The incorporation of [ 35 S]sulfate into total and specific types of serum glyeosaminoglycans was studied in rats with acute, subacute or chronic liver injury (liver cirrhosis), and compared with that of normal rats. The macromolecular (protein-bound) nature of serum glyeosaminoglycans in normal and diseased animals was also analysed. The results show a strong increase in rate and extent of [ 35 S]sulfate incorporation into total serum glyeosaminoglycans for acutely but a decrease for subacutely and chronically liver damaged rats. The time-course of distribution of label between serum chondroitin sulfate and dermatan sulfate exhibits significant changes in liver-injured animals, in particular a relatively high proportion of dermatan [ 35 S]sulfate in rats with cirrhotic livers.In comparison with serum glyeosaminoglycans the labeling profile of glyeosaminoglycans in the cirrhotic liver was quite different (heparan sulfateidermatan sulfate:chondroitin sulfate = 1:0.34:0.09) and changed only insignificantly during a l h labeling period.The protein-bound moiety of serum glyeosaminoglycans was not affected by liver disease; but the elution profile of chondroitin [ 35 S]sulfate from Dowex 1 X 2 for treated rats was altered, thus indicating a structural modification of its carbohydrate chain. Metabolische und strukturelle Untersuchungen der Glykosaminoglykane im Serum, im Vergleich zur Leber, bei normalen und leber-geschädigten RattenZusammenfassung: Der Einbau von [ 35 S]Su,lfat in die gesamten und spezifischen Typen der Glykosaminoglykane im Serum von Ratten mit akuter, subakuter oder chronischer (Lebercirrhose) Leberschädigung wurde untersucht und mit dem gesunder Tiere verglichen. Zusätzlich würden Untersuchungen zur makromolekularen, insbesondere zur proteingebundenen Struktur der Glykosaminoglykane im Serum normaler und lebergeschädigter Ratten durchgeführt. Die Ergebnisse zeigen einen starken Anstieg der Rate und des Ausmaßes der Sulfatinkorporation in die gesamten Glykosaminoglykane im Serum bei akut, eine Erniedrigurig jedoch bei subakut und chronisch lebergeschädigten Ratten. Der zeitliche Verlauf der Verteilung der [ 35 S]Märkierung zwischen Chondroitinsulfat und Dermatansulfat im Seruni ließ signifikante Veränderungen bei lebergeschädigten Tieren erkennen, insbesondere einen relativ hohen Anteil von Dermatan [ 35 S]sülfat bei Ratten mit Leberckrhose. Im Vergleich zu Glykosaminoglykanen im Serum war das Markierungsprofil der Glykosaminoglykane in cirrhotischer Leber sehr verschieden (HeparänMfat:Dermatansulfat:Chondroitinsulfat = 1:0.34:0.09) und veränderte sich während einer l-stündigen Markierungsperiode nur geringfügig. Die proteingebundene Struktur der Glykosaminoglykane im Serum wurde durch experimentelle Lebererkrankungen nicht beeinflußt, jedoch weist die Änderung des Elutionsprofils von Chondroitin[ 35 S]sulfat an Dowex l X 2 auf strukturelle Veränderungen seiner Kohlenhydratkette bei Leberschädigung hin.
Summary: Administration of a single dose of jD-galactosamine to rats causes time-dependent, biphasic changes of total glycosaminoglycan synthesis in liver. A rapidly occurring inhibition is followed by a significantly enhanced 0> 2 fold) production of 35 S-labeled glycosaminoglycans in later stages of injury. Degree and duration of the inhibitory phase are dose-dependent; 50% inhibition is reached at 80 mg/kg and maximum inhibition (nearly 80%) at about 300 mg/kg body weight 2 h after injection of Z)-galactosamine.The hepatötoxin impairs preferentially the production of heparan sulfate, whereas that of chondroitin sulfate and dermatan sulfate is diminished only slightly and for a rather short period of time. The synthesis of the latter, however, is more stimulated than that of heparan sulfate in later stages of injury.The specific radioactivity of 35 S-labeled 3'-phosphoadenosine-5'-phosphosulfate (PAPS) did not change significantly during the course of acute liver damage.Glycosaminoglycan synthesis in regenerating liver was nearly unaffected by D-galactosamine. Uridine at the dose applied partially reversed Agdactosamine-inhibited synthesis of proteoheparan sulfate.In accordance with the labeling studies the content of glucosamine^containing glycosaminoglycans in treated liver decreased, whereas that of galactosamine-cöntainiiig glycosaminoglycans slightly increased, resulting in a nearly 50% reduction of the glucosamine/galactosamine ratio 5 h after administration of Z)-galactosamine. Ion exchange Chromatographie studies of 35 S4abeled specific types of glycosaminoglycans from normal and galactosamine-injured liver revealed only minor structural differences. Synthese der Leberglykosaminoglykane in den Frühstadien der Galaktosamin-Hepatitis: Einer raschen Erniedrigung des Heparansulfates folgt eine Erhöhung von Chondroitinsulfat und DermatansulfatZusammenfassung: Eine Einzeldpsis von D-Galaktösamin erzeugt in der Rattenleber zeitabhängige, biphasische Veränderungen der Glykosaminoglykansynthese. Eine sofort eintretende Hemmung wird gefolgt von einer signifikant erhöhten (über 2fäch) Produktion 35 S-marJaerter Glykosaminoglykane in späteren Stadien der Schädigung. Grad und Dauer der inhibitorischen Phase sind dosisabhängig; 50% der Hemmung tritt bei 80 mg/kg und maximale Hemmung (nahezu 80%) bei etwa 300 mg/kg Körpergewicht 2 Stunden nach Verabreichung von £-Galaktosamin ein. Das hepatotöxische Z^Gdaktosamin beeinträchtigt vorzugsweise die Produktion von Heparansulfat, wohingegen die Synthese von Chondroitinsulfat und Dermatansulfat nur geringgradig und für einen kurzen Zeitraum vermindert ist. Die Bildung der letztgenannten Glykosaminoglykane wird jedoch in späteren Stadien der Schädigung stärker als die von Heparansulfat stimuliert.Die spezifische Radioaktivität von 35
All heart transplant patients in our clinic received intravenous immunoglobulins as a prophylaxis against cytomegalovirus infections or reactivations. Serum was sampled from 160 heart transplant patients within 4 months after surgery. In 98 samples (61%) hepatitis C virus (HCV)-specific antibodies could be detected by a "second generation" enzyme immunoassay. Of these HCV antibody-positive patients 89 were tested for a second time. At this time, 5-11 months later, in 66 patients (74%) the HCV antibody had disappeared. In the 23 still positively reacting patients, immunoglobulins were given in the last 6 months before serum sampling. Nine commercial immunoglobulin preparations were tested for HCV-specific antibodies and the presence of HCV RNA. Seven preparations were anti-HCV positive with titres in the range of 64-256, whereas reverse transcription and polymerase chain reaction did not detect HCV RNA in any immunoglobulin preparation. Passive antibody transfer rather than a HCV infection is the cause of HCV antibody detection in our patients. The presence of HCV antibodies in high concentrations in commercial immunoglobulin preparations may only be explained by an extremely high proportion of anti-HCV-positive single donations in the plasma pools used for immunoglobulin production. The passive HCV antibody transmission prevents anti-HCV serological monitoring of patients treated with these preparations. Additionally, there are reports on the transmission of hepatitis non-A, non-B via immunoglobulin preparations. Therefore, we recommend an anti-HCV screening of plasma donors.
Objective: Determination of lymphocyte subset alterations during the early phase after heart transplantation. Comparison of lymphocyte activation parameters and concentrations of soluble class I MHC molecules with the established endomyocardial biopsy rejection grading results. Patients and Methods: 154 heart transplant patients with 382 endomyocardial biopsies during the first 3 months after transplantation. The following statistical data were calculated: sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic efficiency (sum of true-positive and true-negative results as a part of total results). The immunophenotyping by flow cytometry includes CD4+, CD8+, CD3/HLA-DR+, CD8/HLA-DR+ and CD4/CD25+ subsets. The activation index (activated lymphocytes defined by microscopy after staining) was also determined. Results: By shifting the cut-off of lymphocyte subsets to higher cell concentrations, the positive predictive values increased, reaching 60–100% for some parameters. However, the overall diagnostic efficiency was considerably lower: activation index 54%, CD8/HLA-DR+ 58%, CD3/HLA-R+ 57%, CD4/CD25+ 49%, and CD4/CD8-ratio 46%. The determination of soluble class I MHC molecules in plasma samples drawn during rejection episodes and in rejection-free periods in a group of 53 heart transplant patients showed no differences between the periods. Conclusions: Immunophenotyping by flow cytometry or cytology for detection of activated T lymphocytes in peripheral blood has a low overall diagnostic efficiency in rejection diagnosis. The determination of soluble class I MHC molecules seems to be of no value for the early detection of rejection.
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