Development of an experimental facility for the study of microparticle initiated radio frequency vacuum breakdown.

An ongoing objective in the ion cyclotron range of frequencies (ICRF) systems is the improvement of power coupling to the plasma. During the last decade, this goal has been mainly pursued through the study of the coupling resistance, either by optimizing the antenna layout or by tailoring the scrape-off layer profile with gas puffing. Another approach is to increase the voltage handling capability of the ICRF system, limited by breakdown in the launchers or in the transmission lines. This paper describes the design of the ICRF Breakdown EXperiment (IBEX), a device to investigate fundamental aspects of radio frequency arcs under ICRF-relevant conditions. IBEX can achieve a peak voltage of 48 kV at 54 MHz with a 5 kW input power.

IShTAR: A test facility to study the interaction between RF wave and edge plasmas.

Existence of high electric fields near an RF antenna launcher causes a number of parasitic phenomena, such as arcing and impurity release, which seriously deteriorate the performance of an Ion Cyclotron Range of Frequencies (ICRF) heating scheme in fusion devices. Limited accessibility of the near antenna region in large-scale fusion experiments significantly complicates the associated experimental studies. The IShTAR test facility has been developed with the requirement to provide a better accessibility and diagnosability of plasmas in the direct vicinity of an ICRF antenna. The purpose of this work is to give a detailed description on the experimental setup and the available diagnostics. Furthermore, the paper will demonstrate the capability of the experiment to study phenomena near an ICRF antenna launcher which are relevant for large-scale fusion ion cyclotron resonance heating systems.

[Hepatitis A and E enterically transmitted virus infections of the liver]. / Hepatitis A und E--enteral übertragene Virusinfektionen der Leber.

Hepatitis A virus (a picornavirus) and hepatitis E virus (so far unclassified) are small, non-enveloped and relatively stable RNA viruses with many similar, yet, not identical characteristics. Both viruses are transmitted preferentially by the fecal-oral route. Consequently, their spread is favoured by poor personal hygiene and inappropriate sanitary conditions. Infection can pass subclinically, take an acute and self limiting course, and can also manifest as fulminant hepatitis with liver failure. True chronic disease is unknown. Laboratory diagnosis is preferentially performed by serology, but can also be complemented by assay for viral RNA in stool or serum. Resolution of infection leads to immunity which, in the case of hepatitis A, is known to be fully protective and most likely lifelong. Available hepatitis A vaccines are able to induce a similar state of protection. Vaccines for hepatitis E are under development. Specific antiviral treatment is not yet available, neither for hepatitis A nor for hepatitis E.

[Hepatitis A virus infection. A review]. / Hepatitis A-Virus-Infektion. Ubersicht.

The virus responsible for hepatitis A--hepatitis A virus (HAV)--is a small, spherical, and exceptionally resistant RNA-virus. It is transmitted preferentially by the faecal-oral route and apparently replicates exclusively in the liver. The damage of the liver ensuing from HAV infection most likely does not stem directly from virus replication but is the result of an interaction of cell mediated virus-specific immunity with infected hepatocytes. Infection is usually self limiting, yet, in individual cases may also take a protracted and even relapsing course. True chronic infections, however, are not observed. HAV has a world-wide distribution. In countries where inadequate sanitary conditions prevail, the virus persists in the environment and almost 100% of the population acquires infection in childhood. At that age, infection causes no or only minimal clinical symptoms. Infected individuals nevertheless develop protective, long lasting immunity, probably persisting for entire life. In developed, industrialized countries HAV has ceased to circulate in the environment and the general population. Here, infections predominantly occur in adults travelling to endemic areas or exposed at home to thus infected individuals or members of high risk groups (e.g. children in day care centres, i.v. drug users). With increasing age infections become more and more clinically manifest and at and beyond of adolescence more than 80% of patients develop icteric, in some cases even fulminant and fatal hepatitis. Acute hepatitis A infection can be diagnosed by demonstrating the presence of anti-HAV-IgM antibodies. Immunity following either infection or successful vaccination is assessed by measuring anti-HAV-IgG. Preventive measures rely on strict personal and alimentary hygiene as well as on vaccination with inactivated (killed) hepatitis A vaccines. These vaccines are safe, highly immunogenetic and induce long lasting (> 20 years) protection against hepatitis A. Specific antiviral therapy is not yet available.

[Malaria--chemoprophylaxis 2001]. / Malaria-Chemoprophylaxe 2001.

An estimated 20,000 to 30,000 cases of imported malaria are annually diagnosed in industrialised countries. Some 700 of them concern Swiss travellers and foreign guests. Exposure prophylaxis and chemoprophylaxis for high risk destinations lower the risk of malarial disease. The latter is defined as regular intake of antimalarial drugs in subtherapeutic dosage in order to suppress the development of clinical disease. Drugs are usually taken from one week before travel until four weeks after return from an endemic area. Mefloquine, doxycycline, chloroquine plus proguanil, and presumably soon also atovaquone plus proguanil are available in Switzerland for chemoprophylaxis.

Presence and significance of human parvovirus B19 DNA in synovial membranes and bone marrow from patients with arthritis of unknown origin.

Acute rheumatologic symptoms are frequently associated with human parvovirus B19 (B19) infections. A nested PCR (nPCR) assay was used to test for the presence of parvovirus B19 DNA in synovial fluid and/or synovial membrane specimens obtained from a total of 90 patients with arthritis of unknown origin. Whereas only one out of 73 synovial fluid samples were found positive, 15 (16.7%) out of 90 patients had parvovirus B19 DNA in the synovium. B19 virus DNA was detected in nine bone marrow aspirates subsequently obtained from these 15 patients (60%). Whereas each one of the 15 corresponding blood samples contained anti-B19 IgG antibody, none contained anti-B19 IgM antibody and only one was positive for B19 virus DNA. The blood and synovial fluid samples that contained B19 virus DNA were obtained from the same patient, who also had B19 DNA in synovium and bone marrow. For one patient, two distinct synovial membrane specimens collected 10 months apart tested positive for B19 virus DNA. Parvovirus B19 DNA was also detected in synovial tissue of one out of nine nonarthritic patients serving as control group, who also had anti-B19 IgG circulating antibody. These data illustrate that human parvovirus B19 may persist in bone marrow and synovial tissues of patients with arthritis of unknown origin. In contrast, persistence of B19 virus DNA in synovial fluid is rare. The significance of parvovirus B19 DNA in synovium of healthy patients has to be established.

A nested-PCR assay for the simultaneous amplification of HSV-1, HSV-2, and HCMV genomes in patients with presumed herpetic CNS infections.

To facilitate early diagnosis of herpes virus infection of the central nervous system (CNS), a nested polymerase chain reaction (nPCR) assay was developed to test simultaneously for the presence of HSV-1, HSV-2, and HCMV DNA in the cerebrospinal fluid (CSF) of patients with herpetic CNS disease suspected on clinical grounds. The virus type-specific PCR products were differentiated either by agarose gel electrophoresis or by DNA enzyme immunoassay. Using titrated viral stocks as standards, a sensitivity of at least 0.03 infectious units was obtained for HSV-1, HSV-2 and HCMV and no cross-reactions were recorded. Among 178 CSF specimens (171 patients), which were tested under routine conditions, three contained HSV-1 DNA, one contained HSV-2 DNA and one contained HCMV DNA. No double or triple infection was diagnosed. The presence of inhibitors of the Taq DNA polymerase was monitored by spiking aliquots of the above CSF samples with 3 infectious units each of HSV-1 and HSV-2 or HCMV. Whereas none of 93 samples spiked with HSV-1 and HSV-2 contained inhibitors, the PCR reaction was inhibited in three out of 175 samples (1.7%) spiked with HCMV. The complete procedure which requires only 150 microl of CSF is easily completed within 8 h. Through its speed, reliability and sensitivity, this nPCR assay has met the specific criteria of the diagnostic laboratory.

No evidence for a role of modified live virus vaccines in the emergence of canine parvovirus.

In this study the early evolution and potential origins of canine parvovirus (CPV) were examined. We cloned and sequenced the VP2 capsid protein genes of three German CPV strains isolated in 1979-1980, as well as two feline panleukopenia virus (FPV) vaccine viruses that were previously shown to have some restriction enzyme cleavage sites in common with CPV. Other partial VP2 gene sequences were obtained by amplifying CPV DNA from paraffin-embedded tissues of dogs which were early parvovirus disease cases in Germany in 1978-1979. Sequences were analysed with respect to their evolutionary relationships to other CPV and FPV isolates. Those analyses did not support the hypothesis that CPV emerged as a variant of an FPV vaccine virus. Neither did they reveal ancestral sequences among the very early CPV isolates examined. Other possible sources for the origin of CPV are examined, including the involvement of viruses from wild carnivores.

Evidence for persistence of human parvovirus B19 DNA in bone marrow.

A nested polymerase chain reaction assay (nPCR) was used to investigate the potential of human parvovirus B19 DNA to persist in blood or bone marrow samples obtained either from blood donors or cadaveric bone donors or from patients presenting with clinical signs of parvovirus B19 infection. The presence of parvovirus B19 specific antibody in blood was tested by enzyme immunoassay (EIA). B19 virus genome was not detected in any blood sample of 115 blood donors, of whom 92 (80%) had anti-B19 IgG antibody only as an indication of past infection. In contrast; B19 virus DNA was detected in the bone marrow of 4 out of 45 bone donors. Each one of the serum samples available for 3 of these 4 individuals contained anti-B19 IgG antibody. Among 84 patients with clinical manifestations of parvovirus B19 infection, 17 (20%) had B19 virus DNA in bone marrow. Eight of the latter patients had anti-B19 IgG antibody in their blood but neither anti-B19 IgG nor B19 virus DNA. These data document the ability of parvovirus B19 DNA to persist in the bone marrow of asymptomatic individuals and patients with parvovirus B19 infection suspected on clinical grounds.

[Infectious diseases: current diagnostic methods]. / Infektionskrankheiten: neue diagnostische Methoden.

A microbiological diagnosis is either based on the direct detection of the pathogen or on the reaction of the host (indirect method). Many pathogens, in particular small bacteria and viruses, are undetectable by culture or difficult to cultivate. Molecular methods for the direct detection of such pathogens have been widely introduced in many microbiological laboratories within the past few years. The characteristics of these methods as well as new culture methods and new serological tests are discussed.

Suitability and clinical application of a multiplex nested PCR assay for the diagnosis of herpes simplex virus infections.

A novel multiplex nested polymerase chain reaction (PCR) assay was designed and evaluated for routine diagnosis of herpes simplex virus (HSV) infections in patients with either putative HSV infection of the central nervous system or suspected HSV keratitis. Single-tube amplification of HSV type 1 (HSV-1) or type 2 (HSV-2) DNA extracted from cerebrospinal fluid (CSF) or from keratectomy specimens was followed by differentiation of the virus type-specific PCR products either by agarose gel analysis or by DNA enzyme immunoassay. Among 417 CSF specimens obtained from 395 consecutive patients with clinically suspected HSV infection, 11 (2.6%) were positive for HSV-1 DNA and four (1.0%) probes were positive for HSV-2 DNA. None of the specimens was positive for both HSV-1 and HSV-2 DNA. The genome of HSV-2 was detected in a CSF sample obtained from a woman with meningoencephalitis and genital herpes. The presence of PCR inhibitors was detected in six of 111 (5.4%) reconstructed CSF samples. Inhibition could be removed following extraction with a commercial kit. HSV-1 DNA, but no HSV-2 DNA, was detected in corneal buttons obtained from patients with suspected herpetic keratitis. No contamination has been recorded during the 2-year routine use of this test, which has met the specific requirements of a diagnostic laboratory.

Sequence variability among different parvovirus B19 isolates.

Parvovirus B19 is the causative agent of a variety of clinical manifestations, ranging from asymptomatic to severe infection. The basis for this complex pattern of B19-associated diseases is as yet poorly understood. In general there are two different possibilities: firstly, the infected individuals may have a genetic or acquired predisposition, which renders them susceptible for a certain course of infection; secondly, differences in the B19 genome may result in different outcomes of infection. In order to investigate this second possibility we have partially sequenced the genomes of 20 different B19 isolates derived from serum samples from patients with various B19-associated diseases. Four distinct regions, which cover nearly half of the genome and include parts of the coding regions of all three major B19 proteins-NS1, VP1 and VP2, were selected for sequencing. Comparisons between the different extracted virus isolates at the DNA and protein levels revealed that isolates from patients with persistent parvovirus B19 infection show a tendency towards higher genome variability with respect to isolates derived from persons with acute infection.

RNA-protein interactions at the 3' end of the hepatitis A virus RNA.

The regulative cis-acting terminal RNA structures and the proteins involved in the amplification of the hepatitis A virus (HAV) genome are unknown. By UV cross-linking/label transfer experiments, we have analyzed sequences of the 3'-nontranslated region (3'NTR) and preceding domains of the viral genome for their ability to interact with host proteins. A series of cDNA constructs were used to create genomic- and antigenomic-sense transcripts. The results show that the 3'-NTR-poly(A) interacted with host cell proteins with molecular masses of 38, 45, 57, 84, and 110 kDa only weakly, compared with RNA structures also consisting of 3D-coding regions. Protein p38 was most efficiently labeled after interaction with secondary-structure elements located at the 3' end of the HAV RNA, p38 also interacted with a 5'-terminal RNA probe. Optimal RNA binding was found to be dependent on the salt concentration. The specificity of the RNA-protein interaction was proven by competition assays. These data might indicate that a higher-order structure formed at the junction of the 3Dpol-coding sequence and the 3'-NTR of the HAV genome (putative RNA pseudoknot) significantly improves binding of host proteins and thus suggests that this structure might be essential for the formation of the replication complex initiating minus-strand RNA synthesis.

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C.

The RNA genome of hepatitis A virus (HAV) encodes a giant polyprotein that is putatively cleaved proteolytically into four structural and seven non-structural proteins. So far, most of the proposed non-structural proteins and their respective cleavage sites have not been identified. A vaccinia virus recombinant (vRGORF) containing the complete HAV ORF under the control of the bacteriophage T7 promoter was used to express HAV in recombinant animal cells (BT7-H) that constitutively expressed T7 DNA-dependent RNA polymerase. A HAV-specific 27.5 kDa expression product was identified as peptide 2B. The 27.5 kDa 2B antigen was also found in HAV-infected MRC-5 cells. The N-terminal amino acid residues of the new peptide 2B are Ala-Lys-Ile-Ser-Leu-Phe and polyprotein cleavage between 2A and 2B occurred at amino acids 836-837 (Gln-Ala). Furthermore, heterologous expression in the same system of regions P1-P2 and of the protease 3C (3Cpro) gene, showed that P1-P2 polyprotein is not cleaved autocatalytically but by 3Cpro. Hence, 3Cpro is effective in cleaving the polyprotein 2A-2B junction.

Detection of herpes simplex virus after penetrating keratoplasty by polymerase chain reaction: correlation of clinical and laboratory findings.

BACKGROUND: The study was carried out to investigate the possible correlation of clinical findings, histopathologic features and detection of herpes simplex virus DNA in corneal buttons obtained after penetrating keratoplasty. METHODS: We examined 47 consecutive corneal buttons sent for histopathologic examination by light microscopy and using the polymerase chain reaction for the detection of HSV1 and HSV2. Twenty-one corneal buttons from eyes with bullous keratopathy served as controls. RESULTS: The 47 cases were graded from the clinical information available as unproven, suspected and clinically proven cases of herpetic keratitis. This grading did not correlate to specific histopathologic features or to the results of HSV1 DNA testing. None of the cases were positive for HSV2 DNA. CONCLUSION: HSV DNA was detected in some of the cases of clinically unsuspected herpetic keratitis. This technique of demonstrating the presence or absence of HSV in the cornea after keratoplasty is more reliable than clinical data or histopathologic findings and may be important in cases of recurrent inflammatory episodes involving grafts after keratoplasty.

Association between human parvovirus B19 infection and arthritis.

OBJECTIVE: To gain information concerning the association between parvovirus B19 infection and arthritis. METHODS: Blood or synovial fluid, or both, from a total of 77 adult patients with various arthropathies (rheumatoid arthritis 13; mechanical arthropathies 11; crystal induced arthritis 13; idiopathic mono/oligoarthritis 25; suspicion of viral arthritis 15) were tested for the presence of the viral genome and anti-B19 antibodies. B19 DNA in blood and synovial fluid was investigated by nested polymerase chain reaction, and anti-B19 IgM and IgG antibodies were detected in blood by enzyme immunoassay. RESULTS: A recent parvovirus infection was documented by the presence of anti-B19 IgM antibodies in the blood of 13 patients. B19 DNA, together with anti-B19 IgM and IgG antibodies, were detected in the blood of seven patients who had an acute transient arthritis, putatively of viral origin. Viral DNA was detected in a synovial fluid sample and in the blood of one patient with monoarthritis who had an anti-B19 IgG response only. CONCLUSIONS: The prevalence of anti-B19 IgG antibody in these patients with various forms of arthritis (63%) was within the same range as that in the general population (blood donors). However, for the patients with clinical suspicion of viral arthritis, the increased seroprevalence of anti-B19 IgM and the presence of the B19 genome point to an association between human parvovirus infections and acute forms of arthritis.