Characterization of subpopulations (clones and subclones) of the 21 SF strain of Trypanosoma cruzi after long lasting maintenance in the laboratory

Several studies have shown a clonal structure of Trypanosoma cruzi and its possible correlation with the behavioral heterogeneity of the parasite strains. In the present study, the 21 SF strain, that have been maintained in laboratory by successive passages in mice, for more than 15 years, showing a stability of biological and isoenzymic characteristics has been cloned, with the objective of establishing the characters of its clones and subclones. With the technique of isolation of a single parasite from the blood of infected mice, 5 clones and 14 subclones have been obtained. After four passages into mice, inoculum of 10(5) was obtained for each clone and subclone and inoculated into mice weighing 10 to 12 g. These were used for the study of the biological behavior of the clones: evolution of parasitemia, morphology of blood forms and host mortality. For isoenzymic characterization, the clones and subclones were analyzed for ALAT, ASAT, GPI and PGM enzymes. Results have shown that the 5 clones and the 14 subclones disclosed a biological behavior similar to the parental strain, with minor variability of the parasitemic profiles and also the same isoenzymic patterns. These results confirm the stability of the 21 SF strain and indicate a clonal homogeneity of its populations. This is compatible with the hypothesis that the T. cruzi strains represent an equilibrium of either homogenous or heterogeneous populations.

Genetic Variability and Microdistribution of Triatoma infestans Genotypes and Trypanosoma cruzi Clones in Arequipa Region (Peru)

The genetic variability of Triatoma infestans and Trypanosoma cruzi populations was studied by isoenzyme analysis in two distinct areas of Arequipa province (Peru); one, Santa Rita de Siguas, being an endemic area for Chagas' disease, the second, Arequipa, recently infected. Analysis of T. infestans genetic variability indicates, (i) temporal stability of genotypes found in Santa Rita de Siguas, (ii) high genetic differences between Arequipa and Santa Rita de Siguas populations suggesting minor contact between them, (iii) multiple origin of the T. infestans population in Arequipa, and (iv) poor dispersal capacity of T. infestans: the panmictic unit could be reduce to a house. Parasite isoenzyme analysis was performed in 29 Peruvian stocks of T. cruzi, mainly isolated from bugs taken in a single locality, Santa Rita de Siguas. The results show, (i) a high genetic polymorphism, (ii) nine different multilocus genotypes were detected and clustered in two different clades, (iii) most of the parasite isolates pertained to one of the clade and were genetically similar to those analyzed 12 years before. This sample allowed the study of the mating system of T. cruzi in strict sympatic conditions and gave more strength to the hypothesis of the clonal structure of T. cruzi populations

Contrasting Genomic Variability between Clones from Field Isolates and Laboratory Populations of Schistosoma mansoni

The extent of genomic variability of clones of Schistosoma mansoni obtained from field isolates was compared with that of strains that have been laboratory maintained. Analysis was undertaken using randomly amplified polymorphic dNAs (RAPDs) generated with three primers. Phenograms showing the similarity among the clones were constructed. The data showed that while the laboratory strain is highly homogeneous the clones derived from the field populations were highly variable with 43% of RAPDs exhibiting polymorphisms among 23 clones. Clones isolated from the same infected individual were always more closely grouped than clones from different individuals. The data clearly demonstrated that earlier analyses of the genomic variability in S. mansoni have underestimated this phenomenon due to the failure to examine field isolates

Comparative Analysis by Polymerase Chain Reaction Amplified Minicircles of Kinetoplast DNA of a Stable Strain of Trypanosoma cruzi from São Felipe, Bahia, its Clones and Subclones: Possibility of Predominance of a Principal Clone in this Area

Molecular characterization of one stable strain of Trypanosoma cruzi, the 21 SF, representative of the pattern of strains isolated from the endemic area of São Felipe, State of Bahia, Brazil, maintained for 15 years in laboratory by serial passages in mice and classified as biodeme Type II and zymodeme 2 has been investigated. The kinetoplast DNA (kDNA) of parental strain, 5 clones and 14 subclones were analyzed. Schizodeme was established by comparative study of the fragments obtained from digestion of the 330-bp fragments amplified by polymerase chain reaction (PCR) from the variable regions of the minicicles, and digested by restriction endonucleases Rsa I and Hinf I. Our results show a high percentual of similarity between the restriction fragment lenght polymorphism (RFLP) for the parental strain and its clones and among these individual clones and their subclones at a level of 80 to 100%.This homology indicates a predominance of the same "principal clone" in the 21SF strain and confirms the homogeneity previously observed at biological and isozymic analysis. These results suggest the possibility that the T. cruzi strains with similar biological and isoenzymic patterns, circulating in this endemic area, are representative of one dominant clone. The presence of "principal clones" could be responsible for a predominant tropism of the parasites for specific organs and tissues and this could contribute to the pattern of clinico-pathological manifestations of Chagas's disease in one geographical area.

T cell clones from Schistosoma haematobium infected and exposed individuals lacking distinct cytokine profiles for Th1/Th2 polarisation

T cell clones were derived from peripheral blood mononuclear cells of Schistosoma haematobium infected and uninfected individuals living in an endemic area. The clones were stimulated with S. haematobium worm and egg antigens and purified protein derivative. Attempts were made to classify the T cell clones according to production of the cytokines IL-4, IL-5 and IFN-gamma. All the T cell clones derived were observed to produce cytokines used as markers for the classification of Th1/Th2 subsets. However, the 'signature' cytokines marking each subset were produced at different levels. The classification depended on the dominating cytokine type, which was having either Th0/1 or Th0/2 subsets. The results indicated that no distinct cytokine profiles for polarisation of Th1/Th2 subsets were detected in these S. haematobium infected humans. The balance in the profiles of cytokines marking each subset were related to infection and re-infection status after treatment with praziquantel. In the present study, as judged by the changes in infection status with time, the T cell responses appeared to be less stable and more dynamic, suggesting that small quantitative changes in the balance of the cytokines response could result in either susceptibility or resistant to S. haematobium infection.

Genetic characterization of Trypanosoma cruzi natural clones from the state of Paraíba, Brazil

Eighteen Trypanosoma cruzi stocks from the state of Paraíba, Brazil, isolated from man, wild mammals, and triatomine bugs were studied by multilocus enzyme electrophoresis and random primed amplified polymorphic DNA. Despite the low number of stocks, a notable genetic, genotypic, and phylogenetic diversity was recorded. The presence of the two main phylogenetic subdivisions, T. cruzi I and II, was recorded. The strong linkage disequilibrium observed in the population under survey suggests that T. cruzi undergoes predominant clonal evolution in this area too, although this result should be confirmed by a broader sample. The pattern of clonal variation does not suggests a recent origin by founder effect with a limited number of different genotypes.

Schizodeme analysis of Trypanosoma cruzi Colombian strain clones isolated from the acute phase of murine infection

Colombian strain of Trypanosoma cruzi, biodeme Type III (T. cruzi I), has been cloned by micromanipulation at two phases of the acute infection: early (10 days ) and advanced (30 days). Twelve clones were obtained therefrom. Characterization by their biological and biochemical behavior showed an identity among the several clones and their parental strain, albeit with different degrees of virulence. Molecular characterization of the kinetoplast DNA (kDNA) after amplification by polymerase chain reaction revealed identical profiles of the bands from the kDNA minicircle by the analysis of restriction fragment lenght polymorphism for the isolated clones, their parental strain, and to the clones isolated at two different phases of the infection. Results suggest the predominance of a "principal clone", in the composition of the Colombian strain, responsible for the biological and biochemical behavior. However, no relationship was detected between the molecular profile of kDNA and the degree of virulence presented by the several clones.

Genetic variability in G2 and F2 region between biological clones of human respiratory syncytial virus with or without host immune selection pressure

Human respiratory syncytial virus (HRSV) is an important respiratory pathogens among children between zero-five years old. Host immunity and viral genetic variability are important factors that can make vaccine production difficult. In this work, differences between biological clones of HRSV were detected in clinical samples in the absence and presence of serum collected from children in the convalescent phase of the illness and from their biological mothers. Viral clones were selected by plaque assay in the absence and presence of serum and nucleotide sequences of the G2 and F2 genes of HRSV biological clones were compared. One non-synonymous mutation was found in the F gene (Ile5Asn) in one clone of an HRSV-B sample and one non-synonymous mutation was found in the G gene (Ser291Pro) in four clones of the same HRSV-B sample. Only one of these clones was obtained after treatment with the child's serum. In addition, some synonymous mutations were determined in two clones of the HRSV-A samples. In conclusion, it is possible that minor sequences could be selected by host antibodies contributing to the HRSV evolutionary process, hampering the development of an effective vaccine, since we verify the same codon alteration in absence and presence of human sera in individual clones of BR-85 sample.

Immunological response to re-infections with clones of the Colombian strain of Trypanosoma cruzi with different degrees of virulence: influence on pathological features during chronic infection in mice

Re-infections with Trypanosoma cruzi are an aggravating factor for Chagas disease morbidity. The Colombian strain of T. cruzirepresents multiclonal populations formed by clonally propagating organisms with different tropisms and degrees of virulence. In the present study, the influence of successive inoculations with clones of the Colombian strain, exhibiting different degrees of virulence, on chronic myocarditis and the humoral and cellular immune responses (Col-C1 high virulence, Col-C8 medium virulence and Col-C5 low virulence) were demonstrated. Mice from three groups with a single infection were evaluated during the acute (14th-30th day) and chronic phases for 175 days. An immunofluorescence assay, ELISA and delayed type hypersensitivity (DTH) cutaneous test were also performed. Mice with a triple infection were studied on the 115th-175th days following first inoculation. The levels of IgM and IgG2a were higher in the animals with a triple infection. DTH showed a higher intensity in the inflammatory infiltrate based on the morphometric analysis during a 48 h period of the triple infection and at 24 h with a single infection. The histopathology of the heart demonstrated significant exacerbation of cardiac inflammatory lesions confirmed by the morphometric test. The humoral responses indicate a reaction to the triple infection, even with clones of the same strain.

Elementos para guiar ações visando à orientação empreendedora em organizações de software

Neste artigo, aborda-se a orientação empreendedora (OE) em organizações de software. Tem-se por objetivo propor um conjunto consolidado de elementos que possibilitem guiar ações visando à orientação empreendedora em organizações de software. Para tanto, realizou-se pesquisa de natureza exploratória, com o uso de dados qualitativos. A coleta de dados foi feita por meio de entrevistas em profundidade com executivos de 13 organizações com o intuito de realizar um levantamento de práticas efetivas de orientação empreendedora, para compreensão desse comportamento nas organizações e confirmação dos elementos da base conceitual de orientação empreendedora adotada no estudo. Como resultado, apresenta-se um conjunto de elementos da orientação empreendedora, consolidado a partir da literatura e da prática efetiva das organizações, que constitui importante referência para guiar ações visando à orientação empreendedora em organizações de software. O estudo conclui com o enriquecimento e o detalhamento da base conceitual de orientação empreendedora, com a proposição de categorias, agregação de novos elementos decorrentes da prática organizacional e a consolidação do conjunto de elementos, oferecendo subsídios às organizações de software que desejem desenvolver esse comportamento.

A relação entre a satisfação dos funcionários e a satisfação dos clientes: estudo nas empresas de software do município de Belo Horizonte

A indústria de software tem apresentado crescimento vigoroso nos últimos anos, não só em termos mundiais, como também no Brasil. O segmento é composto basicamente por empresas de pequeno porte, com utilização intensiva de mão de obra especializada. No entanto, estudos indicam que o setor pouco se preocupa com a qualificação de funcionários, talvez porque, normalmente, a demanda supere a oferta, o que relega a um plano secundário as preocupações com a gestão de pessoas, ficando as atenções voltadas para o desenvolvimento e a formatação do serviço propriamente dito. Segundo os modelos teóricos, a participação dos funcionários é de vital importância para o fornecimento de um serviço de qualidade, podendo acarretar a satisfação dos clientes, sua lealdade e, consequentemente, incrementar a rentabilidade da organização. A satisfação dos funcionários pode ser alcançada pela introdução de práticas adequadas de gerenciamento de recursos humanos, incluindo boa comunicação, treinamento e desenvolvimento, trabalho em equipe, incentivo à participação e poder de decisão. Diversos modelos foram propostos para avaliar a satisfação dos clientes, a qual é fruto da qualidade percebida do serviço, de suas expectativas e do valor percebido, de acordo com um dos modelos em questão. Alguns estudos relatam a existência de relação positiva e significativa entre satisfação de clientes e de funcionários. Nesse contexto, procurou-se avaliar qual o impacto da satisfação dos funcionários diretamente sobre a satisfação dos clientes, com base em um modelo teórico que integrasse práticas de Recursos Humanos e antecedentes da satisfação dos clientes. Por meio de pesquisa quantitativa, foram entrevistados clientes e funcionários de 50 empresas da indústria de software de Belo Horizonte (Minas Gerais, Brasil), totalizando uma amostra válida de 147 respondentes para cada grupo. Os dados foram avaliados com o recurso da modelagem de equações estruturais, pelo método dos mínimos quadrados parciais. Com base nos resultados obtidos, infere-se que as melhores práticas de Recursos Humanos realmente influenciam a satisfação dos funcionários e que a qualidade percebida do serviço influencia positivamente a satisfação dos clientes. Contudo, não se constatou relação significativa entre a satisfação dos funcionários e a satisfação dos clientes.

Avaliação de alunos e professores acerca do software "Sinais Vitais"

Este artigo teve por objetivo levantar as opiniões de alunos e professores sobre o software "Sinais Vitais". O trabalho foi desenvolvido no Departamento de Enfermagem da Universidade Federal do Ceará. Trabalhou-se com um total de seis alunos e três professores, os quais foram submetidos a uma entrevista após a utilização do software. As entrevistas geraram dez categorias, as quais foram separadas em dois temas: Características que estimularam a utilização do software "Sinais Vitais" e Software educacionalmente correto. Concluiu-se que os professores valorizaram a correção do conteúdo, enquanto os alunos enfocaram mais a dinâmica do programa.

A construção de um software educativo sobre ausculta dos sons respiratórios

O estudo objetivou relatar a experiência da construção de um software educativo sobre a ausculta dos sons respiratórios. Nesse processo, aplicou-se o referencial pedagógico de Lev Semenovic Vygotsky, e delimitou-se o conteúdo de modo a abarcar o método da ausculta passo a passo, as finalidades, a classificação dos sons normais e adventícios e as respectivas características estetoacústicas. Para o desenvolvimento do sistema, foi utilizada a tecnologia 3D, incluindo avatares (personificações virtuais do paciente, do enfermeiro e dos objetos) e ambiente virtual para a ausculta, usando procedimentos de simulação. Ao final do processo, concluiu-se que iniciativas dessa natureza, embora sejam permeadas de dificuldades, trazem contribuições importantes ao ensino-aprendizagem de conteúdos relativos à área da Enfermagem.

Informática em enfermagem: desenvolvimento de software livre com aplicação assistencial e gerencial

O presente estudo teve como objetivo desenvolver um sistema de informação em enfermagem com aplicação na assistência de enfermagem e no gerenciamento do serviço. O SisEnf - Sistema de Informação em Enfermagem - é um software livre composto pelo módulo assistencial de enfermagem: histórico, exame clínico e plano de cuidados; o módulo gerencial compõe-se de: escala de serviço, gestão de pessoal, indicadores hospitalares e outros elementos. O sistema foi implementado na Clínica Médica do Hospital Universitário Lauro Wanderley, da Universidade Federal da Paraíba. Tendo em vista a necessidade de aproximação entre usuário e desenvolvedor, e a constante mudança de requisitos funcionais durante o processo iterativo, foi adotado o método do processo unificado. O SisEnf foi desenvolvido sobre plataforma WEB e com emprego de software livre. Portanto, o trabalho desenvolvido procurou auxiliar o processo de trabalho da enfermagem que agora terá oportunidade de incorporar a tecnologia da informação na sua rotina de trabalho.