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Biomedical subjects

A Singh

Publications and source records attributed to A Singh.

At least 523 records · Page 29Linked to original sources

Structure of polymerizable lipid bilayers IV. Mixtures of long chain diacetylenic and short chain saturated phosphatidylcholines and analogous asymmetric isomers.

Polymerization of 1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC8,9PC) is enhanced by addition of short-chain saturated phosphatidylcholines such as 1,2-dinonanoyl-sn-glycero-3- phosphocholine (DNPC). Because of well established constraints on the topochemical polymerisation process, we undertook structure-based experiments to determine the nature of this effect. Two hypotheses were tested: (a) that the DNPC crystalized the proximal (m) and disordered the distal (n) methylene segments of DC8,9PC, thus providing flexibility to accommodate the conformational change upon polymer formation, or (b) that the DNPC forced lateral displacement of DC8,9PC, which would then allow interdigitation of these segments with those of the opposing monolayer and potentially more crystalline alignment of the diacetylene. Low angle X-ray diffraction studies do not support the interdigiated chain model. However, these measurements also indicate that the two lipid species may be phase separated under many conditions. An analogous structure, 1-(tricosa-10,12-diynoyl)2-nonanoyl-sn-glycero-3- phosphocholine (C8,9NPC) did not polymerize, and low angle X-ray diffraction studies indicate that bilayers of this lipid were interdigitated such that the terminal methyl group of the tricosadiynoyl chain on each lipid in the bilayer was adjacent to the diacetylenic moiety of a lipid on the opposing monolayer. Implications of these findings pertinent to identifying significant factors in polymerization of diacetylenic phospholipid bilayers are discussed.

Fatty Acids↗

Acridine orange staining reaction as an index of physiological activity in Escherichia coli.

The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

Acridine Orange↗

Radionuclide assessment of blood-brain barrier disruption performed for chemotherapy of high grade malignant brain gliomas.

Compared to the conventional mode of chemotherapy of malignant brain gliomas following surgery and radiation therapy, chemotherapy after transient disruption of the blood-brain barrier coupled with intraarterial administration of methotrexate improved median survival from 12-14 to 22 months in our experience. Technetium-99m-DTPA brain scintigraphy played a unique and important role in the documentation of optimum blood-brain barrier disruption. Patients with excellent clinical outcome had significantly (P less than 0.0005) better blood-brain barrier disruption than patients with poor outcome. The results indicate that the clinical outcome is related to the degree of blood-brain barrier disruption.

Adolescent↗

Regulation of oestradiol 17 beta hydroxysteroid dehydrogenase in breast tissues: the role of growth factors.

Oestradiol 17 beta-hydroxysteroid dehydrogenase (E2DH) is present in normal and malignant breast tissues and regulates the interconversion of oestrone and the biologically active oestrogen, oestradiol. Studies we have previously carried out have indicated that concentrations of oestradiol and the conversion of oestrone to oestradiol are higher in breast tumours than in normal breast tissues. We are currently isolating and characterizing factors produced by breast tumours which are capable of stimulating E2DH (reductive) activity. The production of such factors by breast tumours, which stimulate the conversion of oestrone to oestradiol, would provide a favourable oestrogenic environment to promote tumour growth and may account for the increased concentrations of oestradiol in breast tumours.

17-Hydroxysteroid Dehydrogenases↗

Controlled delivery of diphtheria toxoid using biodegradable poly(D,L-lactide) microcapsules.

Diphtheria toxoid, which is an important vaccine in the expanded program of immunization (EPI) in the developing countries, was microencapsulated using poly(D,L,-lactide) of 49,000 molecular weight and the in-water drying technique. The microcapsules were subjected to an in vitro antigen release study using a sensitive enzyme-linked immunosorbent assay (ELISA) developed in the laboratory. Antibody titers in immunized Balb/C mice were also determined using direct ELISA. The antibody units in the immunized group till day 75 were quite comparable to those in the group receiving conventional three-dose injection of diphtheria toxoid with calcium phosphate as an adjuvant. SEM photographs of the microcapsules during in vitro degradation demonstrated the erosion kinetics of the polymer, leading to controlled release of the antigen.

Animals↗

An experimental evaluation of possible teratogenic potential in Boerhaavia diffusa in Albino rats.

The present study was undertaken to evaluate any possibility of teratogenic effects in Boerhaavia diffusa (Punarnava), a widely used herbal medicine for renal and urinary tract diseases by Ayurvedic physicians in India. The ethanolic extract of Boerhaavia diffusa (BDE) was administered daily in a dose of 250 mg/kg, body weight p.o., to pregnant albino female rats during the entire period of gestation. BDE was found to be devoid of any teratogenic effect as litter size and survival rate of foetuses were the same as for the normal control group and no foetal anomaly could be detected.

Animals↗

Biochemical indices of selected trace minerals in men: effect of stress.

Plasma zinc, iron, copper, and selenium and selected blood proteins were measured in 66 men before (BHW) and after (AHW) a 5-d period of sustained physical and psychological stress called Hell Week. Recovery blood samples were obtained from 26 men 7 d after Hell Week. Dietary intakes were determined BHW and during Hell Week; zinc, iron, copper, and selenium intakes during Hell Week averaged 23.6 +/- 3.4 mg/d, 35.4 +/- 3.9 mg/d, 3.0 +/- 0.5 mg/d, and 92.5 +/- 26.7 micrograms/d, respectively. C-reactive protein was detected in only five subjects BHW and in all subjects AHW. Zinc, iron, selenium, and albumin decreased by 33%, 44%, 12%, and 9%, respectively, whereas ferritin, ceruloplasmin, and creatine kinase concentrations increased AHW by 59%, 8%, and 266%, respectively. Haptoglobin concentrations increased 57% in 30 subjects but decreased 32% in 23 subjects AHW. The biochemical changes were transitory because protein (except ferritin) and mineral concentrations were similar to BHW values 7 d after Hell Week. Hell Week induced changes characteristic of an acute-phase response in physically active men.

Analysis of Variance↗

Characterization of Ribonuclease Activity of Three S-Allele-Associated Proteins of Petunia inflata.

Three S-allele-associated proteins (S-proteins) of Petunia inflata, a species with gametophytic self-incompatibility, were previously found to share sequence similarity with two fungal ribonucleases, RNase T(2) and RNase Rh. In this study, the S-proteins from P. inflata plants of S(1)S(2) and S(2)S(3) genotypes were purified to homogeneity by gel filtration and cation-exchange chromatography, and their enzymatic properties were characterized. The three S-proteins (S(1), S(2), and S(3)), with pairwise sequence identity ranging from 73.1 to 80.5%, were similar in most of the enzymatic properties characterized. The ribonuclease activity had a pH optimum of 7.0 and a temperature optimum of 50 degrees C. Diethylpyrocarbonate at 1 millimolar almost completely abolished the ribonuclease activity; cupric sulfate and zinc sulfate at 1 millimolar reduced the ribonuclease activity of the three S-proteins by 50 to 75%. EDTA and RNasin had no inhibitory effect. All three S-proteins hydrolyzed polycytidylic acid preferentially, but varied in their nucleolytic activity toward polyadenylic acid and polyuridylic acid.

Journal Article↗

The influence of fractions of abnormal erythrocytes on aggregation.

Erythrocytes with abnormal shapes and deformability reduce cell aggregation and hence sedimentation. It is not known what influence small fractions of these abnormal cells have on the sedimentation behaviour of normal cells. We have used three different methods to alter erythrocyte properties (glutaraldehyde, heat treatment, and metabolic depletion). The admixture of such cells in increasing fractions to normal cells affected the sedimentation behaviour in three different ways. Heat-treated erythrocytes decreased the sedimentation proportionally. Small fractions of glutaraldehyde-treated cells increased the sedimentation, which was explained by a pro-aggregatory change of surface properties by glutaraldehyde. With 50% or more glutaraldehyde-treated cells sedimentation was inhibited, which was due to excessive formation of aggregates occupying the entire tube. The presence of small fractions of echinocytes produced by metabolic depletion decreased cell sedimentation disproportionally, which was explained by an inability of spiculated cells to form aggregates with normal erythrocytes. These results indicate that erythrocyte sedimentation is affected by fraction of abnormal cells in a complex way. Many diseases are characterized by fractions of abnormal erythrocytes. Our results may, therefore, contribute to a better understanding of these conditions.

Adenosine Triphosphate↗

Obesity and polycystic ovary syndrome.

Our studies show that obese women with polycystic ovary syndrome are more likely to have hirsutism and menstrual disturbances than are lean women with PCOS. The most obvious biochemical differences between obese and lean women with PCOS is that SHBG concentrations are much lower in women with obesity. The SHBG levels are inversely related to insulin, and insulin has been shown to have a direct inhibitory action on SHBG secretion. Other factors, however, may contribute to the mechanism of the increased prevalence of hirsutism and anovulation in obese women with PCOS, such as a direct effect of insulin or increased activity of 5 alpha-reductase in peripheral tissues. Finally we have been able to show that weight reduction of more than 5% is associated with an improved biochemical profile and, importantly, with restoration of fertility.

Endocrine Glands↗

Blood rheology in acute mountain sickness and high-altitude pulmonary edema.

The role of blood rheology in the pathogenesis of acute mountain sickness and high-altitude pulmonary edema was investigated. Twenty-three volunteers, 12 with a history of high-altitude pulmonary edema, were studied at low altitude (490 m) and at 2 h and 18 h after arrival at 4,559 m. Eight subjects remained healthy, seven developed acute mountain sickness, and eight developed high-altitude pulmonary edema. Hematocrit, whole blood viscosity, plasma viscosity, erythrocyte aggregation, and erythrocyte deformability (filtration) were measured. Plasma viscosity and erythrocyte deformability remained unaffected. The hematocrit level was lower 2 h after the arrival at high altitude and higher after 18 h compared with low altitude. The whole blood viscosity changed accordingly. The erythrocyte aggregation was about doubled 18 h after the arrival compared with low-altitude values, which reflects the acute phase reaction. There were, however, no significant differences in any rheological parameters between healthy individuals and subjects with acute mountain sickness or high-altitude pulmonary edema, either before or during the illness. We conclude that rheological abnormalities can be excluded as an initiating event in the development of acute mountain sickness and high-altitude pulmonary edema.

Acute Disease↗

Pseudomonas aeruginosa: a probe to detail change with age in corneal receptor(s).

This in vitro study used Pseudomonas aeruginosa as a probe to examine the change with age of the host receptor(s) facilitating bacterial adherence to cornea in the immature eye and to pinpoint in time, when the host receptor(s) was no longer able to be recognized by the bacterium. Scanning EM was used to quantitate adherent organisms at the corneal surface of mice of select ages (5-37 postnatal days, P, and 28 months) at 15, 30 and 60 min following bacterial application. Organisms were most numerous at the surface of the 5-P eye at all time periods. In 9-21-P mice, no bacterial binding was observed until 30 min, and bacterial adherence modestly increased at 60 min after bacterial application. In the 37-P and in aged 28-month-old mice, no binding to cornea was observed until 60 min after bacterial application. Statistical analysis of these data revealed significant differences in the mean number of bacteria binding to cornea between age groups.

Aging↗

Insulin-like growth factor type I and insulin-like growth factor type II stimulate oestradiol-17 beta hydroxysteroid dehydrogenase (reductive) activity in breast cancer cells.

Oestradiol-17 beta hydroxysteroid dehydrogenase (E2DH) is present in normal and malignant breast tissues and also in cultured breast cancer cells. It can act in a reductive direction to convert oestrone to the biologically active oestrogen, oestradiol, or in an oxidative direction to metabolize oestradiol to oestrone and may therefore have a crucial role in regulating breast tissue concentrations of oestradiol. Insulin-like growth factor-type I (IGF-I) and IGF-II are both mitogens for breast cancer cells. In this study we have examined the effect of these growth factors on the reductive and oxidative activities of E2DH in MCF-7 (receptor positive) and MDA-MB-231 (receptor negative) breast cancer cells. Both IGF-I (80 ng/ml) and IGF-II (80 ng/ml) significantly stimulated E2DH reductive activity (up to 138%) in MCF-7 cells but had no effect on oxidative activity. Addition of IGF-II (100 ng/ml) to MDA-MB-231 cells resulted in a small but statistically significant (p less than 0.05) increase in E2DH reductive activity (18%) but in these cells reductive activity is 25-70 times lower than oxidative activity. If IGF-I and IGF-II act to stimulate E2DH reductive activity in breast tumours then such a mechanism could account for the increased concentrations of oestradiol detected in breast tumours.

Breast Neoplasms↗

Analysis of adhesion, piliation, protease production and ocular infectivity of several P. aeruginosa strains.

The role of bacterial piliation and protease production in Pseudomonas aeruginosa adhesion to the injured corneal epithelial surface and subsequent infectivity was examined using several bacterial strains, including three that were hyperpiliated. To initiate this study, bacteria were examined by transmission EM to confirm their piliation characteristics. The PAK strain, like pseudomonas ATCC 19660, possessed about 1-4 polar pili. The mutant PAK/PR11 lacked pili while PAK/PR1, DB2, a mutant of PAO1, and PA1244, a wild-type clinical isolate, were hyperpiliated. Ocular infectivity of these bacterial strains and mutants was examined macroscopically and histopathologically in mice and these data compared to the well-characterized ocular disease response of a murine model of infection with pseudomonas ATCC 19660. The PAK strain was infective, but less virulent than strain 19660 by both macroscopic grading and histopathological analysis of infected eyes. Infectivity of the PR11 mutant was similar to the PAK parent strain, while PR1, DB2 and 1244, all hyperpiliated, were not infective. To explore the hypothesis that hyperpiliated bacteria bound less well to cornea and thus failed to induce corneal disease, in vitro quantitative studies of bacterial adhesion were done using an ocular organ culture model. The PR1 hyperpiliated mutant bound significantly less well to cornea than the PAK parent strain, PR11 mutant or pseudomonas 19660, while DB2 and 1244 binding did not differ significantly from 19660 or PAK. Examination of protease production, another factor which may influence adhesion, revealed that only 19660 and DB2 produced detectable protease. This study provides evidence that non-piliated, non-protease producing strains such as PAK/PR11 possess alternate virulence mechanisms to facilitate binding to and infectivity of corneal tissue.

Animals↗