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Summary: Antigens of Candida albicans derived from both growth phases employing five different media and from five different strains using one medium were analysed by gel-electrophoresis and immunoblot-technique. No growth phase or growth-medium dependent differences were noted when commercially available rabbit anti-Candida or human antisera were employed. The SDS-PAGE patterns of the five strains analysed were quite similar. However, one strain isolated from a patient with proven systemic candidiasis showed an additional 60 kd membrane protein which was recognized only by human but not by rabbit antisera. The immunoblot-patterns of patient and blood donor sera were compared. A striking result was that patients sera specifically recognized a 43 kd soluble antigen, whereas blood donor sera did not. Interestingly, early sera of patients with systemic candidiasis showed an isolated IgM-response towards an 80 kd antigen of the soluble fraction. Our results suggest that tests based on these two antigens may facilitate the diagnosis of systemic candidiasis. Zusammenfassung: Wir verglichen durch Gel Elektrophorese und Immunoblot Antigen Prepa-rationen, die von funf verschiedenen Candida albicans Stammen sowie unter Verwendung von funf verschiedenen Anzuchtmedien gewonnen worden waren. Es konnten im Immunoblot keine von der Wachstumsphase des Organismus abhangige Unterschiede festgestellt werden. Die Protein-Bandenmuster der SDS-Gelelektrophorese waren ebenfalls vergleichbar. Bei der Analyse unterschiedlicher Stamme zeigte sich jedoch, das ein Isolat aus einer Blutkultur ein zusatzliches Antigen in der Membranfunktion aufwies, das nur von menschlichen, aber nicht von Kaninchen-Antiseren erkannt wurde. Weiterhin wurden die Immunoblot-Muster von Patienten und Blutspenderseren verglichen. Dabei zeigte sich, das Seren von infizierten Patienten mit einem 43kd Antigen reagierten, wahrend Blutspenderseren diese Reaktivitat nicht zeigten. Ein weiterer interessanter Befund war, das Patienten mit systemischer Candidainfektion in der Fruhphase IgM-Antikorperzeigten, die gegen ein 80 kd Antigen gerichtet waren. Unsere Ergebnisse lassen vermuten, das durch Verwendung dieser Antigene in einem ELISA-Test eine Verbesserung der Diagnostik systemischer Candidainfektionen erreicht werden kann.

Three days after the end of a bout of diarrhoea of 3 days' duration, a 19-year-old patient developed severe nocturnal thoracic pain unresponsive to isosorbitol dinitrate. There were no abnormal findings on physical examination, except a sweaty skin. SGOT (38 U/l), creatinine kinase (291 U/l, CK-MB 29 U/l) and lactate dehydrogenase (246 U/l) were all elevated. The ECG showed ST segment elevations in leads I, II, III, aVF and V1-V6 as well as negative terminal T waves in I, II, aVL, AVF and V3-V6, changes suggesting peri- and myocarditis. The Widal test gave a raised antibody titre (1:800) against Yersinia enterocolitica serotype O:3. Seven days later the immunoblot test demonstrated antibodies against the same organism, which was finally isolated from stool after 11 days. Treatment consisted of ciprofloxacin (500 mg twice daily for 14 days). All symptoms, as well as the biochemical and ECG abnormalities, quickly improved. The patient was discharged free of symptoms after 34 days.

Humoral antibodies can be induced in cotton topped (CT) marmosets with killed Herpesvirus saimiri (HVS) vaccines. The serum antibodies delayed, but did not prevent, malignant lymphoma in actively and passively immunized monkeys after inoculation with 10(3.8)TCID50 of cell-free HVS. Experiments are in progress to determine whether the vaccines are able to protect against smaller challenge dosages of HVS. The immunized monkeys were not resistant against transplantation of tumor cells. Infection with Epstein-Barr virus (EBV) and Marek's disease virus (MDV) did not protect against the tumors caused by superinfection with HVS. Attenuation of HVS was not achieved by passaging the HVS genome through different monkey species.

Wir untersuchten 165 Muttermilchproben auf das Hepatitis-C-Virus (HCV). Sie wurden von 143 chronisch HCV-infizierten Muttern gewonnen, die bei 152 Entbindungen 153 Kinder zur Welt brachten. Die Entnahme erfolgte zwischen dem 31. Tag vor und dem 384. Tag nach der Entbindung (Median: 4 Tage post partum). Keine der 165 Milchproben enthielt HCV-RNA. Wir zeigten, dass Bestandteile der Milch keine inhibitorische Wirkung auf den Nachweis des HCV mittels PCR haben. Dazu wurden 120 Proben mit jeweils einem Serum eines HCV-positiven und eines HCV-negativen Patienten versetzt. In den mit virushaltigem Serum versetzten Milchproben konnte in 79 von 99 Fallen ein positives PCR-Ergebnis gefunden werden. Von 134 Muttern stand ein innerhalb der Schwangerschaft oder bis zum 6. Monat danach (Median: 3 Tage post partum) gewonnenes Serum zur Bestimmung des Virustiters zur Verfugung. Neunundachtzig Frauen (66,4%) erwiesen sich als viramisch, die Viruskonzentrationen lagen zwischen 1×102 und 1×107 Kopien/ml (Median:6×103 Kopien/ ml). Von den 153 gestillten Kindern konnten wir 151 auf eine HCV-Infektion untersuchen. Dabei erwiesen sich 3 Kinder (2,0%) als infiziert. Diese Ubertragungsrate ist niedriger als die von 4,0% bei unserem bisher untersuchten Gesamtkollektiv von 624 HCVexponierten Kindern. Zwei der 3 HCV-positiven Kinder wurden bereits in utero infiziert, bei dem dritten Kind wurde das Virus spat in der Schwangerschaft oder unter der Geburt ubertragen. Diese Befunde zeigen, dass die chronische HCV-Infektion einer Frau keine Kontraindikation fur das Stillen darstellt.

Determination of hepatitis C virus (HCV) genotypes has become increasingly important during the last years for prediction of the clinical course and the outcome of antiviral therapy. Therefore, numerous different methods have been developed to enable HCV genotyping. However, many of them are very laborious and expensive, leading to limited usage in daily routine diagnostics. We have established a method which combines the speed of the new LightCycler technology with the use of amplification products generated for diagnostic quantitative HCV RNA determination. Differentiation of HCV genotypes is performed with these amplicons in a single step by using fluorophore-labeled hybridization probes. Although currently only two different acceptor fluorophores are available for the LightCycler, types 1, 2, 3, and 4, which are by far the prevailing HCV genotypes in Europe and the United States, can be distinguished. Genotypes of specimens from 190 chronically HCV-infected patients were determined by the LightCycler method and compared with the results of nucleotide sequencing. Concordant results were obtained for all samples. This new method offers a fast and convenient possibility to determine the quantitative HCV RNA load and the genotype in large-scale settings within about 4 h.

Ziel dieser Untersuchung: Darstellung der (1) Pravalenzen von Hepatitis B (HBV), Hepatitis C (HCV) und HIV Infektionen bei Drogentodesfallen in Hamburg von 1985 bis 1997, (2) Mehrfachinfektionen, (3) Leberhistolgie von 100 autopsierten Drogentodesfallen. Desweiteren werden epidemiologische und forensische Aspekte diskutiert. 1469 Drogentodesfalle wurden in diesem Zeitraum registriert: (1) Die Gesamtprevalenzen stellen sich wie folgt dar: HBV 35% (1997: 39%); HCV 51% (1997: 58%); HIV 7% (1997: 15%). (2) Mehrfachinfektionen sind haufig (28%): 19% aller Drogentodesfalle wiesen eine HBV + HCV Infektion auf. (3) Leberhistologie: eine Hepatitis wurde in 74% diagnostiziert und war haufig assoziiert mit einer HBV und/oder HCV Infektion (77% aller feingeweblichen Hepatitis-Diagnosen). 9% zeigten eine unauffallige Leberhistologie. Rechtsmedizinische Obduktionen und das Monitoring von HBV, HCV und HIV Infektionen bei Drogentodesfallen stellen ein wichtiges Instument dar, um Informationen uber den Einzelfall und die Situation bei i.v. Drogenabhangigen zu erhalten.

In einer prospektiven Studie wurde die vertikale Ubertragungsrate des Hepatitis-C-Virus (HCV) von Muttern, die wahrend Schwangerschaft und Entbindung HCV-infiziert waren, auf ihre Kinder untersucht. In das Studienkollektiv wurden 90 Kinder von 85 bekannten, chronisch HCV-infizierten Muttern aufgenommen. Von neun Muttern war eine gleichzeitige Infektion mit dem HIV-1 bekannt. Die Serumproben der 90 Kinder wurden innerhalb von acht Wochen nach der Entbindung erstmals untersucht; Nachuntersuchungen erfolgten uber mindestens drei Monate. Drei der 90 Kinder wurden HCV-infiziert (3,3%), nachgewiesen durch eine Antikorperpersistenz uber mindestens zwei Jahre sowie den Nachweis von HCV-RNA in der PCR. Eines der Kinder hatte leicht erhohte Transaminasen, war aber klinisch wie die beiden anderen Kinder auch unauffallig. 54 der Kinder wurden so lange nachuntersucht, bis die mutterlichen Antikorper nicht mehr nachweisbar waren; dies war im Schnitt nach einem Jahr der Fall.

Objective: Various laboratory methods for detection of cytomegalovirus (CMV) are available, but all suffer from limited sensitivity and specificity or are prone to sampling errors.

The risk of vertical transmission of the hepatitis C virus (HCV) from infected mothers to their children during pregnancy and delivery was determined in 120 children born to HCV-positive mothers. Methods included enzyme immunoassay and immunoblot for detection of HCV antibodies and reverse transcription polymerase chain reaction (RT-PCR) for detection of viral RNA. Six (5%) children were perinatally infected with HCV as shown by RT-PCR. None of the infected children had clinical signs of hepatitis. None of the pregnancies was complicated by abortion, stillbirth, premature birth, or malformation of the child. Special concern was given to the possibility of HCV transmission via breast milk. In no breast milk sample obtained from 34 HCV-infected mothers was HCV RNA detected. These observations indicate that HCV infection is not necessarily a contraindication for breast-feeding.

The reverse transcriptase V207I mutation within the hepatitis B virus (HBV) polymerase is associated with resistance to lamivudine in vitro. The prevalence of this mutation in treatment-naïve patients was 1% (1/96). A follow-up of the patient carrying this mutation prior to treatment revealed no loss of sensitivity of HBV to lamivudine in vivo.

Human cytomegalovirus (HCMV) is a major pathogen in newborn children and immunosuppressed individuals. There are various diagnostic methods for HCMV, but all test systems either suffer from limited sensitivity and specificity or are prone to sampling errors. Due to its unequaled sensitivity, strong hopes were pinned on the polymerase chain reaction (PCR) for the early diagnosis of active HCMV infections. However, when monitoring immunocompromised patients, unwanted positive results are often obtained without further evidence of active HCMV infection. To better discriminate between latent and active infections, different methods for the quantitation of viral genomes in clinical samples by PCR have been devised in the past years. Here we discuss the current standing of quantitative PCR in HCMV diagnostics.

This study demonstrates the dynamics in the epidemiology of hepatitis C virus subtypes. Subtypes 3a and 4a have become increasingly prevalent in patients where an infection within recent years can be assumed. Evidence is presented that the subtypes observed among younger patients can spread rapidly and lead to significant changes in the subtype distribution.

In this report we present some of the biochemical properties of the enzyme, here called pp28(PTK), isolated from particulate fraction of rat spleen (1). The kinase is very susceptible for polyions as regulators of the enzymatic activity. The polyanions like dextran sulfate or heparin inhibited, and polycations such as spermidin, protamin, poly-L-lysine and some random polypeptides containing tyrosine besides a basic amino acid, stimulated the enzyme markedly. The kinase showed high sensitivity towards class IA salts. In the casein phosphorylation reaction the apparent Km value for ATP was 4 microM. An unusual property is associated with autophosphorylation which leads to a reduced activity towards external substrates. Some kinase inhibitors described in the literature were tested for their potency.

To determine whether the persistent presence of antibodies to recombinant antigens of the hepatitis C virus (HCV) corresponds to the presence of hepatitis C virus RNA in the same serum, 85 anti-HCV positive patients were studied by the polymerase chain reaction (PCR). The focus of the research was on patients with chronic hepatitis. Eighty- three patients were found to be positive by PCR; only two were negative. In addition, liver biopsies taken from seven patients positive for anti-HCV were shown to contain HCV-specific RNA. Sera collected from three patients suspected to have NANB hepatitis on the basis of clinical symptoms were negative both for HCV antibodies and HCV RNA. The correlation between HCV antibody positivity and detection of HCV RNA was 97.6%.

Ein 14-jahriger Junge stellt sich wegen hochakut einsetzender allgemeiner Schwache, Kopfschmerzen, Ubelkeit und Brechreiz in der Notaufnahme einer Hamburger Klinik vor. Weiterhin klagt er uber Brustschmerzen beim Atmen und ein Gefuhl von Luftnot. Noch in der Notaufnahme kommt es zu einer schnellen Zunahme der Atemnot bis zur vollstandigen respiratorischen Insuffizienz. Zusatzlich fallen weite lichtstarre Pupillen auf. Der Patient wird wegen der fortgeschrittenen Atemlahmung sofort intubiert, maschinell beatmet und auf die Intensivstation einer Kinderklinik verlegt. Unter der Verdachtsdiagnose einer zerebralen Blutung wird ein kraniales CT veranlasst. Die erste Anamnese ist unauffallig, kein Trauma erinnerlich und keine Reiseanamnese.

Of the various forms of chronic viral hepatitis, in Germany 60-70% are caused by the hepatitis C virus (HCV). The virus arrives inconspicuously, i.e. an acute infection only leads to an increase in transaminases in 40% of cases and to an increase in bilirubin in only 20%. However, approximately 90% of infections take a chronic course and in 20% this leads to cirrhosis after only 20 years. The infection rate of medical personnel is not significantly higher than in the general population. The transmission of HCV from patients to medical personnel, e.g. by needle stick injuries, is very rare and the risk of infection is less than 1%. Even less frequently transmission of HCV in the reverse direction from medical personnel to patients occurs. An active or passive prophylactic immunization is not possible and protective immunization is not yet foreseeable. Recently, progress has been made with chemotherapeutical treatment of HCV. The present state-of-the-art is pegylated interferon-a in combination with ribavirin. The success rate in HCV genotypes 2 and 3 is clearly higher with 70-80% than in genotypes 1 and 4 with approximately 40%. Both drugs have significant side-effects but better forms of medication are not yet available.

summary. The prevalence of a newly described DNA virus (SENV-H) was examined in a population of 599 individuals by polymerase chain reaction (PCR). All individuals were assigned to a nonrisk or a risk group depending on the presence of historical or serological factors indicating an increased risk for parenterally transmitted diseases. In a group of 226 healthy blood donors, 38 (16.8%) were found to be SENV-H viraemic. The highest prevalence of SENV-H viraemia was observed among patients infected by HIV (28 of 63; 44.4%). Contrarily, of 78 individuals on maintenance haemodialysis, only 10 (12.8%) were found positive in the SENV-H PCR. Our results demonstrate that SENV-H viraemia is widespread in the general population. Therefore, it seems to be questionable if parenteral transmission is the main route for spreading SENV-H. The hepatitis-inducing capacity of SENV-H is unclear. However, taking our clinical and epidemiological data into account it seems unlikely that this virus is responsible for hepatitis.

The HIV-status of 3999 fatalities (aged 1-60 years) examined at the Institute of Forensic Medicine in Hamburg from 1989-1992 was tested. Former predictions of an enormous increase of HIV-infections, especially in the risk group of IVDA (intravenous drug addicts), did not come true. HIV-screening of fatalities is an instructive additional method of gaining information about the epidemiological development of HIV-infections.

It remains unclear whether sequential assessment of hepatitis B virus (HBV) load during lamivudine therapy can predict the loss of hepatitis B e antigen or emergence of drug-resistant variants. Therefore, a longitudinal study was carried out in 28 consecutive patients with chronic hepatitis B who started lamivudine therapy for a median of 12 months (range, 6-31). HBV DNA copy numbers were determined at 3-month intervals. From month 6 onward, HBV viral load below the detection limit of the PCR was predictive of the loss of envelope antigen (P = 0.043). Continuously detectable HBV DNA during the first 12 months of treatment indicated emergence of drug-resistant variants (P = 0.034). These data suggest that the goal of lamivudine therapy should be complete suppression of serum HBV DNA.

A novel approach to predicting symptomatic cytomegalovirus (CMV) infections combines the level and the duration of viraemia in a single parameter. Sixty-four kidney transplant recipients were monitored by quantitative shell vial culture, pp65 antigenaemia, and polymerase chain reaction (PCR) of leucocytes. The area under the curve (AUC) of each parameter was determined from the onset of viraemia to the beginning of antiviral treatment. The AUC values were significantly higher in symptomatic than in asymptomatic patients. For antigenaemia and PCR, optimal AUC thresholds for predicting symptomatic CMV infections were determined. They were superior to standard cutoff levels of absolute viral load in sensitivity, specificity, and positive and negative predictive value. In 8 of the 23 patients who became symptomatic, impending clinical features were indicated earlier by the AUC thresholds than by standard viral load. In conclusion, the concept of the AUC should facilitate identification of patients at risk of symptomatic CMV infection.

To evaluate a new fourth generation assay for simultaneous detection of antibodies to the human immunodeficiency virus (HIV) 1 and 2 and HIV p24 antigen in daily routine we tested 675 sera obtained from 673 patients and compared the results to conventional antibody tests. In 546 uninfected patients the rate of unspecific reactivities was slightly higher in the new screening assay as compared to conventional antibody assays (1.1% vs. 0.4%). All 121 sera derived from patients with known HIV infection were detected correctly. In six patients from whom sera were obtained during early seroconversion the fourth generation ELISA was positive in three cases, while conventional third generation tests still were negative. In patients negative for HIV antibodies and low amounts of p24 antigen less than 100 pg/ml also the fourth generation ELISA remained negative. Thus, this new assay permits earlier detection of HIV infection and reduces the diagnostic window. It is a reliable tool for routine diagnosis of HIV, especially in blood donors and patients with high risk behavior.

The aim of the study was to examine whether the diagnosis of Hepatitis C (HCV) infection can be obtained reliably without using an immunoblot-based confirmation assay. 1,708 EIA-reactive serum samples were examined retrospectively for (i) optical density value in the screening assay, (ii) reactivity in an immunoblot assay, and (iii) result by RT PCR. In 1,394 (81.0%) samples positive results were obtained by both the HCV EIA and the confirmation assay. OD-valuesor = 2.2 were observed in 1026 of these samples, but covered the range from 0.4 to 2.1 in the other 368 samples. The combination of HCV EIA reactivity and indeterminate immunoblot assay was observed in 134 (7.8%) serum samples. HCV RNA was detected in 58 cases by PCR. The OD-values of these 58 samples ranged from 0.4 to2.2. Especially reactivity against the core recombinant protein was indicative of PCR positivity. The reactivity by the HCV EIA could not be confirmed by immunoblot assay or PCR in 180 (10.5%) sera. These false reactive sera showed OD values by EIA from 0.3 to 2.1. It is concluded that no threshold values can be defined which would allow differentiation between positive, indeterminate, and false reactive result by HCV EIA without producing an unacceptably high number of false negative diagnoses. Not using immunoblot-based confirmation would result in many additional PCR examinations. Therefore, confirmation of reactive HCV EIA results by a serological confirmatory assay must remain an essential part of the diagnostic procedure.

Known and potential virulence factors of enterococcal blood culture isolates were studied using 89 Enterococcus faecalis and 24 Enterococcus faecium isolates. The prevalence of the respective factors was (Enterococcus faecalis vs. Enterococcus faecium): hemolysin 16% vs. 0%, gelatinase 55% vs. 0%, aggregation substance 63% vs. 13%, lipase 35% vs. 4%, hemagglutinin 97% vs. 0%. Deoxyribonuclease was not detected in any isolate. The study showed that hemagglutinin and lipase may represent additional virulence factors of Enterococcus faecalis but not Enterococcus faecium. The significance of these factors in the pathogenesis of enterococcal infection needs to be elucidated in further studies.

In einer prospektiven Studie wurde die vertikale Ubertragungsrate des Hepatitis-C-Virus (HCV) von Muttern, die wahrend Schwangerschaft und Entbindung HCV-infiziert waren, auf ihre Kinder untersucht. In das Studienkollektiv wurden 90 Kinder von 85 bekannten, chronisch HCV-infizierten Muttern aufgenommen. Von neun Muttern war eine gleichzeitige Infektion mit dem HIV-1 bekannt. Die Serumproben der 90 Kinder wurden innerhalb von acht Wochen nach der Entbindung erstmals untersucht; Nachuntersuchungen erfolgten uber mindestens drei Monate. Drei der 90 Kinder wurden HCV-infiziert (3,3%), nachgewiesen durch eine Anti-korperpersistenz uber mindestens zwei Jahre sowie den Nachweis von HCV-RNA in der PCR. Eines der Kinder hatte leicht erhohte Transaminasen, war aber klinisch wie die beiden anderen Kinder auch unauffallig. 54 der Kinder wurden so lange nachuntersucht, bis die mutterlichen Antikorper nicht mehr nachweisbar waren; dies war im Schnitt nach einem Jahr der Fall.

A detergent extract isolated from the enriched fraction of integral membrane proteins of Jurkat cells showed an enhanced tyrosine phosphate level when phosphorylated in the presence of phorbol 12-myristate 13-acetate (TPA) and phorbol 12,13-dibutyrate (PDBu). The enhanced tyrosine phosphorylation was observed when the reaction time exceeded 6 min; at shorter incubation times, however, TPA inhibited tyrosine phosphorylation. When the reaction proceeded for a constant time period longer than 6 min and phorbol esters were added at different times after the start of the reaction, two phases of an enhanced tyrosine phosphorylation of a 50 kDa protein were observed. An increased phosphorylation of the 50 kDa protein was correlated with an enhanced phosphorylation of poly(Glu4,Tyr1). The two phases of enhanced phosphorylation differed in their TPA and PDBu requirement and in the proteins that were tyrosine phosphorylated. Studies with protein kinase C (PKC) inhibitors showed a negatively correlated effect on the enhanced tyrosine phosphorylation in phase I; tyrosine phosphorylation was further augmented. In phase II the regulation of tyrosine phosphorylation correlated with the efficiency of the PKC inhibitors on the alpha-isoform of PKC which was found in the cell extract. Separation of the proteins present in the investigated cell extract by gel filtration revealed a co-migration of the alpha-PKC and the 50 kDa protein. The metabolic labeling of intact Jurkat cells with 32Pi indicated that phorbol esters are also able to induce tyrosine phosphorylation of the 50 kDa protein underin vivo conditions. These data suggest an activation of two different tyrosine phosphorylation pathways by phorbol esters involving tyrosine phosphorylation/autophosphorylation of a 50 kDa kinase, as confirmed by 5'-p-fluorosulfonylbenzoyladenosine (FSBA) labeling, that are accurately regulated by alpha-PKC.

Staphylococcus caprae is a catalase-positive, coagulase-negative coccus that has been originally isolated from goat milk. We describe the first case of an intra-articular empyema caused by S. caprae in an immunocompetent patient following arthroscopic cruciate ligament repair. The patient recovered completely after debridement and antibiotic therapy with cefazolin and amoxicillin, respectively. This case demonstrates that this organism may rarely cause serious nosocomial infections in immunocompetent adults.

Das Hepatitis C Virus (HCV) ist ein etwa 9300 Basen langes plus-Strang RNA Virus, das mit den Flavi- und Pestiviren zur Familie der Flaviviridae gehort. Eine Infektion erfolgt in erster Linie uber eine parenterale Ubertragung. Nachdem spatestens seit dem 01.11.92 nach BGA Vorschrift Blut und Blutprodukte auf das Vorhandensein von Antikorpern gegen das Hepatitis C Virus hin untersucht werden mussen [1], liegt das Risiko eine HCV Infektion mittels Bluttransfusion zu erwerben bei 1:10000 bis 1:100000. Bei unserer Untersuchung der Infektionsrisiken bei 4659 HCV-Infizierten im Raum Hamburg stellten die intravenos Drogenabhangigen mit 23,4% (1090/4659) die groste bekannte Risikogruppe dar [2].

The detection of HCV-PCR amplification products by DNA enzyme immunoassay (DEIA) was compared with conventional hybridization carried out with a 32P-labelled oligonucleotide probe. The detection limit of both methods was shown to be between 100 pg and 1 ng of amplicon. All serum samples of 40 HCV-seropositive patients were positive after PCR in autoradiography, but only 38 with the DEIA technique (sensitivity 95%). There were no false-positive reactions by either method. The advantage of the DEIA method was the fast and non-radioactive detection of HCV amplicons. DEIA combines the specificity of the hybridization event with the speed of an ELISA procedure and is suitable for HCV-PCR.

The serodiagnosis of hepatitis C virus (HCV) infection was analyzed by a recombinant immunoblot assay (RIBA) with recombinant proteins encoded by the viral RNA isolated from our patients in Hamburg, Germany. The HCV RNA was amplified by PCR, and proteins encoded by the viral core and the NS3, NS4, and NS5 regions were expressed subsequently in Escherichia coli. The results obtained with our UKE RIBA were compared with the results of the Abbott HCV second-generation enzyme immunoassay (EIA). Serum samples from 270 patients, which were sent to us on the suspicion of HCV hepatitis and which were negative for hepatitis A virus and hepatitis B virus antibodies, were examined. In 227 cases (84.1%), there were identical positive (204 cases, 75.6%) or negative (23 cases, 8.5%) results in both tests. In 32 cases (11.9%), the reactive Abbott second-generation HCV EIA results could not be confirmed by the UKE RIBA and the HCV PCR. In follow-up studies conducted over 1 year, these results did not change. In three cases (1.1%), the UKE RIBA presented a positive result while the Abbott second-generation HCV EIA was negative. All three cases were positive in the HCV PCR and showed seroconversion in an HCV EIA 4 to 6 weeks later. In addition, 33 patient serum samples were examined by UKE RIBA in parallel with the Ortho RIBA 2.0. In three cases (9.1%), a positive Ortho RIBA 2.0 result could not be confirmed by the UKE RIBA and the HCV PCR. All three patients were free of complaints. The UKE RIBA showed also a smaller number of indeterminate results (3.0%) than the Ortho RIBA 2.0 (24.2%). This comparison study demonstrates that the commercially available HCV antibody tests should be further improved.

Virus infections are common among drug abusers and the medical complications are severe [1, 2, 3, 4]. Many infections are due to an insterile technique of injection (needle sharing); disease transmission via blood or body fluids may also be associated with sexual intercourse.

Compared with other nosocomial infections, transfusion-associated infectious risks are of minor relevance in the Federal Republic of Germany today. The actual spectrum of measures for an optimal degree of safety of blood components comprises a careful donor selection, highly sensitive and specific screening assays for viral parameters, inactivation and quarantine procedures for plasmatic preparations. Further possible improvements, both in the blood bank and clinical setting are discussed with regard to the inevitable residual risks of hemotherapy.

Hepatitis B virus-DNA was detected by polymerase chain reaction in 9 out of 10 patients after orthotopic liver transplantation. Three of these patients were at the same time positive for hepatitis B virus-DNA by dot-blot hybridization (hepatitis B virus-DNA1.5 pg/ml). In these three patients HBs-antigen (HBsAg) reappeared within a mean time of 12 weeks after orthotopic liver transplantation (range 7-18 weeks). Only two of the six polymerase chain reaction-positive and dot-blot-negative patients (hepatitis B virus-DNA between 0.4 fg/ml and 1.5 pg/ml) had recurrence of HBsAg within a mean time of 54 weeks (range 52-56 weeks). Passive immunoprophylaxis with anti-HBs antibodies (serum titers100 IU/l) did not prevent infection of the graft in the five reinfected patients. We conclude that a low concentration of serum hepatitis B virus-DNA after orthotopic liver transplantation, which is detectable only by polymerase chain reaction, indicates a delayed infection of the graft.

The in vitro activity of fleroxacin was compared to that of ciprofloxacin, ofloxacin and three broad-spectrum beta-lactams against 159 clinical isolates using the E-Test. Fleroxacin, ciprofloxacin, and ofloxacin were all active against Enterobacteriaceae with 93, 98 and 94% susceptibility, respectively. For other gram-negative strains, 81% were susceptible to fleroxacin and ofloxacin and 98% to ciprofloxacin. With regard to gram-positive strains, 72% were susceptible to fleroxacin, 83% to ciprofloxacin and 78% to ofloxacin. The broad antibacterial activity suggests that fleroxacin may be an alternative in the treatment of various infections, particularly in light of its favorable pharmacokinetics.

Der Nachweis von Antikorpern gegen das Hepatitis-C-Virus (HCV) [3] bedeutet, das sich mit uber 94%iger Wahrscheinlichkeit auch das HCV im Blut befindet, und das dieses bei parenterealer Ubertragung als infektios zu betrachten ist. Wir haben die HCV-Antikorper-positiven Serumproben von 652 Patienten in der PCR untersucht [4] und bei 617 (84%) das HCV nachgewiesen. Wenn das Serum nach der Blutentnahme rasch von den Blutzellen getrennt und in einer speziellen Transportlosung eingesendet wird, die den Abbau der sehr empfindlichen HCV-RNS durch Ribonukleasen im Serum erschwert, liegt die Nachweisrate sogar uber 94%. Die Konzentration der HCV-RNS im Blut kann im Verlauf der Erkrankung und von Patient zu Patient betrachtlich schwanken.

Chlamydia trachomatis was isolated from ten of 188 biopsies (5.3%) obtained from different parts of the lower digestive tract. Patients (mean age 37.0 years) presented with ulcerative proctitis, Crohn's disease, mild colitis or ulcerative colitis. Seven rectal biopsies, two biopsies from the sigmoid flexure and one caecal biopsy were positive for chlamydial isolation whereas all biopsies taken from the colon ascendens, transversum or descendens and from the terminal ileum were negative. We conclude that isolation of C. trachomatis is most effective from rectal and sigmoidal biopsies and is a rare event from other sites of the lower digestive tract.

Twenty-eight patients with different stages of HIV infection who had not undergone antiretroviral pretreatment were treated with an escalating dose regimen of zidovudine (4 weeks 2 x 50 mg, 4 weeks 2 x 100 mg, 4 weeks 2 x 250 mg). CD4+ cell counts and p24 antigen levels were monitored every four weeks. Twenty-one patients were evaluable. p24 antigen levels showed a significant decrease after four weeks (p0.01, Sign test, Wilcoxon matched pairs test) that was sustained until week 12 without a further significant decrease. CD4+ cell counts increased significantly within the first four weeks (p0.01, Sign test, Wilcoxon matched pairs test). This increase was sustained until week 12 but no further significant increase was noted. Mean corpuscular erythrocyte volume values increased significantly after week 4 and continued to rise until week 12. These results demonstrate antiretroviral activity of a very low zidovudine dose, however low doses should not be used for treatment unless the clinical efficacy is shown to be equivalent to that of standard doses.

A phosphoprotein with associated tyrosine kinase activity has been purified from a 26,000 x g particulate fraction of rat spleen. The molecular weight of the native kinase is 85-90 kDa as estimated by gel permeation chromatography on Superdex 200 while SDS/PAGE of an autophosphorylated sample indicated a single 28-kDa band in which the radiolabel was alkali-stable. The SDS/PAGE following the incubation with 5'-p-fluorosulfonylbenzoyl[8-14C]adenosine reveals a single labeled band migrating at a molecular weight of approximately 28 kDa. The enzyme shows very restricted substrate specificity. Casein and, to a lesser degree, the random polypeptide poly(Tyr1,Glu4) were phosphorylated. The enzyme exhibits a preference for Mn2+ over Mg2+ as an activator.

There is a distinct spectrum of infectious diseases in prisons, known as desmoteric infections. We investigated the incidence of HIV, hepatitis A, B, and C in a group of 539 prisoners and compared them with the incidences of these infections in the normal population. The incidence of HIV was 928 out of 100,000. This figure is similar to that of the normal population. Hence, HIV infection does not seem to be a desmoteric disease. In contrast to AIDS the incidences of hepatitis A, B, and C among prisoners were 2968, 1670, and 20,000, respectively. Of course, hepatitis A, B, and C occur 100-200 times more often among prisoners and thus seem to be desmoteric infections. Moreover, especially hepatitis C is correlated strongly with intravenous drug abuse. About 80% of drug abusers were HCV positive and 50% of the total of HC infections are due to drug abusers.

Three days after the end of a bout of diarrhoea of 3 days' duration, a 19-year-old patient developed severe nocturnal thoracic pain unresponsive to isosorbitol dinitrate. There were no abnormal findings on physical examination, except a sweaty skin. SGOT (38 U/l), creatinine kinase (291 U/l, CK-MB 29 U/l) and lactate dehydrogenase (246 U/l) were all elevated. The ECG showed ST segment elevations in leads I, II, III, aVF and V1-V6 as well as negative terminal T waves in I, II, aVL, AVF and V3-V6, changes suggesting peri- and myocarditis. The Widal test gave a raised antibody titre (1:800) against Yersinia enterocolitica serotype O:3. Seven days later the immunoblot test demonstrated antibodies against the same organism, which was finally isolated from stool after 11 days. Treatment consisted of ciprofloxacin (500 mg twice daily for 14 days). All symptoms, as well as the biochemical and ECG abnormalities, quickly improved. The patient was discharged free of symptoms after 34 days.