Shiga toxin-producing Escherichia coli (STEC) shedding in a wild roe deer population.

Worldwide infections by Shiga toxin-producing Escherichia coli (STEC) in humans have been reported after consumption of mainly beef, but also deer meat. Not only the consumption of contaminated deer meat represents a risk, but also the transmission of STEC between deer and domestic animals should be considered. Within the framework of a telemetry study of roe deer (Capreolus capreolus) the aim was to analyse the occurrence of STEC. Due to the chance to sample some animals several times it was possible to obtain data on the repeated shedding of STEC in roe deer. In total 124 faeces or rectal swabs of 77 live trapped roe deer were collected. The isolates obtained were characterized for stx subtypes, different virulence genes, the so-called top-five serogroups, phylogenetic groups, PFGE-types and antimicrobial susceptibilities. The majority of roe deer were stx-positive whenever sampled. Twenty-eight animals were sampled more than once and were used to examine the duration of shedding STEC. The time interval of 6 persistently stx-negative tested animals was between 6 and 440d (median 49d, interquartile range (IQR) 17-258d). Ten animals excreted undistinguishable STEC strains in intervals between 4 and 778d (median 42d, IQR 22-79d). Most of the isolates were stx2b-positive, eae-negative and frequently ehlyA-positive. None of the isolates belonged to serogroup O26, O103, O111, O145 and O157, respectively. All isolates were sensitive to the antimicrobial substances tested. Although the duration of each shedding event could not be determined the results indicate long-term excretion of STEC in roe deer. This is an important consideration for the observance of good hygiene practice while field dressing of deer and preparing deer meat.

Detection of psychrophilic and psychrotolerant Clostridium spp. in chilled fresh vacuum-packed meat using different PCR methods.

Since 1989, blown pack spoilage has been recognized as a special form of spoilage in vacuum-packed raw and cooked beef. However, only limited information concerning the occurrences of bacteria causing blown pack spoilage on chilled fresh meat is available. In this study, a total of 63 beef and 33 lamb commercially available samples from different countries and without any signs of spoilage were examined for contamination with psychrophilic and psychrotolerant Clostridium spp. using different PCR systems. In total, 34.4% of the chilled fresh vacuum-packed meats were PCR positive. A higher number of lamb samples were identified as PCR positive compared with beef. A geographical relationship between positive results and the origin of the samples could not be determined. PCR system described by Brightwell and Clemens (Development and validation of a real-time PCR assay specific for Clostridium estertheticum and C. estertheticum-like psychrotolerant bacteria. Meat Sci 2012;92:697-703) gave the highest number of positive detections compared with the Broda, Boerema and Bell PCR system (PCR detection of psychrophilic Clostridium spp. causing 'blown pack' spoilage of vacuum-packed chilled meats. J Appl Microbiol 2003;94:515-22). Eight clostridia isolates from two German beef and four Welsh lamb samples were isolated overall. Three of these clostridia isolates were identified as Clostridium estertheticum whereas five clostridia isolates remain unidentified. The study shows that psychrophilic and psychrotolerant Clostridium spp. are more prevalent in retail samples than previously suspected.

Evaluation of a bead-based enzyme immunoassay for the rapid detection of osteocalcin in human serum.

BACKGROUND: Circulating osteocalcin is a well-known marker for bone formation, but none of the commercial kits currently available can be used in automated systems. Here we present the first semiautomated assay for human serum osteocalcin. METHODS: Polystyrene beads were coated with antibodies against the COOH terminus of osteocalcin and used in the COBAS((R)) EIA System. Osteocalcin was detected with peroxidase-conjugated antibodies against the osteocalcin NH(2) terminus. RESULTS: The time required to analyze an unknown sample was 60 min, with a lower detection limit of 4.5 microg/L and a linear dose-response curve between 4.5 and 100 microg/L. The intraassay imprecision (CV) was 5-8% (n = 21); the interassay variation was 6-9% (n = 14). In samples from human volunteers and patients, data generated with the newly developed assay were comparable to those obtained with standard microtiter plate-based assays. CONCLUSIONS: The coated beads assay may be implemented on fully automated analyzers, which not only may further reduce imprecision but may also substantially increase the applicability of osteocalcin as a marker for bone metabolism in the routine clinical setting.

Immunological markers of childhood fevers in an area of intense and perennial malaria transmission.

In order to describe presumed paediatric malaria on a cell-immunological basis, the soluble receptors of IL-2 (sIL-2R) and tumour necrosis factor (sTNF-R55 and sTNF-R75) were quantified in highly exposed young Tanzanian children. Sera were obtained from 66 acute and 72 reported febrile patients during health post consultations and follow-ups and from 68 community controls. Levels of sIL-2R, sTNF-R55 and sTNF-R75 were significantly elevated during fever attacks, especially in very young children. Soluble TNF-R75 levels were most stable and those of sTNF-R55 least. Levels of sTNF-R55 were related to the magnitude of fever and thus appeared to reflect attack severity. Levels of sTNF-R75 were highly significantly associated with parasite density, indicating that this response is malaria-specific. The present study indicates that sTNF-R75 levels could become a useful immunological tool in malaria intervention studies, as they reflect changes in malaria-specific immune responses. Future studies should validate this potential in different endemic settings.

Genital warts do not respond to systemic recombinant interferon alfa-2a treatment during cannabis consumption.

The case of a 22-year-old man suffering from genital warts is described. The lesions responded completely to recombinant interferon alfa-2a only after discontinuation of cannabis consumption. Cannabis was detected using the enzyme immunoassay/1-trans-tetrahydrocannabinoid method in urine. Southern blotting of frozen genital wart biopsy material revealed papillomavirus type 11 DNA, the amount of which increased significantly during interferon treatment. The final clearing of lesions after discontinuation of cannabis consumption implicates that the drug-induced impairment of cellular immunity was reversible. It is concluded that drug abuse and especially cannabis consumption may play some role in the world-wide increase in genital papillomavirus disease and in the high number of recalcitrant courses of genital warts.

Filter spot-ELISA for the enumeration of interferon-alpha antibody-secreting cells.

A filter spot-ELISA was developed for the enumeration of interferon-alpha (IFN-alpha) antibody secretion by murine and human cells. Various human IFN-alpha subtypes were coated onto nitrocellulose membranes by means of broad specificity bovine antibodies. Membranes were blocked with milk proteins, and antibodies released by individual cells during a 3 h culture period were visualized as distinct spots using peroxidase-conjugated, species-specific anti-immunoglobulin antibodies and diaminobenzidine/hydrogen peroxide substrate solution. The spot-ELISA is rapid, easy to perform, and highly sensitive. Possible applications include frequency estimates of IFN-alpha antibody-secreting cells in blood, tissues and hybridoma cultures as well as the evaluation of specificity and immunoglobulin class of the respective antibodies.

Interferon-alpha antibodies in autoimmune diseases.

Circulating autoantibodies constitute a prominent feature of many autoimmune diseases. Recently, antibodies to interferon (IFN) have been recognized as an additional variant occurring spontaneously in about 10% of patients with autoimmune disorders. The reactivity of these antibodies appears to be restricted to IFN-alpha, with antibodies of individual patients recognizing unique epitope patterns. IFN antibodies are preferentially of the IgG type, and they likely constitute part of the normal B-cell repertoire. Whereas antibody titers differ largely among patients, the affinity of IFN binding has not been determined. Likewise, the mechanisms involved in IFN antibody synthesis as well as the clinical impact of these antibodies in autoimmune disorders remain elusive. Based on recent reports and our own observations, this article reviews current knowledge regarding occurrence and properties of IFN antibodies associated with autoimmune conditions. From clinical and experimental evidence, mechanisms possibly involved in IFN antibody production are derived and the significance of antibody-mediated sequelae is evaluated.

Treatment of patients with chronic type B hepatitis and concurrent human immunodeficiency virus infection with a combination of interferon alpha and azidothymidine: a pilot study.

Six patients with chronic type B hepatitis and concurrent infection with the immunodeficiency virus were treated with 600 mg azidothymidine (AZT)/day and 3 X 10(6) units of interferon-alpha (IFN-alpha) every other day for a total of 4 months. None of the patients treated lost the hepatitis B virus (HBV). HBV-DNA concentrations were not significantly influenced by this treatment. Human immunodeficiency virus (HIV) infection was also not affected except for a transient rise in CD 4-positive cells in 2 individuals, who had initially low CD 4-positive cells. Treatment did not influence the presence of HIV-Ag in the serum. In conclusion, a combination therapy of IFN and AZT does not seem to be beneficial at the doses given and the time involved.

Treatment of protracted acute type B hepatitis with recombinant alpha-A-interferon. A pilot study.

Six individuals with protracted acute type B hepatitis were treated with recombinant alpha-A-interferon for 12 weeks. Two females eliminated the HBV completely, while 4 males did not respond. Response was preceded by a flare-up of the liver disease. It appears that responses to interferon are not higher in protracted acute type B hepatitis than in progressed chronic active hepatitis B. This assumption has to be proven in larger studies on a statistical basis.

[Treatment of hepatitis B surface antigen (HBsAg)-positive chronic hepatitis with recombinant alpha-A-interferon. Results of a phase II study]. / Behandlung der Hepatitis-B-Surface-Antigen(HBsAg)-positiven chronischen Hepatitis mit rekombinantem alpha-A-Interferon. Ergebnisse einer Phase-II-Studie.

The HBsAg carrier state may present as chronic active hepatitis which may proceed to cirrhosis of the liver and to primary liver cell carcinoma. The large scale production of interferons made these substances available for long-term treatment. A deficiency in interferon production in chronic type B hepatitis presented the rational to treat this disease with interferon alpha-A. In this phase II-trial 3/31 patients eliminated HBsAg and 14/31 HBeAg. This was followed by normalisation of liver function tests and probably an improved prognosis. Efficiency of treatment was dependent on the interferon dose, the level of viral replication, the level of liver enzymes before treatment and concurrent infections (e. g. HIV infection). Reactivation occurred in five patients suggesting that the treatment period was too short in some individuals. Future studies will potentially improve efficiency by the modification of the interferon schedule and a better understanding of the mode of action of interferon.

[Alpha interferon in the therapy of hairy cell leukemia. Results of 3 prospective multicenter studies in West Germany]. / Interferon alpha für die Therapie der HCL. Ergebnisse von drei Prospektiven multizentrischer Studien in der Bundesrepublik.

Within 19 months of recruitment 51 centers enrolled 84 patients in our prospective multicenter trial for low-dose IFN alpha-2c for HCL (1.2 x 10 IU/m x 28 days s.c.- or lower). Induction therapy of 84 days was followed by maintenance with 2 weekly doses. Patients who reached stage Jansen I or A were then randomized to 2 arms either to receive further IFN or to stop treatment. Reinduction was reinstituted whenever any patient lost Jansen stage A or I. Only 3 patients failed. But enrollment was discontinued when it became apparent that fatalities in splenectomized (SX) patients due to septicemia outnumbered deaths given in the literature. Low-dose IFN administered daily might compromise host defense. Our next study aimed at improving the safety of IFN for SX patients. Pulsatile IFN on 7 consecutive days within a 28-day period was given to all SX patients. A randomization of non-SX patients either to receive daily or pulsatile IFN should provide proof of the efficiency. The intermissions ought to be as suitable for in-vitro investigations as for observations concerning the capacity of IFN to produce 'induction of inducers'. Pulsatile treatment has so far been safe in 13 splenectomized patients. Central diagnostic procedures remained the same during both studies. It seems important to us to use an intermittent schedule for the injection of IFN in SX patients and to restrict splenectomy to selected patients.

Phase II trial of recombinant leukocyte A interferon in advanced malignant melanoma.

A total of 21 patients with advanced metastatic malignant melanoma were treated in this efficacy study of recombinant leukocyte A interferon (interferon alpha-2a). Patients received 18 X 10(6) units interferon alpha-2a by i.m. injection daily for the first 10 weeks and then three times weekly for a further 4 months. The symptoms of toxicity observed in this study resembled those previously reported for alpha interferons and included fever, chills, fatigue, anorexia, myalgia, headache, occasional nausea and vomiting, dose-dependent reversible leukopenia, and hepatic transaminase elevations. Of the 21 patients, 12 had evidence of tumor progression, 6 had stable disease for at least 2 months, and complete remission was seen in 3 patients with stage III melanoma. We conclude that interferon alpha-2a appears to have some antiproliferative effect in metastatic malignant melanoma. While its use in stage IV patients with big tumor masses is doubtful, there seems to be therapeutic benefit in earlier stages.

[Alpha interferon in condylomata acuminata and juvenile diabetes mellitus]. / Interferon-alpha bei Condylomata acuminata und juvenilem Diabetes mellitus.

Persistent condylomata acuminata in a 21-year-old patient with diabetes mellitus were treated with highly purified interferon-alpha (IFN-alpha) obtained by recombinant DNA technology. Daily dose was 1.5 X 10(6) IU, given subcutaneously. Already during treatment the condylomata regressed. Two weeks after the end of therapy, i.e. after a total dose of 10.5 X 10(6) IU IFN-alpha, all condylomata had completely receded. Blood glucose levels remained constant with concomitant insulin therapy. Toxic side-effects or antibodies to IFN-alpha were not observed.

Systemic treatment of condylomata acuminata with recombinant interferon-alpha-2a: low-dose superior to the high-dose regimen.

Fourteen patients with chronic genital warts were entered in a pair-matched crossover trial of recombinant interferon-alpha-2a. They received daily doses of either 1.5 X 10(6) U or 18 X 10(6) U interferon for one week by the subcutaneous route (abdominal skin). After an observation period of 4 weeks in case of no change or partial response, patients of the low-dose group were given 18 X 10(6) U and those of the high-dose group 3 X 10(6) U daily for another 7 days. Eight of 14 patients had a complete remission by the end of the therapy-free interval of 4 weeks, 5 patients of the low-dose group and 3 patients of the high-dose group. Acute side-effects consisted of a transient influenza-like syndrome. Remissions have been maintained for 6 to over 10 months. The data implicate that recombinant interferon-alpha-2a is highly effective in patients with human papillomavirus-related genital warts and that the low-dose regimen is at least equivalent to the high-dose treatment.

[Correlation of the cytochemical classification of acute leukemia in children with the course of their disease (author's transl)]. / Zur Korrelation der zytochemischen Klassifizierung akuter kindlicher Leukämien mit deren klinischem Verlauf.

Acute leukemia in 93 children was cytochemically classified into three groups: 1. the PAS-Typ (acute undifferenciated leukemia) paramyeloblastic leukemia, 2. the AML, and 3. the Acid Phosphatase Typ (SP-Typ). Therapy for the first group differed from that for AML. The acid-Phosphatase-Typ was found in only a few cases, where the acid Phosphatase is good to see in the paranuclear region of the cell. These cases have a bad prognosis. It is proposed to publish even single cases of the acid Phosphatasetype leukemia in order to find the optimal therapy.