Association of BK viremia with human leukocyte antigen mismatches and acute rejection, but not with type of calcineurin inhibitor.

INTRODUCTION: BK viremia and polyomavirus-associated nephropathy (PVN) represent a significant problem after kidney transplantation. Both are associated with intensified immunosuppression, but other risk factors and the impact of a screening program on outcome are incompletely understood. METHODS: Here, we report on the short- and long-term outcome of a cohort of patients, who were transplanted in 2006/2007 and included in a newly introduced systematic 3-monthly screening for BK viremia at the University Hospital Zurich. In patients testing positive for BK viremia, screening frequency was intensified and immunosuppression reduced. Patients with suspected PVN underwent transplant biopsy. RESULTS: Among 152 included patients, 49 (32%) tested positive for BK viremia, but only 8 developed biopsy-proven PVN. BK viremia had a significant impact on estimated glomerular filtration rate and proteinuria in the first 2 years. Acute rejection episodes and the number of human leukocyte antigen (HLA) mismatches were the strongest independent predictors of BK viremia in a multiple logistic model. In contrast, no particular immunosuppressive agent or regimen was associated with enhanced risk. CONCLUSION: Taken together, systematic BK viremia screening led to detection of a high percentage of viremic patients. With adjustment of immunosuppression, an excellent outcome was achieved. The independent association of HLA mismatches with BK viremia suggests impaired polyomavirus immunosurveillance in highly mismatched allografts.

Family outbreak of an infection with a recombinant Coxsackie A virus in eastern Switzerland.

PURPOSE: We report on an unusual familial outbreak of a coxsackie virus infection in Switzerland in which five family members were affected. Most of the patients presented with signs of meningitis, and four were hospitalized. METHODS: In three individuals, the virus was detected in the cerebrospinal fluid, pharynx, and stool, respectively. The genome was sequenced in specimens of two patients. RESULTS: The nucleotide sequences of both virus strains were identical. Blast search revealed that the first half of the sequence was 88 % homologous to Enterovirus 75 (EV-75), 87 % with Echovirus 11 (E-11), and 84 % homologous to Coxsackie virus A9 (CV-A9). The second half of the sequence was 77 % homologous to EV-75, 75 % to E-11, and 91 % to CV-A9. CONCLUSION: We propose that the isolated virus strain is a recombinant strain with a 5' untranslated region and with the start of the VP4 sequence originating from E-11/EV-75 and the rest of the genome originating from CV-A9. Interestingly, this novel virus strain showed an exceptional virulence and rapid spread. Two weeks after the initial outbreak in this family, a similar outbreak was observed in a second geographic area roughly 100 km distant to the primary identification site, and another 2 months later this virus strain was found to circulate in the western part of Switzerland some 250 km distant to the primary locus. These findings suggest that genetic recombination has resulted in a novel enterovirus with features of high virulence, contagiosity, and spreading.

Hemophagocytic syndrome caused by primary herpes simplex virus 1 infection: report of a first case.

INTRODUCTION: Hemophagocytic syndrome represents a severe hyperinflammatory condition by activated macrophages. Leading viral triggering agents are Epstein-Barr virus (EBV), cytomegalovirus (CMV), and adenovirus. MATERIALS AND METHODS: We present a patient with Wegener's granulomatosis on azathioprine and prednisone medication, who developed a life-threatening hemophagocytic syndrome. Positive plasma polymerase chain reaction (PCR) with negative serology revealed a primary, disseminated infection with herpes simplex virus-1 as the triggering pathogen. After treatment with acyclovir, high-dose steroids, immunoglobulins, and etoposide, the patient recovered. CONCLUSION: Early diagnosis of potentially underlying infections of hemophagocytic syndrome influences the therapeutic approach. It is important to consider a variety of infectious agents, particularly in immunosuppressed individuals. The reported case emphasizes the importance of screening for herpes simplex virus 1.

Herpes virus and Ménière's disease.

OBJECTIVE: The main goal of this study was to examine the vestibular ganglia from patients with intractable classic Ménière's disease (MD) for the presence or absence of DNA from three neurotropic viruses herpes simplex virus 1 and 2 (HSV1, HSV2) and varicella zoster virus (VZV) and to investigate the hypothesis that MD is associated with virus reactivation within Scarpa's ganglion. STUDY DESIGN: Polymerase chain reaction (PCR) was performed with nested primer sets specific for viral genomic DNA of HSV1, HSV2 and VZV in biopsies of the ganglion scarpae of patients with MD who underwent vestibular neurectomy. Included were patients with MD classified as definite MD according to American Academy of Otolaryngology/Head and Neck Surgery criteria. The ganglion scarpae and ganglion geniculi harvested at autopsy from patients without history of MD or facial palsy served as control specimens. RESULTS: No viral DNA was detected in the vestibular ganglion of 7 patients with definite MD. In 34% of the vestibular ganglia of the control group we detected either HSV1 or VZV. Only one Scarpa's ganglion had both viruses present at the same time. Thirty-two out of 34 ganglia from the geniculate segment of the facial nerve contained either HSV1 and/or VZV genomic DNA. Eight specimens contained both viruses simultaneously. Altogether viral DNA was found in 94% of ganglia. Viral genomic DNA of HSV2 was not detected. CONCLUSION: Although HSV and VZV appear to be present in many ganglion cells throughout the human body, we were unable to find genomic DNA of these viruses in patients with definite MD and disabling vertigo, who underwent vestibular neurectomy. Based on these results, reactivation of HSV1 and VZV in the vestibular ganglion does not seem to play a role in the pathogenesis of MD.

Community-acquired pneumonia in children due to Mycoplasma pneumoniae: diagnostic performance of a seminested 16S rDNA-PCR.

A 16S rDNA-PCR assay for Mycoplasma pneumoniae applied to nasopharyngeal secretion (NPS) or pharyngeal swab (PS) from children with community-acquired pneumonia (CAP) was prospectively compared to serological tests including complement fixation (CF) test, a mu-capture enzyme immuno assay (EIA) for the detection of specific IgM, and an EIA for the detection of specific IgG. During a 24-months-period diagnosis of active M. pneumoniae infection was established in 32 (12.6%) of 253 patients for whom paired sera were available. In the acute phase, the sensitivities of PCR from NPS and PS, CF test, IgM EIA, and IgG EIA were 90.0%, 79.3%, 46.9%, 78.1%, and 59.4%, respectively. The corresponding specificities were 98.1%, 98.6%, 97.6%, 87.1%, and 72.4%, respectively. Thus, the 16S rDNA-PCR assay provides a highly sensitive and accurate tool for the rapid diagnosis of M. pneumoniae infection in children with CAP.

Detection and quantification of cell-free Epstein-Barr virus by polymerase chain reaction and subsequent DNA enzyme immunoassay.

Amplification by polymerase chain reaction and subsequent DNA enzyme immunoassay (DEIA) were employed to determine the number of genome equivalents of cell-free Epstein Barr virus (EBV) DNA in peripheral blood. The assay detected cell-free EBV DNA in the serum of 14 out of 18 patients with primary, productive EBV infection (sensitivity 77.7%) but not in healthy EBV carriers with latent infection (specificity 100%). Our assay has the potential for a clinical diagnostic tool to monitor patients at risk for EBV reactivation and productive infection with subsequent EBV-induced lymphoproliferative diseases.

Rapid detection of Mycoplasma pneumoniae in clinical samples by real-time PCR.

M. pneumoniae is a common causative agent of community-acquired pneumonia in children. The diagnosis of such infections is usually based on serology using complement fixation or, more recently, enzyme-immuno assays. PCR has been shown to be a promising alternative. We have evaluated a real-time PCR assay targeting the P1 adhesion protein gene and compared it to a conventional semi-nested PCR assay with the 16S rDNA as target. Comparison of 147 specimens from 48 patients showed an overall agreement of 97.4%. Real-time PCR proved to be of equal value on clinical specimens as conventional PCR regarding sensitivity and specificity, but is clearly advantageous regarding speed, handling and number of samples that can be analyzed per run.

Improved impression cytology techniques for the immunopathological diagnosis of superficial viral infections.

BACKGROUND: For epidemiological and therapeutic reasons early diagnosis of superficial viral infections is crucial. Conventional microbiological techniques are expensive, time consuming, and not sufficiently sensitive. In this study impression cytology techniques were evaluated to analyse their diagnostic potential in viral infections of the ocular surface. METHOD: A Biopore membrane device instead of the original impression cytology technique was used to allow better quality and handling of the specimens. The impressions were processed, using monoclonal antibodies and immunoperoxidase or immunofluorescence techniques to assess the presence of herpes simplex virus, varicella zoster virus, or adenovirus antigens. Ocular surface specimens from healthy individuals (n = 10) and from patients with suspected viral surface disease (n = 19) were studied. Infected and non-infected cell cultures served as controls. RESULTS: This modified technique of impression cytology allowed the collection of large conjunctival and corneal epithelial cell layers with excellent morphology. Immunocytological staining of these samples provided diagnostic results for all three viruses in patients with viral surface disease. CONCLUSIONS: The use of Biopore membrane devices for the collection of ocular surface epithelia offers new diagnostic possibilities for external eye diseases. Immunopathological methods that are applied directly on these membrane devices can provide virological results within 1-4 hours. This contributes considerably to the clinical management of patients with infectious diseases of the ocular surface.

[The effect of Lactobacillus acidophilus "Solco" on the immunological indices of totally decontaminated mice under complete gnotobiological isolation]. / Vliianie Lactobacillus acidophilus "Solco" na immunologicheskie pokazateli total'no dekontaminirovannykh myshei v usloviiakh obshchei gnotobiologicheskoi izoliatsii.

The possibility of stimulating the immunity of totally decontaminated mice, kept isolated under germ-free conditions, with the use of killed L. acidophilus Solco strains has been studied. As revealed in this study, the oral and intraperitoneal administration of strains O1 and O6 to mice leads to a significant increase in the content of immunoglobulin-synthesizing cells in the jejunal lamina propria of the animals. The oral administration of L. acidophilus Solco strain O6 and the intraperitoneal injection of L. acidophilus Solco strain O1 have been found to lead to a significant rise in the level of luminol-dependent chemiluminescence of mouse peritoneal macrophages. The total decontamination of mice induces the development of secondary immunodeficiency which influences the effectiveness of immunostimulating agents.

[The effect of SolcoTrichovac on the vaginal microflora of patients with a papillomavirus infection associated with a cervical intraepithelial neoplasm]. / Izuchenie vliianiia "Solkotrikhovaka" na vaginal'nuiu mikrofloru u bol'nykh s papillomavirusnoi infektsiei v assotsiatsii s tservikal'noi intraépitelial'noi neoplaziei.

The microbiological study of vaginal secretions of 39 female patients of reproductive age (20-30 years) with papilloma virus infection associated with cervical intraepithelial neoplasia (CIN) was carried out. Of these patients, 28 with papilloma virus infection associated with CINI-II made up group I and II having this infection associated with CINII made up group 2. Dysbiotic disturbances in vaginal bacterial flora, found in these patients, were manifested by a decrease in the isolation rate and number of the lacto- and bifidobacteria simultaneously with the excessive growth of opportunistic bacteria. The results of the oral administration of Solco-Trichovac are indicative of the effectiveness of this preparation, which was confirmed by the data of clinical and bacteriological studies. Together with an increase in the isolation rate of lacto- and bifidobacteria, the level of the contamination of the cervicovaginal niche with opportunistic and pathogenic bacterial strains decreased. The results thus obtained make it possible to recommend Solco-Trichovac for the complex treatment of with papilloma virus infection associated with CIN.

[Povidone-iodine for treatment of adenoviral conjunctivitis: an in vitro study]. / Povidoniod zur Behandlung von Adenoviruskonjunktivitis: eine In-vitro-Studie.

BACKGROUND: Adenoviral conjunctivitis causes high socioeconomic costs due to high contagiousness and therefore the need for extended quarantine. To date the only potentially active, topical antiviral agent is povidone-iodine (PVI). The aim of this study was to investigate the effect of diluted PVI on free adenovirus and adenoviral infected cells as well as to evaluate the cellular toxicity of PVI on non-infected cells. MATERIAL AND METHODS: PVI was diluted to a final concentration of 0.0008 %. Virucidal activity was measured IN VITRO using adenovirus 8 and A549 human epithelial cell cultures. Cytotoxicity effects on healthy cells after short- and long-term exposure to diluted PVI were measured in A549 cell cultures. RESULTS: Exposure to PVI at a concentration of 1:10 (0.8 %) completely extinguishes infectivity of free adenovirus after an exposure time of 10 minutes. PVI is less effective against intracellular adenovirus resulting in a decreased infectivity and viral activity for approximately one day with a narrow spectrum between toxicity and virucidal activity. Healthy epithelial cells can be exposed to PVI for up to 6 hours without a cytotoxic effect. CONCLUSIONS: PVI is highly effective against free adenovirus but less effective against intracellular adenoviral particles in already infected cell. Short- and long-term exposure of PVI causes little cytotoxicity for healthy cells. Therefore, administration of diluted PVI at a concentration of 1:10 is a potential option to reduce contagiousness in cases of adenoviral infections.

Retrospective analysis of varicella zoster virus (VZV) copy DNA numbers in plasma of immunocompetent patients with herpes zoster, of immunocompromised patients with disseminated VZV disease, and of asymptomatic solid organ transplant recipients.

BACKGROUND: Varicella zoster virus (VZV) causes significant morbidity and mortality in immunocompromised patients. Subclinical reactivation has been described in solid organ recipients and has been associated with graft versus host disease in bone marrow transplantation. Newer studies assessing the prevalence and impact of subclinical VZV reactivation in solid organ transplant (SOT) recipients are lacking. METHODS AND RESULTS: In a first step we developed a highly sensitive quantitative polymerase chain reaction (qPCR) assay for VZV DNA with a detection limit of < or = 20 copies/mL. Using this assay, we retrospectively analyzed plasma samples of different patient groups for VZV DNA. VZV DNA was found in 10/10 plasma samples of immunocompetent patients with herpes zoster (VZV copy numbers/mL: mean+/-SEM 1710+/-1018), in 1/1 sample of a human immunodeficiency virus-infected patient with primary VZV disease (15,192 copies/mL) and in 4/4 plasma samples of immunocompromised patients with visceral VZV disease (mean of first value 214,214+/-178,572). All 108 plasma samples of asymptomatic SOT recipients off any antiviral therapy, randomly sampled over 1 year, were negative for VZV DNA. CONCLUSION: Our qPCR assay proved to be highly sensitive (100%) in symptomatic VZV disease. We did not detect subclinical reactivation in asymptomatic SOT recipients during the first post-transplant year. Thus, subclinical VZV reactivation is either a rare event or does not exist. These data need to be confirmed in larger prospective trials.

[Fever and headache]. / Fieber und Kopfschmerzen.

A 30-year-old male patient was admitted to our outpatient clinic because of fever, headache and cerebellar symptoms. Clinically he presented with a slight meningism. After exclusion of a focal intracerebral process by head scanning, a first diagnosis of an aseptic meningitis was made by the analysis of the cerebrospinal fluid. With the hint of repeated tick bites, the diagnosis of an early-summer meningoencephalitis was confirmed. The patient recovered without neurological residua within two months. The clinical course, dates of new epidemiologic studies and problems of vaccination are discussed.

Surveillance of cytomegalovirus after solid-organ transplantation: comparison of pp65 antigenemia assay with a quantitative DNA hybridization assay.

In a multicenter study, 113 blood samples from 19 organ transplant patients were analyzed for cytomegalovirus by the pp65 antigenemia assay and a quantitative DNA hybridization assay. Overall, there was 84% agreement among the results obtained by the two tests. Fifteen of 16 episodes of active infection were detected by both assays. One episode was missed by the pp65 assay, and one patient showed significant DNA-emia but only low-level antigenemia.

Two-centre study comparing DNA preparation and PCR amplification protocols for herpes simplex virus detection in cerebrospinal fluids of patients with suspected herpes simplex encephalitis.

In a two-centre study, the routine DNA preparation and PCR amplification protocols were compared for herpes simplex virus (HSV) detection in cerebrospinal fluids (CSFs) of 43 patients with suspected herpes simplex encephalitis (HSE). The combined clinical, radiological and laboratory results indicated HSE in 6/43 (14%) patients. Discrepant PCR results between the two centres were obtained in 8 (18%) cases consisting of 5 false-positive and 3 false-negative results. Seven out of 8 (88%) discrepant results were associated with the method of CSF preparation using protease K digestion followed by heat inactivation. In contrast, CSF digestion with proteinase K followed by DNA purification on silica spin columns was better yielding discrepant PCR results in only 1 of 78 analyses (1.3%). The results point to the need for standardization and inter-laboratory quality control for routine clinical work.

Intracellular distribution of poliovirus proteins and the induction of virus-specific cytoplasmic structures.

In a susceptible cell, enteroviruses induce a vesiculated region (the "virus-induced vesicles") which is both the site of viral RNA synthesis as well as the site referred to morphologically, as the "cytopathic effect." Proteins of poliovirus (type I, Mahoney) were shown to migrate into the region of the virus-induced vesicles of infected HEp-2 cells. Five proteins (P2-5b, P3-4b, P3-6a, P3-7c, P3-9) were found to be associated with the vesicles themselves, either as intrinsic membrane protein (P3-9) or in a soluble form within the vesicles (P3-4b, P3-7c, and, partially, P3-6a) or bound to a DOC-resistant structure (P2-5b and a small amount of P3-6a). Partial inhibition of the cleavage of the viral polyprotein with ZnCl2 was used to alter the viral protein pattern within the cells. The data obtained indicate that P2-5b is the protein responsible for the formation of the virus-induced vesicles.

[Diagnosis and course of myocarditis: a survey in the medical clinics of Zurich University Hospital 1980 to 1998]. / Diagnose und Verlauf bei Myokarditis: eine Erhebung in den Medizinischen Kliniken des Universitätsspitals Zürich im Zeitraum von 1980 bis 1998.

The clinical picture of myocarditis/myopericarditis is of importance in differential diagnosis, especially in younger patients with suspected myocardial infarction. Myocarditis/myopericarditis commonly presents with chest pain, and the diagnosis is usually established on clinical grounds. However, endomyocardial biopsy is necessary to confirm the diagnosis. We evaluated the characteristics of acute myocarditis over the years 1980-1998 in 54 patients of the Department of Medicine of the University Hospital, Zurich. Two to 6 patients per year were hospitalised with this diagnosis. In most cases the diagnosis was established by a combination of criteria, such as a preceding infection of the upper respiratory tract, thoracic pain, ST segment elevations in different precordial leads followed by T wave inversions, arrhythmias, elevation of cardiac enzymes, reversible hypokinesia by echocardiography and normal coronary arteries. At least 3 of 5 criteria were requested. In a first step we analysed retrospectively all patients with acute myocarditis/myopericarditis in the years 1980-1993. Among 30 cases of acute myocarditis/myopericarditis the following causes could be identified: one influenza B, one Toxoplasma gondii infection, 2 Epstein-Barr infections and one bacterial myocarditis with gram-negative rods. The aetiology of the other 25 cases remained unknown. The majority of myocarditis/myopericarditis healed without complications. One patient with Epstein-Barr myocarditis and one with Toxoplasma gondii infection died. Two patients developed dilated cardiomyopathy. In a second phase we analysed prospectively all cases with acute myocarditis/myopericarditis over the period 1994-1998: 24 patients with acute myocarditis/myopericarditis were hospitalised. At that time coronary angiography and endomyocardial biopsies were performed more frequently. We found 2 patients with giant cell myocarditis and 2 with Toxoplasma gondii infection and HIV, all of whom died. In addition, there were 2 patients with eosinophilic myocarditis, one with Lyme carditis, one with Epstein-Barr myocarditis, one with myopericarditis after Campylobacter enteritis and one histologically proven myocarditis after pneumonia with Haemophilus influenzae. The aetiology of the remaining 13 cases with myocarditis/myopericarditis could not be established. Three patients with probable viral myocarditis developed cardiogenic shock requiring intraaortic balloon pump, and fully recovered. The patient with Lyme carditis manifested with total atrioventricular block and was treated with a temporary pacemaker. One patient with lymphocytic myocarditis required heart transplantation because of terminal heart failure and one female patient with histologically proven diffuse lympho-monocytic myocarditis died of cardiogenic shock. All the other cases healed without complications. Serologies are of little diagnostic value and should be restricted to serologies with therapeutic implications. We believe that the apparent increase in myocarditis/myopericarditis in recent years is a result of better diagnostic tools, such as more specific cardiac enzyme tests, coronary angiography and endomyocardial biopsies. In most cases the therapy remains symptomatic. In elected, severe cases steroids and other immunosuppressive drugs are sometimes used.

Effect of UV irradiation on the expression of vaccinia virus gene products synthesized in a cell-free system coupling transcription and translation.

The effect of UV irradiation on the expression of the vaccinia virus genome was investigated in a cell-free system coupling transcription with translation. Exposure of vaccinia virus to an increasing dose of irradiation resulted in differential reduction in the syntheses of virus-specified polypeptides in the coupled system, with sensitivity being proportional to the size of the gene pro duct. This suggests that each translationally functional mRNA species produced in vitro by vaccinia virus cores is synthesized from an individual promoter site.

Accumulation of poliovirus proteins in uninfected isolated HEp-2 cell nuclei in vitro.

Poliovirus (type 1, Mahoney) proteins preferentially inhibited in vitro RNA polymerase II activity in uninfected isolated HEp-2 cell nuclei, as demonstrated by electron microscopic autoradiography. Structural integrity of the nuclei and preincubation of nuclei and poliovirus proteins in vitro prior to addition of [3H]-UTP were prerequisites for differential inhibition. During in vitro incubation, [3H]-labeled poliovirus proteins were shown to accumulate in the uninfected isolated nuclei. Similar poliovirus proteins accumulated in the nuclei to higher relative amounts as compared to the outside or were excluded from the nuclei after in vitro incubation, as in intact poliovirus-infected cells. Since the in vitro transcription system of isolated nuclei exhibited two characteristics of poliovirus infection, i.e., preferential inhibition of RNA polymerase II activity and accumulation of poliovirus proteins in the nuclei, it may be useful to study host RNA synthesis inhibition.