OBJECTIVES:To determine the clinical relevance of herpes simplex virus (HSV) viremia episodes in critically ill adult patients. METHODS:1556 blood samples obtained for HSV PCR analysis in Intensive Care Unit (ICU) patients over 4 years were retrospectively analyzed, focusing on the comprehensive analysis of 88 HSV-viremic patients. RESULTS:HSV DNA was detected in 11.8% of samples from the ICU. HSV viral loads remained below 5×10(2) copies/ml in 68.2% of patients and exceeded 10(4) copies/ml in 7.9%. Episodes of HSV-viremia correlated with immunosuppressed status and mechanical ventilation in 79.5% and 65.9% of patients, respectively. Only a subset of patients exhibited HSV-related organ damage, including pneumonia and hepatitis (10.2% and 2.3%, respectively). The mortality rate in HSV-viremic patients was not significantly increased compared to the overall mortality rate in the ICU (27.3% vs. 22.9%, p = 0.33). Only patients with high HSV viral loads tended to have a higher, though non-significant, death rate (57.1%, p = 0.14). CONCLUSIONS:Our results suggest HSV viremia is common in ICU patients, potentially favored by immunocompromised status and mechanical ventilation. The global impact of HSV-viremia on mortality in the ICU was low. Quantifying HSV DNA may help identifying patients at-risk of severe HSV-induced symptoms.

BK virus-associated nephropathy (BKVN) occurs in up to 5% of kidney transplants and is a significant cause of graft loss. Four major subtypes of BKV have been described, with the vast majority of individuals persistently infected with BKV Type I (> 80% of the population). Sequencing of BKV isolates subcloned from BKVN patients revealed a high percentage of variants in the urine (40%) in the VP1 subtyping region. In vitro analysis of several viral variants revealed that all variants recovered from the urine of BKVN patients produced infectious viral particles and were replication competent in cell culture while some of the variants induced cytopathic changes in infected cells when compared to the major BKV subtype, VP1 Type I. These results suggest that rare BKV VP1 variants are more frequently associated with disease and that some variants could be more cytopathic than others in renal transplant recipients.

Background: The NS5A protein of the hepatitis C virus has been shown to be involved in the development of hepatocellular carcinoma.Objectives: In a French multicenter study, we investigated the clinical and epidemiological features of a new HCV genotype lb strain bearing a wide insertion into the V3 domain.Study Design: We studied NS5A gene sequences in 821 French patients infected with genotype 1b HCV.Results: We identified an uncharacterized V3 insertion without ORF disruption in 3.05% of the HCV sequences. The insertion comprised 31 amino-acids for the majority of patients; 3 patients had 27 aminoacids insertions and 1 had a 12 amino-acids insertion. Sequence identity between the 31 amino-acids insertions and the V3 domain ranged from 48 to 96% with E-values above 4e(-5), thus illustrating sequence homology and a partial gene duplication event that to our knowledge has never been reported in HCV. Moreover we showed the presence of the duplication at the time of infection and its persistence at least during 12 years in the entire quasispecies. No association was found with extrahepatic diseases. Conversely, patients with cirrhosis were two times more likely to have HCV with this genetic characteristic (p = 0.04). Moreover, its prevalence increased with liver disease severity (from 3.0% in patients without cirrhosis to 9.4% in patients with both cirrhosis and HCC, p for trend = 0.045).Conclusions: We identified a duplicated V3 domain in the HCV-1b NS5A protein for the first time. The duplication may be associated with unfavorable evolution of liver disease including a possible involvement in liver carcinogenesis. (C) 2015 Elsevier B.V. All rights reserved.

cEVR (p =0.003). We found no relationship with the expression of mir-122 for total cholesterol, triglycerids and HCV viral load. Mir122 expression showed a 50% reduction in HCV infected patients and is not modified at the different stages of fibrosis. Conclusions: Patients IL28B CT/TT who failed to PEG-IFN plus ribavirin presented a reduction of mir-122 expression. Whereas, in vitro mir-122 stimulates HCV replication, there was no correlation between viral load and hepatic mir-122 expression. The stage of the disease was not associated with a modification of mir122 expression. Altogether, these results suggest that the level of expression of mir-122 needs to be maintained to regulate its different activity.

Click here and insert your abstract text. Administration of Markets and Fairs Public Services from Craiova, Iasi, and Timisoara are providing public services consumed by the user, by means of contracts and subscription agreements with individuals or legal entities, either to achieve or to exploit the operation of commercial spaces. Administration is done in order to meet common public interest, but in terms of economic efficiency. Managerial success of the Administration of Markets and Fairs Public Service is determined by its ability to identify customer needs and provide products and/or services to satisfy their need by the attraction of small producers and farmers offering organic products. In public service, the results have to be oriented to citizens concomitantly with cost optimization. This paper presents a comparative analysis of Civil Service for the Administration of Markets and Fairs for three major cities in Romania, with a common set of indicators and indices. The commons variable are: population served, the total population of each city, market areas, their number and their distribution for each city, etc.. This analysis helps halls to get closer to the citizen by identifying directions for action in order to increase the satisfaction of citizens served.

ABSTRACT Respiratory viruses are the leading cause of acute infections in humans. However, the burden of certain respiratory viruses, such as coronaviruses, and the relevance of viral coinfections remain unclear. In this study, we investigated the distribution and seasonal occurrences of respiratory viruses detected by multiplex molecular assay in 6,014 samples from 2008 to 2011 in a French hospital. We assessed the detection frequencies of 14 respiratory viruses and their clinical impact in immunosuppressed and nonimmunosuppressed patients. Furthermore, we explored the preferential association patterns between respiratory viruses in multiple infections. Our results indicated that human rhinovirus/enterovirus (HRV/EV) and coronavirus (HCoV) were frequently detected in respiratory samples (48.81% and 11.74% of infected samples, respectively), and the detection frequencies of these viruses were further increased in immunosuppressed patients. The most common subtypes of HCoV were HCoV-229E (33.80%) and HCoV-HKU1 (32.39%). A sharp increase in the detection frequencies of HCoV-229E and HCoV-HKU1 over several months suggested that these subtypes were epidemic in our population. In immunosuppressed patients, HCoV contributed to upper respiratory tract infections (52%). Evidence did not support lower respiratory tract infections exclusive to a unique HCoV infection. In multiply infected individuals, determined in 6.3% of samples, HRV/EV and HCoV were detected in 33.29% and 22.90% of samples, respectively. Interestingly, nearly 50% of HCoV infections were detected in association with another virus. Since the distributions of respiratory viruses in multiply infected patients were subject to preferential association patterns between viruses, we propose complex interactions between different respiratory viruses and host factors.

POSTERS(0.89±0.52%) versus blood (0.05±0.01%; p = 0.0005) and their numbers correlated with advanced stages of fibrosis (p = 0.022), but not to inflammation.Finally, in vitro supernatants of Tregs induced mRNA up-regulation of profibrogenic markers TIMP1, MMP2, TGF-beta1 and alpha-SMA in resting HSC (p < 0.05 each).Conclusion: Our data demonstrate that in chronic hepatitis C Foxp3 + CD4 + Tregs are enriched in areas of hepatic fibrosis, exhibit up-regulated IL-8 expression and can activate HSC.Thus, in addition to suppression of inflammation adaptive Tregs in hepatitis C play a dual role as regulators of fibrogenesis.

Respiratory tract infections are frequent in young children and are related to viruses in most cases. Multiplex Polymerase chain reaction (PCR) based techniques are valuable tools for describing the spectrum of such viruses. The goal of this study was to assess the correlation of virus detection in samples obtained by nasopharyngeal aspiration and by bronchoalveolar lavage. Both samples were taken at the same time in 30 children with lower respiratory tract infection, and were analyzed by multiplex virus PCR (xTAG (TM) RVP). A strong correlation has been found (P = 0.0002) and the most frequently isolated virus was the entero-rhinovirus spp. These results strengthen the opinion that viruses colonize both the upper and lower respiratory tract. Nasopharyngeal samples should be sufficient to the diagnosis of lower respiratory tract viral infection in immuno-competent children. (C) 2012 Elsevier Masson SAS. All rights reserved.

Objective. - Outbreaks of acute respiratory infections (ARI) are common in institutions for elderly people. We had for objective to investigate clusters of cases (lower respiratory tract infection and influenza-like illness [LRTI/ILI]) in order to improve and validate alert strategies in these institutions.Methodology. - Prospective surveillance for LRTI/ILI was implemented in 11 institutions in Alsace, over five years. Clinical criteria were used to identify infected residents and clusters. Nasopharyngeal swabs were collected and rapid tests (Immunoassay) were performed to identify the influenza virus.Results. - The three week periods were analyzed if three cases or more were recorded during the first week. This analysis demonstrated an important risk of epidemic when this number of cases was reached in healthcare units. The influenza virus (10 clusters) and respiratory syncytial virus ([RSV], two clusters) were identified.Conclusion. - The authors confirmed and emphasized the importance of adequate surveillance for clusters of respiratory tract infection cases. Early identification of an outbreak (three cases) is an important point to prevent transmission, especially during epidemic periods and if a virus is identified in the unit or institution. (C) 2011 Published by Elsevier Masson SAS.

independent groups of patients.Mir-224_1 has been described upregulated in hepato-cellular carcinomas.Moreover, deregulation of miR-217 has been reported in pancreatic cancers.Mir-23a inhibits both the development of B cells and the production of IL-6.Interestingly, in our study mir-122 is slightly up-regulated in NRs compared to SVRs.The levels of expression of miRNAs involved in tumoral processes were higher in the liver of NRs than in SVRs and RRs.Conclusion: NRs and SVRs present different miRNAs hepatic expression, before treatment.In particular, mir-99a, mir-181a-2*, mir-23a and mir-217 were significantly deregulated in NRs compared to SVRs.Thus, treatment response could be predicted with a molecular miRNAs signature.

POSTERSpolymerase inhibitors (NNIs) than for either nucleoside polymerase or for NS3 protease inhibitors.The effect of baseline resistance profile on response to treatment with NNIs is explored here.Methods: A dose-ranging study was conducted in HCV genotype 1infected subjects, consisting of 3 days of NNI monotherapy followed by 12 weeks of NNI in combination with pegIFN a-2a and ribavirin (SOC).ABT-333 was dosed at 400 and 800 mg BID, and ABT-072 was dosed at 100, 300 and 600 mg QD.Results: Rates of cEVR (HCV RNA <25 IU/mL at Week 12) were 75% (12/16) for subjects receiving ABT-333 + SOC and 70% (16/23) for subjects receiving ABT-072 + SOC.Four subjects demonstrated phenotypic resistance (EC 50 > 2 SD above the mean) at baseline; three of the four achieved cEVR.Three additional subjects developed resistance during NNI monotherapy, of which two achieved cEVR.For the three subjects developing resistance after 3 days of monotherapy, HCV RNA levels had decreased >2 log 10 from baseline, suggesting that resistance became detectable following suppression of the susceptible virus population.Among the subjects who failed to achieve cEVR, 10 of 11 carried the IL28 C/T or T/T allele.Conclusions: While ABT-333 and ABT-072 selected resistance in vivo, presence of phenotypic resistance at baseline or emergence during therapy did not affect the likelihood of achieving cEVR when the NNI was combined with SOC.This suggests that SOC and/or the endogenous innate immune response was sufficient to suppress resistant virus in these subjects.Unfavorable IL28 host genotype was associated with failure to achieve cEVR.

Background and Aims:We developed JFH1-based intergenotypic recombinant hepatitis C virus (HCV) cell culture systems, expressing the structural proteins, p7 and NS2 of genotypes 1a, 1b, 2b, 3a, 4a, 5a, 6a and 7a, with growth characteristics comparable to the original 2a(J6/JFH1) culture.Our aim was to further develop this panel to express readily detectable reporter proteins.Methods: Enhanced green fluorescent protein (EGFP) was inserted in domain III of NS5A.After transfection of RNA transcripts or passage of culture supernatant viruses, Huh7.5 cells were examined by visualization of HCV NS5A by flow cytometry (EGFP) and confocal microscopy .The open reading frame sequence of viral genomes amplified from supernatant was directly sequenced.Supernatant HCV RNA and infectivity titers were determined by 5 UTR qPCR and focus forming unit (FFU) assays, respectively.Results: In two independent transfections, J6/JFH1-EGFP spread after an eclipse phase of 13 or 19 days.In contrast, J6/JFH1 spread immediately.After viral passage, J6/JFH1-EGFP showed spread kinetics, RNA and infectivity titers similar to J6/JFH1.Sequencing of viral genomes derived from the two transfection experiments, showed a 40aa and 25aa deletion upstream of the EGFP insert, respectively.In reverse genetic studies, the 40aa deletion conferred cell culture adaptation, since J6/JFH1-EGFPD spread as J6/JFH1 and showed similar peak infectivity titers (~10 4.5 FFU/ml).Interestingly, genotype 1-7 intergenotypic recombinants harboring the identified 40aa deletion and EGFP in NS5A spread as efficiently as J6/JFH1 and showed peak infectivity titers of ~10 4 FFU/ml.The 40aa NS5A deletion also allowed introduction of Renilla Luciferase in NS5A, and it did not affect viability of the original J6/JFH1 recombinant.Conclusions: Viability of a panel of JFH1-based genotype 1-7 reporter viruses with EGFP in domain III of JFH1 NS5A depended on a deletion of a viral NS5A sequence upstream of EGFP.This deletion apparently allows insertion of different reporter genes in NS5A and is not required for viability of the untagged virus.Such reporter viruses will be useful for genotype specific high-throughput fluorescence/luminescence based studies of e.g.HCV receptor interactions or neutralization as well as in vitro and in vivo tracking of HCV NS5A.

Hepatitis C virus (HCV) results in persistent infection in more than 70% of infected individuals despite the development of humoral and cellular immune responses. Following infection, although antibodies targeting epitopes of both structural and non structural proteins are elicited, the virus evades antibody-mediated neutralization. Studies of host neutralizing responses against HCV have been limited by the lack of a convenient tissue culture system for HCV infection. In the past five years in vitro models have been developed to characterize interaction of HCV glycoproteins with host cell entry factors and detect antibodies interfering with HCV entry and infection. These models have been used to characterize targets of neutralizing responses and better understand their impact on the pathogenesis of infection.

Hepatitis C virus (HCV) infection is the main cause of chronic liver disease throughout the world, and may progress to cirrhosis and hepatocellular carcinoma (HCC). Immunological factors, especially cytokines and some host genetic variations, rather than direct HCV action, seem to play an important role in the pathogenesis of HCV infection. Elevated levels of interleukin-18 (IL-18) were described previously for chronically (HCV)infected patients. This study is aimed at investigating IL-18 promoter polymorphisms (-607C/ A and -137G/C) in HCV-infected patients with different disease severities (chronic hepatitis C, liver cirrhosis and HCC) and establishing an association between these polymorphisms and IL-18 plasma concentration with the outcome of chronic HCV infection. The carriage of at least one C allele at position -607 (CC+CA) was associated with a higher risk of cirrhosis and HCC (P=0.032). Compared with controls, HCV-infected patients had significantly higher levels of IL-18 (P=0.0001) that correlate with disease severity (P=0.01, P=0.001, P=0.0006, respectively). In conclusion, we supposed a possible implication of IL-18 promoter polymorphisms in the pathogenesis of chronic HCV infection.

Aim. - Infection with hepatitis C virus (HCV) results in chronic hepatitis in more than 70% of cases. Alterations in the maturation of dendritic cells (DC) might play a rote in the immune system's inability to eliminate the virus, although viral factors that could be involved have not been identified. This study in vitro investigated whether HCV structural proteins affect maturation of monocyte-derived DC.Methods. - HCV proteins (core, El, E2) were expressed by transduction with recombinant adenoviruses of immature DC. The ability of these transduced DC to respond to a maturation stimulus was evaluated by measuring cell surface markers, allogenic lymphocyte stimulation and interteukin (IL)-12 production.Results. - Expression of HCV structural proteins did not modify DC maturation in the presence of lipopotysaccharide, as determined by their phenotype and stimulatory functioning. IL-12 secretion was not affected by HCV protein expression in mature DC.Conclusion. - Our results suggest that HCV structural proteins do not affect maturation of monocyte - derived DC by lipopolysaccharide. These findings are important for further studies to clarify the pathogenesis of chronic HCV infection and towards the rational design of cellular vaccine approaches for immunotherapy against hepatitis C. (C) 2007 Elsevier Masson SAS. All rights reserved.