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Biomedical subjects

P Chakraborty

Publications and source records attributed to P Chakraborty.

At least 19 recordsLinked to original sources

Inhibition of volume-regulated anion channels in cultured endothelial cells by the anti-oestrogens clomiphene and nafoxidine.

1. We have used the whole-cell patch clamp technique to study the effect of the partial anti-oestrogens clomiphene and nafoxidine, the pure anti-oestrogens ICI 182,780 and RU 58,668 and the oestrogen ss-estradiol, on the volume-regulated anion channel (VRAC) in cultured pulmonary artery endothelial (CPAE) cells. 2. In contrast to the pure anti-oestrogens and ss-estradiol, clomiphene and nafoxidine potently inhibited the volume-sensitive chloride current, I(Cl,swell), activated by challenging CPAE cells with a 25% hypotonic solution. For clomiphene, the estimated IC(50) and Hill coefficient were 1.03+/-0.14 microM and 1.40+/-0.21 respectively. In the case of nafoxidine, these values were 1.61+/-0.29 microM and 1.24+/-0.19. 3. The inhibition induced by the pure enantiomers of clomiphene, zuclomiphene and enclomiphene, was not different from that of the racemic mixture, indicating that the interaction between clomiphene and VRAC is not stereoselective. 4. Clomiphene and nafoxidine inhibited proliferation of CPAE cells. Half-maximal inhibition was found at 1.98+/-0.17 and 1.66+/-0.21 microM respectively, concentrations similar to those for half-maximal block of VRAC. 5. In conclusion, the nonsteroidal partial anti-oestrogens nafoxidine and clomiphene are potent inhibitors of volume-regulated anion channels. The inhibition by clomiphene is not stereoselective and occurs at concentrations close to therapeutically relevant concentrations. Finally, both drugs inhibit the proliferation of endothelial cells.

Cell Division↗

Biogeochemical cycling bacteria as indices of pond fertilization: importance of CNP ratios of input fertilizers.

AIMS: The influence of carbon-nitrogen and nitrogen-phosphorus ratios of input fertilizers, and that of pond water, on the growth of heterotrophic and phosphate-solubilizing bacteria of water and sediment, was examined in relation to fertilizer mineralization indices using different modes of fertilization through inorganic and organic sources. METHODS AND RESULTS: The first experiment used carbon-nitrogen-phosphorus ratios varying from 12 : 2 : 1 to 151 : 6 : 1, applied at the rate of 0.043 g l(-1) week(-1), whereas in the second ratios varied from 25.6 : 6.2 : 1 to 150 : 12 : 1 applied once at the rate of 3.33 g l(-1). Different fertilizers (cattle dung, poultry droppings, urea, single superphosphate and starch) were mixed in different proportions to achieve the desired carbon-nitrogen-phosphorus ratio. The heterotrophic and phosphate-solubilizing populations were more responsive to an early manuring phase than later, implying that pond fertilization was microbiologically more dynamic in the earlier phase. The carbon-nitrogen and nitrogen-phosphorus ratios of 11.8 (88.6 : 7.5) and 7.5 (7.5 : 1), respectively, of input fertilizers favoured growth of both heterotrophic and phosphate-solubilizing bacterial populations much better than the other ratios tested. Likewise, water carbon-nitrogen and nitrogen-phosphorus ratios of 11.9 and 3.34 induced bacterial growth. The carbon-nitrogen ratios of 12.63 (101 : 8) (input fertilizer)-4.54 (water), and nitrogen-phosphorus ratios of 8 (8 : 1) (input fertilizer)-2.93 (water), gave gross primary productivity values higher than the remaining ratios, exhibiting overall curvilinear relationships. The values of gross primary productivity were the direct function of values of fertilizer mineralization indices for carbon, nitrogen and phosphorus. CONCLUSION: It is concluded that the mixed fertilizer (carbon-nitrogen-phosphorus-88.6 : 7.5 : 1) comprising cattle dung (95%), poultry droppings (2.5%), urea (2%) and single superphosphate (0.5%), applied at the rate of 23,000 kg ha(-1) year(-1), was a suitable cost-effective fertilization option for aquaculture practices. SIGNIFICANCE AND IMPACT OF THE STUDY: As chemical fertilizers are expensive and cause some adverse effects on the soil structure, composition, microflora and other characteristics of the pond, mixed combinations of inorganic and organics with narrow range of carbon-nitrogen-phosphorus ratio can be suitable and cost-effective fertilization tools in aquaculture practices, which is to be linked with the microbial activities of the pond.

Animals↗

G-protein-mediated signaling and its control in macrophages and mammalian cells.

G-protein coupled receptor (GPCR) in various cell types exert its effects through heterotrimetic GTP-binding proteins (G-proteins). The interaction of specific ligand or agonists with CPCR transuces signal and enhances gene expression, mitogen activated protein kinase (MAP kinase) activation, and thus regulates cell proliferation, differentation, and motility. Abnormal signaling or prolonged activation of G-protein signaling pathways blocks normal functioning of various cells and tissues of our body. New insights into the mechanisms governing the specificity and temporal regulation of G-protein signaling pathways have been provided by the recent discovery of GTPase-activating proteins (GAPs) and RGS proteins (regulators of G-protein signaling). Different molecular biological approaches are now being employed to study the G-protein-mediated signaling and its control in various mammalian cells. Recent developments on the activation of phagocytic cells, especially macrophages, via ligation or cross-linking of GPCR and their postreceptor ligation effect against several intramacrophage pathogens are also discussed.

Animals↗

Training of providers in embryo transfer: what is the minimum number of transfers required for proficiency?

BACKGROUND: Embryo transfer represents one of the most critical procedures in the practice of assisted reproduction. The objective of this study was to identify retrospectively the minimum number of embryo transfers required to train providers properly in this skill. METHODS AND RESULTS: The study group consisted of 204 patients who received embryo transfers between January 1996 and March 2000 in a university-based programme of assisted reproduction. The main outcome measure was clinical pregnancies per embryo transfer. Five Fellow trainees performed a total of 204 embryo transfers for an overall pregnancy rate of 45.5% per embryo transfer (93/204). In comparison, the programme pregnancy rate per transfer for experienced providers was 47.3% (560/1179). A chronological graph of each individual trainee's experience for the first 50 embryo transfers performed suggested a lower initial pregnancy rate for three of the five trainees. To determine whether a learning curve might exist, results of the first 25 transfers were compared as a subgroup with the second 25 transfers. Pregnancy rates were lower for the 1-25 transfer subgroup than in the 26-50 subgroup for three of the five Fellow trainees, although the difference was not statistically significant. CONCLUSION: Clinical pregnancy rates of Fellows-in-training were indistinguishable statistically from those of experienced staff by 50 transfers.

Adult↗

Modulation of macrophage mannose receptor affects the uptake of virulent and avirulent Leishmania donovani promastigotes.

The effect of oxidants and the anti-inflammatory steroid dexamethasone on the attachment and internalization of virulent and avirulent Leishmania donovani promastigotes by the macrophage mannosyl fucosyl receptor was examined. Oxidants and dexamethasone are known to down- and upregulate the expression of the mannose receptor. Macrophages, when treated with 500 microM H2O2 at 37 C for 30 min, stimulate about 45% inhibition in uptake of an avirulent strain (UR6), and 30 and 25% inhibition for virulent strains AG-83 and GE-I, respectively. Treatment of macrophages with dexamethasone for 20 hr resulted in a stimulation in uptake of the parasite. When UR6 was used, a 3-fold increase in uptake was observed compared with the controls. Parasite uptake was also inhibited by the H2O2-generating system, glucose/glucose oxidase; inhibition was blocked by catalase. Treatment of macrophages either with H2O2 or dexamethasone did not affect the binding of the advanced glycosylation end product-bovine serum albumin (AGE-BSA), the ligand for AGE receptor of macrophages. Similarly, indirect evidence also shows that both types 1 and 3 complement receptors (CR1, CR3) are not affected by these treatments, indicating that, besides the mannosyl fucosyl receptor, other receptors are minimally altered in the identified condition. These results suggest that the up- and downregulation of the mannose receptor of macrophages may play a role in affecting L. donovani infection.

Animals↗

Differences in concentrations of allergenic pollens and spores at different heights on an agricultural farm in West Bengal, India.

The aim of the study was to assess the vertical profile of the major airborne pollen and spore concentration in the lower heights (up to six meters) and to check their allergenic potential causing respiratory allergy in agricultural workers. The study was conducted using rotorod samplers mounted at different heights at weekly intervals for two consecutive years (November 1997-October 1999). The major pollen grains and fungal spores (from mass culture) were collected in bulk and studied by skin-prick tests to detect allergenicity. Of the recorded pollen, 10 major and perennial types (e.g., Poaceae, Cheno-Amaranthaceae, Cyperaceae, Areca, etc.) were considered for comparative analyses. The tree pollen count showed more or less good correlation with increasing heights, whereas herb/shrub members are dominant at lower heights during all the three seasons (winter, summer and rains). The 10 major and perennial fungal spore types included Aspergilli group, Cladosporium, Nigrospora, etc. The smaller spores were dominant at greater heights and larger spores and conidia were more prevalent at lower levels. The total spore count was higher just after the rainy season during winter. In terms of allergenicity, Saccharum officinarum (sugar cane) of Poaceae, showed highest reactivity (70.58%) in skin test carried out in 189 adult agricultural field workers with respiratory disorders living inside the study area. Among fungal spores, Aspergillus japonicus was the strongest allergen, evoking 74.07% positive reactions. Drechslera oryzae, the pathogen causing brown spot of rice was also found to be a potent allergen.

Adolescent↗

Binding of globular proteins to DNA from surface tension measurement.

Extent of binding (gammap) of globular proteins to calf-thymus DNA have been measured in mole per mole of nucleotide as function of equilibrium protein concentration. We have exploited measurement of the surface tension of the protein solution in the presence and absence of DNA to calculate the binding ration (gammap). Interaction of bovine serum albumin with DNA has been studied at different pH. Interaction of bovine serum albumin with DNA has been studied at different pH, ionic strength and in presence of Ca2+. Interaction of BSA with denatured DNA has also been investigated. Binding isotherms for other globular proteins like beta-lactoglobulin, alpha-lactalbumin and lysozyme have been compared under identical physicochemical condition. It has been noted with considerable interest that globular form of protein is important to some extent in protein-DNA interaction. An attempt has been made to explain the significance of difference in binding ratios of these two biopolymers in aqueous medium for different systems in the light of electrostatic and hydrophobic effects. Values of maximum binding ration (gammap(m)) at saturated level for different systems have been also presented. The Gibb's free energy decrease (-deltaG0) of the binding of proteins to DNA has been compared more precisely for the saturation of binding sites in the DNA with the change of activity of protein in solution from zero to unity in the rational mole fraction scale.

Animals↗

Evaluation of sympathetic nerve terminals with [(11)C]epinephrine and [(11)C]hydroxyephedrine and positron emission tomography.

BACKGROUND: The goal of the present study was to directly compare the new radiopharmaceutical agent [(11)C]epinephrine (EPI) with [(11)C]hydroxyephedrine (HED) through the use of PET. METHODS AND RESULTS: Seven healthy volunteers and 10 patients were investigated after heart transplantation. PET images of both tracers were of excellent quality in the volunteers. Values for radiolabeled metabolites (measured in percent of blood activity) at 5, 20, and 60 minutes after injection were approximately 35%, approximately 82%, and approximately 86% for EPI and approximately 13%, approximately 47%, and approximately 78% for HED, respectively. At 35 minutes, metabolite-corrected mean myocardial retention fraction of EPI (0. 235+/-0.022 min(-1)) was significantly greater (P<0.01) than that of HED (0.142+/-0.012 min(-1)). Corrected tracer retention fractions of both EPI and HED were significantly reduced in transplant recipients (0.055+/-0.004 min(-1), P<0.0001; and 0.050+/-0.006 min(-1), P<0. 0001, respectively) compared with volunteers. Normalization of retention fractions of patients with transplantation within 1 year to volunteers resulted in a value (ratio expressed in percent) of 20. 6+/-1.8% for EPI, significantly (P<0.03) smaller than 27.8+/-0.8% for HED. In patients with transplantation later than 1 year, the values were 26.0+/-2.9% for EPI compared with 44.2+/-5.6% for HED (P<0.014). CONCLUSIONS: Both tracers showed high selectivity for neuronal uptake in the heart, with a significant reduction in tracer retention in transplant recipients compared with volunteers. Compared with HED, EPI showed greater retention in volunteers and a lower retention ratio in transplant recipients, suggesting that EPI may be the superior tracer with higher sensitivity to neuronal abnormalities. Because EPI reflects neuronal uptake, metabolism, and storage, it may be more suitable for the study of neuronal integrity than HED, which primarily traces uptake-1 capacity.

Adult↗

Ligation of Fc receptor of macrophages stimulates protein kinase C and anti-leishmanial activity.

Fc receptors are known to express on the surface of mature monocytes, macrophages and lymphocytes. In this study a ligand e.g. liposomal IgG (human IgG coupled to PE-liposome via carbodimide reaction) was developed to ligate the Fc receptor of macrophages. When liposomal IgG was incubated with macrophages at 37 degrees C for 5 min, it induced the macrophage activation which suppress the parasite burden approximately to an extent of 60%, 50% and 45%, when macrophages were infected with UR6, AG83 and GE1 strains of L-donovani respectively. Superior efficacy of liposomal IgG were achieved compared to the treatment with free IgG and free liposomes. The activity of protein kinase C (PKC) has been found to be higher in the Fc receptor targeted macrophage membrane fraction, suggesting its translocation from the cytosol. Staurosporine, a potent inhibitor of the enzyme protein kinase C (PKC) has been found to protect the parasite inside the macrophage indicating the role of PKC in the signaling process. The liposomal IgG treatment has been found to induce the generation of significant amount of superoxide and hydrogen peroxide which helped to suppress the parasite burden. Further when liposomal IgG were incubated with IFN-gamma primed, LPS activated macrophages, a significant amount of NO release was also noticed, indicating its role in parasite killing. The above results suggest that Fc receptor mediated activation by liposomal IgG may be used as an alternative approach to kill parasites intracellularly.

Animals↗

fMLP receptor stimulated activation of macrophage: its effect on killing of intracellular Leishmania donovani.

The fMLP receptor of peritoneal macrophages stimulated by fMLP grafted liposomes as ligand, was analysed and compared with respective controls for its ability to promote killing of intracellular Leishmania parasites. fMLP grafted liposomes show greater efficacy in killing intracellular L. donovani (MHOM/IN/1983/AG83) parasites in a time dependent manner than free fMLP. fMLP grafted liposomes also release more active oxygen intermediates and reactive nitrogen intermediates (O2-, H2O2, NO) than free fMLP. The key enzymes PKC and PTK for the respiratory burst and nitric oxide generation were found to be important in this fMLP receptor mediated signaling process as the enzyme specific inhibitors viz. staurosporine, genistein and AG126 suppressed the leishmanicidal effect of fMLP grafted liposomes. The above findings suggest that the fMLP receptor of macrophages activates PKC and PTK mediated signalling that is responsible for the intracellular parasite killing.

Animals↗

Macrophage protein kinase C: its role in modulating membrane microviscosity and superoxide in leishmanial infection.

Pretreatment of macrophages with, an agonist of PKC, showed diverse effects on degradation and survival of two virulent strains of Leishmania donovani promastigotes. Treatment of macrophages with PMA for 45 min at 37 degrees C generated significant amounts of superoxide anions and reduced the parasite burden of macrophages by up to 48 and 43% when AG83 and GE-1 strains were used for infection. Staurosporine, an inhibitor of PKC, inhibited PMA-dependent killing of the parasites, while tyrphostin AG 126, an inhibitor of protein tyrosine kinase, showed very little effect. Depletion of PKC by prolonged incubation with PMA drastically reduced the superoxide anion generation and increased the uptake and multiplication of the parasites. Finally, to understand the mechanism of higher uptake of the parasites by PKC-depleted macrophages, membrane microviscosity was measured by fluorescence depolarization. Membrane microviscosity was found to be approximately 40% lower in PKC-depleted macrophages than in normal macrophages, indicating the role of membrane fluidity in the infection process. Together, these data suggest PKC activation, superoxide generation, and membrane fluidity are essential factors in the efficient regulation of leishmanial infection.

Animals↗

Clinicoimmunologic studies on Phoenix sylvestris Roxb. pollen: an aeroallergen from Calcutta, India.

BACKGROUND: This study highlights the allergenicity and allergenic components of the pollen of Phoenix sylvestris Roxb. (PS), or date sugar palm, which is predominantly airborne in the air of Greater Calcutta. METHODS: A 2-year aerobiologic survey was performed by Burkard sampler. PS pollen extract was used in skin tests of allergic patients, fractionated by (NH4)2SO4 and the Sephacryl S-200 column. The allergenicity of each fraction was checked by skin test and IgE ELISA inhibition. The principal allergenic fraction, Fr.lla, was separated in 11% SDS-PAGE, and its allergenicity was confirmed by IgE ELISA inhibition and immunoblotting. RESULTS: PS pollen grains were found to be prevalent in the air of the suburban zone of Calcutta from January to March with a peak in February. The pollen extract showed high (44.07%) positive skin reaction on 540 respiratory allergic patients. Among the (NH4)2SO4 cut fractions, Fr.II was the most active one, and it was resolved into four subfractions in the Sephacryl S-200 column. Fr.lla was the principal allergenic fraction, showing the presence of two components of 33 and 66 kDa in SDS-PAGE. In IgE immunoblotting, both of the components were found to be allergenic. CONCLUSIONS: The PS pollen grain is an important aeroallergen from Calcutta, India. The 33- and 66-kDa components are the major allergens present in the relevant pollen extract.

Allergens↗

Antigenic relationship between four airborne palm pollen grains from Calcutta, India.

The pollen grains of Areca catechu, Borassus flabellifer, Cocos nucifera and Phoenix sylvestris, all belonging to the family Aracaceae (Palmae), are airborne and found to be potent in causing human respiratory allergy. The present study was undertaken to discover the antigenic relationship, if any, in the four relevant palm pollen grains. The study was conducted by using Borassus and Phoenix antisera raised in rabbit. These antisera were used in rabbit IgG specific ELISA-inhibition and rocket immunoelectro-phoresis (RIE) assays for all four palm pollen extracts. In ELISA-inhibition, a distinct inhibition was obtained with comparable amount of soluble pollen protein. The RIE precipitin bands also revealed the presence of common antigenic components in the palm pollen. After isolation and purification, such common antigens may be useful in allergen immunotherapy in asthmatics.

Allergens↗

Clinical and bacteriological profile of neonatal infections in metropolitan city based medical college nursery.

Out of 6319 live born babies, 310 (4.9%) with high risk factors and with clinical suspicion of sepsis were studied for incidence pattern of neonatal infections. Incidence of neonatal infection was 2.56% in normal weight babies and 8.42% in low birth weight babies. Total culture positivity was 48.38%. Klebsiella was the most frequent offender, followed by Staph aureus and Staph epidermidis. Majority of klebsiella species were sensitive to ceftriaxone (86%), amikacin (84%) and ceftazidime (80%). Ceftriaxone was effective against Staph aureus (95%) and Staph epidermidis (95%). Overall fatality was 17.1%, distinctly higher in low birth weight group (22.06%) than normal weight group (6.18%).

Bacteria↗