A haplotype in the dipeptidyl peptidase 4 gene impacts glycemic-related traits of Brazilian older adults

Dipeptidyl peptidase 4 (DPP4) regulates various physiological pathways and has a pivotal role in glucose homeostasis. The objective of this study was to verify the association of a haplotype constituted by two single nucleotide polymorphisms (rs2268894 and rs6741949) in the DPP4 gene with type 2 diabetes mellitus (T2DM) and fasting glycemia-related variables in a sample of Brazilian older adults, taking serum levels and enzymatic activity of DPP4 into account. Clinical, biochemical, and anthropometric characteristics as well as DPP4 serum levels and enzymatic activity were determined in 800 elderly (≥60 years old) individuals. Assessment of polymorphic sites was performed by real-time PCR whereas haplotypes were inferred from genotypic frequencies. Statistical analyses compared measures and proportions according to T2DM diagnosis and DPP4 haplotypic groups. The most common haplotype consisted of the T-rs2268894/G-rs6741949 string, which was 20% more frequent among non-diabetics. Considering non-diabetic patients alone, carriers of the T/G haplotype had significantly lower levels of blood glucose, insulin, HOMA-IR index, and DPP4 activity. Among diabetic patients, the T/G haplotype was associated with lower DPP4 levels whereas glycemic scores were not affected by allelic variants. Our results suggested that the genetic architecture of DPP4 affects the glycemic profile and DPP4 serum levels and activity among elderly individuals according to the presence or absence of T2DM, with a possible implication of the T/G haplotype to the risk of T2DM onset.

Standardization of a rapid quadruplex PCR method for the simultaneous detection of bovine, buffalo, Salmonella spp, and Listeria monocytogenes DNA in milk / [Padronização de um método rápido de PCR quadriplex para a detecção simultânea de DNA de bovino, bubalino, Salmonella spp. e Listeria monocytogenes em leite]

The objective of the present study was to Standardize a Polymerase Chain Reaction (PCR) protocol for the authentication of bovine and buffalo milk, and to detect the presence of Salmonella spp. and Listeria monocytogenes. For this, the target DNA was extracted, mixed, and subjected to a PCR assay. Milk samples were defrauded and experimentally contaminated with microorganisms to assess the detection of target DNA at different times of cultivation, bacterial titers, and concentration of genetic material. In addition, the protocol was tested with DNA extracted directly from food, without a pre-enrichment step. The proposed quadruplex PCR showed good accuracy in identifying target DNA sequences. It was possible to simultaneously identify all DNA sequences at the time of inoculation (0h), when the samples were contaminated with 2 CFU/250mL and with 6h of culture when the initial inoculum was 1 CFU/250mL. It was also possible to directly detect DNA sequences from the food when it was inoculated with 3 CFU/mL bacteria. Thus, the proposed methodology showed satisfactory performance, optimization of the analysis time, and a potential for the detection of microorganisms at low titers, which can be used for the detection of fraud and contamination.

O objetivo do presente estudo foi padronizar um protocolo de reação em cadeia da polimerase (PCR) para a autenticação de leite bovino e bubalino e a detecção da presença de Salmonella spp. e Listeria monocytogenes. Para isso, o DNA-alvo foi extraído, misturado e submetido ao ensaio de PCR. Amostras de leite foram fraudadas e contaminadas experimentalmente com os micro-organismos, para se avaliar a detecção do DNA-alvo em diferentes tempos de cultivo, os títulos bacterianos e a concentração de material genético. Além disso, o protocolo foi testado com DNA extraído diretamente do alimento, sem a etapa de pré-enriquecimento. A PCR quadriplex proposta mostrou boa precisão na identificação de sequências de DNA-alvo. Foi possível identificar simultaneamente todas as sequências de DNA no momento da inoculação (0h), quando as amostras estavam contaminadas com 2 UFC/250mL, e com seis horas de cultura, quando o inóculo inicial foi de 1 UFC/250mL. Também foi possível detectar diretamente as sequências de DNA do alimento quando este foi inoculado com 3 UFC/mL de bactérias. Dessa forma, a metodologia proposta apresentou desempenho satisfatório, otimização do tempo de análise e potencial para detecção de micro-organismos em baixos títulos, podendo ser utilizada para detecção de fraude e contaminação.

Monitoring of Leptospira in captive turtles by DNA analysis / [Monitoramento de Leptospira em quelônios mantidos em cativeiro pela análise de DNA]

Existem poucos estudos sobre doenças infecciosas em animais silvestres. O objetivo deste estudo foi pesquisar DNA de Leptospira spp. em sangue de tartarugas mantidas em cativeiro, pertencentes ao Bosque Rodrigues Alves (Jardim Zoobotânico da Amazônia). O DNA foi isolado das amostras de sangue coletadas de 148 tartarugas pertencentes a seis espécies diferentes. A reação em cadeia da polimerase (PCR) foi realizada utilizando-se iniciadores específicos para DNA de Leptospira spp. Nenhuma das amostras apresentou resultado positivo para Leptospira spp.(AU)

The APOB rs693 polymorphism impacts the lipid profile of Brazilian older adults

The apolipoprotein B (APOB) gene contains several polymorphic sites described as risk modifiers for cardiovascular events. The objective of this study was to verify the association of the classic APOB Xba I polymorphism (rs693) with atherosclerotic risk factors in a segment of the Brazilian elderly population considering their usual dietary intake. Clinical and biochemical characteristics as well as total caloric and fat intake data were determined from 644 elderly individuals. Polymorphism analysis was performed by conventional polymerase chain reaction followed by enzyme restriction. Statistical analyses compared measures and proportions according to different APOB genotypic combinations. Statistically significant association was found between Xba I polymorphism and serum LDL, total cholesterol, and total lipid levels, with important elevations among T homozygotes compared to the other genotypes. There was homogeneity in all other parameters analyzed (including intake pattern), with a tendency for reduced levels of circulating apolipoprotein B among TT individuals. Our results pointed out that genetic variation in APOB affected the lipemic profile of elderly individuals in a context not biased by diet, generating a pattern suggestive of secretory disorder of lipoprotein particles, with possible implication in atherosclerotic risk.

Padronização de uma PCR para a autenticação do Salmo salar em pratos da culinária japonesa / Standardization of a PCR for the authentication of Salmo salar in dishes of Japanese cuisine

O objetivo deste trabalho foi padronizar uma PCR para a detecção do Salmo salar, a qual possa ser usada na autenticação do salmão utilizado em pratos da culinária japonesa e do pescado comercializado in natura. Para isso, dois lotes de sushi foram produzidos experimentalmente. Além disso, foram visitados 38 estabelecimentos que comercializam comida japonesa e 10 peixarias na região metropolitana de Belém, visando à coleta do sushi, do temaki e do pescado pertencente à espécie Salmo salar. Os dados demonstraram que a técnica foi eficiente para a autenticação de Salmo salar, visto que a espécie foi detectada tanto nas amostras de sushis preparados experimentalmente quanto nas alíquotas de pescados isolados, utilizados para a preparação do sushi. Em contrapartida, a espécie Salmo trutta não foi detectada nas amostras de sushis preparados com esta espécie nem nas alíquotas de pescado isolado. Além disso, foi possível a confirmação da utilização da espécie Salmo salar no preparo das amostras de sushi, temaki e de pescado. Portanto, concluiu-se que a técnica foi capaz de amplificar o DNA da referida espécie e não gerou identificação inespecífica quando a espécie Salmo trutta foi analisada, podendo ser uma ferramenta adequada para a autenticação do Salmo salar.(AU)

The objective of this work was to standardize a PCR for the detection of Salmo salar, which can be used in the authentication of salmon used in Japanese dishes and fish commercialized in natura. For this, two batches of sushi were produced experimentally. In addition, 38 establishments that sell Japanese food and 10 fishmongers in the metropolitan area of Belém were visited, aiming to collect sushi, temaki and fish belonging to the species Salmo salar. The data demonstrated that the technique was efficient for the authentication of Salmo salar, since the species was detected in both the experimentally prepared sushi samples and the isolated fish aliquots used for the preparation of sushi. In contrast, the species Salmo trutta was not detected in the sushi samples prepared with this species nor in the isolated fish aliquots. In addition, it was possible to confirm the use of the Salmo salar species in the preparation of sushi, temaki and fish samples. Therefore, it was concluded that the technique was able to amplify the DNA of this species and did not generate nonspecific identification when the species Salmo trutta was analyzed, being able to be a suitable tool for the authentication of Salmo salar.(AU)

Imunogenicidade da proteína M recombinante de Streptococcus equi subsp. equi coadministrada com um adjuvante molecular / Immunogenicity of recombinant M protein of Streptococcus equi subsp. equi co administered with a molecular adjuvant

The strangles is an infectious disease that affects horses from all ages and causes important economic losses in the equine-related business. The aim of this work was to evaluate the immunogenicity of the recombinant M protein from Streptococcus equi (rSeM) co-administered with the recombinant heat-labile enterotoxin B subunit from Escherichia coli (rLTB) in mice and horses. A total of 72 female Balb-c mice were divided into eight groups and 18 horses were divided into six groups. The animals were inoculated by intramuscular (IM) or intranasal (IN) routes with different treatments of rSeM, rLTB and/or Al(OH)3. The results obtained in both species, independent of administration routes, demonstrated that rSeM + rLTB had higher levels of specific serum immunoglobulins, however, in mucosal immunity the increase was not identified. Thus, the use of rSeM as vaccine antigen and rLTB as adjuvant can be a potential tool in the control of equine strangles.(AU)

Superação de dormência em sementes de Bombax malabaricum D. C. (Malvaceae) / Dormancy breaking in seeds of Bombax malabaricum D. C. (Malvaceae)

RESUMOA espécie Bombax malabaricum, popularmente conhecida por paineira-vermelha, apresenta metabólitos secundários de interesse medicinal. Como outras espécies da família Malvaceae, suas sementes possuem dormência tegumentar, o que dificulta sua propagação sexuada. Assim, o presente trabalho objetivou superar a dormência de sementes de B.malabaricum por métodos físicos e químicos. Os métodos utilizados na escarificação mecânica foram: abrasão do tegumento com lixa e corte com tesoura na região oposta ao hilo. Para a escarificação química foram utilizados: ácido clorídrico (HCl), ácido nítrico (HNO3), ácido sulfúrico (H2SO4), hidróxido de sódio (NaOH) e hidróxido de potássio (KOH) por um período de 30 minutos. Também houve imersão de sementes em 40 mg L-1 de GA3 pelo mesmo período. Foram utilizadas quatro repetições por tratamento, constituídas de 25 sementes por placa e conduzidas em Câmara de Germinação. Os dados obtidos foram utilizados para o cálculo da Germinabilidade, Índice de Velocidade de Germinação, Tempo Médio de Germinação, e Porcentagem de Sementes Deterioradas. O HNO3 mostrou-se como o melhor tratamento na variável Germinabilidade (83,5%), seguido do HCl (71,5%) e das técnicas de escarificação mecânica, com lixa (71,5%) e tesoura (67,5%). O HNO3também apresentou o menor número de sementes deterioradas (3,5%); o H2SO4 foi o tratamento que mais causou danos às sementes (36,5%).

ABSTRACTThe species Bombax malabaricum, popularly known as red-cotton tree, presents secondary metabolites of medicinal interest. Like other species of the Malvaceae, their seeds have tegumentary dormancy, which hinders their sexual propagation. Thus, this study aimed to break the dormancy of B. malabaricum through physical and chemical strategies. For mechanical scarification of the seeds it were employed: abrasion of the tegument with sandpaper and cut with scissors in the region opposite to the hilum. For chemical scarification: hydrochloric acid (HCl), nitric acid (HNO3), sulphuric acid (H2SO4), sodium hydroxide (NaOH) and potassium hydroxide (KOH) for a period of 30 minutes. The seeds were also subjected to immersion in 40 mg L-1 of GA3 for the same period. Four replicates of 25 seeds were used for each treatment and then they were conducted for calculation in a B.O.D. germination Chamber. The results obtained were applied in order to calculate: Germination, Germination Speed Index, Average Time of Germination and Percentage of Damaged Seeds. The HNO3proved to be the best treatment in the Germination trials (83.5%), followed by the HCl (71.5%) and the mechanical scarification techniques with sandpaper (71.5%) and scissors (67.5%). The HNO3 also showed the lowest percentage of damaged seeds (3.5%); The H2SO4 was the treatment that caused more damage to the seeds (36.5%).

Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA / Clonagem, expressão e caracterização de proteína SeM de Streptococcus equi subsp. equi e avaliação de sua utilização como antígeno em um ELISA indireto

Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd...

A adenite equina é uma enfermidade economicamente importante de equinos, causada por Streptococcus equi subsp. equi. Seu diagnóstico pode ser confirmado de forma direta, por meio de isolamento bacteriano e de PCR, ou de forma indireta, por meio de ELISA, método baseado na detecção de anticorpos séricos. O objetivo deste estudo foi clonar, expressar e caracterizar a proteína SeM de Streptococcus equi subsp. equi, avaliar sua utilização como antígeno em um ELISA indireto e determinar a capacidade do teste de distinguir soros de animais negativos, vacinados e positivos. Para tal, foi inicialmente realizada a clonagem do gene que codifica para a proteína SeM e sua expressão em Escherichia coli. Posteriormente, a proteína produzida foi caracterizada e utilizada como antígeno em um teste de ELISA indireto. Para avaliação do teste, foram utilizadas amostras de soro de 40 potros negativos, de 46 equinos vacinados com uma vacina comercial contra adenite equina e de 46 equinos com diagnóstico da doença. O teste demonstrou alta sensibilidade e especificidade, permitindo discriminar entre soros negativos e positivos, positivos e de animais vacinados, e negativos e de animais vacinados. Assim, conclui-se que a proteína rSeM produzida pode ser usada como antígeno para o diagnóstico da enfermidade e que o ELISA descrito pode ser útil para avaliar o estado imunológico do rebanho...

Testes fitoquímicos em extratos foliares de Oeceoclades maculata Lindl. (Orchidaceae) / Phytochemical tests in Oeceoclades maculata Lindl. (Orchidaceae) leaf extracts

O presente trabalho teve por objetivo determinar a capacidade antioxidante de Oeceoclades maculata. A atividade foi determinada de acordo com a capacidade do extrato hidroalcoólico das folhas reduzir o radical DPPH. Os resultados obtidos permitem concluir que os conteúdos de polifenóis e flavonas/flavonóis são maiores no extrato preparado a frio (EBHF) do que no extrato preparado a quente (EBHQ), sugerindo que o aquecimento usado na preparação do extrato altera esses componentes ativos. Portanto, para esta espécie deve ser realizado o método de extração a frio para conservar as substâncias que possuem atividade antioxidante.

This work aimed to determine the antioxidant capacity of Oeceoclades maculata. The antioxidant activity was determined in accordance with the ability of the hydroalcoholic extract of the leaves to reduce the DPPH radical. The results showed that the polyphenol and flavonoid contents are greater in the cold extract (EBHF) than in the hot extract (EBHQ), which suggests that the heating effect in the preparation of the extract modifies these active compounds. Therefore, for this species, the cold extraction method must be carried in order to conserve the substances with antioxidant activity.

Outcomes and organ dysfunctions of critically ill patients with systemic lupus erythematosus and other systemic rheumatic diseases

Our objective was to compare the pattern of organ dysfunctions and outcomes of critically ill patients with systemic lupus erythematosus (SLE) with patients with other systemic rheumatic diseases (SRD). We studied 116 critically ill SRD patients, 59 SLE and 57 other-SRD patients. The SLE group was younger and included more women. Respiratory failure (61 percent) and shock (39 percent) were the most common causes of ICU admission for other-SRD and SLE groups, respectively. ICU length-of-stay was similar for the two groups. The 60-day survival adjusted for the groups’ baseline imbalances was not different (P = 0.792). Total SOFA scores were equal for the two groups at admission and during ICU stay, although respiratory function was worse in the other-SRD group at admission and renal and hematological functions were worse in the SLE group at admission. The incidence of severe respiratory dysfunction (respiratory SOFA >2) at admission was higher in the other-SRD group, whereas severe hematological dysfunction (hematological SOFA >2) during ICU stay was higher in the SLE group. SLE patients were younger and displayed a decreased incidence of respiratory failure compared to patients with other-SRDs. However, the incidences of renal and hematological failure and the presence of shock at admission were higher in the SLE group. The 60-day survival rates were similar.

Morfologia e histologia do oviduto de marrecas Ana boschas / Morphology and histology of Ana Boschasfemale ducks oviduct

Avaliaram-se o comprimento do infundíbulo, do magno, do istmo, do útero e da vagina e o número de pregas do magno e do istmo do oviduto de 20 marrecas Ana boschas na fase reprodutiva. O infundíbulo apresenta mucosa com pregas longitudinais e baixas, revestidas por epitélio pseudoestratificado cilíndrico ciliado, com células caliciformes. O magno, compartimento mais longo do oviduto, 25,38cm±3,20, encontra-se constituído por uma camada mucosa com pregas altas e espessas revestidas por células cilíndricas ciliadas e abundantes células caliciformes. O istmo é formado por uma mucosa com pregas estreitas e curtas e numerosas glândulas tubulares que se estendem para o interior da lâmina própria. O útero, região curta do oviduto, 5,25cm±1,26, apresenta parede com pregas e cristas baixas e numerosas glândulas tubulares enoveladas, dirigidas para o interior da lâmina própria. A vagina, um estreito tubo muscular, está constituído por oito anéis circulares, em média, e uma camada muscular altamente desenvolvida e espessa. A morfologia do oviduto da marreca apresenta características morfológicas e histológicas distintas dos galiniformes, observando-se que a vagina e a porção cranial do infundíbulo apresentam pregas e células caliciformes, respectivamente, sendo estas últimas estruturas ausentes nos galiniformes.

The length of infundibulum, magnum, isthmus, uterus, vagina, and the number of oviduct, magnum, and isthmus folds were evaluated in 20 Ana boschas female ducks in the reproductive phase. The infundibulum presented mucous membrane with longitudinal and short folds, covered by ciliated columnar pseudostratified epithelium, with goblet cells. Magnum, the longest oviduct compartment, 25.38cm±3.20, is constituted by mucous membrane with high and thick folds that are covered by ciliated columnar cells and many goblet cells. Isthmus is formed by mucous membrane with narrow and short folds, and many tubular glands that extend inside lamina propria. Uterus, an oviduct short region, 5.25cm±1.26, presented surface with short folds and crests, and numerous reeled tubular glands, that are directed inside lamina propria. Vagina, a strait muscular tube, is constituted by, approximately, eight circular rings, and a very developed and thick muscular layer. The oviduct morphology of female ducks presented different morphological and histological characteristics from Galliformes, because vagina and infundibulum cranial portion present folds and goblet cells, respectively, and the last cited structures are absent in Galliformes.

Imaturos de Sarsina violascens (Herrich-Schäffer) (Lepidoptera, Noctuidae, Lymantriinae) / Immatures of Sarsina violascens (Herrich-Schäffer) (Lepidoptera, Noctuidae, Lymantriinae)

Imaturos de Sarsina violascens (Herrich-Schäffer) (Lepidoptera, Noctuidae, Lymantriinae). Sarsina violascens é uma espécie polífaga que eventualmente se alimenta de P. cattleianum Sabine (Myrtaceae) durante sua fase larval. Neste estudo são descritas a morfologia e o comportamento dos imaturos, com ilustrações, fotografias e imagens de microscopia eletrônica de varredura.

Sarsina violascens is a polyphagous species that also feeds on P. cattleianum Sabine (Myrtaceae) during the larval stages. In this study the morphology and the behaviour of the immature stages of this species are described and illustrated, including scanning electronic microscopy photographs.

Acute exercise performed close to the anaerobic threshold improves cognitive performance in elderly females

The objective of the present study was to compare the effect of acute exercise performed at different intensities in relation to the anaerobic threshold (AT) on abilities requiring control of executive functions or alertness in physically active elderly females. Forty-eight physically active elderly females (63.8 ± 4.6 years old) were assigned to one of four groups by drawing lots: control group without exercise or trial groups with exercise performed at 60, 90, or 110 percent of AT (watts) and submitted to 5 cognitive tests before and after exercise. Following cognitive pretesting, an incremental cycle ergometer test was conducted to determine AT using a fixed blood lactate concentration of 3.5 mmol/L as cutoff. Acute exercise executed at 90 percent of AT resulted in significant (P < 0.05, ANOVA) improvement in the performance of executive functions when compared to control in 3 of 5 tests (verbal fluency, Tower of Hanoi test (number of movements), and Trail Making test B). Exercising at 60 percent of AT did not improve results of any tests for executive functions, whereas exercise executed at 110 percent of AT only improved the performance in one of these tests (verbal fluency) compared to control. Women from all trial groups exhibited a remarkable reduction in the Simple Response Time (alertness) test (P = 0.001). Thus, physical exercise performed close to AT is more effective to improve cognitive processing of older women even if conducted acutely, and using a customized exercise prescription based on the anaerobic threshold should optimize the beneficial effects.

ALT-C, a disintegrin-like Cys-rich protein from Bothrops alternatus, increases skeletal myoblast viability

ALT-C, an ECD motif (glutamic acid, cysteine, aspartic acid) disintegrin from Bothrops alternatus snake venom, induces alfa2beta1 integrin-mediated signaling and neutrophil chemotaxis. In vitro, in human umbilical vein endothelial cells (HUVEC), ALT-C induces cell proliferation, thus showing an interesting potential for tissue regeneration studies. This work aimed to evaluate the influence of ALT-C in myoblast viability and differentiation. Myoblasts were obtained from hind limb muscles of 3 to 4-day old Wistar rats. The cells were incubated with ALT-C at different concentrations and incubation periods were followed by total RNA isolation. cDNA synthesis and real time polymerase chain reaction (PCR) were performed with primers of myoD as well as of both (slow and fast) myosin heavy chain isoforms (MHC). ECD-disintegrin increased myoblast viability in a dose-dependent way, mostly with 50 to 100 nM concentrations, and such effect was more prevalent after 48 hours. No changes in gene expression of both MHC isoforms were observed in ALT-C-treated cells. MyoD expression was not detected, which suggests that myoblasts were in mature stages. Protease activity and cytokine array tested in a medium of 50 nM ALT-C-treated cells after 48 hours were not different from controls. In conclusion, it was shown that myoblats are sensitive to ALT-C indicating an integrin-mediated intracellular signaling that increases cell viability.

Resistência a antimicrobianos dependente do sistema de efluxo multidrogas em Escherichia coli isoladas de leite mastítico / Antimicrobial resistance dependent on multidrugs efflux in Escherichia coli isolated from the mastitic milk

Identificaram-se e caracterizaram-se a resistência e a multirresistência aos principais antimicrobianos usados no tratamento de mastite bovina causada por Escherichia coli. A concentração inibitória mínima (MIC) e o sistema de efluxo foram detectados pelas curvas de crescimento, com base na densidade óptica, em diferentes concentrações da droga e na presença e na ausência do desacoplador da força próton-motora (PMF). E. coli 1 foi resistente à neomicina e à gentamicina; E. coli 3 e 4, à tetraciclina e à estreptomicina; e E. coli 2 e 6 à gentamicina. E. coli 5 apresentou modelo de sensibilidade. Observou-se que MICs de todos os antimicrobianos dos multirresistentes (E. coli 1, 3 e 4) diminuíram na presença do desacoplador, o que sugere sistema de efluxo multidrogas. Após cura, apenas E. coli 1 apresentou modelo de sensibilidade, porém não houve alterações das MICs, antes e após adição do desacoplador. Os resultados indicam possível presença de mecanismo de resistência dependente da PMF codificado, ou parte dele, em plasmídeo.

Resistance and multiresistance to main antimicrobials used for treating bovine mastitis caused by Escherichia coli were identified and characterized. The minimal inhibitory concentration (MIC) and efflux systems were detected by the use of growth curves based on optical density at different drug concentrations and both presence and absence of uncoupler of the proton-motive force (PMF). E. coli 1 was resistant to neomycin and gentamycin, E. coli 3 and 4 were resistant to tetracycline and streptomycin, whereas E. coli 2 and 6 were resistant to gentamycin. E. coli 5 showed sensibility model. MICs of all antimicrobials of the multiresistant samples (E. coli 1, 3, and 4) were decreased in presence of the uncoupler, therefore suggesting the presence of the multidrug efflux system. After healing, only E. coli 1 showed sensibility model, however no alteration occurred in MIC(s) before and after adding the uncoupler. Those data inform the possible presence of a PMF dependent resistance mechanism that is totally or partly codified in plasmid.

Identification of Lactobacillus UFV H2b20 (probiotic strain) using DNA-DNA hybridization / Identificação de Lactobacillus UFV H2b20 (linhagem probiótica) através de hibridização DNA-DNA

Sequence analyses of the 16S rDNA gene and DNA-DNA hybridization tests were performed for identification of the species of the probiotic Lactobacillus UFV H2b20 strain. Using these two tests, we concluded that this strain, originally considered Lact. acidophilus, should be classified as Lact. delbrueckii.

Análise da seqüência do gene 16S rDNA e ensaios de hibridização DNA_DNA foram empregados para identificar a espécie da cepa probiótica Lactobacillus UFV H2b20. Empregando-se estes dois testes, concluímos que esta cepa, originalmente considerada Lact. acidophilus, deve ser classificada como Lact. delbrueckii.

Identification of Lactobacillus UFV H2b20 (probiotic strain) using DNA-DNA hybridization

Sequence analyses of the 16S rDNA gene and DNA-DNA hybridization tests were performed for identification of the species of the probiotic Lactobacillus UFV H2b20 strain. Using these two tests, we concluded that this strain, originally considered Lact. acidophilus, should be classified as Lact. delbrueckii.

Análise da seqüência do gene 16S rDNA e ensaios de hibridização DNA_DNA foram empregados para identificar a espécie da cepa probiótica Lactobacillus UFV H2b20. Empregando-se estes dois testes, concluímos que esta cepa, originalmente considerada Lact. acidophilus, deve ser classificada como Lact. delbrueckii.

Association between the -174 G/C promoter polymorphism of the interleukin-6 gene and cardiovascular disease risk factors in Brazilian older women

In worldwide studies, interleukin-6 (IL-6) is implicated in age-related disturbances. The aim of the present report was to determine the possible association of IL-6 -174 C/G promoter polymorphism with the cytokine profile as well as with the presence of selected cardiovascular risk features. This was a cross-sectional study on Brazilian women aged 60 years or older. A sample of 193 subjects was investigated for impaired glucose regulation, diabetes, hypertension, and dyslipidemia. Genotyping was done by direct sequencing of PCR products. IL-6 and C-reactive protein were quantified by high-sensitivity assays. General linear regression models or the Student t-test were used to compare continuous variables among genotypes, followed by adjustments for confounding variables. The chi-square test was used to compare categorical variables. The genotypes were consistent with Hardy-Weinberg equilibrium proportions. In a recessive model, mean waist-to-hip ratio, serum glycated hemoglobin and serum glucose were markedly lower in C homozygotes (P = 0.001, 0.028, and 0.047, respectively). In a dominant hypothesis, G homozygotes displayed a trend towards higher levels of circulating IL-6 (P = 0.092). Non-parametric analysis revealed that impaired fasting glucose and hypertension were findings approximately 2-fold more frequent among G homozygous subjects (P = 0.042 and 0.043, respectively). Taken together, our results show that the IL-6 -174 G-allele is implicated in a greater cardiovascular risk. To our knowledge, this is the first investigation of IL-6 promoter variants and age-related disturbances in the Brazilian elderly population.

rACLF, a recombinant snake venom metalloprotease, activates endothelial cells in vitro

Snake venom metalloproteases (SVMPs) comprise a family of snake venom toxins responsible for most of local and systemic effects observed during envenomation by snakes from the Viperidae family. The vascular system and more specifically the endothelium seem to be the preferential targets of these proteins. This work describes the effects of rACLF, a recombinant SVMP from Agkistrodon contortrix laticinctus on human umbilical vein endothelial cells (HUVECs) in vitro. Our results showed that rACLF activates HUVECs by the release of mediators involved in inflammation and hemostasis such as prostacyclin and interleukin-8. We also demonstrated that rACLF increased the expression of ICAM-I and decay accelerating factor (DAF). Moreover, rACLF protects the HUVECs against apoptosis induced by serum deprivation. These results suggest that the endothelial cell activation induced by SVMPs may have a significant role in the development of the local inflammatory lesion observed in Viperidae envenomation.

Preparação e caracterização inicial de complexo de inclusão entre nitrofurazona e 2-hidroxipropil-Beta-ciclodextrina / Preparation and initial characterization of inclusion complex between nitrofurazone and 2-hydroxypropyl-Beta-cyclodextrin

Nitrofurazona(NF), 5-nitro-2-furaldeído semicarbazona, é um antibiótico de amplo espectro, que apresenta diversos efeitos tóxicos e baixa solubilidade aquosa. A complexação da NF com ciclodextrinas é de grande interesse para o desenvolvimento de uma formulação para este antibiótico que seja mais segura e eficiente. Neste trabalho foi realizada a preparação e caracterização inicial do complexo de inclusão entre NF e hidroxipropil-Beta-ciclodextrina (HP-Beta-CD) através de experimentos para determinação da cinética de complexação, medidas de fotoestabilidade, medidas de constante de afinidade fármaco: ciclodextrina, ensaios de liberação in vitro, estequiometria de formação do complexo e morfologia do complexo por microscopia eletrônica de varredura. Os ensaios de cinética de complexação mostram que para o complexo atingir o equilíbrio são necessárias 17,3h. As isotermas de solubilidade determinadas para a NF em função da temperatura mostraram perfis do tipo A e B indicando que a temperatura é um fator importante na complexação da NF com ciclodextrina. Os experimentos de fotoestabilidade indicam que a inserção da molécula de NF na cavidade interna da ciclodextrina protege o fármaco da fotodecomposição. A cinética de liberação mostra que o perfil de liberação do fármaco é modificado pela presença da ciclodextrina no meio. A estequiometria de complexação entre NF e HP-Beta-CD determinada foi de 1:1 NF:HP-Beta-CD. Os resultados demicroscopia eletrônica de varredura indicam alterações na estrutura cristalina da NF em presença deciclodextrina. Este estudo está baseado na caracterização físico-química da complexação entre NF e HP-Beta-CD podendo ser uma nova potencial opção para utilização terapêutica do NF.

Nitrofurazone (NF), 5-nitro-2-furaldehyde semicarbazone, a broad-spectrum antibiotic, has reported toxic effects and low solubility in water. It would be of great interest to form inclusion complexes between NF and a cyclodextrin, to develop more effective and safer antibiotic formulations. This paper focuses on the preparation of inclusion complexes of NF with 2- hydroxypropyl- -cyclodextrin (HP- -CD) and their initial characterization by evaluating rates of complex formation, photostability, solubility isotherms, release rate profiles, stoichiometry of the complexes and their morphology, as revealed by scanning electron microscopy. The kinetic tests of complex formation revealed that 17,3 h is enough for stabilization of the NFcyclodextrin complex. The solubility isotherm studies showed that the isotherm changes from type A to type B, as a function of temperature. The photostability experiments showed that the insertion of the NF in the HP- -CD cavity protects the drug from photodecomposition. The release kinetic tests showed that the profile of NF release from the complex is altered by the presence of HP- -CD in the medium. A Job's plot indicated that the stoichiometry of the complex was 1:1 NF:HP- -CD. The scanning electron micrographs showed changes in the crystal structure of NF in the complex. This study focused on the physicochemical properties of drug-delivery formulations that could potentially be developed into a novel type of therapy with NF.