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Biomedical subjects

A Chung

Publications and source records attributed to A Chung.

At least 73 records · Page 4Linked to original sources

Fine structure of the glomerular basement membrane and immunolocalization of five basement membrane components to the lamina densa (basal lamina) and its extensions in both glomeruli and tubules of the rat kidney.

Electron microscopic immunostaining was used to examine the localization of type IV collagen, laminin, entactin , heparan sulfate proteoglycan, and fibronectin within the basement membranes of the rat kidney. In preliminary experiments, various methods of processing formaldehyde-fixed kidney were compared using antilaminin antiserum and the indirect immunoperoxidase method. Little or no laminin immunostaining of the glomerular basement membrane was present in sections unless they had been frozen-thawed; and even in this case, the immunostaining was light in comparison to that of basement membranes in adjacent tubules. However, when frozen-thawed sections were treated with 0.5% sodium borohydride, immunostaining was then as strong in glomerular as in tubular basement membranes. Accordingly, this treatment was applied to frozen-thawed sections before immunostaining for any of the substances under study. Immunostaining of the glomerular basement membrane for each of the five substances was fairly uniform throughout the lamina densa (also called basal lamina), but uneven in the lamina lucida interna and externa (also called lamina rara interna and externa) in which stained bands extended from the lamina densa. Similarly in the basement membranes of tubules, immunostaining for the five substances was localized to the lamina densa and bands extending into the lamina lucida. When the ultrastructure of the glomerular basement membrane was examined, three structures were found: (1) a network of 4-nm-thick "cords," which seems to be the main component; the cords are closely packed in the lamina densa and more loosely arranged in the lamina lucida interna and externa; (2) straight, hollow 7-10-nm-thick structures referred to as " basotubules "; and (3) 3.5-nm elements composed of minute paired rods, referred to as "double pegs." The distribution of the cords, but not that of the other two structures, was related to the immunostaining pattern. It is concluded that (1) to fully reveal the antigenicity of the glomerular basement membrane, frozen-thawed sections must be treated with sodium borohydride prior to immunostaining, possibly because this basement membrane is more compact than the others; and (2) in both glomerular and tubular basement membranes, type IV collagen, laminin, entactin , heparan sulfate proteoglycan and fibronectin are colocalized in the lamina densa and its extensions to the laminae lucidae . Since the distribution of the cords corresponds to that of immunostaining, it is likely that the five substances are present within the cords.

Animals↗

A third example of haemolytic auto-anti-Vel.

A non-transfused, 43 year old Caucasian female presented with acute haemolytic anaemia and splenomegaly. Sections of bone marrow showed erythroid hyperplasia. The patient's red blood cells gave a negative reaction with polyspecific antiglobulin serum, but a positive reaction with specific anti-IgM. A heat eluate prepared from her red cells showed anti-Vel specificity. Her serum agglutinated only Vel-positive cells including her own. All papain pre-treated red cells including her own and Vel-negative cells were completely haemolysed at 37 degrees C. The percentage of haemolysis of Vel-positive cells was greater than that of Vel-negative cells.

Adult↗

Assessment of methods for cytomegalovirus antibody testing of blood donations.

As part of a programme to establish a panel of individuals with high titres of anticytomegalovirus (CMV) IgG for the production of CMV Ig, the CMV serostatus of 139 normal blood donors was determined by indirect haemagglutination assay, enzyme immunoassay, and complement fixation. The indirect haemagglutination assay was the most accurate and convenient method for the detection of CMV antibody negative donations. Plasma for production of hyperimmune CMV IgG could best be selected by an IgG-specific enzyme immunoassay procedure.

Antibodies↗

The upper and lower bounds of rate constants for general mammillary compartment systems.

The upper and lower bounds of rate constants for general mammillary three and four compartment systems have been derived. It is further proposed that the midpoints of the bounds can be used as initial estimates for parameter estimation. Numerical examples are given demonstrating the closeness of the calculated midpoints to the "known" rate constants of both the three and four compartment systems.

Kinetics↗

Detection of anti-varicella-zoster virus antibodies in blood donors by automated passive haemagglutination.

1,219 normal blood donors were screened for anti-varicella-zoster virus antibodies by automated passive haemagglutination (APH) on a Technicon 15-channel AutoAnalyzer. 98 samples (8%) were positive by APH and 92% of these were shown to have antibody titres greater than or equal to 1/16 by indirect immunofluorescence assay (IFA). Parallel testing on an additional 265 donors revealed that 11 (4%) individuals had IFA titres greater than 1/16 and 6 of these 11 donors were identified by APH. Thus, APH is an effective means of rapidly detecting units of plasma acceptable for production of varicella-zoster immunoglobulin.

Antibodies, Viral↗

Angiotensin-converting enzyme: I. New strategies for assay.

The disposition of converting enzyme (kininase II) on the luminal surface of pulmonary endothelial cells is well established. Further, it is known that there is a net conversion of angiotensin I into angiotensin II as blood passes through the lungs. However, little is known about modulations of converting enzyme activity that may arise through, e.g., changes in the quality of inhalants, blood flow, or blood oxygenation. There are few data on the effects of lung disease. A major barrier to studies to examine for pathophysiologic modulations of converting enzyme is that of assay. The enzyme can be measured in terms of the rate of formation of angiotensin II from a known quantity of angiotensin I. However, both peptides are biologically active, and lungs contain other enzymes capable of degrading them. We have developed a series of radiolabeled, acylated tripeptides to improve our ability to examine for changes in the net converting enzyme of intact lungs. The enzyme, a dipeptidyl carboxypeptidase, is capable of removing C-terminal dipeptides from a variety of oligopeptides. We have prepared benzoyl-Gly-Gly-Gly (I), benzoyl-Pro-Phe-Arg (II), benzoyl-Gly-His-Leu (III), benzoyl-Phe-Ala-Pro (IV), and benzoyl-Phe-His-Leu (V), each containing a (3)H-atom in the para position of the benzoyl moiety. Substrates I and III have been used previously in photometric assays of low sensitivity. II is the acylated C-terminal tripeptide of bradykinin, IV is an acylated tripeptide analog of BPP(5a) (<Glu-Lys-Trp-Ala-Pro) and V is the acylated C-terminal tripeptide of angiotensin I. These substrates can be used in vitro or in vivo to measure converting enzyme. The (3)H-labeled product is separable by partitioning between an organic solvent and acidified aqueous solution. The product is quantified by scintillation counting of the organic phase. The choice of substrate depends on the goals of the experiment: substrate I or III when wide variations in substrate concentrations are needed but high sensitivity is not; substrate IV when high sensitivity is needed.

Angiotensin I↗

Biliary lipid synthesis and secretion in gallstone patients before and during treatment with chenodeoxycholic acid.

The interrelationships between biliary lipid secretion and the hepatic activities of the rate-limiting enzymes for bile acid and cholesterol synthesis have not been investigated in patients with gallstones before and during desaturation therapy. Liver biopsies for enzyme assays and biliary lipid secretion measurements were performed in 12 patients with gallstones before chenodeoxycholic acid therapy and in nine of these patients at 9 months of therapy. Six nongallstone control patients underwent only the lipid secretion measurements. In the patients with gallstones before treatment, all of whom had saturated bile, increased cholesterol secretion correlated directly with increased HMGCoAR activity, whereas bile acid and phospholipid secretion rates were significantly lower than in controls. During desaturation in response to chenodeoxycholic acid, biliary cholesterol and phospholipid secretion rates decreased significantly, and bile acid secretion was unchanged. Concomitantly, both HMGCoAR and cholesterol 7alpha-hydroxylase activities decreased significantly, but the correlation between HMGCoAR and cholesterol secretion was lost. Furthermore, no correlation was found between cholesterol 7alpha-hydroxylase activity and bile acid secretion during therapy. Enzyme assays were performed on single liver samples obtained at the same time of day but 48 hr before the lipid secretion measurements. That correlations between data obtained under these conditions are valid remains to be proved. In conclusion, the mechanism of biliary cholesterol saturation in patients with gallstones probably is related primarily to increased hepatic cholesterol synthesis, whereas desaturation during chenodeoxycholic acid therapy involves altered relationships among hepatic enzyme activities and biliary lipid secretions.

Bile Acids and Salts↗