PubMed Health⌕ Search

Biomedical subjects

E Meezan

Publications and source records attributed to E Meezan.

At least 55 records · Page 3Linked to original sources
PubMed1985

Sulfate transport in human lung fibroblasts (IMR-90).

Sulfate transport in a fibroblast cell line derived from human lung (IMR-90) occurred mainly via high- and low-affinity, SITS-sensitive pathways and to a lesser extent by an SITS-insensitive mechanism. In low-ionic-strength media (sucrose substituted for salts) the apparent Km of the carrier-mediated sulfate influx was 1 mM. At 0.3 mM, the sulfate concentration normally found in human serum, the contribution of the SITS-insensitive pathway was negligible. In physiological salts solution, an SITS-sensitive, high-affinity (Km 34 +/- 14 microM) sulfate influx system was observed at extracellular sulfate concentrations less than 100 microM. Between 100 and 500 microM sulfate, the range normally found in human serum, sulfate influx occurred via an SITS-sensitive, low-affinity pathway and to a small extent by an SITS-insensitive mechanism. Extracellular chloride inhibited the influx and stimulated the efflux of sulfate. Bicarbonate and thiosulfate inhibited sulfate influx but had no effect on sulfate efflux. Phosphate, arsenate, or Na+ did not affect sulfate uptake. These results indicate that in human lung fibroblast IMR-90 cells sulfate is transported mainly via an SO4(2-)/Cl- exchange system independent of the phosphate or Na+ transport. Since sulfate concentration as high as 50 mM only slightly increased sulfate efflux, SO4(2-)/SO4(2-) exchange is probably a minor component of sulfate uptake.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗
PubMed1985

Identification of the insulin receptor of cerebral microvessels.

Cerebral microvessels are known to possess receptors for insulin and have recently been shown to respond to physiological levels of this hormone. Scatchard analysis of binding data obtained with isolated cerebral microvessels gave curvilinear plots and showed that neonatal porcine cerebral microvessels have a greater number of insulin receptors per unit of protein than adult bovine cerebral microvessels. The high-affinity form of the insulin receptors of both neonatal porcine and adult bovine cerebral microvessels have similar binding constants (dissociation constant = 0.3 X 10(-9) M). Dissociation of 125I-insulin from cerebral microvessels was accelerated by the presence of unlabeled insulin in preparations from both neonatal pigs and adult cows. 125I-insulin was covalently cross-linked to its receptor in cerebral microvessels with disuccinimidyl suberate, and the hormone-receptor complex was isolated on sodium dodecyl sulfate-polyacrylamide gels. Under reducing conditions, 125I-insulin was found associated with a polypeptide with a molecular weight of 130,000, which is indistinguishable from the alpha-subunit of the liver insulin receptor. In contrast, nonvascular cerebral cortical tissue contained an insulin receptor with an alpha-subunit that was lower in molecular weight than the form isolated from cerebral cortical microvessels.

Animals↗
PubMed1985

Liquid-chromatographic determination of chloride in sweat from cystic fibrosis patients and normal persons.

Chloride concentrations in sweat were measured by "vacancy liquid chromatography," a technique in which anions eluting from an ion-exchange column in the presence of an elution buffer which absorbs at 280 nm cause a decrease in absorbance proportional to the concentration of the anion. Less than 1 microL of sweat suffices for this analysis, which takes 5 min per sample. Results were comparable to those obtained by conventional titrimetric determination of chloride. Sweat from cystic fibrosis patients had higher (104 +/- 26 mmol/L) chloride concentrations than sweat from normal persons (16 +/- 7 mmol/L). This technique can be used in the diagnosis of cystic fibrosis.

Child, Preschool↗
PubMed1984

Polyamines stimulate D-glucose transport in isolated renal brush-border membrane vesicles.

Polyamines are natural constituents of most living organisms. However, their function in normal or pathological conditions is not fully understood. We have investigated in vitro effects of polyamines on characteristic properties of isolated renal brush-border membrane vesicles in order to determine whether polyamines have a regulatory role in membrane transport processes. The polyamines putrescine, spermidine and spermine were found to stimulate D-glucose uptake. Diffusional L-glucose uptake was not altered, indicating that the polyamines affected the active transport of D-glucose, rather than inducing nonspecific changes in membrane lipid properties. The amiloride-sensitive Na+ /H + exchange was slightly inhibited by polyamines while Mg2+ -ATPase activity was stimulated. The polyamine effects could not be explained solely by the polycationic properties of these agents, since polycationic polypeptides had an opposite effect. For example, lysozyme was found to inhibit D-glucose transport. Spermine was incorporated into the trichloroacetic acid-insoluble fraction of brush-border membrane proteins. Results indicated that this incorporation process consisted of at least two components: a Ca2+ -independent component and a Ca2+ -dependent component, possibly as a result of transglutaminase activity which was present in the isolated renal brush-border membranes. By using SDS-polyacrylamide gel electrophoresis in conjunction with fluorography, [3H]spermine was shown to be incorporated into several brush-border membrane proteins, mainly the 57 kDa, 74 kDa, 100 kDa, a heavy molecular weight band (greater than 200 kDa) and a low molecular weight band (less than 10 kDa). Our results suggest that the polyamine effects on membrane function may be due to a covalent modification of membrane proteins, possibly via a transglutaminase-mediated incorporation of polyamines or to the crosslinking of membrane proteins.

Adenosine Triphosphatases↗
PubMed1984

Serum sulfate levels in patients with cystic fibrosis.

Patients with cystic fibrosis (CF) secrete copious amounts of mucous material which is viscous, tends to accumulate in the respiratory tract and contains larger than normal amounts of sulfate. The present investigation was designed to measure sulfate levels in the serum of patients with cystic fibrosis by ion chromatography of protein-free serum aliquots. The level of inorganic sulfate in the serum of non-cystic fibrosis pediatric patients averaged 0.29 +/- 0.03 mmol/l while patients suffering from cystic fibrosis had an average serum sulfate value of 0.27 +/- 0.03 mmol/l which was not significantly different from controls. No differences were observed in serum sulfate levels among males and females of either group of patients. There was a tendency for serum sulfate levels to decrease with age, but there was no statistically significant difference in serum sulfate levels between cystic fibrosis patients and normals as a function of age. These findings indicate that the highly sulfated mucoid materials secreted by cystic fibrosis patients are not reflected in abnormal serum sulfate levels.

Adolescent↗
PubMed1984

Distribution of insulin receptors in human erythrocyte membranes. Insulin binding to sealed right-side-out and inside-out human erythrocyte vesicles.

Analyses of insulin binding to human erythrocytes and to resealed right-side-out and inside-out erythrocyte membrane vesicles have revealed that high affinity insulin binding receptors are present on both sides of the erythrocyte membranes. Insulin binding to human erythrocytes was examined with the use of a binding assay designed to minimize the potential errors arising from the low binding capacity of this cell type and from non-specific binding in the assay. Scatchard analysis of equilibrium binding to the cells revealed a class of high affinity sites with a dissociation constant (Kd) of (1.5 +/- 0.5) X 10(-8) M and a maximum binding capacity of 50 +/- 5 sites per cell. Interestingly, both resealed right-side-out and inside-out membrane vesicles exhibited nearly identical specific sites for insulin binding. At the high affinity binding sites, for both right-side-out and inside-out vesicles, the dissociation constant (Kd) was (1.5 +/- 0.5) X 10(-8) M, and the maximum binding capacity was 17 +/- 3 sites per cell equivalent. These findings suggest that insulin receptors are present on both sides of the plasma membrane and are consistent with the participation of the erythrocyte insulin receptors in an endocytic/recycling pathway which mediates receptor-ligand internalization/externalization.

Binding Sites↗
PubMed1984

Isolation and characterization of beta 1-adrenergic receptors from adult, rat cardiac myocytes.

The beta-receptors were isolated from rat cardiac myocytes and characterized. Isolated myocytes were prepared from adult rat hearts and characterized for viability. Membrane proteins were solubilized from myocytes with 1% Triton X-102. The solubilized membrane proteins were fractionated by DEAE-Sephacel ion exchange column chromatography. Two major protein peaks were obtained. The second protein peak sample was found to contain beta-receptors to which 125I-15-(4'-azido-3'-iodobenzyl)-carazolol (125I-ABC) was specifically bound. This sample was labeled covalently with 125I-ABC by UV irradiation. The radiolabeled sample was applied to a Sepharose CL-6B gel column. Two radiolabeled protein peaks, one with a molecular weight of approximately 570,000 and the other with a molecular weight of approximately 95,000 were found. When the 570,000-dalton complex was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, it was dissociated into a component with a molecular weight of 66,000. The 95,000-dalton complex was dissociated into a 58,000-dalton component upon SDS-PAGE under reducing conditions. An excess amount of isoproterenol and propranolol decreased photolabeling of the beta-receptors with 125I-ABC by 60% and 40%, respectively.

Adrenergic beta-Antagonists↗
PubMed1984

Identification and characterization of the insulin receptor of bovine retinal microvessels.

The presence of specific, high affinity receptors for insulin has been demonstrated in purified preparations of bovine retinal microvessels. The binding of [125I]insulin to isolated retinal microvessels was inhibited by unlabeled insulin, but not by other peptide hormones. Scatchard analysis of the [125I]insulin binding data gave a curvilinear plot similar to that exhibited by insulin receptors in known insulin-sensitive tissues such as adipocytes and hepatocytes. Binding of [125I]insulin to retinal microvessels, followed by covalent cross-linking of the bound ligand to the alpha-subunit of the insulin receptor with the bifunctional reagent disuccinimidyl suberate, yielded a prominent specific [125I]insulin-labeled band when analyzed by sodium dodecyl sulfate-gel electrophoresis followed by autoradiography, and this band had a mobility identical to that of the corresponding complex obtained with rat liver plasma membranes (mol wt, 125,000). These results demonstrate for the first time that the retinal microvasculature, a major site of pathological injury in diabetes mellitus, contains insulin receptors that are similar to those present in known insulin-sensitive tissues, such as liver, fat, and muscle.

Animals↗
PubMed1984

Increased renal tubular sodium pump and Na+,K+-adenosine triphosphatase in streptozotocin-diabetic rats.

The effects of chronic glucose osmotic diuresis on renal tubular sodium pump and Na+,K+-adenosine triphosphatase (ATPase) activities were studied in chronic streptozotocin-induced diabetic rats. Four to seven weeks after streptozotocin (60 mg/kg i.p.) injection, specific renal Na+,K+-ATPase activity showed a 34.8% increase as compared to the saline-citrate treated controls, whereas the nonspecific Mg++-ATPase was not altered. The concentration of Na+,K+-ATPase, estimated from the maximum [3H]ouabain binding site concentration, also showed a significant increase in the chronic streptozotocin-diabetic rats. To determine further the specificity of this increase in Na+,K+-ATPase, the activity of the sodium pump, estimated from ouabain-sensitive 86Rb uptake, was measured in nonenzymatically isolated renal tubules. Again, a significant (+106.4%) increase in the renal tubular sodium pump activity was observed in the streptozotocin-diabetic rats, whereas the nonspecific, ouabain-insensitive 86Rb uptake was not altered. Neither was there any difference in 86Rb uptake by the isolated renal glomeruli. Thus, it appears that chronic streptozotocin-induced diabetes in rats is associated with a significant increase in renal tubular sodium pump and Na+,K+-ATPase. The latter effects may represent an important physiologic adaptation of the kidneys to maintain electrolyte homeostasis in diabetes.

Adenosine Triphosphatases↗
PubMed1983

Isolation and characterization of human erythrocyte insulin receptors.

The noncovalently associated 125I-insulin-receptor complex was isolated from human erythrocyte membranes after allowing 125I-insulin to interact with the membranes followed by extraction of the 125I-insulin-receptor complex with Triton X-102 or, alternatively, by complete solubilization of the membranes with sodium dodecyl sulfate (SDS), removal of SDS, and then treatment of the solubilized sample with 125I-insulin. Sepharose CL-6B column chromatography of the 125I-insulin-receptor complex obtained by both of the above procedures yielded a highly radioactive 140,000-Da complex which was dissociated into small peptides when subjected to SDS-polyacrylamide gel electrophoresis. In contrast, when the 125I-insulin-treated membrane sample was extracted with Triton X-102, purified by DEAE-Sephacel ion exchange chromatography, covalently cross-linked with disuccinimidyl suberate, and then subjected to SDS-polyacrylamide gel electrophoresis, a highly radioactive component with Mr = 53,000 was obtained. On the other hand, when the Triton X-102-solubilized membrane receptor sample was fractionated by DEAE-Sephacel ion exchange chromatography, complexed with 125I-insulin, covalently crosslinked, and then applied to a Sepharose CL-6B column, a 95,000-Da complex with high specific radioactivity was obtained. Upon SDS-polyacrylamide gel electrophoresis, the 95,000-Da complex was dissociated into a 53,000-Da component which appeared identical with that obtained from the receptor complex described above which was obtained by direct interaction of the membranes with 125I-insulin.

Chromatography, Gel↗
PubMed1983

Selective solubilization of two populations of polypeptides from bovine retinal basement membranes.

Basement membranes isolated from the entire neural retinae of normal cows consist of a heterogeneous mixture of polypeptides resolvable as two major populations, one collagenous and the other non-collagenous. The non-collagenous population consists of disulfide-bonded aggregates of which the major fraction is soluble in lithium dodecyl sulfate (LDS) at 4 degrees C without reducing agent, with the remainder requiring reduction for solubilization at 4 degrees C. Only the second 4 degrees C extract is contaminated with small amounts of hydroxylated amino acids. The non-collagenous population consists of up to 25 polypeptides detectable in sodium dodecyl sulfate (SDS) gels, all of which stain blue with Coomassie Blue R-250, are insensitive to bacterial collagenase and migrate on or slightly below the central diagonal in urea-SDS/SDS electrophoresis. The collagenous population is soluble in LDS at 100 degrees C under reducing conditions and has a collagenous amino acid composition, although consisting of only 272 glycine residues/1000. This fraction consists of four major and four minor polypeptides detectable in SDS gels, all of which stain red (metachromatically) with Coomassie Blue R-250, are degraded by bacterial collagenase and migrate below the central diagonal in urea-SDS/SDS electrophoresis. The four major collagenous polypeptides are larger than the alpha-chains of type I collagen. In total, the solubilized proteins account for 60% of the general protein, but only 30% of the sum of hydroxyproline and hydroxylysine. The insoluble residue has a collagenous amino acid composition similar to that of the 100 degrees C extract, but with lower 3-hydroxyproline and hydroxylysine contents.

Amino Acids↗
PubMed1983

Rapid isolation of type II pneumocytes with magnetic removal of macrophages.

A method is described for the rapid preparation of lung cell fractions enriched in type II alveolar pneumocytes. Isolated perfused rabbit lungs are exposed to Fe3O4 by tracheal lavage, which permits pulmonary alveolar macrophages to phagocytize the particles. Alveolar epithelial cells are then selectively freed from the basement membrane matrix by critical placement of collagenase and elastase. Detached cells are harvested either by repeated tracheal lavage or by mincing the lobes and filtering freed cells through a series of nylon mesh sieves. Iron oxide-containing macrophages are then removed from the harvested cells by a strong magnetic field. A final sizing of the macrophage-depleted suspension yields a preparation enriched in alveolar type II cells. Eight million viable cells (95% type II) were obtained per rabbit lung when harvested by lavage, while 32 x 10(6) (88% type II) cells were obtained from minced lungs. These values for cell yield and relative purity are comparable to previously described separation methods that depend upon differences in cell density or size. A major advantage of the magnetic separation procedure is the substantially shortened preparation time, typically 2 hr instead of 4. The viability (90-95%), oxygen consumption (88 nmol/10(6) cells/hr), and incorporation of [14C]acetate and [14C]choline (0.44 and 0.115 nmol/10(6) cells/hr, respectively) indicate that these cells will be suitable for pharmacologic and toxicologic investigation.

Animals↗
PubMed1982

Partial characterization of insulin receptors from rat myocytes.

Isolated myocytes were prepared from the adult rat heart and characterized for viability. The myocytes were exposed to 125I-insulin, and the 125I-insulin-receptor complex was extracted with 1% Triton X-102 and then applied to a DEAE-Sephacel column. When the chromatography were applied to a Sepharose CL-6B column, a 140,000-dalton complex with high specific radioactivity was found. Alternatively, when myocyte insulin receptors were first extracted with 1% Triton X-102 without prior exposure to 125I-insulin and then applied to a DEAE-Sephacel column, three peak protein fractions were obtained. They were treated separately with 125I-insulin and the 125I-insulin-protein complexes were covalently cross-linked with disuccinimidyl suberate. The cross-linked samples were applied to a Sepharose CL-6B column and the radioactive protein fractions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The first and second peak samples from the ion exchange chromatography yielded complexes of low specific radioactivity, which appear to be formed by nonspecific random binding of 125I-insulin to the solubilized membrane proteins. In contrast, Sepharose CL-6B gel filtration of the cross-linked sample from the third peak fractions gave a major highly radioactive 125I-insulin-receptor complex with a molecular weight of 370,000 and a minor complex of low radioactivity with a molecular weight of 140,000. Upon sodium dodecyl sulfate-gel electrophoresis, the 370,000-dalton complex was dissociated to 130,000- and 82,000-dalton components and the 140,000-dalton complex was dissociated to a 47,000-dalton component.

Animals↗
PubMed1982

Preparation of cell-free extracellular matrix from human peripheral nerve.

The extracellular matrix of human peripheral nerve, which is mainly basement membrane and fibrillar collagen, has been prepared by a procedure involving extensive detergent extraction of isolated endoneurium and perineurium obtained from various nerves. The ultrastructure of the isolated nerve extracellular matrix was indistinguishable from that seen in sections of intact nerve, indicating that the extraction procedure preserved the morphological integrity of these connective tissue components. The amino acid and carbohydrate compositions of the nerve extracellular matrix preparations were typically collagenous in nature containing a high content of glycine, proline, 4-hydroxyproline, and alanine and significant amounts of lysine and hydroxylysine. The preparations contained virtually no 3-hydroxyproline and low content of glucose and galactose compared to pure basement membranes, indicating that interstitial rather than basement membrane collagens predominated. This preparation appears well-suited to both the ultrastructural and biochemical study of the extracellular matrix of peripheral nerve.

Cell Fractionation↗
PubMed1982

Thickened cerebral cortical capillary basement membranes in diabetics.

In human diabetes mellitus, thickening of the microvascular endothelial cell basement membrane has been documented by quantitative ultrastructural studies in a variety of vascular beds. We studied the thickness of the cerebral cortical capillary basement membrane in frontal and occipital lobes from 21 diabetics and 16 nondiabetics, and found that those from diabetics were significantly thicker. Skeletal muscle capillary basement membranes have been shown to thicken with increasing age. However, only in the frontal cortex of diabetics was a significant relationship observed between capillary basement membrane thickening and increasing age. We conclude that, along with many of the previously studied microvascular beds, the cerebral cortical microvasculature displays thickening of the endothelial cell basement membrane in diabetes mellitus. No currently known disease is associated with this microvascular change, unlike the closely related retinal vasculature, where the association with diabetic retinopathy is established.

Adult↗