Optimization Protocol for the SSRL In-Situ Crystallization and Automated Data Collection Plate

Typically when researchers want to collect data from protein crystals, those crystals must be flash-frozen using liquid nitrogen. This keeps the crystals stable for data analysis, however, can damage crystals and does not give the best possible data. Ideally, crystals can be analyzed at room temperature, which would allow for more modulation of the shape and give researchers a better picture of the shape of the protein. Room temperature data analysis can be difficult, because protein crystals are fragile and will dehydrate quickly if not kept in the proper conditions. The purpose of this research was to develop and optimize a plate that could store crystals at room temperature without damaging the crystals. Over the course of the project many different conditions were tested to try to find an appropriate environment for the protein crystals inside the SSRL In-Situ Crystallization Plate that would allow crystals to remain viable at room temperature for an extended period of time. It was found that if the crystals were loaded into a plate that had been equilibrated with a crystallization solution thickened with agarose, the crystals would remain viable for at least three days. Diffraction patterns were taken from various crystals after one, two, and three days in order to confirm the crystals were indeed viable and that they refracted properly. The end goal of this project is to allow users to SLAC to be able to utilize this plate and the Stanford Automated Mounter located at SSRL to collect data from room temperature crystals remotely without damage to those crystals

A Influência dos Valores Organizacionais no Desempenho de Agências Bancárias

This paper aimed to investigate the influence organizational values at bank branches have on bank branchperformance. The research was undertaken at branches of a Brazilian publicly held bank. The questionnairecalled Inventário de Perfis de Valores Organizacionais (IPVO, or Inventory of Organizational Values Profiles)was adapted to measure the organizational values of bank branches. Bank branch performance was measuredobjectively, using the percentage of goals accomplished during the year 2009. The sample is non-probabilisticand is formed by 271 bank branches and the data about the organizational values of bank branches was providedby 720 respondents. Data analysis revealed that the organizational values of bank branches are organized intofour factors: respect for stakeholders, employee entertainment, competent performance, and prestige. Prestige ispositively correlated with bank branch performance, and respect for stakeholders and employee entertainmentare negatively correlated with the dependent variable. The multiple regression analysis yielded an equation thataccounts for 9.5% of the variance in performance and contains two factors as independent variables whichpositively influence bank branch performance: prestige and competent performance. The results showed thatorganizational values are one of the variables that can impact bank branch performance

Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus

Background: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. Objective: To validate our in-house indirect ELISAgp90/45, following the World Organization of Animal Health (OIE) criteria. Study design: Test validation. Methods: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45. Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. Results: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Main limitations: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. Conclusion: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.Fil: Russi, Romina Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; ArgentinaFil: García, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; ArgentinaFil: Cámara, María Silvia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Soutullo, Adriana Rosa. Gobierno de la Provincia de Santa Fe. Ministerio de la Producción, Ciencia y Tecnología. Laboratorio de Diagnóstico e Investigaciones Agropecuarias; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; Argentin

Assessment of total hepatitis C virus (HCV) core protein in HCV-related mixed cryoglobulinemia

INTRODUCTION: In hepatitis C virus (HCV)-related mixed cryoglobulinemia (MCG), the nonenveloped HCV core protein (HCV-Cp) is a constituent of the characteristic cold-precipitating immune complexes (ICs). A possible correlation between HCV-Cp, virologic, laboratory, and clinical parameters in both untreated MCG patients and those undergoing specific treatment was explored. METHODS: HCV-Cp was quantified by a fully automated immune assay. Correlations between HCV-Cp and HCV RNA, cryocrit, and virus genotype (gt) were investigated in 102 chronically HCV-infected MCG patients. RESULTS: HCV-Cp concentrations strongly correlated with HCV RNA levels in baseline samples. An average ratio of 1,425 IU and 12,850 IU HCV RNA per picogram HCV-Cp was estimated in HCV gt-1 and gt-2 patients, respectively. This equation allowed us to estimate that, on average, HCV-Cp was associated with the viral genome in only 3.4% of the former and in 35% of the latter group of patients. The direct relation between HCV-Cp and the cryocrit level suggests that the protein directly influences the amount of cryoprecipitate. Although the therapy with rituximab (RTX) as a single agent resulted in the enhancement of HCV-Cp levels, in patients treated with RTX in combination with a specific antiviral therapy (pegylated interferon-α plus ribavirin), the prompt and effective clearance of HCV-Cp was documented. CONCLUSIONS: Our data provide evidence that HCV-Cp has a direct effect on the cold-precipitation process in a virus genotype-dependence in HCV-related MCG patients

A mudança organizacional sob a ótica do discurso: um caso de ensino de uma comunidade de pessoas com e sem deficiências de aprendizado

This article presents a teaching case focused on the discussion of organizational change, from the perspective of the interpretive paradigm and of the studies of organizational discourse. It describes a process of change within a non-profit organization, where people with and without learning disabilities, the latter called assistants, live in community. The introduction of new government guidelines changed the system of financing of such activity, giving more autonomy to people with learning difficulties and failing to provide activities that were central to the organization’s system of meanings. With the “heart” of the community threatened, assistants express the challenges to experience organizational change, reporting that they are dancing in blood. The case aims to contribute to the discussion of organizational change processes in postgraduate and possibly undergraduate courses, through an unusual perspective, which states the change not only as deliberate, but mainly as something built in participants’ interactions. It also can be used for supporting the discussion between agents and recipients of change in organizations.Este artigo apresenta um caso de ensino voltado à discussão da mudança organizacional, sob a ótica do paradigma interpretativista e dos estudos dos discursos organizacionais. Descreve-se o processo de mudança pelo qual está passando uma organização não governamental, em que pessoas com e sem deficiências de aprendizado, estes últimos denominados assistentes, vivem em comunidade. A introdução de diretrizes governamentais alterou o sistema de financiamento de tal atividade, dando maior autonomia para as pessoas com deficiências de aprendizado e deixando de prover atividades que eram centrais no sistema de significados da organização. Com o “coração” da Comunidade ameaçado, os assistentes expressam os desafios para vivenciar a mudança organizacional, reportando estarem dançando sobre o sangue. Este estudo visa, assim, contribuir para a discussão de processos de mudança organizacional em cursos de pós-graduação e, eventualmente, de graduação, mediante uma perspectiva ainda pouco empregada, que revela a mudança como um processo não somente deliberado, mas também construído nas interações dos participantes. Além disso, serve para apoiar a discussão entre agentes e recipientes de mudanças nas organizações

Gestores brasileiros e portugueses: Estrutura única de valores de sentido de vida?

Compreender o sentido da vida e os diferentes sentidos que os indivíduos atribuem a suas vidas são desafios que acompanham o ser humano desde os primórdios, auxiliando-o a lidar com perplexidades. Sentidos de vida podem ser considerados valores, uma vez que orientam a conduta humana, passíveis de aprendizado e compartilhamento. Este artigo teve como objetivo comparar a estrutura de valores relativos a sentido de vida de gestores brasileiros e portugueses. Teve como propósito, aprofundar a compreensão de semelhanças e diferenças entre gestores de duas culturas – brasileira e portuguesa – que possuem histórias imbricadas, no entanto, com identidades próprias. De natureza exploratório-descritiva, a pesquisa coletou dados mediante a aplicação da escala de valores de sentido de vida – VSV – a uma amostra de 187 gestores brasileiros e 71 portugueses, tendo os mesmos sido tratados mediante escalonamento multidimensional. Os resultados evidenciaram a existência de uma única estrutura de valores de sentidos de vida para ambas as amostras, no entanto há diferenças de nuances, quanto à composição das categorias nela contempladas, que foram interpretadas tendo em vista as peculiaridades de cada cultura. A estrutura encontrada caracterizou-se como radex, composta de dois círculos concêntricos onde, no externo, situou-se o valor de sentido de vida Evolução Espiritual, e no interno, uma estrutura polar bi-dimensional contemplando as demais categorias teoricamente previstas: Solidariedade Humana versus Auto-realização, Relacionamento com Pessoas Próximas versus Evolução Pessoal.Understanding the meaning of life and the different meanings which individuals attribute to their lives are challenges which accompany the human being from their origins, helping them to deal with perplexities. Meanings of life can be regarded as values, once they orient the human being’s conduct, amenable to learning and sharing. This article was aimed at comparing the structure of values related to the meaning of life of Brazilian and Portuguese managers and to improve the comprehension of the similarities and differences between managers of two cultures – Brazilian and Portuguese – which have intertwined histories although with their own identity. Descriptive-Explanatory in nature, the research collected data through the application of scales of meaning-of-life values – VSV – to a sample of 187 Brazilian and 71 Portuguese managers, dealt with through multidimensional scaling. The results showed the existence of a single structure of meaning-of-life values for both samples, although with some subtle differences, with respect to the composition of the categories contemplated, which were interpreted based on the particularities of each culture. The structure found was made up of a radex composed of two concentric circles, where in the external it was found the meaningof- life value Spiritual Evolution, and in the internal a bi-dimensional polar structure contemplating the other categories theoretically forecasted: Human Solidarity versus Self-fulfilment, Relationship with Close People versus Personal Evolution

Caveolin-1 Expression and Hemodynamics in COPD Patients

Caveolin-1 is a regulator of both intracellular calcium homeostasis and endothelial nitric oxide synthase and may play a pathogenetic role in pulmonary hypertension. In the present study, we aimed to investigate the correlations between pulmonary hemodynamics and vessel morphology including the expression of Caveolin-1 in pulmonary arterioles from patients with chronic obstructive pulmonary disease (COPD) who underwent lung-volume reduction surgery. Staining and subsequent analysis was performed on paraffin-embedded lung tissue from COPD patients (n = 12). Pulmonary arteries with an external diameter of 100-500µm were analysed. Immunhistochemistry with antibodies against caveolin-1 was performed and intensity was assessed. Morphometric data were obtained by using computer-assisted imaging software. The findings were quantified and correlated to hemodynamic data obtained by right-heart catheterization. In COPD patients with pulmonary hypertension (n = 5), the expression of caveolin-1 within the medial smooth muscle cell layer was found to be increased, whereas the intimal caveolin-1 was more prominently expressed in COPD patients with normal pulmonary pressures (n = 7). The ratio between these expression patterns was positively correlated to the mean pulmonary artery pressure. Similar findings were observed for the ratio between intimal and medial thickness as well as for the expression of smooth muscle actin (SMA)

Organizational dignity: a study in a financial organization

Dignidade é um tema abordado em diversos campos de estudo. No âmbito das organizações, discussões têm ocorrido principalmente relacionadas à dignidade no trabalho e do trabalhador. Esforços vêm sendo feitos, no Brasil, para conceituar dignidade no nível organizacional, em um mundo que exige das organizações ações com fins não somente econômicos. Teixeira e colaboradores (2012), considerando a organização como um ator social, vêm desenvolvendo estudos sobre o tema, que culminaram na elaboração de instrumento para mensuração de práticas relativas à dignidade organizacional, aplicado a trabalhadores brasileiros de diversos segmentos econômicos, resultando cinco fatores: práticas de promoção do desenvolvimento participativo do empregado, oferta de produtos e atendimento de qualidade, respeito aos direitos dos empregados, favorecimento do meio social e ambiental e práticas não enganosas com relação aos stakeholders. Neste artigo buscou-se investigar uma organização específica, um banco privado, pesquisando-se as diferenças de percepção entre diferentes grupos sociodemográficos, aplicando-se o instrumento mencionado a uma população de 149 empregados do core business. Os resultados provenientes de tratamento estatístico de 103 casos válidos, usando técnicas paramétricas e não paramétricas, revelaram que práticas voltadas ao desenvolvimento dos empregados e que favorecem o meio ambiente e social não são percebidas como tais, sendo que as mulheres percebem menos as primeiras comparativamente aos homens. A pesquisa comparou, ainda, os resultados dessa amostra com o estudo de Teixeira et al. (2012), apontando diferenças estatisticamente significativas em relação aos cinco fatores, trazendo indícios de que tais características podem variar em função do segmento econômico a que pertencem as empresas.info:eu-repo/semantics/publishedVersio

Aspects of the germination of assa-peixe seeds (Vernonia polyanthes Less.)

O objetivo do presente trabalho foi avaliar o efeito da temperatura e luz na germina??o de sementes de assa-peixe (Vernonia polyanthes Less.). Para tanto foram testadas seis temperaturas constantes (15, 20, 25, 30, 35 e 40?C) e quatro combina??es de temperaturas alternadas (15-20, 15-25, 20-30 e 20-35?C) na presen?a e aus?ncia de luz, a qual foi fornecida durante os per?odos de temperaturas mais altas, quando avaliada a altern?ncia. As vari?veis avaliadas foram: teor de ?gua, germina??o e vigor (primeira contagem e velocidade de germina??o). O delineamento experimental adotado foi o inteiramente casualizado e os resultados foram submetidos ? an?lise da vari?ncia, com as m?dias comparadas pelo teste de Tukey a 5% de probabilidade. Sementes da esp?cie V. polyanthes s?o fotobl?sticas preferenciais, com germina??o favorecida pela presen?a de luz, mas as maiores porcentagens de germina??o e n?veis de vigor dessas sementes foram obtidas em temperaturas de 25?C constante e de 15-25?C alternada, na presen?a de luz.Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG)Empresa de Pesquisa Agropecu?ria de Minas Gerais (EPAMIG)Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq)Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES)The objective of this research was to evaluate the effect of temperature and light on germination of Vernonia polyanthes Less. seeds. Therefore were tested six constant temperatures (15, 20, 25, 30, 35 and 40?C), and four combinations of alternating temperatures (15-20, 15-25, 20-30 and 20-35?C) in the presence and absence, which was provided during periods of higher temperatures, when evaluating the rotation. The variables evaluated were: degree of moisture, germination and vigor (first count of germination and speed of germination). The completely random design was used and the results were submitted to analysis of variance with means compared by Tukey test at 5% probability. Seeds of the Vernonia polyanthes species are preferential photoblastic with germination favored in the presence of light. The highest percentage of germination and seed vigor levels of the seeds of V. polyanthes were obtained in temperatures of 25?C constant and alternating 15-25?C in the presence of light

Structural basis of a histidine-DNA nicking/joining mechanism for gene transfer and promiscuous spread of antibiotic resistance

Relaxases are metal-dependent nucleases that break and join DNA for the initiation and completion of conjugative bacterial gene transfer. Conjugation is the main process through which antibiotic resistance spreads among bacteria, with multidrug-resistant staphylococci and streptococci infections posing major threats to human health. The MOBV family of relaxases accounts for approximately 85% of all relaxases found in Staphylococcus aureus isolates. Here, we present six structures of the MOBV relaxase MobM from the promiscuous plasmid pMV158 in complex with several origin of transfer DNA fragments. A combined structural, biochemical, and computational approach reveals that MobM follows a previously uncharacterized histidine/metal-dependent DNA processing mechanism, which involves the formation of a covalent phosphoramidate histidine-DNA adduct for cell-to-cell transfer. We discuss how the chemical features of the high-energy phosphorus-nitrogen bond shape the dominant position of MOBV histidine relaxases among small promiscuous plasmids and their preference toward Gram-positive bacteria