infected women.

The purpose of this study was to assess the ability of cytology to predict the results of colposcopically directed cervical biopsies in HIV-infected women. We performed a case-control study of 52 HIV(+), 31% of whom had AIDS, and 57 HIV(-) women referred to two tertiary care centers for colposcopy from July 1991 to November 1993. All 57 HIV(-) controls and 27 HIV (+) cases underwent colposcopy for evaluation of an abnormal Pap smear. The remaining HIV(+) cases (n = 25) had colposcopy as part of their routine assessment. In women with abnormal Pap smears, colposcopic biopsy agreed with the Pap smear results in 83% of 24 HIV(+) women and 65% of 37 controls (chi 2; P = 0.34). For patients with low-grade SIL on Pap smear, 14% of HIV(+) and 11% of HIV(-) women had moderate or severe dysplasia on biopsy (P = 0.52). The positive predictive value of an abnormal Pap smear was 96% in HIV(+) women vs 78% in noninfected patients (P = 0.05). In the overall series of 52 HIV(+) women, the Pap smear did not match the biopsy in 44% of patients and was less severe than the cervical biopsy results in 91% of these mismatches. The Pap smear had a sensitivity of 57%, a specificity of 92%, a positive predictive value of 96%, and a negative predictive value of 39%, when compared to biopsy results in HIV-seropositive patients. Pap smears missed 43% of biopsy-proven intraepithelial lesions in this series of HIV (+) women. However, when abnormal, the Pap smear was no worse in predicting the presence and degree of an intraepithelial lesion in HIV(+) women than in noninfected women. These characteristics may justify immediate treatment of HIV(+) women at the time of colposcopy after an abnormal Pap smear given its high positive predictive value.

Adult↗
PubMed1993

Evidence for virus-encoded glycosylation specificity.

Four spontaneously derived serologically distinct classes of mutants of the Paramecium bursaria chlorella virus (PBCV-1) were isolated using polyclonal antiserum prepared against either intact PBCV-1 or PBCV-1-derived serotypes. The oligosaccharide(s) of the viral major capsid protein and two minor glycoproteins determined virus serological specificity. Normally, viral glycoproteins arise from host-specific glycosylation of viral proteins; the glycan portion can be altered only by growing the virus on another host or by mutations in glycosylation sites of the viral protein. Neither mechanism explains the changes in the glycan(s) of the PBCV-1 major capsid protein because all of the viruses were grown in the same host alga and the predicted amino acid sequence of the major capsid protein was identical in the PBCV-1 serotypes. PBCV-1 antiserum resistance is best explained by viral mutations that block specific steps in glycosylation, possibly by inactivating glycosyltransferases.

Agglutination↗
PubMed1993

Nonenzymatic Glycosylation of Lepidopteran-Active Bacillus thuringiensis Protein Crystals.

We used high-pH anion-exchange chromatography with pulsed amperometric detection to quantify the monosaccharides covalently attached to Bacillus thuringiensis HD-1 (Dipel) crystals. The crystals contained 0.54% sugars, including, in decreasing order of prevalence, glucose, fucose, arabinose/rhamnose, galactose, galactosamine, glucosamine, xylose, and mannose. Three lines of evidence indicated that these sugars arose from nonenzymatic glycosylation: (i) the sugars could not be removed by N- or O-glycanases; (ii) the sugars attached were influenced both by the medium in which the bacteria had been grown and by the time at which the crystals were harvested; and (iii) the chemical identity and stoichiometry of the sugars detected did not fit any known glycoprotein models. Thus, the sugars detected were the product of fermentation conditions rather than bacterial genetics. The implications of these findings are discussed in terms of crystal chemistry, fermentation technology, and the efficacy of B. thuringiensis as a microbial insecticide.

Journal Article↗