Establishing a cell biology platform: isolation and preservation of human blood products

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates

INTRODUCTION: Candida albicans is responsible for superficial or systemic infections known as candidiasis, which may be found in infected tissue as unicellular budding yeasts, hyphae, or pseudohyphae. In this study, the effects of both fluconazole and itraconazole antifungal agents on the hyphal formation and genotypic characterization of C. albicans isolates classified as either susceptible or resistant were investigated. METHODS: The hyphal production of five C. albicans isolates under the action of antifungal agents was investigated by culturing yeast on growth medium and on hyphal induction medium. The genotypic characterization was carried out for 13 isolates of C. albicans using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method. RESULTS: The dimorphism analysis showed that the hyphal formation was higher in resistant than in the susceptible isolates to both azoles. The RAPD-PCR method identified the formation of two different groups. In group A, four resistant and two susceptible isolates were clustered, and in group B, one resistant and six susceptible isolates were clustered. CONCLUSIONS: Considering that hyphal formation was higher in resistant isolates in the presence of azole drugs, we confirmed that the hyphal production is closely related to susceptibility to azoles. These drugs may affect the morphogenesis of C. albicans depending on their susceptibility to these drugs. In relation to RAPD-PCR, most resistant isolates classified in group A and susceptible isolates in group B demonstrated that this method presented a similar standard between the two groups, suggesting that by this technique, a strong correlation between genotypes and fluconazole-resistant samples may be found.

Epidemiological profile of meningococcal disease in the State of Minas Gerais and in the Central, North, and Triângulo Mineiro regions, Brazil, during 2000-2009

INTRODUCTION: Infection by Neisseria meningitidis, termed as meningococcal disease, can cause meningococcal meningitis and septicemia with or without meningitis. Meningococcal disease is endemic in Brazil and has a high potential to cause large-scale epidemics; therefore, it requires the immediate notification of cases to the Information System for Notifiable Diseases (SINAN) in Brazil. The aim of this study was to describe an epidemiological profile using data from notified and confirmed cases in the State of Minas Gerais, Brazil, from January 2000 to December 2009, obtained from the investigation records of individuals with meningitis registered with SINAN. METHODS: This was a retrospective, population-based study. Descriptive analysis of the data was made using the simple and relative frequencies of the categorical variables in the investigation records. RESULTS: There were 1,688 confirmed patients in Minas Gerais of which 45.5% lived in the Central, North, and Triângulo Mineiro regions. The highest frequencies of cases were in the 1-4-years age group (26.3%), males (54.7%), caucasian (36.4%), and lived in an urban area (80%). In the patients with specified education, 650 (60.9%) patients had secondary education. Serogrouping of meningococci had been performed in 500 (29.6%) patients by age and gender; 285 (57%) belonged to serogroup C, 67 (13.4%) were in the 1-to 4-years age group, and 168 (33.6%) were male. CONCLUSIONS: The epidemiological profiles of patients in the Central, North, and Triângulo Mineiro regions were not significantly different from the profile of patients in Minas Gerais.

Development and validation of gold nanoprobes for human SNP detection towards commercial application

Conventional molecular techniques for detection and characterization of relevant nucleic acid (i.e. DNA) sequences are, nowadays, cumbersome, expensive and with reduced portability. The main objective of this dissertation consisted in the optimization and validation of a fast and low-cost colorimetric nanodiagnostic methodology for the detection of single nucleotide polymorphisms (SNPs). This was done considering SNPs associated to obesity of commercial interest for STAB VIDA, and subsequent evaluation of other clinically relevant targets. Also, integration of this methodology into a microfluidic platform envisaging portability and application on points-of-care (POC) was achieved. To warrant success in pursuing these objectives, the experimental work was divided in four sections: i) genetic association of SNPs to obesity in the Portuguese population; ii) optimization and validation of the non-cross-linking approach for complete genotype characterization of these SNPs; iii) incorporation into a microfluidic platform; and iv) translation to other relevant commercial targets. FTO dbSNP rs#:9939609 carriers had higher body mass index (BMI), total body fat mass, waist perimeter and 2.5 times higher risk to obesity. AuNPs functionalized with thiolated oligonucleotides (Au-nanoprobes) were used via the non-cross-linking to validate a diagnostics approach against the gold standard technique - Sanger Sequencing - with high levels of sensitivity (87.50%) and specificity (91.67%). A proof-of-concept POC microfluidic device was assembled towards incorporation of the molecular detection strategy. In conclusion a successful framework was developed and validated for the detection of SNPs with commercial interest for STAB VIDA, towards future translation into a POC device.

Psychosocial factors associated with adherence to treatment and quality of life in people living with HIV/AIDS in Brazil

Objective: The objective of this article was to investigate the biopsychosocial factors that influence adherence to treatment and the quality of life of individuals who have been successfully following the HIV/AIDS treatment. Methods: It is a cross-sectional study carried out with 120 HIV positive participants in the south of Brazil. Among the variables studied, of note are: perceived stress, social support, symptoms of anxiety and depression and quality of life. Results: The results show that a moderate to high adherence to the treatment paired with a strong sense of social support indicate a higher quality of life. Conclusion: The combination of social support and antiretroviral treatment have an impact on physical conditions, improving immune response and quality of life.

Flora Americae Septentrionalis; or, A systematic arrangement and description of the plants of North America. Containing, besides what have been described by preceding authors, many new and rare species, collected during twelve years travels and residence in that country,

v.1 (1814)

Flora Americae Septentrionalis; or, A systematic arrangement and description of the plants of North America. Containing, besides what have been described by preceding authors, many new and rare species, collected during twelve years travels and residence in that country,

v.2 (1814)

Factores Críticos del Desarrollo Local y Regional Integrado zona serrana y Norte de la Provincia de Córdoba-. Validación causal estadística de Modelos de Ecuaciones Estructurales. Identificación de problemas normativos de las OSC.Critical Factors of Local and Regional Development in north of the Province of Córdoba. Causal validation with Structural Equation Modeling. Legal regulations of civil society.

En la investigación anterior -en la zona pampeana de la Provincia de Córdoba- se demostró teórica y empíricamente, que el desarrollo de la Sociedad Civil muchas veces libradas a su suerte y con limitaciones legales apoyan decididamente el desarrollo local, sin embargo han logrado solo parcialmente sus objetivos, por lo que es necesario comenzar un camino de fortalecimiento en los nuevos roles que deben asumir. Los gobiernos locales, a la vez, intentan trabajosamente con contados éxitos detener el procesos de descapitalización social -financiera y humana- de sus comunidades locales y regionales, peregrinando con escaso éxito a los centros concentrados del poder político y económico, para procurar los recursos financieros y humanos necesarios que no alcanzan a reponer los que se fugan desde hace décadas de sus localidades. Las empresas, con ciclos recurrentes de crecimiento y decrecimiento vinculados a los mercados en que colocan sus productos, también se debaten en la búsqueda de los escasos recursos, financieros y humanos, que les permitan consolidar un desarrollo a mediano y largo plazo. El desarrollo alcanzado en Sistemas de información, instrumentos de relevamiento, análisis y elaboración de propuestas para el Desarrollo Local, nos permite avanzar en: 1. La confirmación empírica de las hipótesis iniciales - factores exógenos y endógenos - en la zona Norte y Serrana de la provincia 2. La validación científica -mediante el Análisis de ecuaciones estructurales. de tales supuestos, para el conjunto de las poblaciones analizadas en ambas etapas. 3. La identificación de los problemas normativos que afectan el desarrollo de las Organizaciones de la Sociedad Civil (OSC). METODOLOGÍA Respecto la validación empírica en la zona norte y serrana 1. Selección de las 4 localidades a relevar de acuerdo a las categorías definidas 2. Elaboración de acuerdos con autoridades e instituciones locales. 3. Relevamiento cualitativo con líderes locales y fuentes de datos secundarias. 4. Adaptación de instrumentos de relevamiento a las realidades locales y estudios previos 5. Relevamiento cuantitativo de campo, capacitación de encuestadores y supervisores. 6. Procesamiento y elaboración de informes finales locales. Respecto de la construcción de modelos de desarrollo 1. Desarrollar las dimensiones especificas y las variables (items) de cada factor crítico. 2. Revisar el instrumento con expertos de cada una de las dimensiones. 3. Validar a nivel exploratorio por medio de un Análisis de Componentes Principales 4. Someter a los expertos la evaluación de una serie de localidades que representan cada uno. Respecto de la identificación de las normas legales que afectan a la Sociedad Civil 1.Relevamiento documental de normas 2. Relevamiento con líderes de instituciones de la Sociedad Civil 3. Análisis de las normas vigentes 4. Elaboración de Informes Finales y Transferencia a líderes e instituciones

Catalogue of birds of the Americas and the adjacent islands in Field Museum of Natural History. including all species and subspecies known to occur in North America, Mexico, Central America, South America, the West Indies, and islands of the Caribbean Sea, the Galapagos Archipelago, and other islands which may properly be included on account of their faunal affinities /

v.13:pt.2:no.1(1918)

The fur seals and fur-seal islands of the North Pacific ocean / By David Starr Jordan, [et al.]

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