Cell adhesion. Mucins in the mainstream.
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Cell Adhesion↗
Biomedical subjects
S Shaw
Publications and source records attributed to S Shaw.
A high-performance liquid chromatographic assay for reduced and oxidized glutathione in biological samples.
A high-performance liquid chromatographic method was developed for the measurement of oxidized and reduced glutathione in biological samples. The method allowed the separation of glutathione (oxidized and reduced) from related thiols (homocysteine, cysteine, cystine, methionine) and other intermediates of glutathione pathways without derivatization or enzymatic reduction. Quantitation was achieved with uv detection. Biological samples were prepared by rapid homogenization in iced KCl followed immediately by deproteinization and acidification with sulfosalicylic acid. Samples were eluted isocratically at 1 ml/min using 0.0025 M sodium phosphate buffer, pH 3.50, containing 0.005 M tetrabutylammonium phosphate (Waters, Milford, MA) and 13% methanol and analyzed on a 30-cm x 3.9-mm C-18 mu Bondapak column and detected with a uv detector at 190 nm. The determination of nanomole levels of glutathione and glutathione disulfide and their separation from other thiols are described.
Animals↗
Selective expression of integrin alpha 4 beta 7 on a subset of human CD4+ memory T cells with Hallmarks of gut-trophism.
Human memory CD4+ T lymphocytes are heterogenous in expression of integrins; one subset has the unexpected phenotype beta 1 low alpha 4 high. We demonstrate that this subset is unique among CD4+ cells in expression of high levels of alpha 4 beta 7, detected by a distinctive mAb Act-1. alpha 4 beta 7 is involved in binding to both fibronectin and vascular cell adhesion molecule-1; Act-1 blocks cell binding to the former and augments binding to the latter. Act-1 expression marks a subset of memory cells that, unlike the predominant circulating memory cell, has up-regulated beta 7 rather than beta 1. Their phenotype is distinct from that described for skin-homing T cells and is fully consistent with that described for gut-homing T cells. Differential adhesion capacity of this subset is verified by selective binding to FN and vascular cell adhesion molecule-1 in a beta 1-independent fashion. Thus, alpha 4 beta 7 detected on this subset of circulating normal T cells fits the expectations for a gut-homing receptor.
Antibodies, Monoclonal↗
Insulin sensitivity and body fat distribution in normotensive offspring of hypertensive parents.
Lean, healthy normotensive sons of essential hypertensive parents (OHyp) have lower insulin sensitivity (SI) than sons of normotensive parents (ONorm). We have tried to find out whether this disturbance in insulin metabolism is related to altered body fat distribution, fuel metabolism, or both. 21 OHyp and 21 ONorm of similar age and body-mass index were investigated after fasting overnight. Body composition was assessed by dual-energy X-ray absorptiometry and fuel metabolism by indirect calorimetry and urinary nitrogen excretion. Plasma insulin and glucose concentrations were measured during the frequent sampling intravenous glucose tolerance test, and SI was calculated by the minimum model method. Systolic blood pressure and heart rate were slightly but not significantly higher in OHyp than ONorm but the groups did not differ in fasting plasma insulin or glucose concentrations, carbohydrate or lipid oxidation, lean and fat mass, bone mineral content, or distribution of body fat. By contrast, SI was significantly lower in OHyp than ONorm (8.2 [0.7] vs 13.4 [1.5] 10(-4) L mU-1 min-1, p < 0.01). Within the whole study population upper-body fat mass was positively correlated with fasting plasma insulin (r = 0.33, p < 0.03) and lipid oxidation was positively correlated with SI (r = 0.35, p < 0.04) and negatively correlated with subscapular/triceps skinfold thickness (r = -0.43, p < 0.01). Thus, impairment of SI precedes both the development of overt hypertension and gain or redistribution of body fat. Therefore, the concept that SI is low as a result of altered fat distribution has to be reconsidered, at least in young male offspring of hypertensive parents.
Adipose Tissue↗
T-cell adhesion induced by proteoglycan-immobilized cytokine MIP-1 beta.
Lymphocyte migration from blood into tissue depends on integrin-mediated adhesion to endothelium. Adhesion requires not only integrin ligands on the endothelium, but also activation signals because T-cell integrins cannot bind well until they are activated. The physiological 'triggers' for T-cell adhesion are unknown, but cytokines may be good candidates as they are released during inflammation and trigger adhesion in neutrophils and monocytes. We have identified a cytokine, macrophage inflammatory protein-1 beta (MIP-1 beta), that induces both chemotaxis and adhesion of T cells; MIP-1 beta is most effective at augmenting adhesion of CD8+ T cells to the vascular cell adhesion molecule VCAM-1. We reasoned that, as cytokines in vivo will be rapidly washed away, MIP-1 beta might be bound to endothelial surfaces and so induce adhesion in its immobilized form. Here we show that: (1) MIP-1 beta is present on lymph node endothelium; (2) immobilized MIP-1 beta induces binding of T cells to VCAM-1 in vitro. MIP-1 beta was immobilized by binding to proteoglycan: a conjugate of heparin with bovine serum albumin and cellular proteoglycan CD44 were both effective. We propose that MIP-1 beta and other cytokines with glycosaminoglycan-binding sites will bind to and be presented by endothelial proteoglycans to trigger adhesion selectively not only of lymphocyte subsets, but also of other cell types.
Cell Adhesion↗
T cell adhesion to endothelium: the FRC conduit system and other anatomic and molecular features which facilitate the adhesion cascade in lymph node.
Since T cell surveillance depends on movement from blood into tissue and back again, rapid, efficient and selective T cell adhesion to vascular endothelium is essential. This adhesion involves a multistep cascade clarified by a recent consensus model: (1) initial tethering by selectin-mediated interactions; (2) triggering of adhesive function of T cell integrins by ligands at or near the endothelial surface; and (3) strong adhesion mediated by T cell integrins. We recapitulate this model, particularly as it pertains to the lymph node, and explore additional molecular and anatomic elements which contribute to the effectiveness of the adhesion cascades at that site: (1) importance of cytokines/soluble mediators as triggering ligands; (2) role of glycocalyx and proteoglycans on high endothelial venule (HEV) endothelium in capturing and presenting triggering cytokines; (3) remarkable function of what we designate the 'fibroblastic reticular cell (FRC) conduit system' in rapidly transporting cytokines to the HEV; (4) importance of the unique anatomy of the flap-valve junctions between HEV endothelium in enabling intravasation of cytokines and transmigration of lymphocytes. Taken together, these molecular mechanisms and these three anatomic features of lymph node facilitate extremely efficient lymphocyte traffic to this site critical for T cell-mediated immune responses. Analogous mechanisms contribute to T cell interaction with endothelium at other sites.
Animals↗
Regulation of leukocyte recruitment by proadhesive cytokines immobilized on endothelial proteoglycan.
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Animals↗
Lack of effect of long-term amlodipine on insulin sensitivity and plasma insulin in obese patients with essential hypertension.
To evaluate the effects of long-term treatment antihypertensive with the dihydropyridine calcium antagonist amlodipine on insulin sensitivity, plasma insulin, and lipoprotein metabolism in obese hypertensive patients. We measured the insulin sensitivity index (SI), determined by the Minimal Model Method of Bergman, fasting plasma insulin and glucose concentrations, serum total triglyceride and lipoprotein cholesterol fractions, and blood pressure in 20 obese, non-diabetic patients with essential hypertension before and after 6 weeks of placebo and again after 6 months of amlodipine. Ten patients [mean body mass index (BMI) 30.2 kg.m-2] had been on prior treatment with a thiazide diuretic in low dosage and/or a beta-adrenoceptor blocker (group A), and 10 matched patients [BMI 31.8 kg.m-2] had been previously untreated (group B). Amlodipine was started in a dose of 5 mg and was increased to 10 mg once daily in 14 patients who were hypertensive after 8 weeks on the lower dosage. At entry (before placebo), SI was slightly but not significantly lower in group A than B [2.7 vs. 3.6 x 10(-4) ml.microU-4.min-1]; fasting plasma insulin was 13.6 vs. 12.9 microU.ml-1. After 6 weeks on placebo, S1 averaged 3.7 in group A and 4.4 x 10(-4) microU.ml-1.min-1 in group B; fasting plasma insulin was 14.6 vs. 15.1 microU.ml-1, and glucose 5.5 vs. 5.5 mmol.l-1.(ABSTRACT TRUNCATED AT 250 WORDS)
Amlodipine↗
Specificity effects of run versus cycle training on ventilatory threshold.
This study compared the effects of 9 weeks of run (RT) versus cycle (CT) training on ventilatory threshold (Thv) determined during treadmill (TM) and cycle ergometer (CE) graded exercise testing. Sixteen college age men were assigned to a RT or CT group and performed a TM and a CE test before and after training. Both training groups performed similar training protocols which initially consisted of continuous exercise 4 days.week-1 at 75-80% maximum heart rate (fc, max) for 45 min. Training intensity was later increased to 80-85% fc, max and interval training (90-95% fc, max) was incorporated 2 days.week-1 into the continuous training. Both groups showed significantly improved maximal oxygen consumption (VO2 max) on both TM and CE tests (P < 0.01) with no significant differences between the groups. Significant Thv increases (P < 0.05) were found on TM tests for RT (n = 8) and CT (n = 8) groups [mean (SD); 443 (438) and 373 (568) ml O2 x min-1, respectively] with no difference between the groups. Results from the CE tests revealed a significant Thv increase (P < 0.01) for the CT group [566 (663) ml O2 x min-1] with no change for the RT group. The Thv improvement noted for the RT group was significantly different (P < 0.05) comparing CE with TM tests but not for the CT group. The results indicate that CT and RT improvement in Thv for runners is dependent upon mode of training and testing, and there is an apparent dissociation of VO2 max and Thv specific to training.
Adult↗
Proteoglycans on endothelial cells present adhesion-inducing cytokines to leukocytes.
Leukocyte recruitment from the blood circulation into tissue is essential for effective immune responses, and is, consequently, carefully regulated. In this article Yoshiya Tanaka and co-workers describe a model in which proteoglycans on the luminal surface of endothelium capture pro-adhesive cytokines. These cytokines provide the adhesion-inducing signal to particular leukocyte subsets which initiates their transmigration.
Animals↗
HLA class II alleles and susceptibility and resistance to insulin dependent diabetes mellitus in Mexican-American families.
The role of HLA class II alleles in genetic predisposition to insulin dependent diabetes mellitus (IDDM) was examined by PCR/oligonucleotide probe typing of 42 Mexican-American IDDM families derived from Hispanic Caucasians and Native Americans. All high risk haplotypes (HLA-DR3 and DR4) were of European origin while the most strongly protective haplotype (DRB1*1402) was Native American. Of the 16 DR-DQ DR4 haplotypes identified, only those bearing DQB1*0302 conferred risk; the DRB1 allele, however, also markedly influenced IDDM risk. The general pattern of neutral and protective haplotypes indicates that the presence of Asp-57 in the HLA-DQ beta chain does not confer IDDM protection per se and indicates that both DRB1 and DQB1 influence IDDM susceptibility as well as protection.
Alleles↗
Analysis of HLA-DQA1 and -DQB1 genes in Mexican Americans with insulin-dependent diabetes mellitus.
Mexican American patients (n = 35) with insulin-dependent diabetes mellitus (IDDM) and control subjects (n = 39) were HLA-DQA and DQB typed by the polymerase chain reaction technique combined with allele-specific oligonucleotide probes. Either DQB1*0302 or DQB1*0201 was present among 91% (32/35) of the patients compared to 67% (26/39) of controls. Either DQA1*0501 or DQA1*0301 was present in all patients (100% or 35/35) compared to 29/39 (74%) (OR 12.06 Pc < 0.05) of controls. All four of these genes, in cis or trans, were present in 15/35 (43%) of the patients compared to 3/39 (8%) of controls (OR 9.0; Pc < 0.01). The presence of one or more non-susceptibility alleles showed a dose-related decrease in relative risk. Presence of aspartic acid (Asp) at position 57 of the DQ beta chain did not confer protection and non-Asp homozygosity did not confer susceptibility to IDDM in this ethnic group. In conclusion, susceptibility to IDDM in Mexican Americans is associated with particular DQA and DQB combinations, illustrates dose-dependent parameters and contradicts the critical residue hypothesis.
Adult↗
Allergic reactions to a hairdressers' series: results from 9 European centres. The European Environmental and Contact Dermatitis Research Group (EECDRG).
To obtain data on the frequency of sensitization among European hairdressers, the patch test results from 9 centres were evaluated. 8 allergens recommended by the ICDRG and EECDRG in the hairdressing series and PPD from the standard series were used to patch test 809 hairdressers and 104 clients suspected of contact sensitization. Among hairdressers, the mean frequencies of sensitization ranked as follows: GMT 19%, PPD 15%, APS 8%, PTD 8%, ONPPD 4% and PADH 4%. In contrast to GMT in acid permanent waves, the frequency of sensitization to AMT in alkaline permanent waves was only 4%. Frequencies of sensitization to pyrogallol and resorcinol were 0.8% and 0.6%, respectively. The frequencies of sensitization showed marked regional variations, particularly that to GMT, which was highest in Germany (51%), followed by Spain (22%) and London (19%). Clients of hairdressers showed a similar rank order of sensitization frequency, with the exception of APS, which was completely negative in this (small) series.
Beauty Culture↗
The pathogenesis of hypertension in obese subjects.
Obese subjects are at an increased risk of becoming hypertensive and vice versa. Essential hypertension and obesity are commonly accompanied by insulin resistance (defined as impaired insulin-mediated glucose disposal) and hyperinsulinaemia. In the offspring of patients with essential hypertension, insulin resistance and hyperinsulinaemia, as well as related increases in serum low density lipoproteins and triglycerides, often occur prior to the development of essential hypertension, overweight or central redistribution of body fat. Moreover, once obesity, and in particular central obesity, is present, insulin resistance is more marked in hypertensive than in normotensive obese subjects. Hyperinsulinaemia and/or insulin resistance in turn promote body fat deposition and impaired glucose tolerance. This cycle helps to explain why a familial predisposition to essential hypertension poses an increased risk of developing not only hypertension but also dyslipidaemia, obesity and non-insulin-dependent (type 2) diabetes. It is still unclear whether insulin resistance and/or hyperinsulinaemia also promote hypertension per se. Regardless of insulin's exact pathogenic role, obesity and/or a high dietary intake of carbohydrates, salt, etc. can induce several potential pressor mechanisms: 1) higher plasma noradrenaline (norepinephrine) and adrenaline (epinephrine) levels, suggesting a higher sympathetic tone in obese than in nonobese subjects, and in hypertensive obese than in normotensive obese subjects; 2) similarly, a tendency to hyperaldosteronism, with largely normal plasma renin activity, in obese hypertensive patients; 3) enhanced sensitivity of blood pressure to salt; 4) increased total blood volume (although it is normal relative to body surface area), leading to increased cardiac output and eventually eccentric left ventricular hypertrophy; and 5) increased cytosolic free Ca++ levels and reduced intracellular Mg++ levels in the blood cells of obese hypertensive patients and patients with non-insulin-dependent diabetes, although this finding cannot necessarily be extrapolated to cationic levels in vascular muscle cells. Total peripheral vascular resistance is usually low in normotensive obese subjects and rises with the development of hypertension; compared with lean patients with essential hypertension, obese hypertensive patients tend to have a slightly lower level of total peripheral vasoconstriction and a slightly higher cardiac output. Considering the intimate association between essential hypertension and obesity, as well as the prevalence and prognostic relevance of this combination, the spectrum of accompanying metabolic and cardiovascular abnormalities deserves careful consideration in the evaluation of therapeutic care for such patients.
Humans↗
Effects of stair-climbing vs run training on treadmill and track running performance.
Physically active college age women were evaluated to determine the effects of 9 wk of stair-climbing (Stairmaster Gauntlet) vs run training on 2414-m run time and treadmill measured aerobic capacity (VO2max) and submaximal physiological parameters. Subjects were randomly assigned to a stair-climbing (STAIR N = 11) (43.8 +/- 1.5 ml.kg-1.min-1) (mean +/- SEM) or run training (RUN N = 12) (44.2 +/- 1.8) group, training 4 d.wk-1 at 70-80% of maximum heart rate (MHR) for 30 min progressing to 85-90% MHR for 45 min. The STAIR group significantly increased (P < 0.01) their VO2max by 12% and decreased (P < 0.01) their 2414-m run time of 12.8 min by 8%. The RUN group increased (P < 0.01) their VO2max 16% and decreased run time (P < 0.01) 11% from 13.1 min. Submaximal treadmill runs at the same speed and grade demonstrated significant decreases in %VO2max and % MHR (P < 0.01) for both groups. The data support the use of stair-climbing exercise as an alternative mode to running with similar treadmill and running performance results subsequent to 9 wk of training.
Adult↗
Regulation of T-cell adhesion to endothelium.
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Cell Adhesion↗
[Does drinking protect against mountain sickness?].
This paper summarizes the main findings of 3 publications of our group [2-4] examining fluid balance at high altitude. Of 57 mountaineers ascending from 1170 m to 4559 m within 22 to 77 hours, 24 developed acute mountain sickness (AMS) and 16 developed high altitude pulmonary edema (HAPE). In 14 cases HAPE was preceded by symptoms of AMS. Independently of the amount of fluid intake, which varied from 2 to 4 l/24 h in these studies, subjects developing AMS showed decreased diuresis and natriuresis compared to healthy controls with similar fluid intake. Higher fluid intake resulted in greater urine output but did not prevent AMS. Higher plasma levels of aldosterone at rest and greater exercise-induced rises of plasma aldosterone and vasopressine may explain the increased water and salt retention in subjects with AMS. Whether these hormonal changes are secondary to a more severe hypoxemic stress or present a primary cause of AMS remains to be determined.
Adult↗
Costimulation of T cell receptor/CD3-mediated activation of resting human CD4+ T cells by leukocyte function-associated antigen-1 ligand intercellular cell adhesion molecule-1 involves prolonged inositol phospholipid hydrolysis and sustained increase of intracellular Ca2+ levels.
Activation of resting human CD4+ T cells mediated by mAb ligation of the TCR/CD3 complex requires costimulatory signals to result in proliferation; these can be provided by intercellular cell adhesion molecule-1 (ICAM-1, CD54) a natural ligand of leukocyte function-associated Ag-1 (LFA-1, CD11a/CD18). We analyzed early signaling events involved in T cell activation to determine the contribution by the LFA-1/ICAM-1 interaction. We studied in detail the hydrolysis of phosphatidylinositol(4,5)bisphosphate and intracellular levels of free Ca2+ during stimulation with beads coated with the CD3 mAb OKT3 alone or in combination with purified ICAM-1 protein. Our investigations show no response to LFA-1/ICAM-1 alone, but that costimulation by LFA-1/CAM-1 interaction induces prolonged inositol phospholipid hydrolysis (up to 4 h), resulting in generation of both inositol(1,4,5)phosphate3 and inositol(1,3,4,5)phosphate4 and their derivatives. Based on studies with cycloheximide, this costimulatory effect of prolonged inositol phospholipid hydrolysis appears dependent in part on de novo protein synthesis. A sustained increase in intracellular levels of free Ca2+ level is also observed after LFA-1/ICAM-1 costimulation, which is at least partly dependent on extracellular sources of Ca2+. Kinetic studies indicate that costimulation requires a minimal period of 4 h of LFA-1/ICAM-1 interaction to provide maximal costimulation for OKT3-mediated T cell proliferation. Thus, the necessary costimulation required for OKT3-mediated proliferation in this model system may be provided by an extended LFA-1/ICAM-1 interaction that in combination with OKT3 mAb leads to signal-transducing events, resulting in prolonged phospholipase C activation and phosphatidylinositol(4,5)bisphosphate hydrolysis, and a sustained increase in intracellular levels of free Ca2+.
CD4-Positive T-Lymphocytes↗