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Influência mitocondrial na regulação de genes envolvidos com envelhecimento em Saccharomyces cerevisiae submetidas à alta pressão hidrostática
(Universidade Federal do Espírito Santo, 2020-02-07) Spagnol, Brigida de Almeida Amorim; Fernandes, Patricia Machado Bueno; https://orcid.org/0000000326953638; http://lattes.cnpq.br/2441925791593067; https://orcid.org/0000-0001-5482-2702; http://lattes.cnpq.br/3200148222923666; Lomeli, Monica Montero; https://orcid.org/0000-0003-0449-7563; http://lattes.cnpq.br/9488252687145315; Fernandes, Antonio Alberto Ribeiro; https://orcid.org/0000-0003-0535-9349; http://lattes.cnpq.br/4696507759154477; Santos, Alexandre Martins Costa; https://orcid.org/0000000288018875; http://lattes.cnpq.br/4144105396879016; Pinto, Nadja Cristhina de Souza; https://orcid.org/0000-0003-4206-964X; http://lattes.cnpq.br/7088639503480810
Saccharomyces cerevisiae is a well-accepted eukaryotic model organism for studies on stress response and aging, whereas preserves several biochemical, molecular and organelle functioning mechanisms. Mitochondria is an organelle involved with stress response and, because it is main generator of reactive oxygen species (ROS) is strongly implicated with aging. Dysfunctionalities of this organelle has been associated with neurodegenerative, psychiatric, and cancer diseases. Mature and young S. cerevisiae cells respond differently to high hydrostatic pressure (HHP) stress. Cell maturity favors genes regulation involved with aging, increasing by almost three times the HHP stress resistance of these cells compared to young cells. Aging regulatory genes such as TOR1, RAS2 and SCH9 have been implicated with mitochondrial dysfunctions. However, it is possible that mitochondria influence the aging-related genes regulation. Thus, this work sought to modulate the TOR1, RAS2 and SCH9 regulation by controlling the increase in mitochondrial membrane potential (ΔΨm) of S. cerevisiae submitted to HHP. To achieve these goals, S. cerevisiae mother cells were differentiated from daughter cells by Calcofluor White impregnationinto their cell wall, generating daughter cells without impregnation. Thus, ROS accumulation and ΔΨm in response to HHP was quantified by flow cytometry. Protocatechuic acid (PCA) was previously used for mitochondrial protection against the HHP unfavorable effects. Fluorescence microscopy images on mitotracker green- FM labeled cells were performed to observe mitochondrial dynamics. Finally, genes involved with aging and antioxidant defenses were analyzed by real-time PCR (qRT- PCR) in the general population. Mother and daughter cells indicatived similarities in ΔΨm upon the applied treatments. Mild HHP stress indicated ΔΨm increased and 15 min recovery at ambient pressure are required for ΔΨm stabilization before severe HHP treatment. Mitochondrial protection indicated reducing of ΔΨm and this seems to have improved the tolerance to severe treatment. Mature cells accumulated high levels of ROS after HHP. However, mitochondrial protection together with the PCA antioxidant action reduced the ROS accumulation in these cells, compensating the inhibition of antioxidant genes regulation. Almost all cells with mitochondrial protection resisted to severe HHP stress. There was a 71% to 92% increase in HHP stress tolerance with previous mitochondrial protection. The genes TOR1, RAS2 and SCH9 were repressed, favoring the stress response, increasing the resistance. In addition, reducing ROS accumulation has retarded aging.
Fluorescência da clorofila a como indicador de porta enxertos de manga ‘Ubá’ mais tolerantes ao déficit hídrico
(Universidade Federal do Espírito Santo, 2020-02-13) Silva, Leonardo Faria; Silva, Diolina Moura; https://orcid.org/000000033885280X; http://lattes.cnpq.br/0341541450627705; https://orcid.org/0000000192822818; http://lattes.cnpq.br/; Santos, Alexandre Martins Costa; https://orcid.org/0000000288018875; http://lattes.cnpq.br/4144105396879016; Araujo, Wagner Luiz; https://orcid.org/; http://lattes.cnpq.br/; Ventura, Jose Aires; https://orcid.org/0000000314221739; http://lattes.cnpq.br/8687116881326074; Nesi, Adriano Nunes
In order to select the photochemical parameters extracted from the chlorophyll a fluorescence kinetics as being the simplest, fastest and most efficient for early identification of the most vigorous rootstock in the formation of drought tolerant 'Ubá' see
INFECÇÃO POR PAPILOMAVÍRUS HUMANO E EXPRESSÃO DAS PROTEÍNAS P16 E EGFR COMO BIOMARCADORES DE PROGNÓSTICO EM CARCINOMA EPIDERMOIDE ORAL E OROFARÍNGEO
(Universidade Federal do Espírito Santo, 2020-03-30) Abreu, Priscila Marinho de; Zeidler, Sandra Lucia Ventorin Von; https://orcid.org/0000000288975747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000000164537171; http://lattes.cnpq.br/; Rotta, Rejane Faria Ribeiro; https://orcid.org/; http://lattes.cnpq.br/; Rabenhorst, Silvia Helena Barem; https://orcid.org/; http://lattes.cnpq.br/; Paneto, Greiciane Gaburro; https://orcid.org/0000000180354199; http://lattes.cnpq.br/8176374147579841; Paula, Flavia de; https://orcid.org/0000000186792982; http://lattes.cnpq.br/7913201450663683; Podesta, Jose Roberto Vasconcelos de
Oral and oropharyngeal squamous cell carcinoma has as main risk factors the consumption of tobacco, alcohol and infection by Human Papillomavirus (HPV). Studies indicate that HPV-positive patients have results more favorable in relation to survival and re
Sazonalidade, epidemiologia molecular e virulência do Vírus Sincicial Respiratório (VSR): uma perspectiva dentro do programa brasileiro de vigilância da influenza
(Universidade Federal do Espírito Santo, 2021-05-24) Vianna, Lucas Alves; Louro, Iuri Drumond; https://orcid.org/0000000151609615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0000-0001-8487-9929; http://lattes.cnpq.br/6931449640908148; Arruda Neto, Eurico de; https://orcid.org/0000-0002-0978-410X; http://lattes.cnpq.br/1632065399563444; Fernandes, Patricia Machado Bueno; https://orcid.org/0000000326953638; http://lattes.cnpq.br/2441925791593067; Spano, Liliana Cruz; https://orcid.org/0000000262056988; http://lattes.cnpq.br/7451382455806895; Silva, Paola Cristina Resende; https://orcid.org/0000-0002-2884-3662; http://lattes.cnpq.br/3725179069720464; Sousa, Rita Catarina Medeiros; https://orcid.org/0000-0002-8330-6667; http://lattes.cnpq.br/3560941703812539
Respiratory Syncytial Virus (RSV) is the main cause of pediatric morbidity and mortality. The complex evolution of RSV creates a need for worldwide surveillance, which may assist in the understanding of multiple viral aspects. This study aimed to investigate RSV features under the Brazilian Influenza Surveillance Program, evaluating the role of viral load and genetic diversity in disease severity and the influence of climatic factors in viral seasonality. We have investigated the prevalence of RSV in children up to 3 years old with severe acute respiratory infection (SARI) in the Espirito Santo State (ES), Brazil, from 2016 to 2018. RT-qPCR allowed for viral detection and viral load quantification, to evaluate association with clinical features and mapping of local viral seasonality. Gene G sequencing and phylogenetic reconstruction demonstrated local genetic diversity. Of 632 evaluated cases, 56% were caused by RSV, with both subtypes A and B co-circulating throughout the years. A discrete inverse association between average temperature and viral circulation was observed. No correlation between viral load and disease severity was observed, but children infected with RSV-A presented higher clinical severity score (CSS) median, stayed longer in the hospital, required intensive care and ventilatory support more frequently than those infected by RSV-B. Regarding RSV diversity, some local genetic groups were observed in the main genotypes circulation RSV-A ON1 and RSV-B BA, with strains showing modifications in the G gene amino acid chain. Local RSV studies using the Brazilian Influenza Surveillance Program are relevant to assess the suitability and viability of a single network for the surveillance of other respiratory viruses. Understanding seasonality, virulence and genetic diversity can support the suitability of future antiviral drugs and vaccines and assist in the administration of prophylactic strategies.
Análise da excitabilidade e conectividade cortical para um sistema de neurorreabilitação baseado em interface-cérebro máquina e monociclo robótico
(Universidade Federal do Espírito Santo, 2021-06-29) Cardoso, Vivianne Flávia; Bastos Filho, Teodiano Freire; https://orcid.org/0000000211852773; http://lattes.cnpq.br/3761585497791105; https://orcid.org/0000-0002-1654-3320; http://lattes.cnpq.br/1122896995621553; Nogueira, Breno Valentim; https://orcid.org/0000000221990635; http://lattes.cnpq.br/0011229320439147; Andrade, Adriano de Oliveira; https://orcid.org/0000-0002-5689-6606; http://lattes.cnpq.br/1229329519982110; Caldeira, Eliete Maria de Oliveira; https://orcid.org/0000-0002-3742-0952; http://lattes.cnpq.br/8508979665054143; Frizera Neto, Anselmo; https://orcid.org/0000000206873967; http://lattes.cnpq.br/8928890008799265; Rodriguez, Denis Delisle; https://orcid.org/000000028937031X; http://lattes.cnpq.br/7140331839822423
Recently, studies on cycling-based brain-computer interfaces (BCIs) have been standing out due to their potential for lower-limb recovery. In this scenario, the behavior of the sensory motor rhythms and the brain connectivity present themselves as source of information that can contribute to interpret the cortical effect of these technologies. This study aims to analyze how motor sensory rhythms and the cortical connectivity behave when volunteers command a reactive motor imagery (MI) BCI that provides passive pedaling feedback. Eight healthy subjects performed pedaling MI to command an Electroencephalography (EEG) based BCI with motorized pedal to receive passive movements as feedback. The EEG data were analyzed under the following four conditions: resting, MI calibration, MI online and receiving passive pedaling (online phase). Most subjects produced around Cz, corresponding to the foot area significant event-related desynchronization (ERD) patterns around Cz, when performing MI and receiving passive pedaling. The sharpest decrease was found for low β band (13 – 22 Hz). The connectivity results revealed an exchange of information between supplementary motor area (SMA) and parietal regions during MI and passive pedaling. Findings point the primary motor cortex activation for most participants and the connectivity between SMA and parietal regions during pedaling MI and passive pedaling.
Fatores determinantes da dor crônica e o papel da metilação do gene NR3C1
(Universidade Federal do Espírito Santo, 2021-08-30) Branco, Alexandre Lima Castelo; Louro, Iuri Drumond; https://orcid.org/0000000151609615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0000-0002-1704-9877; http://lattes.cnpq.br/7332472547330240; Guimarães, Marco Cesar Cunegundes; https://orcid.org/0000000321460180; http://lattes.cnpq.br/0261991057482057; Arantes, Lidia Maria Rebolho Batista; https://orcid.org/0000-0001-8230-1218; http://lattes.cnpq.br/2019308149950531; Carvalho, Marcos Brasilino de; https://orcid.org/0000-0001-6854-2680; http://lattes.cnpq.br/6208433886573740; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108
Chronic pain is a multidimensional health condition with high prevalence in Brazil and its chronic condition may be related to depression and anxiety, diseases recognized as the most prevalent mental disorders in the world and major causes of functional incapacity, suffering and reduced quality of life. The relationship between depression, anxiety, pain, suffering and epigenetic alterations have already been described in the literature, but this relationship is not completely clear yet. Epigenetic alterations can affect gene expression and are related to the individual's adaptation to the environment in a relationship between genotype, phenotype and environment. The glucocorticoid receptor gene, NR3C1, is regulated by epigenetic mechanisms and acts to control the neuroendocrine axis via cortisol, which also links the gene to depression and other psychiatric illnesses. Thus, this research evaluated the determinants of chronic pain, biopsychosocial, biochemical and molecular factors in the epigenetic modifications of the NR3C1 gene in adults aged between 20 and 59 years, users of the Brazilian Unified Health System. The results of the biopsychosocial assessment in the sample showed a profile of people over 40 years old, with lower per capita income and education, low levels of cortisol, more reports of stress and anxiety, higher consumption of continuous medications, less physical activity and more prevalence of chronic pain. Pain was related to a statistical model that pointed out pain-related variables. Thus, the research showed indicators that point to a predominant profile of individuals with chronic pain, with determinant factors being: hypomethylation of the DNA of the NR3C1 gene in CpG 42, age over 40 years and low cortisol.
Variabilidade genética em NOTCH1: associação com o excesso de peso e implicações terapêuticas na Leucemia Linfocítica Crônica em uma coorte de idosos
(Universidade Federal do Espírito Santo, 2021-09-03) Barcelos, Estevão Carlos Silva; Zeidler, Sandra Lúcia Ventorin Von; https://orcid.org/0000000288975747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000-0001-7076-6165; http://lattes.cnpq.br/6899125943013073; Maranduba, Carlos Magno da Costa; https://orcid.org/0000-0001-7327-1934; http://lattes.cnpq.br/4763153859701731; Naslavsky, Michel Satya; https://orcid.org/0000-0002-9068-1713; http://lattes.cnpq.br/5084104783599564; Nogueira, Breno Valentim; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; Paula, Flavia de; https://orcid.org/0000000186792982; http://lattes.cnpq.br/7913201450663683
NOTCH1 is critical for the maintenance of stem cells and adult tissues, and appears to play as a key molecule in aging and in a variety of chronic conditions, including obesity and cancer, especially Chronic Lymphocytic Leukemia (CLL). Recently, the NOTCH1 signaling pathway has been identified as fundamental in adipocyte homeostasis, in addition to being directly linked to the inflammatory process. Furthermore, constitutive activation of NOTCH1 signaling is associated with poor prognosis in some types of cancer such as CLL where recurrent NOTCH1 mutations are responsible for stabilizing the signaling state. Given the involvement of the NOTCH1 gene in cell metabolism and development, the hypothesis of the present study is that genetic variations of NOTCH1 are associated with increased risk for overweight (overweight and obesity) and also alter the response of CLL cells to induced-stress. Therefore, as a first objective, we investigated whether genetic variations in NOTCH1 are associated with overweight/obesity in a Brazilian elderly cohort. To assess the association of NOTCH1 polymorphisms and overweight/obesity, anthropometric, biochemical, medical history, and lifestyle data were collected. Sequencing data from 1.024 individuals (aged 59 to 99 years) were analyzed: 424 were overweight, 320 were obese, and 280 were normal weight. We analyzed one hundred and sixty-one tag SNPs spanning the entire NOTCH1 gene and borders using the SNP tagging approach (minor allele frequency≥0.01 and pairwise linkage disequilibrium r2≥0.8). We observed the association of SNP rs9411207 with risk of overweight/obesity under the additive model, and the genotype distribution showed an increased frequency of homozygous TT (OR = 1.50, 95% CI: 1.20-1.88; P = 0.0002). The GAT haplotype constructed from this and other SNPs in high linkage disequilibrium (LD) was more frequent in overweight/obese subjects (P = 0.003). In silico analysis suggested that these SNPs likely affect the transcription of NOTCH1 and other genes. This is the first study reporting the association between NOTCH1 SNPs and risk of overweight/obesity. Considering the possibility of modulation of NOTCH1, additional population studies are needed to replicate these results and confirm the usefulness of these risk genotypes for new therapeutic strategies. A second objective was to evaluate whether the mutation in the NOTCH1 gene and the deregulation of the NOTCH1 pathway in CLL cases alter the response to curcumin. For this, we investigated the anti-CLL effect of curcumin and its ability to interfere in the integrated stress response (ISR) and in the NOTCH1 signaling pathway in primary CLL cells with mutation in the NOTCH1 gene and in the murine model of CLL Eμ-TCL1. Initially, gene expression data from cells with c.7544- 7545 delCT mutation in NOTCH1 were analyzed against data from Wild-Type (WT) cells. In silico analysis of gene expression of CLL cells mutated in NOTCH1 showed an increase in the expression of genes involved in Endoplasmic Reticulum (ER) and ISR stress. Primary CLL cells were collected and cultured with curcumin, a natural compound that also targets ER stress, for further analysis by Western blotting and real-time PCR. After in vitro exposure to curcumin, stress-related mechanisms were observed in primary cells with mutated NOTCH1 compared to WT cells. This response was preceded by an early increase in Ca2+ in the cytoplasm of CLL cells mutated in NOTCH1, which can trigger and maintain ER stress. Furthermore, curcumin increased the apoptosis of CLL cells carrying the NOTCH1 mutation, regardless of allelic load, when compared to WT cells. NOTCH1 signaling also appears modulated after exposure to curcumin. In particular, CLL cells mutated in NOTCH1 showed a reduction in the active portion of NOTCH1 (ICN1) and in the antiapoptotic proteins of the BCL2 family (BCL2 and MCL1), and the combination of curcumin with venetoclax, an anti-BCL2, shown to have a synergistic effect on CLL cells. In addition, we used the murine model Eμ-TCL1 to evaluate treatment with curcumin. In vivo administration of curcumin in the Eμ-TCL1 model significantly reduced the percentage of CD5+/CD19+ leukemic cells infiltrating the liver, spleen and bone marrow, with concomitant inhibition of NOTCH1 signaling in leukemic cells in the bone marrow. Our results suggest that ER stress-induced ISR activation and NOTCH1 signaling inhibition converge to amplification of CLL cell death and provide a crucial target for CLL treatment.
Novas abordagens no diagnóstico não invasivo de câncer bucal utilizando espectroscopia ftir em amostras de sangue e saliva
(Universidade Federal do Espírito Santo, 2021-09-29) Thebit, Marcela Marçal; Gouvea, Sonia Alves; https://orcid.org/000000015180471X; http://lattes.cnpq.br/7268228122543743; https://orcid.org/0000-0002-7305-5753; http://lattes.cnpq.br/7117284804597883; Abreu, Gláucia Rodrigues de; https://orcid.org/0009-0008-8772-8470; http://lattes.cnpq.br/0229590907405570; Endlich, Patrick Wander; https://orcid.org/0000-0002-7597-3603; http://lattes.cnpq.br/9669721267985878; Carvalho, Luis Felipe das Chagas e Silva de; https://orcid.org/0000-0003-1063-4624; http://lattes.cnpq.br/5669726117269568; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108
Oral cancer is usually diagnosed through invasive methods, for although systemic changes can be easily detected, the identification of an effective systemic biomarker using a simple and affordable approach has yet to take place. Attenuated total reflection Fourier transform infrared (ATR-FTIR) appears as an alternative for the chemical and structural characterization of organic and inorganic materials, including liquid samples as serum, plasma and saliva and a possible bridge between molecular analysis and clinical practice. Objective: The aim of this study was to evaluate if ATR-FTIR spectroscopy can be applied as a tool in oral cancer screening through the analysis of plasma and salivary samples. Methodology: Plasma and saliva samples from oral cancer patients and controls were analyzed by ATR-FTIR spectroscopy. Experimental data were fed into the MATLAB 2018b software and them analyzed through two different routines in order to identify points of differentiations between the groups. Results: Biochemical analysis of the plasma samples through ATR-FTIR yielded 92% sensitivity and 100% specificity in differentiating between groups. The biggest difference was found between peaks that might be related to nucleic acids and nucleic acid phosphate. Pilot study with salivary samples reached similar accuracy Conclusion: We conclude that ATR-FTIR successfully discriminated between oral cancer patients and healthy subjects using plasma samples, with nucleic acids being found as major biomarkers.
Dynamics of the papaya meleira virus complex during the development of papaya (Carica papaya L.)
(Universidade Federal do Espírito Santo, 2021-11-11) Araujo, Marlonni Maurastoni; Fernandes, Patricia Machado Bueno; https://orcid.org/0000000326953638; http://lattes.cnpq.br/2441925791593067; https://orcid.org/0000-0002-6064-3126; http://lattes.cnpq.br/6052184072658200; Aragão, Francisco José Lima; https://orcid.org/0000-0003-0570-7673; http://lattes.cnpq.br/6121745125946782; Fernandes, Antonio Alberto Ribeiro; https://orcid.org/0000-0003-0535-9349; http://lattes.cnpq.br/4696507759154477; Zerbini Junior, Francisco Murilo; https://orcid.org/0000-0001-8617-0200; http://lattes.cnpq.br/6507826653693228; Rodrigues, Silas Pessini; https://orcid.org/0000-0002-8672-3626; http://lattes.cnpq.br/9067991933812274; Whitfield, Anna Elizabeth; https://orcid.org/0000-0002-3538-015X
Among the most serious virus-incited diseases in papaya production is papaya sticky disease (PSD). This disease was first reported in Brazil in 1993, associated with a double-stranded RNA virus, called papaya meleira virus (PMeV). Since then, progress has been made in the knowledge of the disease dispersion in the field, the etiological agent characterization, and its interactions with papaya. However, in 2016, the papaya meleira virus 2 (PMeV2), with a positive single-stranded RNA genome, was also identified in diseased plants, imposing a rethinking of the pathosystem. Therefore, in this work, we critically evaluate the latest findings on PSD and the last 30 years of research done to understand its dispersion in the field. We show that leafhopper and whitefly species need to be better studied as potential vectors of the PSD-associated viruses in Brazil now that more sensitive molecular diagnostic techniques are available. Nevertheless, we developed a multiplex RT-PCR (mPCR) technique capable of detecting both viruses in a single reaction from pre-flowering plant samples, which is a useful tool for the early diagnosis of PSD. Here we show that laticifers of the main vein of papaya sticky diseased leaves are the preferential infection site of PMeV and PMeV2. We also show that the PMeV capsid is composed of two major polypeptides with overlapping sequences. A central fragment of these polypeptides (aa 321-670) interacts with the 50S ribosomal protein L17 (RPL17), which we speculate as an important player in virus accumulation. Overall, this thesis discusses PSD in three main spheres: the biology of the etiological agent and its interaction with the host, the spread of the disease in the field, and the development of technologies for its management.
Valor prognóstico do infiltrado linfocitário tumoral e da expressão de PLK1 e FOXM1 em carcinoma epidermoide oral
(Universidade Federal do Espírito Santo, 2022-07-25) Damasceno, Thabata Coeli Dias; Zeidler, Sandra Lucia Ventorin Von; https://orcid.org/0000000288975747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000000244434025; http://lattes.cnpq.br/0520491056597962; Mendonça, Elismauro Francisco de; https://orcid.org/0000-0002-6751-3279; http://lattes.cnpq.br/2305019128015847; Lima, Sheila Coelho Soares; https://orcid.org/0000-0002-6742-3708; http://lattes.cnpq.br/7690812849140276; Peterle, Gabriela Tonini; https://orcid.org/0000-0001-7735-936X; http://lattes.cnpq.br/6804646896381238; Silva, Adriana Madeira Alvares da; https://orcid.org/0000000280780304; http://lattes.cnpq.br/6445492335035108
Oral squamous cell carcinoma (OSCC) has an estimated 377,713 new cases each year in the 2020-2022 triennium worldwide. Individuals affected by OSCC exhibit heterogeneous clinical behavior and diagnosis at an advanced stage, causing a worse prognosis. Early detection of the disease increases survival rates by up to 80%. Thus, the analysis of the dynamics between the immune response, through the analysis of the tumor lymphocytic infiltrate (TIL), and the tumor progression, evaluated by the use of the biomarkers PLK1 and FOXM1, can be essential tools to assist in the detection, recurrence and prognosis. of the disease. Serum PLK1 promoter region methylation, PLK1 and FOXM1 gene expression in tumor tissue, and Plk1 expression in tumor adjacent epithelium, dysplasia and tumor were evaluated as prognostic indicators in patients with OSCC. Plk1 expression was evaluated in 109 paraffinized tissue samples, 21 frozen tumor fragments and 30 serum (17 before treatment initiation and 13 after treatment). The Chi-Square and Fisher's Exact tests were used to establish an association between the variables studied with TIL density, methylation status and Plk1 expression; to compare the mean expression of Plk1 in the segments studied, the Mann-Whitney U test and the Bonferroni post-test were used; One-Way analysis of variance (ANOVA) and application of Dunnett and/or Tukey tests were applied to verify patterns of gene expression; and survival curves were evaluated using the Kaplan-Meier model. Low TIL density was associated with T3/T4 tumor size (p = 0.001) and clinical stage III/IV (p = 0.011); while high TIL was associated with T1/T2 tumor size (p = 0.001), clinical stage I/II (p = 0.01) and surgery as treatment performed (p = 0.046). The PLK1 methylation status showed no association with the variables analyzed. High expression of PLK1 was observed in 11 samples in relation to the control (p < 0.0001), as well as two samples had a different expression when in parity to the others (p < 0.0001). FOXM1 did not show expression in the sample group. The analysis of Plk1 expression in the segments showed a correlation between the adjacent epithelium pairs and the dysplasia and tumor regions (p < 0.001). High Plk1 expression was associated (p < 0.001) with the variables T3/4 tumor size, lymph node metastasis and stage III/IV. Likewise, low expression was associated (p < 0.001) with smaller tumor size, no lymph node involvement and early stages. Tumor Plk1 expression and TIL density were not shown to be prognostic factors when analyzed for the overall survival and disease free survival curves, but they did show to be related to tumor development and progression in OSCC.
Identificação de cochonilhas em cafeeiros utilizando DNA Barcoding e High Resolution Melting e distribuição potencial de Planococcus citri no Espírito Santo
(Universidade Federal do Espírito Santo, 2023-01-26) Oliveira, Pablo Viana; Paneto, Greiciane Gaburro; https://orcid.org/0000000180354199; http://lattes.cnpq.br/8176374147579841; https://orcid.org/000000021785178X; http://lattes.cnpq.br/9623188128443119; Serrão, José Eduardo; https://orcid.org/0000-0002-0477-4252; http://lattes.cnpq.br/6663553463256293; Telles, Mariana Pires de Campos; https://orcid.org/0000-0002-9023-0007; http://lattes.cnpq.br/4648436798023532; Ferreira, Marcia Flores da Silva; https://orcid.org/0000-0003-1541-6634; http://lattes.cnpq.br/5719813884063445; Ventura, José Aires; https://orcid.org/0000000314221739; http://lattes.cnpq.br/8687116881326074
Mealybugs (Hemiptera: Pseudococcidae) are agricultural pests of several crops, which can transmit viruses and cause great economic losses. For the control and management of these pests is essential a rapid and accurate identification of these insects. In this study, a rapid and low-cost method for identifying mealybug species was developed. The objectives are presented in the Results and Discussion section written in three chapters in Scientific Paper format. The first chapter presents an article accepted for publication in the Diversity journal, entitled "Molecular Species Delimitation Using COI Barcodes of Mealybugs (Hemiptera: Pseudococcidae) from Coffee Plants in Espírito Santo, Brazil". The dataset containing mitochondrial COI gene sequences of 26 putative of species Pseudococcidae recorded in Brazil were explored using the ASAP, GMYC, mPTP delimitation methods. An incongruence between the methods was observed, with the number of species ranging from 22 to 30 putative species. Of these, 10 species were identified among the new specimens collected in Brazil. In the second chapter, a paper to be submitted in a Journal with Impact Factor (≥ 3) is presented, entitled "Development of a High-Resolution Melting method based on COI minibarcodes to identify mealybug (Hemiptera: Pseudococcidae) pest species". A 76 bp primer pair was designed based on COI gene sequence alignment for 5 species of mealybugs. Analysis was performed by PCR immediately followed by HRM and the 5 species tested were discriminated, including two closely related species, Planococcus citri and Pl. minor. In addition, intraspecific variation was also detected in Pl. citri. The third chapter presents a paper for submission to a Journal with an Impact Factor (≥ 3), entitled "Potential distribution of the citrus mealybug, Planococcus citri (Risso, 1813), an important pest from coffee tree in Brazil”. The most abundant species found in coffee trees in Espirito Santo was chosen as a model for potential distribution prediction. Species distribution modelling was performed based on MaxEnt. North and northwest of the state was revealed to have the highest potential occurrence of this pest. This assumption was confirmed with our collection data, presented in the first chapter. This thesis presented important results on the diversity of species of mealybugs occurring in ES and another brazilian states. The tools used will help in the rapid identification of these insects and can be implemented in strategies for prophylaxis, monitoring and control of quarantine pests of agricultural crops in Brazil and other countries.
Activity of prokaryotic communities from offshore oil reservoir and their adaptation to high hydrostatic pressure
(Universidade Federal do Espírito Santo, 2023-02-15) Santos, Luiza Favarato; Fernandes, Patricia Machado Bueno; https://orcid.org/0000000326953638; http://lattes.cnpq.br/2441925791593067; https://orcid.org/0000000173409358; http://lattes.cnpq.br/1217579607633423; Fernandes, Antonio Alberto Ribeiro; https://orcid.org/0000-0003-0535-9349; http://lattes.cnpq.br/4696507759154477; Santos, Alexandre Martins Costa; https://orcid.org/0000000288018875; http://lattes.cnpq.br/4144105396879016; Rodrigues, Silas Pessini; https://orcid.org/0000-0002-8672-3626; http://lattes.cnpq.br/9067991933812274; Paula, Flavia de; https://orcid.org/0000000186792982; http://lattes.cnpq.br/7913201450663683; Biasi, Ronaldo Sergio de
Deep-sea microbes comprise a significant portion of Earth’s biomass, but they are still poorly studied. Although the subseafloor is an oligotrophic, anoxic, dark, and pressurized environment, life still thrives in such kind of environment. In this work, three samples collected 100 m, 3,060 m, and 6,000 m deep in the Brazilian coast were investigated by analysis of 16S rRNA. A difference was found in the biological diversity between these communities at the genus level. The three communities had acid-producing bacteria and sulfate-reducing bacteria, but only the community from 3,060 m deep had a methanogenic archaeon. From the 6,000 m deep sample was isolated a bacterium of the Halanaerobium genus. Two important variables for the environment and offshore industries, bacterial survival, and hydrogen sulfide (H2S) production under high hydrostatic pressure (HHP) were investigated. Cell survival and H2S production were affected by HHP. In the case of the 3,060 m bacteria, population and H2S production increase with increasing pressure, while for the 100 m bacteria, population is stable with increasing pressure but H2S production decreases. The results show the importance of studying the influence of pressure on bacterial growth and H2S production to define strategies to mitigate economic losses arising from activities in marine environments.
Análise da composição química e da atividade antibacteriana e antifúngica de clones de Schinus terebinthifolia Raddi
(Universidade Federal do Espírito Santo, 2023-03-01) Moraes, Rodrigo; Ventura, Jose Aires; https://orcid.org/0000-0003-1422-1739 ; http://lattes.cnpq.br/8687116881326074 ; https://orcid.org/0000-0001-9175-4825; http://lattes.cnpq.br/0598172220327707; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/4144105396879016; Endringer, Denise Coutinho; https://orcid.org/0000-0001-9396-2097; http://lattes.cnpq.br/6424432214565576; Kuster, Ricardo Machado; https://orcid.org/0000-0002-8961-5348; http://lattes.cnpq.br/4149814906786366; França, Hildegardo Seibert ; https://orcid.org/0000-0001-6129-8793; http://lattes.cnpq.br/1284874997224988
Aroeira (Schinus terebinthifolia Raddi) is included in the Brazilian List of Essential Medicines due to its potential to generate bioactive compounds, including those with antimicrobial properties. The occurrence of antimicrobial resistance increases health care costs, length of stay in hospitals, morbidity, and mortality. WHO has published a list of antibiotic-resistant “priority pathogens” to guide and promote research, development, and innovation of new antibiotics including those that may combat oxacillin-resistant Staphylococcus aureus and carbapenemase-producing Klebsiella pneumoniae. Fungi of the genus Fusarium are important pathogens for plants, humans, and animals, in addition to producing mycotoxins in food, with a great lack of new molecules that can be used for their control. This research sought to characterize the phytochemical profile of polar and non-polar extracts of mature fruits of four Aroeira clones and their antimicrobial activity against oxaxilin-resistant Staphylococcus aureus, carbapenemase-producing Klebsiella pneumoniae and the fungus Fusarium guttiforme. The extracts of all aroeira clones showed antimicrobial activity for bacteria and fungus, varying according to the clone and solvent used. Polar extracts showed greater antifungal and antibacterial activity, being more active on Gram-positive bacteria. The antifungal and antibacterial activity varied according to the evaluated clone. Clone BAAr 5 showed greater antibacterial activity while clones BAAr 5 and BAAr 8 had greater antifungal activity. The Aroeira clones showed similarity in the chromatographic profile by HPLC. It was possible to observe a greater relative number of peaks in clone BAAr 5 when extracted using a hydroethanolic solvent. The chromatogram of the hexane extracts showed a greater diversity of peaks. There was a greater abundance of sesquiterpenes compared to monoterpenes when the samples were evaluated in GC-MS. Spectrometry using ESI(-)FT-ICR MS identified the presence of 29 ions, between m/z 195 and m/z 925 in the polar extracts of the clones. Twenty-four ions were observed in clone BAAr 5, 20 in clone BAAr 8, 23 in clone BAAr9 and 17 in clone BAAr 23. Although the clones were grown under the same edaphoclimatic conditions, they showed variation in antimicrobial activity and bioactive compounds identified. It is suggested that the commercial use of Aroeira should come from selected and cloned plants that present the desired characteristics
Diversidade e frequência haplotípica de X-STRS na população do Espírito Santo e sua contribuição para elucidação de casos forenses complexos
(Universidade Federal do Espírito Santo, 2023-03-03) Rodrigues, Fernanda Mariano Garcia de Souza; Louro, Iuri Drumond; https://orcid.org/0000000151609615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0000000270355005; http://lattes.cnpq.br/0479360701927218; Meira, Debora Dummer; https://orcid.org/0000-0002-6092-2459; http://lattes.cnpq.br/7199119599752978; Carvalho, Elizeu Fagundes de; https://orcid.org/0000-0003-4620-7253; http://lattes.cnpq.br/2742420738858309; Paula, Flavia de; https://orcid.org/0000000186792982; http://lattes.cnpq.br/7913201450663683; Rave, Cintia Fridman
Genetic markers Short Tandem Repeats (STR) are the center of human genetic identification, STR markers on autosomal chromosomes and on the Y chromosome being the most used. However, in some forensic situations, such as cases of suspected incest, paternity without a maternal sample for comparison, traces with mixed DNA, the use of only these markers may not be enough to solve these cases. The study of STRs markers of the X sex chromosome (X-STRs) significantly increases the probability of identification by complementing the data obtained for autosomal and Y chromosome markers. Statistical analyzes that must be included in the issuance of expert reports. Thus, the general objective of this work is to carry out a macro-regional survey of the haplotype frequencies of 12 loci of X-STRs in Espírito Santo, to estimate the genetic diversity, the dynamics of populations and promote the gain of statistical power and the updating of this information in databases international data. In this sense, the survey of haplotype frequencies of X-STRs was carried out with a sample group of 571 unrelated individuals born in the 4 macro-regions of the state, in order to obtain a reliable estimate of the genetic diversity of ES. Analyzing the set of 12 XSTRs, no statistically significant differences were found between the macro regions of the state. The allele and haplotype frequencies found here have high rates of allelic and haplotype variability, demonstrating that this X-STR set is very informative in terms of individual discrimination, and may contribute to building knowledge and the importance of using the X chromosome in routine of laboratories that use DNA technology in human identification.
Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
(Universidade Federal do Espírito Santo, 2023-11-22) Có, Anna Clara Gregório; Camillo, Cláudia Malheiros Coutinho; https://orcid.org/0000-0001-9016-2668; http://lattes.cnpq.br/3184503163639480; Zeidler, Sandra Lúcia Ventorin von; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000-0002-7737-0371; http://lattes.cnpq.br/3678557620411441; Nunes, Fabio Daumas; https://orcid.org/0000-0002-7785-6785; http://lattes.cnpq.br/4909755821591847; Errera, Flavia Imbroisi Valle; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Arantes, Lidia Maria Rebolho Batista; https://orcid.org/0000-0001-8230-1218; http://lattes.cnpq.br/2019308149950531; Paula, Flavia de; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683
Oral squamous cell carcinoma (OSCC) is the 16th most commonly diagnosed form of cancer globally, with a higher prevalence in the tongue compared to other areas of the oral cavity. However, the lack of effective biomarkers for diagnosis, especially for precancerous lesions, poses a limitation, as visual or histological examination cannot predict the progression of dysplastic lesions, making it difficult to determine whether they will develop into cancer or return to normal epithelium. In this context, the present research aims to investigate molecular targets that may indicate the irreversible transformation of these cells, to provide a basis for broader studies aimed at using these targets as biomarkers for early OSCC diagnosis. To achieve this goal, this experimental study addressed the evaluation of the expression of a panel of microRNAs and proteins in tumour tissues and adjacent tumour-adjacent epithelium obtained from patients with tongue squamous cell carcinoma and oral epithelium from healthy individuals. Additionally, the research explored the association between these biomarkers, seeking to determine their potential application as diagnostic biomarkers for tongue squamous cell carcinoma. A total of 75 cases of tongue squamous cell carcinoma and adjacent tumouradjacent epithelium were included in the study, and the expression of the proteins survivin, Bcl-2, PLK1, p16, p40, p63, EGFR, and cyclin D1 was analysed by immunohistochemistry. The analysis of the microRNA panel's expression involved 31 samples of tongue squamous cell carcinoma, 10 samples of healthy gingival tissue, and 10 samples of serum from healthy individuals, as well as 7 samples of serum from patients diagnosed with OSCC, using the RT-qPCR technique. In silico analysis by bioinformatics validated the findings related to the expression of differentially expressed microRNAs in the sample group. The results showed differences in the expression of miRNA-31-5p (p<0.001) and miRNA-21-5p (p=0.001) in tumour samples compared to control samples. Significant differences were not observed in the expression of miRNA-24-3p, while miRNA-542-3p and 196a-5p were not detected in the sample group. No significant difference was observed in the expression of miRNAs in serum samples. The assessment of the diagnostic potential of microRNAs included ROC curve analysis, which revealed that miR-21-5p had an area under the curve (AUC) of 0.803, while miR-31-5p obtained an AUC of 0.777. The results also identified differential expression among the proteins survivin, PLK1, and p63, all of which showed increased expression in tumour tissue. Additionally, a correlation was observed between the expression of miR-21-5p and the protein p40 (chi-square: p=0.047; Spearman correlation: r=0.402; p=0.023). In conclusion, the results suggest that miR-21-5p and miR-31-5p may be potential diagnostic biomarkers for tongue squamous cell carcinoma, providing a foundation for further exploration for large-scale studies to explore miRNA-protein correlations, considering the site specificity of miRNAs.
Potencial prognóstico de células inflamatórias e PD-L1 solúvel em carcinoma epidermoide de cabeça e pescoço
(Universidade Federal do Espírito Santo, 2024-04-16) Daniel, Camila Batista; Zeidler, Sandra Lúcia Ventorin von ; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000-0003-3992-5986; http://lattes.cnpq.br/5905656000130424; Santos, Eldamária de Vargas Wolfgramm dos ; https://orcid.org/0000-0003-3815-0760; http://lattes.cnpq.br/4688343262832362; Errera, Flavia Imbroisi Valle ; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; Arantes, Lidia Maria Rebolho Batista ; https://orcid.org/0000-0001-8230-1218; http://lattes.cnpq.br/2019308149950531; Mendonça, Elismauro Francisco de ; https://orcid.org/0000-0002-6751-3279; https://orcid.org/0000-0002-6751-3279; http://lattes.cnpq.br/2305019128015847
Immunosuppression is recognized as a hallmark of cancer and has been associated with worse outcomes. In head and neck squamous cell carcinoma (HNSCC), immunosuppression is a hallmark of the tumour and may be mediated by immunosuppressive inflammatory cells and PD-L1 expression. Given their participation in this process, our study aimed to describe the prognostic impact of these elements in tumour tissue and peripheral blood HNSCC patients. We conducted a multicentre retrospective study with biological samples and clinical data from HNSCC patients recruited at two centres in Brazil and the United Kingdom and healthy individuals. To analyse the inflammatory infiltrate, we used tumour tissue slides stained with haematoxylin and eosin and immunohistochemistry for PD-L1 analysis and neutrophil quantification. Absolute leukocyte counts were retrieved from pretreatment blood counts available in medical records. To evaluate the prognostic value of PD-L1 in liquid biopsy, we used serum and plasma samples obtained from patients and healthy individuals for quantification of soluble PD-L1 (sPD-L1) by ELISA. Fresh tumour tissue samples were used to analyse CD274 gene expression levels using the RT-qPCR technique. Furthermore, we analysed PD-L1 expression by flow cytometry in HNSCC cell lines and quantified sPD-L1 levels in the supernatant. The tumour microenvironment analysis showed that low levels of lymphocytes are associated with worse overall survival and disease-free survival. However, we did not observe this relationship with tumour-associated neutrophils and tumour PD-L1. When analysed together in a scoring system, we demonstrated that low levels of lymphocytes, high expression of PD-L1 and high infiltration of neutrophils are associated with a worse outcome. In blood, a high ratio of neutrophils and lymphocytes (NLR) was also associated with worse survival but was not correlated with inflammatory components of the tumour. High levels of sPD-L1 protein were associated with reduced overall survival, however our study did not identify a relationship between sPD-L1 levels in the blood and PD-L1 expression in tumour tissue, determined by immunohistochemistry and RT-qPCR. In contrast, the in vitro study showed that the levels of sPD-L1 released 14 into the supernatant are strongly correlated with cytoplasmic and membrane expression. Our data further showed that sPD-L1 levels in patients were positively correlated with the number of leukocytes, neutrophils and monocytes in peripheral blood. Taken together, our results highlight the prognostic potential of markers of the tumour microenvironment analysed in a combined manner, in a scoring system, as a way of providing a more comprehensive overview of tumour behaviour, highlighting important events, such as immunosuppression. Our results highlight the prognostic potential of PD-L1 detected by liquid biopsy in HNSCC and indicate that the levels detected in the blood do not necessarily correspond to what is observed in the tumour. Therefore, it is believed that it should be analysed as an independent marker whose immunosuppressive role is carried out at a systemic level
Avaliação do potencial bioativo de frações peptídicas de coprodutos de peroá (balistes capriscus)
(Universidade Federal do Espírito Santo, 2024-04-30) Ribeiro, Monique Lopes; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Cicilini, Maria Aparecida; https://orcid.org/; http://lattes.cnpq.br/; Oliveira, Jairo Pinto de; https://orcid.org/; http://lattes.cnpq.br/; Koblitz, Maria Gabriela Bello; https://orcid.org/; http://lattes.cnpq.br/; Souza, Bartolomeu Warlene Silva de; http://lattes.cnpq.br/
Fish industry generates a significant amount of waste which have high biological value and potential for industrial use. The aim of this study was to evaluate the antioxidant and ACE-inhibitory activities of peptide fractions from triggerfish (Balistes capriscus) processing coproducts. Molecular mass distribution of the proteins extracted from fish viscera was determined by molecular exclusion chromatography presenting fractions with a wide range of molecular mass (<1.2 kDa to > 440 kDa). Soluble Protein Extract (SPE) was hydrolysate using papain (HP), bromelain (HB) and trypsin (HT) (3% p.p-1, 6h). Samples were fractionated (>100 MWCO, 30–100, 10-30 and < 10 MWCO). Antioxidant activity of fractions was evaluated and fraction SPE4 (<10 MWCO) showed the highest value of Trolox Equivalent Antioxidant Capacity - TEAC (10,157.7 μmol Trolox. g-1) and Ferric Reducing Antioxidant Power - FRAP (1,588.71 μmol FeSO4. g-1 ). SPE and hydrolysates (<10MWCO) were separated by ion-exchange chromatography. Fraction F1 showed the highest value for TEAC capacity (8,839.04 μmol Trolox. g-1) and FRAP (1,749.94 μmol FeSO4. g-1). ACE-inhibitory activity was evaluated for non-hydrolysate and hydrolysate fractions. Fractions F3, F5 and HP3 showed the lowest IC50 value (30.1, 42.7 e 37.7 µg, respectively). Antimicrobial activity was observed in samples SPE (against S. aureus), F1 and F4 (against Pseudomonas sp.) It was identified 20 amino acid sequences that could contribute to the biological activity of the peptide fractions. Proteins extracted from triggerfish viscera demonstrated to be a good source of bioactive peptides that may have food and pharmaceutical applications.
Transmissão por insetos do complexo papaya meleira virus (PMeV e PMeV2) e prospecção e caracterização de uma fração da proteína estrutural do PMeV
(Universidade Federal do Espírito Santo, 2024-05-06) Almeida, Joellington Marinho de; Santos, Alexandre Martins Costa ; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/4144105396879016; Fernandes, Patricia Machado Bueno ; https://orcid.org/0000-0003-2695-3638; http://lattes.cnpq.br/2441925791593067; https://orcid.org/0000-0003-4747-1398; http://lattes.cnpq.br/3138032439066350; Ventura, José Aires ; https://orcid.org/0000-0003-1422-1739; http://lattes.cnpq.br/8687116881326074; Xavier, André da Silva ; https://orcid.org/0000-0002-9251-0301; http://lattes.cnpq.br/5661020509713522; Araújo, Marlonni Maurastoni ; https://orcid.org/0000-0002-6064-3126; http://lattes.cnpq.br/6052184072658200; Rodrigues, Silas Pessini ; https://orcid.org/0000-0002-8672-3626; http://lattes.cnpq.br/9067991933812274
Papaya blight is a disease caused by a virus that can lead to a loss of productivity in Carica papaya orchards in producing countries such as Brazil, Mexico, Ecuador and Australia where the disease occurs. Infection of the plant causes burning of the tips of young leaves, spontaneous exudation of latex and spots on the fruit. The latex oxidizes in the presence of air, giving rise to a "honey-like" appearance, which together with other factors interferes with the commercial acceptance of the fruit. However, the vector that transmits the papaya late blight disease is still unknown in all producing countries. Studies carried out by various research groups have pointed to the leafhopper as a potential disseminator of the disease, in addition to the relationship with fungi, seeds and mechanical transmission. In Brazil, it is caused by the viral complex papaya meleira virus (PMeV) and papaya meleira virus 2 (PMeV2), and the coexistence of the two viruses has proven that PMeV2 is encapsidated by PMeV, and it is known that PMeV has two ORFs responsible for encoding structural proteins that make up the capsid (ORF1) and a putative RdRp protein (ORF2). Knowing that the structural proteins of PMeV are responsible for encapsidating the two genetic materials, the use of the expression of a recombinant protein fraction of the capsid protein (CP) of PMeV could become viable in the development of rapid tests, identification of viral particles in different locations of the plant and potential application in studies of virus-plant-vector transmission. In this study, we developed material on understanding the viral transmission of the papaya meleira virus complex by insects using the available literature and the prospecting and characterization of a fraction (p441) of the PMeV structural protein. The p441 fraction of the PMeV capsid protein was expressed in E. coli BL21(DE3) and extracted from the SDS-PAGE gel. The protein fraction was expressed at four different times (30 minutes, 1 hour, 2 hours and 4 hours) to check which would correspond to its highest production and its extracted fraction was used to produce polyclonal antibodies. Based on the available sequence, an in silico characterization was carried out to determine ab initio secondary and tertiary structure models, physicochemical parameters and the prediction of immunogenic peptides that could be identified in its primary sequence. The results obtained open up possibilities for the design of tests that are effective in detecting PMeV, based on observations made about the protein structure of the p441 fraction as being a stable protein with low thermal mobility. In addition, 18 peptides found on its surface have been shown to be capable of triggering an immune response and to be a region that can be used to produce antibodies that can be applied in future studies to locate the virus in different parts of the plant. Finally, it was found that the conditions of expression in a time of 4 hours obtained the best response, generating protein concentrations of around 633 to 923 mg/mL verified after SDS-PAGE gel extraction
Detecção de beauvericina por HPLC-DAD e MALDI-FT ICR MS em frutos de abacaxi infectados por Fusarium guttiforme
(Universidade Federal do Espírito Santo, 2024-06-27) Fávero, Carolinne Simões; Romão, Wanderson ; https://orcid.org/0000-0002-2254-6683; http://lattes.cnpq.br/9121022613112821; Ventura, José Aires ; https://orcid.org/0000-0003-1422-1739; http://lattes.cnpq.br/8687116881326074; https://orcid.org/0000-0002-0915-0350; http://lattes.cnpq.br/6385055031220477; Errera, Flavia Imbroisi Valle ; https://orcid.org/0000-0002-8069-6372; http://lattes.cnpq.br/9337327437538048; França, Hildegardo Seibert ; https://orcid.org/0000-0001-6129-8793; http://lattes.cnpq.br/1284874997224988; Costa, Helcio ; https://orcid.org/; http://lattes.cnpq.br/2086091514383462; Pereira, Fausto Edmundo Lima ; http://lattes.cnpq.br/4065537941002091
Pineapple is a fruit of great economic importance in fruit farming in Brazil and of high consumption worldwide, used in human and animal food, among other uses. Among the phytosanitary factors limiting the crop, fusariosis stands out, a disease caused by the fungus Fusarium guttiforme, which compromises the quality of the fruits for the market and produces an emerging mycotoxin called beauvericin (BEA), potentially harmful to the health of consumers. The importance of accurate and validated techniques for detecting BEA and establishing maximum allowable limits is urgent, as there is still no regulation in legislation in any country. Analytical techniques for the detection of mycotoxins have proven to be effective, with few studies for the detection of BEA in pineapple. Reliability in detection allows careful selection of evaluated products, ensuring food safety and economic responsibility. High Performance Liquid Chromatography (HPLC), Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS) combined with the matrix-assisted laser desorption/ ionization (MALDI) source and UHPLC-MS (Ultra Efficiency Liquid Chromatograph coupled to a mass spectrometer) were used to detect BEA in this work. The rice substrate was tested for the development of F. guttiforme and production of BEA, as it is a favorable environment for the development of the fungus. The HPLC-DAD results, in the first analyses, made it possible to detect BEA in the tissues of samples visibly infected with F. guttiforme (E-514), however in samples of apparently healthy tissue the mycotoxin was not detected. With MALDI-FT-ICR MS, BEA was detected in both types of samples, showing that, in addition to the greater sensitivity of the method, there is the possibility that the mycotoxin may diffuse into the apparently healthy tissues of fruits inoculated with F. guttiforme, with these tissues also being contaminated, but at limits not detectable by HPLC-DAD. The rice substrate proved to be efficient for the detection of beauvericin produced by isolates of F. guttiforme cultivated in vitro, which allows its use to produce the mycotoxin on a larger scale for subsequent studies. After these first results, UHPLC-MS was used for further analyses, however the answers were not satisfactory. In studies for the detection of BEA in pineapples previously inoculated with F. guttiforme (E-514), the analytical method that proved to be most accurate, until regulatory limits were met, was MALDI-FT-ICR MS, showing extremely high sensitivity of the method for mycotoxin detection and the ability to detect very small quantities not detected in HPLC-DAD
Determinação do perfil molecular dos extratos de folhas de Carica papaya (L.) em busca de biomarcadores estágio-dependentes relacionados à doença da meleira do mamoeiro
(Universidade Federal do Espírito Santo, 2024-07-12) Britto, Isabella Oliveira; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/; https://orcid.org/; http://lattes.cnpq.br/; Leite, João Paulo Viana; https://orcid.org/; http://lattes.cnpq.br/; Silva, Diolina Moura ; https://orcid.org/; http://lattes.cnpq.br/; Gonçalves, Juliana Barbosa Coitinho; https://orcid.org/; http://lattes.cnpq.br/; Lima, Graziela Domingues de Almeida; http://lattes.cnpq.br/
Papaya Sticky Disease is caused by PMeV viral complex and poses a significant threat to papaya production worldwide. Infected plants remain asymptomatic until flowering and fructification, acting as silent reservoirs of the virus in the field. Secondary metabolites could act as potential biomarkers of disease progression and can be detected using chromatographic techniques for early disease diagnosis. In this study, molecular profiles of ethanolic extracts from C. papaya leaves in pre- and postflowering (2.5, 5.0, and 7.5 mg.mL-1) and fractions were evaluated by UV-Vis spectroscopy, reversed-phase chromatography, ion exchange chromatography, and mass spectrometry. The 7.5 mg.mL-1 concentration was selected as the experimental concentration for crude extracts. Chromatograms showed symmetrical peaks, eluted within similar retention time ranges across the three plant stages, with narrow base widths, similar peak shapes, and no tailing, suggesting the presence of a limited number of potentially isolable compounds. Analytical parameters indicated quantitatively larger chromatographic peaks in pre-flowering leaf extracts compared to post-flowering ones, reflecting a possible defense response against viral infection. Significant differences in chromatographic profiles between the different plant stages were observed, indicating the potential of these groups as biomarkers for Papaya Sticky Disease. This study developed a sensitive and reproducible chromatographic method to distinguish plant stages, identifying flavonoids and alkaloids as components of strategic groups related to the defense system against viral infections.

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