Wolf-Dieter Kuhlmann

MVZ für Laboratoriumsmedizin Koblenz-Mittelrhein, Koblenz, Germany · Immunology, Serology

Animal studies have shown stem-like properties of adult hepatocytes and stem-like properties of biliary epithelial cells with the latter originating from the canaliculi of HERING. In partial hepatectomy and after carbon tetrachloride injury, adult hepatocytes are the main source for liver restitution. In these cases, special stem cells are not requ...

Background: Serologic RhD typing may be unreliable for purposes in blood transfusion and pregnancy surveillance as monoclonal anti D reagents can react in full strength with weak and partial D types. We were close on false serologic RhD typing in things like that because agglutination tests reacted with high strength in different assays of a pregna...

Adult hepatocytes and liver-cell progenitors play a role in restoring liver tissue after injury. For the study of progenitor cells in liver repair, experimental models included (a) surgical removal of liver tissue by partial hepatectomy; (b) acute injury by carbontetrachloride; (c) acute injury by d-galactosamine (GalN) and N-nitrosomorpholine (NNM...

Cardiac operation produces substantial alterations within the immune system, which possibly predispose postoperative complications. However, the interplay between proinflammatory and antiinflammatory reactions and the cells involved in this process are not completely clear. Therefore, we investigated serum levels, as well as synthesis patterns, of...

Cardiac surgery with cardiopulmonary bypass (CPB) is known to induce an immune response whose nature has been increasingly elucidated during the recent decade. Clinically, patients usually show two to three of the four symptoms, which define the so-called systemic inflammatory response syndrome (SIRS). In addition, all parameters of the innate, non...

Polymorphonuclear leukocytes contain a large number of enzymes and bactericidal proteins stored in granules. Neutrophil activation induces degranulation and immediate release of these bioactive substances, including human neutrophil elastase (HNE) also known as elastase-2 (ELA2), which may contaminate whole blood units and blood components. The HNE...

Established parameters, e.g. C-reactive protein (CRP), do not differentiate specifically enough between patients developing an infection and those exhibiting an acute phase response following cardiac surgery. The objective of this prospective study was to investigate if procalcitonin (PCT) is more helpful than CRP. During a 1-year period, seven out...

In the years 1991-1993 the diphtheria antitoxin titres of the blood serum of 2404 persons living in germany was determined by means of the ELISA test. Its aim was to to get an idea of the population's protection given by vaccination against diphtheria. It became evident that the average protection given by vaccination against diphtheria was on the...

Diphtheria antitoxin antibodies were measured by solid-phase enzyme-linked immunosorbent assay (ELISA) on the basis of a calibrated antibody standard. Protective antibodies were expressed as International Units per ml (IU/ml). The method of quantitation employed is easily performed under routine conditions. Sensitivity proved to be 0.09 mIU/ml, and...

In tetanus immunization, the need for a booster vaccination can be easily determined by a serological analysis of tetanus antitoxin. Vaccination without knowledge of the immune status carries a high risk of postvaccinal complications. We have used the Pro. M.D. expert system shell to produce the knowledge base TETANUS, to improve the diagnostic int...

Prevalence of antibodies against tetanus toxoid (tetanus antitoxin titre) was measured in 5,235 males aged 17-60 years, 3,069 (59%) aged 20-22 years. Taking as criterion a threshold value of 0.1 IU/ml, 5,071 (97%) had adequate tetanus immunity. 1,301 of 4,355 aged 21-30 years (30%) actually had antibody concentrations above 6.3 IU/ml. Booster injec...

Vaccination without the knowledge of the immune status run the risk of complications to vaccination to a high degree. Therefore a quantitative serological detection of TETANUS-antitoxin is recommended before booster-vaccination. The diagnostic interpretation of results can be improved by including extensive knowledge using a neural-network-system i...

Um die Vorhersagbarkeit von Impfkomplikationen nach Tetanus-Auffrischimpfungen zu uberprufen, wurde die Antikorperpravalenz gegen Tetanus-Toxoid (Tetanus-Antitoxin-Titer) bei 5235 mannlichen Probanden im Alter von 17-60 Jahren bestimmt. Die meisten Manner (3069 [59 %]) waren zwischen 20 und 22 Jahre alt. Ausgehend von einer Schutzschwelle von 0,1 I...

Zusammenfassung Impfungen ohne Kenntnis des Impfstatus bergen in hohem Maße das Risiko von Impfkompli-kationen, weshalb sich vor einer Auffrischimpfung gegen Tetanus eine quantitative Messung von Tetanus-Antitoxin empfiehlt. Eine diagnostische Interpretation der Ergebnisse konnte durch den Einbezug umfangreichen Wissens für den einsendenden Arzt mi...

Vaccinations without knowledge of the immune status carry a high risk of postvaccinal complications. For this reason a quantitative serological determination of tetanus antitoxin is advisable before a booster vaccination is performed. The diagnostic interpretation of the laboratory results can be improved by the inclusion of extensive information g...

Determination of PMN-elastase is a laboratory parameter for determining the participation of neutrophilic granulocytes (polymorphonuclear leukocytes) in inflammatory reactions. In the study presented here we compared three immunoassay systems for the determination of PMN-elastase: two commercially available assay systems (the EIA-2h and IMAC assays...

Tetanus antitoxin in human sera was detected with solid-phase immunoassays in microtitration modules coated with tetanus toxoid by using Eu(3+)-labeled anti-human monoclonal antibodies on the basis of an exactly calibrated antibody standard. The use of a time-resolved fluorescence immunoassay (TR-FIA) significantly improved the quantitative detecti...

Tetanus antitoxin was measured by solid-phase enzyme-immunoassay in sera of 4187 men and 408 women in the age of 19 to 90 years. When 0.1 IE/ml antitoxin antibodies were supposed to be protective, then vaccination proved sufficient in 96% of men and 71% of women. Immunity to tetanus was at a higher level in the younger persons than in the older one...

The expression, inducibility, and regulation of four different cytochrome (cyt.) P-450 isoenzymes (PB1, PB2, MC1, and MC2) NADPH-cytochrome P-450 reductase, the glutathione transferases (GSTs) B and C and microsomal epoxide hydrolase (mEHb) have been studied during nitrosamine-induced hepatocarcinogenesis using immunohistochemical techniques. The i...

A sensitive staining procedure for glucose oxidase (GOD) as marker in immunohistology is described. The cytochemical procedure involves a two-step enzyme method in which GOD and horseradish peroxidase (HRP) are coimmobilized onto the same cellular sites by immunological bridging or by the principle of avidin-biotin interaction. In this coupled enzy...

Universal, polyclonal and monoclonal immunoperoxidase staining kits from BioGenex, Dako and Ortho were employed for the localization of antigens such as gastrin, prostate specific antigen, IgA, IgG, AFP and CEA in histological sections from formaldehyde fixed and paraffin embedded human specimens. The kit components were controlled by immunohistolo...

Nitrosamine-induced hepatocarcinogenesis has been used to investigate the regulation and expression of different drug-metabolizing enzymes in preneoplastic and neoplastic lesions in the female Wistar rat. The enzymes investigated were two phenobarbital-inducible cytochrome P-450 (cyt. P-450) isoenzymes (PB1 and PB2, mol. wt. 52 000 and 53 500, resp...

Universal, polyclonal and monoclonal immunoperoxidase staining kits from BioGenex, Dako and Ortho were employed for the localization of antigens such as gastrin, prostate specific antigen, IgA, IgG, AFP and CEA in histological sections from formaldehyde fixed and paraffin embedded human specimens. The kit components were controlled by immunohistolo...

Two cytochromes P450 (PB1 and PB2) have been isolated from the livers of rats treated with phenobarbital. PB2 (mol. wt. 53 500) is novel and is the first example of a phenobarbital-inducible enzyme with a Soret peak at 447 nm. Using an enzyme-linked immunosorbent assay, some immunochemical and structural similarities were observed between these cyt...

The histological localisation of alpha-D-galactopyranosyl residues in glycoconjugates of rat stomach and duodenal mucosae was studied by use of Griffonia simplicifolia agglutinin I, i.e. the isolectin mixture (A + B) and the isolectin B4 (B4). Cryostat sections which were either unfixed or acetone fixed and paraffin sections from both ethanol-aceti...

Peroxidase (HRP) conjugates ofDolichos biflorus agglutinin (DBA),Arachis hypogaea agglutinin (PNA) andUlex europaeus agglutinin I (UEA I) were used as specific molecular probes in order to describe histological patterns of carbohydrate moieties in glycoconjugates of gastric mucosa. Specimens of normal mucosa, metaplasias and carcinomas were fixed,...

Summary Four lectin-peroxidase conjugates (DBA, PNA, BSA I, UEA I) were used to stain intracellular carbohydrates in normal mucosa of human stomach and duodenum. Heterogenous patterns were observed with each of the lectins in a given cell population.

Peroxidase (HRP) conjugates of Arachis hypogaea agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Ulex europaeus agglutinin I (UEA I) and Bandeiraea simplicifolia agglutinin I (BSA I) were used to detect the expression of carbohydrate moieties in glyco-conjugates of normal human duodenal mucosa. Lectin histology was performed on sections from p...

Peroxidase (HRP) conjugates of Dolichos biflorus agglutinin (DBA), Arachis hypogaea agglutinin (PNA) and Ulex europaeus agglutinin I (UEA I) were used as specific molecular probes in order to describe histological patterns of carbohydrate moieties in glycoconjugates of gastric mucosa. Specimens of normal mucosa, metaplasias and carcinomas were fixe...

Procedures for ultrastructural postembedment immunolocalization of antigens were investigated by use of the indirect peroxidase labelled antibody method. Results were compared with those obtained with the ultrastructural preembedment technique. IgG globulins in IgG synthesizing cells of rat lymph nodes served as model. Formaldehyde fixation, ethano...

Cellular alpha 1-fetoprotein (AFP) is localized by light and electron microscopic immunoperoxidase methods. For light microscopic studies we employed ethanol-acetic acid fixed and paraffin embedded liver blocks. For the ultrastructural localization of AFP, formaldehyde-glutaraldehyde fixed tissues were used in preembedding phase. The occurrence of...

Various procedures for nonpolar and polar resin embedment were applied to mouse and rat livers for the study of postembedment immunolocalization of alpha 1-fetoprotein, albumin and the microsomal enzyme epoxide hydrolase. Fixations with formaldehyde and with formaldehyde-glutaraldehyde mixtures were used for tissue stabilization. Both fixation sche...

Alpha1-fetoprotein (AFP) was detected by serological, light and electron microscopic methods in various experimental models. These included (a) liver regeneration after partial hepatectomy or CCl4 intoxication (mouse and rat); (b) liver intoxication by high doses of N-nitrosomorpholine (NNM) and chemical induction of hepatomas (rat). AFP levels var...

Treatment of rats with N-nitrosomorpholine (NNM) for 7 weeks led to a focal increase in liver microsomal epoxide hydrolase (EH) as early as 2 weeks after withdrawal of the carcinogen. This treatment also leads to hyperplastic nodules and liver tumors, but much later. At the same early time point, ATPase activity was decreased in the same islands. M...

Intraperitoneal injections of galactosamine-HCl in rats were followed by transitory elevations of serum alpha-fetoprotein (AFP) concentrations. These were associated with regeneration of the damaged liver; a maximum of serum AFP was reached on day 4 (2.080.67 g/ml). In sera of untreated rats, serum AFP levels were g/ml and no cellular AFP was detec...

Intraperitoneal injections of galactosamine-HCl in rats were followed by transitory elevations of serum alpha-fetoprotein (AFP) concentrations. These were associated with regeneration of the damaged liver; a maximum of serum AFP was reached on day 4 (2.08 +/- 0.67 microgram/ml). In sera of untreated rats, serum AFP levels were less than 0.1 microgr...

The development of α1-fetoprotein (AFP)-producing cells was studied in rat liver by ultrastructural immunoperoxidase cytochemistry. AFP was detected in fetal and postnatal hepatocytes, including cell stages which ranged from the fetal to the adult cell-type. Irrespective of the stage of hepatocyte maturation, AFP was localized in the perinuclear sp...

Serum AFP concentrations in normal adult BALB/c/J and in normal adult C3H/He mice were in the order of 0.6 μg/ml and 0.1 μg/ml, respectively. In BALB mice, AFP was localized in the cytoplasm of differentiated mono- and binucleated hepatocytes in centrolobular and intermediate zones of normal adult liver. No cellular AFP could be detected in liver s...

Germ-free and conventional rats were fed cholesterol or normal pellets and part of both groups were immunized repeatedly with horseradish peroxidase (HRP). Four days after the last injections all the rats were killed, aortas removed and prepared for immunocytology and chemical analysis. Free peroxidase was not detected in the arterial wall and in a...

Hepatomas were induced in rats by feeding low doses (6 mg/kg) or high doses (20 mg/kg) of N-nitrosomorpholine (NNM) for 12 and 6 weeks respectively. Necroses of adult hepatocytes and proliferation of oval-shaped cells occurred only at high doses of NNM. In parallel, early reappearance of alpha1-fetoprotein (AFP) in sera was observed in rats which w...

Hepatomas were induced in rats by feeding N-nitrosomorpholine. Nodules from an alpha1-fetoprotein (AFP)-producing hepatoma were taken and subcutaneously injected into syngeneic hosts. From the first generation, the hepatoma producing most AFP was selected for further transfers. Four weeks after transplantation, the AFP concentration in the host rat...

Two different fixatives were applied to human gastric mucosa for the study of antigenic marker substances. The first consists of 96% ethanol and 1% acetic acid (EA method), the second of 4% formaldehyde, 0.5% picric acid and 0.25% glutaraldehyde (FPG method). Samples of resected gastric specimens were fixed, dehydrated and cleared in benzene and em...

Two different fixatives were applied to human gastric mucosa for the study of antigenic marker substances. The first consists of 96% ethanol and 1% acetic acid (EA method), the second of 4% formaldehyde, 0.5% picric acid and 0.25% glutaraldehyde (FPG method). Samples of resected gastric specimens were fixed, dehydrated and cleared in benzene and em...

Six-, 12- and 20-week-old rats were fed N-nitrosomorpholine (NNM) at low concentrations (6 mg/kg/day) or high concentrations (20 mg/kg/day) for 6 or 12 weeks. Irrespective of the age of the rats, both NNM schedules resulted in development of hepatomas and during the early stages of hepatoma induction, liver histotoxic patterns depended only on the...

Rat erythrocytes and lymphoid cells were fixed by glutaraldehyde and encapsulated in bovine serum albumin plus rabbit IgG globulins for cryo-ultramicrotomy. A technical procedure is described by which endogenous peroxidases of erythrocytes in ultrathin frozen sections were detected by hydrogen peroxide and diaminobenzidine as hydrogen donor. Modifi...

Mice were immunized with horseradish peroxidase and injected with [3H] thymidine. Popliteal lymph node cells were submitted to electron microscopic immunocytochemistry and high resolution autoradiography in order to correlate antibody production and ability to undergo cell division at various stages of lymphoid cell differentiation. Antibody synthe...

Mouse alpha1-fetoprotein (AFP) was isolated from amniotic fluid by immunoadsorbent columns and preparative electrophoresis. Specific antibodies were isolated from monospecific hyperimmunsera by use of immunoadsorbents, and subsequently coupled with horseradish peroxidase. At the light microscopical level, purified antibody-peroxidase conjugates wer...

Lymphocyte differentiation and specific antibody synthesis were studied in the course of the secondary immune response of horseradish-peroxidase immunized rat popliteal lymph nodes by electron microscopic immunocytochemistry. From primary immunization, mature plasma cells containing specific antibody remained as long as 10 months. Antihorseradish p...

Various experimental conditions were tried to stain intracellular IgG globulins within popliteal lymph node cells of rats and rabbits by the use of specific antibodies and their Fab fragments labelled with peroxidase following two different procedures.Fixation with 4% formaldehyde/24 hr, 1% formaldehyde and 1% glutaraldehyde/1–1.5 hr and 1–1.5% glu...

A method is described by which tissue blocks and cell cultures can be rapidly encapsulated for cryoultramicrotomy. Bovine serum albumin, insolubilized with glutaraldehyde, has been found to be a good embedding medium for biological purposes. Ultrathin frozen sections with large fields that are sufficiently uniform for low and high magnification mic...

The development of antihorseradish peroxidase-producing cells was studied by electron microscopic immunocytochemistry in rat popliteal lymph node after a single immunization of antigen with complete Freund's adjuvant. Specific antibody synthesis could be observed during the maturation stages of two morphologically distinct cell types from the blast...

A new staining procedure for glucose oxidase at both the light and the electron microscopic level has been developed. The procedure was equally effective for the detection of galactose oxidase. It appears that the method can be applied to all oxidoreductases which catalyze the production of hydrogen peroxide. Aspergillus niger glucose oxidase and D...

Different methods of fixation and tissue processing were employed to demonstrate intracellular antibody to horseradish peroxidase, Escherichia coli alkaline phosphatase and glucose oxidase in the popliteal lymph node of the rabbit, rat, guinea pig, and mouse. Fixation with 2.5% glutaraldehyde for 1.5 hours, 1.25% glutaraldehyde and 1% formaldehyde...

Mit monospezifischen Heteroimmunsera, die gegen die organspezifische antigene Esterase VI A gerichtet waren, wurden mit dem indirekten Nachweisverfahren nach Coons unter Verwendung FITC-chromierten Antiglobulinserums Gefrierschnitte der normalen Korpusschleimhaut untersucht. Aus dem Vergleich der histologischen und immunfluorescenzhistologischen Un...

1. Freie Radikale Freie Radikale sind sehr reaktive, instabile Atome oder Moleküle mit einem (oder mehreren) unge-paarten Elektronen (e −). Die Bezeichnung "Reaktive Sauerstoff-Spezies" (ROS) ist eine Sammel-bezeichnung und hierzu zählen z.B. • Freie Radikale: Hyperoxidanion, Hydroxylradikal, Singulett Sauerstoff, Stickoxid. • Nicht-Radikale O 2 De...

Gesundh.-Wes. 56, 667-671, 1994 Zusammenfassung In den Jahren 1991-1993 wurden aus den Blutseren von 2404 in Deutschland lebenden Personen die jeweiligen Diphtherie-Antitoxin-Titer im ELISA-Verfahren bestimmt, um sich ein Bild vom Impfschutz der Bevölkerung gegen Diphtherie machen zu können. Es zeigte sich, daß der durchschnittliche Impfschutz gege...

Ein personenbezogenes Analysenprogramm umfaßt die Messung von Spurenelementen, Antioxi-dantien und oxidativem Stress. Dabei lassen sich mit sog. Stress Checks oder mit gezielten Einzel-anforderungen antioxidative Versorgung, Entgiftungskapazität und oxidative Belastungsparameter beurteilen. Die Einzelanforderung ist vor allem für die Differentialdi...

Zusammenfassung Tetanus-Antitoxin wurde unter Verwendung eines Festphasen-Enzymimmunoassays im Serum von 4187 Männern und 408 Frauen im Alter zwischen 19 und 90 Jahren gemessen. Unter der Annahme einer protektiven Schutzschwelle von 0,1 IE/ml Antitoxin-Antikörpern ließ sich bei 96% aller Männer und bei 71% aller Frauen ein ausreichender Impfschutz...

Zusammenfassung Die Bestimmung der Elastase aus neutrophilen Granulozyten (PMN-Elastase) ist ein labora-toriumsdiagnostischer Parameter zur Erfassung der Abwehrreaktion neutrophiler Granulozy-ten. In der vorliegenden Arbeit wird die Leistungsfähigkeit dreier immunologischer Testver-fahren zur PMN-Elastasemessung verglichen. Es handelt sich dabei um...

Immunsystem Das Immunsystem dient der Aufrechterhaltung der körperlichen Integrität, die auf dem Prinzip der Selbst-Nicht-Selbst-Erkennung beruht. Wenn diese Unterscheidungsmöglichkeit aus dem Gleichgewicht gerät, entsteht Autoimmunität mit dem Ergebnis eines Zusammenbruchs von Toleranz gegenüber körpereigenen Strukturen. Unter physiologischen Bedi...