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Biomedical subjects

R Shankar

Publications and source records attributed to R Shankar.

At least 19 recordsLinked to original sources
PubMed1995

Bi-directional stimulation of adherence to extracellular matrix components by human head and neck squamous carcinoma cells and endothelial cells.

Head and neck squamous cell carcinoma (HNSCC) cultures were established from two patients to determine if cancer and endothelial cells bi-directionally regulate their adherence to extracellular matrix components (ECM), an important process for tumor vascularization and metastasis. Soluble products from endothelial cells transiently enhanced adherence by HNSCC to ECM and increased surface levels of beta 1 and beta 4 integrins, although not beta 3. HNSCC products enhanced endothelial cell adherence to fibronectin and laminin, and beta 1 and beta 4 expression. These data show bi-directional enhancement of adherence to ECM and integrin expression among endothelial and tumor cells, which may facilitate metastasis and neovascularization.

Animals↗
PubMed1995

Gender differences in disability: a comparison of married patients with schizophrenia.

Gender differences in disability constitute a fertile area of research, as disabilities need to be measured and evaluated in the social context which defines role expectations and consequently the role performance. This paper reports on the differences in disability in married patients with schizophrenia, as marital status is an important determinant of role expectation. The study sample constituted 30 married patients, of both sexes, who satisfied DSM-III criteria for schizophrenia, and were living with their spouse at the time of assessment. Disability was evaluated using the DAS (modified version). The findings indicated that women were more disabled than men on many of the evaluation parameters (p < 0.05); there was also a strong correlation between negative symptoms and disability variables in both the sexes. While a correlation between PSE syndromes and disability variables was seen in the case of males, the relationship was not seen in females. Stepwise regression also revealed that negative symptoms predominated among the factors associated with global disability in both sexes. Most of the reports in the literature reveal that women are less disabled than men. The findings of this study, that women are more disabled than men, is discussed in the context of the social conditions prevailing in India.

Adult↗
PubMed1995

TNF-alpha induces peroxynitrite-mediated depletion of lung endothelial glutathione via protein kinase C.

We tested the hypothesis that tumor necrosis factor-alpha (TNF) induces a peroxynitrite (ONOO-)-mediated depletion of glutathione via a protein kinase C (PKC)-dependent mechanism in pulmonary artery endothelial monolayers (PAEM). PAEM were incubated with TNF (1,000 U/ml) for 6 and 18 h. The PAEM were assayed for ONOO(-)-dependent changes in the concentration of luminol, free glutathione [Gfree; i.e., reduced glutathione and oxidized glutathione (GSSG)] and GSSG. TNF treatment decreased luminol and Gfree, and increased GSSG and GSSG/Gfree, compared with treatment with control media. The TNF-induced effects were prevented by co-incubation with the nitric oxide synthase inhibitors NG-monomethyl-L-arginine (1 mM), NG-nitro-L-arginine methyl ester (1 mM), or NG-nitro-L-arginine (1 mM). In addition, the TNF-induced effects were prevented by superoxide dismutase (10 U/ml), which removes O2-, and by urate (0.5 mM) and L-cysteine (3 mM), putative scavengers of ONOO-. The treatment of PAEM with the PKC activator phorbol 12-myristate 13-acetate (PMA, 1 microM) induced similar alterations in luminol and glutathione as TNF. TNF and PMA induced a protein of similar molecular weight (approximately 90 kDa) in the focal contact-rich fraction of PAEM lysate. TNF- and PMA-induced effects were prevented with the specific PKC inhibitor calphostin C (1 microM). The data indicate that TNF-induced PKC activation mediates ONOO- generation, which results in the oxidation and depletion of glutathione in PAEM.

Animals↗
PubMed1995

Down regulation of CD11b and CD18 expression in atherosclerotic lesion-derived macrophages.

Monocyte/macrophages play an important role in the development of atherosclerosis. Electron microscopic evidence suggests that in the early stages, lipid laden monocytes leave the lesion to reenter the circulation. This reverse monocyte traffic ceases as the lesion develops. We hypothesize that monocyte/macrophages may not be able to exit the lesion and reenter the circulation because of the reduced expression of CD11/CD18 integrins. We have compared CD11b and CD18 expression of peripheral blood monocytes from normal rabbits (NMø) to atherosclerotic lesion-derived macrophages (AthMø) by anti-CD11b and anti-CD 18 antibody staining, followed by flow cytometry and immunohistochemical staining. AthMø were isolated from aortic lesions of rabbits fed 2 per cent cholesterol diet following balloon angioplasty. AthMø were separated into two regions based on their size and granularity by flow cytometry. All macrophages stained positively with RAM 11. Our results indicated that NMø showed a strong cell surface expression of CD11b and CD18. The less granular and smaller AthMø showed little anti-CD11b or anti-CD18 antibody staining, indicating very little CD11b or CD18 antibody staining, indicating very little CD11b or CD18 expression. The more granular and larger cells showed surface expression of both CD11b and CD18. With respect to CD18, over 90 per cent of NMø expressed CD18, only 37 per cent of the large granular AthMø and less than 1 per cent of the smaller, less granular AthMø stained positive for CD18. Immunohistochemical studies revealed strong surface expression of CD11b and CD18 on normal monocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Angioplasty, Balloon↗
PubMed1994

Thrombin receptor-activating peptides differentially stimulate platelet-derived growth factor production, monocytic cell adhesion, and E-selectin expression in human umbilical vein endothelial cells.

Recent studies have shown that the synthetic peptides SFL LRN and SFL LRN PND KYEPF (thrombin receptor-activating peptides (TRAP)) derived from the deduced sequence of the new amino terminus of the cleaved thrombin receptor can mimic thrombin receptor activation, act as full agonists for platelet activation, and induce prostaglandin I2 production as well as cytosolic Ca2+ increase in human umbilical vein endothelial cells (HUVEC). Here, we have compared the ability of these synthetic peptide ligands and thrombin to stimulate platelet-derived growth factor (PDGF) production by, and monocyte adhesion to, HUVEC. Thrombin (50 units/ml) and TRAP (25 microM) maximally stimulated monocyte adhesion. Furthermore, the stimulation of E-selectin cell surface expression and the steady-state E-selectin mRNA levels by thrombin and TRAP were comparable. Thrombin (50 units/ml) stimulated PDGF production 400% above the basal level in 24 h, whereas the 6-mer and 14-mer TRAP, even at 200 microM, did not significantly stimulate PDGF production. Northern analysis, however, revealed that TRAP at 100 microM stimulated PDGF-A and -B chain mRNA expression to a level similar to that induced by thrombin. These results suggest that activation of cell signaling by TRAP can mimic thrombin and is sufficient for the stimulation of monocyte adhesion to HUVEC; however, thrombin-stimulated PDGF production by HUVEC may require mechanisms in addition to the signaling events initiated by TRAP or may require the participation of a novel thrombin receptor.

Amino Acid Sequence↗
PubMed1992

3-Deazaadenosine inhibits thrombin-stimulated platelet-derived growth factor production and endothelial-leukocyte adhesion molecule-1-mediated monocytic cell adhesion in human aortic endothelial cells.

Injury to the vascular endothelium and the subsequent inflammatory response are considered prerequisites for the development of atherosclerosis. Platelet-derived growth factor (PDGF) production by and monocyte adhesion to aortic endothelial cells (EC) may participate in this inflammatory process and therefore are two potential targets for control by anti-inflammatory agents. Our previous studies have demonstrated that monocyte adhesion and PDGF production are stimulated by thrombin in EC. Here, we provide evidence that treatment of EC with the anti-inflammatory agent 3-deazaadenosine (c3Ado) effectively abolished thrombin-stimulated PDGF production and monocyte adhesion. c3Ado had no significant effect on either basal monocyte adhesion or constitutive PDGF production. c3Ado was also effective in negating monocyte adhesion induced by other agonists, such as interleukin-1, phorbol 12-myristate 13-acetate (PMA), and lipopolysaccharide. Northern analysis demonstrated that c3Ado significantly reduced thrombin- and PMA-stimulated steady-state levels of PDGF-A chain, PDGF-B chain, and endothelial-leukocyte adhesion molecule-1 (ELAM-1) mRNAs. Nuclear run-on studies demonstrated that a marked transcriptional activation of these genes by thrombin and PMA was abrogated by c3Ado treatment. The transcriptional rate of the alpha-tubulin gene was unaffected by the drug. Antibody binding studies with an anti-ELAM-1 monoclonal antibody 7A9 revealed that thrombin-stimulated EC expression of ELAM-1 was abolished by c3Ado, indicating that the suppression of ELAM-1 expression on EC surface may be a mechanism by which c3Ado interferes with monocyte adhesion. Experiments with the nucleoside transport inhibitor nitrobenzylthioinosine suggested that the transport of c3Ado into EC was required for its inhibitory activity. In addition, L-homocysteine thiolactone was found to potentiate the inhibitory activity of c3Ado, suggesting that the accumulation of intracellular c3Ado homocysteine may be the underlying mechanism by which c3Ado inhibits thrombin-induced EC function. Taken together, these results indicate that c3Ado may prove effective against vascular injury and inflammation through its ability to inhibit induction of both monocyte adhesion and PDGF production.

Adenine Nucleotides↗
PubMed1992

Thrombin stimulates PDGF production and monocyte adhesion through distinct intracellular pathways in human endothelial cells.

Thrombin stimulates multiple functions in cultured endothelial cells (EC), including an increase in cell surface adhesion sites for monocytes and the production of platelet-derived growth factor (PDGF). We have initiated studies to define the intracellular signaling pathways involved in these two thrombin-induced EC functions by focusing on the possible roles of the Na(+)-H+ antiporter and guanine nucleotide-binding proteins (G proteins). Amiloride suppressed thrombin-stimulated PDGF production by human aortic EC without affecting either basal PDGF production or overall protein synthesis. The steady-state mRNA levels of PDGF-A and PDGF-B chain were not reduced by amiloride. In replicate EC cultures, amiloride had no effect on thrombin-stimulated monocyte adhesion. In addition, thrombin induction of PDGF production, but not monocyte adhesion, was abrogated in the absence of extracellular sodium. Thrombin stimulation of both monocyte adhesion and PDGF production appeared to involve a pertussis toxin-insensitive G protein. Thrombin induced an increase in [35S]guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) binding to human EC membranes. GTP gamma S, in the presence of a suboptimal concentration of thrombin, caused maximal stimulation of both monocyte adhesion and PDGF production. The effect of GTP gamma S on PDGF production was at the level of transcription. These results indicate that the EC is capable of responding to a pluripotent agonist such as thrombin through multiple signaling pathways, which converge and diverge to achieve differential cellular responses.

Amiloride↗
PubMed1991

Expression of human aldose and aldehyde reductases. Site-directed mutagenesis of a critical lysine 262.

Human aldose reductase (EC 1.1.1.21) and aldehyde reductase (EC 1.1.1.2) are implicated in the development of diabetic complications by a variety of mechanisms, and a number of drugs to inhibit these enzymes have been proposed for the therapy and prevention of these complications. To probe the structure and function of these two enzymes, we used site-directed mutagenesis in the cDNAs of both enzymes to replace lysine 262 with methionine. Wild-type and mutant enzymes were overexpressed in Escherichia coli and purified by anion exchange and affinity chromatography. N-terminal sequence analysis, Western blots, and kinetic studies confirmed the identity of the recombinant wild-type enzymes with the native human placental and liver enzymes. Recombinant aldose reductase (hAR) and aldehyde reductase (hGR) have apparent kinetic constants virtually identical to their respective native enzymes. The mutant aldose reductase (hARK262 greater than M) shows a 66-fold increase in Km for NADPH with respect to the wild type (1.9 +/- 0.4 microM versus 125 +/- 14 microM), whereas the Km for DL-glyceraldehyde increased 35-fold (20 +/- 2 versus 693 +/- 41 microM). The same constants for the mutant aldehyde reductase (hGRK262 greater than M) increased 97- and 86-fold, respectively (from 2.0 +/- 0.4 to 194 +/- 16 microM and from 1.6 +/- 0.4 to 137 +/- 3 mM). These results indicate that lysine 262 in aldose reductase and aldehyde reductase is crucial to their catalytic activity by affecting co-factor binding.

Aldehyde Reductase↗
PubMed1991

Noninvasive measurement of compliance of human leg arteries.

Electrical impedance plethysmography has been evaluated for early detection of peripheral atherosclerosis. A pressure cuff was wrapped around the lower leg and the cuff pressure increased. Two circumferential electrodes glued in the middle of the cuff recorded the impedance pulse, from which the arterial pulse volume was calculated. The ratio of maximal arterial volume change to the pulse pressure was determined as a measure of maximal compliance Cp. Based on the data from 118 human subjects, Cp was found to correlate well with known cardiovascular risk factors. For example, Cp decreased on the average from 3.08 to 1.92 microL.mm Hg-1.cm-1 (1 microL.mm Hg-1.cm-1 = 7.5 x 10(-10) m4.N-1) in groups of subjects of increasing age from 22 to 70 years. Subjects on a regular exercise program had an average value of 3.86, while those with proven peripheral vascular disease had a value of 0.70. In a related pathologic validation study on 15 monkeys fed a cholesterol-control diet a good correlation was found between the limb peak compliance and morphometric data obtained from iliac and carotid arteries.

Adult↗
PubMed1990

Correlation of noninvasive arterial compliance with anatomic pathology of atherosclerotic nonhuman primates.

Fifteen cynomolgus monkeys were fed a control (n = 3) or cholesterol-containing diet (n = 12) for 26 months. An impedance plethysmograph and a calibrated volume plethysmograph were used to determine femoral arterial volume change, delta V, from femoral arteries. Abdominal aorta pulse pressure was measured directly. The ratio of delta V to pulse pressure, delta P, defined femoral artery compliance. At necropsy, sections of pressure fixed common iliac, external iliac, femoral and carotid arteries were obtained for measurements. The site of the largest atherosclerotic lesion was chosen for histologic measurement. The percentage intima in arterial tissue (PIAT) was determined at the site of the largest atherosclerotic lesions in iliac arteries and averaged. Peak compliance was inversely correlated with the PIAT (Pearson correlation -0.39, P less than 0.10, n = 21) which improved to r = -0.52, P less than 0.01 (n = 27) when the PIAT also included the carotid atherosclerosis. The correlation coefficients were comparable to those obtained for total plasma cholesterol concentration, and direct systolic and diastolic blood pressure. Peak compliance and alpha-cholesterol concentrations provided the best multiple linear regression fit for the prediction of PIAT.

Animals↗
PubMed1990

Model reference adaptive control with constraints for postoperative blood pressure management.

This paper presents an approach for adaptive control of blood pressure using sodium nitroprusside. A modified stochastic model reference adaptive control algorithm with a time varying reference model is developed and used for this purpose. An automatic adjustment of the reference model is proposed in order to optimize the performance of the closed loop system while meeting clinical constraints imposed on the infusion rate and the mean arterial pressure. Extensive computer simulations show the robustness of the proposed controller in the presence of relatively high noise levels, over the full range of plant parameters and for large parameter variations.

Algorithms↗
PubMed1989

Influence of probucol on early experimental atherogenesis in hypercholesterolemic rats.

A possible cellular action for probucol in early atherogenesis was investigated. In diet-induced hypercholesterolemic rats, probucol reduced aortic accumulation of cholesterol without ameliorating monocyte attachment to the arterial endothelium. Under the imposed conditions, circulating cholesterol levels were not significantly altered by probucol. 125I-labeled acetyl LDL uptake and degradation studies with mouse peritoneal macrophages revealed that probucol had an inhibitory effect on the scavenger receptor pathway. The data suggested that the observed beneficial effects of probucol were not related to an early cholesterol-induced injury phase which might involve calcium. Instead, probucol probably through its free radical scavenging property, intervened at a subsequent cellular level to restrict lipid accumulation.

Animals↗
PubMed1989

Protection of crystal-induced polymorphonuclear leukocyte membranolysis by phosphocitrate.

The protection afforded by phosphocitrate, a phosphorylated polycarboxylic acid, against crystal-induced membrane damage to polymorphonuclear leukocytes was studied in vitro. Membranolysis was assessed by nitro blue tetrazolium salt reduction, lactate dehydrogenase release, and scanning electron microscopy. Phosphocitrate protected strongly against hydroxyapatite crystal-induced damage, an action attributable to crystal surface binding of phosphocitrate rather than to the membrane. The ability of phosphocitrate to prevent hydroxyapatite crystallization, together with its membrane protective effect against preformed crystals, would suggest that the compound might have a useful future role against crystal-induced arthropathies.

Apatites↗
PubMed1988

Retardation of calcification of bovine pericardium used in bioprosthetic heart valves by phosphocitrate and a synthetic analogue.

The purpose of this study was to determine if phosphocitrate (PC), a naturally occurring inhibitor of calcification, and its synthetic analogue, N-sulpho-2-amino tricarballylate (SAT), administered either by daily injection or local delivery via Alzet osmotic minipump, could inhibit calcification of glutaraldehyde-preserved bovine pericardium used in bioprosthetic heart valves, subcutaneously implanted in rats. Local drug delivery, but not systemic administration, was effective. PC, administered by Alzet minipump (12 mg.kg-1.day-1), inhibited calcification significantly (tissue calcium = 5 +/- 2 micrograms/mg dry tissue, mean +/- SEM), compared with untreated or saline-treated controls (89 +/- 9 and 49 +/- 9 micrograms/mg, respectively). SAT, administered by the same route at both the same and a higher molar dosage, was less potent (tissue calcium = 26 +/- 9 micrograms/mg and 17 +/- 5 micrograms/mg, respectively). PC and SAT therapy were not associated with adverse effects. We conclude that locally administered PC and SAT can inhibit intrinsic calcification of bovine pericardium, with PC being more potent.

Animals↗