Biomedical subjects
Publications and source records attributed to R Ghosh.
During 2004 and 2005, cholera was recorded in 15 states of India, with 7 outbreaks. The newly emerged Vibrio cholerae O1 Inaba had a different antibiogram and ribotype, different pulsotypes, and different mutations in the wbeT gene. Due to the absence of serogroup O139, the Inaba serotype may have acquired the potential to affect the population at large.
A cross sectional study on 387 Munda and 317 Pod adult males and females was conducted in the peri urban region of Kolkata city, India, to assess their health status. Chronic Energy Deficiency (CED) was calculated from the Body Mass Index (BMI) and seven anthropometric measurements were considered for this study. Significantly higher CED was observed among the Munda males and females. Higher percentage of CED was noticed among the females than the male Munda. Linear regression coefficient (b +/- standard error) of BMI on age and anthropometric measurements revealed significant decrease in BMI with increase in age among Munda males and females as well as Pod females. All other measurements more or less increased with increase in BMI. Thus, both intra- and inter-ethnic health differences were observed between and among the two socioeconomically backward populations. Higher gender differentials were found among the Munda and this needs an immediate attention of the government to overcome the health inequality at regional level.
Foot and mouth disease (FMD) is the major disease constraint on international trade in livestock and their products. In the state of West Bengal, India, 1,082 FMD outbreaks were reported in the 18 years from 1985 to 2002. Of the prevalent four serotypes, O type FMD virus accounted for the most outbreaks (67%), followed by Asia-1 virus type (15%) and A virus type (14%). Outbreaks of the type C FMD virus were least prevalent (4%), and no cases have been recorded since 1996. The study shows clearly that incidences were highest during the winter months and in the alluvial agro-climatic zones. The distribution and density of the FMD-susceptible population in different districts of the state also played major role in disease incidences. Due to the unrestricted movements of animals among different cattle markets, the disease was transmitted either by direct contact or by aerosols from infected to healthy animals. Cattle suffered most from the disease, accounting for about 95% of the cases. The monitoring of the disease documented in this study provides information about the endemicity of the disease that can help to formulate an effective strategy for an FMD control programme.
Antiosteoporotic activity of ormeloxifene, a multifunctional SERM, using inhibition in parathyroid hormone (PTH) induced resorption of 45Ca from prelabeled chick and rat fetal limb bones in chase cultures and modulation of certain biochemical markers of bone turnover and bone mineral density (BMD) in ovariectomized adult female rats, was investigated. Ormeloxifene concentration-dependently inhibited PTH-induced resorption of 45Ca from chick fetal femora with treated/control (T/C) ratio of 0.71, 0.32 and 0.20 at 50, 100 and 200 microM concentration, in comparison to 0.49, 0.53 and 0.95 in case of CDRI-85/287 (a pure antiestrogen), tamoxifen and ethynylestradiol (100 microM), respectively. Using rat fetal limb bones, ormeloxifene (100 microM) exhibited T/C ratio of 0.67, in comparison to 1.43 with PTH alone. Heat-killed bones exhibited negligible resorption (2.9%; T/C: 0.098) in response to PTH. In adult female rats, ormeloxifene (1.25 and 12.5 mg/kg per day) inhibited ovariectomy-induced increase in serum total and bone-specific alkaline phosphatase and osteocalcin and urine calcium/creatinine ratio to almost intact control level. Ovariectomy was accompanied by marked decrease in bone mineral density of isolated femur and tibia, being maximum in femur neck (28.3%; P < 0.01) and midshaft (23.7%; P < 0.01), but only marginal (6.7%; P > 0.05) in region proximal to tibio-fibular separation point. Decrease in BMD based on T-/Z-score, too, was >2.5 S.D. than mean value of normal young adult/age-matched females. This was prevented by ormeloxifene and the effect, though apparently more in females supplemented with higher dose of ormeloxifene, was not always significantly different and clear dose-response was not evident until BMD data was evaluated on T-/Z-score basis. The analysis also demonstrated much higher threshold level of tibia than femur and more so for their mid-shafts. Increase in BMD of isolated bones was also observed in ormeloxifene-treated intact females, without significantly altering biochemical markers of bone turnover or uterine weight. Findings suggest potential of ormeloxifene in management of post-menopausal osteoporosis and beneficial effect on BMD in women taking this SERM for contraception or any hormone-related clinical disorder.
Hybrid automata are an eminently suitable modelling framework for biological protein regulatory networks, as the protein concentration dynamics inside each biological cell are modelled using linear differential equations; inputs activate or deactivate these continuous dynamics through discrete switches, which themselves are controlled by protein concentrations reaching given thresholds. This paper proposes an iterative refinement algorithm for computing discrete abstractions of a class of hybrid automata with piecewise affine continuous dynamics and forced discrete transitions, defined completely in terms of symbolic variables and parameters. Furthermore, these discrete abstractions are utilised to compute symbolic parametric backward reachable sets from the equilibria of the hybrid automata, that are guaranteed to be exact or conservative under-approximations. The algorithm is then implemented using MATLAB and QEPCAD, to compute reachable sets for the biologically observed equilibria of the multiple cell Delta-Notch protein signalling automaton with symbolic parameters. The results are analysed to show that novel, non-intuitive, and biologically interesting properties can be deduced from the reachability computation, thus demonstrating the utility of the algorithm.
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The dynamics of an open quantum system exhibiting the quantum Brownian motion is analyzed when the coupling between the system and its environment is nonlinear, and the system and the reservoir are initially correlated. For couplings quadratic in the environment variables, the influence functional for the system is obtained perturbatively up to second order in the coupling constant, and then the propagator is explicitly evaluated when the particle is under the influence of a harmonic potential and an additional anharmonic potential, the so-called washboard potential. As an application of the propagator, the master equation and the Wigner equation are obtained for the quantum Brownian particle moving in a harmonic potential for the generalized correlated initial condition, and then for the specific case of the simplified "thermal" initial condition. The system is shown to obey the corresponding fluctuation-dissipation theorem.
1. Cryopreserved human hepatocytes were used to predict in vivo hepatic clearance (CL(hepatic)) from estimates of in vitro intrinsic clearance (CL' int). 2. (CL' int) was estimated for phenytoin, valproic acid, carbamazepine, theophylline, quinidine and procainamide after their addition to hepatocytes suspended either in human serum or in serum-free media. (CL' int)was estimated from in vitro concentration versus time data fitted to a monoexponential decay model. (CL' int) was estimated from concentrations measured at four time points and from just two-point measures, namely the initial concentration (C(0)) and the final concentration measurement (C(last)). 3. Predicted CL(hepatic) was within twofold of reported in vivo values of CL(hepatic) for all substrates. Moreover, predictions were not significantly different whether derived from hepatocytes suspended in serum or in serum-free medium. 4. Two-point estimates of (CL' int) were just as accurate in predicting CL(hepatic) as were multipoint estimates of (CL' int). 5. Although the data set was limited, the findings suggest that the measurement of the disappearance of xenobiotics from serum or serum-free media in which primary human hepatocytes have been suspended provides a physiologically relevant estimate of hepatic clearance that can be employed early in the drug development process to eliminate xenobiotics with unacceptable clearances.
Chinese hamster V79 cells were repeatedly exposed to a low dose of hydrogen peroxide (H2O2) over several weeks and then exposed to H2O2, cisplatin or ultraviolet (UV) light. Cell killing was examined by colony formation, following these treatments. It was seen that cells conditioned by multiple low doses of H2O2 showed resistance to killing in case of H2O2 and cisplatin but the sensitivity to UV light was same as the control cells. Apoptosis was also determined in these cells after the same treatments. UV light failed to induce apoptosis in both conditioned and in control cells, but in case of cells treated with H2O2 and with cisplatin, there was less apoptosis in the conditioned cells compared to the control cells. From our observation we can say that the enhanced survival of cells after treatment with H2O2 or cisplatin could be due to inhibition of apoptosis.
Abutilon mosaic geminivirus (AbMV) encodes two movement proteins, BV1 and BC1, which mediate the intra- and intercellular transport of viral DNA in plants cooperatively. It has been shown previously that singly expressed BC1, fused to green fluorescent protein (GFP), accumulates preferentially either at the cell periphery or around the nucleus in separate plant cells. To define the BC1 domains responsible for understanding the subcellular sorting, deletion mutants were fused to GFP and expressed transiently in epidermal cells of non-host (Allium cepa) as well as of host (Nicotiana benthamiana) plants with basically the same results in both species. BC1-mediated intracellular sorting was dependent on two protein domains, an "anchor domain" (amino acids 117 to 180) which is necessary and sufficient to fix GFP:BC1 at the cell periphery and the nuclear environment, and a "pilot domain" (amino acids 1 to 49) in the absence of which the fusion proteins were found at both sites in the same cell simultaneously.
A cross sectional study of the physical growth status was made on 655 Kamar children (341 boys and 314 girls), aged 5 to 18 years, in the Raipur district of Chhattisgarh. The study aimed to find out the growth pattern of the Kamar children, which is considered to be a primitive tribe of Chhattisgarh, India and was compared with another Indian tribe and the official data for all India (ICMR). Anthropometric measurements included height, weight, sitting height, biacromial diameter, biilliocrystal diameter, upper arm circumference, calf circumference and measurements of the triceps and subscapular skinfolds. All anthropometric measurements except skinfold thickness exhibit uniform increase with age in both sexes. However, when height and weight of the Kamar boys and girls were compared with the data for other tribes and for all India, the Kamar children (both boys and girls) indicated lower weight and height and the difference showed to be significant, for almost all ages. Kamar boys showed higher anthropometric values than girls in almost all measurements except in biilliocrystal diameter and in measured skinfolds. Poor socio-economic status of this primitive tribe may be one of the reasons for this poor growth pattern. However, in-depth study is necessary in order to arrive at any basic conclusions and to recommend any policy and interventions.
Membrane chromatography can overcome some of the problems associated with packed bed chromatography. In most membrane chromatographic studies reported so far, ion-exchange and affinity interactions have been utilised. In this paper the use of hydrophobic interactions for chromatographic separation is described. A polyvinylidene fluoride membrane was identified which could bind specific proteins in the presence of high ammonium sulphate concentration. The separation of CAMPATH-IG monoclonal antibody and bovine serum albumin using this membrane is discussed.
This article discusses a novel mode of operation for ultrafiltration processes termed Carrier Phase Ultrafiltration (CPUF). CPUF is based on a modification of dead-end ultrafiltration. Macromolecular fractionation using ultrafiltration is strongly influenced by operating and physicochemical parameters and requires precise "fine-tuning." CPUF facilitates the fractionation of high value biological macromolecules at optimised conditions and has several advantages over conventional modes of ultrafiltration. In this article the fractionation of two model proteins, lysozyme (MW 14100) and myoglobin (MW 17000), by CPUF using 25-kDa MWCO polysulfone membrane is discussed. Fractionation was carried out using two different CPUF modes, 1) pulse input CPUF and 2) step input CPUF. A high recovery of pure product was obtained in each case. These results are compared with those obtained from "conventional" ultrafiltration experiments.
Age-dependent changes in the effects of the alpha1-adrenoceptor agonist, phenylephrine were investigated on neurally evoked contractile responses and basal tone in smooth muscle strips from rat urinary bladder. Phenylephrine facilitated the neurogenic contractions in both neonatal and 7-month-old adult rats. However, phenylephrine increased the basal tone in adult but not neonatal rats. In adult rats, phenylephrine-induced facilitation of neurally evoked contractions occurred before and after the block of cholinergic contractions with 1 microM atropine. In adult rats, the phenylephrine facilitation was reduced at stimulation parameters (20 Hz, 80 shocks and maximal voltage) which activated muscarinic receptor mediated facilitation of acetylcholine release. The results indicate that pre-synaptic alpha1-adrenoceptors facilitate the release of both acetylcholine and the non-cholinergic non-adrenergic transmitter. In summary, alpha1-adrenoceptor-mediated facilitation is less expressed when muscarinic M1 receptor mediated facilitation is functioning; pre-junctional alpha1-adrenoceptors are present in the bladder of both neonatal and adult rats, whereas post-junctional alpha1-adrenoceptors are expressed only in older adult rats.
The enzyme ethoxyresorufin-o-deethylase (EROD) of the cytochrome P4501A family (CYP1A) in fish liver is increasingly being used as a molecular marker for qualitative and quantitative estimation of aquatic pollution throughout the world. The regulation and expression of this enzyme protein is very important from the toxicological point of view. The regulation of gene expression for this enzyme is mediated by the aromatic hydrocarbon receptor. In addition, cellular glutathione status influences expression of CYP1A. In this study, we explored the relationships among glutathione, EROD, and copper in cultured hepatocytes from Indian catfish. EROD activity in cultured hepatocytes was induced by carbofuran (CF), a widely used agricultural pesticide, and by beta-napthoflavone (BNF), a known inducer of CYP1A. Addition of copper into the culture media of hepatocytes inhibited EROD activity significantly. The activity of EROD elevated by CF and BNF was inhibited in hepatocytes pretreated with CF and BNF exposed to CuSO4. This effect was reflected in the glutathione status of the cells. The level of glutathione was increased by 3.4 and 3.0 times in hepatocytes treated with CF and BNF, respectively. These levels were inhibited in hepatocytes exposed to CuSO(4). Thus, copper interactions with glutathione may play a role in regulating EROD in hepatocytes.
Giardia lamblia, the ancient eukaryote does not have nucleolus but produces the fibrillarin protein that may be used for pre-rRNA processing. The nucleoli of eukaryotes contain complex population of small nucleolar RNAs, known as snoRNAs, several of which are required for rRNA processing. This report describes the full-length cloning of fibrillarin gene from Giardia lamblia, using RTPCR and the production of recombinant fibrillarin protein in Escherichia coli strain BL21 (DE3) as N-terminal His-tag protein. The condition for production of soluble protein was standardized. The expressed protein was purified by using Ni-chelation chromatography and used for functional studies. The small nuclear RNAs (snRNAs), RNA D, RNA J, and RNA H, containing box C, box D, and box C/D, respectively, of Giardia were also cloned by RTPCR. Antibody raised against the recombinant protein was used to identify the fibrillarin in giardial nuclear extract. The interaction of snRNAs with recombinant fibrillarin was followed using North-Western hybridization. Gel electrophoresis mobility shift assay demonstrated that bacterially expressed protein may participate in the in vitro interaction with RNA J, RNA H, and RNA D. Our results indicate that the recombinant fibrillarin by itself is able to bind and does not require the involvement of any other protein for this binding to the three snRNAs.
Among the 10 commonly used therapeutic agents investigated, concurrent oral administration of tetracycline (140 mg/kg) twice daily on Days 1-5 post-coitum (pc) interfered with the post-coital anti-implantation activity and almost completely abolished estrogen antagonistic activity of the single anti-implantation (1.5 mg/kg, orally) dose of dl-ormeloxifene administered on Day 1 pc, resulting in the occurrence of resorbed implantations in 50% of the females. However, no such interaction was evident when tetracycline was administered intramuscularly or when ormeloxifene was administered at twice its anti-implantation dose. There was no effect of ormeloxifene and/or tetracycline treatment on serum estradiol and progesterone levels, and all animals presented apparently normal corpora lutea. Ormeloxifene administered per se inhibited aminopyrine-N-demethylase (AD), glucose-6-phosphate dehydrogenase (G-6-PDH) and glutathione-S-transferase (GST) in the liver on the day of maximal endometrial receptivity, which was prevented by tetracycline co-administration. Aniline hydroxylase and AD were not detected in small intestine or uterus in vehicle control or any of the treatment groups. There was, however, no effect of ormeloxifene plus tetracycline treatment on serum total alkaline phosphatase activity. Findings suggest that interference with anti-implantation action of ormeloxifene by tetracycline might be due primarily to the almost complete abolition of its estrogen antagonistic activity at the uterine level, effected by decreased bioavailability of ormeloxifene and/or its active metabolite(s) by altered enterohepatic recirculation because of the effect on gut microflora. This might alternatively be related to an increased rate of its metabolism and elimination from the system via prevention of ormeloxifene-induced inhibition of hepatic AD, G-6-PDH, and GST, which, by effecting a decreased rate of metabolism, might be responsible for prolonged (approximately 120 h) duration of estrogen antagonistic/anti-implantation action of ormeloxifene in this species.
We investigated whether carbofuran (CF), a carbamate pesticide, at sub-lethal concentration had any adverse effects on reproductive function of the Indian catfish, Heteropneustes fossilis. 17beta-Estradiol content of serum and ovary of pre-spawning (P) and spawning (S) fish was reduced after sub-lethal concentration of carbofuran treatment (0.5-2 mg/ml, 30 days). After 30 days of CF treatment, the serum and ovarian vitellogenin levels of fish at the P stage were also reduced but remained unaltered in the S stage. The staining intensity of the pituitary gonadotrophs of the pre-spawning fish was significantly higher in CF-treated fish compared to controls suggesting the inability of the pituitary gonadotrophs to release gonadotropin following CF treatment. CF thus acts as an antiestrogenic, endocrine-disrupting agent in fish, possibly targeting the pituitary-gonad axis.