Cellular consequences of small supernumerary marker chromosome derived from chromosome 12: mosaicism in daughter and father

Constitutional genomic imbalances are known to cause malformations, disabilities, neurodevelopmental delay, and dysmorphia and can lead to dysfunctions in the cell cycle. In extremely rare genetic conditions such as small supernumerary marker chromosomes (sSMC), it is important to understand the cellular consequences of this extra marker, as well the factors that contribute to their maintenance or elimination through successive cell cycles and phenotypic impact. The study of chromosomal mosaicism provides a natural model to characterize the effect of aneuploidy on genome stability and compare cells with the same genetic background and environment exposure, but differing in the presence of sSMC. Here, we report the functional characterization of different cell lines from two familial patients with mosaic sSMC derived from chromosome 12. We performed studies of proliferation dynamics, stability, and variability of these cells using fluorescent in situ hybridization (FISH), sister chromatid exchanges (SCE), and conventional staining. We also quantified the telomere-related genomic instability of sSMC cells using 3D telomeric profile analysis by quantitative-FISH. sSMC cells exhibited differences in the cell cycle dynamics compared to normal cells. First, the sSMC cells exhibited lower proliferation index and higher frequency of SCE than normal cells, associated with a higher level of chromosomal instability. Second, sSMC cells exhibited more telomeric-related genomic instability. Lastly, the differences of sSMC cells distribution among tissues could explain different phenotypic repercussions observed in patients. These results will help in our understanding of the sSMC stability, maintenance during cell cycle, and the cell cycle variables involved in the different phenotypic manifestations.

Multiresistance and virulence factors of Staphylococcus aureus isolated from pigs / [Multirresistência e fatores de virulência de Staphylococcus aureus isolados de suínos]

The emergence of livestock-associated methicillin-resistant Staphylococcus aureus strains (LA-MRSA) and the potential role of pigs in the evolution of these strains has led to increased interest in research of these microorganisms. However, this has contributed to a lack of research in the isolation and characterization of methicillin-susceptible S. aureus strains (MSSA). In this study, the prevalence of S. aureus in pigs in the nursery and finishing stages were analyzed. The susceptibility profiles to antibiotics, tolerance to heavy metals, and biofilm production of the isolates were evaluated using phenotypic and genotypic techniques. A total of 1,250 colonies suggestive of Staphylococcus spp. were isolated from 128 pigs, of which 63.6% (n = 795) belonged to this microbial genus. Sixty-seven colonies isolated from 34 animals (26.5%) were confirmed as S. aureus (8.4%). No strains resistant to copper, zinc, or methicillin were detected; however, all strains presented a resistance profile to at least three different classes of antimicrobials and 21 produced biofilms. These data are of concern, as they indicate the need for increased surveillance in the use of antimicrobials as well as reinforce the importance of studies on MSSA strains.(AU)

A emergência de cepas de Staphylococcus aureus resistentes à meticilina associadas à pecuária (LA-MRSA) e o papel potencial dos suínos na evolução dessas cepas têm levado ao aumento do interesse na pesquisa desses microrganismos. No entanto, isso tem contribuído para a falta de estudos sobre o isolamento e a caracterização de cepas de S. aureus sensíveis à meticilina (MSSA). Neste estudo, foi analisada a prevalência de S. aureus em suínos nas fases de creche e terminação. Os perfis de suscetibilidade aos antibióticos, a tolerância a metais pesados e a produção de biofilme dos isolados foram avaliados por meio de técnicas fenotípicas e genotípicas. Um total de 1.250 colônias sugestivas de Staphylococcus spp. foi isolado de 128 suínos, das quais 63,6% (n = 795) pertenciam a esse gênero microbiano. Sessenta e sete colônias isoladas de 34 animais (26,5%) foram confirmadas como S. aureus (8,4%). Nenhuma cepa resistente ao cobre, ao zinco ou à meticilina foi detectada; entretanto, todas as cepas apresentaram perfil de resistência a pelo menos três classes diferentes de antimicrobianos e 21 produziam biofilme. Esses dados são preocupantes, pois indicam a necessidade de maior vigilância no uso de antimicrobianos, bem como reforçam a importância de estudos com cepas de MSSA.(AU)

Antioxidant effect of the pequi oil (Caryocar brasiliense) on the hepatic tissue of rats trained by exhaustive swimming exercises / Efeito antioxidante do óleo de pequi (Caryocar brasiliense ) no tecido hepático de ratos treinados por exercícios de natação exaustivos

Abstract Increased oxygen consumption and activation of specific metabolic pathways during or after physical exercise lead to the formation of reactive oxygen and nitrogen species. An investigation was made into the effects of pequi oil supplementation in protecting liver cells against injury resulting from oxidative stress. The experiments involved 20 male adult Wistar rats ( Rattus norvegicus). The animals were divided into four experimental groups: Group 1: sedentary control group; Group 2: exercise control group; Group 3: supplemented sedentary group; and Group 4: supplemented exercise group. Supplementation consisted of pequi oil administered by oral gavage (400 mg). The animals of the exercised groups were subjected to 20 swimming sessions for 5 weeks (with progressive increase of 10 minutes until exhaustion). Samples were collected from the right hepatic lobe for histopathological analysis and determination of malondialdehyde levels. The histopathological analyses revealed that the animals of the exercised control group had moderate liver damage, while the animals of the supplemented exercised group had slight tissue damage, and the sedentary control and sedentary supplemented groups showed no tissue damage. The malondialdehyde levels showed higher and statistically significant in exercise control group when compared to the other evaluated groups (p<0.05). In conclusion the supplementation with pequi oil had a protective effect on liver cells against damage caused by oxygen free radicals during strenuous exercise, as demonstrated by the indicator of lipid peroxidation.

Resumo Aumento do consumo de oxigênio e ativação de vias metabólicas específicas durante ou após a atividade física conduz para formação de espécies reativas de oxigênio e nitrogênio. Uma investigação foi realizada sobre os efeitos da suplementação com óleo de pequi na proteção das células hepáticas contra lesões resultantes do estresse oxidativo. Na realização dos experimentos foram utilizados 20 ratos machos adultos da linhagem Wistar (Rattus novergicus ). Os animais foram divididos em quatro grupos experimentais: grupo 1: grupo sedentário controle; grupo 2: grupo treinado controle; grupo 3: grupo sedentário suplementado e grupo 4: grupo treinado suplementado. Na suplementação foi utilizado o óleo de pequi ministrado por gavagem oral (400 mg). Os animais dos grupos treinados foram submetidos a 20 sessões de natação por um período de 5 semanas (com aumento progressivo de 10 minutos até a exaustão). Foram retiradas amostras do lobo hepático direito para análises histopatológicas, e dosagem de malondialdeído. As análises histopatológicas revelaram que os animais do grupo treinado controle tiveram danos hepáticos moderados; já os animais do grupo treinado suplementado tiveram danos teciduais leves; os grupos sedentário controle e sedentário suplementado não apresentaram injúrias teciduais. Os níveis de malondialdeído mostraram-se maiores e estatisticamente significativos no grupo treinado controle quando comparados aos outros grupos avaliados (p<0,05). Podemos concluir que a suplementação com óleo de pequi teve efeito protetor nas células hepáticas contra os danos causados pelos radicais livres de oxigênio durante os exercícios exaustivos, conforme demonstrado pelo indicador de peroxidação lipídica.

Using in situ flow cytometry images of ciliates and dinoflagellates for aquatic system monitoring / Usando imagem de citometria de fluxo in situ de ciliados e dinoflagelados para o monitoramento de sistemas aquáticos

Abstract Short-period variability in plankton communities is poorly documented, especially for variations occurring in specific groups in the assemblage because traditional analysis is laborious and time-consuming. Moreover, it does not allow the high sampling frequency required for decision making. To overcome this limitation, we tested the submersible CytoSub flow cytometer. This device was anchored at a distance of approximately 10 metres from the low tide line at a depth of 1.5 metres for 12 hours to monitor the plankton at a site in the biological reserve of Barra da Tijuca beach, Rio de Janeiro. Data analysis was performed with two-dimensional scatter plots, individual pulse shapes and micro images acquisition. High-frequency monitoring results of two interesting groups are shown. The abundance and carbon biomass of ciliates were relatively stable, whereas those from dinoflagellates were highly variable along the day. The linear regression of biovolume measures between classical microscopy and in situ flow cytometry demonstrate high degree of adjustment. Despite the success of the trial and the promising results obtained, the large volume of images generated by the method also creates a need to develop pattern recognition models for automatic classification of in situ cytometric images.

Resumo A variabilidade de curto período em comunidades do plâncton é pouco documentada, especialmente as variações que ocorrem em grupos específicos das assembleias por causa das análises tradicionais serem muito trabalhosas e demoradas. Além disso, não permitem que a alta frequência amostral necessária para a tomada de decisão. Para superar esta limitação, nós testamos o CytoSub, um citômetro de fluxo submersível. Este aparelho foi ancorado a aproximadamente 10 metros de distância da linha de maré baixa a uma profundidade de 1,5 metros por 12 horas para monitorar o plâncton em um sítio da reserva biológica da praia da Barra da Tijuca, Rio de Janeiro. A análise dos dados foi realizada a partir de gráficos de dispersão bidimensionais, pelas assinaturas ópticas individuais escaneadas (pulse shape profile) e aquisição de micro imagens. Resultados do monitoramento de alta frequência de dois grupos interessantes são apresentados. A abundância e a biomassa de carbono de um grupo de ciliados foram relativamente estáveis, ao passo que o grupo de dinoflagelado, foi altamente variável ao longo do dia. O modelo de regressão linear das medidas de biovolume entre a clássica microscopia e a citometria de fluxo in situ apresentou alto grau de ajustamento. Apesar do sucesso deste ensaio e dos resultados promissores obtidos, o grande volume de imagens geradas por este método também gerou a necessidade de se desenvolver modelos de reconhecimento de padrões para a classificação automática de imagens de citometria in situ.

Using in situ flow cytometry images of ciliates and dinoflagellates for aquatic system monitoring

Abstract Short-period variability in plankton communities is poorly documented, especially for variations occurring in specific groups in the assemblage because traditional analysis is laborious and time-consuming. Moreover, it does not allow the high sampling frequency required for decision making. To overcome this limitation, we tested the submersible CytoSub flow cytometer. This device was anchored at a distance of approximately 10 metres from the low tide line at a depth of 1.5 metres for 12 hours to monitor the plankton at a site in the biological reserve of Barra da Tijuca beach, Rio de Janeiro. Data analysis was performed with two-dimensional scatter plots, individual pulse shapes and micro images acquisition. High-frequency monitoring results of two interesting groups are shown. The abundance and carbon biomass of ciliates were relatively stable, whereas those from dinoflagellates were highly variable along the day. The linear regression of biovolume measures between classical microscopy and in situ flow cytometry demonstrate high degree of adjustment. Despite the success of the trial and the promising results obtained, the large volume of images generated by the method also creates a need to develop pattern recognition models for automatic classification of in situ cytometric images.

Resumo A variabilidade de curto período em comunidades do plâncton é pouco documentada, especialmente as variações que ocorrem em grupos específicos das assembleias por causa das análises tradicionais serem muito trabalhosas e demoradas. Além disso, não permitem que a alta frequência amostral necessária para a tomada de decisão. Para superar esta limitação, nós testamos o CytoSub, um citômetro de fluxo submersível. Este aparelho foi ancorado a aproximadamente 10 metros de distância da linha de maré baixa a uma profundidade de 1,5 metros por 12 horas para monitorar o plâncton em um sítio da reserva biológica da praia da Barra da Tijuca, Rio de Janeiro. A análise dos dados foi realizada a partir de gráficos de dispersão bidimensionais, pelas assinaturas ópticas individuais escaneadas (pulse shape profile) e aquisição de micro imagens. Resultados do monitoramento de alta frequência de dois grupos interessantes são apresentados. A abundância e a biomassa de carbono de um grupo de ciliados foram relativamente estáveis, ao passo que o grupo de dinoflagelado, foi altamente variável ao longo do dia. O modelo de regressão linear das medidas de biovolume entre a clássica microscopia e a citometria de fluxo in situ apresentou alto grau de ajustamento. Apesar do sucesso deste ensaio e dos resultados promissores obtidos, o grande volume de imagens geradas por este método também gerou a necessidade de se desenvolver modelos de reconhecimento de padrões para a classificação automática de imagens de citometria in situ.

Antioxidant effect of the pequi oil (Caryocar brasiliense) on the hepatic tissue of rats trained by exhaustive swimming exercises

Abstract Increased oxygen consumption and activation of specific metabolic pathways during or after physical exercise lead to the formation of reactive oxygen and nitrogen species. An investigation was made into the effects of pequi oil supplementation in protecting liver cells against injury resulting from oxidative stress. The experiments involved 20 male adult Wistar rats ( Rattus norvegicus). The animals were divided into four experimental groups: Group 1: sedentary control group; Group 2: exercise control group; Group 3: supplemented sedentary group; and Group 4: supplemented exercise group. Supplementation consisted of pequi oil administered by oral gavage (400 mg). The animals of the exercised groups were subjected to 20 swimming sessions for 5 weeks (with progressive increase of 10 minutes until exhaustion). Samples were collected from the right hepatic lobe for histopathological analysis and determination of malondialdehyde levels. The histopathological analyses revealed that the animals of the exercised control group had moderate liver damage, while the animals of the supplemented exercised group had slight tissue damage, and the sedentary control and sedentary supplemented groups showed no tissue damage. The malondialdehyde levels showed higher and statistically significant in exercise control group when compared to the other evaluated groups (p 0.05). In conclusion the supplementation with pequi oil had a protective effect on liver cells against damage caused by oxygen free radicals during strenuous exercise, as demonstrated by the indicator of lipid peroxidation.

Resumo Aumento do consumo de oxigênio e ativação de vias metabólicas específicas durante ou após a atividade física conduz para formação de espécies reativas de oxigênio e nitrogênio. Uma investigação foi realizada sobre os efeitos da suplementação com óleo de pequi na proteção das células hepáticas contra lesões resultantes do estresse oxidativo. Na realização dos experimentos foram utilizados 20 ratos machos adultos da linhagem Wistar (Rattus novergicus ). Os animais foram divididos em quatro grupos experimentais: grupo 1: grupo sedentário controle; grupo 2: grupo treinado controle; grupo 3: grupo sedentário suplementado e grupo 4: grupo treinado suplementado. Na suplementação foi utilizado o óleo de pequi ministrado por gavagem oral (400 mg). Os animais dos grupos treinados foram submetidos a 20 sessões de natação por um período de 5 semanas (com aumento progressivo de 10 minutos até a exaustão). Foram retiradas amostras do lobo hepático direito para análises histopatológicas, e dosagem de malondialdeído. As análises histopatológicas revelaram que os animais do grupo treinado controle tiveram danos hepáticos moderados; já os animais do grupo treinado suplementado tiveram danos teciduais leves; os grupos sedentário controle e sedentário suplementado não apresentaram injúrias teciduais. Os níveis de malondialdeído mostraram-se maiores e estatisticamente significativos no grupo treinado controle quando comparados aos outros grupos avaliados (p 0,05). Podemos concluir que a suplementação com óleo de pequi teve efeito protetor nas células hepáticas contra os danos causados pelos radicais livres de oxigênio durante os exercícios exaustivos, conforme demonstrado pelo indicador de peroxidação lipídica.

Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus

Biofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaining intact RNA preparations is the first and most critical step for these studies. In this article, we describe an optimized protocol for obtaining total RNA from sessile cells of S. aureus using the RNeasy Mini Kit. This method essentially consists of a few steps, as follows: 1) addition of acetone-ethanol to sessile cells, 2) lysis with lysostaphin at 37°C/10 min, 3) vigorous mixing, 4) three cycles of freezing and thawing, and 5) purification of the lysate in the RNeasy column. This simple pre-kit procedure yields high-quality total RNA from planktonic and sessile cells of S. aureus.

Qualidade da carne de bovinos superprecoces terminados em confinamento / Quality of meat from super-young cattle finished in feedlot

Avaliou-se a qualidade da carne de 10 bovinos F1 Guzerá-Nelore e 10 F2 Pardo Suíço-Guzerá-Nelore, inteiros, e abatidos precocemente. Na avaliação da carne, foi utilizado o músculo Longissimus dorsi. O delineamento experimental foi o inteiramente ao acaso. Verificaram-se para F1 e F2, respectivamente, médias para umidade de 75,3 e 75,1, extrato etéreo de 1,4 e 1,5, proteína de 23,7 e 23,6 e cinzas de 1,04 e 1,06. Cor em L* de 28,9 e 30,3, médias para cor em a* de 22,6 e 23,4 e cor em b* de 15,3 15,4, força de cisalhamento de 3,4 e 3,5, perda de peso pelo descongelamento de 9,8 e 10,5 e perda pelo cozimento de 15,9 e 15,6. Não houve diferença estatística para nenhuma das variáveis analisadas entre os grupos genéticos.

The purpose of this study was to use instrumental analysis to evaluate the chemical composition and fatty acid profile of 10 cattle Guzerath-Nellore (F1) and 10 Brown Swiss-Guzerath-Nellore (F2), intact and feedlot finished and slaughtered at 13 months. The Longissimus dorsi was used for meat evaluation. The experimental design was completely randomized. Humidity for F1 and F2 was respectively 75,3 and 75,1 averages, 1,4 and 1,5 averages for ether extract, proteins were 23,7 and 23,6 and 1,04 and 1,06 averages for ash. The results for F1 and F2, respectively, with averages for color to L * of 28,9 and 30,3, averages for the color to a* of 22,6 and 23,4 and color to b * 15,3 and 15,4, averages for shear force of 3,4 and 3,4, weight loss by thawing of 9,8 and 10,5 and weight lost by cooking of 15,9 and 15,4. There were no differences in genetic groups analysis.

Emergence of clonal complex 5 (CC5) methicillin-resistant Staphylococcus aureus (MRSA) isolates susceptible to trimethoprim-sulfamethoxazole in a Brazilian hospital

In this study, genotyping techniques including staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and restriction-modification tests were used to compare the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered at two times within a 10-year interval (1998 and 2008) from a tertiary Brazilian hospital. In addition, the antimicrobial susceptibility profiles were analyzed. All 48 MRSA isolates from 1998 and 85.7% from 2008 (48/56 isolates) displayed multidrug-resistance phenotypes and SCCmec III. All but one of the 13 representative SCCmec III isolates belonged to CC8 and had PFGE patterns similar to that of the BMB9393 strain (Brazilian epidemic clone of MRSA; BEC). In 2008, we found an increased susceptibility to rifampicin and chloramphenicol among the SCCmec III isolates. In addition, we detected the entrance of diverse international MRSA lineages susceptible to trimethoprim-sulfamethoxazole (SXT), almost all belonging to CC5. These non-SCCmec III isolates were related to the USA 300 (ST8-SCCmec IV; PFGE-type B), USA 800 (ST5-SCCmec IV; subtype D1), USA 100 (ST5-SCCmec II; subtype D2), and EMRSA-3/Cordobes (ST5-SCCmec I, type C) clones. To the best of our knowledge, this is the first report of the emergence of isolates genetically related to the EMRSA-3/Cordobes clone in southeast Brazil. In this regard, these isolates were the most common non-SCCmec III MRSA in our institution, accounting for 8.9% of all isolates recovered in 2008. Thus, despite the supremacy of BEC isolates in our country, significant changes may occur in local MRSA epidemiology, with possible consequences for the rationality of MRSA empiric therapy.

Antimicrobial activity, cytotoxicity and intracellular growth inhibition of Portuguese Thymus essential oils

Thyme essential oils are well recognized by their excellent biological activities and the antimicrobial activity of Portuguese thyme essential oils has been investigated with promising results, particularly against food borne pathogens. In this study the potential antimicrobial activity of the essential oils of five species of Thymus (Lamiaceae), namely Th. caespititius Brot., Th. camphoratus Hoffmanns. & Link, Th. capitellatus Hoffmanns. & Link., Th. carnosus Boiss. and Th. zygis L. was evaluated against Candida albicans, Haemophilus influenza, Helicobacter pylori, Listeria monocytogenes, Salmonella enterica and Streptococcus pneumoniae. H. pylori strains were the most susceptible bacteria, particularly to the essential oils of Th. caespititius (Planalto Central), Th. zygis (Rebordãos) and Th. caespititius (Pico) which minimum inhibitory concentration (MIC) values ranged from 0.05 to 0.08 mg.mL-1. Th. caespititius essential oil from Planalto Central or its main component, carvacrol significantly (p<0.05) inhibited the intracellular growth of H. pylori, and showed no citotoxicity to the gastric cell line. Our results suggest the potential of this essential oil and its main component as a promising tool as anti-Helicobacter agent potentiating the eradication of this important gastroduodenal pathogen.

Virioplankton abundance in trophic gradients of an upwelling field

This work correlates time series of biological and physical variables to the marine viruses across trophic gradients within Arraial do Cabo upwelling system, Southeast of Brazil. The objective is to investigate the major controlling factors of virioplankton dynamics among different water masses. It was used an in situ and ex situ flow cytometry for accessing the plankton community. Viruses were highly correlated to bacteria and phytoplankton, but although the lack of direct correlation with physicals, upwelling turned out to be the main contributing factor to the highest values of viral abundance and virus:bacterial ratio. Our data suggest that the lowest temperature of upwelled South Atlantic Central Waters would help to maintain a high viral abundance and higher temperatures of Coastal and Tropical Waters might be another ecological niche allowing the co-existence.

Genotyping of methicillin-resistant Staphylococcus aureus isolates obtained in the Northeast region of Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) is a major agent of hospital infections worldwide. In Brazil, a multiresistant MRSA lineage (ST239-SCCmecIIIA), the so-called Brazilian epidemic clone (BEC), has predominated in all regions. However, an increase in nosocomial infections caused by non-multiresistant MRSA clones has recently been observed. In the present study, 45 clinical isolates of MRSA obtained from a university hospital located in Natal city, Brazil, were identified by standard laboratory methods and molecularly characterized using staphylococcal chromosome cassette mec (SCCmec) typing and pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was carried out using CLSI methods. The MRSA isolates studied displayed a total of 8 different pulsed-field gel electrophoresis patterns (types A to H) with predominance (73 percent) of pattern A (BEC-related). However, MRSA harboring SCCmec type IV were also identified, 3 (7 percent) of which were genetically related to the pediatric clone - USA800 (ST5-SCCmecIV). In addition, we found a considerable genetic diversity within BEC isolates. MRSA displaying SCCmecIV are frequently susceptible to the majority of non-β-lactam antibiotics. However, emergence of multiresistant variants of USA800 was detected.

The first report in Brazil of severe infection caused by community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA)

Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emergent pathogen in Brazil. However, there are no data on the prevalence of CA-MRSA. We report here the first well-characterized case of severe life-threatening CA-MRSA infection in a child living in Rio de Janeiro city. The patient had many complications including hematogenous osteomyelitis and involvement of multiple sites requiring drainage of soft-tissue abscess, and pleural and pericardial empyema. The MRSA isolates recovered were genotyped using PFGE, SCCmec typing and multilocus sequence typing. Disk diffusion tests were performed following Clinical and Laboratory Standards Institute recommendations. In addition, the presence of Panton-Valentine leukocidin (PVL) was assessed by PCR amplification, using specific primers for lukF-pv (encoding for the F subunit of the PVL). The bacterial isolates were related to the ST30-SCCmecIV lineage (Oceania Southwest Pacific clone), a PVL producer CA-MRSA previously detected in Porto Alegre, RS, Brazil. Also, the isolates analyzed were susceptible to all non-â-lactam antibiotics tested. The present report demonstrates that disseminated CA-MRSA disease is also occurring in Rio de Janeiro. Thus, the empirical treatment of moderate or severe infections suspected of being associated with CA-MRSA needs to be reviewed in order to allow prompt initiation of an effective therapy that also covers these microorganisms.

PtSRR1, a putative Pisolithus tinctorius symbiosis related receptor gene is expressed during the first hours of mycorrhizal interaction with Castanea sativa roots / PtSRR1, um possível receptor simbiose-regulado de Pisolithus tinctorius é expresso nas primeiras horas de interação ectomicorrízica com raízes de Castanea sativa

PtSRR1 EST was previously identified in the first hours of Pisolithus tinctorius and Castanea sativa interaction. QRT-PCR confirmed PtSRR1 early expression and in silico preliminary translated peptide analysis indicated a strong probability that PtSRR1 be a transmembrane protein. These data stimulate the PtSRR1 gene research during ectomycorrhiza formation.

PtSRR1 foi isolado preliminarmente de P. tinctorius nas primeiras horas da interação com raízes de C. sativa. Análises de QRT-PCR confirmaram sua expressão positiva (12 h) e seu peptídeo putativo indicou forte possibilidade para proteína transmembranar. Estes dados estimulam o estudo do PtSRR1 durante a formação de ectomicorrizas.

Análise da atividade antimicrobiana das folhas de Davilla elliptica St.-Hil. (Dilleniaceae) / Investigation of antimicrobial activity of Davilla elliptica St.-Hil. leaves (Dilleniaceae)

Davilla elliptica St.-Hil. (Dilleniaceae) é um arbusto,usado popularmente como adstringente, tônico, sedativo, diurético, no tratamento de hemorróidas, hérnia e em aplicações tópicas como anti-séptico na limpeza de ferimentos. O presente estudo teve por objetivo avaliara atividade antimicrobiana dos extratos etanólicos brutos das folhas de D. elliptica coletadas em Alexânia e Goiânia, Goiás, Brasil. Os extratos etanólicos brutos foram obtidos, por maceração a frio, a partir das folhas dessecadas e pulverizadas. A atividade antimicrobiana foi avaliada utilizando 25 cepas de microrganismos (Gram-negativos, Gram-positivos e levedura) através da determinação da Concentração Inibitória Mínima (CIM) utilizando-se o inoculador de Steers. Os extratos obtidos de plantas coletados nas duas localidades inibiram o crescimento de todas as cepas avaliadas. O extrato da amostra coletada em Alexânia apresentou atividade antimicrobiana com CIM variando de 0,371 mg/mL a 0,743 mg/mL. O extrato da amostra coletada em Goiânia apresentou CIM variando de < 0,023 mg/mL a 11,9 mg/mL. Os extratos etanólicos brutos das folhas de D. elliptica foram mais ativos contra as bactérias Gram-positivas do que para as bactérias Gram-negativas.

Davilla elliptica St.-Hil.(Dilleniaceae) is a shrub, whose leaves are traditionally used in Brazilian popular medicine as an astringent, tonic, sedative and diuretic and to treat hemorrhoids and hernia and it is applied a topical antiseptic to clean wounds. The aim in this study was to assess the antimicrobial activity of crude ethanol extracts of D. elliptica leaves collected in the towns of Alexânia and Goiânia, state of Goiás, Brazil. The crude extracts were obtained by cold maceration, of the dried and pulverized leaves in ethanol. Antimicrobial activities against 25 strains of microorganisms (Gram-negative bacteria, Gram-positive bacteria and a yeast) were tested determining by the Minimal Inhibitory Concentration (MIC) in agar plates inoculated with using a Steers replicator. Extracts of from inhibited the growth of all strains evaluated. The extract of the sample collected in Alexânia showed antimicrobial activity with MICs ranging from 0.371 mg/mL to 0.743 mg/mL, while that collected in Goiânia showed MICs ranging from <0.023 mg/mL to 11.9 mg/mL. The D. elliptica crude leaf ethanol extracts evaluated were more active against Gram-positive than Gram-negative bacteria.

Avaliação da atividade antimicrobiana das partes aéreas (folhas e caules) e raízes de Richardia brasiliensis Gomez (Rubiaceae) / Assessment of antimicrobial activity of aerial parts (leaves and stems) and roots of Richardia brasiliensis Gomez (Rubiaceae)

Richardia brasiliensis é uma planta utilizada popularmente como expectorante, emética, diaforética, vermífuga e para o tratamento de hemorróidas. O presente estudo objetivou realizar a prospecção fitoquímica e avaliar a ação antimicrobiana do extrato bruto das partes aéreas e raízes de R. brasiliensis. O extrato etanólico bruto foi obtido a partir do material botânico dessecado e pulverizado. O pó das partes aéreas e raízes foram submetidos à triagem fitoquímica. A atividade antimicrobiana foi realizada contra bactérias Gram-positivas esporuladas e não esporuladas, Gram negativas e a levedura Candida albicans através da determinação da concentração inibitória mínima por diluição em ágar empregando-se o inoculador de Steers. A triagem fitoquímica evidenciou a presença de esteróides, triterpenóides, cumarinas, resinas, alcaloides e flavonóides. A CIM do extrato das partes aéreas variou de 0,37 a 0,74 mg/mL e das raízes de 0,74 a 11,9 mg/mL. Concluiu-se que tanto o extrato etanólico bruto das partes aéreas quanto das raízes apresentaram atividade antimicrobiana.

Richardia brasiliensis is a neotropical perennial her bused in Brazilian popular medicine as an expectorant, emetic, diaphoretic, vermifuge and to treat hemorrhoids. This paper reports the phytochemical screening and the antimicrobial activity of crude ethanol extracts of the aerial parts and roots of this plant. The crude ethanol extracts were prepared from dried, pulverized botanical material. The aerial parts and roots powder was screened for classes of phytochemicals. The antimicrobial activity was tested against spore-forming and non-sporing Gram-positive bacteria, Gramnegative bacteria and Candida albicans by the agardilution method, inoculating the series of plates at multiple points with a Steers replicator to determine the minimum inhibitory concentration (MIC) of each organism. The phytochemical screening showed the presence of steroids, triterpenes, coumarins, resins, alkaloids and flavonoids. The MIC of the extract of aerial parts varied from 0.37 to 0.74 mg/mL and that of the roots from 0.74 to 11.9 mg/mL. It was concluded that the crude ethanolic extracts of both the aerial parts and the roots showed antimicrobial activity.

Breathing disorders in congestive heart failure: gender, etiology and mortality

We investigated the relationship between sleep-disordered breathing (SDB) and Cheyne-Stokes respiration (CSR) while awake as well as mortality. Eighty-nine consecutive outpatients (29 females) with congestive heart failure (CHF; left ventricular ejection fraction, LVEF <45 percent) were prospectively evaluated. The presence of SDB and of CSR while awake before sleep onset was investigated by polysomnography. SDB prevalence was 81 and 56 percent, using apnea-hypopnea index cutoffs >5 and >15, respectively. CHF etiologies were similar according to the prevalence of SDB and sleep pattern. Males and females were similar in age, body mass index, and LVEF. Males presented more SDB (P = 0.01), higher apnea-hypopnea index (P = 0.04), more light sleep (stages 1 and 2; P < 0.05), and less deep sleep (P < 0.001) than females. During follow-up (25 ± 10 months), 27 percent of the population died. Non-survivors had lower LVEF (P = 0.01), worse New York Heart Association (NYHA) functional classification (P = 0.03), and higher CSR while awake (P < 0.001) than survivors. As determined by Cox proportional model, NYHA class IV (RR = 3.95, 95 percentCI = 1.37-11.38, P = 0.011) and CSR while awake with a marginal significance (RR = 2.96, 95 percentCI = 0.94-9.33, P = 0.064) were associated with mortality. In conclusion, the prevalence of SDB and sleep pattern of patients with Chagas' disease were similar to that of patients with CHF due to other etiologies. Males presented more frequent and more severe SDB and worse sleep quality than females. The presence of CSR while awake, but not during sleep, may be associated with a poor prognosis in patients with CHF.

Caracterização farmacognóstica de folhas de Davilla elliptica St.-Hil. (Dilleniaceae) / Pharmacognostic characterization of the leaves of Davilla elliptica St.-Hil. (Dilleniaceae)

Davilla elliptica St.-Hil. é empregada popularmente no tratamento de hemorróidas, diarréias e ferimentos. O propósito deste trabalho foi realizar o estudo farmacognóstico de folhas de D. elliptica de dois locais do Estado de Goiás. Cortes paradérmicos e transversais foram submetidos a técnicas usuais de microscopia óptica. As folhas de D. elliptica são hipoestomáticas, sendo os estômatos predominantemente paracíticos. Na epiderme observam-se tricomas unicelulares em ambas faces. O mesofilo é isobilateral e apresenta idioblastos cristalíferos. O sistema vascular na nervura principal é constituído por três feixes vasculares, formando uma estrutura circular. No pecíolo o sistema vascular apresenta uma estrutura circular formada, provavelmente, pela união de vários feixes vasculares. O material pulverizado foi submetido a testes de prospecção fitoquímica, de pureza e quantitativo. Foi constatada a presença de taninos, cumarinas, resinas, flavonóides, saponinas, esteróides e triterpenóides. Os teores de cinzas totais foram de 10,32 por cento e 12,26 por cento; os de cinzas insolúveis em ácido, 7,46 por cento e 9,68 por cento; e os de umidade, 9,92 por cento e 9,50 por cento. Os teores de flavonóides totais foram de 1,20 por cento e 1,48 por cento, e os de taninos 9,89 por cento e 13,96 por cento. Devido ao número reduzido de amostras, não é possível afirmar, através deste estudo, que o perfil farmacognóstico de D. elliptica seja influenciado por fatores abióticos.

Davilla elliptica St.-Hil. is popularly used in the treatment of hemorrhoid, diarrhea and wounds. The aim of this work was to characterize pharmacognosticaly two leaves samples of D. elliptica. Paradermal and transversal sections had been submitted the usual techniques of optical microscopy. The leaves of D. elliptica are hypostomatics, and the stomata predominantly paracytic. In both epidermis were observed unicellular non-glandular trichomes. The mesophyll is isolateral and shows idioblasts with crystals in raphide. The vascular system in the midrib is constituted by three vascular bundles, forming a circular structure. The petiole shows a circular structure probably formed by union of many vascular bundles localized in the central region. The powdered plant material was submitted to tests of phytochemical prospection, of purity and quantitative. There had been evidenced the presence of tannins, coumarins, resins, flavonoids, saponins and steroids. The content of total ashes was of 10,32 percent and 12,26 percent, insoluble in acid of 7,46 percent and 9,68 percent; the humidity of 9,92 percent and 9,50 percent. The flavonoids contents was of 1,20 percent and 1,48 percent; the tannins contents was of 9,89 percent and 13,96 percent. Because of the reduced number of samples, it was not possible to affirm through this study that the pharmacognostic profile de D. elliptica is influenced by abiotics factors.

Linear competitive inhibition of human tissue kallikrein by 4-aminobenzamidine and benzamidine and linear mixed inhibition by 4-nitroaniline and aniline

Hydrolysis of D-valyl-L-leucyl-L-arginine p-nitroanilide (7.5-90.0 `M) by human tissue kallikrein (hK1) (4.58-5.27 nM) at pH 9.0 and 37ºC was studied in the absence and in the presence of increasing concentrations of 4-aminobenzamidine (96-576 `M), benzamidine (1.27-7.62 mM), 4-nitroaniline (16.5-66 `M) and aniline (20-50 mM). The kinetic parameters determined in the absence of inhibitors were: Km = 12.0 + or - 0.8 `M and k cat = 48.4 + or - 1.0 min-1. The data indicate that the inhibition of hK1 by 4-aminobenzamidine and benzamidine is linear competitive, while the inhibition by 4-nitroaniline and aniline is linear mixed, with the inhibitor being able to bind both to the free enzyme with a dissociation constant Ki yielding an EI complex, and to the ES complex with a dissociation constant Ki', yielding an ESI complex. The calculated Ki values for 4-aminobenzamidine, benzamidine, 4-nitroaniline and aniline were 146 + or - 10, 1,098 + or - 91, 38.6 + or - 5.2 and 37,340 + or - 5,400 `M, respectively. The calculated Ki' values for 4-nitroaniline and aniline were 289.3 + or - 92.8 and 310,500 + or - 38,600 `M, respectively. The fact that Ki'>Ki indicates that 4-nitroaniline and aniline bind to a second binding site in the enzyme with lower affinity than they bind to the active site. The data about the inhibition of hK1 by 4-aminobenzamidine and benzamidine help to explain previous observations that esters, anilides or chloromethyl ketone derivatives of Nalpha-substituted arginine are more sensitive substrates or inhibitors of hK1 than the corresponding lysine compounds

Absorçäo real, perdas endógenas e exigência de fósforo para suínos em final de crescimento, pela técnica da diluiçäo isotópica / True absorption and endogenous fecal losses of phosphorus for swine in the final of the growing based on the isotopic dilution technique

Determinaram-se as perdas endógenas mínimas, a exigência de fósforo e a absorçäo real do fósforo do fosfato bicálcico em suínos no final da fase de crescimento, pela técnica da diluiçäo isotópica. Foram utilizados 20 leitöes híbridos comerciais, com peso de 48kg, distribuídos em delineamento experimental de blocos ao acaso com cinco tratamentos, constituídos de dietas com diferentes níveis de P total (0,33, 0,36, 0,47, 0,54 e 0,61 por cento), com quatro repetiçöes. A excreçäo fecal e urinária, a absorçäo e retençäo de P foram influenciadas pelos níveis de consumo desse mineral. As perdas endógenas fecais näo foram influenciadas pelos níveis de P consumido, indicando que os níveis ingeridos foram insuficientes para atender às exigências de P dos leitöes. A biodisponibilidade do fósforo bicálcico foi de 74,7 por cento e a exigência de mantença de 10,02mgP/kg p.v./dia