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S Patel

Publications and source records attributed to S Patel.

At least 289 records · Page 16Linked to original sources
PubMed1999

Antisense oligonucleotides to c-fos and c-jun inhibit intimal thickening in a rat vein graft model.

BACKGROUND: C-fos and c-jun are 2 immediate early genes that have been implicated in the stimulation of vascular smooth muscle cell proliferation and migration. In previous experiments in our laboratory with a rat vein graft model a 2- to 3-fold increase of messenger RNA of c-fos and c-jun were noted 1 hour after vein graft perfusion. Because c-fos and c-jun are up-regulated after the perfusion of vein grafts, the purpose of this study was to delineate the temporal expression of c-fos and c-jun protein and to study the effect of antisense oligonucleotides (ASO) to c-fos and c-jun on intimal thickening observed in this model. METHODS: Sprague-Dawley rats underwent bilateral interposition femoral artery grafts with use of the superficial epigastric vein, which was harvested from 15 minutes up to 2 weeks and analyzed by Western blot for Fos and Jun protein. Additional rats underwent bypasses and at the time of the procedure 1 graft was treated with a pluronic gel containing an ASO to c-fos, c-jun, or sense and the contralateral side was treated with pluronic gel only. The vein grafts were harvested 2 weeks after the procedure and perfusion fixed. After longitudinal sectioning, the intimal and total wall thicknesses were measured in the perianastamotic and midgraft regions by a morphometric digitizing microscope and the statistics were analyzed by a paired Student's t test. RESULTS: Protein analysis by Western blot showed that c-fos levels rose quickly within 2 hours and leveled at 6 hours 40-fold above basal levels after vein graft perfusion. Similarly, c-jun levels rose 10-fold above basal levels after 15 minutes and peaked at 2 hours 120-fold above basal levels. The treatment of the vein grafts with these ASOs resulted in a reduction of about 30% in the thickness of the intimal layer and the total wall thickness in both the perianastomotic and the midgraft regions, which was statistically significant different from control veins. CONCLUSION: These results indicate a possible therapeutic role for ASO to immediate early genes in the treatment of vein graft intimal hyperplasia.

Animals↗
PubMed1999

A fluorescent cholesterol analog traces cholesterol absorption in hamsters and is esterified in vivo and in vitro.

The fluorescent cholesterol analog 22-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3beta-ol (fluoresterol) was characterized as a tool for exploring the biochemistry and cell biology of intestinal cholesterol absorption. Hamsters absorbed fluoresterol in a concentration- and time-dependent manner, with an efficiency of about 15-30% that of cholesterol. Fluoresterol absorption was blocked by compounds known to inhibit cholesterol absorption, implying that fluoresterol interacts with those elements of the normal pathway for cholesterol absorption on which the inhibitors act. Confocal microscopy of small intestinal tissue demonstrated that fluoresterol was taken up by absorptive epithelial cells and packaged into lipoprotein particles, suggesting a normal route of intracellular trafficking. Uptake of fluoresterol was confirmed by biochemical analysis of intestinal tissue, and a comparison of [(3)H] cholesterol and fluoresterol content in the mucosa suggested that fluoresterol moved through the enterocytes more rapidly than did cholesterol. This interpretation was supported by measurements of fluoresterol esterification in the mucosa. Four hours after hamsters were given fluoresterol and [(3)H]cholesterol orally, 44% of the fluoresterol in the intestinal mucosa was esterified, compared to 8% of the [(3)H]cholesterol. Caco-2 cells took up 2- to 5-fold more [(3)H]cholesterol than fluoresterol from bile acid micelles, and esterified 21-24% of the fluoresterol but only 1-4% of the [(3)H]cholesterol. Thus fluoresterol apparently interacts with the proteins required for cholesterol uptake, trafficking, and processing in the small intestine.

Animals↗
PubMed1999

Enhancing the case manager's role through early identification of at-risk members.

This article describes a methodology to improve early identification and stratification of at-risk patients with comorbidities that could result in catastrophic illness. Data from the HMO line of business identified diabetic members. Total frequency and total cost for 19 comorbid diseases were calculated on data from the diabetic members who were classified as having high severity and comorbidity rankings. Members were further segmented into quadrants based on frequency and cost for referral to case management and/or educational programs. A total of 1312 HMO diabetic members were classified as high severity. Quadrant 4 diabetic members consumed greater than $10,000 in health care expenses and had 6 or more collapsed DECs. Similar types of comorbid diseases were found in both quadrants 2 and 4. Quadrant 2 diabetic members were identified as "in danger" members with the potential to incur total costs greater than $10,000.

Adult↗
PubMed1999

Temperature effects on surface pressure-induced changes in rat skin perfusion: implications in pressure ulcer development.

The effect of varying local skin temperature on surface pressure-induced changes in skin perfusion and deformation was determined in hairless fuzzy rats (13.5+/-3 mo, 474+/-25 g). Skin surface pressure was applied by a computer-controlled plunger with corresponding skin deformation measured by a linear variable differential transformer while a laser Doppler flowmeter measured skin perfusion. In Protocol I, skin surface perfusion was measured without heating (control, T=28 degrees C), with heating (T=36 degrees C), for control (probe just touching skin, 3.7 mmHg), and at two different skin surface pressures, 18 mmHg and 73 mmHg. Heating caused perfusion to increase at control and 18 mmHg pressure, but not at 73 mmHg. In Protocol II, skin perfusion was measured with and without heating as in Protocol I, but this time skin surface pressure was increased from 3.7 to 62 mmHg in increments of 3.7 mmHg. For unheated skin, perfusion increased as skin surface pressure increased from 3.7 to 18 mmHg. Further increases in surface pressure caused a decrease in perfusion until zero perfusion was reached for pressures over 55 mmHg. Heating increased skin perfusion for surface pressures from 3.7 to 18 mmHg, but not for pressures greater than 18 mmHg. After the release of surface pressure, the reactive hyperemia peak of perfusion increased with heating. In Protocol III, where skin deformation (creep and relaxation) was measured during the application of 3.7 and 18 mmHg, heating caused the tissue to be stiffer, allowing less deformation. It was found that for surface pressures below 18 mmHg, increasing skin temperature significantly increased skin perfusion and tissue stiffness. The clinical significance of these findings may have relevance in evaluating temperature and pressure effects on skin blood flow and deformation as well as the efficacy of using temperature as a therapeutic modality in the treatment of pressure ulcers.

Analysis of Variance↗
PubMed1999

The clinical anticalculus efficacy of a tartar control whitening dentifrice for the prevention of supragingival calculus in a three-month study.

The objective of this double-blind clinical study was to compare the effect of a new dentifrice (Colgate Tartar Control Plus Whitening Fluoride Toothpaste) for the prevention of supragingival calculus, with that of a commercially available calculus-inhibiting dentifrice (Crest Tartar Control Toothpaste). The study involved adult male and female subjects who had pre-qualified for participation by developing sufficient supragingival calculus (greater than 7.0 on the Volpe-Manhold Calculus Index) during an eight-week screening period. Subjects received a full oral prophylaxis, and were stratified into two treatment groups balanced for age, sex and qualifying calculus score. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for dental calculus were performed after twelve weeks' use of the study dentifrices, using the Volpe-Manhold Calculus Index, Fifty-eight (58) subjects complied with the protocol and completed the entire study. The Colgate Tartar Control Plus Whitening group exhibited a statistically significant (p < 0.001) 34.6% reduction in mean calculus score compared to the Crest Tartar Control group.

Adult↗
PubMed1998

Autocatalytic activation of human legumain at aspartic acid residues.

Human legumain was characterized following overexpression in a murine cell line as the C-terminal Ig-fusion protein. Upon acid treatment, the prolegumain autoproteolyzed distal to two aspartic acid residues to yield a highly active form. The ability of mature legumain to cleave after aspartic acid residues was confirmed with a small peptide substrate. Substitution of alanine for the putative catalytic cysteine, or for either of two strictly conserved histidine residues, partly or wholly eliminated autoactivation but not the ability of wild-type legumain to correctly process the variants to the properly sized proteins.

Amino Acid Sequence↗
PubMed1998

Phytochromes and photomorphogenesis in Arabidopsis.

Plants have evolved exquisite sensory systems for monitoring their light environment. The intensity, quality, direction and duration of light are continuously monitored by the plant and the information gained is used to modulate all aspects of plant development. Several classes of distinct photoreceptors, sensitive to different regions of the light spectrum, mediate the developmental responses of plants to light signals. The red-far-red light-absorbing, reversibly photochromic phytochromes are perhaps the best characterized of these. Higher plants possess a family of phytochromes, the apoproteins of which are encoded by a small, divergent gene family. Arabidopsis has five apophytochrome-encoding genes, PHYA-PHYE. Different phytochromes have discrete biochemical and physiological properties, are differentially expressed and are involved in the perception of different light signals. Photoreceptor and signal transduction mutants of Arabidopsis are proving to be valuable tools in the molecular dissection of photomorphogenesis. Mutants deficient in four of the five phytochromes have now been isolated. Their analysis indicates considerable overlap in the physiological functions of different phytochromes. In addition, mutants defining components acting downstream of the phytochromes have provided evidence that different members of the family use different signalling pathways.

Arabidopsis↗
PubMed1998

Correlates of residential wiring code used in studies of health effects of residential electromagnetic fields.

The home wiring code is the most widely used metric for studies of residential electromagnetic field (EMF) exposure and health effects. Despite the fact that wiring code often shows stronger correlations with disease outcome than more direct EMF home assessments, little is known about potential confounders of the wiring code association. In a study carried out in southern Connecticut in 1988-1991, the authors used strict and widely used criteria to assess the wiring codes of 3,259 homes in which respondents lived. They also collected other home characteristics from the tax assessor's office, estimated traffic density around the home from state data, and interviewed each subject (2,967 mothers of reproductive age) for personal characteristics. Women who lived in very high current configuration wiring coded homes were more likely to be in manual jobs and their homes were older (built before 1949, odds ratio (OR) = 73.24, 95% confidence interval (CI) 29.53-181.65) and had lower assessed value and higher traffic densities (highest density quartile, OR = 3.99, 95% CI 1.17-13.62). Because some of these variables have themselves been associated with health outcomes, the possibility of confounding of the wiring code associations must be rigorously evaluated in future EMF research.

Adult↗
PubMed1998

2-Nitrophenylcarbamoyl-(S)-prolyl-(S)-3-(2-naphthyl)alanyl-N-benzyl-N - methylamide (SDZ NKT 343), a potent human NK1 tachykinin receptor antagonist with good oral analgesic activity in chronic pain models.

A lead compound which had sub-micromolar affinity for the rabbit NK1 receptor but negligible affinity for rat NK1 receptors, 3a, was discovered by directed screening. 2-Substitution in the ring of the benzylthiourea substituent in the initial lead was found to be important, and halogens (Cl, Br) in this position were found to improve affinity for the human receptor. The activity of a series of 2-halo-substituted benzylthioureas was then optimized by modification of the proline diphenylmethyl amide, guided by a simple conceptual model based on structural overlay between these early antagonists and NK1 selective peptides. In this way, aromatic amino acid amides were identified which had improved affinity with respect to the starting diphenylmethyl (DPM) amides. The first sub-nanomolar ligand for the human NK1 receptor which arose from this series, 4af, combined a 2-chlorobenzylthiourea unit with a 2-naphthylalanine amide. Contemporaneously it was discovered that the benzylthiourea unit could be simplified to a phenylthiourea providing that an appropriate 2-substituent was also incorporated. Combination of these two series gave 2-NO2 phenylthiourea analogues which led directly to the analogous urea, 5f (2-nitrophenylcarbamoyl-(S)-prolyl-(S)-3-(2-naphthyl)alanyl-N-benz yl- N-methylamide, SDZ NKT 343), a highly potent ligand for the human NK1 receptor (Ki = 0.16 nM). In addition to its high in vitro potency, 5f proved to be a potent orally active analgesic in guinea pig models of chronic inflammatory and neuropathic pain. The nature of the 2-aryl substituent was found to be critical for oral activity in this series. Clinical evaluation of 5f as a novel analgesic agent is currently underway.

Analgesics↗
PubMed1998

A regulatory role of sulfhydryl groups in modulation of sperm membrane conformation by heavy metals: sulfhydryl groups as markers for infertility assessment.

Sperm membrane sulfhydryl groups, when masked by heavy metals like cobalt and copper at very low concentration (10(-9)) shows inhibition of lipid peroxidation and superoxide dismutase activity. Inhibition of lipid peroxidation is suggested to lead to reduction in membrane fluidity, a prerequisite for normal sperm function. Augmentation of lipid peroxidation by pentoxifylline in oligospermia provides evidence to this hypothesis that membrane sulfhydryls play a regulatory role in membrane modulation. It is therefore suggested that these sulfhydryl groups can be used as a tool for infertility assessment in unexplained male infertility.

Animals↗
PubMed1998

Fibroblast growth factor-2 (FGF-2) induces vascular endothelial growth factor (VEGF) expression in the endothelial cells of forming capillaries: an autocrine mechanism contributing to angiogenesis.

FGF-2 and VEGF are potent angiogenesis inducers in vivo and in vitro. Here we show that FGF-2 induces VEGF expression in vascular endothelial cells through autocrine and paracrine mechanisms. Addition of recombinant FGF-2 to cultured endothelial cells or upregulation of endogenous FGF-2 results in increased VEGF expression. Neutralizing monoclonal antibody to VEGF inhibits FGF-2-induced endothelial cell proliferation. Endogenous 18-kD FGF-2 production upregulates VEGF expression through extracellular interaction with cell membrane receptors; high-Mr FGF-2 (22-24-kD) acts via intracellular mechanism(s). During angiogenesis induced by FGF-2 in the mouse cornea, the endothelial cells of forming capillaries express VEGF mRNA and protein. Systemic administration of neutralizing VEGF antibody dramatically reduces FGF-2-induced angiogenesis. Because occasional fibroblasts or other cell types present in the corneal stroma show no significant expression of VEGF mRNA, these findings demonstrate that endothelial cell-derived VEGF is an important autocrine mediator of FGF-2-induced angiogenesis. Thus, angiogenesis in vivo can be modulated by a novel mechanism that involves the autocrine action of vascular endothelial cell-derived FGF-2 and VEGF.

3T3 Cells↗
PubMed1998

Mutational analysis of structural features of rat hormone-sensitive lipase.

Hormone-sensitive lipase (HSL) is a cytosolic neutral lipase that hydrolyzes intracellular stores of triacylglycerols and cholesteryl esters. HSL activity is regulated via phosphorylation-dephosphorylation, with cyclic AMP-dependent protein kinase increasing activity following phosphorylation of a single serine and Ca2+/calmodulin-dependent protein kinase II phosphorylating another serine at a basal site. The current studies used site-directed mutagenesis to show that Ser-563 of rat HSL is phosphorylated by cyclic AMP-dependent protein kinase and that Ser-565 is phosphorylated by Ca2+/calmodulin-dependent protein kinase II. Mutation of Ser-563-->Ala eliminated HSL hydrolytic activity against cholesteryl ester, triacylglycerol, and diacylglycerol substrates to the same extent as mutation of Ser-423-->Ala, the presumed catalytic site. Mutation of Ser-565-->Ala modestly decreased HSL activity. In contrast, mutation of Ser-563-->Asp preserved HSL hydrolytic activity and even increased activity 20% above the control wild-type enzyme. Molecular modeling of the catalytic pocket of HSL suggested the involvement of Val-710. Mutation of Val-710-->Ala resulted in an 85% loss of HSL hydrolytic activity. The results of these studies illustrate the importance of the presence of a hydroxyl group or negative charge at residue 563, either for proper conformation of rat HSL or for proper stabilization of substrate to allow maintenance of hydrolytic activity, as well as the importance of the involvement of additional amino acids in the catalytic pocket of the enzyme.

Alanine↗
PubMed1998

Dangers of delay of initiation of either thrombolysis or primary angioplasty in acute myocardial infarction with increasing use of primary angioplasty.

We observed treatment delays and suboptimal outcomes when beginning to treat patients with acute myocardial infarction (AMI) with primary angioplasty. Of the 37 patients treated during a 12-month period, 12 (32%) required either emergency bypass surgery or died. Delayed time intervals to balloon reperfusion (mean 134 minutes) probably contributed to these adverse outcomes, with hemodynamic instability requiring pressors or intra-aortic balloon pumping in 15 patients, 12 (75%) before the first balloon inflation. Eleven of the 12 patients with significant adverse outcomes required such intervention. As angioplasty use increased, time intervals to thrombolysis in those not treated with angioplasty increased from an average of 29 minutes (53% treated less than the national standard of 30 minutes) to 39 minutes (32% treated <30 minutes, p <0.001). During the last 2 months of the study period, the time intervals had increased to 48 minutes (14% treated <30 minutes, p <0.0001). There was no change in thrombolytic time intervals at a local community hospital that did not offer primary angioplasty. Emergency Department physician confusion about the best therapy (angioplasty or thrombolysis) was documented in the medical records in 42% of cases (53 of 127). Confusion regarding therapy of AMI led to unacceptable delays in the administration of thrombolytic agents and probably contributed to the adverse outcomes in patients receiving primary angioplasty.

Aged↗
PubMed1998

Novel natural product 5,5-trans-lactone inhibitors of human alpha-thrombin: mechanism of action and structural studies.

High-throughput screening of methanolic extracts from the leaves of the plant Lantana camara identified potent inhibitors of human alpha-thrombin, which were shown to be 5,5-trans-fused cyclic lactone euphane triterpenes [O'Neill et al. (1998) J. Nat. Prod. (submitted for publication)]. Proflavin displacement studies showed the inhibitors to bind at the active site of alpha-thrombin and alpha-chymotrypsin. Kinetic analysis of alpha-thrombin showed tight-binding reversible competitive inhibition by both compounds, named GR133487 and GR133686, with respective kon values at pH 8.4 of 1.7 x 10(6) s-1 M-1 and 4.6 x 10(6) s-1 M-1. Electrospray ionization mass spectrometry of thrombin/inhibitor complexes showed the tight-bound species to be covalently attached, suggesting acyl-enzyme formation by reaction of the active-site Ser195 with the trans-lactone carbonyl. X-ray crystal structures of alpha-thrombin/GR133686 (3.0 A resolution) and alpha-thrombin/GR133487 (2.2 A resolution) complexes showed continuous electron density between Ser195 and the ring-opened lactone carbonyl, demonstrating acyl-enzyme formation. Turnover of inhibitor by alpha-thrombin was negligible and mass spectrometry of isolated complexes showed that reversal of inhibition occurs by reformation of the trans-lactone from the acyl-enzyme. The catalytic triad appears undisrupted and the inhibitor carbonyl occupies the oxyanion hole, suggesting the observed lack of turnover is due to exclusion of water for deacylation. The acyl-enzyme inhibitor hydroxyl is properly positioned for nucleophilic attack on the ester carbonyl and therefore relactonization; furthermore, the higher resolution structure of alpha-thrombin/GR133487 shows this hydroxyl to be effectively superimposable with the recently proposed deacylating water for peptide substrate hydrolysis [Wilmouth, R. C., et al. (1997) Nat. Struct.Biol. 4, 456-462], suggesting the alpha-thrombin/GR133487 complex may be a good model for this reaction.

Acylation↗
PubMed1998

T cell epitopes of insulin defined in HLA-DR4 transgenic mice are derived from preproinsulin and proinsulin.

Approximately one-half of Caucasians with newly diagnosed insulin-dependent diabetes mellitus (IDDM) have autoantibodies to insulin, and the majority of those express the HLA-DR4 genotype [Ziegler, R., Alper, C. A., Awdeh, Z. L., Castano, L., Brink, S. J., Soeldner, J. S., Jackson, R. A. & Eisenbarth, G. S. (1991) Diabetes 40, 709-714]. However, it has been difficult to demonstrate T cell proliferative responses to human insulin in IDDM patients [Durinovic-Bello, I., Hummel, M. & Ziegler, A. G. (1996) Diabetes 45, 795-800]. We have immunized transgenic mice expressing the susceptible HLA-DR (alpha1*0101,beta1*0401) (hereafter called DRB1*0401) and human CD4 molecules on a murine major histocompatibility complex class II null background, with human preproinsulin (PPI), proinsulin (PI), and insulin and derived large panels of T cell hybridomas to determine the immunogenic epitopes of these proteins. These results show that the prohormones PI or PPI carry the major immunogenic T cell epitope in the DRB1*0401 transgenic mice. The PPI/PI immunodominant epitope LALEGSLQK was localized at the C-peptide/A-chain junction. This T cell epitope PPI/PI LALEGSLQK is unusual because, normally, it is proteolytically destroyed during the maturation of the insulin molecule. Additionally, this T cell epitope is both processed and presented by human DRB1*0401-positive Epstein-Barr virus transformed B cells, and it can also stimulate T cells from the peripheral blood of HLA-DR4-positive patients with type 1 diabetes. These findings may partly explain why susceptibility to type 1 diabetes is associated with HLA-DR4-positive individuals and why T cell responses to the mature insulin protein are rarely detected in IDDM patients.

Amino Acid Sequence↗