Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens

Trypanosoma cruzi trypomastigotes excrete-secrete a complex mixture of antigenic molecules. This antigenic mixture denominated trypomastigote excreted-secreted antigens contains a 150-160 kDa band that shows excellent performance in Chagas' disease diagnosis by immunoblotting. The present study partially characterized by two-dimensional gel electrophoresis the immunoreactivity against the 150-160kDa protein using sera samples from chagasic patients in different phases of the disease. Trypomastigote excreted-secreted antigen preparations were subjected to high-resolution two-dimensional (2D) gel electrophoresis followed by immunoblotting with sera from chagasic and non-chagasic patients. The 150-160kDa protein presented four isoforms with isoelectric focusing ranging from 6.2 to 6.7. The four isoforms were recognized by IgM from acute phase and IgG from chronic phase sera of chagasic patients. The 150-160kDa isoform with IF of approximately 6.4 became the immunodominant spot with the progression of the disease. No cross-reactivity was observed with non-chagasic or patients infected with Leishmania sp. In this study we provide basic knowledge that supports the validation of trypomastigote excreted-secreted antigens for serological diagnosis of Chagas' disease.

Semantics bias in cross-national comparative analyses : Is it good or bad to have "fair" health?

The Health Behavior in School-aged Children is a cross-national study collecting data on social and health indicators on adolescents in 43 countries. The study provides comparable data on health behaviors and health outcomes through the use of a common protocol, which have been a back bone of the study sine its initiation in 1983. Recent years, researchers within the study have noticed a questionable comparability on the widely used item on self-rated health. One of the four response categories to the item "Would you say your health is….?" showed particular variation, as the response category "Fair" varied from 20 % in Latvia and Moldova to 3-4 % in Bulgaria and Macedonia. A qualitative mini-survey of the back-translations showed that the response category "Fair" had a negative slant in 25 countries, a positive slant in 10 countries and was considered neutral in 9 countries. This finding indicates that there are what may be called semantic issues affecting comparability in international studies, since the same original word (in an English original) is interpreted differently across countries and cultures. The paper test and discuss a few possible explanations to this, however, only leaving to future studies to hold a cautious approach to international comparisons if working with the self-rated health item with four response categories.

Behaviour of a dovetail joint solved using force analyses

Dovetail joints are one of the most cornrnonly used joints during Gothic and Baroque periods. Despite being coristructed in the sarne way during the ages, there is no analyticai solution available to help its analysis and design so required in reconstruction works of existing timber structures. An analytical solution based on the principie of virtual works under different types of loading is presented in this paper as weli as differences in bearing capacity of the joint for chosen types of loading, when different wood species are used. The accuracy of the presented solution is confirrned by data obtained from experimental tests of scaied modeis of a dovetail joint.

Development of a thermoelectric generator for the exhaust of a vehicle

Dissertação de mestrado integrado em Engenharia Mecânica

Microscopy- or DNA-based analyses: Which methodology gives a truer picture of stream-dwelling decomposer fungal diversity?

We assessed aquatic hyphomycete diversity in autumn and spring on oak leaves decomposing in five streams along a gradient of eutrophication in the Northwest of Portugal. Diversity was assessed through microscopy-based (identification by spore morphology) and DNA-based techniques (Denaturing Gradient Gel Electrophoresis and 454 pyrosequencing). Pyrosequencing revealed five times greater diversity than DGGE. About 21% of all aquatic hyphomycete species were exclusively detected by pyrosequencing and 26% exclusively by spore identification. In some streams, more than half of the recorded species would have remained undetected if we had relied only on spore identification. Nevertheless, in spring aquatic hyphomycete diversity was higher based on spore identification, probably because many species occurring in this season are not yet connected to ITS barcodes in genetic databases. Pyrosequencing was a powerful tool for revealing aquatic hyphomycete diversity on decomposing plant litter in streams and we strongly encourage researchers to continue the effort in barcoding fungal species.

Flow cytometry analyses of calcification propensity

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Critical analyses of the introduction of liquid-based cytology in a public health service of the state of São Paulo, Brazil

OBJECTIVE The aim of this study was to compare the performance of the current conventional Pap smear with liquid-based cytology (LBC) preparations. STUDY DESIGN Women routinely undergoing their cytopathological and histopathological examinations at Fundação Oncocentro de São Paulo (FOSP) were recruited for LBC. Conventional smears were analyzed from women from other areas of the State of São Paulo with similar sociodemographic characteristics. RESULTS A total of 218,594 cases were analyzed, consisting of 206,999 conventional smears and 11,595 LBC. Among the conventional smears, 3.0% were of unsatisfactory preparation; conversely, unsatisfactory LBC preparations accounted for 0.3%. The ASC-H (atypical squamous cells - cannot exclude high-grade squamous intraepithelial lesion) frequency did not demonstrate any differences between the two methods. In contrast, the incidence of ASC-US (atypical squamous cells of undetermined significance) was almost twice as frequent between LBC and conventional smears, at 2.9 versus 1.6%, respectively. An equal percentage of high-grade squamous intraepithelial lesions were observed for the two methods, but not for low-grade squamous intraepithelial lesions, which were more significantly observed in LBC preparations than in conventional smears (2.2 vs. 0.7%). The index of positivity was importantly enhanced from 3.0% (conventional smears) to 5.7% (LBC). CONCLUSIONS LBC performed better than conventional smears, and we are truly confident that LBC can improve public health strategies aimed at reducing cervical lesions through prevention programs.

Medida da aliança parental: validação portuguesa e construção de uma versão reduzida

Objetivo: A Medida de Aliança Parental (PAM) avalia a qualidade da relação interparental na prestação de cuidados da criança. O presente artigo apresenta a validação de uma versão portuguesa da medida, bem como examina as qualidades psicométricas de uma versão reduzida com 6 itens do instrumento (PAM-R). Método: A amostra foi constituída por 182 pais (63% mães), dos quais 72 preencheram um instrumento de avaliação dos problemas de ajustamento psicológico das crianças. Resultados: As análises fatoriais confirmatórias não corroboraram a estrutura dos dois modelos testados. No entanto, excelentes valores foram encontrados nos índices de adequação do modelo da PAM-R. Não foram encontrados erros de especificação no modelo unidimensional testado, o que suporta a validade fatorial da versão reduzida da PAM. A PAM-R apresentou excelentes valores de consistência interna e uma correlação negativa e significativa com a medida de problemas de ajustamento das crianças. Conclusões: PAM-R emerge como uma medida que possibilita a avaliação do impacto das dimensões familiares no funcionamento e desenvolvimento psicológico das crianças, em contextos de prestação de cuidados de saúde primários.

Initial diagnostic errors in children suspected of having heart disease: prevalence and long-term consequences

OBJECTIVE: To access the incidence of diagnostic errors in the initial evaluation of children with cardiac murmurs. METHODS: We evaluated our 7-years of experience in a public pediatric cardiology outpatient clinic. Of 3692 patients who were sent to the hospital, 2603 presented with a heart murmur and were investigated. Patients for whom a disagreement existed between the initial and final diagnoses were divided into the following 2 groups: G1 (n=17) with an initial diagnosis of an innocent murmur and a final diagnosis of cardiopathy, and G2 (n=161) with an initial diagnosis of cardiopathy and a final diagnosis of a normal heart. RESULTS: In G1, the great majority of patients had cardiac defects with mild hemodynamic repercussions, such as small ventricular septal defect and mild pulmonary stenosis. In G2, the great majority of structural defects were interventricular communication, atrial septal defect and pulmonary valve stenosis. CONCLUSION: A global analysis demonstrated that diagnostic error in the initial evaluation of children with cardiac murmurs is real, reaching approximately 6% of cases. The majority of these misdiagnoses were in patients with an initial diagnosis of cardiopathy, which was not confirmed through later complementary examinations. Clinical cardiovascular examination is an excellent resource in the evaluation of children suspected of having cardiopathy. Immediate outpatient discharge of children with an initial diagnosis of an innocent heart murmur seems to be a suitable approach.

Errores en sistemas de procesamiento de datos debido a eventos transitorios en interfaces analógicas: aportes a la mitigación de los mismos. Errors in data processing systems due to single transient events affecting analog interfaces: contributions to their mitigation.

Los eventos transitorios únicos analógicos (ASET, Analog Single Event Transient) se producen debido a la interacción de un ión pesado o un protón de alta energía con un dispositivo sensible de un circuito analógico. La interacción del ión con un transistor bipolar o de efecto de campo MOS induce pares electrón-hueco que provocan picos que pueden propagarse a la salida del componente analógico provocando transitorios que pueden inducir fallas en el nivel sistema. Los problemas más graves debido a este tipo de fenómeno se dan en el medioambiente espacial, muy rico en iones pesados. Casos típicos los constituyen las computadoras de a bordo de satélites y otros artefactos espaciales. Sin embargo, y debido a la continua contracción de dimensiones de los transistores (que trae aparejado un aumento de sensibilidad), este fenómeno ha comenzado a observarse a nivel del mar, provocado fundamentalmente por el impacto de neutrones atmosféricos. Estos efectos pueden provocar severos problemas a los sistemas informáticos con interfaces analógicas desde las que obtienen datos para el procesamiento y se han convertido en uno de los problemas más graves a los que tienen que hacer frente los diseñadores de sistemas de alta escala de integración. Casos típicos son los Sistemas en Chip que incluyen módulos de procesamiento de altas prestaciones como las interfaces analógicas.El proyecto persigue como objetivo general estudiar la susceptibilidad de sistemas informáticos a ASETs en sus secciones analógicas, proponiendo estrategias para la mitigación de los errores.Como objetivos específicos se pretende: -Proponer nuevos modelos de ASETs basados en simulaciones en el nivel dispositivo y resueltas por el método de elementos finitos.-Utilizar los modelos para identificar las secciones más propensas a producir errores y consecuentemente para ser candidatos a la aplicación de técnicas de endurecimiento a radiaciones.-Utilizar estos modelos para estudiar la naturaleza de los errores producidos en sistemas de procesamiento de datos.-Proponer soluciones novedosas para la mitigación de estos efectos en los mismos circuitos analógicos evitando su propagación a las secciones digitales.-Proponer soluciones para la mitigación de los efectos en el nivel sistema.Para llevar a cabo el proyecto se plantea un procedimiento ascendente para las investigaciones a realizar, comenzando por descripciones en el nivel físico para posteriormente aumentar el nivel de abstracción en el que se encuentra modelado el circuito. Se propone el modelado físico de los dispositivos MOS y su resolución mediante el Método de Elementos Finitos. La inyección de cargas en las zonas sensibles de los modelos permitirá determinar los perfiles de los pulsos de corriente que deben inyectarse en el nivel circuito para emular estos efectos. Estos procedimientos se realizarán para los distintos bloques constructivos de las interfaces analógicas, proponiendo estrategias de mitigación de errores en diferentes niveles.Los resultados esperados del presente proyecto incluyen hardware para detección de errores y tolerancia a este tipo de eventos que permitan aumentar la confiabilidad de sistemas de tratamiento de la información, así como también nuevos datos referentes a efectos de la radiación en semiconductores, nuevos modelos de fallas transitorias que permitan una simulación de estos eventos en el nivel circuito y la determinación de zonas sensibles de interfaces analógicas típicas que deben ser endurecidas para radiación.

Clinical Coronary In-Stent Restenosis Follow-Up after Treatment and Analyses of Clinical Outcomes

Background: Clinical in-stent restenosis (CISR) is the main limitation of coronary angioplasty with stent implantation. Objective: Describe the clinical and angiographic characteristics of CISR and the outcomes over a minimum follow-up of 12 months after its diagnosis and treatment. Methods: We analyzed in 110 consecutive patients with CISR the clinical presentation, angiographic characteristics, treatment and combined primary outcomes (cardiovascular death, nonfatal acute myocardial infarction [AMI]) and combined secondary (unstable angina with hospitalization, target vessel revascularization and target lesion revascularization) during a minimal follow-up of one year. Results: Mean age was 61 ± 11 years (68.2% males). Clinical presentations included acute coronary syndrome (ACS) in 62.7% and proliferative ISR in 34.5%. CISR was treated with implantation of drug-eluting stents (DES) in 36.4%, Bare Metal Stent (BMS) in 23.6%, myocardial revascularization surgery in 18.2%, balloon angioplasty in 15.5% and clinical treatment in 6.4%. During a median follow-up of 19.7 months, the primary outcome occurred in 18 patients, including 6 (5.5%) deaths and 13 (11.8%) AMI events. Twenty-four patients presented a secondary outcome. Predictors of the primary outcome were CISR with DES (HR = 4.36 [1.44–12.85]; p = 0.009) and clinical treatment for CISR (HR = 10.66 [2.53–44.87]; p = 0.001). Treatment of CISR with BMS (HR = 4.08 [1.75–9.48]; p = 0.001) and clinical therapy (HR = 6.29 [1.35–29.38]; p = 0.019) emerged as predictors of a secondary outcome. Conclusion: Patients with CISR present in most cases with ACS and with a high frequency of adverse events during a medium-term follow-up.

Development-oriented refugee assistance : sustainable, efficient and gender-sensitive? ; theoretical and practical analyses based on a case study of the Rhino camp in Uganda

Magdeburg, Univ., Fak. für Humanwiss., Diss., 2012