Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep

PURPOSE: To evaluate different protocols to isolate stem cells from ovine umbilical cord blood and adipose tissue. METHODS: There were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. There were used 3 protocols for obtainment and culture of umbilical cord blood stem cells and 4 protocols for ovine adipose tissue stem cells. RESULTS: It was possible to observe only one successful protocol for the obtainment of umbilical cord blood stem cells. When analyzing the techniques used to obtain adipose tissue stem cells, only one of the methods was effective as well. Through colony forming unit assay, there were obtained 58 colonies of cells after seven days in culture. Flow citometry tests revealed the cells were positive to CD44 and exhibited negative reaction to CD38, CD45, CD41/61. These cells showed a growth curve with very well defined phases LOG, LAG and PLATEAU. This phases are typically seem in mesenchymal stem cells growth curves. CONCLUSIONS: The isolation and culture of mesenchymal stem cells from ovine umbilical cord blood are complex and request more detailed assays. Stem cells from fat tissue sheep showed mesenchymal characteristics, according to their cell growth curve, ability to origin colonies of fibroblastoid cells and positive reactivity with the antibody CD44 by flow citometry.

Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p<0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and viceversa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol.

Scavenging processes of atmospheric particulate matter: a numerical modeling of case studies

Below cloud scavenging processes have been investigated considering a numerical simulation, local atmospheric conditions and particulate matter (PM) concentrations, at different sites in Germany. The below cloud scavenging model has been coupled with bulk particulate matter counter TSI (Trust Portacounter dataset, consisting of the variability prediction of the particulate air concentrations during chosen rain events. The TSI samples and meteorological parameters were obtained during three winter Campaigns: at Deuselbach, March 1994, consisting in three different events; Sylt, April 1994 and; Freiburg, March 1995. The results show a good agreement between modeled and observed air concentrations, emphasizing the quality of the conceptual model used in the below cloud scavenging numerical modeling. The results between modeled and observed data have also presented high square Pearson coefficient correlations over 0.7 and significant, except the Freiburg Campaign event. The differences between numerical simulations and observed dataset are explained by the wind direction changes and, perhaps, the absence of advection mass terms inside the modeling. These results validate previous works based on the same conceptual model.

Wives of pathological gamblers: personality traits, depressive symptoms and social adjustment

Objective: Wives of pathological gamblers tend to endure long marriages despite financial and emotional burden. Difficulties in social adjustment, personality psychopathology, and comorbidity with psychiatric disorders are pointed as reasons for remaining on such overwhelming relationships. The goal was to examine the social adjustment, personality and negative emotionality of wives of pathological gamblers. Method: The sample consisted of 25 wives of pathological gamblers, mean age 40.6, SD = 9.1 from a Gambling Outpatient Unit and at GAM-ANON, and 25 wives of non-gamblers, mean age 40.8, SD = 9.1, who answered advertisements placed at the Universidade de São Paulo hospital and medical school complex. They were selected in order to approximately match demographic characteristics of the wives of pathological gamblers. Subjects were assessed by the Social Adjustment Scale, Temperament and Character Inventory, Beck Depression Inventory and State-Trait Anxiety Inventory. Results: Three variables remained in the final Multiple Logistic Regression model, wives of pathological gamblers presented greater dissatisfaction with their marital bond, and higher scores on Reward Dependence and Persistence temperament factors. Both, Wives of pathological gamblers and wives of non-gamblers presented well-structured character factors excluding personality disorders. Conclusion: This personality profile may explain wives of pathological gamblers emotional resilience and their marriage longevity. Co-dependence and other labels previously used to describe them may work as a double edged sword, legitimating wives of pathological gamblers problems, while stigmatizing them as inapt and needy.

Ichthyofauna of Rio Jurubatuba, Santos, São Paulo: a high diversity refuge in impacted lands

Ichthyofaunistic surveys in the Atlantic Rainforest have been published in relatively few works, in spite of the major biological importance of this once vast biome which is rapidly vanishing due to disordered human population growth and natural resources overexploitation. The present study aimed to access the fish fauna of a relatively well preserved basin between the cities of Santos and Cubatão (SP), an area highly modified by human activities where recent ichthyofaunistic surveys are still missing. Collections were made during three field trips in Rio Jurubatuba, a medium sized costal river, and Riacho Sabão, one of its main tributaries. A total of 2773 specimens were sampled, representing 25 species from 14 families. Six species were primary marine using the upper reaches of Rio Jurubatuba. Twelve of the 19 freshwater species are endemic of the Atlantic Rainforest and four are present in regional lists of endangered species. Only five species occurred in both Rio Jurubatuba and Riacho Sabão. The most diverse family was Characidae, followed by Poeciliidae, Rivulidae and Heptapteridae. Phalloceros caudimaculatus was the most abundant species, followed by Poecilia vivipara and Geophagus brasiliensis. The study area is considered well preserved and due to its critical location, urges for conservation policies to protect its fish diversity.

Ichthyoplankton distribution and abundance in the northern Todos os Santos and Camamu Bays, Bahia State - Brazil

The occurrence, distribution and abundance of ichthyoplankton in Todos os Santos and Camamu Bays were analyzed based on four samplings (winter 2003, summer 2003, winter 2004 and summer 2005). Samples were obtained by surface horizontal hauls, using a 200-µm mesh conical-cylinder plankton net. The distribution and abundance of eggs indicate a remarkable seasonal and annual variation of spawning activity in the region, especially when the two summer campaigns are compared. In summer 2003 the highest quantitative values were recorded, especially for Camamu, where the maximum reached 106.56 eggs.m-3, with an overall average of 43.46 eggs.m-3 for the two areas. In summer 2005 values were relatively low, the overall average being 3.49 eggs.m-3. The larval taxonomic composition is characterized by the predominance of gobiids, with small variation from summer to winter. Considering all the campaigns and samplings undertaken in both areas, larvae of 11 families were identified: Engraulidae, Clupeidae, Mugilidae, Atherinopsidae, Hemiramphidae, Syngnathidae, Blenniidae, Carangidae, Gobiidae, Achiridae and Tetraodontidae.

Ichthyofauna in an estuary of the Mataripe area, Todos os Santos Bay, Bahia, Brazil

The community structure and dynamics as well as some biological parameters of selected species of the ichthyofauna of the Mataripe estuarine region affected by the Landulfo Alves Oil Refinery (RLAM) were analyzed. Twenty stations were sampled with a gillnet in five different periods: August and December 2003, March and July/August 2004 and January 2005. Thirty-five actinopterygian species and one elasmobranch species were recorded, Oligoplites saurus, Diapterus rhombeus, Lutjanus synagris and Scomberomorus brasiliensis among them, on all the campaigns. A total of 1368 specimens, weighing 36.10 kg, were caught. The ichthyofauna total biomass was greater, in weight, on the eastern side of the study region, especially at the stations close to the shoals/reefs and the rocky bottom. A similar pattern was also observed for the diversity values. In general, low evenness and diversity were observed in the area studied, possibly as a result of the fishing gear used. D. rhombeus juveniles dominated in all but one of the samplings (July 2004), in which latter Cyclichthys spinosus was dominant. Carangids and species associated with consolidated bottoms were observed, although in small numbers, throughout the study period. In spite of the limitations imposed by the gear used for sampling, the estuarine area influenced by the RLAM was seen to play a role as a growth area for the great majority of species, especially the mojarra (D. rhombeus), but it offers no fishing potential due to the prevalence of young and small individuals. Evidence of imminent spawning was recorded for Pomadasys corvinaeformis in August 2003, and recent spawning in March 2004 for Oligoplites saurus. Further, mature individuals occurred in insufficient numbers to permit population level evaluation.

Hydrodynamics and suspended sediment transport in the Camboriú estuary - Brazil: pre jetty conditions

Estuarine hydrodynamics is a key factor in the definition of the filtering capacity of an estuary and results from the interaction of the processes that control the inlet morphodynamics and those that are acting in the mixing of the water in the estuary. The hydrodynamics and suspended sediment transport in the Camboriú estuary were assessed by two field campaigns conducted in 1998 that covered both neap and spring tide conditions. The period measured represents the estuarine hydrodynamics and sediment transport prior to the construction of the jetty in 2003 and provides important background information for the Camboriú estuary. Each field campaign covered two complete tidal cycles with hourly measurements of currents, salinity, suspended sediment concentration and water level. Results show that the Camboriú estuary is partially mixed with the vertical structure varying as a function of the tidal range and tidal phase. The dynamic estuarine structure can be balanced between the stabilizing effects generated by the vertical density gradient, which produces buoyancy and stratification flows, and the turbulent effects generated by the vertical velocity gradient that generates vertical mixing. The main sediment source for the water column are the bottom sediments, periodically resuspended by the tidal currents. The advective salt and suspended sediment transport was different between neap and spring tides, being more complex at spring tide. The river discharge term was important under both tidal conditions. The tidal correlation term was also important, being dominant in the suspended sediment transport during the spring tide. The gravitational circulation and Stokes drift played a secondary role in the estuarine transport processes.

Hidrodinâmica e transporte de material particulado em suspensão sazonal em um estuário dominado por maré: Estuário de Caravelas (BA)

O objetivo do presente estudo é caracterizar a hidrodinâmica e o transporte de material particulado em suspensão (MPS) no estuário de Caravelas sob diferentes condições de maré e vazão fluvial. Foram realizadas quatro campanhas hidrográficas durante ciclos completos de maré, sendo em condições de maré de sizígia e quadratura, e durante os períodos seco e chuvoso. Dados de nível de água, velocidade e direção de correntes, salinidade, temperatura e turbidez foram obtidos em uma estação fixa próxima da desembocadura do estuário. A partir destes dados foram obtidos os transportes residuais de MPS e calculado os mecanismos de transporte. As maiores concentrações médias de material particulado em suspensão ocorreram em condições de sizígia. Em condições de maré de quadraturao transporte resultante de MPS foi com sentido estuário acima, porém de pequena intensidade. Em condições de sizígia o estuário é caracterizado por correntes mais intensas e assimétricas, com dominância de vazante. Durante a condição de sizígia o estuário pode atuar como importador ou exportador de MPS. O estuário do rio Caravelas foi classificado como bem misturado e fracamente estratificado. A hidrografia e o balanço sedimentar são principalmente modulados pela altura da marée o aporte de água doce é irrelevante.

A descriptive analysis of the seasonal variation of physical oceanographic characteristics in the northern region of the Todos os Santos Bay (Bahia, Brazil)

The results obtained in the August and December 2003, August 2004 and January 2005 oceanographic campaigns in the northern region of the Todos os Santos Bay (lat. 12º44.5'S; long. 038º35.00'W) between the Madre de Deus and Maré islands are analyzed. Instruments of continuous and discrete samplings were used to measure hydrographic properties currents and tides. The water mass of the northern region of the bay is forced by semidiurnal and mesotides of form number 0.08 and the lunar component M2 height was estimated at 91cm. The time series of the surface currents indicated movements in the N/S direction, forced by the tide with maximum magnitudes of 0.73 m.s-1 on the December 2003 campaign. However, in August 2004 the currents were dominated by the wind stress forcing, with a maximum speed of 1.85 m.s-1 and SE direction. Near the bottom, the influence of the tide is not as evident, with a decrease in intensity due to internal and bottom friction, with a maximum velocity of 0.17 m.s-1. The thermal and haline structures were weakly horizontally, as well as vertically stratified, with extreme values varying in the intervals 23ºC (August, 2004) to 28ºC (December, 2003) and 31.0 psu (August, 2003) to 36.0 psu (December, 2003), respectively. Some conclusions may be drawn from these results: i) The signs of the dilution of the fresh water discharges of the Caípe, Mataripe and São Paulo rivers in the region under the influence of the RLAM were observed only during the winter periods, but in the summer the region was flooded by waters of oceanic origin and the salinities above 36.0 indicated TW mass intrusion; ii) The N-S circulation near the RLAM is strongly dominated by the tide, and the importance of the M2 component was unequivocal, however, the E-W component presented some tidal modulation away from abrupt bottom topographical changes, and iii) The residual series, calculated as the difference between the original and modeled, is about ¼ of the original and confirmed its semidiurnal character.

Population structure of the lancelet Branchiostoma caribaeum (Cephalochordata: Branchiostomidae) in the Baía de Guanabara, Rio de Janeiro, southeastern Brazil

Population structure of the lancelet Branchiostoma caribaeum Sandevall, 1853 was studied in four surveys, corresponding to austral seasons, in a tropical bay, southeast of Brazil. Abundance was higher in the spring and was positively correlated to coarse sediments, limiting its occurrence to some sectors of the sampling area. Body length and biomass differed seasonally but not between sexes. Sexually mature individuals occurred in all seasons, suggesting continuous breeding that is typical of tropical species. Variation in the frequency of small specimens indicates temporal differences in the intensity of breeding. The body length of recruits differed from other population of lancelets and the small length which B. caribaeum attained sexual maturity in Guanabara Bay may be related to local environmental stress or the great availability of food.

Características clínico-epidemiológicas da doença Lyme-símile em crianças

OBJETIVO: Determinar a prevalência, distribuição etária, sazonalidade, características clínicas da doença Lyme-símile em menores de 15 anos. MÉTODOS: De julho/1998 a dezembro/2000 foi conduzido um estudo transversal em 333 pacientes, com exantema e febre. Foram coletadas amostras pareadas de sangue para a identificação de patógenos. Somente em 193 amostras, negativas aos outros patógenos (Parvovirus B19, Herpesvírus 6 humano, Sarampo, Rubéola, Dengue, Escarlatina e Enterovírus), foram realizadas a pesquisa da borreliose pelos métodos de Enzyme-Linked Immunosorbent Assay e Western-blotting. Outras variáveis clínicas, socioeconômicas, demográficas e climáticas foram estudadas. RESULTADOS: A prevalência da doença foi de 6,2%(12/193). Das variáveis estudadas, houve predomínio em <6anos(83,2%); sexo feminino (66,7%); procedência da cidade de Franco da Rocha (58,3%); com sazonalidade no outono-verão. O intervalo de atendimento foi de quatro dias. Sinais e sintomas com significância estatística: prurido, ausência da fissura labial e bom estado clínico. Outros dados presentes foram: irritabilidade (80%); febre (?38ºC) (58,3%) com duração de um a três dias. O exantema foi do tipo máculo-papular (33,3%), urticariforme (25%) e escarlatiniforme (16,7%); predominando em tronco (60%). Não houve apresentação clínica característica para diagnóstico da doença de Lyme-símile nestes pacientes. A sensibilidade e especificidade para o diagnóstico clínico contraposta com o diagnóstico laboratorial foi zero. O acompanhamento de 10 casos durante dois anos não evidenciou complicações cardiológicas ou neurológicas. Este é o primeiro estudo desta doença em crianças brasileiras. CONCLUSÃO: A prevalência da doença Lyme-símile foi baixa, não tendo sido lembrada no diagnóstico inicial dos exantemas, mas seu conhecimento é necessário, necessitando maior atenção médica.

Transtornos psiquiátricos no pós-parto

OBJETIVO: O pós-parto é um período de alterações biológicas, psicológicas e sociais. Essa é considerada a época mais vulnerável para a ocorrência de transtornos psiquiátricos. A disforia puerperal, a depressão pós-parto e a psicose pós-parto têm sido classicamente relacionadas ao pós-parto. Atualmente, tem sido observado que os transtornos ansiosos também estão associados a esse período. MÉTODO: Neste artigo é feita uma revisão da bibliografia acerca de transtornos psiquiátricos no pós-parto a partir de artigos encontrados no PubMed e no SciELO entre os anos de 2000 e 2009. Livros, teses e outros artigos considerados relevantes citados no material consultado também foram incluídos. RESULTADOS: A disforia puerperal ocorre em 50% a 85% das mulheres, o quadro é leve e transitório e não requer tratamento. A depressão pós-parto tem prevalência em torno de 13%, pode causar repercussões negativas na interação mãe-bebê e em outros aspectos da vida da mulher e deve ser tratada. A psicose pós-parto é rara, aparecendo em cerca de 0,2% das puérperas. Tem quadro grave que envolve sintomas psicóticos e afetivos, havendo risco de suicídio e infanticídio e geralmente requerendo internação hospitalar. Os transtornos ansiosos podem ser exacerbados ou precipitados no pós-parto, especialmente o transtorno de ansiedade generalizada, o transtorno de estresse pós-traumático e o transtorno obsessivo-compulsivo. CONCLUSÃO: Apesar de não serem reconhecidos como entidades diagnósticas pelos sistemas classificatórios atuais, os transtornos mentais no puerpério apresentam peculiaridades clínicas que merecem atenção por parte de clínicos e pesquisadores.

Harmonização e classificação toxicológica de agrotóxicos em 1992 no Brasil e a necessidade de prever os impactos da futura implantação do GHS

Em 1992, o Brasil modificou seus critérios de classificação toxicológica de agrotóxicos adequando-os à recomendação de classificação de periculosidade da Organização Mundial da Saúde (OMS). Em 2002, o Sistema Globalmente Harmonizado de Classificação e Rotulagem de Produtos Químicos (GHS) foi adotado pela Organização das Nações Unidas. Em decorrência, a OMS está adequando ao GHS sua recomendação de classificação de agrotóxicos, o que também deverá ser feito pelo Brasil. Considerou-se oportuno estimar o impacto da alteração de critérios, ocorrida em 1992, na reclassificação toxicológica dos produtos comerciais que se encontravam registrados na ocasião. Encontrou-se que 58,6% do total dos agrotóxicos então registrados (74,9% das formulações líquidas e 31,0% das sólidas) podem ter sofrido reclassificação para classes toxicológicas consideradas de me-nor periculosidade, sofrendo mudanças na comunicação de riscos expressa na rotulagem. Isto pode ter ocasionado conseqüências negativas devido a confusões de interpretação pelos agricultores. Nos países que já dispõem de sistemas de classificação de periculosidade de agrotóxicos, como o Brasil, recomenda-se estimar, antes da implantação, os impactos das mudanças que poderão decorrer da adoção do GHS.

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from meat-producing ewes with mastitis

Avaliou-se a sensibilidade antimicrobiana in vitro de 121 cepas de estafilococos coagulase-negativa isolada de leite de ovelhas Santa Inês, aos fármacos: penicilina, amoxicilina, ampicilina, estreptomicina, oxaciclina, neomicina, cefalotina, gentamicina e sulfonamida. A resistência à sulfonamida foi a mais frequente (27,3%), seguida pela estreptomicina (14,0%) e pela oxaciclina (14,0%), enquanto da gentamicina (1,6%) foi a menos frequente. Todas as cepas foram sensíveis a pelo menos um antimicrobiano, e 20,3% das cepas apresentaram resistência múltipla. Os resultados mostram a importância de Staphylococci coagulase-negativas como agentes causadores de mastite em ovinos, e o perfil de resistência múltipla indica a importância da determinação da resistência à oxaciclina como indicador da presença de ilhas de patogenicidade que contêm fatores de virulência e resistência a outros antimicrobianos que contribuem para a sobrevivência da bactéria ao tratamento.