Effects of genetic polymorphisms on antioxidant status and concentrations of the metals in the blood of riverside Amazonian communities co-exposed to Hg and Pb.

There have been reports of genetic effects affecting the metabolism of Hg and Pb individually, and thus modulating their toxicities. However, there is still a knowledge gap with respect to how genetics may influence the toxicities of these toxic metals during a co-exposure scenario. This present study is therefore aimed at investigating the effects of polymorphisms in genes (GSTM1, GSTT1, GSTP1, GCLM, GCLC, GPx1, ALAD, VDR and MDR1) that have been implicated in Hg and Pb metabolisms affects the kinetics of these metals, as well as various blood antioxidant status parameters: MDA and GSH, and the activities of CAT, GPx and ALAD among populations that have been co-exposed to both Hg and Pb. Study subjects (207 men; 188 women) were from an Amazonian population in Brazil, exposed to Hg and Pb from diet. The blood levels of Hg and Pb were determined by ICP-MS while genotyping were performed by PCR assays. The median values of Hg and Pb in blood were 39.8µg/L and 11.0µg/dL, respectively. GSTM1, ALAD and VDR polymorphisms influenced Hg in blood (ß=0.17; 0.37 and 0.17; respectively, p<0.050) while variations on GCLM, GSTT1 and MDR1 (TT) modulated the concentrations of Pb among the subjects (ß=-0.14; 0.13 and -0.22; re-spectively, p<0.050). GSTT1 and GCLM polymorphisms also are associated to changes of MDA concentrations. Persons with null GSTM1 genotype had higher activity of the antioxidant enzyme CAT than carries of the allele. Individuals with deletion of both GSTM1 and GSTT1 had a decreased expression of GPx compared to those that expressed at least, one of the enzymes. ALAD 1/2 subjects had lower ALAD activity than individuals with the non-variant genotype. Our findings give further support that polymorphisms related to Hg and Pb metabolism may modulate Hg and Pb body burden and, consequently metals-induced toxicity.

Trichilia catigua against Helicobacter pylori: An in vitro, molecular and in silico approach.

Helicobacter pylori is a bacterium that is present in the stomach of about 50% of the global population and is associated with several gastric disorders, including cancer. Natural products with antimicrobial activity have been tested against H. pylori, among them Trichilia catigua (catuaba), which is widely distributed in Brazil. This study aimed to evaluate extracts of T. catigua bark against H. pylori via determination of the minimum inhibitory and bactericidal concentrations (MIC and MBC); evaluation of virulence factors by real-time PCR, synergism with standard antimicrobials and morphology by scanning electron microscopy and simulations of the mechanism of action by molecular docking. The ethyl acetate fraction provided the best results, with an MIC50 of 250 µg/mL and a 42.34% reduction in urease activity, along with reduced expression of the CagA and VacA genes, which encode for the main virulence factors. This fraction presented synergistic activity with clarithromycin, reducing the MIC of the drug by four-fold. Docking simulations suggested that the extracts inhibit fatty acid synthesis by the FAS-II system, causing damage to the cell membrane. Therefore, T. catigua extracts have potential as an adjuvant to treatment and are promising for the development of new anti-H. pylori drugs.

Modulation of gene expression and cell cycle by botryosphaeran, a (1→3)(1→6)-ß-d-glucan in human lymphocytes.

There is growing interest in the anticancer and immunomodulatory potential of fungal ß-d-glucans. In the present study, the modulation of gene expression via RT-qPCR and cell cycle kinetics via flow cytometry were assessed in human normal and tumor (Jurkat) lymphocytes after treatment with botryosphaeran (a fungal (1→3)(1→6)-ß-d-glucan) from Botryosphaeria rhodina MAMB-05. Cell cultures were treated with botryosphaeran either alone, or in combination with doxorubicin (DXR), in a post-treatment protocol. The expression of genes involved in immunomodulatory processes, apoptosis and cell cycle control, as well as ß-d-glucans cell receptors were assessed. Flow cytometry analysis identified tetraploid Jurkat cells in G1 phase when treated with botryosphaeran combined with DXR. This antiproliferative effect in G1 may be associated with down-regulation of the expression of genes involved in the G1 checkpoint. The repression of the CCR5 gene following botryosphaeran treatment, either alone or in combination with DXR, in tumor lymphocytes indicates a possible affinity of this particular (1→3)(1→6)-ß-d-glucan for the receptor CCR5. Therefore, botryosphaeran action appears to be involved in the repression of genes related to the G1 phase of the cell cycle and possibly in the interaction of the botryosphaeran, either alone, or in combination with DXR, with the CCR5 receptor.

Species distribution and in vitro fluconazole susceptibility of clinical Candida isolates in a Brazilian tertiary-care hospital over a 3-year period.

INTRODUCTION: In this study, we aimed at identifying Candida isolates obtained from blood, urine, tracheal secretion, and nail/skin lesions from cases attended at the Hospital Universitário de Londrina over a 3-year period and at evaluating fluconazole susceptibilities of the isolates. METHODS: Candida isolates were identified by polymerase chain reaction (PCR) using species-specific forward primers. The in vitro fluconazole susceptibility test was performed according to EUCAST-AFST reference procedure. RESULTS: Isolates were obtained from urine (53.4%), blood cultures (19.2%), tracheal secretion (17.8%), and nail/skin lesions (9.6%). When urine samples were considered, prevalence was similar in women (45.5%) and in men (54.5%) and was high in the age group >61 years than that in younger ones. For blood samples, prevalence was high in neonates (35%) and advanced ages (22.5%). For nail and skin samples, prevalence was higher in women (71.4%) than in men (28.6%). Candida albicans was the most frequently isolated in the hospital, but Candida species other than C. albicans accounted for 64% of isolates, including predominantly Candida tropicalis (33.2%) and Candida parapsilosis (19.2%). The trend for non-albicans Candida as the predominant species was noted from all clinical specimens, except from urine samples. All Candida isolates were considered susceptible in vitro to fluconazole with the exception of isolates belonging to the intrinsically less-susceptible species C. glabrata. CONCLUSIONS: Non-albicans Candida species were more frequently isolated in the hospital. Fluconazole resistance was a rare finding in our study.

[Hemolysis produced by Candida tropicalis isolates from clinical samples]. / Hemólise produzida por Candida tropicalis isoladas de amostras clínicas.

INTRODUCTION: Yeasts belonging to the genus Candida are responsible for the majority of fungal infections in humans. Candida tropicalis has been one of most commonly isolated non-albicans species. To analyze in vitro hemolysis promoted by clinical isolates of C. tropicalis obtained from blood and other clinical samples from hospitalized patients at the University Hospital of Londrina State University, Paraná, Brazil. METHODS: The hemolysis promoted by 28 clinical isolates of C. tropicalis was evaluated, and the isolates were grouped into classes according to the hemolysis levels. RESULTS: The majority of the blood isolates showed weak hemolysis (+), while the classes of strong hemolysis (+++) and very strong hemolysis (++++) predominated among isolates from other clinical samples such as urine, nail lesions and tracheal secretions. However, no statistical differences were detected (p> 0.05). CONCLUSIONS: Isolates of C. tropicalis obtained from different clinical samples showed a capacity to promote in vitro hemolysis.

Species distribution and in vitro fluconazole susceptibility of clinical Candida isolates in a Brazilian tertiary-care hospital over a 3-year period / Distribuição de espécies e suscetibilidade in vitro ao fluconazol de isolados clínicos de Candida em um hospital terciário brasileiro num período de três anos

INTRODUCTION: In this study, we aimed at identifying Candida isolates obtained from blood, urine, tracheal secretion, and nail/skin lesions from cases attended at the Hospital Universitário de Londrina over a 3-year period and at evaluating fluconazole susceptibilities of the isolates. METHODS: Candida isolates were identified by polymerase chain reaction (PCR) using species-specific forward primers. The in vitro fluconazole susceptibility test was performed according to EUCAST-AFST reference procedure. RESULTS: Isolates were obtained from urine (53.4 percent), blood cultures (19.2 percent), tracheal secretion (17.8 percent), and nail/skin lesions (9.6 percent). When urine samples were considered, prevalence was similar in women (45.5 percent) and in men (54.5 percent) and was high in the age group >61 years than that in younger ones. For blood samples, prevalence was high in neonates (35 percent) and advanced ages (22.5 percent). For nail and skin samples, prevalence was higher in women (71.4 percent) than in men (28.6 percent). Candida albicans was the most frequently isolated in the hospital, but Candida species other than C. albicans accounted for 64 percent of isolates, including predominantly Candida tropicalis (33.2 percent) and Candida parapsilosis (19.2 percent). The trend for non-albicans Candida as the predominant species was noted from all clinical specimens, except from urine samples. All Candida isolates were considered susceptible in vitro to fluconazole with the exception of isolates belonging to the intrinsically less-susceptible species C. glabrata. CONCLUSIONS: Non-albicans Candida species were more frequently isolated in the hospital. Fluconazole resistance was a rare finding in our study.

INTRODUÇÃO: Neste estudo objetivamos a identificação de isolados de Candida obtidos de sangue, urina, secreção traqueal e de lesões de unha/pele, de casos atendidos no Hospital Universitário de Londrina num período de três anos. Avaliamos também a suscetibilidade dos isolados ao fluconazol. MÉTODOS: Os isolados de Candida foram identificados pela reação em cadeia da polimerase (RCP) usando oligonucleotídeos iniciadores espécie-específicos. O teste de suscetibilidade in vitro ao fluconazol foi realizado segundo o procedimento de referência EUCAST-AFST. RESULTADOS: Isolados foram obtidos de urina (53,4 por cento), sangue (19,2 por cento), secreção traqueal (17,8 por cento) e lesões de unha/pele (9,6 por cento). Considerando as amostras de urina, a prevalência foi similar em mulheres (45,5 por cento) e em homens (54,5 por cento) e foi alta no grupo de idade > 61 anos do que em grupos mais jovens. Para amostras de sangue a prevalência foi alta em neonatos (35 por cento) e idades avançadas (22,5 por cento). Para amostras de unha e pele a prevalência foi maior em mulheres (71,4 por cento) do que em homens (28,6 por cento). Candida albicans foi a mais frequentemente isolada no hospital, mas outras espécies de Candida corresponderam a 64 por cento dos isolados, incluindo predominantemente Candida tropicalis (33,2 por cento) e Candida parapsilosis (19,2 por cento). A tendência de Candida não-albicans como espécie predominante foi observada para todas as amostras clínicas, exceto para amostras de urina. Todos isolados de Candida foram considerados suscetíveis, in vitro, ao fluconazol com exceção dos isolados pertencentes às espécies intrinsecamente menos suscetíveis C. glabrata. CONCLUSÕES: Espécies de Candida não-albicans foram mais frequentemente isoladas no hospital. Resistência ao fluconazol foi rara no nosso estudo.

Condições higiênico-sanitárias, qualidade microbiológica e teste de susceptibilidade antimicrobiana de cepas isoladas de sanduíches comercializados por ambulantes / Hygienic-sanitary condition, microbiological quality and antimicrobialsusceptibility of isolated strains from sandwiches sold in street markets

As condições higiênico-sanitárias e a qualidade microbiológica de locais de venda ambulantes que comercializam sanduíches em Londrina-PR foram investigadas neste estudo; as bactérias isoladas foram testados por meio de ensaios de sensibilidade a antimicrobianos. As condições higiênico-sanitárias foram analisadas em 47 pontos de venda de sanduíches tipo cachorro-quente e “cheese”, na cidade de Londrina, PR. Foram coletados aleatoriamente 20 sanduíches para análises microbiológicas, em que foi determinada a presença de bactérias mesófilas aeróbias estritas e facultativas, Staphylococcus aureus, coliformes a 35ºC e coliformes termotolerantes, Escherichia coli e Salmonella spp. Os micro-organismos isolados foram avaliados quanto à sensibilidade a antibióticos. As condições higiênico-sanitárias foram inadequadas em 79,2% dos itens avaliados. Todos os sanduíches estavam contaminados por bactérias mesófilas, coliformes a 35ºC e termotolerantes, e 45% por E. coli. S. aureus foi isolado de 55% das amostras, e em 25% os valores estavam acima do permitido pela legislação. Salmonella spp não foi detectada. A maioria dos isolados de E.coli e S. aureus foi resistente a pelo menos um tipo de antimicrobianos testados. A presença desses microorganismos pode estar associada com as condições higiênico-sanitárias inadequadas.

The hygienic-sanitary and microbiological qualities of sandwiches sold by street-vendors in Londrina-PRwere assessed, and the antimicrobial susceptibility testing was carried out on isolated bacteria. Sanitaryconditions were evaluated in 47 street-vending places, and 20 samples were randomly collected for analyzingthe contamination with Aerobic mesophilic bacteria, Staphylococcus aureus, total coliforms and coliformsat 45ºC, and Salmonella. The isolated microorganisms were identified and their sensitivity to antibioticswas evaluated. The sanitary conditions were inadequate in 79.2% of the evaluated items. All sandwiches(100%) were contaminated with mesophilic bacteria, coliforms at 35ºC and fecal coliform, and 45% withE. coli. S. aureus was found in 55% of samples, and in 25% these values were higher than those allowed bylegislation. No Salmonella sp was isolated from any of the tested samples. The hygiene practices of manyfood street-vending places are not adequate, resulting in a high proportion of read-to-eat foods withmicrobiological quality unsatisfactory for consumption.

Hemólise produzida por Candida tropicalis isoladas de amostras clínicas / Hemolysis produced by Candida tropicalis isolates from clinical samples

INTRODUÇÃO: Leveduras do gênero Candida são responsáveis pela maioria das infecções fúngicas em humanos. Candida tropicalis tem sido uma das mais comumente isoladas dentre as espécies não-albicans. O objetivo foi analisar a hemólise in vitro promovida por isolados clínicos de C. tropicalis provenientes de sangue e outras amostras clínicas de pacientes internados no Hospital Universitário da UEL, PR-Brasil. MÉTODOS: Foi avaliada a hemólise promovida por 28 isolados clínicos de C. tropicalis, sendo os isolados agrupados em classes de acordo com os níveis de hemólise. RESULTADOS: A maioria dos isolados de sangue apresentou hemólise fraca (+), enquanto as classes de hemólise forte (+++) e muito forte (++++) foram as predominantes nos isolados de outras amostras clínicas como urina, lesão de unha e secreção traqueal, embora não tenham sido detectadas diferenças estatísticas (p>0,05). CONCLUSÕES: Isolados de C. tropicalis, obtidos de diferentes amostras clínicas, apresentam capacidade de promover hemólise in vitro.

INTRODUCTION: Yeasts belonging to the genus Candida are responsible for the majority of fungal infections in humans. Candida tropicalis has been one of most commonly isolated non-albicans species. To analyze in vitro hemolysis promoted by clinical isolates of C. tropicalis obtained from blood and other clinical samples from hospitalized patients at the University Hospital of Londrina State University, Paraná, Brazil. METHODS: The hemolysis promoted by 28 clinical isolates of C. tropicalis was evaluated, and the isolates were grouped into classes according to the hemolysis levels. RESULTS: The majority of the blood isolates showed weak hemolysis (+), while the classes of strong hemolysis (+++) and very strong hemolysis (++++) predominated among isolates from other clinical samples such as urine, nail lesions and tracheal secretions. However, no statistical differences were detected (p> 0.05). CONCLUSIONS: Isolates of C. tropicalis obtained from different clinical samples showed a capacity to promote in vitro hemolysis.

Incidência e determinação do perfil de sensibilidade de cepas de Staphylococcus aureus isoladas da microbiota natural das fossas nasais e orofaringe de acadêmicos do curso de Enfermagem / Incidence and antimicrobial susceptibility of Staphylococcus aureus strains isolated from nasal and oral carriers, in student's nursing

Introdução - Staphylococcus aureus é um dos principais patógenos hospitalares e a resistência desses microrganismos contribui para a sua habilidade em disseminar-se entre os pacientes. A emergência e a disseminação de microrganismos resistentes aos antibióticos no âmbito hospitalar tem se tornado cada vez mais um problema grave e mundial para a Saúde Pública. O objetivo deste trabalho foi avaliar a freqüência de isolamento de S. aureus de mucosa nasal e oral de alunos do curso de Enfermagem, bem como determinar o perfil de sensibilidade deste microrganismo. Métodos - Foi utilizado swab, sendo friccionado nas mucosas nasal e oral de 55 alunos do 4º ano do curso de Enfermagem, e posterior inóculo em agar manitol salgado. As colônias suspeitas foram identificadas através de reação morfotintorial e dos testes de catalase, coagulase, acetoína e DNAse. Das 105 amostras obtidas de swab nasal e oral, 58% apresentaram-se positivas para presença de S. aureus, sendo que 20% apresentaram-se na região orofaríngea, 55% na mucosa nasal e 25% em ambos sítios anatômicos. Resultados - No teste de sensibilidade microbiana, 93,3% apresentaram sensibilidade aos antibióticos testados, 6,6% das amostras apresentaram resistência para sulfonamida, e 1,7% para eritromicina e cloranfenicol. Conclusão - Estes dados ressaltam a importância do estudo de distribuição de S.aureus, bem como determinar a sensibilidade/resistência a antibióticos, em profissionais da área de Saúde.

Introduction - Staphylococcus aureus is a very important hospital pathogen and its resistence to antibiotics is associates to several nosocomial infection. The emergence and dissemination of resistent microrganisms have became a serious problem to the public health. This study aimed the analysis of S. aureus from nasal and oral cavities of nurse´s students and the sensitive profile of the isolates to different antimicrobial agents. Methods - It were utilized samples of nasal and oral flora from 55 individuals were swabbed with a sterile swab and inoculated on mannitol salt agar. All samples were identified using biochemical characteristics for the species identification,i.e. Gram, calatase, coagulase and DNAse. S. aureus was found in 58% of the carriers, being 20% of oral carriers, 55% of nasal carriers and 25% from nasal and oral carriers. Results - Some 93.3% of all 105 strains showed sensitivity to the antimicrobial agents; 6.6% showed resistence to sulfonamide, and 1.7% showed resistence to erithromicin and chloramphenicol agents. Conclusion - This dates were important for our purpose, showed the distribution of S.aureus and the correlation with sensibility and resistance to drugs in mucosa carriers student's nursing.

Incidência e determinação do perfil de sensibilidade de cepas de Staphylococcus aureus isolada da microbiota natural das fossas nasais e orofaringe de acadêmicos do curso de Enfermagem / Incidence and antimicrobial susceptibility of Staphylococcus aureus strains isolated from nasal and oral carriers, in student's nursing

Introdução - Staphylococcus aureus é um dos principais patógenos hospitalares e a residência desses microorganismos contribui para a sua habilidade em disseminar-se entre os pacientes. A emergência e a disseminação de microorganismos resistentes aos antibióticos no âmbito hospitalar tem se tornado cada vez mais um problema grave e mundial para a Saúde Pública. O objetivo desse trabalho foi avaliar a freqüência de isolamento de S. aureus de mucosa nasal e oral de alunos do curso de Enfermagem, bem como determinar o perfil de sensibilidade deste microorganismo. Métodos - Foi utilizado swab, sendo friccionado nas mucosas nasal e oral de 55 alunos do 4º ano do curso de Enfermagem, e posterior inóculo em agar manitol salgado. As colônias suspeitas foram identificadas através de reação morfotintorial e dos testes de catalase, coagulase, acetoína e DNAse. Das 105 amostras obtidas de swab nasal e oral, 58% apresentaram-se positivas para a presença de S. aureus, sendo que 20% apresentaram-se na região orofaríngea, 55% na mucosa nasal e 25% em ambos os sítios anatômicos. Resultados - No teste de sensibilidade microbiana, 93,3% apresentaram sensibilidade aos antibióticos testados, 6,6% das amostras apresentaram resistência para sulfonamida, e 1,7% para eritromicina e cloranfenicol. Conclusão - Estes dados ressaltam a importância do estudo de distribuição de S. aureus, bem como determinar a sensibilidade /resistência a antibióticos, em profissionais da área de saúde.