Molecular profiling of drug resistant isolates of Mycobacterium tuberculosis in the state of Santa Catarina, southern Brazil.

Submitted by Nuzia Santos ([email protected]) on 2016-02-22T18:00:26Z No. of bitstreams: 1 Molecular profiling of drug resistant isolates of Mycobacterium tuberculosis in the state of Santa Catarina, southern Brazil..pdf: 292609 bytes, checksum: 4bea0aaa468b44fe8ec1bba4d8017a9e (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-02-22T18:06:23Z (GMT) No. of bitstreams: 1 Molecular profiling of drug resistant isolates of Mycobacterium tuberculosis in the state of Santa Catarina, southern Brazil..pdf: 292609 bytes, checksum: 4bea0aaa468b44fe8ec1bba4d8017a9e (MD5)Made available in DSpace on 2016-02-22T18:06:23Z (GMT). No. of bitstreams: 1 Molecular profiling of drug resistant isolates of Mycobacterium tuberculosis in the state of Santa Catarina, southern Brazil..pdf: 292609 bytes, checksum: 4bea0aaa468b44fe8ec1bba4d8017a9e (MD5) Previous issue date: 2015Universidade Federal de Santa Catarina. Laboratório de Biologia Molecular, Sorologia e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular, Sorologia e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular, Sorologia e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular, Sorologia e Micobactérias. Florianópolis, SC, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, BrasilLaboratório Central de Saúde Pública de Santa Catarina. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular, Sorologia e Micobactérias. Florianópolis, SC, BrasilDrug resistance is a global threat and one of the main contributing factors to tuberculosis (TB) outbreaks. The goal of this study was to analyse the molecular profile of multidrug-resistant TB (MDR-TB) in the state of Santa Catarina in southern Brazil. Fifty-three MDR Mycobacterium tuberculosis clinical isolates were analysed by spoligotyping and a partial region of the rpoB gene, which is associated with rifampicin resistance (RMP-R), was sequenced. Some isolates were also distinguished by their mycobacterial interspersed repetitive units (MIRU). S531L was the most prevalent mutation found within rpoB in RMP-R isolates (58.5%), followed by S531W (20.8%). Only two MDR isolates showed no mutations within rpoB. Isolates of the Latin American Mediterranean (LAM) family were the most prevalent (45.3%) found by spoligotyping, followed by Haarlem (9.4%) and T (7.5%) families. SIT106 was found in 26.4% of isolates and all SIT106 isolates typed by MIRU-12 (5 out of 14) belong to MIT251. There was a high correlation between the S531W mutation and the LAM family mainly because all SIT2263 (LAM9) isolates carry this mutation. Among isolates with the S531W mutation in rpoB MIRU demonstrates a cluster formed by four isolates (SIT2263 and MIT163) and very similar profiles were observed between eight of the nine isolates. Better characterisation of TB isolates may lead to new ways in which to control and treat TB in this region of Brazil

RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia.

Submitted by Nuzia Santos ([email protected]) on 2016-01-26T14:59:48Z No. of bitstreams: 1 RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia .pdf: 3813372 bytes, checksum: 8d45353e084f4f7c3069a1cc6c773c95 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-01-26T15:04:58Z (GMT) No. of bitstreams: 1 RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia .pdf: 3813372 bytes, checksum: 8d45353e084f4f7c3069a1cc6c773c95 (MD5)Made available in DSpace on 2016-01-26T15:04:58Z (GMT). No. of bitstreams: 1 RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia .pdf: 3813372 bytes, checksum: 8d45353e084f4f7c3069a1cc6c773c95 (MD5) Previous issue date: 2015Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte MG, BrasilUniversidade Federal de Minas Gerais. Escola de Engenharia. Departamento de Engenharia de Produção. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Exatas. Departamento de Estatística. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Belo Horizonte, MG, BrasilRAP1 (RAS proximate 1), a small GTP-binding protein of the RAS superfamily, is a putative oncogene that is highly expressed in several malignant cell lines and types of cancers, including some types of squamous cell carcinoma. However, the participation of RAP1 in cervical carcinogenesis is unknown. We conducted a cross-sectional study of paraffin-embedded cervical biopsies to determine the association of RAP1 with cervical intraepithelial neoplasia (CIN). Standard and quantitative immunohistochemistry assessment of RAP1 expression in fixed tissue was performed on 183 paraffin-embedded cervical biopsies that were classified as normal or non-dysplastic mucosa (NDM) (n = 33); CIN grade 1 (n = 84) and CIN grade 2/3 (n = 66). A gradual increase in RAP1 expression in NDM < CIN 1 < CIN 2/3 (p<0.001) specimens was observed and was in agreement with the histopathologic diagnosis. A progressive increase in the RAP1 expression levels increased the risk of CIN 1 [odds ratio (OR) = 3.50; 95% confidence interval (CI) 1.30-10.64] 3.5 fold and the risk of CIN 2/3 (OR = 19.86, 95% CI 6.40-70.79) nearly 20 fold when compared to NDM. In addition, stereotype ordinal regression analysis showed that this progressive increase in RAP1 expression more strongly impacted CIN 2/3 than CIN 1. Our findings suggest that RAP1 may be a useful biomarker for the diagnosis of CI

RAP1 GTPase Overexpression is Associated with Cervical Intraepithelial Neoplasia

<div><p>RAP1 (RAS proximate 1), a small GTP-binding protein of the <i>RAS</i> superfamily, is a putative oncogene that is highly expressed in several malignant cell lines and types of cancers, including some types of squamous cell carcinoma. However, the participation of RAP1 in cervical carcinogenesis is unknown. We conducted a cross-sectional study of paraffin-embedded cervical biopsies to determine the association of RAP1 with cervical intraepithelial neoplasia (CIN). Standard and quantitative immunohistochemistry assessment of RAP1 expression in fixed tissue was performed on 183 paraffin-embedded cervical biopsies that were classified as normal or non-dysplastic mucosa (NDM) (n = 33); CIN grade 1 (n = 84) and CIN grade 2/3 (n = 66). A gradual increase in RAP1 expression in NDM < CIN 1 < CIN 2/3 (<i>p<0</i>.<i>001</i>) specimens was observed and was in agreement with the histopathologic diagnosis. A progressive increase in the RAP1 expression levels increased the risk of CIN 1 [odds ratio (OR) = 3.50; 95% confidence interval (CI) 1.30-10.64] 3.5 fold and the risk of CIN 2/3 (OR = 19.86, 95% CI 6.40-70.79) nearly 20 fold when compared to NDM. In addition, stereotype ordinal regression analysis showed that this progressive increase in RAP1 expression more strongly impacted CIN 2/3 than CIN 1. Our findings suggest that RAP1 may be a useful biomarker for the diagnosis of CIN.</p></div

Immunohistochemical staining of RAP1.

<p>Left—Squamocolumnar junction of non-dysplastic cervical mucosa (NDM) showing very weak immunostaining for RAP1 in the cytoplasm of basal cells and no staining in the intermediate and superficial cells. Middle—CIN 1 showing moderate staining for RAP1 in cells distributed in the basal third of the epithelium and showing RAP1 expression in neutrophils. Right- CIN 2/3 showing strong staining for RAP1 in cells diffusely distributed throughout the entire epithelial thickness, with nuclear translocation of RAP1.</p

Morphometric analysis of RAP1 and p16^INK4A immunohistochemical staining.

<p>Relative Intensity (RI) data of RAP1 anfd p16^INK4A expression in NDM, CIN 1, and CIN 2/3 samples are expressed in box plot format, with boxes stretching from the 25^th percentile to the 75^th percentile and the line across the box representing the median values. Statistically significant differences (p <0.05) are shown by dotted lines.</p

Estimated coefficients, standard errors, z-scores, and two-tailed <i>p</i>-values for the fitted stereotype regression model.

<p>Log-likelihood: -136.46</p><p>Abbreviations: SE—Standard error; NDM—non-dysplastic mucosa; CIN—cervical intraepithelial neoplasia.</p><p>* Identifiability constraints proposed proposed by Anderson [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123531#pone.0123531.ref026" target="_blank">26</a>].</p><p>Estimated coefficients, standard errors, z-scores, and two-tailed <i>p</i>-values for the fitted stereotype regression model.</p

Scores of RAP1 and p16^INK4A immunohistochemical expression in precursor lesions of cervical neoplasia.

<p>Abbreviations: NDM—non-dysplastic mucosa; CIN—cervical intraepithelial neoplasia.</p><p>Scores of RAP1 and p16^INK4A immunohistochemical expression in precursor lesions of cervical neoplasia.</p

Associations between RAP1 expression and risk of CIN progression.

<p>Abbreviations: OR—Odds ratio; NDM—non-dysplastic mucosa; CIN—cervical intraepithelial neoplasia.</p><p>Associations between RAP1 expression and risk of CIN progression.</p