Differenzierung der mucopolysaccharide des innenohres durch säulenchromatographische hexosamintrennung

Zusammemfassung Die sauren Hydrolysate der cochleären Abschnitte des häutigen Labyrinths von Hausschweinen und Meerschweinchen wurden säulen-chromatographisch zur Differenzierung des Glucos- und Galaktosaminanteils getrennt. In beiden Tierspecies ergab sich ein Überwiegen des Glucosamins gegenüber dem Galaktosamin. Unter Berücksichtigung histochemischer und quantitativer biochemischer Befunde lassen die Analysen den Schluß zu, daß stark saure MPS am Aufbau des häutigen Hörorgans in erheblichem Maße beteiligt sind. Neben dem Keratosulfat als Hauptkomponente sind auch Chondroitinsulfate vertreten. Die stark sauren Eigenschaften dieser Mucopolysaccharide unterstreicht die Theorie ihrer wesentlichen Rolle als Ionenaustauscher bei der Bildung der Innenohrpotentiale.

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Differentiation of mucopolysaccharide in the inner ear by separation of hexosamine with column-chromatography

Summary The glucose- and galactose-amino components were separated by columnchromatographic differentiation from the hydrolyzed cochlear sections of the membranous labyrinth of the pig and the guinea-pig. They showed in both species more glucosamine than galactosamine. Taken consideration of the histochemical and biochemical findings these analysis may be concluded that a considerable amount of the strong acid mucopolysaccharide is contributed in construction of the membranous hearing organ. In addition to keratosulphate as main components, the chrondroitinsulphate was also present. The strong acidic qualities of the mucopolysaccharide underlined the theory of their important role as ionic exchange upon the formation of the inner ear potential.

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Herrn Prof. Dr. Dr. h. c. A. Eckert-Möbius zum 80. Geburtstag gewidmet.

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Technische Mitarbeit: Frau W. Lehmann.     

Das LDH-lsoenzymmuster der menschlichen tonsillen und seine bedeutung für den energies toffwechsel

Zusammenfassung Das LDH-Isoenzymmuster von 40 menschlichen Tonsillen wurde elektrophoretiseh untersucht. Im Durchschnitt ergibt sich bei den Tonsillen eine etwa gleichstarke Beteiligung des aeroben wie anaeroben Energiestoffwechsels. Eine geschlechtliche Differenzierung war nicht zu erkennen. Mit zunehniendem Alter tritt im paratonsillaren Gewebe and im Kryptenepithel eine leichte Verschiebung zugunsten des anaeroben Stoffwechsels ein In Fallen chronisch rezidivierender Tonsillitis mit Fokalerkrankung war der aerobe Charakter des Energiestoffwechsels besonders im paratonsillaren Gewebe deutlich erhöht, bei Paratonsillitiden deutlich vermindert, ebenso im Kryptenepithel letzterer. Wesentliche Abweichungen von der Norm wurden bei einem Patienten mit einem Morbus Langdon-Down gefunden.

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The LDH-isoenzymes distribution in human tonsils and its consequence for the energy-metabolism

Summary The distribution of the LDH isoenzymes of 40 human tonsils has been investigated electrophoretically. The aerobic and anaerobic metabolism was observed without any sex predominance. With the advance of age the paratonsil tissue and the epithelial crypts showed a small displacement towards the anaerobic metabolism. In cases of chronic relapsing tonsillitis with vocal disease the aerobic character especially in the paratonsil tissue was distinctly increased, and in paratonsillitiden and their epithelial crypts were distinctly decreased. Abnormal results were obtained in one patient who suffered from Langdon-Down's disease.

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Technische MitarbeitE. Del Mestre u.W. Lehmann.     

Platelet Functions in Cardiopulmonary Bypass Surgery

Background

Haemorrhage after Cardio Pulmonary Bypass (CPB) Surgery is a well recognised complication that leads to significant morbidity and mortality. The incidence varies between 5-25% depending upon the clinical situation. Several factors are implicated as causative but none have been precisely proved.

Methods

Our study was an attempt to evaluate the haemostatic defect with particular reference to platelet function abnormalities during cardio pulmonary bypass surgery, in order to reduce the morbidity and mortality associated with post CPB haemorrhage. Flow cytometric evaluation of different platelet glycoproteins like GPIb/IX, GPIIb/IIIa and GMP-140 was done.

Results

The marker expression showed deregulation during surgery which returned to base after bypass was terminated. In contrast, the cases with bleeding showed significant variation. P-Selectin (GMP 140) expression decreased progressively till 3^rd post-operative day showing lack of activation of platelets in cases of severe bleeding.

Conclusion

Longer duration of CPB initiates plasmin generation through heparin, which raises the PAI-1-tPA complex and thereby down regulating the functions of platelets. This suggests a link between duration of CPB, bleeding, platelet dysfunction and fibrinolysis. Hence serial estimations of the levels of GMP-140 and tPA can predict severe bleeding.Key Words: CardioPulmonary Bypass, Platelet dysfunction, flowcytometry, platelet glycoproteins, haemorrhage     

Human articular chondrocytes exhibit sexual dimorphism in their responses to 17beta-estradiol

OBJECTIVE: The higher incidence of osteoarthritis in females suggests that there may be intrinsic sex-specific differences in human articular chondrocytes. 17beta-Estradiol (E2) regulates rat growth plate chondrocytes through traditional nuclear receptor mechanisms, but only female cells exhibit rapid membrane-associated effects mediated through protein kinase C (PKC) alpha. Here we demonstrate sexual dimorphism in the physiological response of human articular chondrocytes to E2. METHODS: Articular chondrocytes were obtained at the time of autopsy from three male and three female donors between 16 and 39 years of age. Second passage cultures were treated with E2 for 24 h to assess the effects of the hormone on [3H]-thymidine incorporation, [35S]-sulfate incorporation, and alkaline phosphatase specific activity. In addition, the chondrocytes were treated for 3, 9, 90 or 270 min and PKC specific activity was determined. RESULTS: All chondrocytes were positive for aggrecan and estrogen receptor alpha mRNAs but were negative for type II collagen mRNA. Only cells from female donors responded to E2. DNA synthesis, sulfate incorporation and alkaline phosphatase activity were increased. E2 caused a rapid increase in PKC activity in the female cells within 9 min that was maximal at 90 min. Treatment with the PKC inhibitor chelerythrine blocked these effects. CONCLUSIONS: These results provide the first definitive evidence that normal human cells exhibit an intrinsic sex-specific response to E2 and suggest that sexual dimorphism may be an important variable in assessing the pathways that modulate cell behavior.     

The effect of rapamycin on the healing of the ureteric anastomosis and wound healing

Rapamycin is a potent immunosuppressive agent that also inhibits fibroblastic activity and therefore may affect the healing of various tissues. The aim of this study was to investigate the effect of rapamycin on wound healing and the healing of the ureteric anastomosis. Large White/Landrace pigs were subjected to a laparotomy and division and immediate anastomosis of the ureter. The animals were randomly allocated to receive either rapamycin or placebo. The animals were sacrificed on postoperative day 5, and strips of the skin and fascia closure and the ureteric anastomosis excised and used to determine the tensile strength, hydroxyproline levels, and histological changes. The tensile strength and the hydroxyproline levels in the ureter and fascia were lower in the rapamycin-treated animals. There was no difference in the tensile strength in the skin, although the hydroxyproline levels were lower. This study shows that healing of the ureteric anastomosis and fascia and skin closure may be impaired by rapamycin.     

Diffraction enhanced imaging of articular cartilage and comparison with micro-computed tomography of the underlying bone structure

The goal of this study was to explore the role of diffraction enhanced X-ray imaging (DEI) for assessing changes in osteoarthritic cartilage and correlating the findings with concurrent changes in the underlying bone imaged using micro-computed tomography (CT). DEI was used to image femoral head specimens at various beam energies. DEI utilizes a monochromatic, highly collimated beam, with an analyzer crystal that selectively weights out photons according to the angle they have been deviated with respect to the original direction. This provides images of very high contrast, with the rejection of X-ray scatter. The underlying bone was imaged using CT and measures quantifying the bone structure were derived. Confirmation of cartilage degeneration was obtained from histology and polarized light microscopy. DEI allowed the visualization of articular cartilage and reflected the fibrillations and fissures in tissues from degenerated joints. The trabecular bone underlying the most degenerated articular cartilage showed increased bone volume fraction and more plate-like characteristics, compared with that underlying normal appearing cartilage. The histology and polarized light microscopy images reflected the DEI based features of cartilage architecture. These data reflect the ability of X-ray based emerging technologies to depict cartilage–bone interactions in joint degeneration.     

The Relationship of a <Emphasis Type="Italic">Helicobacter heilmannii</Emphasis> Infection to the Mucosal Changes in Abattoir and Laboratory Pig Stomach

Purpose A pig ulcer model in which ulceration is reproducibly induced in the pars oesophagea (a tongue of the oesophageal squamous epithelium that extends into the pig stomach) by bile duct ligation (BDL) was used in this study to determine whether Helicobacter heilmannii (Hh) is a predisposing factor in the ulceration of this region. The infection with Hh and its relationship to ulceration and mucus integrity was examined.Methods We microscopically investigated the occurrence of spontaneous pars oesophageal ulceration in 33 pigs from a local abattoir and 5 pigs nurtured in pens in our surgical laboratory (JSM). Further groups of 5 and 6 JSM pigs underwent a sham operation and a BDL, respectively. Giemsa staining was used to detect Hh and purified mucin was characterized by gel filtration.Results Ten of 33 and 2 of 5 of the stomachs of abattoir and JSM pigs, respectively, were positive for Hh by Giemsa stain. Three of the 33 abattoir pigs showed ulceration in the pars oesophagea and none of these was Hh-positive. All six of the bile duct-ligated pigs showed ulceration in the pars but only 2 of these were Giemsa-positive. Only 8 of 33 of the abattoir pigs had 50% large polymeric mucin that was eluted in the void/excluded volume of a Sepharose 2B column.Conclusions There was no consistent correlation between an infection of the pig stomachs by Hh, an ulceration of the pars oesophagea, and mucin degradation. There was a significant difference between the percentage of polymeric mucin from the abattoir pigs and that of the JSM group (P < 0.003), the JSM group vs sham-operated pigs (P < 0.011), and JSM vs BDL pigs (P < 0.0005), but there appeared to be no association between the infectivity with Hh and mucin degradation.