Doutorado em Biotecnologia RENORBIO

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A insegurança alimentar e nutricional : correlação epigenética do gene BDNF, status social e de saúde em agricultores familiares de café do Caparaó capixaba
(Universidade Federal do Espírito Santo, 2017-02-17) Barbosa, Wagner Miranda; Silva, Adriana Madeira Alvares da; Paula, Heberth de; Paula, Flávia de; Simão, João Batista Pavesi; Santos, Marcelo dos
Background: The association between insecurity about food and nutrition (INSAN) and violation of the human right to food (HRFN) is already known. In addition, the association between work on small family farms, poor health conditions, and INSAN has been proposed in several countries. However, the impact of living with INSAN on epigenetics, including the use of methylation studies has received little attention. The brain-derived neurotrophic factor, or BDNF, is important for the maintenance of brain functions, but stress can cause methylation of the promoter region of the BDNF gene, and this epigenetic change is associated with depression. Objective: To evaluate factors that might be associated with INSAN, including the methylation of exon I of BDNF and the living conditions of family farmers. Methods: Farmers from small family farms, 18 to 60 years of age, from 22 rural communities of 11 cities in the state of South Espirito Santo, southeastern Brazil participated in the study by answering a questionnaire about socioeconomic characteristics, ownership and use of land, behavior, health, and working conditions. The INSAN evaluation was carried out using the Brazilian Food Insecurity Scale. Methylation of exon I of the BDNF gene was examined by methylation-specific PCR. Body mass index (BMI) and biochemical and molecular analyses were performed. Logistic regression models were used to verify factors associated with INSAN. Results: The prevalence of INSAN and methylation were 23.68% and 39.01%. The occurrence of INSAN was associated the methylation of the exon I promoter of the BDNF gene, Odds Ratio (ORa) = 5.03 (95% CI 1.98 – 12.83); extra work off one’s own property, ORa = 3.36 (95% CI 1.23 – 9.21); land size (hectares), ORa = 0.77 (95% CI 0.67 - 0.90); and number of real-life symptoms and/or diseases, ORa = 1.12 (95% CI 1.04-1.20). Conclusions: The occurrence of INSAN in the members of small family farms, correlates with the conditions of a higher methylation rate of BDNF, the need to work outside the home because of low income, smaller size of the property, and greater numbers of diseases/symptoms reports.
Ação da nicotina na indução da expressão dos genes PIK3CA, AKT1, HIF-1α, GLUT1 e CA9 em linhagens celulares SCC9 e DOK
(Universidade Federal do Espírito Santo, 2021-02-24) Santos, Joaquim Gasparini dos; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108; https://orcid.org/0000-0001-6096-116X; http://lattes.cnpq.br/2629284844712364; Guimarães, Marco Cesar Cunegundes; https://orcid.org/0000000321460180; http://lattes.cnpq.br/0261991057482057; Santos, Marcelo dos; https://orcid.org/; http://lattes.cnpq.br/9871702190449067; Carvalho, Marcos Brasilino de; https://orcid.org/0000-0001-6854-2680; http://lattes.cnpq.br/6208433886573740
Nicotine is the main component present in tobacco products capable of activating pathways linked to tumorigenesis, tumor growth, metabolic changes and therapeutic resistance in several types of cancer, mainly in oral cavity cancer, but little is known about the effects of nicotine on lesions with malignant potential present in the oral cavity. In this work we seek to understand the effects of nicotine on the expression of genes linked to the pathway of tumor progression, such as PIK3CA, AKT1, HIF-1α, GLUT1 and CA9, in cell lines of DOK and SCC9. For that, the cell lines were submitted to cell viability assay by SRB, migration by wound test and analysis by RT-qPCR for expression of the genes of interest. Nicotine demonstrated the ability to increase the cell viability of SCC9 in all concentrations and in the time of 8 hours in relation to the control, the same was not observed in the wound test. In DOK cells, only the concentration of 10 µM, in the time of 8 hours, was able to increase the cellular migration in relation to the control. The ability of nicotine to increase gene expression in SCC9 was more evident at 24 hours and only the PIK3CA gene had a higher expression at 8 hours. In DOK it can be seen that the expression of the genes in question was restricted to the time of 8 hours and the concentration of 10 µM. Therefore, there is a relationship between exposure to nicotine and increased expression of genes in the SCC9 and DOK strains, to which the use of products with high concentrations of nicotine may contribute to the increased risk of malignization in regions with lesions with malignant potential present in the oral cavity.
Adaptação do usuário de próteses mioelétricas : implicações na aprendizagem dos movimentos da mão
(Universidade Federal do Espírito Santo, 2017-03-30) Costa, Regina Mamede; Bastos Filho, Teodiano Freire; Vargas, Cláudia Domingues; Zamora, Roberto Sagaró; Delis, Alberto López; Nogueira, Breno Valentim
From the physiological point of view, the knowledge of the anatomical and functional structure of the hand is essential for the understanding of the ostheomyoarticular mechanisms responsible for the movements of the fingers and their relation to the grasping functions. When injury occurs in one of these structures, the hand can be impaired, losing all functions as in the cases of amputation of the upper limb. The use of surface electromyography to control upper limb prostheses is an important clinical option, which offers the amputated an autonomy of control through the contraction of residual muscles. The functional biomechanical complexity of the hand involves a large area of representation in the cerebral cortex. In general, motor learning aims to maintain existing skills, the re-acquisition of lost skills and the learning of new skills. The goal of this Ph.D thesis was to propose and evaluate the application of a new experimental protocol for adaptation to the myoelectric prosthesis based on the distinction of hand movement patterns captured by sEMG of the remaining limb of amputees using myoelectric signals (SMEs).Ten upper limb amputees of both sexes, mean age 38.4 years ± 14.58, were evaluated. The inclusion criteria were: (1) transradial amputation or disarticulation of the wrist, unilateral or bilateral; (2) show no neurological or musculoskeletal disorder; (3) present no restriction of joint mobility. All of them were previously assessed including aspects of identification, anamnesis and physical examination. For MES acquisition, four Ag/AgCl active bipolar electrodes were used (TouchBionic ®). All the electrodes were placed according to the SENIAM (Surface Electromyography for the Non-Invasive Assessment of Muscle) recommendation. To the MES digitalization, an National Instrument NI USB-9001 acquisition system was used, and to the visualization of the MES, a digital processing software was developed, with interface to Matlab. The experimental protocol established a total of thirteen movements, which were grouped into two categories: GA (individual finger movements and hands opening and closure) and GB (grasping movements). The participants executed the tasks in three consecutive days. Two schemes were defined for the data capture: training phase and validation phase. The data concerning the first session (S1) were used to obtain a model of mechanical learning to the patterns classification, and the second (S2) and third (S3) sessions were used for the system validation. Although all tasks were performed in the same experiment, each category was studied and analyzed independently. Effectiveness (Acc), Kappa Coefficient (k) and Specificity (Sp) were calculated to evaluate the performance of each classifier of the executed movement. Positive-Negative Measurement (PNM) indicator was used to measure the performance of the thirteen proposed movements, and Goal Attainment Scale (GAS) was used to assess the extent to which individual objectives of each user were reached during the intervention. During the sessions, there were differences in the performance of the subjects during the proposed movements, which means that some participants could easily maintain repeated patterns, even with few training sessions, while others may need a longer training time to ensure good performance. Regarding the results of effectiveness, specificity, Kappa coefficient and PNM, the fact that the tasks of group A are simpler may explain the better performance of the volunteers in this group of tasks in relation to the performance in the tasks of group B (GB). On the other hand, the values obtained by GAS showed a satisfactory amount of correctness for the objectives outlined. Thus, this study showed that the subjects were able to perform muscular contractions, that is, perform the same movement with distinguishable MES patterns in the three experimental sessions, therefore, the proposed experimental design was validated in all the amputees of this study.
Alteração de expressão gênica em células mononucleares do sangue periférico humano submetidas à exposição com herbicida à base de glifosato
(Universidade Federal do Espírito Santo, 2018-06-27) Agostini, Lidiane Pignaton; Louro, Iúri Drumond; Paula, Flávia de; Guimarães, Marco Cesar Cunegundes; Vasallo, Dalton Valentim; Errera, Flávia Ambroisi Valle
Glyphosate is a post-emergent, non-selective and systemic herbicide. In the creating process of glyphosate-based herbicides (GBHs) such as Roundup®, surfactants are added to improve efficiency. The priority route of glyphosate’s degradation in soil results in aminomethylphosphonic acid (AMPA). Molecular responses to glyphosate were analyzed in some species of plants and in some vertebrates. In humans, it is not known exactly what risks and mechanisms of action would explain glyphosate’s toxicity reported in some experiments. The hypothesis is that fast exposure to Roundup® and AMPA leads to the differentiated expression of genes related to important cellular processes. Thus, the aim was identified these genes in human peripheral blood mononuclear cells when exposed to Roundup® and AMPA. The MTT [3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test was performed in triplicates to evaluate cell viability and the choice of treatment conditions used in the microarray technique (GeneChip® Human Transcriptome Array 2.0, Affymetrix). Eight chips were used: 3 for controls, 3 for AMPA (10 mM) and 2 for Roundup® (0.05%). The exposure time was 3 hours. Using a p <0.05 and a fold-change of ≥1.5 and ≤ −1.5, there were 26 differentially expressed genes (DEGs) identified after Roundup® exposure (3h; 0.05%) and 5 DEGs after AMPA treatment (3h; 10 mM). DEGs after Roundup® treatment showed association with 33 Gene Ontology (GO) cellular processes (enrichment analysis), mainly related to regulation. Pathview web was used to identify the effect off DEGs in different pathways. Only genes differentially expressed in Roundup® treatment were included in the pathways. TNF, LTA, TAB2 and ATM genes are related to NF-kappa B signaling pathway; BCL2L11 and ATM genes to FoxO signaling pathway; SESN3 and ATM genes to p53 signaling pathway; and TNF, BCL2L11 and ATM genes to apoptosis. Our results suggest that Roundup® change expression pattern of a several genes associated with cell cycle control, cellular processes regulation and apoptosis.
Análise da expressão de genes relacionados à adipogênese e à inflamação em tecido adiposo de mulheres com obesidade grau III
(Universidade Federal do Espírito Santo, 2013-04-29) Dasilio, Karine Lourenzone de Araujo; Errera, Flavia Imbroisi Valle; Paula, Flávia de; Louro, Iuri Drumond; Nogueira, Breno Valetim; Guiomarães, Marco Cesar Cunegundes; Costalonga, Everlayny Fiorot; Pereira, Thiago de Melo Costa
Obesity is a chronic condition defined as abnormal or excessive fat accumulation that impairs health. Overweight is an important risk factor for mortality and is a major risk factor for noncommunicable diseases, such as cardiovascular disease, impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM). Deposits of white adipose tissue (WAT), such as subcutaneous (SAT) and visceral (VAT), are biologically distinct, consequently, exhibited different biological effects on the health. WAT produces and secretes a variety of adipokines, such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), adiponectin (ADIPOQ), secreted frizzled-related protein 1 (SFRP1) and vascular endothelial growth factor (VEGFA), that are important in several cell functions, including angiogenesis, inflammation and adipogenesis. Alterations in adipokines expression might contribute to the onset of co-morbidities of obesity. The activation of wingless-type (WNT)/β-catenin pathway inhibits of adipogenesis via the blockade of CCAAT/enhancer binding protein alpha (CEBPA) and the peroxisome proliferator-activated receptor gamma (PPARG), which are essential transcriptional regulators of adipocyte differentiation. Evidence showed that non-fibrillar collagen XVIII (COL18A1), a protein of the extracellular matrix, has been shown to be active in adipogenesis. The aim of this study was, through quantitative Polymerase Chain Reaction, evaluate the expression of genes involved in adipogenesis and inflammation in SAT and VAT of women with severe obesity. The main results showed that i) the expression of the gene CEBPA in SAT is higher than in VAT of women with obesity ii) SAT CEBPA gene expression is higher in obese women with T2DM/IGT than in obese women with normal tolerance glucose tolerance (NGT), iii) the expression of SFRP1 gene in VAT was lower in women with T2D/IGT than those with NGT, iv) VAT COL18A1 gene expression was associated with better HDL levels and tolerance glucose. These results showed that the expression of genes involved in adipogenesis and inflammation differs in distinct WAT deposits and the expression of these genes were correlated with the metabolic parameters in obese individuals. This work contributes to a better understanding of the expression of genes related to these biological processes in SAT and VAT of patients with severe obesity.
Análise da expressão de genes relacionados à baixa densidade mineral óssea : avaliação prognóstica e de conduta terapêutica para osteoporose
(Universidade Federal do Espírito Santo, 2013-11-22) Souza, Leticia Soncini de; Bastos Filho, Teodiano Freire; Silva, Ian Victor; Nogueira, Breno Valentim; Guimarães, Marco César Cunegundes; Houzel, Jean Christophe; Pires, Rita Gomes Wanderley
Since that assuming the knowledge of the regulatory mechanisms of bone formation and resorption is crucial in the search for alternative therapies in diseases such as osteoporosis, molecular biology emerges as interesting tool and essential to achieving this goal. Among these genetic markers, gene polymorphisms associated with estrogen receptor alpha (ERα) and the gene Apolipoprotein " E" (ApoE) have received increased attention in recent years. The aim of this study was to analyze the expression of polymorphisms PvuII (CT nt -397) and XbaI (GA nt -351) gene present in REα and polymorphisms in the gene ApoE (HhaI - ε2 , ε3 and ε4) in populations of postmenopausal women, associating these gene alterations, and their clinical and biochemical profiles with osteoporosis. After analyzing the results obtained, the PvuII SNP in the gene of ERα is related to low BMD, and this effect is most noticeable in women with advanced age. The P allele, however, correlates strongly with high BMD (p < 0.05) in the whole population studied and reproduced when analyzing the population aged over 65 years, suggesting a protective role in the loss of bone mineral. In XbaII SNP, of same gene, a significant association of allele x at concentrations of triglycerides and total lipids and dependence on patient age and Body Mass Index (BMI) was observed. On the other hand, for SNP HhaI, ApoE gene, the E2 allele may be associated as a risk factor for low BMD, and E3 allele may be associated as a protective factor in relation to BMD. These results contribute to a better understanding of the expression of genes related to osteoporosis and can provide information for better prognostic determination of the disease as well as rationalization of therapeutic conduct to be chosen, providing a better quality of life for postmenopausal patients.
Análise elementar em carcinoma epidermoide de cavidade oral e sua relação com o tabagismo, expressão de genes de hipóxia e sobrevida
(Universidade Federal do Espírito Santo, 2018-09-25) Archanjo, Anderson Barros; Pinheiro, Christiano Jorge Gomes; Silva, Adriana Madeira Alvares da; Nogueira, Breno Valentim; Errera, Flavia Imbroisi Valle; Santos, Marcelo dos; Trivilin, Leonardo Oliveira
Head and neck cancer is a highly malignant and aggressive tumor type that presents a high rate of morbidity and mortality, mainly affecting the oral cavity, larynx and pharynx. The correlation between trace elements and the carcinogenesis process is due to the cytotoxic potential of some of these when available in excess concentrations. In addition to this, recent studies have described the association of these elements with the regulation of pathways related to tumor progression and the prognosis to be observed, among these pathways is the hypoxia pathway that has its main regulator HIF1. Thus, this work evaluated the elementary characterization in squamous cell carcinoma samples of the oral cavity, as well as the association of tobacco use in the elemental profile and the relation with the prognosis and survival of the patients. In addition, it evaluated the expression of the HIF1-α and PHD3 proteins and their relation to the smoking and alcohol habits, as well as the expression of these proteins to the presence of chemical elements in the same samples of squamous cell carcinoma of the oral cavity. For this, 78 samples of tumor tissue were obtained from patients diagnosed with squamous cell carcinoma of the oral cavity treated at the Arnaldo Vieira de Carvalho Cancer Institute and followed up for at least 24 months from initial treatment. The samples were categorized according to the smoking and alcohol habits. Tissue microarrays were made for elementary characterization using the synchrotron radiation-based μ-XRF technique, as well as for the analysis of the expression of HIF-1α and PHD3 proteins. The Chi-square test and the Fisher exact test were used for the association tests. Multivariate logistic regression by modeling was used to adjust odds ratios and confidence intervals. Survival curves were evaluated according to the Kaplan-Meier model and the Wilcoxon p-value and Cox Proportional Hazards model were used to adjust the hazards ratio values and the confidence interval. The following elements were detected: magnesium, phosphorus, sulfur, chlorine, potassium, calcium, chromium, manganese, iron, zinc, cobalt, nickel, copper, arsenic and bromine. Regarding the prognosis, magnesium was related to disease-free survival and chlorine and chromium with relapse, showing a protective factor against the occurrence of relapse and better disease-free survival. Nucleic HIF-1α expression was verified in 68 samples and cytoplasmic HIF-1α in 71 samples of oral cavity squamous cell carcinoma. Nuclear expression of PHD3 was observed in 49 samples and of cytoplasmic PHD3 in 48 samples. It was also verified the relation of the tumor size with the intensity of nuclear HIF-1α labeling and the positivity of the cytoplasmic PHD3 expression with the advanced clinical stage. It has also been shown that T3 / T4 tumors and the presence of chlorine increase the chances of presenting strong expression of cytoplasmic HIF-1α and that the clinical stage, the current consumption of alcohol and the presence of the chromium element influence the expression of PHD3. For survival, the association of disease-free survival with lymph node status was observed, while disease-specific survival was associated with lymph node status and advanced clinical stage. The present work was able to characterize oral squamous cell carcinoma samples for the presence of magnesium, phosphorus, sulfur, chlorine, potassium, calcium, chromium, manganese, iron, zinc, cobalt, nickel, copper, arsenic and bromine, of which the elements magnesium, chlorine, chromium, manganese, nickel, arsenic and bromine were associated with smoking. In addition, the chlorine and chromium elements were associated with the prognosis, in which the chlorine element was identified as an independent protective factor for recurrence and for better disease-free survival. Chromium, advanced clinical stage and alcohol consumption influenced the expression intensity of the PHD3 protein and the chlorine element and the tumor size of HIF-1α protein expression. This study presents unpublished data on the elemental profile, habits and expression of HIF-1α and PHD3 with the prognostic factors and survival of patients with squamous cell carcinoma of the oral cavity.
Análise funcional de nove SNPs de susceptibilidade ao câncer de ovário no locus 8q21
(Universidade Federal do Espírito Santo, 2018-03-19) Lyra Junior, Paulo Cilas Morais; Monteiro, Álvaro; Rangel, Letícia Batista Azevedo; Daltoé, Renata Dalmaschio; Gimba, Etel Rodrigues Pereira; Guimarães, Marco César Cunegundes; Barauna, Valério Garrone
Ovarian câncer (OC) is one of the most lethal cancer because of lack of early diagnosis. Genetic factors contributing to OC have recently been investigated through genome-wide association studies (GWAS), implicating in several independent OC risk loci in different chromosomal regions, among them the locus 8q21. Here, we performed an in-depth functional analysis of 9 SNPs causal candidates in the risk locus on 8q21 proximal to the CHMP4C gene. We first characterized the region for likely regulatory elements and genes associated, then we tested the 9 SNPs candidates for allele-specific activity in two different enhancer scanning assay, as well transcription factors associated.
Análise transcriptômica da interação de abacaxizeiro com o Pineapple mealybug wilt-associated virus
(Universidade Federal do Espírito Santo, 2018-06-29) Peron, Fernanda Nunes; Ventura, José Aires; Fernandes, Patricia Machado Bueno; Fernandes, Antonio Alberto Ribeiro; Carrer, Helaine; Rodrigues, Silas Pessini; Oliveira, Edna Maria Morais
Ananas comosus var comosus is a fruit of great economic and nutritional value worldwide. The productivity of the pineapple crop is influenced by mealybug wilt of pineapple (MWP). This disease is caused by Pineapple mealybug wilt-associated virus (PMWaV), the PMWaV-1 and PMWaV-2 variants are associated with the symptoms of the disease. The symptoms derive from the atrophy of the roots followed by wilting and discoloration of the leaves with redness and consequent deficiency in fruiting. The control of the disease occurs by the removal of symptomatic plants is not satisfactory especially since infected asymptomatic plants serve as a source of virus dispersion through their seedlings. Therefore, in this work, the differential transcriptome between symptomatic and asymptomatic infected plants was evaluated by RNA-seq, bioinformatic tools and RT-qPCR to propose an understanding of the pathogenesis of MWP under field conditions. Additionally, the proteins were confirmed by mass spectrometry analysis. Based on the reference genome of Ananas comosus, 16,097 expressed genes were identified, 268 repressed and 122 induced. The performance of RNA-seq was confirmed for 14 expressively expressed genes (DEGs) with a satisfactory Pearson correlation (R = 0.788). Functional classification and enrichment analysis revealed induction of genes involved in the regulation of flowering while repressed genes were predominantly related to the defense mechanisms of abiotic and biotic stress. Among them, some transcription factors (FT) WRKYs and MYBs, PRs, HSPs, AQPs and genes encoding ROS-removing enzymes, and Copper, Calcium and Zinc transporters were repressed. On the other hand, the expression of auxin responsive genes was positively regulated by ARFs. We observed hormonal regulation mediated by inhibition of jasmonate biosynthesis (JA), induction of ethylene biosynthesis (ET) and induction of the expression of genes responsive to auxin, what was related to the development of symptoms. A protein-protein interaction network was predicted allowing the visualization of the interaction of the gene products of the DEGs. Therefore, it was possible to observe the grouping of chaperones in the center of the network. We confirmed the inhibition of expression of the genes encoding the ERDJ3B, BiP2 and RTM2 chaperones in symptomatic plants and identified a significant negative correlation (R = -0.715) between the levels of RTM2 and PMWaV-2 transcripts. As the expression of this virus is predominant in symptomatic plants, we propose RTM2 as a probable gene associated with PMWaV-2 dispersion control. In addition, an HSP20 protein was identified only in asymptomatic plant samples reinforcing the hypothesis of HSP20 involvement in infection control. In the same way, we identified the HSP70, CaM and CRT proteins. These proteins had the repressed gene expression in the symptomatic plants pointing to stress in the endoplasmic reticulum (ER) as a response to PMWaV infection. Because viruses require the cellular machinery, we believe that pineapple plants suppress the expression of resident chaperones in the UPR (unfolfed protein response) as a strategy to limit the pathogen at the site of infection. However, with the introduction of SAR (systemic acquired resistance) and RTM2 expression, possible barriers to the displacement of PMWaV-2 were suppressed. In addition, the IRE1 expression reveals the activation of UPR to cell death by apoptosis. Thus, it is surprisingly concluded that PMWaV-2 infection elicits a hypersensitive response through UPR, in addition to modulating R gene expression.In view of the above, this thesis contributes to the revision of the methods adopted for the management of MWP both in cultural practices and the level of genetic improvement of pineapple.
APPLICATION OF GREEN NANOPARTICLES AND NANOCELLULOSE IN THE PRODUCTION OF SMART TEXTILES
(Universidade Federal do Espírito Santo, 2022-08-31) Silva, Laryssa Pinheiro Costa; Guimaraes, Marco Cesar Cunegundes; https://orcid.org/0000000321460180; http://lattes.cnpq.br/0261991057482057; https://orcid.org/; http://lattes.cnpq.br/; Dutra, Rosa Amalia Fireman; https://orcid.org/; http://lattes.cnpq.br/; Ventura, Jose Aires; https://orcid.org/0000000314221739; http://lattes.cnpq.br/8687116881326074; Nascimento, Diego Magalhaes do; https://orcid.org/; http://lattes.cnpq.br/
abstract
Aspectos clínicos e genéticos em uma população de idosos portadores de alterações cognitivas
(Universidade Federal do Espírito Santo, 2018-03-12) Magnago, Rafaella Papalino Lopes; Vieira, Fernando Luiz Herkenhoff; Silva, Ian Victor; Barauna, Valério Garrone; Guimarães, Marco Cesar Cunegundes; Campos, Luciene Cristina Gastalho
Aging is a global phenomenon already established in North America, Europe and in steady growth in countries in economic growth such as Brazil India and China. In Brazil, the changes occur in a radical and accelerated way. Alzheimer's disease is the most prevalent type of dementia, about 60% of dementias are Alzheimer's, the development of studies seeking to elucidate its pathogenesis has gained wide attention. The present study aimed to recognize epidemiological and biological data directly linked to dementia to prospect for updated forms of diagnosis not only by clinical signs that are more accessible and less invasive to the population. A search was made in the community of the great victory, of elderly people with or without cognitive loss. The population was divided into two groups, control (GC) and case (SHAZ), according to analysis of two predictive dementia tests (MMSE and CDR). From then on, clinical data were analyzed, searching for the prevalence of chronic diseases, the lipidogram of recent examinations of the individuals, a physical examination of predictive indexes of obesity as weight, percentage of fat, BMI, perimetry, of functional capacity, together with blood collection for DNA analysis and presence of polymorphisms of the genes ApoE, ERS1 for estrogen receptor α and PON1 (rs622 and rs705379). The results suggest that chronic diseases such as dyslipidemia and hypertension are risk factors in the development of dementia as well as age. The weight and the patterns of evaluation of obesity may have relation with the pathogenesis, however in this work, its interrelation is not clear. BMI can be considered substandard to correlate with disease. Functional capacity is dependent on the cognition factor and physical activity may delay the onset of the disease. The presence of genetic factors, linked to environmental factors exist and need to be further elucidated. The ApoE E3 and E4 alleles are alleles directly related to the disease and are suggested to be age dependent and biomarkers of vulnerability. The synergism between the ApoE E4 allele with ERS1 polymorphisms (PP, pp) and PON 1 (rs622-AA1 and rs 705379-AG2 and GG2) may be suggested as biomarkers, increasing between 3.2 and 5 times the odds of its development, coexisting multiple factors that have importance in the pathogenesis of this disease more and more prevalent in the elderly population worldwide.
Aspectos genéticos e terapêuticos da osteogênese imperfeita
(Universidade Federal do Espírito Santo, 2017-12-20) Moraes, Marcos Vinícius Dornelas de; Paula, Flavia de; Louro, Iúri Drumond; Guimarães, Marco Cesar Cunegundes; Batitucci, Maria do Carmo Pimentel; Lenz, Dominik
Osteogenesis imperfecta (OI) is a genetically heterogeneous hereditary disorder with an incidence of 1 in every 10-20,000 individuals born. It is characterized by deformities in connective tissue and bone fragility, which makes the individual with OI more susceptible to fractures, due to minimal trauma or non-traumatic impacts. In most cases the disease is inherited in an autosomal dominant (AD) form due to a mutation in heterozygosity in the COL1A1 or COL1A2 genes, which encode the collagen α1(I) and α2(I) chains of the collagen type I, one of the most important structural proteins of bones, skin and tendons. However, the number of reports of autosomal recessive mutations (RA) in new genes is increasing, and mutations in 14 genes directly related to the clinical expression of OI, including CRTAP, P3H1, PPIB, SERPINF1, SERPINH1, FKBP10, SP7 and WNT1, focus of this work. The use of bisphosphonates by intravenous administration has been the standard treatment in OI. The main benefits are the reduction of the number of fractures, increase of bone mass and reduction of chronic pain, which contributes to the control of the progression of the disease a significant improvement in the patients' quality of life.
Associação do papaya meleira virus e de um segundo vírus de ssRNA à meleira do mamoeiro
(Universidade Federal do Espírito Santo, 2017-01-23) Antunes, Tathiana Ferreira Sá; Fernandes, Patricia Machado Bueno; Fernandes, Antônio Alberto Ribeiro; Silva, Diolina Moura; Ventura, José Aires; Silva, Maite Vaslin de Freitas
Papaya sticky disease, or “meleira”, is one of the major diseases of papaya in Brazil and Mexico, capable of causing complete crop loss. The causal agent of sticky disease was identified as an isometric virus with a double stranded RNA (dsRNA) genome, named papaya meleira virus (PMeV). In the present study, the nearly complete sequence was obtained for PMeV dsRNA which 8,814 nucleotides long and contains two putative ORFs; the predicted ORF1 and ORF2 display similarity to capsid proteins and RdRp's, respectively, from mycoviruses tentatively classified in the family Totiviridae. Besides PMeV dsRNA, a second RNA band of approximately 4.5 kb was isolated from latex of papaya plants with severe symptoms of sticky disease. The sequence obtained for the second RNA is 4,515 nucleotides long and contains two putative ORFs. The predicted ORFs 1 and 2 display 48% and 73% sequence identity, respectively, with the corresponding proteins of papaya virus Q, an umbravirus recently described infecting papaya in Ecuador. Viral purification in a sucrose gradient allowed separation of particles containing each RNA. Mass spectrometry analysis indicated that both PMeV and the second RNA (named papaya meleira virus 2, PMeV2) were encapsidated in particles formed by the protein encoded by PMeV ORF1. The presence of both PMeV and PMeV2 was confirmed in field plants showing typical symptoms of sticky disease. Interestingly, PMeV was detected alone in asymptomatic plants. Together, our results indicate that sticky disease is associated with double infection by PMeV and PMeV2.
ATORVASTATINA AUMENTA A EFICÁCIA DA CISPLATINA EM CÉLULAS DE CÂNCER DE MAMA ATRAVÉS DA MODULAÇÃO DO METABOLISMO LIPÍDICO
(Universidade Federal do Espírito Santo, 2021-11-12) Santos, Diandra Zipinotti dos; Rangel, Leticia Batista Azevedo; https://orcid.org/0000000261504064; http://lattes.cnpq.br/6706127386696883; https://orcid.org/0000000204565303; http://lattes.cnpq.br/1926925060779955; Monteiro, Robson de Queiroz; http://orcid.org/0000-0002-5260-2332; http://lattes.cnpq.br/0848413770343284; Guimarães, Marco Cesar Cunegundes; https://orcid.org/0000000321460180; http://lattes.cnpq.br/0261991057482057; Stringari, Lorenzzo Lyrio; https://orcid.org/0000-0002-8710-3648; http://lattes.cnpq.br/8291250597662563; Nimrichter, Leonardo; https://orcid.org/0000-0001-9281-6856; http://lattes.cnpq.br/1080378998643497
Acquired treatment resistance is a major problem in breast cancer (BC) management, especially aggressive subtypes such as triple-negative-BC (TNBC). Understanding tumor biology and designing new treatments is critical to improving therapeutic efficacy and prognosis. In some cancers, alterations in lipid metabolism, including the accumulation of cholesterol and cholesteryl esters (CE), contribute to tumor progression and development of drug resistance. The present study aimed identify the role of cholesterol metabolism in BC response to cisplatin (CDDP) treatment in the acute setting and in acquired resistance, and if atorvastatin (ATV) can modulate platinum sensitivity. In vitro studies in MCF-7 (Luminal A), MDA-MB-231 (TNBC) and in a developed CDDP-resistant MDA-MB-231 cells (MDACR) were performed using a variety of biochemical and radiometric techniques. Cell viability and cell proliferation were measured through MTT assay and Incucyte, respectively. We demonstrated that CDDP alters cholesterol metabolism in luminal A (MCF -7) and TNBC (MDA-MB-231) cell models and that CDDP in combination with low concentrations of ATV reduces cell proliferation and cell viability more than CDDP alone, especially in MDA-MB-231 cells. MDA-MB-231 cells display elevated levels of CE, LDLR and ACAT1 levels compared to MCF-7 cells, whereas MCF-7 cells had greater HMGCR protein levels compared to MDA-MB-231 cells. The combination of CDDP and ATV dramatically altered cell cholesterol metabolism more in MDA-MB-231 cells than in MCF -7 cells. These results suggest that the increased susceptibility of MDA-MB-231 cells to co-administration of CDDP and ATV compared to MCF -7 cells is related to an increased dependence on CE availability. ATV restored CDDP sensitivity in CDDP-resistant cells, MDACR, which exhibit upregulation of ACAT-1 compared to their parental cells MDA-MB-231. Overall, our data suggest that association of ATV with conventional antineoplastic drugs appears to be beneficial, especially when associated with CDDP for the MDA-MB-231 lineage. Moreover, our data from cell culture-based studies supports upregulation of cholesterol homeostasis as an adaptive response that contributes to aggressiveness and chemotherapy resistance. This adaptive behavior of breast cancer cells and targeting of cholesterol metabolism warrants further preclinical and clinical investigation as a novel strategy to overcome CDDP resistance.
Aumento da atividade celulásica para hidrólise da casca de coco verde pela aplicação da alta pressão hidrostática
(Universidade Federal do Espírito Santo, 2013-04-01) Albuquerque, Érica Dutra; Fernandes, Antonio Alberto Ribeiro; Fernandes, Patricia Machado Bueno; Marco, Janice Lisboa De; Torres, Fernando Araripe Gonçalves; Castro, Eustáqio Vinícius; Gomes, Daniel Cláudio de Oliveira
The use of cellulosic biomass for cellulosic ethanol production has been a serious option for the production of renewable fuels. The main focus in the cellulosic ethanol production is to increase the hydrolysis of these cellulosic materials in order to produce products with a low-molecular weight, such as hexoses.The high costs involved with this process due to the use of cellulases and the biomass pretreatment, causes the main problem in an economically viable large-scale production of bioethanol. Even though enzymatic disruption of cellulose has been considered the best environmental practice, cellulase is the most expensive step during cellulosic ethanol production. Another problem is the inhibition of the cellulase complex by reducing-sugars such as cellobiose and glucose. Significant cost-reduction becomes needed in order to enhance viable commercial use of cellulase hydrolysis technology. Using coconut husk as a biomass substrate, we applied two approaches: firstly, we isolated fungi from coconut husks. We found that their cellulases outperforme the cellulases of the industry standards on the hydrolysis of coconut husk. Secondly, we employed High Hydrostatic Pressure (HHP) to the cellulases in order to improve the enzymatic performance. After the HHP treatment, we observed an increase in coconut husk hydrolysis by a factor of 2 as a result of an increase of the βglucosidase, endoglucanase and total cellulase activities. The effect of HHP is able to break the hydrogen bonds working down to the scales of the coconut fibers. Further, the inhibition of β-glucosidases caused by glucose was removed under HHP, the βglucosidases achivied higher activities in the presence of 55 mM of glucose as a result of changes in the conformation of this cellulase on HHP. Using bioinformatics tools to analyse structures of β-glucosidase and endoglucanase, we found hydrophobic aminoacids and glycosylation regions in the cavities; in the active sites; and on β-sheets structures. While HHP affects the cavities of the proteins, the carbohydrates can bind both to their side chains of hydrophobic amino acids as to glycosylation regions, leading to an increase of the activities and a stabilization of the protein. HHP to value of 300 MPa exposes the internal apolar parts of the coconut fiber (CEF) and favors the binding of hydrophobic aminoacids of the cellulase to these parts of the fibers, as well as to glucose. The increase of the cellulase hydrolysis under HHP can be caused by a factor that works like a CBM (Carbohydrate Binding Module) that approaches the substrate at the active site. Thus, employing both the 21 technologies HHP as well as substrate specific fungi can potentially make bioethanol production an economic viable and energy-efficient process, especially regarding the improvement of β-glucosidase activities.
Avaliação de marcadores moleculares relacionados ao prognóstico de pacientes com carcinoma epidermóide de cavidade oral e orofaringe
(Universidade Federal do Espírito Santo, 2014-08-29) Santos, Marcelo dos; Conforti, Adriana Madeira Álvares da Silva; Louro, Iuri Drumond; Paula, Flávia de; Nogueira, Breno Valentim; Guimarães, Marco Cesar Cunegundes; Tajara, Eloiza Helena
Head and neck cancer is the fourth in incidence and the fifth in mortality among the most frequent malignancies worldwide. For 2014, there has been estimated over 15 thousand new oral and oropharynx cancers in Brasil. Similar to other tumors, oral and oropharynx cancer is a multifactorial disease, caused by multiple environmental and genetic factors, involving alterations in molecular pathways and cellular homeostasis. The investigation of molecular markers involved in this process has been the object of my studies, especially because in spite of great advances in the molecular aspects of cancer, little progress has been made in the clinical outcome during the last decades. It is known that tumor progression depends on cellular aquisition of competences, such as apoptosis evasion, cell proliferation disregulation, angiogenic activation, cellular adaptation to hipoxia, cell survival mechanism activation and hypoxia dependent epigenetic changes. Therefore, the present work had the purpose to evaluate the potential of proteins FAS, FASL, FGFR4, LEPR, HIF1-a, NDRG1 and JMJD1a, as well as polymorphisms FGFR4 Gly388Arg and LEPR Gln223Arg, as putative molecular markers for clinicopathological tumor features or oral and oropharynx squamous cell carcinoma. Our results show that HIF1-1 expression was associated with local disease relapse and local disease-free survival in patients who undertook pos-operative radiotherapy and were also related to tumor vascular microdensity. In addition, NDRG1 expression was different in tumor tissue and non tumoral margins, but also showing an association with disease survival. FGFR4 polymorphism and expression showed a relation with death and disease survival. However, FGFR4 expression alone showed an association with lymph node metastasis and relapse. FGFR4 profile showed a relation with disease survival. FAS/FASL expression showed a correlation with death and its proposed profise was related with disease survival. LEPR Gln223Arg polymorphism was realted with disease free survival and disease specific survival, whereas LEPR expression was realted with lymph node metastasis. JMJD1A nuclear and cytoplasmic expression were related with lymph node metastasis. However, only nuclear expression was related with relapse and disease survival. In conclusion, our results suggest that proteins ans polymorphisms can be used as molecular markers to help predict prognosis in oral and oropharynx squamous cell carcinoma.
Avaliação de usabilidade, desempenho ocupacional e satisfação com sistema de controle de ambiente inteligente por pessoas com deficiência motora severa através de eletromiografia de superfície e oculografia por infravermelho
(Universidade Federal do Espírito Santo, 2019-06-13) Sime, Mariana Midori; Bastos Filho, Teodiano Freire; Silva, Adriana Madeira Álvares da; Frizera Neto, Anselmo; Cruz, Daniel Marinho Cezar da; Naves, Eduardo Lázaro Martins
A smart environment (SE) is an Assistive Technology (AT) resource that allows people with motor disabilities, even with low mobility, to control the lighting and electronic equipment (TV, radio, fan, etc) of the environment through a Human-Machine Interface (HMI) configured to be activated by biomedical signals. However, despite the recognized importance, the AT resource is not always considered useful, reaching high abandonment rates, since a prior assessment and prescription by professionals is necessary, taking into account the real demands and needs of the person with disability. This Doctoral Thesis aims to evaluate the effectiveness of a smart environment system controlled by surface electromyography and by infrared oculography, captured by an eye tracker, used by people with motor disabilities. Six volunteers participated in the research, and, initially, were applied socio-demographic data forms, Functional Independence Measure (FIM TM) and Canadian Occupational Performance Measure (COPM). The subjects were presented to the equipment and system interface, being trained for their use in domestic environment, using the system for a week. Afterwards, they were re-evaluated with the COPM, besides evaluations of satisfaction with the use of the AT resource (form B-QUEST 2.0), psychosocial impact (form PIADS), usability of the system (SUS form) and semi-structured interview for suggestions or complaints. The control of TV was the common demand of all participants. As a result of this research, of the six volunteers, four used the system, presenting positive results regarding the change in occupational performance, satisfaction with performance and the smart environment system, high psychosocial impact and good system usability. It was evaluated that the developed system also provided greater independence of the volunteers for the control of the equipment. Regarding the volunteers who did not use the system, aspects such as nonacceptance of the disability and lack of social support may have influenced. The SE system proved to be effective, improving all aspects evaluated in the participants. A patent application of the developed system was submitted to INIT-UFES. Future studies should consider expanding the possibilities of controlled equipment and devices, as well as the time of use.
Avaliação do senso de presença em ambiente de realidade virtual para reabilitação motora de pacientes pós-AVE : uma abordagem baseada em EEG
(Universidade Federal do Espírito Santo, 2019-02-18) Pomer-Escher, Alexandre Geraldo; Bastos Filho, Teodiano Freire; Rodriguez, Denis Delisle; Bravo, Eduardo Caicedo; Nogueira, Breno Valentim; Fernandes, Antonio Alberto Ribeiro
abstract
AVALIAÇÃO DO SENSO DE PRESENÇA EM AMBIENTE DE REALIDADE VIRTUAL PARA REABILITAÇÃO MOTORA DE PACIENTES PÓS-AVE: UMA ABORDAGEM BASEADA EM EEG
(Universidade Federal do Espírito Santo, 2019-02-18) Escher, Alexandre Geraldo Pomer; Filho, Teodiano Freire Bastos; https://orcid.org/0000000211852773; http://lattes.cnpq.br/3761585497791105; https://orcid.org/; http://lattes.cnpq.br/; Fernandes, Antonio Alberto Ribeiro; https://orcid.org/; http://lattes.cnpq.br/4696507759154477; Nogueira, Breno Valentim; https://orcid.org/0000000221990635; http://lattes.cnpq.br/0011229320439147; Rodriguez, Denis Delisle; https://orcid.org/000000028937031X; http://lattes.cnpq.br/7140331839822423; Bravo, Eduardo Francisco Caicedo
abstract
Avaliação e caracterização de insumos bioativos da aroeira (Schinus terebinthifolius Raddi) com potencial econômico para o desenvolvimento tecnológico de bioprodutos
(Universidade Federal do Espírito Santo, 2013-06-20) Sales, Maria Diana Cerqueira; Fernandes, Patricia Machado Bueno; Ventura, José Aires; Costa, Hélcio; Kuster, Ricardo Machado; Meira, Débora Dummer; França, Hildegardo Seibert
Aroeira (Schinus terebinthifolius Raddi) is an important plant mainly due its industrial application in the food industry, cosmetics and pharmaceutics. Since society has been exigent to choice qualified products, in this work we aimed identification and evaluation of bioactive products, such as mother tinctures (MT), extracts (Eb) and essential oils (EO) from leaves and fruits of aroeira genotypes. The TM were obtained from the leaves of plants hermaphrodite (TMArFH), female (TMArFF), and male (TMArFM) and fruits from hermaphrodite plants (TMArFrH) and two samples of female plants (and TMArFrF1 TMArFrF2), were prepared from the plant extract (with natural adjuvant) by maceration. OE from leaves (F01-F04) and fruits (the FR01- FR07), were obtained from the hydrodistillation. Analytical methods were used to perform physic-chemical analyses of MT and chemistry compounds identification were carried out by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), gas chromatography (GC) and gas chromatography-Mass spectrometry (GC/MS). The antioxidant activity of the different MT was evaluated using the stable free radical DPPH (1, 1- diphenyl-2- picryl-hydrazyl) method and, evaluation of antifungal efficiency of TM was performed by the agar diffusion method, for the fungi Fusarium guttiforme and Chalara paradoxa, pineapple pathogens. The physico-chemical analysis showed average values of dry residue (1.41 ± 0.13%), alcohol content (67% ± 0.78), pH (5.6 ± 0.78) and density (0.898 g. mL-1 ± 0.98), within the technical specifications, indicating quality in the process and specifications to obtaining the TM. The chromatograms showed the presence of phenolic groups, especially, isolation of two important biflavonoids, agathisflavona and tetrahidroamentoflavona. Monoterpenes predominated, and the sample FR01, had the highest percentage of limonene (36.85%), which represented the highest relative content (70.85%). Regarding the antioxidant activity (AA), the TMArFH (250 µg/mL), showed better availability for uptake of DPPH (AA= 59.09%). In the antifungal test both TMArFH as tebuconazole fungicide at 0.1%, showed strong inhibition of the mycelial growth for F. guttiforme (PI= 82%) and a significant efficiency for C. paradoxa (PI= 42. 86%). This is the first report with the use of MT from leaves and fruits of S. terebinthifolius with potential feedstock for the development of bioproducts with fungicidal activity for control plant pathogens.

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