Genetic diversity and occult hepatitis B infection in Africa: A comprehensive review

BACKGROUND Occult hepatitis B infection(OBI)is a globally prevalent infection,with its frequency being influenced by the prevalence of hepatitis B virus(HBV)infection in a particular geographic region,including Africa.OBI can be transmitted th-rough blood transfusions and organ transplants and has been linked to the development of hepatocellular carcinoma(HCC).The associated HBV genotype influences the infection.AIM To highlight the genetic diversity and prevalence of OBI in Africa.METHODS This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and involved a comprehensive search on PubMed,Google Scholar,Science Direct,and African Journals Online for published studies on the prevalence and genetic diversity of OBI in Africa.RESULTS The synthesis included 83 articles,revealing that the prevalence of OBI varied between countries and population groups,with the highest prevalence being 90.9%in patients with hepatitis C virus infection and 38%in blood donors,indicating an increased risk of HBV transmission through blood transfusions.Cases of OBI reactivation have been reported following chemotherapy.Genotype D is the predominant,followed by genotypes A and E.CONCLUSION This review highlights the prevalence of OBI in Africa,which varies across countries and population groups.The study also demonstrates that genotype D is the most prevalent.

Recombinant adenovirus containing hyper-interleukin-6 and hepatocyte growth factor ameliorates acute-on-chronic liver failure in rats

AIM: To investigate the protective efficacy of recombinant adenovirus containing hyper-interleukin-6(HyperIL-6, HIL-6) and hepatocyte growth factor(HGF)(AdHGF-HIL-6) compared to that of recombinant adenovirus containing either HIL-6 or HGF(Ad-HIL-6 or Ad-HGF) in rats with acute-on-chronic liver failure(ACLF).METHODS: The recombinant adenoviruses containing HIL-6 and/or HGF were constructed. We established an ACLF model, and rats were randomly assigned to control, model, Ad-GFP, Ad-HIL-6, Ad-HGF or AdHGF-HIL-6 group. We collected serum and liver tissue samples to test pathological changes, biochemical indexes and molecular biological indexes.RESULTS: Attenuated alanine aminotransferase, prothrombin time, high-mobility group box 1(HMGB1), endotoxin, tumour necrosis factor(TNF)-α and interferon-γ were observed in the Ad-HGF-, Ad-HIL-6- and Ad-HGFHIL-6-treated rats with ACLF. Likewise, reduced hepaticdamage and apoptotic activity, as well as reduced HMGB1 and Bax proteins, but raised expression of Ki67 and Bcl-2 proteins and Bcl-2/Bax ratio were also observed in the Ad-HGF-, Ad-HIL-6- and Ad-HGF-HIL-6-treated rats with ACLF. More significant changes were observed in the Ad-HGF-HIL-6 treatment group without obvious side effects. Furthermore, caspase-3 at the protein level decreased in the Ad-HIL-6 and Ad-HGFHIL-6 treatment groups, more predominantly in the latter group.CONCLUSION: This study identifies that the protective efficacy of Ad-HGF-HIL-6 is more potent than that of Ad-HGF or Ad-HIL-6 in ACLF rats, with no significant side effects.

Role of the HLA-DQ locus in the development of chronic gastritis and gastric carcinoma in Mexican patients

AIM: To determine the HLA-DQ locus in Mexican patients with Chronic gastritis and gastric adenocarcinoma. METHODS: Oligotyping for HLA-DQ locus was performed in 45 Mexican patients with chronic gastritis and 13 Mexican patients with diffuse-type gastric adenocarcinoma, and was then compared with 99 clinically healthy unrelated individuals. H pylori infection and CagA status were assessed in patients by enzyme-linked immunosor-bent assay (ELISA) method. RESULTS: We found a significant increased frequency of HLA-DQB1*0401 allele in H pylori-positive patients with chronic gastritis when compared with healthy subjects [19 vs 0%, P= 1 × 10-7, odds ratio (OR) = 4.96; 95% confidence interval (95% CI), 3.87-6.35]. We also found a significant increased frequency of HLA-DQB1*0501 in patients with diffuse-type gastric carcinoma in comparison with healthy individuals (P = 1 × 10-6, OR = 13.07; 95% CI, 2.82-85.14). CONCLUSION: HLA-DQ locus may play a different role in the development of H pylori-related chronic gastritis and diffuse-type gastric adenocarcinoma in the Mexican Mestizo population.

Cultural Isolation and Characteristics of the Blood Microbiome of Healthy Individuals

Background: On the analogy of the non-pathogenic microbiota found in oral cavity, skin and gastrointestinal tract, existence of blood microbiota was confirmed by DNA sequencing, but never deeply characterized. Hypothesis for the existence of dormant blood microbiota in healthy humans have been arisen and single species have been isolated. The aim of our study was to resuscitate and investigate the biodiversity of bacterial and fungal dormant blood microbiota in healthy individuals by blood culturing and NGS DNA sequencing. Results: Twenty eight blood samples of healthy individuals, seven for each blood type, were studied. Several culture media were tested. Blood microbiota resuscitation was performed in BHI broth supplemented with vitamin K 1 mg/ml, 2% sucrose, 0.25% sodium citrate and 0.2% yeastolate at 43?C for 72 h. All tested blood samples were culture positive, as confirmed by Gram staining and TEM. TEM images demonstrated well defined cell structures. Analysis for bacterial and eukaryotic species was performed by 16S rRNA and ITS2 targeted sequencing. The obtained sequences were clustered (≥97% identity) in Operational Taxonomic Units (OTUs). Among cultured and uncultured samples we identified OTUs similarity with 47 bacterial orders belonging to 15 phyla and 39 fungi orders blonging to 2 phyla. For the first time we demonstrated isolation and sequencing identification of fungal blood microbiota in healthy individuals. Blood-group differences were identified among the bacterial microbiome compositions. Conclusion: The dormant blood microbiome is innate of the healthy individuals. Interventional strategies to bind the host blood microbiome with the states of health and disease remain an unmet research goal.

Chronic atrophic endometritis and pyometra in a ferret: A case report

The aim of this report was to describe a clinical case of chronic atrophic endometritis as a complication of cystic endometrial hyperplasia-pyometra complex in a non-spayed ferret. The ferret was presented with a slight abdominal distension and odorless purulent vulvar discharge after unsuccessful medical treatment with enrofloxacine and aglepristone 2 months ago in another clinic. Ultrasonography revealed enlarged uterine horns filled with fluid and blood laboratory analysis showed anaemia and leukocytosis, so diagnosis of pyometra was made. Laparotomy and ovariohysterectomy were performed. Histopathological and microbiological examination of the uterus revealed the presence of purulent atrophic endometritis caused by Staphylococcus spp. In conclusion, this is a very rare case of endometrial atrophia after chronic uterine inflammation in a ferret.

Mayaro virus infection, the next epidemic wave after Zika?Evolutionary and structural analysis

Objective: To evaluate the evolution of the pathogen Mayaro virus, causing Mayaro fever(a mosquito-borne disease) and to perform selective pressure analysis and homology modelling.Methods: Nine different datasets were built, one for each protein(from protein C to non-structural protein 4) and the last one for the complete genome. Selective pressure and homology modelling analyses were applied. Results: Two main clades(A and B) were pointed in the maximum likelihood tree. The clade A included five Brazilian sequences sampled from 1955 to 2015. The Brazilian sequence sampled in 2014 significantly clustered with the Haitian sequence sampled in 2015. The clade B included the remaining 27 sequences sampled in the Central and Southern America from 1957 to 2013. Selective pressure analysis revealed several sites under episodic diversifying selection in envelope surface glycoprotein El, non-structural protein 1 and nonstructural protein 3 with a posterior probability P≤0.01. Homology modelling showed different sites modified by selective pressure and some protein-protein interaction sites at high interaction propensity. Conclusion: Maximum likelihood analysis confirmed the Mayaro virus previous circulation in Haiti and the successful spread to the Caribbean and USA. Selective pressure analysis revealed a strong presence of negatively selected sites, suggesting a probable purging of deleterious polymorphisms in functional genes. Homology model showed the position 31, under selective pressure, located in the edge of the ADP-ribose binding site predicting to possess a high potential of protein-protein interaction and suggesting the possible chance for a protective vaccine,thus preventing Mayaro virus urbanization as with Chikungunya virus.

Quantification of the Expression of HIF-1alpha by Real-Time PCR in Rat Hepatocytes Cultures Invaded by <i>Shigella flexneri</i>under Normoxic and Hypoxic Conditions

Aim: Shigella flexneri (S. flexneri) is a gram-negative enterobacterium responsible for severe intestinal end systemic infection in humans. The bacteria can reach the liver due to degeneration of the colonic epithelium. Hypoxia is present in many human diseases and can induce the expression of the transcription factor HIF-1alpha that may have a cell protective role. The influence of hypoxia and HIF-1alpha on bacterial infection, studied in this work, is unclear. Hypoxia inducible factor-1alpha (HIF-1alpha) is a transcription factor that acts as a master regulator of gene expression induced by hypoxia. Methods: We compared the ability of S. flexneri to invade rat hepatocytes in primary culture both in normoxic and hypoxic conditions. We evaluated TNF-alpha released by hepatocytes, apoptosis rate and HIF-1alpha expression by confocal microscopy as well as real time PCR technique. Results: We showed that S. flexneri invaded less hepatocytes previously submitted to 24 h hypoxia (6.5% O2) than those cultivated in normoxia (21% O2). S. flexneri also induced HIF-1α expression in hepatocytes, TNF-α secretion and apoptosis. Conclusion: a) Hypoxia alone was not a stimulus to TNF-α secretion, but induced cell apoptosis and HIF-1α expression;b) S. flexneri was able to invade rat hepatocytes and hypoxia apparently influenced significantly bacterial cell invasiveness;c) HIF-1α was expressed in hypoxic conditions, and it was also stimulated by S. flexneri.

Deficiency of Vitamin D in HIV Infected Patients and Its Effect on Some of the Immunological Parameters

Today the HIV infection is a chronic disease with significantly longer duration of the life of the patients. Problems of pressing interest are the persistent immune activation and chronic inflammation during the treatment with antiretroviral therapy. Taking this into account, different factors which could affect the immune system and the progress of the HIV infection are being researched. Vitamin D (25(OH)D) is one of those factors if we take note of its effect on the innate and acquired immunity. The aim of this study was to assess 25-hydroxyvitamin D (vitamin D) status in one part of the Bulgarian HIV-infected adult population and to assess connection between 25-hydroxyvitamin D (vitamin D) status and plasma levels of some major cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ). The study includes 145 HIV-positive patients, who are being monitored in the Department for acquired immune deficiency at Specialized Hospital for Infectious and Parasitic Diseases “Proff. Ivan Kirov”—Sofia. From all of the monitored patients only in 15% of the tested we found normal 25(OH)D serum levels, and in 12% of the patients we found deficiency. The largest group is that of patients with insufficiency of vitamin D. We didn’t discovered significant difference in the 25(OH)D average values between men and women. There were no significant differences in the average values of the 25(OH)D serum levels when dividing the patients according to their antiretroviral therapy, but after separating the patients by gender, we found that the untreated women had average values of 25(OH)D higher than that of the women treated with EFV. On the next stage of the survey on the 60 HIV-infected patients, who are from the first tested group, we additionally defined the cytokine profile. Our results suggests that increasing 25(OH)D deficiency worsens the damaging of the cellular immune response. The lower levels of vitamin Dare associated with increased levels of IL-6, decreased levels of IL-10, IFN-γ and TNF-α. There’s active immune inflammation when there are reduced 25(OH)D serum levels and it leads to stimulated secretion of the regulatory cytokines and suppression of the Th1 antiviral response. The phase of advanced 25(OH)D deficiency is characterized by parallel depletion of the regulatory and effecter capabilities of CD4 lymphocytes. The recovery of the CD4 lymphocyte pool is difficult because of the lower than average 25(OH)D serum levels, regardless of the conducted antiretroviral therapy.

Evaluation of a Live Paratuberculosis Vaccine in Endemically Infected Flocks of Sheep and Goats in Greece

A large 7-year vaccination trial was conducted in 15 flocks of goats and 7 flocks of sheep, known to be infected with Mycobacterium avium subsp. paratuberculosis (MAP), in Northern Greece. A total of 3665 kids and 1685 lambs, 7 - 30 days old, were vaccinated during 1995-1999. Seven hundred and seventy-five kids and 413 lambs were kept as unvaccinated controls. For each trial, the Incidence Rate Ratio (IRR), with respective exact 95% confidence intervals, was calculated. All IRR point estimates for young animals were very large (from 5.68 to 11.78 for kids and from 4.28 to 10.08 for lambs), while none of the 95% confidence intervals included 1. The protective effect of vaccination was large and the difference in mortality among vaccinated and unvaccinated animals was more pronounced in young animals. The effect in adult animals was smaller than in young animals;it was, however, still considerable. Upon visual inspection of the K-M curves, it seems that for the young animal trials the vaccinated and control-group curves were diverging increasingly over time, which indicates that the gain from the vaccination (or the loss from non-vaccination) might increase over time during the trial.

Previous Pulmonary Fibrosis in Dermatomyositis/Polymyositis: A Predictive Factor for Pulmonary and Extra-Pulmonary Tuberculosis

Objective: With scant studies in the literature, little is known about the risk factors for tuberculosis in patients with dermatomyositis/polymyositis. Therefore, the aim of the present study was to analyze the predictive factors for tuberculosis development in dermatomyositis/polymyositis. Methods: This single-center, retrospective, cohort study initially included 290 patients with dermatomyositis/polymyositis, from 2002 to 2016. Tuberculosis (pulmonary and/or extra-pulmonary) was confirmed after dermatomyositis/polymyositis diagnosis in 12 patients (4.1%) (Tuberculosis+ group). For the control group (Tuberculosis&#8722;), 24 patients without tuberculosis were arbitrarily selected in the same period and matched for age, ethnicity, gender, age at disease diagnosis, disease duration and type (dermatomyositis or polymyositis). Results: Tuberculosis occurred for a median of 16 months after dermatomyositis/polymyositis diagnosis. Clinical, laboratory and treatment features were similar in Tuberculosis+ and Tuberculosis&#8722;groups (P > 0.05). However, previous pulmonary fibrosis in dermatomyositis/polymyositis was more prevalent in the Tuberculosis+ group (41.7 vs. 8.3%;P = 0.029). Moreover, on a multivariate logistic regression model, pulmonary fibrosis was significantly associated with Tuberculosis (Odds ratio: 9.59, 95% confidence interval: 1.17 - 78.82). Tuberculosis affected 3 dermatomyositis cases for every 1 polymyositis case, with predominantly pulmonary followed by extra-pulmonary involvement (pleura, cutaneous, muscular, joint, soft tissue and hematologic). Two or more sites were affected in 41.7% of cases. Conclusions: Previous pulmonary fibrosis in dermatomyositis/polymyositis was a predictive factor associated with tuberculosis development. Further studies are needed to confirm these results.

Chronic Epstein-Barr virus-related hepatitis in immunocompetent patients

AIM: To investigate reactivated Epstein-Barr virus (EBV) infection as a cause for chronic hepatitis. METHODS: Patients with occasionally established elevated serum aminotransferases were studied. HIV, HBV and HCV-infections were excluded as well as any other immunosuppressive factors, metabolic or toxic disorders. EBV viral capsid antigen (VCA) IgG and IgM, EA-R and EA-D IgG and Epstein-Barr nuclear antigen (EBNA) were measured using IFA kits. Immunophenotyping of whole blood was performed by multicolor flow cytometry. CD8+ T cell responses to EBV and PHA were determined according to the intracellular expression of IFN-γ. RESULTS: The mean alanine aminotransferase (ALT) and gamma glutamyl transpeptidase (GGTP) values exceeded twice the upper normal limit, AST/ALT ratio < 1. Serology tests showed reactivated EBV infection in all patients. Absolute number and percentages of T, B and NK cells were within the reference ranges. Fine subset analysis, in comparison to EBV+ healthy carriers, revealed a significant decrease of naive T cells (P < 0.001), accompanied by increased percentage of CD45RA- (P < 0.0001), and terminally differentiated CD28-CD27- CD8+ T cells (P < 0.01). Moderately elevated numbers of CD38 molecules on CD8+ T cells (P < 0.05) proposed a low viral burden. A significantly increased percentage of CD8+ T cells expressing IFN-γ in response to EBV and PHA stimulation was registered in patients, as compared to controls (P < 0.05). Liver biopsy specimens from 5 patients revealed nonspecific features of low-grade hepatitis.CONCLUSION: Chronic hepatitis might be a manifestation of chronic EBV infection in the lack of detectable immune deficiency; the expansion of CD28- CD27- and increase of functional EBV-specific CD8+ T cells being the only surrogate markers of viral activity.

Hepatitis C virus-related B cell subtypes in non Hodgkin's lymphoma

AIM:To evaluate if indolent B cell-non Hodgkin's lymphoma(B-NHL) and diffuse large B-cell lymphoma(DLBCL) in hepatitis C virus(HCV) positive patients could have different biological and clinical characteristics requiring different management strategies.METHODS:A group of 24 HCV related B-NHL patients(11 indolent,13 DLBCL) in whom the biological and clinical characteristics were described and confronted.Patients with DLBCL were managed with the standard of care of treatment.Patients with indolent HCV-related B-NHL were managed with antiviral treatment pegylated interferon plus ribavirin and their course observed.The outcomes of the different approaches were compared.RESULTS:Patients with DLBCL had a shorter duration of HCV infection and a higher prevalence of HCV genotype 1 compared to patients with indolent B-NHL in which HCV genotype 2 was the more frequent genotype.Five of the 9 patients with indolent HCV-relatedB-NHL treated with only antiviral therapy,achieved a complete response of their onco-haematological disease(55%).Seven of the 13 DLBCL patients treated with immunochemotheraphy obtained a complete response(54%).CONCLUSION:HCV genotypes and duration of HCV infection differed between B-NHL subtypes.Indolent lymphomas can be managed with antiviral treatment,while DLBCL is not affected by the HCV infection.

Immune thrombocytopenic purpura induced by intestinal tuberculosis in a liver transplant recipient

A variety of clinical manifestations are associated directly or indirectly with tuberculosis. Among them, haematological abnormalities can be found in both the pulmonary and extrapulmonary forms of the disease. We report a case of immune thrombocytopenic purpura(ITP) associated with intestinal tuberculosis in a liver transplant recipient. The initial management of thrombocytopenia, with steroids and intravenous immunoglobulin, was not successful, and the lack oftuberculosis symptoms hampered a proper diagnostic evaluation. After the diagnosis of intestinal tuberculosis and the initiation of specific treatment, a progressive increase in the platelet count was observed. The mechanism of ITP associated with tuberculosis has not yet been well elucidated, but this condition should be considered in cases of ITP that are unresponsive to steroids and intravenous immunoglobulin, especially in immunocompromised patients and those from endemic areas.

Interleukin-6-174G/C polymorphism is associated with a decreased risk of type 2 diabetes in patients with chronic hepatitis C virus

BACKGROUND Chronic hepatitis C(CHC)is associated with type 2 diabetes mellitus.Although the pathogenesis remains to be elucidated,a growing evidence has suggested a role of pro-inflammatory immune response.Increased serum concentrations of Interleukin 6(IL-6)have been associated with insulin resistance,type 2 diabetes mellitus as well as advanced forms of liver disease in chronic hepatitis C infection.AIM To investigate the frequency of IL-6-174G/C(rs1800795)single nucleotide polymorphism(SNP)in CHC patients and in healthy subjects of the same ethnicity.Associations between type 2 diabetes mellitus(dependent variable)and demographic,clinical,nutritional,virological and,IL-6 genotyping data were also investigated in CHC patients.METHODS Two hundred and forty-five patients with CHC and 179 healthy control subjects(blood donors)were prospectively included.Type 2 diabetes mellitus was diagnosed according to the criteria of the American Diabetes Association.Clinical,biochemical,histological and radiological methods were used for the diagnosis of the liver disease.IL-6 polymorphism was evaluated by Taqman SNP genotyping assay.The data were analysed by logistic regression models.RESULTS Type 2 diabetes mellitus,blood hypertension and liver cirrhosis were observed in 20.8%(51/245),40.0%(98/245)and 38.4%(94/245)of the patients,respectively.The frequency of the studied IL-6 SNP did not differ between the CHC patients and controls(P=0.81)and all alleles were in Hardy-Weinberg equilibrium(P=0.38).In the multivariate analysis,type 2 diabetes mellitus was inversely associated with GC and CC genotypes of IL-6-174(OR=0.42;95%CI=0.22-0.78;P=0.006)and positively associated with blood hypertension(OR=5.56;95%CI=2.79-11.09;P<0.001).CONCLUSION This study was the first to show that GC and CC genotypes of IL-6-174 SNP are associated with a decreased risk of type 2 diabetes mellitus in patients chronically infected with hepatitis C virus.The identification of potential inflammatory mediators involved in the crosstalk between hepatitis C virus and the axis pancreas-liver remains important issues that deserve further investigations.

A New Approach to Reducing Mortality from Dengue

In 2009, based on a multicenter study conducted in Asia and Latin Americaand subsidized by the Dengue Control (DENCO) Research Program, the World Health Organization (WHO) proposed a new classification for dengue cases. The purpose of the present study was to evaluate the applicability of the new classification, relative to its previous version [1]. The evaluation, conducted in Campo Grande county, Mato Grosso do Sul state, Brazil, drew on secondary data from referral healthcare centers that assist high-severity dengue patients. A total of 156 medical records of patients with laboratory diagnosis of dengue were investigated. The records covered two epidemic periods: summer of 2006-2007 and summer of 2009-2010. The results showed that 64.6% of cases classified as dengue fever under the 1997 criteria presented manifestations of severity, warranting their reclassification as dengue with warning signs (49) or severe dengue (15) under the 2009 revised criteria. Bleeding, persistent vomiting, and severe, continuous abdominal pain were the most prevalent warning signs, indicative of risk of progression to severe disease. The revised classification was proved less complex than the current version, facilitating the identification of cases and the clinical management of patients.

Diversity of Human Immunodeficiency Virus Type-1 Subtypes in Western Kenya

Background: HIV/AIDS is the principal pandemic in the world today.?Two viral types (HIV-1 and HIV-2), with numerous groups (M, N and O for HIV-1 and A through H for HIV-2) have emerged. These have further proliferated into numerous subtypes, sub-subtypes and circulating recombinant forms (CRF) over the last 30 years.?HIV-1 variants circulate together within a geographical region providing an opportunity for recombination of viral strains within infected individuals.?In Kenya, at least nine different genetic HIV-1 subtypes and several recombinant forms have been defined within group M, which accounts for the majority of cases in the AIDS pandemic. Objective:?To determine the genetic diversity of HIV-1 in the western region of Kenya bordering Uganda. Methodology: A cross sectional study was carried out at Busia District Hospital between 2007 and 2009. A total of 75 patients were sampled randomly from a cohort of 1000 clients on antiretroviral therapy. Blood samples were analysed at the HIV Laboratory, Kenya Medical Research Institute, Nairobi, Kenya. PCR was carried out on the?Pol?region of HIV, sequenced and analysed by BLAST for subtypes. Results: BLAST analysis revealed the following circulating subtypes: 40/75 (53.30%) were HIV-1 group M subtype A1;21/75 (28.0%) were subtype D;5/75 (6.7%) were subtype G;4/75 (5.30%) were subtype C;and 2/75 (2.70%) were subtype A2. Only one isolate was identified for the other subtypes?viz: 1/75 (1.30%) resembled subtype B;1/75 (1.30%) was A1/C, and 1/75 (1.30%) was A1/D. Conclusion: The study showed increasing HIV-1 diversity along the Kenya-Uganda border with the emergence of A1/C and A1/D recombinants. Such HIV-1?diversityvis a vis?the recent expanded access to antiretroviral therapy in resource limited settings calls for continuous evaluation of anti-HIV regimens. There?is need therefore, for regular surveillance and monitoring for mutations that are likely to lead to drug resistance if we have to achieve successful treatment outcomes.

Viral infections and implications for male reproductive health

Viral infections have haunted humankind since times immemorial.Overpopulation,globalization,and extensive deforestation have created an ideal environment for a viral spread with unknown and multiple shedding routes.Many viruses can infect the male reproductive tract,with potential adverse consequences to male reproductive health,including infertility and cancer.Moreover,some genital tract viral infections can be sexually transmitted,potentially impacting the resulting offspring's health.We have summarized the evidence concerning the presence and adverse effects of the relevant viruses on the reproductive tract(mumps virus,human immunodeficiency virus,herpes virus,human papillomavirus,hepatitis B and C viruses,Ebola virus,Zika virus,influenza virus,and coronaviruses),their routes of infection,target organs and cells,prevalence and pattern of virus shedding in semen,as well as diagnosis/testing and treatment strategies.The pathophysiological understanding in the male genital tract is essential to assess its clinical impact on male reproductive health and guide future research.

Granulicatella adiacens Infective Endocarditis Treated With Teicoplanin and Ceftriaxone:A Case Report

We describe a case of infective endocarditis(IE)caused by Granulicatella adiacens in a patient who had fever for 4months before admission.He had a complex medical history of active hyperthyroidism and schizophrenia,which might have misled the clinician in ignoring the existence of a serious infection like IE.A history of splenectomy and the likelihood of congenital cardiac valve deficiency were the possible underlying risk factors.His persistent leukocytosis perplexed our judgement in terms of resolution of infection and treatment efficacy,which was probably due to his splenectomy.C-reactive protein was the alternative laboratory indicator.Clinical and Laboratory Standards Institute criteria have not been established for the fastidious G.adiacens.Therefore,we referred to the viridans group streptococci and literature to evaluate antimicrobial susceptibility testing.We suggest that teicoplanin combined with ceftriaxone could serve as an effective therapy for IE caused by G.adiacens.

Role of TMS5： staphylococcal multidrug-efflux protein QacA

Background QacA, a main exporter mediating the multidrug-resistance of Staphylococcus aureus to a vanety of antiseptics and disinfectants, possesses a topology of 14 a-helical transmembrane segments （TMS）. Our study aimed to determine the importance and topology of amino acid residues in and flanking the cytoplasmic end of TMS5. Methods Site-directed mutagenesis was used to mutate 5 residues, including L146, A147, V148, W149 and S150, into cysteine. A minimum inhibitory concentration （MIC） and transport assay with or without N-ethylmaleimide （NEM） were performed to analyse the function of these mutants. Results All of the mutants showed comparable protein expression levels. MIC analysis suggested that mutant W149C showed low resistance levels to the drugs, but the mutations at L146, A147, V148, and S150C had little or no effect on the resistance level. And the results of the fluorimetdc transport assay were in agreement with those of MIC analysis, that is to say, W149C did not allow transport to the substrates to be tested, while the other mutants retained significant transport ability. The reaction of the different mutant proteins with Fluorescein-NEM revealed that the mutant L146C was highly reactive with NEM; the W149C and S150C mutants were moderately reactive; A147C was barely reactive and V148C showed no reactivity. Conclusions The study identified that residues W149 and S150 situated at the interface of the aqueous： lipid junction as functionally important residues, probably involved in the substrate binding and translocation of QacA.