Biomedical subjects
Publications and source records attributed to N Müller.
With a new method which is easy to handle, leukocyte depleted neocyte enrichment can be obtained from ordinary whole blood units using centrifugation. In comparison to alternative methods less time for preparation will be needed without significant changes in enzyme activities during storage indicating red cell lesion.
The article elucidates the new laboratory tests for chronic joint diseases, with special attention to quantitative and qualitative serum markers for metabolic and immunologic processes. The assays include the quantification of keratan sulphate, hyaluronan and beta-D-xylosyltransferase in serum and pyridinolin in serum and urine as markers for cartilage catabolism, as well as the detection of autoantibodies to cartilage matrix and cell components and cartilage-directed T-cell reactions in the patients. Taken together, these assays represent models for future laboratory diagnostic tools for the characterization of arthritic diseases. So far, however, it remains unclear whether they will prove to be adequate assays for the practicing physician.
247 human cadaver jaws (obtained from 115 complete denture-wearers, 98 partial denture-wearers, 34 control persons) were examined histopathologically to determine if there is a correlation between tissue response and type of prosthetic management. The pathohistological changes of tissue and bone were evaluated statistically according to sex, age and topography of maxilla and mandible. The highest number of pathological findings was associated with those complete dentures and partial dentures where the biomechanical principles of construction had not been observed. Age, sex and topography, however, do not reveal any significant influence on tissue and bone.
We describe the use of the Escherichia coli plasmid vector, pVB2, for high-level expression and export of recombinant protein. The pBR322 derivative pVB2 harbors the mglB gene, which codes for the galactose-binding protein (GBP) of E. coli. GBP is exported into the periplasmic space of the bacterial cell. Gene mglB contains an EcoRI restriction site close to its 3' end which allows simple in-frame insertion of EcoRI fragments obtained from recombinant lambda gt11 phages. The pVB2 vector was used to express an antigen from Echinococcus multilocularis. The recombinant protein amounted to over 50% of total cellular protein and could be efficiently isolated from the periplasm by osmotic shock. The application of the purified antigen in an ELISA enabled a clear and specific detection of anti-Ec. multilocularis antibodies in human patients' sera, which had been immunosorbed with a periplasmic extract (containing wt GBP) before investigation. These data show the general usefulness of pVB2 as an expression vector for producing in E. coli diagnostically relevant antigens from any infective organism.
A highly antigenic polypeptide fragment of the recombinant Echinococcus multilocularis antigen II/3 was produced in Escherichia coli and purified for application in enzyme-linked immunosorbent assay (ELISA). The antigen II/3-encoding 1.0 kb DNA sequence was reduced by sonication into smaller DNA fragments which were subsequently cloned into lambda gt11. Three clones could be isolated from the sublibrary, all synthesizing a recombinant antigen as a stable beta-galactosidase fusion protein. In a further step, the 0.6-kb insert from one positive clone was subcloned into the plasmid pAR 3038, which directed efficient synthesis of the antigen fused to only 11 amino acids from the N-terminus of the phage T7 major capsid protein. The plasmid-encoded antigen (antigen II/3-10) was purified from a bacterial cell extract and then tested in an ELISA. Using sera from 88 patients with an E. multilocularis-infection, a high diagnostic sensitivity of 90% was demonstrated. Investigation of sera from 220 patients with various helminthic infections showed a specificity of 99%, suggesting the suitability of the antigen II/3-10 as an immunodiagnostic tool.
Several endocrine parameters were assessed in 35 alcoholic inpatients after admission to hospital, and 17 of the 35 were retested after several weeks of sobriety. No difference was found in clonidine-stimulated growth hormone (GH) secretion between male alcoholics and male healthy controls, but significant positive correlations of GH secretion and alcohol content in expired breath on admission and gamma-GT values after abstinence were observed. Nonsuppression in the dexamethasone suppression test was found in 17% of the patients on admission, which seemed to be due to alcohol withdrawal. Postdexamethasone cortisol levels were significantly positively correlated with the "apathic syndrome" (r = 0.40; p less than or equal to 0.05). About one-third of the patients showed a blunted response in the TRH-test both on admission and after abstinence. No significant influence of alcohol intake, withdrawal or familial disposition on prolactin values could be detected. The results of the TRH test and the DST point to similar endocrinological patterns in alcoholics as in depressive patients and thus support the hypothesis of a link between alcoholism and depression.
Uptake of bumetanide into rat liver cells was investigated using isolated hepatocytes and primary cell cultures. The kinetics of [3H]-bumetanide uptake revealed two saturable components in addition to an unsaturable component. Saturable bumetanide uptake consists of a high-affinity, sodium-dependent uptake and a low-affinity transport system. Bumetanide uptake into isolated rat hepatocytes is energy dependent and temperature sensitive. At low temperatures, bumetanide uptake is due to diffusion with a permeability coefficient of 1.16 x 10(-6) cm/s. In primary liver cell cultures, uptake of bumetanide decreases rapidly over 3 days. AS-30D ascites hepatoma cells do not take up bumetanide but bind small amounts of the loop diuretic. Hepatocytes metabolized bumetanide extensively. The metabolites were secreted into the surrounding incubation buffer. Two hydroxylated and at least one conjugated biotransformation product could be separated by thin-layer chromatography. Isolated rat hepatocytes possess carrier proteins for uptake of bumetanide and very likely also for uptake of other loop diuretics like furosemide, piretanide, and torasemide. Several inhibitors of multispecific transport systems in the kidney and liver were tested as potential inhibitors of hepatocellular bumetanide or furosemide uptake. Probenecid, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, iodipamide, digitoxin, bile acids, and bromosulfophthalein inhibited uptake of loop diuretics. Inhibition by taurocholic acid was competitive with a Ki of 24 microM. Taurocholic acid inhibited [3H]bumetanide uptake in the presence but not in the absence of Na+. Deoxycholic acid and bromosulfophthalein were noncompetitive inhibitors of hepatocellular bumetanide uptake.(ABSTRACT TRUNCATED AT 250 WORDS)
The adverse clinical potential of leucozytes in red cell and platelet transfusion became more widely recognized. Therefore, it is preferable to remove as many white cells as possible to avoid all these risks. The new filtration methods offer improved removal efficiency and leave a quantity of leucocytes that is below the threshold for induction of a febrile reaction in most sensitized patients and close to the threshold to avoidance of alloimmunization. A selective white cell removal from platelet concentrates using new filters, in which fibers were coated with a particular polymer, increases the efficiency of trapping white cells and also prevents platelet adhesion. These filters are capable of remove more than 95% of the leucocytes with minimal platelet loss, normal morphology and in vitro function. These filters can be used in bag-to-bag as well as in bedside filtration and greatly reduce the side effects caused by white cells in blood products.
23 Patients were followed up clinically and by X-ray. In this cohort eight maxillary and 19 mandibular alveolar bone augmentations with hydroxylapatite granules mostly in a subperiosteal tunnelling procedure had been done. In most cases prothetic treatment was administered in our hospital. The dentures had been in place for a little over three years. Over this period of time there was hardly any change in the shape of the augmented ridges, denture retention, and vertical dimension. Drawbacks of the procedure became evident as reduced load resistance of oral mucosa during mastication and recurring impingement spots. In some cases, particularly when there was little vertical bone height, pain occurred in the area of the mental nerve.
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Aflatoxin B1 permeates isolated rat hepatocytes by non-ionic diffusion. Its uptake is neither saturable nor influenced by metabolic energy and not inhibited by treatment of cells with proteases. The initial rate of aflatoxin B1 uptake measured at 7 degrees C is between 40 and 50% compared to that at 37 degrees C. However, after an incubation period of 7 minutes identical equilibrium uptake is reached at both temperatures. The apparent activation energies, calculated for aflatoxin B1 uptake by Arrhenius diagrams ranged between 1.69 and 4.5 kcal/mol. A Q10 value of 1.34 was calculated for a temperature interval of 7-17 degrees C but decreased to 1.05 for the interval of 27-37 degrees C. Liposomes or lipoproteins added to the cell suspension inhibited the aflatoxin B1 uptake into hepatocytes. Liposomes mainly composed of unsaturated fatty acids bind twice as much aflatoxin B1 as those composed of saturated ones, indicating that the lipophilicity of the mycotoxin is crucial in the determination of its uptake into liver cells. At concentrations above 5 micrograms/ml, aflatoxin B1 inhibited the carrier-mediated uptake of cholic acid and of phalloidin into hepatocytes. This effect was reversible and abolished by washing the cells after preincubation with aflatoxin. In concentrations below 5 micrograms/ml the uptake of phallotoxin and cholic acid was however stimulated by 15-25%. These results indicate, that a carrier-mediated uptake into hepatocytes via the multispecific bile salt transporter is not responsible for the organoselective clearance of aflatoxins by the liver. On the other hand, the cholestatic effect of aflatoxin B1 results at least partially from the inhibition of the multispecific bile acid transport system. This inhibition may arise from affinity of aflatoxins to lipid domains of the cell membrane.
A cDNA library derived from mRNA of metacestodes from Echinococcus multilocularis was constructed in the Escherichia coli expression vector lambda gt11 and screened with human patients' sera. The recombinant proteins of 11 positive phages were further evaluated for their potential as immunodiagnostic reagent. One isolated clone (from lysogen II/3) synthesized a fusion protein which was rapidly degraded. This intracellular degradation process provided two distinct polypeptides with Mr of about 31,000 and 33,000, both showing strong binding activity with specific patients' antibodies. The diagnostic value of the II/3-antigen was evaluated by mini-Western-blot with sera from 41 patients infected with E. multilocularis and sera from 77 patients with other helminthic infections, resulting in a diagnostic sensitivity of 98% and an over-all specificity of 96%. These results suggest the usefulness of the recombinant II/3-antigen for immunodiagnosis of human alveolar echinococcosis.
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