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Chemical compositions of glomerular and tubular basement membranes of human kidney.

Glomerular basement membranes (GBM) and tubular basement membranes (TBM) were prepared from human kidney and their chemical compositions were studied. Electron microscopic figures showed no contamination of the cellular components components or collagen fibers, indicating a high degree of purity of the preparations. Low contents of phospholipid supported this indication. Amino acid compositions of the human GBM and TBM resembled that of the bovine GBM, containing hydroxyproline, hydroxylsine, half-cystine, methionine and a large amount of glycine as the characteristic amino acids. The human GBM and TBM had similar carbohydrate compositions consisting of glucose and galactose, as the major sugars, together with mannose, L-fucose, hexosamine and sialic acid. Glucosylgalactosylhydroxylysine and a small amount of galactosylhydroxylysine were detected in alkaline hydrolyzates of both membranes. The human GBM and TBM separated into more than ten subunits having molecular weights ranging from 2.5 times 10-4 to more than 2.5 times 10-5 on polyacrylamide gel electrophoresis in the presence of sodium laurylsulfate and 2-mercaptoethanol. The electrophoretograms of both membranes resembled that of the bovine GBM. These observations indicate that the human TBM has a similar chemical structure to those of the human GBM and the bovine GBM.

Adult↗

Taurocholate-induced gastric damage in rats with obstructive jaundice.

We examined the severity of taurocholate-induced gastric mucosal damage in rats with obstructive jaundice. Oral administration of 100 mM taurocholate induced severer gastric mucosal damage in rats with jaundice than in intact rats. Pretreatment with 5 mM taurocholate significantly protected the gastric mucosa against 100 mM taurocholate-induced damage, suggesting adaptive cytoprotection in rats with jaundice. The reductions in the gastric mucosal potential difference and hexosamine contents in the gastric mucosa in response to 100 mM taurocholate were significantly attenuated by prior administration of 5 mM taurocholate in both intact rats and rats with jaundice.

Animals↗

Pathogenesis of water-immersion stress-induced gastric ulcers in rats with renal failure.

Rats with or without renal failure, produced by uranyl acetate, were subjected to water-immersion stress for 6 h. The severity of stress-induced gastric ulcers in the renal failure group was significantly higher than that in the control group. The intragastric pH did not significantly differ between the two groups. Transmucosal electrical potential difference, gastric mucosal blood flow, and gastric mucosal hexosamine content in the renal failure group were significantly lower than in the control group. These results suggest that decreases in defensive factors, rather than an increase in an aggressive factor, may be more closely involved in the development of acute gastric ulcers in rats with renal failure.

Acute Kidney Injury↗

Presence and chemical composition of glycoproteic layer on human spermatozoa.

Studies from electronic microscopy disclosed the presence of an electrodense stranded and branch-like electrodense layer that extends toward extracellular space. Chemical composition of this glycoproteic layer showed that protein and total sugar content is similar (0.98 microgram/microgram protein). As for the total sugar content of this glycoproteic constituent, sialic acid accounts for 40%, hexosamines 27%, and fucose 30%. Electrophoresis characterization of this constituent showed the presence of 6 different motility bands. Risen levels of sialic acid indicate the contribution of sialic residue in the net charge of the sperm membrane, its role during capacitation, and its possible participation in the formation of binding bridges between sperm membrane and ovum.

Cell Membrane↗

Content and composition of glycosaminoglycans in human knee joint cartilage. Variation with site and age in adults.

The content and composition of the matrix of human articular cartilage from two sites - the condylar summits and the upper, posterior surfaces of both lateral and medial sides - of the lower femoral epiphysis of 12 autopsy cases (age 18-46 years) were analyzed. A significantly higher content of both chondroitin sulphate and especially of keratan sulphate was found in cartilage of the condylar summits. The solubility profile of keratan sulphate showed changes indicating a difference between the metabolism of cartilage of the two sites. No significant change with side (medial or lateral) was found. A small decrease in the chondroitin sulphate to keratan sulphate ratio without change in the total amount of hexosamines with age was found on the caudal condylar summits in these adult individuals below the age of 50.

Adolescent↗

Effects of physical stress on the synthesis and degradation of cartilage matrix.

The effects of mechanical stress on the metabolism of dog femoral head cartilage have been investigated. The model involves exercising the dog on a treadmill for an 8-month period and isolating the cartilage proteoglycans from three different anatomical regions of the femoral head. These regions are 1) the articulating area, which shows degenerative changes, 2) the area surrounding the articulating area, and 3) the non-articulating area. The hexosamine analysis of cartilage from the exercised animals, indicates 43% and 21% decreases in regions 1 and 2, respectively, while in region 3 no significant change was observed. The size of the proteoglycan aggregates and monomers also decreased with exercise in regions 1 and 2, and the cartilage from these two regions also incorporated more radioactive sulfate, suggesting a possible attempt to repair the degenerative alterations. The proteoglycan monomer (D1) from the articulating region of exercised dogs failed to interact with hyaluronic acid, while the monomers from other regions in the same animal did interact. Glycosaminoglycan chains from region 1 were smaller than in the control tissue, and there was an increase in chondroitin-4-sulfate synthesis. The results of the present study indicate that the cartilage from the articulating area and the surrounding area resist mechanical deformation by increasing synthetic activity. These observations suggest that the exercised animal can be a valuable model and provide increased understanding of the mechanism of cartilage degeneration.

Animals↗

Glycosaminoglycans in localized scleroderma (morphoea).

The composition of glycosaminoglycans (GAGs) was analyzed in skin samples of eight patients suffering from localized scleroderma, i.e., three having generalized morphoea and five localized morphoea plaques. From each patients, biopsies were obtained from sclerotic and perilesional areas, and from clinically uninvolved skin of the same region. In the perilesional areas hyaluronic acid concentration was increased (p less than 0.05), while it was decreased (p less than 0.01) in sclerotic areas. Dermatan sulfate concentration was increased in the perilesional (p less than 0.05) as well as in the sclerotic (p less than 0.01) areas. Chondroitin 4/6 sulfate was increased in the sclerotic areas (p less than 0.05). Heparan sulfate showed no changes. No major differences were found in the total concentrations of uronic acid and hexosamine. This study demonstrates that changes in GAG composition in localized scleroderma follow previously described sequences of events of inflammation and fibrosis of connective tissue.

Adolescent↗

Biochemical changes caused by asbestos dust in the lungs of rats.

The contents of collagen, hexosamine, phospholipids, and cholesterol and the activities of acid and alkaline phosphatases, glutamic oxalo-acetate transaminase, glutamic pyruvate transminase, aldolase, hexokinase, and lactic dehydrogenase were determined in the lungs of rats 150 days after the intratracheal injection of amosite, anthophyllite, and chyrsotile. Anthophyllite did not cause any significant change, while amosite and chrysotile caused significant increases in the contents of collagen and mucopolysaccharides. Lactic dehydrogenase and acid phosphatase activities were increased by all the dusts, while the othe enzymes were not seriously affected. The biochemical significance of the findings in relation to abestosis was discussed.

Alkaline Phosphatase↗

[Incorporation of labeled precursors in glycosaminoglycans of the media-intima of the arterial wall].

The enzymes involved in glycosaminoglycans biosynthesis are characterized in homogenates of aortic media-intima. Incorporation of hexosamine from uridine nucleotide precursor was done into the saccharide chains of glycosaminoglycans. Labelling of aortic glycosaminoglycans by 35S-precursors is performed in our fractions; the results obtained demonstrate the uptake of 35S-sulfate from its corresponding active nucleotide sulfate ; highest sulfation is obtained for the post-microsomal supernatant. Following incorporation of 35S-sulfate or 14C-radio-activity into the different glycosaminoglycans, the largest percent uptake is obtained for heparan sulfate and the lowest for chondroitin sulfate.

Animals↗

Purification and location of the major glycoprotein of vaccinia virus.

A glycoprotein component of vaccinia virus was extracted with a non-ionic detergent NP-40 (Nonidet P-40) and purified by gel chromatography. The single antigen extracted by the detergent had a molecular weight estimated between 100,000 and 200,000 daltons. This glycoprotein was found to contain less than 1% hexosamine which would correspond to 5--10 sugar residues per molecule. Antibodies produced against this glycoprotein were able to neutralize vaccinia virus. Using immunoelectron microscopy, this molecule was found to be located in the outer layer of the virion. These results further suggest that this protein called complex E (for external) is a surface component of vaccinia virus.

Antigen-Antibody Reactions↗

[Antifibrotic properties of lidase in experimental berylliosis].

The effect of lidase with its inhalation against the background of a fibrotic process in the lungs was studied on a model of experimental berylliosis provoked in rats through intratracheal administration of 5 mg of Be (OH)2. The degree of fibrosis was assessed according to the dry residue in the lungs, content therein of oxyproline, hexosamines and hexuronic acids. Lidase was found not only to delay the development of the affections but also to lower the level of fibrosis existing prior to the treatment. The therapeutic effect of the compound continued for a comparatively long time.

Animals↗

An effect of chlorhexidine and two related drugs on glycosaminoglycans in rat skin.

The aim of the present investigation was to estimate the local effect of three plaque-inhibiting drugs on some glycosaminoglycans in rat skin. The drugs comprised chlorhexidine, dichlorhexidine, and trichlorhexidine; and the glycosaminoglycans examined were hyaluronic acid, chondroitin sulfate, and heparin. Glycosaminoglycans were measured by the carbazole method. Upon subcutaneous injections in rats, chlorhexidine, dichlorhexidine, and trichlorhexidine caused marked reductions in the surrounding skin's content of total hexosamine as well as of the three types of glycosaminoglycans. No significant difference could be demonstrated between the different types of glycosaminoglycans in this respect.

Animals↗

Methallibure inhibition of testicular and seminal vesicle activity in catfish, Clarias batrachus (Linn.): a study correlating changes in serum sex steroid profiles.

The administration of methallibure (2 microg/g BW, daily for 15 days) in Clarias batrachus in prespawning phase (May-June) resulted in decreased weights of seminal vesicle (SV) and testis, and reductions in the concentrations of total proteins, fructose, hexosamines, and sialic acid in SV and testis. The inhibitory changes can be attributed to impairment of steroidogenesis, serum levels of testosterone and estradiol -17beta decreased significantly. Withdrawal of methallibure treatment for 7 and 15 days resulted in gradual recovery and restoration of all the above parameters except the sialic acid levels in the SV and testis, and fructose level in the SV. The methallibure induced regressive changes in the SV and testis were discussed in the light of its GTH inhibiting property.

Animals↗

Effects of prednisone on gastric secretion and development of stomach lesions in swine.

In pigs treated with various doses of prednisone, pathologic changes appeared in the glandular region of the stomach, mainly in the fundic area and rarely and less severely in the cardiac and pyloric mucosae. The characteristic lesion was hemorrhagic gastritis; this was associated with massive gastric hemorrhages in pigs given the larger doses of prednisone. Epithelial changes were found in the esophageal area, but there were no ulcers. In pigs with gastric fistulas, prednisone administered at dose levels of 10 mg/kg of body weight for 8 days produced a significant increase in the volume and the acidity of gastric secretion. Hexosamine output, but not concentration, increased significantly during the treatment period. Change in pepsin secretion was sit significant. Prednisone did not produce an immediate change in gastric secretion--there being a latent period lasting 1 to 2 days before marked changes were observed. After peak changes occurred (3rd or 4th day of treatment), acidity gradually decreased to pretreatment level toward the end of the treatment period. This indicates that although increased acidity may be involved in the development of lesions in the glandular region, other factors probably have a role also in the pathogenesis of steroid-induced stomach lesions in swine.

Animals↗

Autoantigens in human sweat: purification and characterization of the D-group antigens.

We have found several heteropolysaccharides in human thermal sweat which are both autoantigenic and isoantigenic for a majority of healthy adults. They are present in trace amounts and are difficult to isolate. We have facilitated extraction through the use of a DEAE-Sephadex batch technique. One of the major antigenic fractions, the D group, has been purified and characterized. It contains 88.5 per cent carbohydrate as uronic acid, hexosamine, methyl pentose, and glucose. The protein content was 12.5 per cent.

Antigens↗

Changes in the structure of thyroglobulin following the administration of thyroid-stimulating hormone.

In each of three separate experiments, female guinea pigs in groups of 20 were given 4 units of thyroid-stimulating hormone (TSH) each day for 3 days, while controls were given saline. Na125I was injected on the 3rd day, and the animals were killed 22 hours later. The pooled throids of each group were homogenized, and thyroglobulin was purified by one of the following methods: gel filtration on Sephadex G-200 followed by density gradient ultracentrifugation, two sequential filtrations on 4 percent agarose, or filtration on 4 percent agarose followed by Sephadex G-200. TSH administration was associated with the folling changes in thyroglobulin: (1) an increase in the ratio of tri-iodothyronine to thyroxine; (2) a decrease in dissociation of the 19 S to the 12 S form; (3) an alteration in its pattern on gel electrophoresis in sodium dodecyl sulfate-urea; and (4) changes in its amino acid composition, with significant increases in the content of lysine (by 15 percent), isoleucine (by 15 percent), and methionine (by 7 percent) relative to leucine. Over-all, there were no significant changes in the content of iodine, fucose, hexosamine, or sialic acid. These data show that TSH can alter the composition of thyroglobulin independently of its effects on iodine content. We suggest that these changes may stem from alterations in the subunit composition of thyroglobulin. There were also small but significant variations in amino acid composition among the three preparations of thyroglobulin from saline-treated animals and among the three from the TSH-treated. This finding shows that thyroglobulin can be heterogeneous in its protein portion as well as in its iodine content.

Amino Acids↗

[Effect of parenteral administration of melatonin on the biochemical composition of the rat granulation tissue].

The influence of long term parenteral melatonin administration on the biochemical composition of granulation tissue of surgical wounds in rats during healing was investigated. Physiological solution and solutions containing various concentrations of melatonin were subcutaneously injected to the animals within 3 weeks. Control and injected animals were wounded. Samples of gramilation tissue were investigated on the 5-th and the 8-th day of healing. The contents of oxyprolin, uronic acids, hexosamines, total lipids and their fractions, fractional composition of glycosaminoglicans and proteins composition of salt extracts were determined in the se samples. Repeated injections during three weeks caused the changes in biochemical composition of researched samples which were characteristic for stressful reaction of connective tissue. The specific changes are most expressed at long term introduction of a physiological solution to animals. The introduction of melatonin during similar period cansed protective effect, partially defending biochemical composition of granulation tissue from changes, which were induced by stressful situation.

Animals↗

Effect of hypervitaminoisis A on composition of chick cartilage.

1. Day-old chicks were fed a diet containing one mg of retinyl acetate per kg feed. One group was treated with 18.300 mug retinyl acetate/100 g of body weight/week, whereas the other group served as control. Retinol treatment did not affect weight gain during a 4-weeks observation period. 2. At the end of this period the animals were killed and epiphyseal cartilage of the tibiotarsus examined. Retinol excess decreased ash and hydroxyproline and increased glycosaminoglycans and uronic acid in dry, fat-free cartilage. Sialic acid and hexosamine contents of glycosaminoglycans were elevated without a great change in the proportion of glucosamine and galactosamine. 3. Activity of 5 enzymes degrading glycosaminoglycans was not affected by retinol excess. 4. In hypervitaminotic chicks, uptake of injected radiosulphate by cartilage tissue and incorporation into glycosaminoglycans were enhanced and breakdown retarded.

Acetylglucosaminidase↗