Mutations in the highly conserved tyrosine-methionine-aspartate-aspartate (YMDD) motif are frequently associated with resistance to antivirals and represent a major concern in the treatment of hepatitis B virus (HBV) infection. Conventional methods fail to detect minority populations of drug-resistant viral quasispecies if they represent less than 25% of the total sample virus population. The amplification refractory mutation system real-time PCR (ARMS RT-PCR) was combined with molecular beacon technology using the LightCycler system. The samples from HBV patients selected for assay evaluation included (i) 57 samples from treatment-naive patients for biological discriminatory ability (cutoff) estimation, (ii) 12 samples from patients with treatment failure that were M204V positive by sequencing, and (iii) 13 samples from patients with treatment failure that were negative for mutation at codon 204 by sequencing. The discriminatory ability of the assay was 0.25% when tested with laboratory-synthesized DNA target sequences. The median mutant-to-wild-type ratio for samples from naive patients tested positive for the wild type and for mutant variants was 0.01% (5th and 95th percentiles = 0.0001 and 0.04%, respectively). A value of 0.04% was selected as the biological cutoff of the assay of clinical samples. In all samples M204V positive by sequencing (12/12), the mutant variant was detected as the predominant population (range, 82.76 to 99.43%). Interestingly, in 5 (38%) of 13 samples negative by sequencing, the M204V variant was detected at a ratio above the biological cutoff (0.05 to 28%). The assay represents an efficient technique for the early detection and quantification of M204V variants before mutant strains emerge to dominate the population.

BACKGROUND: The effect of hepatitis B virus (HBV) infection on the natural history of human immunodeficiency virus (HIV) disease remains uncertain. Therefore, a retrospective cohort study was conducted to examine the influence of HIV-HBV coinfection on AIDS development and overall mortality. Moreover, our results were added to those of previous studies in a literature-based meta-analysis. METHODS: Serum samples obtained from HIV-seropositive patients from 1984 through 2003 were retrospectively tested for hepatitis B surface antigen. Multivariable analyses were performed using Poisson and logistic regression models. For meta-analytic purposes, eligible articles were identified and relevant data were abstracted. Pooled estimates of effect were calculated applying fixed and random effects models. RESULTS: The prevalence of chronic HBV infection (documented hepatitis B surface antigen seropositivity for >6 months) among 1729 HIV-positive patients was approximately 6%. The multivariable analyses in our primary study revealed no significant impact of concomitant HIV-HBV infection on progression to AIDS and all-cause mortality. However, a meta-analysis performed on data from 12,382 patients enrolled in 11 studies revealed a significant effect of HIV-HBV coinfection on overall mortality (pooled effect estimate, 1.36; 95% confidence interval, 1.12-1.64). The increased rate of death among coinfected individuals was observed in the meta-analyses of studies conducted both before (pooled effect estimate, 1.60; 95% confidence interval, 1.07-2.39) and after (pooled effect estimate, 1.28; 95% confidence interval, 1.03-1.60) commencement of highly active antiretroviral therapy. CONCLUSIONS: HIV-HBV coinfection seems to affect all-cause mortality, and strategies to reduce liver damage in patients coinfected with HIV and HBV are justified.

OBJECTIVE: Skin cancer campaigns have only recently been implemented in a wide scale in Greece. In order to better target those individuals at risk, we aimed to explore sun exposure behaviors in a sample of the Greek population. METHODS: A total of 802 subjects, randomly selected among the residents of the two largest Greek cities, underwent a questionnaire-based telephone interview, assessing the degree of sun exposure patterns and knowledge of skin cancer risk factors. RESULTS: Females more commonly (90%) used sunscreens compared with males (67.5%), and among sunscreen users, only 8% used products with SPF <10. Almost half of the participants (48%) preferred going to the beach during the midday hours. Over exposure to the sun was mostly reported by individuals with phototypes III/IV and of younger age (18-35 years). Most participants were aware of sun exposure as a cause of skin cancer, but among those who did not, 41% were younger than 34 years of age. CONCLUSIONS: In our survey, significant differences in age, sex and phototypes were observed with regard to sun exposure patterns and sunscreen use. Educational campaigns should focus more on younger age groups, encouraging a more complete set of photoprotective measures. However, potential trial limitations inherent in self-reported measures should be taken into account.

A simple, safe and cost-effective treatment protocol in ovarian stimulation is of great importance in IVF practice, especially in the case of previous unsuccessful attempts. hCG has been used as a substitute of LH because of the degree of homology between the two hormones. The main aim of this prospective randomized study was to determine, for the first time, whether low dose hCG added to rFSH for ovarian stimulation could produce better results compared to the addition of rLH in women entering IVF-ET, especially in those women that had previous IVF failures. An additional aim was to find an indicator that would allow us to follow-up ovarian stimulation and, possibly, modify it in order to achieve a better IVF outcome; and that indicator may be the cDNA copies of the LH/hCG receptor. Group A patients (n = 58) were administered hCG and Group B rLH (n = 56) in addition to rFSH in the first days of ovarian stimulation. The number of follicles and oocytes and, most importantly, implantation and pregnancy rates were shown to be statistically significantly higher in the hCG group. This study has also determined, for the first time to our best knowledge, m-RNA for LH/hCG receptors in the lymphocytes of peripheral blood 40 h before ovum pick-up. cDNA levels of the hCG receptor after ovarian stimulation were significantly higher among women receiving hCG compared to those receiving LH. In addition, higher levels were encountered among women with pregnancy compared to those without, although this was not statistically significant due to the small number of pregnancies. It seems that hCG permits a highly effective and more stable occupancy of rLH/hCG receptors and gives more follicles and more oocytes. The determination of cDNA copies could be, in the future, a marker during ovulation induction protocols and of course a predictor for the outcome of ART in the special subgroup of patients with previous failures.

The SLC24A5 gene, the human orthologue of the zebrafish golden gene, has been shown to play a key role in human pigmentation. In this study, we investigate the prevalence of the variant allele rs1426654 in a selected sample of Greek subjects. Allele-specific polymerase chain reaction was performed in peripheral blood samples from 158 attendants of a dermatology outpatient service. The results were correlated with pigmentary traits and MC1R genotype. The vast majority of subjects (99%) were homozygous for the Thr(111) allele. Only two subjects from the control group (1.26%) were heterozygous for the alanine and threonine allele. Both of these Thr(111)/Ala(111) heterozygotes carried a single polymorphism of MC1R (one with the V92M variant and another with the V60L variant). Following reports of the rs1426654 polymorphism reaching fixation in the European population, our study of Greek subjects showed a prevalence of the Thr(111) allele, even among subjects with darker skin pigmentation or phototype.

Previous studies showed that high levels of human immunodeficiency virus type 1 (HIV-1) DNA are associated with a faster progression to AIDS, an increased risk of death, and a higher risk of HIV RNA rebound in patients on highly active antiretroviral therapy. Our objective was to develop and assess a highly sensitive real-time multiplex PCR assay for the quantification of HIV-1 DNA (RTMP-HIV) based on molecular beacons. HIV-1 DNA quantification was carried out by RTMP in a LightCycler 2.0 apparatus. HIV-1 DNA was quantified in parallel with CCR5 as a reference gene, and reported values are numbers of HIV-1 DNA copies/10(6) peripheral blood mononuclear cells (PBMCs). The clinical sensitivity of the assay was assessed for 115 newly diagnosed HIV-1-infected individuals. The analytical sensitivity was estimated to be 12.5 copies of HIV-1 DNA per 10(6) PBMCs, while the clinical sensitivity was 100%, with levels ranging from 1.23 to 4.25 log(10) HIV-1 DNA copies/10(6) PBMCs. In conclusion, we developed and assessed a new RTMP-HIV assay based on molecular beacons, using a LightCycler 2.0 instrument. This multiplex assay has comparable sensitivity, reproducibility, and accuracy to single real-time PCR assays.

The impact of prioritization and of timing of vaccination strategies on reducing transmission of pandemic influenza A(H1N1) was evaluated in a community with the structure of the Greek population using a stochastic simulation model. Prioritization scenarios were based on the recommendations of the United States Centers for Disease Control and Prevention Advisory Committee on Immunization Practices and vaccination was assumed to initiate either before or during the ongoing epidemic. In the absence of intervention, an illness attack rate (AR) of 34.5% is anticipated. Vaccinating the priority groups before the epidemic (pregnant women, people who live with or care for children <6 months of age, healthcare/emergency services personnel, children 6 months-4 years old and high-risk children 5-18 years old) will have a negligible impact on the overall AR. Vaccinating the recommended groups before the epidemic (priority groups as well as all persons 6 months-24 years old and high-risk individuals 25-64 years old) is anticipated to result in overall and age-specific ARs within the range of seasonal influenza (5%-15%). Initiating vaccination early during the epidemic (AR

A cross-sectional telephone survey on a nationally representative sample of 1,000 Greek households was performed to assess the acceptability of the pandemic influenza A(H1N1)v vaccine, factors associated with intention to decline and stated reasons for declining vaccination. The survey was initiated the last week of August 2009 (week 35) and is still ongoing (analysis up to week 44). The percentage of participants answering they would probably not/definitely not accept the vaccine increased from 47.1% in week 35 to 63.1% in week 44 (test for trend: p<0.001). More than half of the people which chronic illnesses (53.3%) indicated probably not/definitely not. Factors associated with intention to decline vaccination were female sex, age between 30-64 years, perception of low likelihood of getting infected or of low risk associated with influenza, and absence of household members suffering from chronic illnesses. For the majority of the respondents (59.8%), the main reason for intending to decline vaccination was the belief that the vaccine might not be safe. Promotion of vaccination programmes should be designed taking into account the attitudinal barriers to the pandemic vaccine.

BACKGROUND: The evaluation of academic research performance is nowadays a priority issue. Bibliometric indicators such as the number of publications, total citation counts and h-index are an indispensable tool in this task but their inherent association with the size of the research output may result in rewarding high production when evaluating institutions of disparate sizes. The aim of this study is to propose an indicator that may facilitate the comparison of institutions of disparate sizes. METHODS: The Modified Impact Index (MII) was defined as the ratio of the observed h-index (h) of an institution over the h-index anticipated for that institution on average, given the number of publications (N) it produces i.e. MII = h/10alphaNbeta (alpha and beta denote the intercept and the slope, respectively, of the line describing the dependence of the h-index on the number of publications in log10 scale). MII values higher than 1 indicate that an institution performs better than the average, in terms of its h-index. Data on scientific papers published during 2002-2006 and within 36 medical fields for 219 Academic Medical Institutions from 16 European countries were used to estimate alpha and beta and to calculate the MII of their total and field-specific production. RESULTS: From our biomedical research data, the slope beta governing the dependence of h-index on the number of publications in biomedical research was found to be similar to that estimated in other disciplines ( approximately 0.4). The MII was positively associated with the average number of citations/publication (r = 0.653, p < 0.001), the h-index (r = 0.213, p = 0.002), the number of publications with > or = 100 citations (r = 0.211, p = 0.004) but not with the number of publications (r = -0.020, p = 0.765). It was the most highly associated indicator with the share of country-specific government budget appropriations or outlays for research and development as % of GDP in 2004 (r = 0.229) followed by the average number of citations/publication (r = 0.153) whereas the corresponding correlation coefficient for the h-index was close to 0 (r = 0.029). MII was calculated for first 10 top-ranked European universities in life sciences and biomedicine, as provided by Times Higher Education ranking system, and their total and field-specific performance was compared. CONCLUSION: The MII should complement the use of h-index when comparing the research output of institutions of disparate sizes. It has a conceptual interpretation and, with the data provided here, can be computed for the total research output as well as for field-specific publication sets of institutions in biomedicine.

Mathematical modeling was employed on recent epidemiological data from Mexico in order to assess the impact of intervention strategies on the spread of influenza A(H1N1)v in the setting of the European region. Active surveillance that will ensure timely treatment and home isolation of symptomatic cases in combination with school closure seem to form an efficient strategy to control the spread of influenza A(H1N1)v.

The aim of the study was to evaluate the prevalence and risk factors of HPV in a gynecologic population attending outpatient clinics using two new molecular tests. The Amplicor HPV test and the Linear Array (LA) HPV Genotyping test were used for the detection of HPV DNA in 320 women. Multiple logistic regression was used to identify independent prognostic factors of HPV positivity. The agreement between the two methods in terms of their qualitative results was 89.3% (kappa: 0.63). Based on the LA results, the overall prevalence of HPV DNA was 49.1%, 95% confidence interval (95% CI: 43.5%, 54.7%). The prevalence of high-risk HPV types was 30.3%. The predominant types were HPV-6 (24.8%) and HPV-16 (20.4%). Among women with normal cytology, the prevalence of HPV was much higher in those presenting other findings, such as inflammation, than those without other abnormal findings (49.5% vs. 31.5%). On the basis of multivariate analysis, the risk of HPV infection was higher among women with multiple sexual partners [>3 vs. 1: OR = 3.1, 95% CI: (1.5, 7.2)], Pap smear findings [low/high-grade lesions vs. negative: OR = 2.8, 95% CI: (1.2, 6.5)], the presence of warts [yes vs. no: OR = 3.0, 95% CI: (1.5, 6.3)] and no history of child birth [no vs. yes: OR = 2.6, 95% CI: (1.0, 6.7)]. Younger age was an additional risk factor for HPV infection with carcinogenic genotypes [OR for 1 year increase = 0.93, 95% CI: (0.89, 0.98)].