Biomedical subjects
Publications and source records attributed to M Klinger.
An overview of literature data concerning the interactions between hypertension and vascular endothelium is presented. Arterial hypertension is accompanied by morphological and functional changes of endothelium. The laboratory methods measuring endothelial damage are very useful tools in evaluation of endothelium state.
Various hemostatic abnormalities that can be met in the course of apparently unrelated diseases are caused by antiphospholipid antibodies (APA). Nearly on half of the population with antibodies detectable in serum suffer from systemic lupus erythematosus (SLE) or the disease will be diagnosed in the future. The supposition considering that APA do not bind phospholipids directly becomes popular recently. They bind serum beta 2-Glycoprotein. The APA-beta 2 GIP complexes speed up prothrombin activation and make beta 2 GIP less available for serum C- and S-protein transformation. APA are a heterogenous population of antibodies. Postinfectious APA differ from those found in autoimmune diseases. Thrombotic events caused by APA are treated according to general principles. Steroid therapy is essential in treatment programme of SLE with APA. In severe cases it is supplemented by intravenous cyclophosphamide. Plasmapheresis and intravenous immunoglobulins are of limited usefulness. In severe resistant thrombocytopenia one can try to introduce Danazol carefully.
A novel family of structurally and functionally related polypeptides has recently been detected that are now referred to as chemokines. Within this family, a peptide with the acronym RANTES was shown to be chemotactic for memory T cells, monocytes, and eosinophilic and basophilic granulocytes, thus suggesting it plays an important role in chronic inflammatory and allergic diseases. Murine monoclonal antibodies as well as cDNA probes specific for human RANTES were raised and extensively characterized. With these antibodies, stimulated human dermal fibroblasts were shown to express intracellular RANTES peptide by immunocytochemistry. Furthermore, similar kinetics could be demonstrated in fibroblasts for both RANTES mRNA expression and secretion of RANTES peptide using Northern blot hybridization and sandwich-enzyme-linked immunosorbent assay, respectively. RANTES expression was induced upon stimulation with tumor necrosis factor-alpha as well as with interleukin-1 alpha and -beta in a concentration- and time-dependent manner. These results reinforce the role of both resident and circulating cells in the production and release of RANTES and their participation in inflammatory processes.
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According to its morphological appearance, Kikuchi's disease is also called "histiocytic necrotizing lymphadenitis". This disorder was first described in Japan in 1972 as a benign lymphadenopathy of the neck. In Germany only a few cases have been reported by pathologists, whereas most clinicians are unaware of the existence of this rare disease, which can easily be mistaken for malignant lymphoma. In 1993 three cases of Kikuchi's lymphadenitis were treated at the ENT Department of the University of Lübeck. Patients' ages ranged from 15 to 30 years old. Except for cervical adenopathy, findings in blood tests, viral serology and radiological imaging were unremarkable. While fine needle aspirations were suspicious for malignant lymphoma, a final diagnosis of self-limiting "Kikuchi's lymphadenitis" was established histologically from excised lymph nodes. In our opinion early extirpation of lymph nodes and consultation of a pathologist are necessary to make a correct diagnosis without delay. Thus, in patients with Kikuchi's lymphadenitis unnecessary staging procedures and treatment can be avoided.
The subject of analysis were data concerning the 101 pregnancies in 43 patients with systemic lupus erythematodes observed in years 1965-1990. The evaluation proved that pregnancy does not deteriorate the course of lupus nephritis that is in the remission before the conception. In lupus patients the risk of fetal loss and stillbirth is high, especially in the presence of anticardiolipin antibodies. The course of active lupus occurring during pregnancy is frequently severe and its effect on the pregnancy is very harmful.
Up to day the pathogenesis of immune deficiency in chronic renal failure have not been clearly elucidated. There is no agreement where is situated the primary disturbance that makes that category of patients more susceptible to infections and neoplasms. The views presented in this paper seem to shift the balance toward multifactor theory. Possible influence of many biochemical abnormalities and effect of renal replacement therapies on main elements of immune system are discussed.
Two hydrophilic acrylic resins, LR White and LR Gold, were tested for their suitability for post-embedding immunocytochemistry on human platelets. Both resins gave satisfying results with regard to ultrastructural preservation and retention of antigenicity for von Willebrand factor, fibrinogen, fibronectin and clathrin. Embedding in LR White and polymerization at 50 degrees C without accelerator proved to be the best method because it provided clear negative membrane contrast, good immunocytochemical localization of antigens and the entire procedure required a comparatively short time. Label for von Willebrand factor was found in typically eccentric location in alpha-granules, but also in the form of small caps directly outside the granule membranes, beneath the outer plasma membrane and in the cytoplasm. With the exception of the eccentric location within granules, labelling for fibrinogen and fibronectin revealed very similar results. Label for clathrin appeared in the form of small gold clusters attached to alpha-granules, in the cytoplasm and beneath membranes of the open canalicular system. Together with the appearance of coated vesicles attached to granules and to the membranes of the open canalicular system in araldite-embedded material, these results further strengthen the concept that adhesive proteins are taken up from the plasma and stored in granules. Moreover, the persistence of the clathrin coat on vesicles attached to storage granules may also indicate that receptors for adhesive proteins are recycled.
21 patients with severe active lupus nephritis (LN) were treated with intravenous cyclophosphamide monthly doses 0.75 g/m2. The effects were compared with the results obtained in 22 patients by the oral prednisone administration in the dose 1 mg/kg/day. Both groups were not significantly different as regards the initial intensity of the LN symptoms. Complete or partial remission occurred in 17 patients (80.9%) receiving intravenous cyclophosphamide. End-stage renal failure in 2 patients and moderate renal insufficiency in other 2 patients from that group. In the prednisone treated group complete or partial remission was observed in 13 subjects (59.1%). The significant deterioration of the renal function occurred in 9 patients, in 6 into the phase of a moderate renal insufficiency and in 3 patients to the end-stage kidney failure. Additionally, the significant increase of the total serum complement activity and of the plasma platelets count was observed in the cyclophosphamide group, whereas those indicators did not improve in the patients receiving prednisone. The of intravenous pulse cyclophosphamide exhibits an advantage in the treatment of severe proliferative LN.
In the present study influences leading to in vitro formation of dysmorphic erythrocytes were studied. Fresh and surface altered erythrocytes pretreated with several proteases were passed through different solutions representing distinct nephronís fluids. After passage of intact erythrocytes only 12-15% dysmorphic cells were observed. In case of protease treated cells the number of dysmorphic cells rose to 35-80%. The hemoglobin content was decreased. PAGE and electron microscopical findings demonstrated substantial losses of transmembrane and membrane skeleton proteins. It is assumed that surface protein degradation, loss of membrane skeleton proteins and hemolysis seem to be closely associated with dysmorphic malformation of urinary erythrocytes characteristical for glomerulonephritis.
We examined the effect of white cell contamination on thrombocytes prepared from pooled buffy coats over a storage period of 8 days. Using this novel technique, a leukocyte depletion filter can be easily integrated during PC preparation. In a paired study (n = 14) eight ABO-identical BC were pooled in a 2-liter PVC bag within 8 h after whole-blood donation, thoroughly mixed and divided into two identical fractions. After soft-spin centrifugation the platelet-rich plasma (PRP) was transferred either (fraction A) using a leukocyte filter (PL 50-HF, Pall) or (fraction B) directly into the storage bag (Pl-732, Baxter), and stored under routine conditions. On days 1, 3, 5, and 8, aliquots of PC were withdrawn for determination of cell count and different biochemical parameters and for morphometric analyses of platelet ultrastructure by electron microscopy. Results showed a lower thrombocyte yield and white cell count (p < 0.01) in fraction A (268 x 10(9) vs. 240 x 10(9); 51.1 x 10(6) vs. 0.04 x 10(6)), whereas no differences between the preparations could be detected by analysis of pH, pCO2, bicarbonate, and in LDH release over the storage period of 8 days. These results were supported in the study on the ultrastructural level where a good morphological integrity of the platelets was observed during the whole storage period in both fractions. In conclusion, storage lesions on platelets due to leukocyte effects are unlikely to occur in PC with white cell counts lower than 10(8)/l.
Both the investigation of the enteric nervous system and the diagnosis of its pathological changes require reliable staining methods. In order to assess the potential of protein gene product (PGP) 9.5 as a marker for the enteric nervous system, we examined its immunoreaction in whole-mount preparations of the guinea pig and porcine small intestine, using a rabbit polyclonal antiserum. The immunohistochemical technique reveals the fundamental architectural features of the ganglionic and aganglionic plexuses. Furthermore, it enables a reproducible and differentiated visualization of the enteric nerve cells to be made, so that the various nerve cell types can be morphologically identified.
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The use of implants to stabilize fractured diaphyseal bone, to handle difficult bone damage and to perform augmentation or replacement procedures in bone has become a common method in bone surgery. In most cases metal implants were used. Biodegradability of implant materials offers new perspectives. Restoration of the physiological status in the implant site becomes possible. Allergic reaction and second operations to remove the implants can be avoided and transitional aid in wound healing by the use of biomaterials can be achieved. An in-vitro system was established to investigate the interactions between osteoclasts and biomaterials, since it is the osteoclasts which are potentially able to resorb or degrade implants. The cell's resorption capabilities as well as its morphological behavior were documented. Two biodegradable and four nonbiodegradable materials were tested. The non-degradable materials provoked specific cell behaviour patterns but were not resorbed. Fibrin tissue adhesive sealant, however, displayed resorption lacunae mediated by osteoclasts, whereas polydioxanone (PDS) showed no resorption sites but normal cellular morphology when compared to the standard control (cells on hydrophilic coated teflon dishes). Both materials appeared to be well accepted by osteoclasts. This test system was established for the valuation of biodegradable implant materials and can be used to characterize new materials concerning their resorbability and biocompatibility without superposition by other cell systems.
Some modified cerium-based and Gomori-based cerium methods for the demonstration of phosphatase activity in cryostat sections were described. Dextrane as stabilizing agent was added to the incubation media for ATPase, 5'-Nase, and TPPase. The oxidation of the CeIII-phosphate primary reaction product in a separate step by H2O2 before the DAB incubation yielded an increase of the intensity of the DAB-based visualization reaction (Ce-H2O2-DAB-Ni two step method). The sensitivity of the histochemical enzyme reaction was remarkably increased if CeIII-ions were employed as amplifying agent (Ce/Ce-H2O2-DAB-Ni two-step method). A new suitable DAB medium consisting of 0.015% DAB, 2.0% Ni-sulphate, 15% methanol, and 0.005% H2O2 in 0.1 mol/l acetate buffer, pH = 5.2, was used. The disadvantage of diffuse background staining has been overcome by addition of 15% methanol to the DAB solution. Electrovalently bound CeIII (cerophilia) was removed by treatment of the incubated sections with CeIII-citrate (CeIII-complexation). In addition, a novel membrane floating incubation for sections is proposed. At present, the modified procedures are some of the most sensitive modes for the demonstration of phosphatases and improve the earlier described cerium-DAB one-step technique (Halbhuber et al. 1988b).
Two patients presented with multicystic lymphatic malformations of the penis. These were congenital and progressive, and both were successfully treated by excision after injection of methylene blue.
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