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Biomedical subjects

S Ali

Publications and source records attributed to S Ali.

At least 199 records · Page 11Linked to original sources
PubMed1996

Paraquat induced DNA damage by reactive oxygen species.

The redox cycling contact herbicide paraquat (PQ) causes oxidative damage to pulmonary tissue. PQ is reduced enzymatically to PQ radical in lung where it reacts with molecular oxygen, generating reactive oxygen species (ROS). ROS damage various macromolecules including DNA. However, the ability of paraquat to mediate DNA damage is unknown. In this study, Bam H1 site (5'-GGATCC-3') on pBR322 DNA was chosen as the target sequence for a study of the PQ-mediated DNA damage. The incubation of PQ with plasmid DNA in the presence of freshly prepared rat lung microsomes and NADPH resulted in damage to the restriction site. The PQ-treated DNA was not digested with the endonuclease reflected by the digestion pattern of DNA on agarose gels. The effect was dependent on the dose of PQ. The PQ-mediated damage to DNA was comparable to DNA damage caused by ROS generated through the xanthine-xanthine oxidase system. The results of the present study suggest that ROS generated by PQ in vitro under aerobic conditions may lead to a modification of the restriction site on DNA.

DNA Damage↗
PubMed1996

The heart of the senior oarsman: an echocardiographic evaluation.

We evaluated left ventricular mass and function in 15 oarsmen aged 78 (65-82) yr (median and range) and in 15 sedentary males aged 72 (65-81) yr by 2-D and M-mode echocardiography and cycle ergometry. The weekly time spent exercising among the oarsmen was 6 (2-18) h and two of the oarsmen were former national and international champions. The two groups of subjects had similar weight, height, and resting blood pressure. The oarsmen reached a maximal work rate of 142 (117-174) vs 113 (75-150) W for the sedentary group (P < 0.01). The internal diameters of the left ventricle were not significantly different, but the septum and posterior wall thicknesses were larger in the oarsmen (11 (8-20) vs 9 (7-11) mm, and 9 (8-13) vs 8 (7-19) mm, respectively, P < 0.023). Thus, the left ventricular mass index of the oarsmen was 19% larger (127 (101-284) vs 103 (74-134) g.m-2, P < 0.01). Also, the systolic function appeared to be superior in the oarsmen as the fractional shortening was 0.45 (0.28-0.55) vs 0.36 (0.18-0.49) in the controls (P < 0.05). In conclusion, we found that long-term rowing in the senior oarsman is associated with enlarged myocardial wall thickness, a normal systolic function, and a high work capacity.

Aged↗
PubMed1996

Genomic distribution of 5' TTCCA 3' repeat motif and its diagnostic potential in human Y-chromosome-related anomalies.

We have studied genomic distribution of a simple repeat motif 5' TTCCA 3' derived from the DYZ1 fraction of the human Y chromosome employing restriction fragment length polymorphism and in situ hybridization techniques. This has led us to develop a synthetic DNA based genetic marker specific to human genome. Randomly selected human genomic DNA from both sexes, digested with a total of 16 restriction enzymes, OAT20Y probe comprising four repeat units of 5'TTCCA 3' motif failed to reveal fragment length polymorphisms. In contrast, with most of the enzymes, several multilocus monomorphic bands and with a few enzymes, smeary signals were detected. In situ hybridization of the OAT20Y probe with human chromosomes revealed grains on the long arm of the Y chromosome, whereas the X-chromosome and autosomes showed random distribution of the grains without any preferential labeling in the centromeric or telomeric regions. The OAT20Y probe uncovers a 3.4 kb isomorphic band exclusively in the human male DNA digested with Hae III enzyme. Using the OAT20Y probe, we have detected the presence of Y chromosome in mosaic cell populations of Turner's patients with dysgenetic gonads and high levels of LS/FSH. The presence of Y chromosome in these patients has been associated with an increased risk of gonadoblastoma. The OAT20Y probe offers sensitivity and accuracy for the detection of Y-chromosome-bearing cells in a mosaic cell population and, consequently, help in better management of the patients.

Blotting, Southern↗
PubMed1996

Simultaneous measurement of cyclopentyladenosine-induced contraction and intracellular calcium in bronchial rings from allergic rabbits and it's antagonism.

We reported previously that adenosine is a specific contractile agonist in the asthmatic airways of allergic rabbits; cyclopentyladenosine (CPA) was the most potent bronchoconstrictor in this model. The aim of the present investigation was to examine the contracting effect of CPA, an A1 adenosine receptor agonist, in relation to the role of intracellular calcium ([Ca++]i) in airway smooth muscle of allergic rabbits in the presence and absence of extracellular calcium (Ca++). The effects of adenosine receptor antagonists theophylline (xanthine) and CGS-15943 (nonxanthine) were also evaluated on these responses. CPA (10(-9)-10(-4) M) produced a dose-dependent contraction of tertiary airway smooth muscle of allergic rabbits. An increase in tension of airway smooth muscle was accompanied by a quantitative increase in [Ca++]i in the presence of extracellular Ca++. CGS-21680, an A2 agonist, produced only marginal changes in force and [Ca++]i compared with CPA. The CPA-induced contraction as well as changes in [Ca++]i were significantly inhibited by both antagonists at a concentration of 10(-7) M. CGS-15943 and theophylline changed the EC50 values for the force from 1.1 x 10(-7) M to 2.3 x 10(-6) M and 1.0 x 10(-6) M, respectively. In Ca(++)-free medium, CPA (10(-4) M) induced only a 15 to 20% contraction compared with Ca(++)-containing medium. The changes in [Ca++]i were also reduced accordingly. CGS-15943 significantly inhibited the CPA-induced tension and changes in [Ca++]i whereas theophylline failed to inhibit these responses. 8-Cyclopentyl-1,3-dipropylxanthine, an A1-specific and potent antagonist, also did not inhibit the CPA-induced force and [Ca++]i in a separate set of experiments. The tertiary airway smooth muscle rings from age-matched normal rabbits did not respond to CPA and there was no detectable change in [Ca++]i. These data suggest that the asthmatic airway smooth muscle contraction has both extracellular Ca(++)-dependent and -independent components and the Ca(++)-independent component is xanthine insensitive.

Adenosine↗
PubMed1996

Differential scanning calorimetry study of the influence of phospholipid analogs with a carbonyl-terminated sn-2 chain on the interdigitated phases formed by 1-stearoyl-2-capryl-sn-glycero-3-phosphatidylcholine (C18:C10-PC).

Synthetic glycerophosphocholines with highly asymmetric chain lengths form interdigitated bilayers in the gel phase. In nature, phospholipids with one hydrocarbon chain approximately twice as long as the other can arise from the autooxidation of unsaturated linkages in the acyl chain, and thus the oxidation products would contain a carbonyl group at the chain terminus. In this study, we have investigated the thermotropic behavior of bilayers prepared from mixtures of the well-studied, mixed-chain phospholipid, 1-stearoyl-2-capryl-sn-glycero-3-phosphocholine (C18:C10-PC, 1), with synthetic 1-stearoyl-2-acyl-sn-glycero-3-phosphocholines in which the sn-2 chain is approximately one-half the length of the sn-1 chain and contains a C==O group near the omega terminus. Phase diagrams of binary mixtures of 1 with a chain-terminal ketone-PC analog (2) or with a chain-terminal ester-PC analog (3) in excess water exhibited gel-phase immiscibility over a wide compositional range, but miscibility in the liquid-crystalline phase. However, 1 was completely miscible with C18:C10:1 delta 10-PC (compound 4), which bears a chain-terminal carbon-carbon double bond, in both the gel and liquid-crystalline phases. The calorimetric data suggest that phosphatidylcholines (PC) with carbonyl-terminated chains, which can be produced by autooxidation of naturally abundant 1-saturated-2-unsaturated phospholipids such as 1-stearoyl-2-oleoyl-PC, may not form the normal triple-chain mixed interdigitated structure characteristic of hydrocarbon-terminated PCs in gel-phase bilayers.

Calorimetry, Differential Scanning↗
PubMed1995

Adapter function of protein-tyrosine phosphatase 1D in insulin receptor/insulin receptor substrate-1 interaction.

Insulin signal transduction involves the multisite docking protein insulin receptor substrate-1 (IRS-1) and a number of Src homology-2 (SH2) domain factors, including p85/p110 phosphatidylinositol 3-kinase, p110 GTPase-activating protein, and the phosphotyrosine-specific phosphatase PTP1D. In transfected baby hamster kidney cells, Rat1 fibroblasts, and normal IM9 lymphoblasts, PTP1D directly binds activated insulin receptor. This interaction is mediated by catalytic domain-proximal SH2 determinants of the phosphatase and phosphotyrosine 1146 of the activated insulin receptor. While the receptor and the phosphatase do not serve as substrates for each other, their interaction promotes IRS-1 binding to the receptor, indicating that PTP1D functions as an adapter for insulin receptor and IRS-1. The formation of a multiprotein signaling complex involving the insulin receptor, PTP1D, and IRS-1 enhances cellular glucose uptake, a critical process in the physiological action of insulin.

Amino Acid Sequence↗
PubMed1995

Protein-tyrosine phosphatase 1D modulates its own state of tyrosine phosphorylation.

The insulin receptor-mediated signal transduction pathway involves insulin receptor substrate 1 and a variety of proteins containing Src homology-2 (SH2) domains, such as phosphatidylinositol 3-kinase, Grb2, and protein-tyrosine phosphatase 1D (PTP1D). Upon insulin stimulation of baby hamster kidney cells overexpressing the IR, the catalytically inactive mutant of PTP1D, C463A, becomes tyrosine-phosphorylated and coprecipitates with Grb2. Tyrosine phosphorylation of this mutant is significantly reduced when wild type PTP1D is coexpressed. Substitution of tyrosine residues 546 and 584 with phenylalanine abrogates tyrosine phosphorylation of the catalytically inactive mutant and abolishes its interaction with Grb2.

Animals↗
PubMed1995

Proline-rich sequence-mediated Jak2 association to the prolactin receptor is required but not sufficient for signal transduction.

The prolactin receptor (PRLR) belongs to the superfamily of cytokine/growth hormone/prolactin receptors. Members of this family do not contain a tyrosine kinase domain but are associated with cytoplasmic kinases of the Jak family. Here, we examine different mutants of the PRLR with respect to their ability to associate and activate the kinase Jak2 and the transcription factor Stat1. Moreover, using a biological assay system we are able to correlate these activities with activation of prolactin-responsive gene transcription. Our results indicate that interaction between Jak2 and PRLR requires a proline-rich sequence in the membrane proximal region of the receptor, which is conserved among the different members of the cytokine receptor superfamily. We also show that association of Jak2 with the receptor is sufficient for activation of the kinase as well as the transcription factor Stat1. Moreover, our findings indicate that association of PRLR with Jak2 is necessary but not sufficient for the transmission of a lactogenic signal. We have identified two other cytoplasmic regions of the PRLR that are required for activation of transcription. These two regions are located between boxes 1 and 2 and are in the carboxyl-terminal tail of the receptor. These sites probably involve specific interactions with other effector molecules.

Amino Acid Sequence↗
PubMed1995

A single phosphotyrosine residue of the prolactin receptor is responsible for activation of gene transcription.

Members of the cytokine/growth hormone/prolactin (PRL) receptor superfamily are associated with cytoplasmic tyrosine kinases of the Jak family. For the PRL receptor (PRLR), after PRL stimulation, both the kinase Jak2 and the receptor undergo tyrosine phosphorylation. To assess the role of tyrosine phosphorylation of the PRLR in signal transduction, several mutant forms of the PRLR in which various tyrosine residues were changed to phenylalanine were constructed and their functional properties were investigated. We identified a single tyrosine residue located at the C terminus of the PRLR to be necessary for in vivo activation of PRL-responsive gene transcription. This clearly indicates that a phosphotyrosine residue in the cytoplasmic domain of a member of the cytokine/growth hormone/PRL receptor superfamily is directly involved in signal transduction.

Caseins↗
PubMed1995

Characterization of the amino-terminal transcriptional activation function of the human estrogen receptor in animal and yeast cells.

We have previously reported that the transcriptional activation function AF-1, located in the A/B region of the human estrogen receptor, exhibits cell-type and promoter context specificity in both animal cells and yeast. To further characterize AF-1, we have constructed a number of deletion mutants spanning the A/B region in the context of either the whole human estrogen receptor or the A/B region linked to the GAL4 DNA binding domain, and tested their transcriptional activity in chicken embryo fibroblasts and in yeast cells, two cell types in which AF-1 efficiently activates transcription on its own. Additionally, we utilized HeLa cells in which AF-1 is poorly active but can synergize with the transcriptional activation function AF-2 located in the hormone binding domain. We show that in animal cells the "independent" activity of AF-1 is embodied in a rather hydrophobic proline-rich 99-amino acid activating domain (amino acids 51-149), whereas amino acids 51-93 and 102-149 can independently synergize with AF-2. Interestingly, in yeast, three discrete activating domains (amino acids 1-62, 80-113, and 118-149) are almost as active on their own as the whole A/B region, indicating that multiple activating domains can operate independently in yeast. Our study also demonstrates that, within the context of the whole human estrogen receptor, the same AF-1 activating domains are "induced" by either estradiol or 4-hydroxytamoxifen.

Animals↗
PubMed1995

Characterization of the role of the stimulatory magnesium of Escherichia coli porphobilinogen synthase.

The synthesis of tetrapyrroles is essential to all phyla. Porphobilinogen synthase (PBGS) is a zinc metalloenzyme that catalyzes the formation of porphobilinogen, the monopyrrole precursor of all biological tetrapyrroles. The enzyme from various organisms shows considerable sequence conservation, suggesting a common fold, quaternary structure, and catalytic mechanism. Escherichia coli and plant PBGS are activated by magnesium, a property that is absent from mammalian PBGS. This stimulatory Mg(II) is called Mgc. Mgc is not required for activity and is distinct from the two zinc ions (ZnA and ZnB) common to mammalian and E. coli PBGS (PBGSE.coli). For PBGSE.coli, both the Km for the substrate 5-aminolevulinic acid (ALA) and the Vmax are altered by the presence of Mgc; Mg(II) causes the Km to drop from approximately 3 to 0.30 mM and the maximum specific activity to increase from 23 to 50 mumol h-1 mg-1. Mgc also causes the saturating concentration of the required Zn(II) to decrease from 0.1 mM to 10 microM. Maximal activation by Mg(II) occurs at 0.5 mM; thus, in E. coli the Mgc site is probably saturated under physiological conditions. Mn(II) is a good substitute for Mgc, giving a comparable increase in catalytic activity. Consequently, Mn(II) has been used as an EPR active probe of the Mgc binding site. Mn(II) binds at a stoichiometry of eight ions per enzyme octamer. The X- and Q-band EPR spectra reflect a single type of binding site with rhombic symmetry and are consistent with oxygen and/or nitrogen ligands. The addition of unlabeled or 1-13C-labeled ALA does not significantly affect the Mn(II) EPR spectra.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1995

Patterns of punitiveness in women with eating disorders.

The level and direction of hostility in patients with bulimia nervosa, anorexia nervosa and a comparison group were measured using the the Hostility and Direction of Hostility Questionnaire. A semistructured interview developed by Harris, Brown, and Bifulco (Psychological Medicine, 16, 641-659, 1986) was used to assess childhood care to examine whether a link exists between childhood exposure to aggression or parental neglect and adult hostility. Patients with eating disorders had significantly higher hostility levels and were significantly more intropunitive than the comparison group. Patients with bulimia nervosa were significantly more intropunitive than the comparison group. Patients with bulimia nervosa were significantly more hostile than patients with anorexia nervosa. Anorexia nervosa patients were more likely to direct hostility inwardly, rather than outwardly, when compared with bulimia nervosa patients. Impulsivity was associated with extrapunitiveness whereas intropunitiveness was associated with depression. Although some measures of poor childhood care correlated with adult hostility levels no clear pattern emerged.

Adolescent↗
PubMed1995

Superoxide radicals mediate the biochemical effects of methylenedioxymethamphetamine (MDMA): evidence from using CuZn-superoxide dismutase transgenic mice.

The subacute and long-term biochemical effects of methylenedioxymethamphetamine (MDMA) were assessed in homozygous and heterozygous transgenic (Tg) mice that carry the complete sequence of the human copper-zinc (CuZn) superoxide dismutase (SOD) gene. Non-transgenic (Non-Tg) mice showed significant decreased in striatal dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) levels both at 24 h and at 2 weeks after a single injection of MDMA (50 mg/kg). Heterozygous SOD-Tg mice showed DA depletion only at the 24 h time point. In contrast, homozygous SOD-Tg mice show no DA or DOPAC depletion at either the 24 h or at the 2 week time points. Moreover, three injections of MDMA (50 mg/kg) given 24 h apart also caused marked reduction of striatal DA and DOPAC in Non-Tg mice when these substances were measured 2 weeks after the last MDMA injection. That injection schedule also caused small decreases in DA levels in the heterozygous animals but no changes in the homozygous mice; DOPAC levels were not affected in the heterozygous nor in the homozygous SOD-Tg mice. Furthermore, the multiple injection schedule caused significant decreases in DA and DOPAC in female Non-Tg mice but not in the two strains of transgenic mice. Neither the single dose nor the multiple dose schedule of MDMA injections affected striatal serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels in any of the three strains of mice. These results support previous observations that MDMA-induced biochemical effects are observed in the DA systems of mice, whereas these effects are seen in the 5-HT systems of rats. The present observations also document for the first time a role for the production of superoxide radicals in these effects of MDMA. These mice are an important tool for dissecting pathways involved in drug-induced neurotoxicity.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

The diagnostic value of exercise echocardiography in ischemic heart disease in relation to quantitative coronary arteriography.

The aim of the study was to assess the diagnostic value of bicycle exercise echocardiography using quantitative coronary arteriography as a reference. Exercise echocardiography was performed in 70 consecutive patients referred for coronary angiography. Digital loops were obtained at rest, peak, and immediately after exercise in the standard views (parasternal long and short axis, apical two and four chamber views). Wall motion analysis was made on the basis of the 16 segment model, scoring each segment from 3 (hyperkinesia) to -1 (hypokinesia). Exercise echocardiography was considered positive when wall motion in at least one segment decreased at least one score from rest to peak or post exercise. Cinefilms were evaluated using automated quantitative coronary arteriography software. Transstenotic pressure gradients were calculated based on flow assumptions at the maximal stenosis flow reserve. Pressure losses > 30 mmHg and quantitatively measured percent diameter stenosis of > 50% were considered clinically significant. Stenoses in the equivocal range of 40-69% were subjected to separate analysis. Exercise echocardiography was superior to exercise-induced ST-segment depression in the diagnosis of coronary artery disease. In the overall sample of 70 patients, the sensitivity of exercise echocardiography against percent diameter stenosis was 84%, against pressure gradient 86%. The specificity against these two parameters was 86% and 84%, respectively. When analysing the subgroup of 40-69% stenoses (N = 14), sensitivity of exercise echocardiography against percent diameter stenosis was 67%, against pressure gradient 88%. The specificity against these two parameters was 100% and 84%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Studies on the role of glycosylation in the origin of the electrophoretic variants for rat corticosteroid-binding globulin.

The glycoprotein corticosteroid-binding globulin (CBG) migrates as doublet bands in PAGE and SDS-PAGE, and as numerous bands in isoelectric focusing (IEF). This study deals with the origin of this heterogeneity. Desialation of rat CBG with neuraminidase does not abolish the doublet in either PAGE or SDS-PAGE, indicating that the doublet does not arise as a result of differences in sialic acid residues. Treatment of the separated upper and lower variants of native CBG with N-glycosidase F (PNGase-F) shows a differential pattern of deglycosylation over time indicating either differences in the number, type, or location of sugars attached to each of the variants. Rate of deglycosylation is quicker and more extensive for the upper variant when compared to the lower variant. PNGase-F treatment of 1% SDS-denatured CBG does not abolish the CBG doublet seen in SDS-PAGE, indicating that there is variation in the protein moiety. Sugars could not be detected on PNGase-F treated CBG using either wheat germ aglutinin horse radish peroxidase conjugate, concavilin-A HRP conjugate, or the digoxigenin glycan detection system. While the results clearly show differences in glycosylation between the CBG variants, differences in the protein moiety may also occur to give rise to the heterogeneity seen in CBG. The latter is supported by the fact that desialated CBG migrates as two bands in IEF. Migration in IEF is based solely on charge, and since only sialic acid residues are charged in N-linked glycosylation, any heterogeneity seen for the desialated glycoprotein must reside within the protein moiety itself. The presence of O-glycosylation containing an N-acetylgalactosamine with a beta 1-3 linkage to galactose could not be demonstrated using O-glycosidase. N-terminal blockage could not account for the variation, as both the upper and lower variants were able to be sequenced resulting in identical sequences for the first 13 amino acids. The data presented supports the hypothesis that the differences in the sugar as well as the protein moiety are responsible for the heterogeneity seen for CBG.

Amidohydrolases↗
PubMed1995

Hepatitis C virus RNA detection and HCV genotype in patients with chronic non-A, non-B hepatitis in Jakarta.

Antibody response in HCV infection may be variable and the variability of the serological response could be due to the differences in HCV strains. Since the distribution of hepatitis C virus genotype has been found to be geographically dependent, it is important to determine the distribution of HCV genotype in various countries with high prevalence of chronic non-A, non-B hepatitis. In this study, serum HCV RNA was examined in 53 patients suspected of chronic non-A, non-B hepatitis with an anti-HCV test as determined by currently available assay. HCV viremia was detected in 48 patients (90.6%). These patients had elevated serum ALT level at the time of HCV RNA determination. Using specific genotype probes, all isolates were classified into three different genotypes. Double and triple infections were also noted. HCV genotype 1b is the predominant genotype found in chronic hepatitis C patients in Jakarta.

Adult↗
PubMed1995

DNA fingerprinting of Musa cultivars with oligodeoxyribonucleotide probes specific for simple repeat motifs.

Using synthetic oligodeoxyribonucleotide probes against restriction-digested genomic DNA, we have established DNA fingerprinting of Musa cultivars. Of all the enzymes used, Eco RI and Hin dIII were found to be most informative, giving rise to individual specific band patterns with oligonucleotide probes of 15- to 18-base residues. Of the several probes and enzyme combinations used, the 15mer GACA probe with Eco RI and Hin dIII digests revealed a maximal level of polymorphism, and the probability of obtaining an identical band pattern between any two random genotypes was calculated to be 1.50 x 10(-9) and 1.59 x 10(-9), respectively. Oligonucleotide probes longer than 22 residues were also used but did not hybridize. The present approach is useful for cultivar identification and for overall genome analysis to establish relatedness among the various accessions of the Musa germplasm originating from different geographic locations. The relevance of using synthetic oligonucleotide probes based on simple repeat motifs for achieving DNA fingerprinting pattern is discussed.

Base Sequence↗