Interactions between insulin and its receptors after the initial binding event. Functional heterogeneity and relationships to insulin degradation.
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Adipose Tissue↗
Biomedical subjects
H Chang
Publications and source records attributed to H Chang.
Laser Raman studies on cobrotoxin.
Laser Raman spectra of cobrotoxin under various conditions have been obtained. Comparison of the spectra of native cobrotoxin in lyophilized form and in aqueous solution indicates that the secondary structures of cobrotoxin are not significantly affected by the removal of the aqueous solvent. On going from the native to the partially reduced and the completely reduced, carboxy-methylated forms, characteristic peaks of the C-S-S-C and tyrosine ring in the region of 500--900 cm-1 showed definite changes in structure. The partially reduced form gave two peaks at 502 and 524 cm-1, suggesting difference in the conformation of the remaining disulfide bonds. As indicated by the present work, the conformation of the main chain of cobrotoxin in the native unperturbed state, in the partially reduced and in the completely reduced forms are the coexistence of beta-pleated sheet with random-coil structure, predominantly random coil, and predominantly random coil with the existence of an alpha-helix type structure, respectively. The effect of pH on the conformation of cobrotoxin in solution appeared to give rise to the change of the local structure of two aromatic residues common to all snake neurotoxins.
Animals↗
Stimulated DNA synthesis in frog nuclei by cytoplasmic extracts of temperature-sensitive mammalian cells.
Cytoplasmic extracts of proliferating cells stimulate DNA synthesis in isolated nuclei of Xenopus laevis liver. When tested by the same assay, cytoplasmic extracts of resting cells are completely inactive. When cytoplasmic extracts are prepared from cell cycle-specific temperature-sensitive mutants arrestd in the G1 phase of the cell cycle by the nonpermissive temperature, they also fail to stimulate DNA synthesis in frog nuclei. The results indicate that, to stimulate DNA synthesis in isolated frog nuclei, essentially all information of G1 cells must be present.
Animals↗
Quantitation of hydrogen ion and potential gradients in gastric plasma membrane vesicles.
The ATP-dependent uptake of H+ by hog gastric parietal cell vesicles was quantitated by using the pH indicator dyes bromcresol green and malachite green, the weak bases, aminopyrine and 9-aminoacridine, and the pH electrode. A K+-dependent H+ uptake was found, with a significant difference between the quantity of H+ disappearing from the medium (deltaHo) and the quantity appearing inside the vesicle (deltaHi). 9-Aminoacridine gave a lower value for the deltaHi than any of the other probes. Probes of potential such as diethyloxadicarbocyanine or oxonol dyes showed that only secondary diffusion potentials occurred during H+ uptake and that the cationic dyes in the presence of protonophores could also be used to quantitate H+ uptake. The potential in the presence of protonophore indicated a deltaHi greater than that found with the other probes. Binding sites for acridine orange were generated either by ATP or an artificial pH gradient and corresponded to the deltaHi indicated by aminopyrine. SCN- (30mM) only partially inhibited the H+ gradient, and this, coupled with the failure to detect the physiological deltapH of 6.6, indicated that these vesicles may be an incomplete model of gastric acid secretion.
Adenosine Triphosphate↗
Chelation therapy in beta-thalassemia major. I. Intravenous and subcutaneous deferoxamine.
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Adolescent↗
Insulin binding to adipocytes. Evidence for functionally distinct receptors.
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Adipose Tissue↗
Measurement of sickling by controlled temperature increase.
Previous experiments to study the rate of red cell sickling have employed rapid mixing apparatus for SS cells with dithionite and have shown that the half-time of sickling is quite rapid, on the order of seconds. An alternative approach is to slow down the rate by taking advantage of the negative temperature coefficient of the process. We developed a method in which deoxygenation of a cell suspension is carried out at 0 degrees C. A linear temperature gradient to 37 degrees C is applied, and a gradual increase in the percentage of sickled cells is observed. At a heating rate of 1.5 degrees C/min the temperature at which half of the cells became sickled was 19 degrees C for SS cells treated with dithionite, 22 degrees C for SC cells, 28 degrees C for AS cells, 22 degrees C for cyanate-treated SS cells, and 23 degrees C for SS cells in the presence of 0.1 M butylurea. Thus this method promises to be useful for the study of sickling rates and the screening of potential anti-sickling agents.
Anemia, Sickle Cell↗
Conformational prediction for snake venom toxins and laser Raman scattering of a cardiotoxin from Taiwan cobra (Naja naja atra) venom.
Secondary structure regions in snake venom toxins were predicted using the prediction method of Chou and Fasman (Chou, P. Y., and Fasman, G. D. (1974), biochemistry 13,222) and an averaging scheme assuming structural homology in each type of toxins. The results indicate that, in general, snake toxins contain only some beta-sheet regions and beta bends. The content of secondary structures thus predicted does vary to some extent. The predicted results correlate well with conclusions from physicochemical studies. Interestingly, beta-bend regions predicted for the two types of neurotoxins, short-neurotoxin-type and long-neurotoxin-type, are primarily located in the middle of disulfide loops in spite of large differences in primary sequences. Comparisons between predicted results and the crystal structure of erabutoxin b determined at 2.75 A resolution suggest that the two types of neurotoxin are both sequencely and conformationally related while cardiotoxins could have an entirely different molecular topology. The Raman spectrum of a Taiwan cobra cardiotoxin indicates that the content of beta-pleated-sheet structure could be greater than that in neurotoxins.
Amino Acid Sequence↗
Proton transport by gastric membrane vesicles.
A highly purified membrane fraction was derived from hog gastric mucosa by a combination of differential and density gradient centrifugation and free flow electrophoresis. This final fraction was 35-fold enriched with respect to cation activated ouabain-insensitive ATPase. Antibody against this fraction was shown to be bound to the luminal surface of the gastric glands. The addition of ATP to this fraction or the density gradient fraction resulted in H+ uptake into an osmotically sensitive space. The apparent Km for ATP was 1.7-10(-4) M in the absence of a K+ gradient similar to that found for ATPase activity. The reaction is specific for ATP and requires cation in the sequence K+ greater than Rb+ greater than Cs+ greater than Na+ greater than Li+ and inhibited by ATPase inhibitors such as N,N'-dicylclohexyl-carbodiimide. Maximal H+ uptake occurs with an outward K+ gradient but the minimal apparent KA is found in the absence of a K+ gradient. The pH optimum for H+ uptake is between 5.8 and 6.2 which corresponds to the pH range for phosphroylation of the enzyme, but is considerably less than the pH maximum of the K+ dependent dephosphorylation. In the presence of an inward K+ gradient, protonophores such as tetrachlorsalicylanilide only partially abolish the H+ gradient but valinomycin dissipates 75% of the gradient, and nigericin abolishes the gradient. The vesicles therefore have a low K+ conductance but a measurable H+ conductance, hence a K+ gradient can produce an H+ gradient in the presence of valinomycin. The uptake and spontaneous leak of H+ are temperature sensitive with a similar transition temperature. Ultraviolet irradiation inactivates ATPase and proton transport at the same rate, approximately at twice the rate of p-nitrophenylphosphatase inactivation. It is concluded that H+ uptake by these vesicles is probably due to a dimeric (H+ + K+)-ATPase and is probably non-electrogenic.
Adenosine Triphosphatases↗
Metabolic and membrane aspects of gastric H+ transport.
Metabolic properties of dog gastric mucosa, investigated by substrate level measurements, implicate the Krebs cycle as the major energy-yielding metabolic pathway but are equivocal in terms of an ATP-based H+ secretion. Purification of gastric membranes by centrifugation and free flow electrophoresis results in a class of membrane vesicles enriched in K+-ATPase and capable of ATP-energized H+ uptake. Immunohistochemistry shows these to be derived from the parietal cell. H+ uptake by the vesicles is accompanied by K+ efflux, and movement of either ion is not potential-coupled. The simplest interpretation of these transport studies is uptake of KCl by the vesicles by passive diffusion followed by active H+:K+ exchange. In some respects, however, this model fails to conform to the expectations from in vitro studies. It may be, therefore, that another pump (i.e., redox) or another membrane component (i.e., Cl- conductance) is lost during purification. The properties of the vesicles are such, however, as to establish their role in H+ secretion by the stomach.
Adenosine Diphosphate↗
Defects of neutrophil chemotaxis and random migration in thalassemia major.
Chemotactic and random migrations in a group of 11 patients with thalassemia major were found to be defective. This may be partially the basis for the predilection to infection occasionally observed in these patients. These findings may reflect a primary defect of polymorphonuclear leukocytes or may be secondary to associated liver disease and/or diabetes mellitus. Further studies are required to define the mechanisms involved.
Adolescent↗
Asynchronous synthesis of erythrocyte membrane proteins.
The synthesis of membrane proteins of the mature mouse erythrocyte is asynchronous. During erythropoiesis, synthesis of the bulk of the spectrin and actin polypeptides is completed before that of the major transmembrane glycoprotein. Synthesis of the glycoprotein ceases before that of several minor proteins found on the inner surface of the red cell membrane, and one of these minor proteins is made predominantly by reticulocytes. These findings were the result of experiments in which a normal mouse was given a single injection of [35S]methionine. The appearance of radioactivity in the membrane proteins of circulating mature erythrocytes was followed. The earliest labeled proteins to emerge into the blood represent those synthesized at the last stages of erythropoiesis.
Animals↗
The role of nuclei and nucleoli in the control of cell proliferation.
A number of changes have been reported to occur in chromatin of quiescent cells stimulated to proliferate. These changes, indicative of increased transcriptional activity, occur in the early prereplicative phase, several hours before the onset of DNA replication, and are detectable not only in chromatin, but also in isolated nuclei and in whole cells. Most of the increased transcriptional activity can be attributed to an increased activity of the nucleolus; however, extranucleolar genes are also important in the regulation of the cell cycle flow, from G0 to S.
Animals↗
Effect on isoflurane anaesthesia and surgery on thyroid function in man.
The effect of isoflurane (Forane) anaesthesia and surgery on thyroid function was investigated in nine male patients. Isoflurane anaesthesia alone for 30 minutes prior to the start of surgery increased plasma triiodothyronine uptake (T3U) and thyroxine (T4) level by 18 and 20 per cent, respectively. Free thyroxine index (FTI) values in blood increased by 62 per cent during the same period. One hour of surgical procedure further elevated these parameters. Our data indicate that isoflurane anaesthesia and surgery increase circulating thyroxine in man.
Adult↗
Maturation of rabbit reticulocytes: degradation of specific reticulocyte proteins.
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Animals↗
Some implications of ternary diffusion in the lung.
Diffusion in the lung normally involves three gases and the governing laws are Stefan-Maxwell equations rather than the more familiar Fick's law. A simple gas film model is studied mathematically to (1) demonstrate that the rate of diffusion of a component gas may be zero even though its concentration gradient is not zero (known as "diffusion barrier"), that the rate of diffusion of a component gas may not be zero even though its concentration gradient is zero ("osmotic diffusion"), and that a component gas may diffuse against the gradient of its concentration ("reverse diffusion"); (2) compare the discrepancy between results obtained by binary and ternary laws separately; (3) determine the importance of ternary diffusion at high pressure. The findings from the model study suggest that the effects of ternary diffusion may not be pronounced when air is breathed under normal conditions, but the behavior of helium mixtures deviate significantly from that described by binary diffusion laws.
Biological Transport↗
Expression of the beta-thalassemia gene in the first trimester fetus.
To determine whether beta-thalassemia can be detected in the fetus, blood was obtained from abortuses of normal mothers and of mothers with beta-thalassemia trait. The red cells were incubated with radioactive leucine and the globin chains were analyzed by radiochromatography. Two independent methods were utilized to correct the results for contamination by maternal radioactive beta-chain, and the corrected beta/gamma ratios were compared to a previously established range of normal fetal beta/gamma synthetic ratios obtained by similar measurements in pure fetal cells. In the erythroid cells of three fetuses from mothers with beta-thalassemia trait, the beta/gamma synthetic ratio was normal in two. The third had a beta/gamma ratio of 0.04 at 10 1/2 weeks, a 50% reduction, consistent with fetal beta-thalassemia trait. Two other fetuses, derived from parents both of whom had beta-thalassemia trait, were also studied. One had a beta/gamma ratio of 0.029 at 8 weeks, a 65% reduction, also consistent with beta-thalassemia trait. The cells of the other had a ratio of essentially zero at 11 weeks, highly suggestive of homozygous beta-thalassemia. Although further experience will be needed to distinguish the homozygous and heterozygous states reliably, it now appears that the beta-thalassemia gene is expressed in the first trimester. Therefore these data suggest that the antenatal diagnosis of beta-thalassemia is becoming an attainable goal.
Female↗
Isolation and Properties of Nuclei from Control and Auxin-treated Soybean Hypocotyl.
A quick procedure for the isolation of nuclei with good yield from soybean hypocotyl (Glycine max var. Wayne) was developed. The isolated nuclei appeared to retain their structural integrity. They were typically ellipsoidal with minima and maxima diameter of about 6 and 8 to 10 micrometers. While the nuclei were similar in size, the nucleoli were significantly larger in nuclei from auxin-treated tissue. The DNA content per nucleus was 4 +/- 1 picograms for both untreated and auxin-treated tissues. The DNA: RNA: protein ratio of isolated nuclei in untreated and auxin-treated tissues was 1: 3.1: 11 and 1: 5.4: 21.7, respectively. The purified nuclei were active in RNA synthesis; the level of RNA polymerase II activity expressed in the nuclei from untreated tissue was 50 to 60% higher than RNA polymerase. I. The nuclei from auxin-treated tissues contained about 2.5 times as much RNA polymerase I activity as nuclei from untreated tissue. The purified nuclei from both untreated and auxin-treated tissues were also active in the incorporation of (3)H-TTP into DNA.
Journal Article↗