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Biomedical subjects

S Mishra

Publications and source records attributed to S Mishra.

At least 19 recordsLinked to original sources

High-resolution holocene environmental changes in the thar desert, northwestern india

Sediments from Lunkaransar dry lake in northwestern India reveal regional water table and lake level fluctuations over decades to centuries during the Holocene that are attributed to changes in the southwestern Indian monsoon rains. The lake levels were very shallow and fluctuated often in the early Holocene and then rose abruptly around 6300 carbon-14 years before the present (14C yr B.P.). The lake completely desiccated around 4800 (14)C yr B.P. The end of this 1500-year wet period coincided with a period of intense dune destabilization. The major Harrapan-Indus civilization began and flourished in this region 1000 years after desiccation of the lake during arid climate and was not synchronous with the lacustral phase.

Journal Article

Reduced sarcoplasmic reticulum Ca(2+)-uptake and expression of phospholamban in left ventricular myocardium of dogs with heart failure.

The purpose of this study was to examine the sarcoplasmic reticulum (SR) Ca(2+)-uptake and the expression of phospholamban (PLB) and Ca(2+)-ATPase (CAA) in left ventricular (LV) and right ventricular (RV) myocardium of 6 normal (NL) dogs and 6 dogs with chronic heart failure (HF). In addition, gene expression of PLB and CAA was also examined in LV myocardium of NL and HF dogs. HF (LV ejection fraction 23+/-2%) was produced by multiple sequential intracoronary microembolizations. Oxalate-dependent Ca(2+)-uptake was measured in isolated membrane vesicles. Using specific dog myocardial monoclonal antibody, the expression of CAA, PLB and calsequestrin (CSQ) were measured in sodium dodecyl sulfate extract prepared from LV and RV tissue. Steady-state mRNA levels were determined by Northern hybridization using specific cDNA clones of PLB, CAA, CSQ, and glyceraldehyde-3-phosphate dehydrogenase (GADPH), a house keeping gene. SR Ca(2+)-uptake of NL and HF dogs increased with increasing Ca(2+)concentrations and reached a plateau at 3 microm in both LV and RV. Total capacity (134+/-9 v 224+/-10 nmol(45)Ca/mg protein/10 min, P<0.05) and maximal velocity (15+/-2 v 2 nmol(45)Ca/mg protein/min, P<0.05) of the SR to sequester Ca(2+)was significantly lower in LV myocardium of HF dogs compared to NL, whereas the Hill coefficient and the affinity of the Ca(2+)-pump for Ca(2+)were unchanged. LV tissue levels of the PLB and CAA, normalized to noncollagen protein or to CSQ and the PLB and CAA mRNA levels, normalized to CSQ or GADPH mRNA, were also significantly lower in HF dogs compared to NL. In RV myocardial tissue, no significant differences in total capacity of SR to sequester Ca(2+), maximal velocity of SR Ca(2+)-uptake, the affinity and Hill Coefficient of the Ca(2+)-pump for Ca(2+), or tissue levels of PLB and CAA were observed between NL dogs compared to HF dogs. We conclude that SR Ca(2+)-uptake and SR PLB and CAA protein and gene expression levels are reduced in LV myocardium of dogs with chronic HF. These abnormalities can lead to Ca(2+)-overload and subsequent global LV dysfunction.

Animals

Direct fluorometry of phase-extracted tryptamine-based fast quantitative assay of L-tryptophan decarboxylase from Catharanthus roseus leaf.

An assay for the enzyme L-tryptophan decarboxylase (TDC; EC 4.1.1.28) is described. It is based on direct fluorometry of the enzymatic reaction product (tryptamine) selectively recovered in ethyl acetate from the reaction mixture. Catalytically formed tryptamine from tryptophan in the incubation mixture is selectively (free from tryptophan) physically separated as ethyl acetate solution under basic (pH > or = 11) conditions and subjected to direct fluorescence measurement in the organic solvent using a spectrofluorometer with excitation and emission wavelengths of 280 and 350 nm, respectively. Tryptamine production rate was quantitated from the luminescence response curve of tryptamine drawn under similar extraction and measurement conditions. Luminescence calibration curves were drawn for tryptamine in aqueous (water or buffer system) as well as in organic solvent as recovered from the varied aqueous solution conditions including those similar to the enzyme incubation mixture. The luminescence calibration graphs were linear for at least 0.5 to 10 microM tryptamine. The examination of interassay variations and the comparative magnitude of fluorescence response allowed to infer that a satisfactory and sufficient sample luminescence response was retained under the varied conditions including those akin to the enzymatic assay mixture, allowing adaptation of the fluorometry for the TDC activity quantitation. The assay was found to follow the proportionality principle of product formation with respect to catalytic reaction time as well as protein concentration in the assay mixture using Catharanthus roseus leaf crude homogenate as well as the enzyme preparation at different states of purity. The rate of tryptamine formation under the catalytic conditions was linear for at least 1 h at 30 degrees C. Though the assay has been demonstrated to use the C. roseus leaf as the enzyme source, it should be equally applicable to other plant and nonplant sources. The merits and precautions of the protocol have been discussed.

Alkalies

A novel type of Ca2+ channel in U-87 MG cells activated by anti-galactocerebroside.

Antibody to galactocerebroside (GalC) evokes a Ca2+ response in cultured glioma U-87 MG cells. The rise in intracellular calcium [Ca2+]i occurs largely due to the influx of Ca2+ through a plasma membrane channel, though the release of Ca2+ from intracellular stores also contributes. We characterized the channel activated by anti-GalC. The channel activity was transient and the inactivation appeared to be Ca2+ dependent. The channel was impermeant to monovalent ions Na+ and K+ and also to Mn2+. Ni2+ and Co2+ neither permeate through the channel nor inhibit the Ca2+ influx. In contrast Cd2+ the most potent inorganic blocker of Ca2+ channels permeated through this channel. The Ca2+ influx was inhibited by verapamil with IC50 of 65 +/- 8 microM. The Ca2+ influx as well as the intracellular release were markedly inhibited by neomycin sulfate and phorbol dibutyrate, suggesting that the Ca2+ influx may be mediated by IP3 (1). Depletion of intracellular Ca2+ stores by thapsigargin was followed by Ca2+ influx. This represents the capacitative Ca2+ entry pathway and is distinct from the channel activated by anti -GalC.

Calcium

Correlation between presence of viable bacteria and presence of endotoxin in middle-ear effusions.

The presence of endotoxin (detected by the Limulus amebocyte lysate assay) was compared to the presence of viable Haemophilus influenzae and Moraxella catarrhalis (detected by PCR) in 106 middle-ear effusions from pediatric patients with chronic otitis media. Endotoxin was found in 81 of the 106 specimens. Of these 81 specimens, 66 (81.5%) also tested positive for one or both of the gram-negative bacteria H. influenzae and M. catarrhalis. The data suggest that viable gram-negative bacteria, detectable by PCR but often undetectable by culture, may be the source of endotoxin in middle-ear effusions.

Adolescent

Research-related burnout among faculty in higher education.

We investigated the effects of intrinsic motivation to do research and perceived lack of rewards contingent on doing research on burnout or disenchantment from research. Findings, based on a survey of 328 faculty at a major university, indicate that these two variables account for 74% and 81% of the variance in scores on a scale relating to burnout among 260 tenured and 68 untenured professors, respectively. The effect of perceived lack of rewards contingent on doing research on burnout was moderated by the tenure status.

Attitude

Acinetobacter sepsis in newborns.

OBJECTIVE: To evaluate the clinico-epidemiological profile of Acinetobacter sepsis in neonates. DESIGN: Retrospective study. SETTING: Level II Neonatal Care Unit. SUBJECTS: 79 neonates with blood culture positive for Acinetobacter. METHODS: Relevant information was collected on a predesigned proforma from the case records and analyzed for clinical and epidemiological characteristics. RESULTS: The incidence of Acinetobacter septicemia was 11.1/1000 live births. Fifty-five babies were hospital born, 24 were outborn. Out of these, 64.6% babies were born at term and 40.5% had a birth weight of 2500 g or more. A cluster of 53 cases was seen between May and September 1995. In cases with early onset sepsis (onset < 7 days of postnatal age), difficulty in breathing (n = 54), chest retraction (n = 35) and refusal to feed (n = 46) were seen more commonly as compared to late onset sepsis (p < 0.05). Complications observed included meningitis, bleeding manifestations and necrotising enterocolitis in three, six and five babies, respectively. The organism was sensitive to ciprofloxacin (96.2%), amikacin (92.4%) and gentamicin (87.3%). A response rate of 52.4% was observed with Ciprofloxacin in babies not responding to cefotaxime and amikacin combination. The overall mortality was 13.9%. CONCLUSION: Nosocomial Acinetobacter sepsis may affect fullterm, appropriate for gestational age babies. Clinical presentation is indistinguishable from Gram negative septicemia. Life threatening complications can also occur. Ciprofloxacin may prove to be useful drug in resistant cases.

Acinetobacter Infections

Expression and characterization of Pichia etchellsii beta-glucosidase in Escherichia coli.

The beta-glucosidase enzyme is important as the terminal enzyme involved in hydrolysis of cellobiose and short-chain cellodextrins generated during enzymatic cellulose degradation. Under controlled reaction conditions the enzyme also displays cello-oligosaccharide synthesizing ability (based on either the thermodynamic or kinetic approach). We present here the purification of the enzyme beta-glucosidase (BGL) of Pichia etchellsii from recombinant pBG55 Escherichia coli clone. The kinetic parameters, substrate specificity and oligosaccharide synthesizing ability of the purified enzyme are also reported. The purified 200-kDa protein (tetramer of 50 kDa) was identified as a broad-substrate-specificity enzyme exhibiting increased temperature and glucose tolerance compared to the native yeast enzyme. Temperature directed substrate specificity for aryl beta,1-4 linkage, and beta (1-2), beta(1-4), beta(1-6) and beta(2-1) linkages in various natural disaccharides was observed. Glycosylation of the enzyme was found to be unimportant for enzyme activity. With both cellobiose and glucose, oligosaccharide synthesis was detected. The implications of this information with regard to cellulose hydrolysis and oligosaccharide synthesis are discussed.

Blotting, Southern

Concurrent balloon dilatation of mitral and tricuspid stenosis during pregnancy using an Inoue balloon.

Percutaneous balloon dilatation of combined mitral and tricuspid stenoses was performed successfully in a 22-year-old woman during the 23rd week of pregnancy, using an Inoue balloon catheter. Following valvuloplasty, mean transmitral gradients decreased from 36 to 5 mmHg and the mean transtricuspid gradients from 12 to 3 mmHg. The procedure was uncomplicated and well tolerated, leading to the vaginal delivery of a healthy newborn at term. The fluoroscopy time required was 84 s for mitral and 18 s for tricuspid valvuloplasty. The present report highlights the advantage of the Inoue balloon catheter for multivalvular dilatation during pregnancy with minimum fluoroscopy time.

Adult

Proportion and pattern of dystrophin gene deletions in north Indian Duchenne and Becker muscular dystrophy patients.

Population-based variations in frequency and distribution of dystrophin gene deletions have been recognized in Duchenne/Becker (DMD/BMD) muscular dystrophy patients. In the present study, DNA samples from 121 unrelated DMD/BMD patients from North India were analyzed for deletional studies with multiplex PCR and Southern hybridization. A total of 88 (73%) patients showed intragenic deletions in the dystrophin gene. The observed proportion of gene deletions is relatively high, particularly compared with that of Asian counterparts. However, the distribution of breakpoints across the gene does not show significant variations.

Dystrophin

Physiologic and pathophysiologic relevance of T-type calcium-ion channels: potential indications for T-type calcium antagonists.

The family of voltage-gated calcium-ion (Ca2+) channels is critical in the role of transmembrane signaling of excitable cells throughout the body. Within the cardiovascular system, two types of Ca2+ channels have been identified: the L-type channel and the T-type channel. These two types of Ca2+ channels have distinct electrophysiologic identities, and although the roles of the T-type Ca2+ channels have not been firmly established, there are many reasons for believing that the roles of the T-type and L-type Ca2+ channels are distinct. T-type Ca2+ channels have the appropriate characteristics to generate pacemaker activity in the sinoatrial node. In vascular smooth muscle, they appear to be involved in maintenance of coronary and peripheral vasomotor tone and control of vascular growth and remodeling. Characterization of the T-type Ca2+ channels will be facilitated by the availability of mibefradil, a novel calcium antagonist that selectively blocks T-type Ca2+ channels. Mibefradil is associated with a reduction in heart rate but not with negative inotropic effects or neurohormonal stimulation. It is thought that the unique pharmacologic effects of mibefradil are related to blockade of T-type Ca2+ channels, and it is hypothesized that this action will have a positive impact on cardiovascular morbidity and mortality via cardioprotective and renoprotective effects. However, much work needs to be done to fully test this hypothesis.

Angina Pectoris

Purification and characterization of diacylglycerol acyltransferase from the lipid body fraction of an oleaginous fungus.

Diacylglycerol acyltransferase (DGAT) [EC 2.3.1.20] was purified to apparent homogeneity from the lipid body fraction of an oleaginous fungus, Mortierella ramanniana var. angulispora. The enzyme was solubilized from the lipid body fraction with 0.1% Triton X-100, and purified by subsequent column chromatography on Yellow 86 agarose, Superdex-200, Heparin-Sepharose, second Superdex-200, and second Yellow 86 agarose. The enzyme activity was finally enriched 4,802-fold over that of the starting 1,500X g supernatant. The apparent molecular mass of the enzyme was 53 kDa on SDS polyacrylamide gel electrophoresis. The purified enzyme did not exhibit glycerol-3-phosphate acyltransferase, lysophosphatidic acid acyltransferase, lipase, transacylase, or acyl-CoA hydrolase activities, although 2-monoolein was acylated with about a half of the enzyme activity toward 1,2-diolein. The purified DGAT depended on exogenous sn-1,2-diolein and oleoyl-CoA, with the highest activity at about 200 and 20 microM, respectively. Purified DGAT utilized a broad range of molecular species of both diacylglycerol and acyl-CoA as substrates. The highest activity was observed with sn-1,2-diolein and lauroyl-CoA. Anionic phospholipids such as phosphatidic acid (PA) activated the purified enzyme, as found for the Triton X-100 extract. Sphingosine dose-dependently inhibited DGAT activity activated by PA and basal activity without PA. These results provide a basis for further studies on the molecular mechanism of triacylglycerol biosynthesis and lipid body formation, in which DGAT plays an important role.

Acyltransferases

Genotype-phenotype correlation in Duchenne/Becker muscular dystrophy patients seen at Lucknow.

The molecular basis of two allelic forms of muscular dystrophy, Duchenne (DMD) and Becker (BMD), has been explained by frame shift hypothesis. In order to test this hypothesis, deletional mutations in 59 patients confirmed to have DMD and 11 BMD patients were analysed using multiplex polymerase chain reaction and Southern hybridization with dystrophin cDNA probes. Translational reading frame of the dystrophin gene was derived from 'Border type' analysis of exons flanking the intragenic deletions. The correlation between genotype (reading frame) and phenotype (clinical severity) showed higher number of DMD patients (approximately 20%) deviating from the frame shift hypothesis. The patients who deviated had deletions at the central hot spot region of the dystrophin gene. The presence of these deviations in a large number of DMD patients highlights the difficulties in predicting the clinical progression of the disease based only on DNA profile.

Gene Deletion

Regulatory role of cortisone and dietary carbohydrate energy in prepubertal rat mammary gland growth and development.

Role of cortisone and regulated carbohydrate energy in mammary gland growth, based on studies of morphometric and certain growth and energy related biochemical parameters has been evaluated in the adrenalectomized prepubertal female rat. Results showed improvement or restoration of mammary features like fat pad weight, area, duct system and sprouting of end and lateral buds by cortisone treatment from adrenalectomy induced suppressed state. Similarly the hormone rectified the gland nucleic acids, glycogen and rate of tissue oxidation of glucose levels, reduced under adrenalectomy. Adrenalectomy caused loss of appetite (33.5% less than normal intake). This amount of diet restriction to normal intact did not affect the gland morphology but reduced nucleic acids, protein, glycogen and glucose oxidation rate. Provision of dextrose mixed diet to the adrenalectomized rats showed variable improvements of these biochemical parameters except the gland protein level in association with increase in appetite. However, cortisone therapy under provision of dextrose supply, restored gland protein level and increased further in nucleic acids, glycogen and glucose oxidation rate. The morphological growth parameters with dextrose and cortisone also showed further improvements. Results suggested that glucocorticoid was essential mammary growth factor during prepubertal ages. The hormone appeared to operate via tissue metabolism and stimulating energy intake through appetite stimulation.

Adrenalectomy