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C Garbarsch

Publications and source records attributed to C Garbarsch.

At least 19 recordsLinked to original sources
PubMed1992

Quantitative PAS assay of some carbohydrate compounds and detergents.

A spectrophotometric method for determination of color development of glycocompounds subjected to PAS reaction was investigated with various carbohydrate compounds and related chemicals. The conditions of the oxidation with periodic acid was found to influence the amount of the colored Schiff dye produced. Mono- and di-saccharides (mannose, glucose and maltose) were PAS-negative. Glycogen was more reactive than dextran. When glycogen was hydrolyzed by amylase the intensity of the PAS product dropped until a certain limit probably reflecting the limit dextrin. The presence of proteins (albumin) or electrolytes (NaCl) did not influence the PAS reaction. Many non-ionic detergents commonly used in membrane biology such as alkyl glycosides and gluco-methyl alkanamides were strongly PAS-positive and so was the anionic detergent SDS while the zwitterionic detergents tested, such as CHAPS and CHAPSO, were PAS-negative. The color development of the spectrophotometric PAS reaction showed linearity with the concentration of a simple glycoprotein solution (peroxidase) and a complex solution (bovine serum). By the PAS reaction it was also possible to measure the content of soluble and membrane bound carbohydrate compounds in a pellet of liver cell membranes. We find that the PAS reaction is sensitive and reliable for quantitative estimations of complex carbohydrates as well as soluble and membrane-bound carbohydrate compounds. The latter should be treated with PAS-unreactive zwitterionic detergents.

Animals↗
PubMed1992

Changes of soluble glycoproteins in dystrophic (dy/dy) mouse muscle shown by lectin binding.

Lectin binding sites in skeletal muscle from normal and dystrophic (dy/dy) C57 BL/6J mice were demonstrated by use of histochemistry and electrophoresis combined with electron microscopy. The following lectins were used: Canavalia ensiformis Con A, Triticum vulgaris (WGA), Glycine max (SBA), Griffonia simplicifolia (GS II), Arachis hypogaea (PNA), Pisum sativum (PSA) and Lens culinaris (LCA). After incubation of frozen sections with Con A, WGA, GS II, PSA and LCA a sarcoplasmic staining was observed in both normal and dystrophic muscle. The most consistent light microscopic observations in the dystrophic muscles were a decreased staining intensity of the sarcoplasm after incubation with Con A, WGA, PSA and LCA, but not with GS II, and a strong staining of the interfiber connective tissue. Supernatants, deprived of organelles and membranes, were prepared from normal and dystrophic muscle by high speed centrifugation. Lectin stained Western blots of the supernatant from dystrophic muscle showed two bands (120 and 67 K) with high affinities to avidin. Further this supernatant contained two glycoprotein bands (180 and 140 K) with affinities to Con A and a number of glycoprotein bands with apparent molecular weights below 67 K showing affinities to LCA and PSA. None of these glycoprotein bands could be detected in the supernatant from normal muscle. These changes of the muscle carbohydrate components might be involved in the expression of the dystrophic syndrome This seems to be the first report on changes of soluble glycoproteins in muscular dystrophy.

Animals↗
PubMed1991

Lectin binding in skeletal muscle. Evaluation of alkaline phosphatase conjugated avidin staining procedures.

Cryostat sections from rat gracilis muscles were incubated with different biotinylated lectins: Con A (Concanavilin A), WGA (Wheat germ agglutinin), SBA (soybean agglutinin), GS I and GS II (Griffonia simplicifolia agglutinin), LCA (Lens culinaris agglutinin), PNA (peanut agglutinin) and PSA (Pisum sativum agglutinin). The sections were subsequently treated with alkaline phosphatase conjugated avidin. The lectin binding sites were visualized after incubation in substrate media containing: (1) 5-bromo-4-chloro indoxyl phosphate and Nitro Blue tetrazolium or copper sulphate; (2) naphthol AS-MX phosphate or naphthol AS-BI phosphate and various types of diazonium salts; (3) alpha-naphthylphosphate and Fast Blue BB; (4) beta-glycerophosphate according to the method of Gomori. The results obtained with the alkaline phosphatase methods were compared with those seen with a streptavidin-horseradish peroxidase procedure. Several chromogen protocols for visualizing alkaline phosphatase activity showed differences in the ability to detect lectin binding sites. A sarcoplasmic reaction was evident for Con A, GS II, WGA, LCA, and PSA after incubation in the indoxyl phosphate medium. Sarcoplasmic reaction for GS II was also noticed after incubation with naphthol AS-MX Fast Blue BB and beta-glycerophosphate. The latter substrate also gave rise to a sarcoplasmic Con A reaction. With the indoxylphosphate tetrazolium salt method some muscle fibres showed a very strong intracellular reaction after incubation with Con A and GS II while the staining intensity was weak in other fibres. The same muscle fibres were stained with PAS. No sarcoplasmic reactions were observed with either naphthol phosphate media or with the diaminobenzidine peroxidase methods. Further, the staining of the muscle fibre periphery, connective tissue, an capillaries was intensified using the indoxyl method. The indoxylphosphate-tetrazolium salt method seems to be suitable for future investigations of lectin binding sites in muscle sections.

Alkaline Phosphatase↗
PubMed1990

Collagen distribution in developing experimentally induced granulation tissue. A morphometric study.

Viscose cellulose sponges were implanted subcutaneously on the back of full-grown Sprague-Dawley rats. Seven, 14, 21, 28, 42, 60 and 90 days after implantation, groups of 12 animals decapitated and the sponges were removed and processed for light microscopy. Five microns sections were stained with Picro-Sirius Red. Morphometry was performed on the zone of ingrowth and the collagen. The intersectional variation in the morphometrically determined collagen density within the sponges was below 20%. The hydroxyproline content was determined biochemically in 5 microns sections of sponges implanted for 14, 42, 60 and 90 days. A positive correlation (rho = 0.79, p less than 0.0001) was observed between the biochemically and morphometrically determined collagen contents. The morphometric determinations showed a steady increase in the granulation tissue ingrowth. At day 60 the ingrowth was complete. There was an increasing collagen density from days 7 and 14 through days 21 and 28, followed by a nearly steady state up to day 90 and a significantly higher collagen density peripherally than centrally in the day 42 sponges. The study has shown that morphometric collagen determination at light microscopical level using Sirius Red-stained sections may add quantitative data describing the dynamic changes in collagen content and distribution within developing granulation tissue.

Animals↗
PubMed1989

Injury and repair of smaller muscular and elastic arteries. Immunohistochemical demonstration of fibronectin and fibrinogen/fibrin and their degradation products in rabbit femoral and common carotid arteries following a dilatation injury.

Indirect immunoperoxidase staining for fibrinogen/fibrin and fibronectin was performed on normal and healing arterial tissue of muscular and smaller elastic arteries. Fibronectin was observed in the wall of the normal arteries, whereas fibrinogen/fibrin could not be demonstrated. Fibronectin was observed in the intima as well as the media deposited in a similar fashion in the femoral and carotid artery during repair. Apart from the early occurrence of fibrin/fibrinogen in the media of both arteries the distribution of fibrinogen/fibrin and degradation products differed. In the femoral artery a progressively weakening positive reaction for fibrinogen/fibrin and degradation products towards the lumen was observed in the intima and the media 7 and 14 days after the lesion. By 28 days the reaction in the media was negative. No thrombus formation was observed. In contrast, all the specimens examined from the common carotid arteries were obliterated by luminal thrombi 28 days after the lesion. The thrombus as well as the damaged intimal thickening and the compressed media were loaded with fibrinogen/fibrin and degradation products. The deposition of fibronectin, fibrinogen, and degradation products in the carotid artery was similar to that previously reported in experimental aortic arteriosclerosis in rabbits as well as in giant cell arteritis.

Animals↗
PubMed1988

Collagen type IV and procollagen type III during granulation tissue formation: a serological, biochemical, immunohistochemical and morphometrical study on the viscose cellulose sponge rat model.

The serum concentrations of collagen type IV,7S, collagen type IV,nc1, and aminoterminal type III procollagen peptide immunoreactive components were measured by means of specific radioimmunoassays during development of granulation tissue in rats. The results were compared with tissue deposition of basement membranes and interstitial collagens in the granulation as measured morphometrically. A parallel sequential pattern in tissue deposition of collagen types III and IV, and serum increase of collagen types III- and IV-related fragments, was observed. Serum collagen type IV was less sensitive as a marker for development of granulation tissue than the serum procollagen type III N-peptide. This was in accordance with a low collagen type IV/interstitial collagen ratio in the granulation tissue. However, a cross-sectional study showed that serum collagens types IV,7S and IV,nc1 may be useful as early quantitative indicators of granulation tissue formation. Simultaneously, measurement of collagen type IV- and procollagen type III N-peptide-related antigens in serum provides a differentiated reflection of the dynamic matrix processes in developing granulation tissue.

Animals↗
PubMed1988

Serum aminoterminal type III procollagen peptide. Relation to biosynthesis of collagen type III in experimentally induced granulation tissue in rats.

Serum aminoterminal type III procollagen peptide was measured in rats during the development of granulation tissue induced by subcutaneous implantation of viscose cellulose sponges. Active collagen type III synthesis in granulation tissue during the first three weeks was accompanied by an increase in serum propeptide level. A positive correlation was observed between the increase in serum propeptide level on the one hand and the increase in granulation tissue collagen type III content and the in vitro formation of tissue 3H-hydroxyproline on the other hand. In some animals the serum propeptide level remained low, despite biochemical signs of collagen synthesis, indicating variations in the release into serum and/or the metabolism of circulating propeptide. The increase in propeptide antigen concentration was mainly due to an elevated content of material with molecular weight equal to or twice that of the propeptide. A minor fraction of the propeptide remained attached to the interstitial collagen fibres in the granulation tissue. The correlation between the serum propeptide level and the biosynthesis of collagen at the site of the focal fibroproliferative process suggests that the serum propeptide level may be a valuable indicator of fibrogenesis and thereby of disease activity in fibrotic conditions.

Animals↗
PubMed1987

Injury and repair of smaller muscular and elastic arteries. A light microscopical study on the different healing patterns of rabbit femoral and carotid arteries following dilatation injuries by a balloon catheter.

26 rabbits of the Danish country strain were subjected to mechanical dilatation injury of the left femoral and carotid arteries with Fogarty's embolectomy catheters F2 and F3 respectively. The rabbits were killed 2, 7, 14 and 28 days after the dilatation injury and the arteries examined histologically. Initially both of the arteries exhibited necrosis of the media and infiltration of the vessel wall with neutrophils and mononuclear cells. From day 7, intimal thickening was observed in both types of arteries, progressing in thickness during the later stages. However, thrombosis occurred in the majority of the carotid arteries, whereas this was only infrequently seen in the femoral arteries. In all of the dilated arteries, the elastic laminas were stretched or fragmented and never regained their normal appearance. In the carotid artery, giant cells accumulated around the fragmented elastin and calcified areas, located primarily at the intima-medial border. These changes were never observed in the femoral artery. At the twenty-eight days stage, proliferation of the smooth muscle cells more or less led to restitution of the media in the femoral artery, whereas the carotid artery showed medial restitution only to a lesser extent. The similarities between the injured carotid artery and human temporal arteritis, and the utility of the model as an animal model for the study of temporal arteritis are underlined.

Angioplasty, Balloon↗
PubMed1987

Giant-cell arteritis. Histological, immunohistochemical and electronmicroscopic studies.

Biopsies from the temporal artery of 32 patients suspected of giant-cell arteritis were evaluated retrospectively by light microscopy, histochemical, and immunohistochemical methods, as well as by transmission electron microscopy (TEM). At the clinical follow-up the 32 patients included four clinical groups: temporal arteritis (8 patients), polymyalgia rheumatica (10 patients), rheumatoid arthritis (4 patients), and a group of miscellaneous diseases unrelated to inflammatory rheumatic diseases (10 patients). There were a number of similarities between age-related alterations in the arteries and the changes in giant-cell arteritis. The most important differences were the inflammatory cellular infiltration of the media, the perifocal accumulation of fibronectin, and the occurrence of deposits of fibrin/fibrinogen and fibrin/fibrinogen degradation products. In addition, alpha-2 macroglobulin, lysozyme and factor VIII were also noted in giant-cell arteritis. The alterations in giant-cell arteritis show a number of similarities to the changes following experimental vascular injury of the rabbit aorta. The nature of the findings in human giant-cell arteritis, as well as the similarity to the experimental arteritis, indicate that giant-cell arteritis may reflect a non-specific reaction to injury, independent of the cause of the disease.

Aged↗
PubMed1986

Immune-induced vascular connective tissue alterations in rabbits chronically immunized with bovine serum albumin: biochemical studies on collagen, glycosaminoglycans, RNA, and DNA in normal and injured aorta.

Repeated intravenous injections of bovine serum albumin in rabbits caused a significant reduction in the aortic in vivo biosynthesis of chondroitin-4,6-sulfate, whereas no changes were observed in the synthesis of other glycosaminoglycans nor in the content of collagen. This contrasts the biochemical changes generally seen in acute vascular injury. When experimentally elicited vascular injury and repair processes were induced in chronically immunized rabbits, the proliferative response was greatly inhibited, as reflected by a significant diminution of the DNA amount. Also, the vascular connective tissue matrix repair was restrained: the aortic content of collagen and the collagen type III/I ratio was repressed, and the in vivo biosynthesis of sulfated glycosaminoglycans was markedly reduced. All immunized rabbits developed antibodies to bovine albumin, but in only a few were circulating immune complexes detected. The inhibitory effect of persistent immunostimulation on the nonspecific processes of repair in vascular connective tissue may be of significance as to chronicity of vasculitis, as well as inflammation and repair in general, in inflammatory connective tissue diseases.

Animals↗
PubMed1986

Immune-induced vascular connective tissue alterations in rabbits chronically immunized with bovine serum albumin: morphological and morphometric studies on normal and injured thoracic aorta.

The effect of persistent immunostimulation on normal and mechanically injured thoracic aorta was investigated histologically, histochemically and morphometrically. In the uninjured vessel wall no alterations suggestive of acute inflammation were observed following immunization, in accordance with previous biochemical studies. When mechanically elicited vascular injury and repair processes were induced in chronic immunostimulated rabbits, the neo-intimal aortic smooth muscle cell nuclear volume fraction of the vessel wall was significantly repressed, indicating, that the proliferative response to injury was inhibited. Further, the neo-intimal volume fraction of the vessel wall was reduced, suggesting impaired matrix neoformation. A highly significant linear correlation existed between the biochemically estimated DNA concentration and the nuclear volume fraction of smooth muscle cells in the vessel wall (r = 0.6275, P = 5 X 10(-5). Thus, the present study confirms previous biochemical observations, that the early processes of vascular inflammation and repair, i.e. smooth muscle cell proliferation and matrix accumulation, is inhibited following persistent immunostimulation. In addition to describing the histological correlates to the biochemical findings, important regional differences were quantified.

Animals↗
PubMed1985

Vascular injury compared to ageing of normal rabbit aorta. Biochemical and histochemical studies on time-dependent alterations of vascular connective tissue.

Male albino rabbits of the Danish country strain, 5 months of age, were divided into two groups. One group of animals was killed 180 days after a single mechanical dilatation injury of the thoracic aorta. A second group of untreated controls was killed at ages of 150, 165, 180, 210, 330, and 450 days. Glycosaminoglycans, uptake of 35S-sulphate, collagen, uptake of 125I-albumin, and vascular histochemistry and morphology were analyzed in the thoracic aorta. In the injured aortae the dry weight and the total amounts of hexosamine, hyaluronic acid, chondroitin-4,6-sulphate, dermatan sulphate, heparan sulphate, and hydroxyproline were increased. The concentration of hyaluronic acid decreased, whereas the concentration of dermatan sulphate increased. The concentrations of chondroitin-4,6-sulphate and heparan sulphate were unchanged. The total uptake of 35S-sulphate into the sulphated proteoglycans as well as the uptake of 125I-albumin were increased. The light microscopical examination showed thickening of the intima, medial changes with fibrosis, accumulations of proteoglycans, calcifications, formation of cartilage, and ossified tissue with haematopoiesis. In the uninjured thoracic aorta the only significant change during ageing was an increase in the total amount of hyaluronic acid and a decrease of the 35S-sulphate incorporation into the chondroitin-4,6-sulphate in the aorta. No morphological or histochemical alterations were observed during ageing. Spontaneous lesions were observed in 2 out of 55 aortas. It may be concluded that injury and ageing are reflected quite differently in the thoracic aorta of the rabbits. The observations may be of relevance to the interpretation of the alterations in human arterial diseases involving processes of injury and repair as well as ageing.

Aging↗
PubMed1979

Increased collagen prolyl hydroxylase activity in the aortic wall of rabbits exposed to chronic hypoxia.

The activity of collagen prolyl hydroxylase in aortic wall was studied in rabbits exposed to chronic 10% ambient oxygen tension for 30 days. Prolyl hydroxylase in rabbit aorta was shown to be similar to the enzyme from other sources in that it required molecular oxygen, alpha-ketoglutarate, ferrous iron and ascorbate for its activity. The activity of prolyl hydroxylase was increased to 180% of controls in the intima-media samples from rabbits exposed to hypoxia. No atherosclerotic lesions could be seen in arteries of animals kept in chronic hypoxia. If the arteries of rabbits were injured with a single mechanical dilatation, the activity of prolyl hydroxylase increased more than 2-fold, as reported previously. The exposure of these animals to chronic hypoxia further elevated the prolyl hydroxylase activity.

Animals↗
PubMed1979

Endothelial injury of arteries following catheterisation with polyethylene tubes: experimental studies on rabbit aorta using the Seldinger technique.

The effect of catheterisation on the rabbit aortic wall is investigated employing a technique identical to that used for continuous monitoring of blood pressure in humans. The catheters were introduced into the aorta through the right femoral artery of the rabbit using the Seldinger technique, except that the catheters were introduced directly into the femoral artery and not percutaneously. It was found that catheterisation for 24 h was followed by intimal and medial injuries. 3 to 60% (mean 20.4%) de-endothelialisation of the aortic surfaces was observed. The present experiments appear to explain some of the complications observed after continuous registration of blood pressure in humans. Case histories are referred to in which the brachial artery was catheterised.

Animals↗
PubMed1979

Formation of granulation tissue in subcutaneously implanted sponges in rats. A comparison between granulation tissue developed in viscose cellulose sponges (Visella) and in polyvinyl alcohol sponges (Ivalon).

A comparison was made between the granulation tissue formation in two different synthetic sponge types. Visella and Ivalon, of different sizes. The granulation tissue formed in the two sponge types did not differ qualitatively, and had the character of wound tissue and inflammatory tissue in man. The rate of tissue formation in the Visella sponges was faster and the tissue was more homogenous than in the Ivalon sponges. Fourteen-day-old Visella implants of either size contained more granulation tissue than Ivalon sponges, probably owing to the smaller pore size of the former material. This may also account for the more frequent occurrence of giant cells in the Visella implants. In contrast to the Visella sponges, the trabeculae of the Ivalon polymer showed calcification and positive staining properties with histological staining procedures, and deformation was frequent among the Ivalon implants. Thin sponges of either type closed in about 21 days, thick ones after about 42 days of implantation. Calculated per 2 cm3 of implant, thin sponges produced more tissue after 14 days of implantation than thick noes. It is concluded that the Visella sponge type is best suitable for this experimental model of inflammation.

Animals↗
PubMed1978

Succinate dehydrogenase activity in the wall of rabbit aorta. The histochemical use of PMS and exogenous coenzyme Q10 as intermediate carriers.

An investigation of succinate dehydrogenase activity in the wall of rabbit aorta was carried out. The level of succinate dehydrogenase per se in the smooth muscle cells was found to be fairly high, while the mitochondrial level of carrier CoQ was low. The latter may explain the low level or lack of activity of succinate dehydrogenase in these cells as noticed by previous authors. A reliable image of the actual level of succinate dehydrogenase was obtained only by adding CoQ10 to the incubation system. PMS should be avoided, as it induced a "Nothing dehydrogenase" reaction even at low concentrations.

Animals↗