Molecular characterization of Chikungunya virus recovered from patients in the Maranhão state, Brazil.

BACKGROUND: Chikungunya virus (CHIKV) is an arbovirus from the Togaviridae family which has four genotypes: West African (WA), East/Central/South African (ECSA) and Asian/Caribbean lineage (AL) and Indian Ocean Lineage (IOL). The ECSA genotype was first registered in Brazil in Feira de Santana and spread to all Brazilian regions. This study reports the characterization of CHIKV isolates recovered from sera samples of fifty patients from seventeen cities in Maranhão, a state from Brazilian northeast region and part of the Legal Amazon area. METHODS AND RESULTS: Primers were developed to amplify the partial regions coding structural proteins (E1, E3, E2, 6 K, and Capsid C). The consensus sequences have 2871 bp, covering approximately 24% of the genome. The isolates were highly similar (> 99%) to the ECSA isolate from Feira de Santana (BHI3734/H804698), presenting 30 non-synonymous mutations in E1 (5.95%), 18 in E2 (4.46%), and 1 in E3 (3.03%), taking the BHI3734/H804698 isolate as standard. Although the mutations described have not previously been related to increased infectivity or transmissibility of CHIKV, in silico analysis showed changes in physicochemical characteristics, antigenicity, and B cell epitopes of E1 and E2. CONCLUSIONS: These findings demonstrate the importance of molecular approaches for monitoring the viral adaptations undergone by CHIKV and its geographic distribution.

Deaths by COVID-19 in a hospital from a city from Legal Amazon region: what lessons could be learned? / Mortes por COVID-19 em um hospital de uma cidade da região da Amazônia Legal: que lições podem ser aprendidas?

Objective: Describe the epidemiology of COVID-19 deaths within a hospital in the Amazon region in a period of 64 days, which corresponds to the growth curve of the COVID-19 first-wave pandemic in 2020. Methods: The data were obtained from medical records of 152 deaths registered for adults and elderly hospitalized. The data were also compared with the number of deaths in previous years during the same period studied to assess the impact of the pandemic on this hospital. The study also assesses the impact of intra-hospital transfers, accounting for the number of times patients who died performed transfers between sectors of the hospital. Results: During the period analyzed, there was an increase in deaths compared to the previous years. The majority of dead patients were male, aged between 34 and 96 years. The deaths were associated comorbidities such as arterial hypertension, diabetes mellitus, and kidney disease. The SARS-CoV-2 infection was confirmed in 91 cases. Among them, 15 individuals were admitted without conditions related to SARS-CoV-2 infection; they had a three-fold higher number of hospital transfers than those admitted with SARS-CoV-2 infection symptoms. Sixteen patients with SARS-CoV-2 infection developed respiratory symptoms just after hospitalization. The diagnostic exam for SARS-CoV-2 infection was performed on average 4 (± 6) days after the onset of symptoms and 6 (± 6) days after admission, and the average time from the onset of respiratory symptoms to death was 4 (± 6) days. Conclusions: These data suggest the high presence of hospital infection by SARS-CoV-2 in the Brazilian Amazon region, which may be related to the number of sectorial transfers, delay in confirming the diagnosis, and lack of management. We report a serious public health problem, as it demonstrates the fragility of healthcare institutions in the hospital environmen (AU).

Objetivo: Descrever a epidemiologia de mortes por COVID-19 em um hospital na região da Amazônia em um período de 64 dias, que corresponde à curva de crescimento da primeira onda da pandemia de COVID-19 em 2020. Métodos: Os dados foram obtidos de 152 óbitos registrados em prontuários de adultos e idosos hospitalizados. Os dados foram também comparados com o número de óbitos em anos anteriores, no mesmo período estudado, de forma a avaliar o impacto da pandemia neste hospital. O estudo também avalia o impacto das transferências intra-hospitalares, contabilizando o número de vezes que os pacientes que faleceram realizaram transferências entre setores do hospital. Resultados: No período analisado, houve aumento de óbitos em relação aos anos anteriores. A maioria dos pacientes mortos era do sexo masculino, com idade entre 34 e 96 anos. Os óbitos foram associados a comorbidades como hipertensão arterial, diabetes mellitus e doença renal. A infecção por SARS-CoV-2 foi confirmada em 91 casos. Entre eles, 15 indivíduos foram internados sem condições relacionadas à infecção por SARS-CoV-2; eles tiveram um número três vezes maior de transferências hospitalares do que aqueles admitidos com sintomas de infecção por SARS-CoV-2. Dezesseis pacientes com infecção por SARS-CoV-2 desenvolveram sintomas respiratórios logo após a hospitalização. O exame diagnóstico para infecção por SARS-CoV-2 foi realizado em média 4 (± 6) dias após o início dos sintomas e 6 (± 6) dias após a admissão, e o tempo médio do início dos sintomas respiratórios até o óbito foi de 4 ( ± 6) dias. Conclusões: Esses dados sugerem alta presença de infecção hospitalar por SARS-CoV-2 na região amazônica brasileira, o que pode estar relacionado ao número de transferências setoriais, demora na confirmação do diagnóstico e falta de manejo. Relatamos um grave problema de saúde pública, pois demonstra a fragilidade das instituições de saúde no ambiente hospitalar (AU).

Human protothecosis: A case report in northeastern Brazil.

We describe a case of extensive human cutaneous protothecosis in the northeastern of Brazil. The diagnosis was made through histopathological examination of skin samples and culturing on Sabouraud dextrose agar, with identification through mass spectrometry. Treatment with itraconazole failed and was switched to liposomal amphotericin followed by voriconazole with clinical cure. Prothotecosis should be a diagnostic hypothesis when there are skin lesions after trauma, specially in situations of atypical evolution.

Characterization, antimicrobial and cytotoxic activity of polymer blends based on chitosan and fish collagen.

This study aims to produce, characterize, and assess the antimicrobial activity and cytotoxicity of polymer blends based on chitosan (CT) and fish collagen (COL) produced by different precipitation methods. Polymer blends were obtained in alkaline (NaOH), saline (NaCl), and alkaline/saline (NaOH/NaCl) solutions with different CT:COL concentration ratios (20:80, 50:50, and 80:20). The polymer blends were characterized by various physicochemical methods and subsequently evaluated in terms of their in vitro antimicrobial and cytotoxicity activity. In this study, the degree of chitosan deacetylation was 82%. The total hydroxyproline and collagen content in the fish matrix was 47.56 mg. g-1 and 394.75 mg. g-1, respectively. The highest yield was 44% and was obtained for a CT:COL (80:20) blend prepared by precipitation in NaOH. High concentrations of hydroxyproline and collagen in the blends were observed when NaOH precipitation was used. Microbiological analysis revealed that the strains used in this work were sensitive to the biomaterial; this sensitivity was dose-dependent and increased with increasing chitosan concentration in the products. The biocompatibility test showed that the blends did not reduce the viability of fibroblast cells after 48 h of culture. An analysis of the microbiological activity of the all-polymer blends showed a decrease in the values of minimal inhibitory concentration (MIC) and minimal bactericidal concentrations (MBC) for S. aureus and P. aeruginosa. The blends showed biocompatibility with NIH-3T3 murine fibroblast cells and demonstrated their potential for use in biomedical applications such as wound healing, implants, and scaffolds.

Molecular Detection of Drug-Resistance Genes of blaOXA-23-blaOXA-51 and mcr-1 in Clinical Isolates of Pseudomonas aeruginosa.

Pseudomonas aeruginosa has caused high rates of mortality due to the appearance of strains with multidrug resistance (MDR) profiles. This study aimed to characterize the molecular profile of virulence and resistance genes in 99 isolates of P. aeruginosa recovered from different clinical specimens. The isolates were identified by the automated method Vitek2, and the antibiotic susceptibility profile was determined using different classes of antimicrobials. The genomic DNA was extracted and amplified by multiplex polymerase chain reaction (mPCR) to detect different virulence and antimicrobial resistance genes. Molecular typing was performed using the enterobacterial repetitive intergenic consensus (ERIC-PCR) technique to determine the clonal relationship among P. aeruginosa isolates. The drug susceptibility profiles of P. aeruginosa for all strains showed high levels of drug resistance, particularly, 27 (27.3%) isolates that exhibited extensively drug-resistant (XDR) profiles, and the other isolates showed MDR profiles. We detected the polymyxin E (mcr-1) gene in one strain that showed resistance against colistin. The genes that confer resistance to oxacillin (blaOXA-23 and blaOXA-51) were present in three isolates. One of these isolates carried both genes. As far as we know from the literature, this is the first report of the presence of blaOXA-23 and blaOXA-51 genes in P. aeruginosa.

The effect of family history of hypertension and polymorphism of the ACE gene (rs1799752) on cardiac autonomic modulation in adolescents.

This study aims to analyze the behaviour of cardiac autonomic modulation in adolescents with a family history of hypertension along with polymorphism of the ACE gene (rs1799752). The study involved 141 adolescents, with a mean age of 14.89, divided into the following six groups: offspring of normotensive parents (ONP): DD, DI and II; and offspring of hypertensive parents (OHP): DD, DI and II. Blood pressure, body composition, family history of hypertension, sleep disorder, and sexual maturation were assessed for the groups' characterization. Afterwards, an electrocardiogram was performed, and oral mucosal cells were collected to analyze heart rate variability and genotypic research of angiotensin-converting enzyme. The main finding of this study was the decrease of vagal action in group OHP (genotype DD) relative to group ONP (genotype II): LF (%), 54.25 ± 3.14 vs 39.33 ± 3.80; HF (%), 45.74 ± 3.14 vs 60.66 ± 3.80; LF/HF, 1.48 ± 0.23 vs 0.68 ± 0.19. The results also showed changes in the variable diastolic blood pressure (DBP) in OHP (genotype DI) to ONP (genotype II) groups: 72.99 ± 2.33 vs 63.27 ± 1.72; and OHP (genotype DI) to ONP (genotype DD) groups. Adolescents with genotype DD and a family history of arterial hypertension present chances in cardiac autonomic modulation, the cardiac parasympathetic modulation is lower in these adolescents in comparison to participants of ONP + II group.

Salivary biomarkers for caries susceptibility and mental stress in individuals with facial pain.

Objective: To assess salivary biomarkers for dental caries susceptibility and mental stress in young adults with perceived facial pain.Methods: Males and females who reported facial pain and pain-free controls participated in this study. Facial pain was investigated using the RDC/TMD. Unstimulated saliva was then collected for the evaluation of salivary flow rate (SFR), pH, Streptococcus mutans counts, morning cortisol, and S-IgA.Results: Women with facial pain had significantly lower SFR values, and the facial pain group showed different correlations among biomarkers for caries susceptibility and cortisol levels when compared to controls. Notably, higher SFR values were associated with a lower likelihood of having facial pain.Conclusion: Differences in SFR values, particularly in women, and markedly distinct interactions among the salivary biomarkers analyzed were observed between individuals with facial pain and pain-free controls. Hence, a connection between the dynamics of saliva, stress response, and facial pain perception might exist.

Identification and seasonality of rhinovirus and respiratory syncytial virus in asthmatic children in tropical climate.

INTRODUCTION: Asthma is a disease that has been associated with the presence of different genetic and socio-environmental factors. OBJECTIVE: To identify and evaluate the seasonality of respiratory syncytial virus (RSV) and human rhinovirus (RV) in asthmatic children and adolescents in tropical climate, as well as to assess the socioeconomic and environmental factors involved. METHODS: The study was conducted in a referral hospital, where a total of 151 children were recruited with a respiratory infection. The International Study of Asthma and Allergies in Childhood (ISAAC) protocol and a questionnaire were applied, and a skin prick test was performed. The nasal swab was collected to detect RV and RSV through molecular assay. National Meteorological Institute (INMET) database was the source of climatic information. RESULTS: The socio-environmental characterization of asthmatic children showed the family history of allergy, disturbed sleep at night, dry cough, allergic rhinitis, individuals sensitized to at least one mite. We identified RV in 75% of children with asthma and 66.7% of RSV in children with asthma. There was an association between the presence of RV and the dry season whereas the presence of the RSV was associated with the rainy season. Contributing to these results, a negative correlation was observed between the RSV and the wind speed and the maximum temperature (T. Max) and a positive correlation with precipitation. CONCLUSIONS: The results suggest a high prevalence of RV and RSV in asthmatic children and the seasonality of these viruses were present in different climatic periods. This has significant implications for understanding short- and long-term clinical complications in asthmatic patients.

Molecular detection of human T cell lymphotropic virus type 1 in pregnant women from Maranhão state, Brazil.

The human T cell lymphotropic virus (HTLV) has a worldwide distribution. HTLV is endemic in some states in the northeastern region of Brazil. This study investigated the prevalence of HTLV-1/2 in 713 pregnant women attended at the Central Laboratory of Public Health of Maranhão (LACEN-MA) between February 2015 and May 2017. Serological screening was performed by chemiluminescent microparticle immunoassay (CMIA), and reactive sera were subsequently confirmed by Western blot (WB) analysis. Five samples were determined to be HTLV-1/2-reactive by CMIA analysis, while in the WB analysis, three sera were positive for HTLV-1, and two were indeterminate. The polymerase chain reaction (PCR) analysis used to detect HTLV-1 proviral DNA showed a specific 336 base pair fragment for HTLV-1 in all CMIA-reactive serum samples. PCR products were purified and sequenced. We observed a 0.7% molecular prevalence of HTLV-1 infection. The average age of the HTLV-1-positive pregnant women was 25.6 ± 8.2 years, and the average age of the HTLV-1-negative pregnant women was 24.3 ± 6.2 (p = 0.60). We observed that there was no association of HTLV-1 infection with age, ethnicity, marital status, educational level, family income, age of first sexual intercourse, previous pregnancy, breastfeeding, intravenous drug use by partner, history of blood transfusions, or use of condoms. The prevalence of HTLV-1 observed in pregnant women demonstrated the need to implement public health policies for the screening of HTLV-1/2 in prenatal care and counseling to avoid breastfeeding by infected women; this approach could control vertical transmission and reduce the spread of this virus in the population.

Evaluation of the interaction between polymyxin B and Pseudomonas aeruginosa biofilm and planktonic cells: reactive oxygen species induction and zeta potential.

BACKGROUND: Although the most widely accepted mechanism of action for polymyxins is related to bacterial lysis via disruption, we hypothesized that this antimicrobial drug class could have other effects on Pseudomonas aeruginosa planktonic and sessile cells. Little is known regarding oxidative burst and zeta potential (ZP) data associated with the interaction between polymyxin B and P. aeruginosa cells. The present study evaluated endogenous reactive oxygen species (ROS) production and changes in the net charges of biofilm and planktonic cells in response to polymyxin B. RESULTS: Polymyxin B induced concentration-dependent killing at all concentrations tested in planktonic and sessile cells from P. aeruginosa strains. Sublethal concentrations of polymyxin B induced oxidative burst. ROS production was higher in resistant planktonic cells than in biofilm cells but this was not observed for susceptible cells. Moreover, no net surface charge alterations were observed in planktonic cells from a susceptible strain treated with polymyxin B, but a significant increase of ZP was noted in planktonic cells from a resistant strain. CONCLUSION: Oxidative burst generated by planktonic and sessile cells from P. aeruginosa strains against polymyxin B indicates that ROS may have an important role in the mechanism of action of this drug. ZP data revealed that electrostatic interactions of the cationic peptide with the anionic surface of the cells are strain-dependent. Therefore, we suggested that the intracellular effects of polymyxin B should be further investigated to understand polymyxin B-induced stress in P. aeruginosa.

Dengue e Chikungunya: soroepidemiologia em usuários da atenção básica / Dengue and chikungunya: seroepidemiology in primary care users

Objetivo: caracterizar o perfil epidemiológico das arboviroses dengue e Chikungunya. Método: trata-se de um estudo epidemiológico, descritivo, retrospectivo, quantitativo. Escolheu-se um total de mil prontuários de pacientes com sorologia IgM positiva para dengue e Chikungunya de forma aleatória entre os resultados sorológicos ocorridos no ano de 2016. Coletaram-se as informações sociodemográficas dos pacientes, e os resultados se apresentam em forma de figuras. Resultados: revela-se que, dos mil prontuários escolhidos aleatoriamente de ambas as arboviroses, o sexo feminino foi o mais acometido; a faixa etária com maior número de casos foi de 21 a 40 anos e a com menor número foi em maiores de 60 anos; a etnia parda e a zona urbana foram as que mais apresentaram casos de dengue e Chikungunya e, em relação à notificação, somente um pouco mais da metade dos casos foi notificado. Conclusão: apontou-se, pelo estudo, um perfil endêmico de infecção por arboviroses que merece atenção e esforço dos gestores, profissionais e da sociedade, para o devido controle e monitoramento das arboviroses no Brasil. Ressalta-se a necessidade de os serviços de Atenção Primária à Saúde terem resolutividade quanto à vigilância epidemiológica, principalmente na notificação e investigação das arboviroses de forma adequada.(AU)

Objective: to characterize the epidemiological profile of arboviruses dengue and Chikungunya. Method: this is an epidemiological, descriptive, retrospective, quantitative study. A total of one thousand patient records were selected from patients with dengue-positive IgM seropositivity and were randomly selected from the serological results of the year 2016. Sociodemographic information was collected from the patients, and the results are presented in the form of figures. Results: it is revealed that, of the thousand records chosen at random from both arboviruses, the female was the most affected; the age group with the highest number of cases was between 21 and 40 years old, and the smallest age group was older than 60 years; the ethnic group and the urban area were the ones that presented the most cases of dengue and Chikungunya, and in relation to the notification, only a little more than half of the cases were reported. Conclusion: an endemic profile of arbovirus infection that deserves attention and effort by managers, professionals and society, for the proper control and monitoring of arboviruses in Brazil, was pointed out by the study. It is important to emphasize the need for Primary Health Care services to have a high level of epidemiological surveillance, especially when reporting and investigating arboviruses in an adequate manner.(AU)

Objetivo: caracterizar el perfil epidemiológico de las arbovirosis dengue y chikungunya. Método: se trata de un estudio epidemiológico, descriptivo, retrospectivo, cuantitativo. Se eligió un total de mil prontuarios de pacientes con serología IgM positiva para dengue y chikungunya de forma aleatoria entre los resultados serológicos ocurridos en el año 2016. Se recogen las informaciones sociodemográficas de los pacientes y los resultados se presentan en forma de figuras. Resultados: se revela que, de los mil prontuarios elegidos aleatoriamente de ambas arbovirosis, el sexo femenino fue el más acometido; el grupo de edad con mayor número de casos fue de 21 a 40 años y la con menor número fue en mayores de 60 años; la etnia parda y la zona urbana fueron las que más presentaron casos de dengue y chikungunya y, en relación a la notificación, sólo un poco más de la mitad de los casos fue notificado. Conclusión: se apuntó, por el estudio, un perfil endémico de infección por arbovirosis que merece atención y esfuerzo de los gestores, profesionales y de la sociedad, para el debido control y monitoreo de las arbovirosis en Brasil. Se resalta la necesidad de que los servicios de Atención Primaria a la Salud tengan resolución en cuanto a la vigilancia epidemiológica, principalmente en la notificación e investigación de las arbovirosis de forma adecuada.(AU)

Sex Differences in Salivary Parameters of Caries Susceptibility in Healthy Individuals.

PURPOSE: To investigate salivary parameters of caries susceptibility in men and women in order to identify potential variations due to sexual dimorphism. MATERIALS AND METHODS: A total of 46 female and 24 male patients, aged 18-40 years, participated in this study. Unstimulated saliva was collected for the evaluation of flow rate, pH, secretory IgA, Snyder test scores, and Streptococcus mutans counts (confirmed by PCR assay). Statistical analysis included the Mann-Whitney, Kruskal-Wallis and Bonferroni tests, one-way ANOVA, and the Spearman correlation at a 5% significance level, followed by a general linear model and multiple linear regressions. RESULTS: Female participants presented lower salivary pH values compared to males (p < 0.05), and different patterns of correlation among salivary parameters were found in men (p < 0.05) and women (p < 0.001). When comparing the variables according to Snyder test scores in men and women, there was a significant difference for S. mutans levels in the male group, and for pH and IgA in the female group (p < 0.05). Gender was found to be a predictor of salivary flow (R2 = 0.05; p < 0.05) and pH (R2 = 0.16; p < 0.001). In the female group, multiple regression showed several predictors for salivary flow rate, pH, IgA and Snyder test scores (p < 0.05), whereas no predictor was found in the male group (p > 0.05). CONCLUSION: Different salivary patterns were observed in men and women, thus the implications of such findings for caries susceptibility require further investigation.

Himatanthus drasticus Leaves: Chemical Characterization and Evaluation of Their Antimicrobial, Antibiofilm, Antiproliferative Activities.

Plant-derived products have played a fundamental role in the development of new therapeutic agents. This study aimed to analyze antimicrobial, antibiofilm, cytotoxicity and antiproliferative potentials of the extract and fractions from leaves of Himatanthusdrasticus, a plant from the Apocynaceae family. After harvesting, H. drasticus leaves were macerated and a hydroalcoholic extract (HDHE) and fractions were prepared. Antimicrobial tests, such as agar-diffusion, Minimum Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were carried out against several bacterial species. Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogenes and Klebsiella pneumoniae were inhibited by at least one extract or fraction in the agar-diffusion assay (inhibition halos from 12 mm to 30 mm). However, the lowest MIC value was found for HDHE against K. pneumoniae. In addition, HDHE and its fractions were able to inhibit biofilm formation at sub-inhibitory concentrations (780 µg/mL and 1.56 µg/mL). As the best activities were found for HDHE, we selected it for further assays. HDHE was able to increase ciprofloxacin (CIP) activity against K. pneumoniae, displaying synergistic (initial concentration CIP + HDHE: 2 µg/mL + 600 µg/mL and 2.5 µg/mL + 500 µg/mL) and additive effects (CIP + HDHE: 3 µg/mL + 400 µg/mL). This action seems to be associated with the alteration in bacterial membrane permeability induced by HDHE (as seen by propidium iodide labeling). This extract was non-toxic for red blood cell or human peripheral blood mononuclear cells (PBMCs). Additionally, it inhibited the lipopolysaccharide-induced proliferation of PBMCs. The following compounds were detected in HDHE using HPLC-ESI-MS analysis: plumieride, plumericin or isoplumericin, rutin, quercetin and derivatives, and chlorogenic acid. Based on these results we suggest that compounds from H. drasticus have antimicrobial and antibiofilm activities against K. pneumoniae and display low cytotoxicity and anti-proliferative action in PBMC stimulated with lipopolysaccharide.

Phenotypic and molecular detection of the bla KPC gene in clinical isolates from inpatients at hospitals in São Luis, MA, Brazil.

BACKGROUND: Bacteria that produce Klebsiella pneumoniae carbapenemases (KPCs) are resistant to broad-spectrum ß-lactam antibiotics. The objective of this study was to phenotypically and genotypically characterize the antibiotic susceptibility to carbapenems of 297 isolates recovered from clinical samples obtained from inpatients at 16 hospitals in São Luis (Maranhão, Brazil). METHODS: The study was conducted using phenotypic tests and molecular methods, including polymerase chain reaction (PCR), sequencing and enterobacterial repetitive intergenic consensus (ERIC)-PCR. The nonparametric chi-square test of independence was used to evaluate the associations between the bacterial bla KPC gene and the modified Hodge test, and the chi-square adherence test was used to assess the frequency of carbapenemases and their association with the bla KPC gene. RESULTS: The most frequently isolated species were Acinetobacter baumannii (n = 128; 43.0%), K. pneumoniae (n = 75; 25.2%), and Pseudomonas aeruginosa (n = 42; 14.1%). Susceptibility assays showed that polymixin B was active against 89.3% of the bacterial isolates. The Acinetobacter spp. and K. pneumoniae strains were susceptible to amikacin and tigecycline, and Pseudomonas spp. were sensitive to gentamicin and amikacin. Among the 297 isolates, 100 (33.7%) were positive for the bla KPC gene, including non-fermentative bacteria (A. baumannii) and Enterobacteriaceae species. Among the isolates positive for the bla KPC gene, K. pneumoniae isolates had the highest positivity rate of 60.0%. The bla KPC gene variants detected included KPC-2, which was found in all isolates belonging to species of the Enterobacteriaceae family. KPC-2 and KPC-3 were observed in A. baumannii isolates. Importantly, the bla KPC gene was also detected in three Raoultella isolates and one isolate of the Pantoea genus. ERIC-PCR patterns showed a high level of genetic diversity among the bacterial isolates; it was capable of distinguishing 34 clones among 100 strains that were positive for bla KPC and were circulating in 11 of the surveyed hospitals. CONCLUSIONS: The high frequency of the bla KPC gene and the high degree of clonal diversity among microorganisms isolated from patients from different hospitals in São Luis suggest the need to improve the quality of health care to reduce the incidence of infections and the emergence of carbapenem resistance in these bacteria as well as other Gram-negative pathogens.

Epidemiologia do vírus do dengue em São Luís, Maranhão, no período de 2002 a 2012 / Epidemiology of dengue virus from 2002-2012 in Sao Luis, Maranhao state, Brazil

O presente estudo teve por objetivo relacionar a incidência do dengue com variáveis demográficas, temporais e meteorológicas, a fim de identificar as áreas com as mais elevadas incidências e determinar os sorotipos mais prevalentes, no período de 2002 a2012, no Município de São Luís, MA. Os dados foram extraídos dos boletins dos casos de dengue até então notificados, disponíveis na Secretaria Municipal de Saúde e Vigilância Epidemiológica e Sanitária do Município de São Luís, MA. Foi calculada estatística descritiva para todas as variáveis relevantes, como idade, sexo, condições climáticas, distribuição dos sorotipos e índice de infestação predial (IIP) pela larva de Aedes aegypti. Um total de 21.986 casos de dengue foi notificado ao Sistema de Vigilância Epidemiológica da Secretaria de Saúde e Vigilância Epidemiológica de São Luís, MA, o que correspondeu a 34,3% dos casos notificados no estado do Maranhão durante o período estudado. A faixa etária mais atingida foi a de 20 a49 anos, sem predomínio de sexo. A correlação entre os casos de dengue registrados e as condições meteorológicas, como pluviosidade, temperatura e umidade do ar, mostrou que a incidência de casos flutuou com essas variáveis climáticas. Assim, verificou-se ter ocorrido um aumento de casos de dengue durante o primeiro semestre dos anos estudados, que corresponde ao período chuvoso e de elevação de temperaturas, ao passo que, em intervalos de estiagem, a tendência foi de queda (meses de julho a dezembro). Nas mesmas condições, verificou-se um aumento da forma grave do dengue, como a febre hemorrágica do dengue, em população previamente exposta aos sorotipos 1, 2 e 3. O IIP pelo A. aegypti foi maior no período de2001 a 2007 e no ano de 2011. Nos anos em que foi registrada elevação dos índices de IIP, ocorreram os números mais elevados de casos de dengue no município de São Luis, MA.

Identification of Leptospira serovars by RFLP of the RNA polymerase beta subunit gene (rpoB).

Leptospires are usually classified by methods based on DNA-DNA hybridization and the conventional cross-agglutination absorption test, which uses polyclonal antibodies against lipopolysaccharides. In this study, the amplification of the rpoB gene, which encodes the beta-subunit of RNA polymerase, was used as an alternative tool to identify Leptospira. DNA extracts from sixty-eight serovars were obtained, and the hypervariable region located between 1990 and 2500-bp in the rpoB gene was amplified by polymerase chain reaction (PCR). The 600-bp amplicons of the rpoB gene were digested with the restriction endonucleases TaqI, Tru1I, Sau3AI and MslI, and the restriction fragments were separated by 6% polyacrylamide gel electrophoresis. Thirty-five fragment patters were obtained from the combined data of restriction fragment length polymorphism (PCR-RFLP) analysis and used to infer the phylogenetic relationships among the Leptospira species and serovars. The species assignments obtained were in full agreement with the established taxonomic classifications. Twenty-two serovars were effectively identified based on differences in their molecular profiles. However, the other 46 serovars remained clustered in groups that included more than one serovar of different species. This study demonstrates the value of RFLP analysis of PCR-amplified rpoB as an initial method for identifying Leptospira species and serovars.

Identification of Leptospira serovars by RFLP of the RNA polymerase beta subunit gene (rpoB)

Leptospires are usually classified by methods based on DNA-DNA hybridization and the conventional cross-agglutination absorption test, which uses polyclonal antibodies against lipopolysaccharides. In this study, the amplification of the rpoB gene, which encodes the beta-subunit of RNA polymerase, was used as an alternative tool to identify Leptospira. DNA extracts from sixty-eight serovars were obtained, and the hypervariable region located between 1990 and 2500-bp in the rpoB gene was amplified by polymerase chain reaction (PCR). The 600-bp amplicons of the rpoB gene were digested with the restriction endonucleases TaqI, Tru1I, Sau3AI and MslI, and the restriction fragments were separated by 6% polyacrylamide gel electrophoresis. Thirty-five fragment patters were obtained from the combined data of restriction fragment length polymorphism (PCR-RFLP) analysis and used to infer the phylogenetic relationships among the Leptospira species and serovars. The species assignments obtained were in full agreement with the established taxonomic classifications. Twenty-two serovars were effectively identified based on differences in their molecular profiles. However, the other 46 serovars remained clustered in groups that included more than one serovar of different species. This study demonstrates the value of RFLP analysis of PCR-amplified rpoB as an initial method for identifying Leptospira species and serovars.

Isolation of saint louis encephalitis virus from a horse with neurological disease in Brazil.

St. Louis encephalitis virus (SLEV) is a causative agent of encephalitis in humans in the Western hemisphere. SLEV is a positive-sense RNA virus that belongs to the Flavivirus genus, which includes West Nile encephalitis virus, Japanese encephalitis virus, Dengue virus and other medically important viruses. Recently, we isolated a SLEV strain from the brain of a horse with neurological signs in the countryside of Minas Gerais, Brazil. The SLEV isolation was confirmed by reverse-transcription RT-PCR and sequencing of the E protein gene. Virus identity was also confirmed by indirect immunofluorescence using commercial antibodies against SLEV. To characterize this newly isolated strain in vivo, serial passages in newborn mice were performed and led to hemorrhagic manifestations associated with recruitment of inflammatory cells into the central nervous system of newborns. In summary this is the first isolation of SLEV from a horse with neurological signs in Brazil.

Evaluation of laboratory tests for dengue diagnosis in clinical specimens from consecutive patients with suspected dengue in Belo Horizonte, Brazil.

BACKGROUND: Dengue is a widely spread arboviral disease in tropical and subtropical regions of the world. Dengue fever presents clinical characteristics similar to other febrile illness. Thus laboratory diagnosis is important for adequate management of the disease. OBJECTIVES: The present study was designed to evaluate the diagnostic performance of real-time PCR and serological methods for dengue in a real epidemic context. STUDY DESIGN: Clinical data and blood samples were collected from consecutive patients with suspected dengue who attended a primary health care unit in Belo Horizonte, Brazil. Serologic methods and real-time PCR were performed in serum samples to confirm dengue diagnosis. RESULTS: Among the 181 consecutive patients enrolled in this study with suspected dengue, 146 were considered positive by serological criteria (positive NS1 ELISA and/or anti-dengue IgM ELISA) and 138 were positive by real-time PCR. Clinical criteria were not sufficient for distinguishing between dengue and non-dengue febrile illness. The PCR reaction was pre-optimized using samples from patients with known viral infection. It had similar sensitivity compared to NS1 ELISA (88% and 89%, respectively). We also evaluated three commercial lateral flow immunochromatographic tests for NS1 detection (BIOEASY, BIORAD and PANBIO). All three tests showed high sensitivity (94%, 91% and 81%, respectively) for dengue diagnosis. CONCLUSION: According to our results it can be suggested that lateral flow tests for NS1 detection are the most feasible methods for early diagnosis of dengue.

Perfil epidemiológico dos marcadores sorológicos para vírus da hepatite B dos pacientes atendidos em um laboratório público / Epidemiological profile of serological markers for hepatitis B patients treated in a public laboratory

JUSTIFICATIVA E OBJETIVOS: A hepatite B é uma doença de distribuição universal que afeta ambos os sexos, podendo ser adquirida por meio de contato sexual, compartilhamento de seringas, exposição ocupacional e transfusão de sangue contaminado. O padrão de transmissão do vírus da hepatite B (VHB) está relacionado com a taxa de prevalência. O objetivo deste estudo foi determinar a prevalência de marcadores do VHB, de acordo com o índice de proteção vacinal. MÉTODO: A partir dos registros dos exames realizados em um laboratório público do município de São Luís, no ano de 2008, foram pesquisados os resultados para os marcadores de infecção com VHB: o anti-antígeno de superfície do VHB (HBsAg), os anticorpos anti-antígeno do core (anti-HBc) e anti-antígeno de superfície (anti-HBs).RESULTADOS: Dos 894 pacientes com sorologia positiva para VHB, 5,6% apresentaram marcador sorológico para fase aguda (HBsAg) prevalente em mulheres e pessoas com idade acima de 40 anos. Os anticorpos anti-HBc foram divididos em três tipos: anti-HBc (total), anti-HBc (IgG) e anti-HBc (IgM). Os índices de fase aguda e crônica utilizando estes marcadores foram similares (1,9% e 1,5%, respectivamente), com prevalência em mulheres e em pessoas com mais de 20 anos. O anti-antígeno de superfície anti-HBs foi detectado em 47,3% dos pacientes quando analisado isoladamente, indicando boa cobertura vacinal. CONCLUSÃO: Os dados do presente estudo indicam baixa prevalênciado VHB na população estudada.

BACKGROUND AND OBJECTIVES: Hepatitis B is a disease of worldwide distribution that affects both genders and can be acquired through sexual contact, needle sharing, occupational exposure and transfusion of contaminated blood. The pattern of transmission of hepatitis B virus (HBV) is related to the prevalence rate. The aim of this study was to determine the prevalence of HBV markers according to the rate of vaccine protection. METHOD: From the patient's records of a public laboratory of the city of São Luis (2008) the prevalence of three HBV infection markers, i.e., hepatitis B virus surface antigen (HBsAg), core anti-antigen (anti-HBc), and surface antigen antibody (anti-HBs) were studied. RESULTS: Of the 894 patients with positive serology for hepatitis B, 5.6% had prevalence of serologic marker for acute HBsAg in women and people over 40 years of age. The anti-HBc antibodies were divided into three types: anti-HBc (total), anti-HBc (IgG) and anti-HBc (IgM). The rates of acute and chronic use of these markers were similar (1.9% and 1.5% respectively) with the prevalence in women and in people over 20 years. The surface antigens anti-HBs were detected in 47.3% of patients when analyzed alone, indicating good vaccine coverage. CONCLUSION: The data from this study indicate low prevalence of hepatitis B virus among the studied population.