Jonas Michel Wolf

Cervical human papillomavirus infection and persistence: a clinic-based study in the countryside from South Brazil

Human papillomavirus (HPV) infection is common in sexually active women and viral persistence may cause intraepithelial lesions and eventually progress to cervical cancer (CC). The present study aimed to investigate epidemiological factors related to HPV infection and to evaluate viral persistence and CC precursor lesions frequencies in women from a city in the countryside of South Brazil. Three hundred women were recruited from a primary public health care clinic. The patients were interviewed and underwent sampling with cervical brushes for HPV-DNA detection/typing by a PCR-based assay and cytological analysis by Pap smear test. HPV was detected in 47 (15.7%) women. HPV infection was significantly associated with young age (<30 years) and low socio-economic status. Seventeen (5.7%) women presented cytological abnormalities, three of them with precursor CC intraepithelial lesions. A subgroup of 79 women had been previously analyzed and thirteen (16.4%) were persistently infected, two with precursor CC intraepithelial lesions and high-risk HPV types infection (both of them without cervical abnormalities in the first exam). In conclusion, HPV infection was associated with young age (<30 years) and low family income; viral persistence was low (16.4%) but related to CC precursor lesions; and HPV-DNA high risk types detection would help to screen CC in the population.

Performance of direct immunofluorescence assay for the detection of human metapneumovirus under clinical laboratory settings

INTRODUCTION Human metapneumovirus (hMPV) is an emergent human respiratory pathogen. This study aimed to evaluate the performance of direct immunofluorescence (DIF) to detect hMPV in a clinical laboratory setting. METHODS Nasopharyngeal aspirate samples (448) of children and adults with respiratory illness were used to detect hMPV by using DIF and real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays. RESULTS In all, 36 (8%) samples were positive by DIF and 94 (21%) were positive by qRT-PCR. Direct immunofluorescence specificity was 99% and sensitivity was 38%. CONCLUSIONS DIF is not very sensitive under clinical laboratory settings.

Human metapneumovirus in Southern Brazil

INTRODUCTION Infections caused by respiratory viruses are important problems worldwide, especially in children. Human metapneumovirus (hMPV) is a respiratory pathogen and causes severe infections with nonspecific symptoms. This study reports the hMPV occurrence and dissemination in southern Brazil and compares the frequency of occurrence of this virus and the human respiratory syncytial virus (hRSV) in the epidemiological weeks in a three-year period (2009-2011). METHODS In total, 545 nasopharyngeal (NP) specimens from individuals with Severe Acute Respiratory Syndrome (SARS) who were negative for other seven respiratory viruses were analyzed for the presence of hMPV. Human metapneumovirus was detected by direct immunofluorescence and real-time reverse transcription polymerase chain reaction. RESULTS hMPV was detected in 109 patients from the main geographic regions of the southernmost state of Brazil, presenting similar overall prevalence in males (46.8%) and females (53.2%). Among children who were less than six years old, hMPV was detected in 99 samples of all age groups, with a higher frequency in infants who were less than one year old (45.7%) compared to all other age groups until six years. hMPV and hRSV infection occurred in almost the same epidemiological weeks (EWs) of each year, with peaks of incidence between EW 31/37 and EW 26/38 for the years 2009 and 2011, respectively. hMPV was further detected in several cases of SARS and it was the only virus detected in three deaths. CONCLUSIONS These findings indicate that hMPV is in circulation in southern Brazil and highlight the importance of diagnosing hMPV for influenza-like illness in the population.

Polymorphisms in Genes Related to Cervical Cancer in A Brazilian Population: A Case-Control Study

To the editor: Cervical cancer (CC) is the fourth most common cancer in women, with approximately 528,000 new cases in the world each year, 80% of them in developing countries [1]. It is well recognized that persistent infection of human papillomavirus (HPV) is the main cause of precursor lesions that progress to CC, but only a small proportion of these HPV infected women develop the disease. In this sense, polymorphisms in human genes have also been associated with CC [2]. Genome-wide studies investigated the association of human single nucleotide polymorphisms (SNPs) with HPV persistent infection, progression to cervical intraepithelial neoplasia (CIN) and CC in Latin American women [3, 4]. More than seven thousand SNPs were investigated in genes related to immune response, DNA repair, viral replication and entry into the host cell. Association to persistent HPV progression to CIN and CC was observed with SNPs in genes of DNA repair (EXO1, CYBA, FANCA, XRCC1, GTF2H4, DUT, FLJ35220, and DMC1), immune response (IRF) and virus entry into the cell (SULF1 and OAS3) [3, 4]. The present case-control study evaluated the frequency of nine SNPs (all of them previously demonstrated to have significant association with HPV persistence and/or cancer) and the association with CC in a population in South Brazil. The selected SNPs were located in genes of DNA repair (rs4149963 in EXO1, rs3784621 in DUT, rs4603608 in FLJ35220 and rs2239359 in FANCA), immune response (rs7251 in the IRF), and virus entry into the host cell (rs4737999, rs10108002, rs4284050 in SULF1, and rs12302655 in OAS3). The population sample of this study was 109 CC patients (mean age 50.3 ± 14.3 years; range 25–88 years), recruited during treatment at the Center of High Complexity in Oncology (Centro de Assistência de Alta Complexidade em Oncologia CACON), located in the city of Ijuí in the Brazil’s southernmost state (Rio Grande do Sul), from 2012 to 2016; and 220 controls (mean 49.5 ± 13.2 years; range 21–82 years) recruited at the Women’s Health Center (Centro de Saúde da Mulher), a primary public health care clinic located in the city of Cruz Alta (also in Rio Grande do Sul State, Brazil) from 2012 to 2013. This last women group was previously characterized in cross-sectional epidemiological study [5]. Biological samples were obtained from the mouth in the CC patients (cases) and from the endocervix in the healthy controls. Buccal and endocervical cells were obtained by exfoliation using cytobrush and after stored in a buffer solution (EDTA pH = 8.0 0.01 M, SDS 0.03 M) at −20 °C until analysis. Total DNA was extracted from peripheral blood cells by silica adsorption method. EXO1 (rs4149963), DUT (rs3784621), FLJ35220 (rs4603608), FANCA (rs2239359), IRF3 (rs7251), SULF1 (rs4737999, rs10108002 and rs4284050) and OAS3 (rs12302655) SNPs were genotyped using TaqMan® specific SNP genotyping assays (Life Technologies Co, Carlsbab, CA, USA). Allelic discrimination real-time polymerase chain reactions (PCR) were performed on the StepOnePlusTM system according to conditions informed by this manufacturer. Thermal cycling conditions were: 10 min at 95 °C followed by 45 cycles of 15 s at 95 °C and 1 min at 60 °C. Allelic discrimination was performed by measuring end-point fluorescence using * Jonas Michel Wolf jonasmwolf@gmail.com

Discriminant analyzes for the use of tobacco, alcohol and other drugs among adolescents in an urban center in southern Brazil

Introduction : The consumption of tobacco, alcohol, and illicit drugs is a problem in adolescence, especially in the school context. This study aimed to identify the demographic and social variables that distinguished the students who had used tobacco, alcohol and/or other drugs at some point of their lives from those who had never used these substances. Methods : A school-based cross-sectional study was conducted to evaluate students attending the ninth year of elementary education in the city of Caxias do Sul (n = 1,285). Results : Alcohol was the most consumed drug (74.9%). In all the three discriminant profiles, adolescents who used tobacco throughout their lives presented more family conflict and hierarchy, higher age, greater difficulty in talking to their mother, father, and siblings, and higher school failure rates. The group that reported lifetime use of alcohol showed similar characteristics, and also reported greater loneliness. The group that has used illicit drugs was characterized, as well as other factors, by meeting with friends outside school more often, not having a good friend, being lonely, and having difficulties in talking to their father. Conclusion : Several social and family-related aspects can act as factors that propitiate adolescents from drugs. Public policies in this regard are of fundamental importance. Keywords : Adolescents; use of drugs; family; discriminant analysis

Increasing prevalence and dissemination of invasive nontyphoidal Salmonella serotype Typhimurium with multidrug resistance in hospitalized patients from southern Brazil

INTRODUCTION Nontyphoidal Salmonella serotypes are the main cause of human food-borne infection, including several hospitalization cases in the developing countries. AIM To detect the main serotypes and to characterize the antibiotic resistance of human non-enteric and enteric nontyphoidal Salmonella from clinical isolates in Brazil. METHODS Salmonella serotypes were identified by microbiological and molecular methods. Susceptibility testing to antibiotics was performed by agar disk diffusion. Real-time PCRs were carried out for the detection of the genus Salmonella as well as serotypes Typhimurium and Enteritidis. RESULTS A total of 307 nontyphoidal Salmonella were isolated from 289 different patients in a reference laboratory (LACEN-RS) from Southern Brazil in a six-year period (2010-2015). There were 45 isolates from emerging cases and 244 from sporadic cases in hospitalized patients. Non-enteric isolates were detected in 42.6% of the patients from sources such as urine, blood and other clinical fluids. Serological and PCR-specific tests demonstrated that Typhimurium (48.4%) and Enteritidis (18.3%) were the most frequent serotypes. Typhimurium isolates were generally resistant to three or more antibiotic classes, while Enteritidis isolates to one or two classes. Typhimurium was the most frequent serotype in all samples (48.4%), mainly among the hospitalized patients (55.6%), and presented the highest rates of multidrug resistance (59.3% of the isolates of this serotype). Further, the prevalence of this serotype increased along the years of the study in comparison to other nontyphoidal Salmonella serotypes. CONCLUSION Greater public health attention should be given to prevent salmonellosis in the community and in hospital settings to reduce the rates of Typhimurium strains with multidrug resistance.

HBV epidemiology and genetic diversity in an area of high prevalence of hepatitis B in southern Brazil

BACKGROUND Hepatitis B virus (HBV) infection is a major public health problem in Brazil. HBV endemicity is usually moderate to low according to geographic regions, and high prevalence of this virus has been reported in people of some specific Brazilian counties, including those with a strong influence of Italian colonization in southern Brazil. Analysis of HBV diversity and identification of the main risk factors to HBV infection are necessary to understand hepatitis B epidemiology in these high prevalence regions in southern Brazil. OBJECTIVE To investigate epidemiological characteristics and HBV genotypes and subgenotypes circulating in a specific city with high HBV prevalence. METHODS A cross-sectional study was performed with 102 HBV chronically infected individuals, recruited in reference outpatient clinics for viral hepatitis in a city of high HBV prevalence (Bento Gonçalves) in Rio Grande do Sul state, Brazil between July and December 2010. Socio-demographic, clinical and behavior-related variables were collected in a structured questionnaire. HBV serological markers (HBsAg, anti-HBc), viral load, genotypes/subgenotypes and drug resistance were evaluated and comparatively analyzed among all patients. RESULTS The HBV infected subjects had a mean age of 44.9 (±12.2) years, with 86 patients (84.3%) reporting to have a family history of HBV infection, 51 (50.0%) to share personal objects, and were predominantly of Italian descendants (61; 64.9%). There was a predominance of genotype D (49/54; 90.7%), but genotype A was also detected (5/54; 9.3%). Subgenotypes D1 (1; 4.7%), D2 (3; 14.3%), and D3 (17; 81.0%) were identified. LAM-resistant mutation (rtM204I) and ADV-resistant mutations (rtA181V) were detected in only one patient each. CONCLUSIONS These results demonstrate a pivotal role of intrafamilial transmission for HBV spreading in this population. Furthermore, there is a high prevalence of HBV genotype D in this region.

Risk factors for hepatitis B transmission in South Brazil

BACKGROUND Hepatitis B virus (HBV) infection is a major public health problem in Brazil. Several risk factors are involved in HBV infection and their identification by a rational and essential approach is required to prevent the transmission of this infection in Brazil. OBJECTIVES To evaluate risk factors associated with HBV infection in South Brazil. METHODS A total of 260 patients with HBV and 260 controls from Caxias do Sul (state of Rio Grande do Sul, Brazil) participated in this study. All participants were given a standard questionnaire to yield the sociodemographic information and to identify HBV risk factors. HBV infection was detected by HBsAg test in all participants. FINDINGS HBV infection in these cases was strongly associated with history of a family member HBV-infected, mainly mother [odds ratio (OR) = 4.86; 95% confidence intervals (CI): 1.69–13.91], father (OR = 5.28; 95% CI: 1.58–17.71), and/or siblings (OR = 22.16; 95% CI: 9.39–52.25); sharing personal objects (OR = 1.40; 95% CI: 1.37–2.38); and having history of blood transfusion (OR = 2.05; 95% CI: 1.10–2.84). CONCLUSIONS HBV infection was strongly associated with having a family member infected with hepatitis B, sharing personal objects, and having history of blood transfusion.

Lack of association between human leukocyte antigen polymorphisms rs9277535 and rs7453920 and chronic hepatitis B in a Brazilian population

Hepatitis B virus (HBV) infection is a serious public health problem worldwide. The progression of the disease depends on several host and viral factors and may result in fulminant hepatitis (very rare), acute hepatitis with spontaneous clearance, and chronic hepatitis B infection. Previous studies demonstrated that variations in the human leukocyte antigen (HLA) class II (HLA-DPB1 and HLA-DQB2 genes) are related to the chronic HBV infection. This study aimed to investigate the association of two single nucleotide polymorphism (SNPs), one in the HLA-DPB1 (rs9277535) and one in the HLA-DQB2 (rs7453920), with chronic hepatitis B infection in a southern Brazilian sample. This case-control study included 260 HBV patients attended in a Specialized 2 V.R.Z.B. Pereira et al. Genetics and Molecular Research 16 (2): gmr16029565 Center for Health in Caxias do Sul (Brazil) between 2014 and 2016. The same number of controls (matching for age, gender, and ethnicity) was obtained in a University Hospital in the same city and period. Blood samples were collected and genomic DNA was extracted. Genotyping were performed by real-time Taqman PCR method. Odds ratios with 95% confidence intervals and significance level of 5% (P < 0.05) were calculated. Allele frequencies in the SNP rs9277535 were 72.6% for A and 27.4% for G nucleotides in cases and 75.0% for A and 25.0% for G in controls. Allele frequencies in the SNP rs7453920 were of 25.7% for A and 74.3% for G in cases and 28.8% for A and 71.2% for G in controls. No statistically significant association was found between both SNPs and chronic hepatitis B (P > 0.05).

Fases pré-analítica, analítica e pós-analítica no monitoramento laboratorial da anticoagulação com antagonistas da vitamina K

A trombose e uma doenca caracterizada por eventos de hipercoagulabilidade. A terapeutica anticoagulante oral com antagonistas da vitamina K (AVKs) e amplamente indicada para prevencao e/ou controle de disturbios da coagulacao. O manuseio de administracao dos AVKs e dificil devido a complexidade da definicao da dose. Em geral, o monitoramento de individuos submetidos a terapeutica com AVK e realizado pela determinacao do tempo de protrombina, em que se avalia o grau de anticoagulacao atraves do coeficiente internacional normatizado. Invariavelmente, o fluxo do processamento laboratorial, que compreende as fases pre-analitica, analitica e pos-analitica, e importante para a fidedignidade dos resultados, repercutindo na conduta medica de forma determinante. O objetivo deste estudo foi a realizacao de uma revisao da literatura cientifica descritiva utilizando bases de dados eletronicos para busca de materiais cientificos, como Google Scholar, MEDLINE, LILACS, PubMed, SciELO e Science Direct. Abordamos alguns aspectos relacionados ao fluxo analitico da monitoracao laboratorial em um laboratorio de analises clinicas. Em conclusao, e necessaria uma apropriada conducao das fases analiticas para que seja possivel o sucesso terapeutico com uso dos AVKs. Palavras-chave: Tempo de protrombina; coeficiente internacional normatizado; fases analiticas