Navegando por Autor "Pupin, Breno"
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Item Evaluation of hydrogel use in the development of Rapanea ferruginea with water restriction by vibrational Fourier Transform Infrared Spectroscopy (FTIR-UATR)(SciELO) Pereira, Douglas Cubas; Pupin, Breno; Sakane, Kumiko KoibuchiThis study evaluated the use of hydrogel on the development of Rapanea ferruginea under water restriction through Vibrational Fourier Transform Infrared Spectroscopy (FTIR-UATR). Seedlings of approximately 30 cm height were transferred in pots with 3 L of soil. The group of seedlings was separated into 5 different triplicate treatments according to the amount of hydrogel, as follows: H1 (25%), H2 (20%), H3 (15%), H4 (10%) and S (control without hydrogel). The applied hydrogel was mixed homogeneously with the soil. All treatments were irrigated with 40 mL of water. FTIR spectra were obtained from fresh leaves collected during 13 months of monitoring. The correlation of the hydroxyl (water) band with the main biomolecules between treatments with the control was evaluated using the Mann-Whitney test (p<0.05). The biomolecule bands were subjected to principal component analysis (PCA) and hierarchical clustering analysis (HCA). The results indicate a significant correlation of the water band with the lipid, hemicellulose, cell wall and starch components in the species. The use of hydrogel resulted in a significant difference in the water absorption band in relation to the control group through the Mann-Whitney test and in biomolecules as the HCA and PCA analysis suggested. The best development was observed in groups H4 (10%), H2 (20%) and H3 (15%). The use of hydrogel positively influences the biomolecular development of the Rapanea ferruginea and monitoring is viable by FTIR.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 Infrared Spectroscopy Based Study of Biochemical Changes in Saliva during Maximal Progressive Test in Athletes(Springer Nature Link) Vieira, Claudio Augusto Gonçalves de Araujo; Pupin, Breno; Bhattacharjee, Tanmoy T.; Sakane, Kumiko KoibuchiThis study aims to explore biochemical changes in saliva during cardiorespiratory exercise using attenuated-total- reflectance–Fourier-transform-infrared-spectroscopy (ATR-FTIR). Saliva and blood samples were obtained from six athletes at rest, and after running at speeds of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 kilometers-per-hour (km/h) on a treadmill (maximal stress test). Saliva ATR-FTIR spectra were analyzed using deconvolution and multivariate analysis. Area-under-the-curve calculations suggest differential changes in glucose, lactate, protein, lipids, carbohydrate and phosphate content in saliva during the test. Increases in glucose and lactate levels with increasing speeds were verified by simultaneous measurement of blood glucose and lactate levels using standard equipment (Roche®). Multivariate principal-component-analysis (PCA) showed discrete clusters for low (rest-14 km/h) and high (15 – 20 km/h) speeds, and PCA–linear-discriminant-analysis showed 100% classification of 18 – 20 km/h as high speed. Overall, results suggest the possibility of using this non-invasive saliva-based ATR-FTIR method for biochemical assessment during sports exercise and stress tests.Item Molecular Markers for Thyroid Cancer Diagnosis: Insights from MAPK Pathway Gene Expression Analysis(MDPI) Pupin, Breno; Diniz, Ramon Varella; Costa, Maricilia Silva; Chagas, Maurilio José; Santos, André Bandiera de Oliveira; Canevari, Renata de AzevedoBackground and Objectives: Thyroid cancer is the prevailing endocrine malignancy, with incidence growing over the last decades in the world. The current diagnostic techniques often yield inconclusive results, emphasizing the need for more effective diagnostic ap- proaches. Molecular profiling emerges as a promising avenue for carcinoma differentiation, offering precise insights to guide patient selection for surgical intervention. This study aimed to identify molecular markers in thyroid cancer through the expression analysis of genes within the MAPK pathway, aiming to enhance the sensitivity and specificity of carcinoma diagnosis. Methods: Through a comparative analysis of malignant and benign thyroid samples, we identified 46 genes of the MAPK pathway that exhibited differential expression by PCR array analysis. Results: Validation through RT-qPCR and in silico analysis using TCGA confirmed significant results for CCNA1, CDKN1C, CREB1, FOS, HSPA5, JUN, MAP2K6, and SFN genes identified in our cohort, reinforcing the relevance of these biomarkers. Specifically, noteworthy are our findings regarding the potential diag- nostic value of CCNA1 and SFN genes in papillary thyroid carcinoma, while the reduced expression of CDKN1C, FOS, and JUN genes in follicular carcinoma suggests their value in distinguishing the thyroid pathologies. Conclusions: This study identifies promising diagnostic markers, namely CCNA1, CDKN1C, FOS, JUN, and SFN genes, which have the potential to enhance clinical decision-making in thyroid cancer.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.Item Physiological stress tolerance responses of the dung decomposer fungus Mucor circinelloides(Elsevier) Acheampong, Mavis Agyeiwaa; Pupin, Breno; Dias, Luciana Pereira Pinto; Santos, Maiara P.; Bernardes, Luana Oliveira; Pinheiro, Michele Souza; Ferreira, Paulo Cesar; Mikaelyan, Aram; Medina, Humberto Ramírez; Alder-Rangel, AleneThe Mucor circinelloides complex, a group of dung-dwelling fungi, is essential for nutrient cycling and organic matter breakdown. Despite their ecological significance, variations in germination rates and stress tolerance among isolates are not well understood. This study examined four isolates—Cicarelli, Doc Beaver, Papulsa, and Zaina—to assess their germination on various media, including potato dextrose agar (PDA), Czapek (CZA), Sabouraud dextrose agar (SDA), Emerson (EM), and minimal media (MM). The results showed notable differ- ences in germination rates, with CZA and EM promoting the fastest growth. The study also analysed spor- angiospore tolerance to UV-B radiation and high temperatures (47 ◦C), with findings highlighting the role of growth media in stress resistance. Sporangiospores from Cicarelli, Doc Beaver, and Papulsa grown on PDA exhibited greater UV-B tolerance than those on CZA. Conversely, sporangiospores produced on CZA were more heat tolerant than those on PDA. The 12-h survival curve for the Zaina isolate reinforced these findings, showing similar trends in UV-B resilience and heat tolerance. This research demonstrates how growth media influence the environmental stress responses in M. circinelloides, providing insights into the adaptive potential of these fungi.