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M Patel

Publications and source records attributed to M Patel.

At least 325 records · Page 18Linked to original sources
PubMed1993

Clarification of the pH-dependent kinetic behaviour of papain by using reactivity probes and analysis of alkylation and catalysed acylation reactions in terms of multihydronic state models: implications for electrostatics calculations and interpretation of the consequences of site-specific mutations such as Asp-158-Asn and Asp-158-Glu.

1. The complex behaviour of papain (EC 3.4.22.2) in acidic media has been investigated by (a) stopped-flow reactivity probe kinetics using 4,4'-dipyrimidyl disulphide (I) and 2,2'-dipyridyl disulphide (II) as thiol-specific time-dependent inhibitors with markedly different susceptibilities to activation by hydronation (protonation) and (b) using the multitasking application program SKETCHER for the rapid evaluation of pH-dependent kinetic data by means of interactive manipulation of calculated curves. 2. The substantially lower basicity of (I) (pKa 0.91) than that of (II) (pKa 2.45) combined with retention of high reactivity permitted the pKa for the formation of the (Cys-25)-S-/(His-159)-Im+H ion-pair state of papain to be determined kinetically as 3.4, a value close to that (3.3) deduced by potentiometric difference titration [Lewis, Johnson and Shafer (1976) Biochemistry 15, 5009-5017] and lower than the value (approx. 4) often reported from pH-dependent kinetic studies. The higher values are now known to arise from inadequate data analysis that does not take account of other overlapping kinetically influential ionizations. 3. Re-evaluation of the extensive sets of pH-kcat/Km data for the hydrolysis of nine substrates by papain reported by Polgár and Halász (1978) (Eur. J. Biochem. 88, 513-521) by making use of SKETCHER, the known pKa value (3.4) from the reaction with compound (I) and two additional kinetically influential pKa values deduced from the reaction with compound (II) now permits the identification of the pH-dependent events in reactions of papain with inhibitors and substrates. 4. A major conclusion is that, whereas in reactions of simple alkylating agents and compound (I) full nucleophilic character of (Cys-25)-S-/(His-159)-Im+H is provided by hydronic dissociation with pKa 3.3-3.4, in catalysis relatively little catalytic competence is produced consequent upon ion-pair formation. Substantial catalytic competence requires further hydronic dissociation with pKa approx. 4, and for cationic substrates further enhancement is produced by hydronic dissociation with pKa approx. 5. 5. The present work, together with the kinetic analysis of reactions of papain in alkaline media reported by Mellor, Thomas, Topham and Brocklehurst [Biochem. J. (1993) 290, 289-296], defines the kinetically influential ionizations of papain as 3.4, 4.0, 5.0, 8.3 and 10.0 of which 3.4 and 8.3 relate to the formation and subsequent dehydronation of the ion-pair state.(ABSTRACT TRUNCATED AT 400 WORDS)

Acylation↗
PubMed1993

Abnormal growth regulation of vascular smooth muscle cells by heparin in patients with restenosis.

Proliferation of vascular smooth muscle cells (VSMC) underlies myointimal hyperplasia, which can lead to restenosis after angioplasty and vascular surgery. We propose that some individuals have an intrinsic capacity for this exaggerated response to vascular injury, partly through decreased sensitivity to the physiological growth inhibitor heparin. We investigated the effect of heparin on VSMC from restenotic lesions and from apparently normal vessels of the same patients, and VSMC from control patients undergoing primary bypass procedures. Cells from patients with restenosis (both restenotic lesion and undiseased vein) showed much lower sensitivity to growth inhibition by heparin than the controls (median inhibition 8 [95% Cl -2 to 25] vs 22 [15-44]%, p < 0.001); this finding suggests aberrant growth regulation in these cells.

Arterial Occlusive Diseases↗
PubMed1993

Unifocal Langerhans' cell histiocytosis (eosinophilic granuloma) of the petrous apex.

Eosinophilic granuloma is a localized form of histiocytosis X, or Langerhans' cell histiocytosis, a benign lesion of proliferating Langerhans' cells. It is the least severe of the histiocytosis syndromes, and is characterized by lytic lesions of one or more bones. Temporal bone lesions usually occur in association with multifocal disease; however, isolated lesions may occur in either the mastoid bone alone, or in the entire temporal bone, without disease elsewhere in the body. We present the first case (to our knowledge) of eosinophilic granuloma, or unifocal Langerhans' cell histiocytosis, limited to the petrous apex. The patient, an 8-year-old girl, presented with facial nerve paralysis. Because of delay in definitive diagnosis, the disease extended beyond its original boundaries and resulted in complete destruction of the temporal bone. We will also review Langerhans' cell histiocytosis, with attention to its involvement of the temporal bone.

Child↗
PubMed1993

Influence of debrisoquine hydroxylation phenotype on the pharmacokinetics of mexiletine.

Marked interindividual variation has been observed in the pharmacokinetics of the antiarrhythmic agent mexiletine. The fact that its urinary excretion is dependent on urinary pH may account, in part, for such variation. The influence that genetic differences in hepatic metabolism of the debrisoquine-type may have on mexiletine pharmacokinetics was considered in this study. The pharmacokinetics and urinary excretion of mexiletine (250 mg administered intravenously) were investigated in 5 rapid extensive metabolisers (EM), 5 slow EM and 5 poor metabolisers (PM) of debrisoquine, under conditions of controlled urinary pH. Mexiletine disposition kinetics was found to be altered in PM individuals. These subjects showed higher total area under the curve (AUC), (15.7 versus 8.16 micrograms.h.ml-1) prolonged elimination half-lives (in serum and urine) (serum: 18.5 versus 11.6 h, urine: 19.2 versus 11.7 h) and lower total clearance values compared with EM (216 versus 450 ml.min-1). In this respect, slow EM individuals generally presented intermediate values of those pharmacokinetic parameters. A higher incidence of adverse-effects was also observed among slow EM and PM subjects. It is concluded that genetic differences in mexiletine oxidation of the debrisoquine-type have an influence on its observed pharmacokinetic variability. The clinical consequences are discussed.

Adult↗
PubMed1993

Posture and the spread of hyperbaric bupivacaine in parturients using the combined spinal epidural technique.

This study was undertaken to analyze the effect of posture on the spread of hyperbaric bupivacaine in pregnant women using a combined spinal extradural technique, and to assess the quality of analgesia provided by 10 mg bupivacaine when using this technique. Fifty parturients undergoing elective Caesarean section under regional anaesthesia were randomly allocated to receive 2.0 ml hyperbaric bupivacaine 0.5% in either the sitting or left lateral position. Spinal injection was performed with a 27 gauge, 120 mm long spinal needle using a single space combined spinal extradural technique. The onset time to analgesia at T4, and grade 3 motor block was on average 7.7 min and 6.9 min respectively in the lateral group, compared with 10.8 min (P < 0.05) and 9.4 min (P < 0.05) in the sitting group. Nine women in the sitting group and one woman (P < 0.05) in the lateral group required epidural supplementation. Hypotension occurred in 48% of the parturients in the lateral group and in 13% (P < 0.05) of the parturients in the sitting group. Nausea was noted in 61% of the parturients in the lateral group and in 22% (P < 0.05) of the parturients in the sitting group. There was no difference between the two groups in neonatal outcome. Overall, the position of the patient during induction of spinal anaesthesia does influence the rate of onset of analgesia and motor blockade. Injection of 10 mg hyperbaric bupivacaine in the sitting position would not provide adequate analgesia for Caesarean section when using a single space combined spinal extradural technique.

Adult↗
PubMed1993

Cellular biology of human intimal hyperplastic stenosis.

Restenosis after angioplasty and vascular surgery remains a major unsolved clinical problem. Vascular smooth muscle cell (VSMC) hyperplasia is an invariable response, but in 20-50% of cases proceeds to compromise the vessel lumen. We sought to identify cellular characteristics of human VSMC which are associated with restenosis. Human VSMC were grown from 135 samples of vascular tissue derived from patients undergoing primary cardiovascular surgery and revision surgery for restenosis. Cells derived from normal vein and artery, atherosclerotic plaques and from stenotic lesions were studied for successful proliferation in cell culture. Furthermore, growth rates were measured in response to 15% foetal calf serum +/- inhibition with heparin (100 micrograms/ml). Significantly fewer cells from atherosclerotic plaques progress to the third passage in cell culture than those derived from stenoses and controls (p < 0.001, Chi square) and growth rates after the third passage could not be studied in these cells. Of cells that progress to this stage, growth rates do not differ between stenosis-derived and normal cells under standard conditions. VSMC from mature atherosclerotic plaques may have undergone senescent changes. Stenosis-derived cells do not grow more rapidly than normal cells, but are significantly less sensitive to heparin (p < 0.001, Mann-Witney test), which is a major physiological inhibitor of VSMC growth. Differences in biological characteristics of human VSMC, observed in cell culture, may provide important insights into human vascular disease.

Actins↗
PubMed1993

Cell-surface heparan sulfate proteoglycan mediates HIV-1 infection of T-cell lines.

The role of cell-surface proteoglycans in human immunodeficiency virus (HIV) infection of T-cell lines was investigated. HIV-1-susceptible lymphoblastic T-cell lines, MT-4 and H9, were analyzed for proteoglycan synthesis and found to make heparan sulfate (HS) and chondroitin sulfate proteoglycans. Enzymatic treatment of these cells with heparitinase, but not chondroitinase, significantly prevented HIV-1(IIIB) infection as measured by inhibition of cytopathicity, reverse transcriptase production, and syncytia formation. Sulfation of glycosaminoglycans HS chains was critical to viral entry as shown by inhibition of viral infection with sodium chlorate and its specific reversal with exogenous sulfate addition. Quantitation of direct virus binding to cells showed that treatment of cells with heparitinase inhibited HIV-1 binding to the T-cell surface. Exogenous HS added to cultures inhibited virus infection in a manner analogous to dextran sulfate, further supporting a functional role for HS in HIV-1 binding. These results provide evidence for participation of cell-surface HS proteoglycans in HIV-cell attachment and virus entry.

Binding Sites↗
PubMed1993

Depression and the experience of psychological deficits in schizophrenia.

A study of the relationships between depression and the subjective experience of psychological deficits was carried out in a group of 50 schizophrenic patients selected from a population of long-term mentally ill patients. Experience of psychological deficits was associated with depression, and furthermore the temporal relationships between the phenomena supported the hypothesis that experience of psychological deficits is associated with vulnerability to depression in schizophrenia. In addition, the patients' self-reporting of depressed mood and negative cognitions was congruent with an observer's assessment of depression. These findings indicate that subjective experiences of deficits characteristic of the schizophrenic illness confer vulnerability to depression, but nonetheless the patients' experience of depression resembles that typical of depressed non-schizophrenic patients.

Adult↗
PubMed1993

Identification and characterization of multiple species of vancomycin-resistant enterococci, including an evaluation of Vitek software version 7.1.

A total of 374 clinical isolates of Enterococcus spp. were characterized to determine the species distribution and vancomycin resistance. The ability of the Vitek system (bioMerieux Inc, Hazelwood, St. Louis, Mo.) to identify enterococci to the species level and to recognize vancomycin resistance by using computer software version 7.1 was evaluated. Conventional methods were used for identification and agar dilution was used for susceptibility testing, the results of which were as follows (presented as number of vancomycin-resistant isolates/number of members of that species identified): 219/234 E. faecium, 9/112 E. faecalis, 2/3 E. mundtii, 0/1 E. durans, 0/1 E. hirae, 0/1 E. raffinosis, and 0/1 E. avium. Ten enterococci were in the vancomycin-intermediate category (six E. gallinarum, two E. casseliflavus, and one each of E. faecium and E. faecalis). The Vitek GPI card correctly identified 98% of E. faecium isolates, 99% of E. faecalis isolates, and only two isolates of other enterococcal species. The GPS-TA card was 98% sensitive and 95% specific for the detection of vancomycin resistance, generating a total of four very major (1.6%) and five minor errors (1.3%).

Drug Resistance, Microbial↗
PubMed1993

Dietary n-3 fatty acid effects on neutrophil lipid composition and mediator production. Influence of duration and dosage.

Healthy volunteers supplemented their usual Western diets with Promega fish oil supplement (eicosapentaenoic acid [EPA], 0.28 g; docosahexaenoic acid [DCHA], 0.12 g; other n-3 fatty acids 0.10 g per capsule) using three protocols. Initial experiments (protocol 1 and 2) investigated the kinetics of incorporation of n-3 fatty acids into serum and neutrophil lipids after 10 capsules/d of Promega. EPA was rapidly detected in both serum and neutrophil lipids; the arachidonic acid (AA) to EPA ratio in neutrophil phospholipids showed a maximal reduction of 49:1 to 8:1 within 1 wk of beginning supplementation. EPA was preferentially incorporated into phosphatidyl-ethanolamine and phosphatidylcholine but not phosphatidylinositol. Long-term supplementation for up to 7 wk did not influence the AA/EPA ratio or the distribution of EPA among neutrophil phospholipids in a manner that was not observed after the first week. Neutrophils produced similar quantities of platelet-activating factor and slightly lower quantities of leukotriene B4 during long-term supplementation when compared with presupplementation values. Experiments examining the influence of Promega dosage indicated that the AA/EPA ratio in neutrophil lipids decreased in a dose-dependent manner. Only when the dose was increased to 15 capsules/d was there a reduction in the AA/DCHA ratio in neutrophil lipids. The quantity of AA in neutrophil lipids remained relatively constant at all supplement doses. Taken together, the current study demonstrates the capacity of n-3 fatty acids provided with a Western diet to be rapidly incorporated into neutrophil lipids. However, dietary n-3 fatty acids appear not to significantly reduce arachidonate content within neutrophil phospholipids. Constant arachidonate levels may account for the lack of large reductions in the biosynthesis of lipid mediators by neutrophils after fish-oil supplementation.

Adult↗