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Biomedical subjects

S Bhattacharya

Publications and source records attributed to S Bhattacharya.

At least 91 records · Page 5Linked to original sources
PubMed2001

Y box-binding factor promotes eosinophil survival by stabilizing granulocyte-macrophage colony-stimulating factor mRNA.

Short-lived peripheral blood eosinophils are recruited to the lungs of asthmatics after allergen challenge, where they become long-lived effector cells central to disease pathophysiology. GM-CSF is an important cytokine which promotes eosinophil differentiation, function, and survival after transit into the lung. In human eosinophils, GM-CSF production is controlled by regulated mRNA stability mediated by the 3' untranslated region, AU-rich elements (ARE). We identified human Y box-binding factor 1 (YB-1) as a GM-CSF mRNA ARE-specific binding protein that is capable of enhancing GM-CSF-dependent survival of eosinophils. Using a transfection system that mimics GM-CSF metabolism in eosinophils, we have shown that transduced YB-1 stabilized GM-CSF mRNA in an ARE-dependent mechanism, causing increased GM-CSF production and enhanced in vitro survival. RNA EMSAs indicate that YB-1 interacts with the GM-CSF mRNA through its 3' untranslated region ARE. In addition, endogenous GM-CSF mRNA coimmunoprecipitates with endogenous YB-1 protein in activated eosinophils but not resting cells. Thus, we propose a model whereby activation of eosinophils leads to YB-1 binding to and stabilization of GM-CSF mRNA, ultimately resulting in GM-CSF release and prolonged eosinophil survival.

3' Untranslated Regions↗
PubMed2001

Insight to structural subsite recognition in plant thiol protease-inhibitor complexes : understanding the basis of differential inhibition and the role of water.

BACKGROUND: This work represents an extensive MD simulation / water-dynamics studies on a series of complexes of inhibitors (leupeptin, E-64, E-64-C, ZPACK) and plant cysteine proteases (actinidin, caricain, chymopapain, calotropin DI) of papain family to understand the various interactions, water binding mode, factors influencing it and the structural basis of differential inhibition. RESULTS: The tertiary structure of the enzyme-inhibitor complexes were built by visual interactive modeling and energy minimization followed by dynamic simulation of 120 ps in water environment. DASA study with and without the inhibitor revealed the potential subsite residues involved in inhibition. Though the interaction involving main chain atoms are similar, critical inspection of the complexes reveal significant differences in the side chain interactions in S2-P2 and S3-P3 pairs due to sequence differences in the equivalent positions of respective subsites leading to differential inhibition. CONCLUSION: The key finding of the study is a conserved site of a water molecule near oxyanion hole of the enzyme active site, which is found in all the modeled complexes and in most crystal structures of papain family either native or complexed. Conserved water molecules at the ligand binding sites of these homologous proteins suggest the structural importance of the water, which changes the conventional definition of chemical geometry of inhibitor binding domain, its shape and complimentarity. The water mediated recognition of inhibitor to enzyme subsites (Pn.H2O.Sn) of leupeptin acetyl oxygen to caricain, chymopapain and calotropinDI is an additional information and offer valuable insight to potent inhibitor design.

Amino Acid Chloromethyl Ketones↗
PubMed2001

Conventional in-vitro fertilisation versus intracytoplasmic sperm injection for the treatment of non-male-factor infertility: a randomised controlled trial.

BACKGROUND: Intracytoplasmic sperm injection (ICSI) is a more invasive option than conventional in-vitro fertilisation (IVF), which can be successful even when semen characteristics are poor. Reports of higher fertilisation rates after ICSI suggest that this technique may be better than the conventional method for all couples seeking IVF. We undertook a multicentre randomised controlled trial comparing clinical outcome after ICSI or traditional IVF in couples with non-male-factor infertility. METHODS: 415 eligible and consenting couples at four UK centres were randomly assigned IVF or ICSI (total 435 treatment cycles: IVF 224; ICSI 211). Usual clinical and laboratory protocols for the two treatment procedures were followed in each of four participating centres. The primary outcome was the implantation rate (number of gestation sacs per embryo replaced expressed as a percentage). Secondary outcomes were pregnancy and fertilisation rates associated with each treatment. Analyses were by intention to treat. FINDINGS: The implantation rate was higher in the IVF group than in the ICSI group (95/318 [30%] vs 72/325 [22%]; relative risk 1.35 [95% CI 1.04-1.76]). The pregnancy rate per cycle was also higher after IVF (72 [33%] vs 53 [26%]; 1.17 [0.97-1.35]). Mean associated laboratory time was significantly shorter with IVF than with ICSI (22.9 [SD 12.1] vs 74.0 [38.1] min; 95% CI for difference 45.6-56.6). INTERPRETATION: ICSI offers no advantage over IVF in terms of clinical outcome in cases of non-male-factor infertility. Our results support the current practice of reserving ICSI only for severe male-factor problems.

Adult↗
PubMed2001

Binding of a denatured heme protein and ATP to erythroid spectrin.

Spectrin is a large, worm-like cytoskeletal protein that is abundant in all cell types. The denatured heme enzyme, horseradish peroxidase showed significant decrease in the reactivation yield, after 30 min of refolding, in presence of increasing concentrations of spectrin from that in the absence. This indicated that spectrin could bind denatured HRP and inhibit their refolding. In presence of 1 mM ATP and 10 mM MgCl(2) the spectrin binding of denatured HRP is abolished. This activity of decreasing the reactivation yield was found to be ATP-dependent and the denatured enzyme after 30 min refolding in the presence of spectrin, pretreated with Mg/ATP, showed about 40% increase in the reactivation yield compared to the same in absence of spectrin. Fluorescence spectroscopic studies indicated binding of ATP to native spectrin showing concentration-dependent quenching of tryptophan fluorescence by ATP. The apparent dissociation constant of binding of ATP to spectrin was estimated to be 1.1 mM. A high affinity binding of spectrin with denatured HRP has been characterized (K(d) = 16 nM). Since these properties are similar to those of established molecular chaperone proteins, these data indicate that spectrin might have a chaperone-like function in erythrocytes.

Adenosine Triphosphate↗
PubMed2001

Structural and dynamic perturbations induced by heme binding in cytochrome b5.

The water-soluble domain of rat hepatic cytochrome b(5) undergoes marked structural changes upon heme removal. The solution structure of apocytochrome b(5) shows that the protein is partially folded in the absence of the heme group, exhibiting a stable module and a disordered heme-binding loop. The quality of the apoprotein structure in solution was improved with the use of heteronuclear NMR data. Backbone amide hydrogen exchange was studied to characterize cooperative units in the protein. It was found that this criterion distinguished the folded module from the heme-binding loop in the apoprotein, in contrast to the holoprotein. The osmolyte trimethylamine N-oxide (TMAO) did not affect the structure of the apoprotein in the disordered region. TMAO imparted a small stabilization consistent with an unfolded state effect correlating with the extent of buried surface area in the folded region of the native apoprotein. The failure of the osmolyte to cause large conformational shifts in the disordered loop supported the view that the specificity of the local sequence for the holoprotein fold was best developed with the stabilization of the native state through heme binding. To dissect the role of the heme prosthetic group in forcing the disordered region into the holoprotein conformation, the axial histidine belonging to the flexible loop (His63) was replaced with an alanine, and the structural properties of the protein with carbon-monoxide-ligated reduced iron were studied. The His63Ala substitution resulted in a protein with lower heme affinity but nevertheless capable of complete refolding. This indicated that the coordination bond was not necessary to establish the structural features of the holoprotein. In addition, the weak binding of the heme in this protein resulted in conformational shifts at a location distant from the binding site. The data suggested an uneven distribution of cooperative elements in the structure of the cytochrome.

Alanine↗
PubMed2001

A(1) or A(3) adenosine receptors induce late preconditioning against infarction in conscious rabbits by different mechanisms.

We investigated whether activation of A(1) or A(3) adenosine receptors (ARs) induces late preconditioning (PC) against infarction in conscious rabbits using the selective AR agonists 2-chloro-N(6)-cyclopentyladenosine (CCPA) and N(6)-3-iodobenzyladenosine-5'-N-methylcarboxamide (IB-MECA). In vitro radioligand binding and cAMP assays demonstrated CCPA to be approximately 200- to 400-fold selective for the rabbit A(1)AR and IB-MECA to be approximately 20-fold selective for the rabbit A(3)AR. We observed that (1) pretreatment of rabbits 24 hours earlier with CCPA (100 microgram/kg IV bolus) or IB-MECA (100 or 300 microgram/kg) resulted in an approximately 35% to 40% reduction in the size of the infarct induced by 30 minutes of coronary artery occlusion and 72 hours of reperfusion compared with vehicle-treated rabbits, whereas pretreatment with the selective A(2A)AR agonist CGS 21680 (100 microgram/kg) had no effect; (2) the delayed cardioprotective effect of CCPA, but not that of IB-MECA, was completely blocked by coadministration of the highly selective A(1)AR antagonist N-0861; (3) inhibition of nitric oxide synthase (NOS) with N(omega)-nitro-L-arginine during the 30-minute occlusion abrogated the infarct-sparing action of CCPA but not that of IB-MECA; and (4) inhibition of ATP-sensitive potassium (K(ATP)) channels with sodium 5-hydroxydecanoate during the 30-minute occlusion blocked the cardioprotective effects of both CCPA and IB-MECA. Taken together, these results indicate that activation of either A(1)ARs or A(3)ARs (but not A(2A)ARs) elicits delayed protection against infarction in conscious rabbits and that both A(1)AR- and A(3)AR-induced cardioprotection involves opening of K(ATP) channels. However, A(1)AR-induced late PC uses an NOS-dependent pathway whereas A(3)AR-induced late PC is mediated by an NOS-independent pathway.

Adenine↗
PubMed2001

Chemical control on the coordination mode of benzaldehyde semicarbazone ligands. synthesis, structure, and redox properties of ruthenium complexes.

Reaction of benzaldehyde semicarbazone (HL-R, where H is a dissociable proton and R is a substituent (R = OMe, Me, H, Cl, NO(2)) at the para position of the phenyl ring) with [Ru(PPh(3))(3)Cl(2)] and [Ru(PPh(3))(2)(CO2)Cl2] has afforded complexes of different types. When HL-NO(2) and [Ru(PPh(3))(3)Cl2] react in solution at ambient temperature, trans-[Ru(PPh(3))(2)(L-NO2Cl] is obtained. Its structure determination by X-ray crystallography shows that L-NO2 is coordinated as a tridentate C,N,O-donor ligand. When reaction between HL-NO2 and [Ru(PPh(3))(3)Cl2] is carried out in refluxing ethanol, a more stable cis isomer of [Ru(PPh(3))(2)(L-NO2)Cl] is obtained. The trans isomer can be converted to the cis isomer simply by providing appropriate thermal energy. Slow reaction of HL-R with [Ru(PPh(3))(2)(CO2)Cl2] in solution at ambient temperature yields 5-[Ru(PPh(3))(2)(L-R)(CO)Cl] complexes. A structure determination of 5-[Ru(PPh(3))(2)(L-NO2)(CO)Cl] shows that the semicarbazone ligand is coordinated as a bidentate N,O-donor, forming a five-membered chelate ring. When reaction between HL-R and [Ru(PPh(3))(2)(CO2Cl2] is carried out in refluxing ethanol, the 4-[Ru(PPh(3))(2)(L-R)(CO)Cl] complexes are obtained. A structure determination of 4-[Ru(PPh(3))(2)(L-NO2)(CO)Cl] shows that a semicarbazone ligand is bound to ruthenium as a bidentate N,O-donor, forming a four-membered chelate ring. All the complexes are diamagnetic (low-spin d(6), S = 0). The trans- and cis-[Ru(PPh(3))(2)(L-NO2)Cl] complexes undergo chemical transformation in solution. The 5- and 4-[Ru(PPh(3))(2)(L-R)(CO)Cl] complexes show sharp NMR signals and intense MLCT transitions in the visible region. Cyclic voltammetry of the 5-[Ru(PPh(3))(2)(L-R)(CO)Cl] and 4-[Ru(PPh(3))(2)(L-R)(CO)Cl] complexes show the Ru(II)-Ru(III) oxidation to be within 0.66-1.07 V. This oxidation potential is found to linearly correlate with the Hammett constant of the substituent R.

Journal Article↗
PubMed2001

Modeling the removal of VOC mixtures in biotrickling filters.

A mathematical model was derived for describing removal of mixed VOC vapors in biotrickling filters (BTFs). The model accounts for potential process rate limitation by the availability of oxygen as well as for potential kinetic interactions among pollutants during their biodegradation. Without using any fitted parameter, the model was found capable of predicting experimentally obtained removal rates of mono-chlorobenzene (m-CB) and ortho-dichlorobenzene (o-DCB) vapors. Experimental results reported here show that m-CB removal is better than that of o-DCB. The two compounds were known to be involved in a kinetic cross-inhibition interaction when degraded in suspended culture. However, model sensitivity studies showed that cross-inhibition does not affect BTF performance due to the low pollutant concentrations involved. For the same reason, the influence of oxygen on BTF performance was found to be minimal under the conditions tested. The model was found to predict experimentally obtained values with less than 10% error in the majority of cases. Computations with an earlier model describing VOC removal in conventional biofilters showed that, for the model mixture used in this study (m-CB/o-DCB), removal rates obtained with BTFs are one to more than two orders of magnitude higher than those obtained with conventional biofilters. This is attributed to the larger active specific biofilm surface area in BTFs, obtained through the creation of favorable growth conditions for the biomass, and better moisture control.

Air Pollutants↗
PubMed2001

Two coexisting vortex phases in the peak effect regime in a superconductor.

The critical current in the vortex phase of a type-II superconductor such as NbSe2 displays a striking anomaly in the vicinity of the superconductor-to-normal-metal transition. Instead of going to zero smoothly, it rebounds to a sharp and pronounced maximum, just before vanishing at the transition. This counter-intuitive phenomenon, known as the peak effect, has remained an unsolved problem for 40 years. Here we use a scanning a.c. Hall microscope to visualize the real-space distribution of the critical current in NbSe2. We show that in the peak-effect regime two distinct vortex-matter phases with intrinsically different pinning strengths coexist on a macroscopic scale. The composition of the two-phase mixture and the transformation of one phase into another are responsible for the history effects and anomalous voltage response commonly seen when external parameters such as temperature, magnetic field or transport current are varied. We argue that the observed phase coexistence is, in fact, the hallmark of a disorder-driven non-thermal phase transition.

Journal Article↗
PubMed2001

Molecular detection and characterization of circulating tumor cells and micrometastases in prostatic, urothelial, and renal cell carcinomas.

The detection and molecular characterization of circulating tumor cells (CTCs) and micrometastases in urinary tract and prostatic tumors may have important prognostic and therapeutic implications. In the last decade, numerous groups have attempted the detection of occult tumor cells in renal, prostatic, and urothelial carcinomas using the highly sensitive reverse-transcriptase polymerase chain reaction (RT-PCR). In prostatic carcinoma (PC), tissue-specific transcripts were detected with high specificity in the blood of patients with localized and advanced disease. PCR assays for PC detection were shown to be strong predictors of poorer outcome in some reports, while a lack of prognostic significance was found in other studies. There was a vast difference in the PCR positivity rates between various groups studying PC. This discrepancy could be due to variations in PCR methodology. In urothelial and renal cell carcinoma, the amount of research on the subject is still too limited. Currently, these assays for occult tumor cells are promising but are not yet ready to use in PC and urinary tract tumors. Because of the many limitations of PCR (e.g., false positives), many groups are developing new approaches for the detection of occult tumor cells. The most attractive technique involves immunomagnetic isolation of intact CTC and micrometastases prior to downstream analysis. The tumor-rich magnetic fraction can be subjected to RT-PCR, immunocytochemistry, and in situ hybridization. This will lead to better quantification and molecular characterization of these tumor cells.

Carcinoma, Renal Cell↗
PubMed2001

Reverse transcriptase polymerase chain reaction (RT-PCR) detection of melanoma-related transcripts in the peripheral blood and bone marrow of patients with malignant melanoma. What have we learned?

The detection of circulating tumor cells (CTC) and bone marrow micrometastases (BMM) by reverse transcriptase polymerase chain reaction (RT-PCR) may help predict recurrence and survival in malignant melanoma (MM). Since the appearance of the original article by Smith et al. in 1991 (Lancet 338:1227), several groups have attempted the detection of CTC and BMM in MM using RT-PCR for melanocytic specific markers, mainly tyrosinase mRNA. Most studies show that tyrosinase is not present in the PB and BM of control individuals without MM. The PCR positivity rates in MM are extremely variable, ranging from 0% to 100%. There was a correlation between RT-PCR and stage in some but not all of the studies. These disparate findings could in part be explained by differences in RNA extraction and blood separation techniques, to unrecognized contamination leading to false positive results, or differences in patient selection. Despite these discrepancies, we and others have shown that RT-PCR for tyrosinase mRNA in PB is able to predict overall survival (OS) and disease-free survival (DFS) in a statistically significant manner. In AJCC stage II-IV patients rendered surgically free of disease, we found that blood tyrosinase positivity was an independent predictor of OS and DFS. We also found that BM tyrosinase positivity is an independent predictor of DFS in the same group of patients. RT-PCR may help identify subgroups of patients at high risk for early relapse for more aggressive adjuvant therapy. Large prospective studies and interlaboratory quality assurance initiatives are necessary to confirm the accuracy and prognostic value of these RT-PCR assays.

Biomarkers, Tumor↗
PubMed2001

Insulin alone can lead to a withdrawal of meiotic arrest in the carp oocyte.

Meiotic arrest of oocyte in an Indian carp, Labeo rohita Ham. has been found for the first time to be withdrawn by insulin only. Addition of insulin to oocytes in vitro caused germinal vesicle breakdown (GVBD), one of the first visual markers to determine initiation of the final maturational process. Under the influence of insulin the germinal vesicle (GV) of the oocyte migrated towards the animal pole, reached the micropyle and then dissolved (GVBD). By using different concentrations of insulin i.e., 0.063, 0.63, 6.3 and 12.6 mM, optimum amount required was found to be 6.3 mM. Induction of GVBD by insulin could be blocked by cycloheximide (Chx), a translation inhibitor, while actinomycin D (AcD) had no effect suggesting non-involvement of transcriptional activity in this process. Addition of the maturation-inducing steroid 17alpha,20beta-dihydroxy-4-pregnen-3-one (DHP) stimulated (P<0.01) GVBD of carp oocytes and its combination with insulin showed an additive effect. Gonadotropin (GtH) caused GVBD but its effect was greatly augmented by insulin. Our results demonstrate that not only can insulin alone induce GVBD in carp oocytes, but it also augments the stimulatory effect of DHP or IGF-I or GtH on GVBD. This information will be important in hormonal manipulation during induced breeding of carp.

Animals↗
PubMed2001

Visual outcome in patients with isolated autosomal dominant congenital cataract.

OBJECTIVE: To determine the visual outcome and surgical complication rates of patients with isolated inherited congenital cataract. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Patients and their families were ascertained from the genetic eye clinic and outpatient databases of Moorfields Eye Hospital, London, and invited to participate in the study. Four hundred twenty-two individuals from 72 pedigrees with this form of autosomal dominant cataract underwent ophthalmologic assessment. MAIN OUTCOME MEASURES: Visual acuity and surgical complications (glaucoma, retinal detachment, amblyopia). RESULTS: In this study 49.4% of patients (46.8% of those operated) achieved a visual acuity (VA) of 20/40 or better, 35.9% (36.1% of those operated) a VA between 20/50 and 20/200, and 14.7% (17.1% of those operated) worse than 20/200. Opacities that were more diffuse or did not lie close to the visual axis were associated with a better prognosis for vision; 6.6% had glaucoma and 5.0% had retinal detachment develop. CONCLUSIONS: Patients with isolated inherited congenital cataract have a better visual and surgical outcome than those with coexisting ocular and systemic abnormalities. The improved prognosis is related in part to the lack of other developmental abnormalities of the eye, and, because inherited cataracts are often partial at birth, surgery may be delayed to later infancy and childhood when there is a lower incidence of surgical complications and refractive correction is easier. Certain inherited phenotypes (lamellar, pulverulent, polymorphic, coralliform, and cortical) also seem to have a better prognosis, and this should be borne in mind when counseling these families. A large number of the patients in this study underwent surgery many years previously, when surgical outcomes were less favorable, and thus the results of this study establish only a minimum acuity dataset for the purposes of counseling.

Adolescent↗
PubMed2001

Characterization of a retrotransposon-like element from Entamoeba histolytica.

The protozoan parasite Entamoeba histolytica is the causative agent of amoebiasis. The genome organization of this organism is not well understood. We had earlier reported the presence of a multicopy sequence, HMc, in E. histolytica. Subsequent analysis showed that HMc is a member of a retrotransposon family that we have named the E. histolytica retrotransposon-like element (EhRLE). Four other members of this family have been characterized. The EhRLE family is distributed across all chromosomes of the parasite. There are 140 copies, which show minor sequence variation with respect to one another (2--4% from the consensus sequence). From a sequence analysis of five members of the EhRLE family, the complete EhRLE unit is estimated to be 4086 bp in length. It has a 27-mer inverted repeat at its ends. A pairwise comparison with sequences in the database showed a highly significant match of a part of EhRLE with reverse transcriptases (RT), especially those encoded by non-long terminal repeat retrotransposons. There are stop codons in all the five EhRLEs, but a continuous open reading frame of 464 amino acids could be reconstructed by comparing the sequences of several EhRLEs. The reconstructed sequence showed a much better identity with RT as compared with any of the original EhRLE sequences. The non-pathogenic species, Entamoeba dispar, also contains this element, with 85% sequence identity with EhRLE. The data suggest that EhRLE may be a retrotransposon, but many of its members are probably nonfunctional due to the accumulation of mutations.

Amino Acid Sequence↗
PubMed2001

Life sciences flight hardware development for the International Space Station.

During the construction phase of the International Space Station (ISS), early flight opportunities have been identified (including designated Utilization Flights, UF) on which early science experiments may be performed. The focus of NASA's and other agencies' biological studies on the early flight opportunities is cell and molecular biology; with UF-1 scheduled to fly in fall 2001, followed by flights 8A and UF-3. Specific hardware is being developed to verify design concepts, e.g., the Avian Development Facility for incubation of small eggs and the Biomass Production System for plant cultivation. Other hardware concepts will utilize those early research opportunities onboard the ISS, e.g., an Incubator for sample cultivation, the European Modular Cultivation System for research with small plant systems, an Insect Habitat for support of insect species. Following the first Utilization Flights, additional equipment will be transported to the ISS to expand research opportunities and capabilities, e.g., a Cell Culture Unit, the Advanced Animal Habitat for rodents, an Aquatic Facility to support small fish and aquatic specimens, a Plant Research Unit for plant cultivation, and a specialized Egg Incubator for developmental biology studies. Host systems (Figure 1A, B: see text), e.g., a 2.5 m Centrifuge Rotor (g-levels from 0.01-g to 2-g) for direct comparisons between g and selectable g levels, the Life Sciences Glovebox for contained manipulations, and Habitat Holding Racks (Figure 1B: see text) will provide electrical power, communication links, and cooling to the habitats. Habitats will provide food, water, light, air and waste management as well as humidity and temperature control for a variety of research organisms. Operators on Earth and the crew on the ISS will be able to send commands to the laboratory equipment to monitor and control the environmental and experimental parameters inside specific habitats. Common laboratory equipment such as microscopes, cryo freezers, radiation dosimeters, and mass measurement devices are also currently in design stages by NASA and the ISS international partners.

Animals↗