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Item Expression data of FOS and JUN genes and FTIR spectra provide diagnosis of thyroid carcinoma(Elsevier) Queiroz, João Paulo da Silva; Pupin, Breno; Bhattacharjee, Tanmoy Tapobrata; Uno, Miyuki; Chammas, Roger; Kulcsar, Marco Aurélio Vamondes; Canevari, Renata de AzevedoWe explore the feasibility of using FOS and JUN gene expression and ATR-FTIR for diagnosis of thyroid cancer. For the study, 38 samples (6 non-neoplastic (NN), 10 papillary thyroid carcinoma (PTC), 7 follicular thyroid carcinoma (FTC), and 15 benign tumors (BT) were subjected to RNA extraction followed by quantitative real time PCR (qRT-PCR) and 30 samples (5 NN, 9 PTC, 5 FTC, and 11 BT) were used for Attenuated Total Reflectance – Fourier Transform Infrared (ATR-FTIR) followed by multivariate analysis. Of the above, 20 samples were used for both gene expression and ATR-FTIR studies. We found FOS and JUN expression in malignant tumor samples to be significantly lower than NN and benign. ATR-FIR after multivariate analysis could identify the difficult to diagnose FTC with 93 % efficiency. Overall, results suggest the diagnostic potential of molecular biology techniques combined with ATR-FTIR spectroscopy in differentiated thyroid carcinomas (PTC and FTC) and BT.Item Molecular effects of photodynamic therapy with curcumin on Leishmania major promastigotes(Parasitology Research, Springer-Verlag GmbH Germany) Marcolino, Luciana Maria Cortez; Pinto, Juliana Guerra; Ferreira, Isabelle; Godoi, Bruno Henrique; Canevari, Renata de Azevedo; Ferreira-Strixino, JulianaLeishmaniasis is a neglected disease mainly affecting low-income populations. Conventional treatment involves several side effects, is expensive, and, in addition, protozoa can develop resistance. Photodynamic therapy (PDT) is a promising alternative in treating the disease. PDT involves applying light at a specific wavelength to activate a photosensitive compound (photosensitizer, PS), to produce reactive oxygen species (ROS). Curcumin and its photochemical characteristics make it a good candidate for photodynamic therapy. Studies evaluating gene expression can help to understand the molecular events involved in the cell death caused by PDT. In the present study, RNA was extracted from promastigotes from the control and treated groups after applying PDT. RT-qPCR was performed to verify the expression of the putative ATPase beta subunit (ATPS), ATP synthase subunit A (F0F1), argininosuccinate synthase 1 (ASS), ATP-binding cassette subfamily G member 2 (ABCG2), glycoprotein 63 (GP63), superoxide dismutase (FeSODA), and glucose-6-phosphate dehydrogenase (G6PDH) genes (QR). The results suggest that PDT altered the expression of genes that participate in oxidative stress and cell death pathways, such as ATPS, FeSODA, and G6PD. The ATP-F0F1, ASS, and GP63 genes did not have their expression altered. However, it is essential to highlight that other genes may be involved in the molecular mechanisms of oxidative stress and, consequently, in the death of parasites.Item Busca de marcadores moleculares preditivos do acometimento de linfonodos em carcinomas papilíferos de tireoide(Universidade do Vale do Paraíba - UNIVAP) Oliveira, Maria Luiza Lopes de; Costa, João Vitor Alcântara da; Silva, Luana Araujo da; Santos, Joyce Nascimento; Canevari, Renata de AzevedoO câncer de tireoide é a neoplasia que mais cresce em incidência todos os anos, sendo o carcinoma papilífero de tireoide (CPT) o subtipo mais comum. Em muitos casos estes tumores podem se tornar agressivos, ocasionando metástases linfonodais. A determinação de marcadores moleculares pela análise de expressão gênica é uma importante ferramenta para a determinação do prognóstico destes tumores. Considerando o papel dos genes NEDD9, B3GNT7, PHB, BAD, PAXIP1, PPM1D e PIK3R5 na etiologia de diversos cânceres, este estudo consistiu em verificar pela técnica de RT-qPCR se os mesmos podem ser marcadores moleculares preditivos do acometimento de linfonodos e se estão relacionados ao desenvolvimento destes tumores. No presente estudo não foi observada expressão diferencial significativa dos genes analisados entre os grupos amostrais de CPT versus MCPT (microcarcinomas) e entre os grupos de CPT linfonodo positivo versus CPT linfonodo negativo, indicando que estes genes não estão relacionados ao desenvolvimento tumoral e não podem ser considerados marcadores preditivos de metástases em linfonodos em CPT.Item PCR-RFLP and FTIR-based detection of high-risk human papilloma virus for cervical cancer screening and prevention(Elsevier) Melo, Igor Martins Alves; Viana, Magda Rogéria Pereira; Pupin, Breno; Bhattacharjee, Tanmoy Tapobrata; Canevari, Renata de AzevedoBackground: Approximately 70% of cervical carcinoma cases show the presence of high-risk Human Papilloma Virus (HPV), especially HPV-16 and HPV-18, and can be used to stratify high risk patients from low risk and healthy. Currently, molecular biology techniques such as polymerase chain reaction (PCR) are used to identify the presence of virus in patient samples. While the methodology is highly sensitive, it is labor intensive and timeconsuming. Alternative techniques, such as vibrational spectroscopy, has been suggested as a possible rapid alternative. Therefore, in this study, we evaluate the efficiency of cervical fluid Fourier Transform Infrared spectroscopy (FTIR) in patient risk stratification informed by PCR. Methods: Cervical fluid samples (n = 91) were obtained from patients who have undergone routine Papanicolaou (Pap) test. Viral genome was identified and classified as high/low-risk by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP). FTIR spectra were acquired from samples identified by PCR-RFLP as No-HPV (n = 10), high-risk HPV (n = 7), and low-risk HPV (n = 7). Results: Of the 91 samples, was detected the viral genome by PCR in 36 samples. Of these 36 samples, nine samples were identified to contain high-risk HPV (HR-HPV) and nine samples were found to have low-risk HPV (LR-HPV). The FTIR spectra acquired from No-HPV, LR-HPV, and HR-HPV showed differences in 1069, 1437, 1555, 1647, 2840, 2919, and 3287 cm-1 bands. Principal Component Analysis (PCA) showed distinct clusters for No-HPV and HR-HPV and No-HPV and LR-HPV, but there was significant overlap in the clusters of HR-HPV and LR-HPV. PCA-Linear Discriminant Analysis (PC-LDA) after Leave One Out Cross Validation (LOOCV) classified No-HPV from HR-HPV and No-HPV from LR-HPV with 100% efficiency in the 1400-1800 cm-1 spectral range. LOOCV classifications for LR-HPV and HR-HPV from each other were 71 and 75%, respectively, in the 2800-3400 cm-1 spectral range. Conclusions: The results highlight the high sensitivity of PCR-RFLP in HPV identification and show that FTIR can classify samples identified as healthy, low, and high-risk samples by PCR-RFLP. General significance: We show the possibility of using FTIR for initial cervical cancer risk stratification followed by detailed PCR-RFLP investigations for suspect cases.