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Biomedical subjects

S Mahapatra

Publications and source records attributed to S Mahapatra.

At least 19 recordsLinked to original sources

Daily intake of aluminium by adult population of Mumbai, India.

Electrothermal atomic absorption spectrophotometry (ET-AAS) has been used for the determination of Al in environmental and food samples with a detection limit of 0.3 ng ml(-1). The reliability of estimation is assessed through the analysis of Standard Reference Materials (IAEA SRMs) of Soil-7, SL-3, SD-M2/TM and Hay (V-10). The results indicate that the average concentration of aluminium in air particulate samples is 5.3 microg m(-3). The daily intake of Al by the adult population of Mumbai is 6.4 mg day(-1).

Adult↗

Selenium levels in biological matrices in adult population of Mumbai, India.

Selenium (Se) levels in whole blood, serum, urine, muscle and saliva of Mumbai adults have been estimated by differential pulse cathodic stripping voltammetry (DPCSV); the detection limit of Se is 0.05 ng ml(-1). The reliability of estimation is further assessed through the analysis of Standard Reference Materials. The Se levels in whole blood (n = 35) and blood serum (n = 201) of the Mumbai adult population is 99.6 and 100 ng ml(-1); approximately 34.8% of the population have serum Se levels between 80 and 100 ng ml(-1). The blood serum levels of Se for Mumbai adults are comparable to those of whole blood. The mean concentration of Se in urine on a 24-h basis is 5.2 ng ml(-1). Muscle and saliva of the Mumbai adult population contain 195.4 ng g(-1) and 2 ng ml(-1) of Se, respectively. A good correlation between serum Se and dietary Se is observed with a correlation coefficient of 0.89.

Adult↗

Blood lead and its effect on Cd, Cu, Zn, Fe and hemoglobin levels of children.

The levels of heavy metals, such as Pb, Cd, Cu, Zn and Fe, in whole blood samples of Mumbai and Hyderabad children have been determined. In the present study, 576 blood samples of children (3-6 years old) were collected during 1996-1998 and analyzed for heavy metal contents by anodic stripping voltammetry and atomic absorption spectrometry. The geometric mean concentrations of Pb, Cd, Cu and Zn in blood of Mumbai children were 8.0, 0.10, 86.5 and 398.9 microg dl(-1), while those for Hyderabad children were 13.3, 0.13, 100.6 and 483.4 microg dl(-1) respectively. The geometric mean concentrations of Fe in the blood of Mumbai and Hyderabad children were 21.9 and 34.5 mg dl(-1), respectively. Of the Mumbai children, 61.8% were found to have a blood lead concentration lcss than 10 microg dl(-1), while the corresponding proportion of Hyderabad children was 27.4%. Higher concentrations of Pb in the children's blood were observed at high traffic areas in both cities. A decreasing trend in the hemoglobin content with increasing blood lead levels was observed for Mumbai children. A good negative correlation (-0.61) between the blood lead and iron has been observed for Mumbai children The hemoglobin content of 85% of the study population was found to vary between 8 and 14 g%.

Child↗

Daily intake of Se by adult population of Mumbai, India.

The atmospheric Se levels in Mumbai varied between 0.02 and 1.92 ng m(-3) with a mean concentration of 0.21 ng m(-3). The daily intake of Se by the adult population of Mumbai is 61.9 microg day(-1). Differential pulse cathodic stripping voltammetry (DP-CSV) has been used for the determination of Se in air particulate and food samples. The detection limit of Se using DPCSV was found to be 0.05 ng ml(-1). The reliability of estimation was further assessed through the analysis of standard reference materials (SRMs), tuna fish, animal blood, milk powder and fish tissue obtained from IAEA. The frequency distribution of dietary intake levels of selenium for Mumbai adults showed that approximately 61% of the studied population have a dietary intake of 30-90 microg day(-1). Ingestion was found to be the main route of Se exposure for Mumbai adults. The turn over time of Se through blood is 17 days.

Air Pollution↗

Purification, enzymatic characterization, and inhibition of the Z-farnesyl diphosphate synthase from Mycobacterium tuberculosis.

We have recently shown that open reading frame Rv1086 of the Mycobacterium tuberculosis H37Rv genome sequence encodes a unique isoprenyl diphosphate synthase. The product of this enzyme, omega,E,Z-farnesyl diphosphate, is an intermediate for the synthesis of decaprenyl phosphate, which has a central role in the biosynthesis of most features of the mycobacterial cell wall, including peptidoglycan, arabinan, linker unit galactan, and lipoarabinomannan. We have now purified Z-farnesyl diphosphate synthase to near homogeneity using a novel mycobacterial expression system. Z-Farnesyl diphosphate synthase catalyzed the addition of isopentenyl diphosphate to omega,E-geranyl diphosphate or omega,Z-neryl diphosphate yielding omega,E,Z-farnesyl diphosphate and omega,Z,Z-farnesyl diphosphate, respectively. The enzyme has an absolute requirement for a divalent cation, an optimal pH range of 7-8, and K(m) values of 124 micrometer for isopentenyl diphosphate, 38 micrometer for geranyl diphosphate, and 16 micrometer for neryl diphosphate. Inhibitors of the Z-farnesyl diphosphate synthase were designed and chemically synthesized as stable analogs of omega,E-geranyl diphosphate in which the labile diphosphate moiety was replaced with stable moieties. Studies with these compounds revealed that the active site of Z-farnesyl diphosphate synthase differs substantially from E-farnesyl diphosphate synthase from pig brain (Sus scrofa).

Alkyl and Aryl Transferases↗

Antimicrobial activity of new coumarin derivatives.

A preliminary exploration of coumarin analogs as novel antimicrobial agents was carried out to determine the basic features of the structure responsible for the observed biological activity. The substituents ester or carboxylic acid on the coumarin ring were needed to have potent inhibitory activity against both Gram-positive and Gram-negative bacteria. The presence of phenolic hydroxyl group and/or carboxylic acid was necessary to possess higher activity against Helicobacter pylori.

Anti-Bacterial Agents↗

Biosynthesis of the arabinogalactan-peptidoglycan complex of Mycobacterium tuberculosis.

The compositional complexity of the mycobacterial cell envelope differentiates Mycobacterium species from most other prokaryotes. Historically, research in this area has focused on the elucidation of the structure of the mycobacterial cell envelope with the result that the structures of the mycolic acid-arabinogalactan-peptidoglycan complex from M. tuberculosis are fairly well understood. However, the current impetus for studying M. tuberculosis and other pathogenic mycobacteria is the need to identify targets for the development of new drugs. Therefore, emphasis has been shifting to the study of cell envelope biosynthesis and the identification of enzymes that are essential to the viability of M. tuberculosis. The publication of the complete M. tuberculosis genome in 1998 has greatly aided these studies. To date, thirteen enzymes involved in the synthesis of the arabinogalactan-peptidoglycan complex of M. tuberculosis have been identified and at least partially characterized. Eleven of these enzymes were reported subsequent to the publication of the M. tuberculosis genome, a clear indication of the rapid evolution of knowledge stimulated by the sequencing of the genome. In this article we review the current understanding of M. tuberculosis arabinogalactan-peptidoglycan structure and biosynthesis.

Carbohydrate Conformation↗

Further biochemical characterization of Mycobacterium leprae laminin-binding proteins.

It has been demonstrated that the alpha2 chain of laminin-2 present on the surface of Schwann cells is involved in the process of attachment of Mycobacterium leprae to these cells. Searching for M. leprae laminin-binding molecules, in a previous study we isolated and characterized the cationic proteins histone-like protein (Hlp) and ribosomal proteins S4 and S5 as potential adhesins involved in M. leprae-Schwann cell interaction. Hlp was shown to bind alpha2-laminins and to greatly enhance the attachment of mycobacteria to ST88-14 Schwann cells. In the present study, we investigated the laminin-binding capacity of the ribosomal proteins S4 and S5. The genes coding for these proteins were PCR amplified and their recombinant products were shown to bind alpha2-laminins in overlay assays. However, when tested in ELISA-based assays and in adhesion assays with ST88-14 cells, in contrast to Hlp, S4 and S5 failed to bind laminin and act as adhesins. The laminin-binding property and adhesin capacity of two basic host-derived proteins were also tested, and only histones, but not cytochrome c, were able to increase bacterial attachment to ST88-14 cells. Our data suggest that the alanine/lysine-rich sequences shared by Hlp and eukaryotic H1 histones might be involved in the binding of these cationic proteins to laminin.

Animals↗

Characterization of derivatives of the high-molecular-mass penicillin-binding protein (PBP) 1 of Mycobacterium leprae.

Mycobacterium leprae has two high-molecular-mass multimodular penicillin-binding proteins (PBPs) of class A, termed PBP1 and PBP1* [Lepage, Dubois, Ghosh, Joris, Mahapatra, Kundu, Basu, Chakrabarti, Cole, Nguyen-Disteche and Ghuysen (1997) J. Bacteriol. 179, 4627-4630]. PBP1-Xaa-beta-lactamase fusions generated periplasmic beta-lactamase activity when Xaa (the amino acid of PBP1 at the fusion junction) was residue 314, 363, 407, 450 or 480. Truncation of the N-terminal part of the protein up to residue Leu-147 generated a penicillin-binding polypeptide which could still associate with the plasma membrane, whereas [DeltaM1-R314]PBP1 (PBP1 lacking residues Met-1 to Arg-314) failed to associate with the membrane, suggesting that the region between residues Leu-147 and Arg-314 harbours an additional plasma membrane association site for PBP1. Truncation of the C-terminus up to 42 residues downstream of the KTG (Lys-Thr-Gly) motif also generated a polypeptide that retained penicillin-binding activity. [DeltaM1-R314]PBP1 could be extracted from inclusion bodies and refolded under appropriate conditions to give a form capable of binding penicillin with the same efficiency as full-length PBP1. This is, to the best of our knowledge, the first report of a soluble derivative of a penicillin-resistant high-molecular-mass PBP of class A that is capable of binding penicillin. A chimaeric PBP in which the penicillin-binding (PB) module of PBP1 was fused at its N-terminal end with the non-penicillin-binding (n-PB) module of PBP1* retained pencillin-binding activity similar to that of PBP1, corroborating the finding that the n-PB module of PBP1 is dispensable for its penicillin-binding activity.

Bacterial Proteins↗

Solubilization Site of Organic Perfume Molecules in Sodium Dodecyl Sulfate Micelles: New Insights from Proton NMR Studies.

The site of incorporation of solubilizates in sodium dodecyl sulfate (SDS) micellar systems has been investigated by proton NMR spectroscopy. The solubilizate molecules chosen for the present study are phenol, 4-methylphenol, 4-allyl-2-methoxyphenol, anisole, 4-methylanisole, 4-propenylanisole, 1,8-cineole, and limonene. These molecules possess a wide variety of functional groups with different degrees of hydrophilic/hydrophobic character and are thereby solubilized at different micellar locations. Aromatic compounds, especially those having a phenolic-OH group, showed a large upfield shift of SDS methylene protons that are closely linked to the terminal sulfate groups. Additionally, in the case of phenolic compounds, the unresolved signals of the nine straight-chain bulk methylene protons of SDS are split into a broad doublet with uneven intensity. This splitting of methylene protons was found to be dependent on the concentration of the substrate. Based on these observations, probable solubilization sites and orientation of the substrate molecule within the micelles are discussed. Phenolic compounds, being the most hydrophilic among the present set, reside at the hydrophilic/hydrophobic boundary of micelle-water interface and thus influence the resonances of SDS protons the most. Aromatic methoxy and aliphatic compounds, being relatively more hydrophobic in nature, reside inside the micellar core and thereby result in smaller shifts. Copyright 2000 Academic Press.

Journal Article↗

Daily intake of manganese by the adult population of Mumbai.

The daily intake of manganese (Mn) estimated through air, water and duplicate dietary analysis is found to range from 0.67 to 4.99 mg with a mean value of 2.21 mg. Ingestion through food contributed to the predominant fraction of the intake. The turnover rate of Mn through blood is approximately 2 h, based on the mean concentration of Mn in blood of 1.54 microg l(-1). The average concentrations of Mn in water and air were approximately 1.42 microg l(-1) and 37 ng m(-3), respectively. The daily intake of Mn by the adult population of Mumbai is closer to the lower bound of the recommended limit of 2-5 mg. Electro Thermal Atomic Absorption Spectrophotometry (ET-AAS), has been used for the determination of Mn in a variety of environmental and human biological fluids. The detection limit of Mn for a volume injection of 20 microl is 2 pg absolute. The precision of the method is established by analyzing a synthetic mixture containing various elements in different quantities (0.5-10 ppm) and is found to be within +/- 8%. The reliability of estimation is further assessed through the analysis of Standard Reference Materials (SRMs) of soil, hay, milk powder and fish tissue obtained from IAEA.

Adult↗

Analysis of intracardiac electrograms showing monomorphic ventricular tachycardia in patients with implantable cardioverter-defibrillators.

Ventricular tachycardia (VT) initiation and its relation to various clinical factors was studied by reviewing intracardiac electrograms from patients with implantable cardioverter-defibrillators. Events were divided into (1) sudden onset without preceding ventricular premature complexes (VPCs), (2) extrasystolic onset with VPCs, or (3) paced, depending on the type and morphology of the last 5 beats before initiation of VT. Prematurity index, sinus rate, cycle length, and presence of short-long-short sequence for each episode was noted. A total of 268 episodes of VT among 52 patients were analyzed. Extrasystolic initiation was the most frequent pattern (177; 66%) followed by sudden onset (75; 28%) and paced (16; 6%). Among extrasystolic onset, 99 episodes (56%) were due to multiple VPCs and 149 episodes (84%) had different VPC morphology than the subsequent VT. Among pacing-induced VT, 13 of 16 episodes were due to inappropriate pacing due to undersensing of prior R waves. Sudden-onset episodes were slower (mean cycle length 383+/-97 ms) than extrasystolic (mean cycle length 336+/-88 ms, p = 0.002) and paced (mean cycle length 313+/-85 ms, p = 0.01) onset. Patients in the sudden-onset group had better left ventricular ejection fraction (33+/-15%) than the extrasystolic (29+/-11%, p<0.001) and paced (28+/-14%, p<0.01) groups. Extrasystolic onset with multiple, late coupled VPCs was the most common pattern of VT initiation and was associated with lower ejection fraction. Sudden-onset initiation was more common with better preserved systolic function.

Aged↗

Bacterial and host-derived cationic proteins bind alpha2-laminins and enhance Mycobacterium leprae attachment to human Schwann cells.

It has recently been demonstrated that laminin alpha2 chains present on the surface of Schwann cells are involved in the process of attachment of Mycobacterium leprae to these cells. In this study, a protein in the M. leprae cell wall that was found to be capable of binding alpha2-containing laminins (merosin) was isolated and characterized. The M. leprae laminin-binding protein was identified as a 21-kDa histone-like protein (Hlp), a highly conserved cationic protein present in other species of mycobacteria. The gene that encodes this protein was PCR amplified, cloned, and expressed, and the recombinant protein was shown to bind alpha2-laminins. More significantly, when added exogenously, Hlp was able to greatly enhance the attachment of mycobacteria to ST88-14 human Schwann cells. The capacity to bind alpha2-laminins and to enhance mycobacterial adherence to Schwann cells was also found in other cationic proteins such as host-derived histones. Moreover, mutation in the hlp gene was shown not to affect the capacity of mycobacteria to bind to ST88-14 cells, suggesting that alternative adhesins and/or pathways might be used by mycobacteria during the process of adherence to Schwann cells. The potential role of Hlp as a fortuitous virulence factor contributing to the pathogenesis of M. leprae-mediated nerve damage is discussed.

Adhesins, Bacterial↗

Comparison of the UDP-N-acetylmuramate:L-alanine ligase enzymes from Mycobacterium tuberculosis and Mycobacterium leprae.

In the peptidoglycan of Mycobacterium leprae, L-alanine of the side chain is replaced by glycine. When expressed in Escherichia coli, MurC (UDP-N-acetyl-muramate:L-alanine ligase) of M. leprae showed K(m) and V(max) for L-alanine and glycine similar to those of Mycobacterium tuberculosis MurC, suggesting that another explanation should be sought for the presence of glycine.

Alanine↗

The D arm of tRNATyr is necessary and sufficient for import into Leishmania mitochondria in vitro.

Transfer RNAs are selectively imported from the cytoplasm into mitochondria of kinetoplastid protozoa such as Leishmania . The specific structural features of tRNA which determine selectivity are largely unknown. Using an in organello system from Leishmania , the import signals on tRNATyrand on a synthetic transcript which binds to the same receptor, were studied by deletion and reconstruction analyses. In both cases, short oligoribonucleotides (minihelices) containing the sequence UGGYAGAG were imported with high efficiency in the presence of ATP. This motif is present in the D arm of tRNATyr, as well as in the majority of imported Leishmania tRNAs. Deletion of the D arm, or a point mutation in the conserved motif, reduces importability. The import signal coincides with the binding site for the mitochondrial receptor TAB. tRNAGln, which is not imported, forms non-productive, TAB-independent complexes with the mitochondrial surface. However, the observation that the imported:bound ratio of the D arm minihelix is higher than that of the entire molecule suggests that the post-binding translocation step is constrained in terms of size or structural flexibility. Kinetic studies of minihelix import indicate stepwise insertion of the molecule into import channels.

Animals↗

Study on body mass index, lipid profile and lipid peroxidation status in coronary artery disease.

Among the many known risk factors of coronary artery disease (CAD) obesity and hypercholesterolaemia are important ones. Whatever may be the risk factor, the basic pathology of CAD is deposition of altered lipids on the endothelium. One of such altered lipid is oxidatively modified low density lipoprotein (LDL). Lipid peroxidation has been assessed by several methods. Quantitation of malondialdehyde (MDA) by thiobarbituric acid (TBA) method is one of the commonly utilised method in several laboratories. In this study 40 cases of CAD were selected for evaluation. The body mass index (BMI), lipid profile and the level of lipid peroxidation (MDA) were measured. Seventeen cases (42.5%) had normal BMI (20-25), 20 cases (50%) were in the overweight range of BMI (26-30) and only 3 cases (7.5%) were in the obese group with a BMI more than 30. BMI correlated better with the level of total cholesterol (Tc), low density lipoprotein cholesterol (HDLc) and MDA. BMI did not show any correlation with triglyceride (Tg) or high density lipoprotein cholesterol (HDLc). MDA level correlated better with Tc, Tg levels and BMI, poorly correlated with LDLc and in inverse relationship was observed with HDLc.

Aged↗

Role of an RNA-binding protein in import of tRNA into Leishmania mitochondria.

Nuclear-encoded cytoplasmic tRNAs are imported into the mitochondria of kinetoplastid protozoa by an unknown mechanism. In a Leishmania in organello system, ATP-dependent import of a cloned, unspliced tRNATyr(GUA) transcript was demonstrated by protection from ribonuclease, whereas import of a tRNAGln(CUG) transcript was much less efficient. Specific binding of tRNATyr to two mitochondrial surface proteins of 15 and 22 kilodaltons was observed. Tubulin antisense-binding protein (TAB), the 15-kilodaton species, was purified to apparent homogeneity by RNA affinity chromatography. TAB forms stable complexes with the D stem-loop region of tRNATyr. Immunocytochemical and cell fractionation experiments, combined with limited proteolysis, suggested the association of TAB with the outer mitochondrial membrane. Importantly, anti-TAB antibody specifically inhibited binding as well as import of tRNATyr and of a synthetic structural homolog. These results support the role of TAB as a membrane-bound receptor or carrier for RNA import into Leishmania mitochondria.

Animals↗

Dual multimodular class A penicillin-binding proteins in Mycobacterium leprae.

The ponA gene of cosmid L222 of the Mycobacterium leprae genome library encodes a multimodular class A penicillin-binding protein (PBP), PBP1. The PBP, labelled with a polyhistidine sequence, has been produced in Escherichia coli, extracted from the membranes with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and purified by Ni2(+)-nitrilotriacetic acid-agarose chromatography. In contrast to the pon1-encoded class A PBP1, PBP1 undergoes denaturation at temperatures higher than 25 degrees C, it catalyzes acyl transfer reactions on properly structured thiolesters, and it binds penicillin with high affinity.

Acylation↗