GENOTOXICITY OF SHALLOW WATERS NEAR THE BRAZILIAN ANTARCTIC STATION "COMANDANTE FERRAZ" (EACF), ADMIRALTY BAY, KING GEORGE ISLAND, ANTARCTICA

Series of biomonitoring surveys were undertaken weekly in February 2012 to investigate the genotoxicity of the shallow waters around the Brazilian Antarctic Station "Comandante Ferraz" (EACF). The comet assay was applied to assess the damage to the DNA of hemocytes of the crustacean amphipods Gondogeneia antarctica collected from shallow waters near the Fuel Tanks (FT) and Sewage Treatment Outflow (STO) of the research station, and compare it to the DNA damage of animals from Punta Plaza (PPL) and Yellow Point (YP), natural sites far from the EACF defined as experimental controls. The damage to the DNA of hemocytes of G. antarctica was not significantly different between sites in the biomonitoring surveys I and II. In survey III, the damage to the DNA of animals captured in shallow waters near the Fuel Tanks (FT) and Sewage Treatment Outflow (STO) was significantly higher than that of the control site of Punta Plaza (PPL). In biomonitoring survey IV, a significant difference was detected only between the FT and PPL sites. Results demonstrated that the shallow waters in front of the station may be genotoxic and that the comet assay and hemocytes of G. antarctica are useful tools for assessing genotoxicity in biomonitoring studies of Antarctic marine coastal habitats.Séries de biomonitoramentos foram executadas semanalmente, durante o mês de fevereiro de 2012, para se investigar a genotoxicidade de ambientes costeiros rasos no entorno da Estação Antártica "Comandante Ferraz (EACF). O ensaio cometa foi aplicado para se avaliar os danos ao DNA hemocitário de crustáceos anfípodes Gondogeneia antarctica coletados em áreas rasas próximas aos tanques de armazenamento de combustível (FT) e saída de efluentes da estação de tratamento de esgoto (STO), em comparação aos danos ao DNA de animais provenientes de Punta Plaza (PPL) e Yellow Point (YP), locais naturais distantes da EACF, definidos como controles experimentais. O dano ao DNA hemocitário de G. antarctica não foi significativamente diferente entre locais, nos biomonitoramentos I e II. No biomonitoramento III, o dano ao DNA de animais coletados em águas rasas próximas aos tanques de combustível e saída de efluentes de esgoto foi significativamente maior do que aquele do controle de Punta Plaza. No biomonitoramento IV, a diferença foi significativa somente entre os locais dos tanques de combustível e de Punta Plaza. Estes resultados demonstram que a contaminação das águas em frente à EACF pode ser genotóxica e que, tanto o ensaio cometa quanto os hemócitos de G. antarctica são ferramentas úteis na avaliação da genotoxicidade em estudos de biomonitoramento de habitats marinhos costeiros da Antártica

The Cell Wall Integrity Pathway Contributes to the Early Stages of Aspergillus Fumigatus Asexual Development

Aspergillus fumigatus is a major cause of human disease. The survival of this fungus is dependent on the cell wall organization and function of its components. The cell wall integrity pathway (CWIP) is the primary signaling cascade that controls de novo synthesis of the cell wall in fungi. Abundant conidiation is a hallmark in A. fumigatus, and uptake of conidia by a susceptible host is usually the initial event in infection. The formation of conidia is mediated by the development of fungus-specific specialized structures, conidiophores, which are accompanied by cell wall remodeling. The molecular regulation of these changes in cell wall composition required for the rise of conidiophore from the solid surface and to disperse the conidia into the air is currently unknown. Here, we investigated the role of CWIP in conidiation. We show that CWIP pkcAG579R, ΔmpkA, and ΔrlmA mutants displayed reduced conidiation during synchronized asexual differentiation. The transcription factor RlmA directly regulated the expression of regulators of conidiation, including flbB, flbC, brlA, abaA, and rasB, as well as genes involved in cell wall synthesis and remodeling, and this affected the chitin content in aerial hyphae. Phosphorylation of RlmA and MpkA was increased during asexual differentiation. We also observed that MpkA physically associated with the proteins FlbB, FlbC, BrlA, and RasB during this process, suggesting another level of cross talk between the CWIP and asexual development pathways. In summary, our results support the conclusion that one function of the CWIP is the regulation of asexual development in filamentous fungi. IMPORTANCE A remarkable feature of the human pathogen Aspergillus fumigatus is its ability to produce impressive amounts of infectious propagules known as conidia. These particles reach immunocompromised patients and may initiate a life-threatening mycosis. The conidiation process in Aspergillus is governed by a sequence of proteins that coordinate the development of conidiophores. This process requires the remodeling of the cell wall so that the conidiophores can rise and withstand the chains of conidia. The events regulating cell wall remodeling during conidiation are currently unknown. Here, we show that the cell wall integrity pathway (CWIP) components RlmA and MpkA directly contribute to the activation of the conidiation cascade by enabling transcription or phosphorylation of critical proteins involved in asexual development. This study points to an essential role for the CWIP during conidiation and provides further insights into the complex regulation of asexual development in filamentous fungi.Fundação de Amparo à Pesquisa do Estado de São Paulo 2015/17541-0, 2016/07870-9, 2017/19694-3Conselho Nacional de Desenvolvimento Científico e Tecnológico 462383/2014-

Citogenotoxic response of juvenile cobia Rachycentron canadum (Linnaeus, 1766) reared in two different systems

A produção da aquicultura vem crescendo em todo o mundo e o cultivo de peixes marinhos no Brasil é ainda muito recente. Condições intensivas de aquicultura podem causar estresse fisiológico ao organismo do cultivo, o que pode ser avaliado por biomarcadores citogenotóxicos. O objetivo desse estudo foi avaliar o efeito genotóxico das condições de cultivo nos eritrócitos de juvenis de beijupirás Rachycentron canadum usando o ensaio cometa e o ensaio de micronúcleo e outras anormalidades nucleares. Juvenis de beijupirá foram cultivados por 13 semanas em um tanque indoor com circulação aberta de água e em um tanque-rede. O ensaio comera e o ensaio de anormalidades nucleares detectaram um maior dano ao DNA e uma maior frequência de anormalidades nucleares em eritrócitos de peixes cultivados no tanque indoor. Results showed that two methods are complementary. Além disso, beijupirás foram injetados com β-naftoflavona (BNF) nas concentrações de 2mgkg-1 e 10mgkg-1 em condições controladas de laboratório e mantidos por 7 dias em tanques separados para um melhor entendimento dos mecanismos de resposta dessa espécie a uma substância tóxica. O ensaio cometa não detectou nenhuma diferença significativa entre os peixes injetados com BNF e os da condição controle, enquanto que o ensaio das anormalidades nucleares apresentaram diferenças significativas entre peixes injetados com BNF e os do controle. Os danos identificados pelo ensaio cometa são quebras reparáveis na fita do DNA, enquanto que as anormalidades nucleares são permanentes. Possivelmente, o período de de manutenção dos peixes nos tanques após a injeção foi suficiente para limpar a BNF dos organismos e reparar as quebras na fita do DNA. Como os beijupirás parecem responder muito bem a compostos genotóxicos, o ensaio cometa e o ensaio de anormalidades nucleares podem ser ferramentas úteis para monitorar as condições de cultivo.Aquaculture production is continuously growing worldwide, and marine fish farming in Brazil is still in its infancy. Intensive farming conditions may cause physiological stress to the cultured organism, which can be evaluated by citogenotoxic biomarkers. The aim of this study was to assess the genotoxic effect of the rearing conditions in red blood cells of juvenile cobia Rachycentron canadum by using comet assay and micronucleus and other nuclear abnormalities assay. Juvenile cobia were reared for 13 weeks in indoor tank with open water circulation and in near shore cage. The comet assay and the nuclear abnormalities assay detected higher DNA damage and higher nuclear abnormalities frequency in erythrocytes of fish reared in the indoor tank. Results showed that two methods are complementary. Additionally, cobia were injected with ß-naphthoflavone (BNF) at concentrations of 2mgkg-1 and 10mgkg-1 in laboratory controlled conditions, and maintained for 7 days in separate tanks to better understand the response mechanisms of this species to a toxic substance. The comet assay did not detect any significant differences between BNF injected and control fish, whereas nuclear abnormalities assay showed significant differences between BNF injected and the control groups. The damages identified by the comet assay are repairable breaks in the DNA strands, whereas nuclear abnormalities may be permanent. Possibly the period of maintenance after injection was enough to clean BNF from the organisms and to repair the breaks in the DNA strands. As cobia seems to respond very well to genotoxic elements, comet assay and nuclear abnormalities assay would be useful tools to monitor farming conditions

The Aspergillus nidulans pyruvate dehydrogenase kinases are essential to integrate carbon source metabolism

The pyruvate dehydrogenase complex (PDH), that converts pyruvate to acetyl-coA, is regulated by pyruvate dehydrogenase kinases (PDHK) and phosphatases (PDHP) that have been shown to be important for morphology, pathogenicity and carbon source utilization in different fungal species. The aim of this study was to investigate the role played by the three PDHKs PkpA, PkpB and PkpC in carbon source utilization in the reference filamentous fungus Aspergillus nidulans, in order to unravel regulatory mechanisms which could prove useful for fungal biotechnological and biomedical applications. PkpA and PkpB were shown to be mitochondrial whereas PkpC localized to the mitochondria in a carbon source-dependent manner. Only PkpA was shown to regulate PDH activity. In the presence of glucose, deletion of pkpA and pkpC resulted in reduced glucose utilization, which affected carbon catabolite repression (CCR) and hydrolytic enzyme secretion, due to de-regulated glycolysis and TCA cycle enzyme activities. Furthermore, PkpC was shown to be required for the correct metabolic utilization of cellulose and acetate. PkpC negatively regulated the activity of the glyoxylate cycle enzyme isocitrate lyase (ICL), required for acetate metabolism. In summary, this study identified PDHKs important for the regulation of central carbon metabolism in the presence of different carbon sources, with effects on the secretion of biotechnologically important enzymes and carbon source-related growth. This work demonstrates how central carbon metabolism can affect a variety of fungal traits and lays a basis for further investigation into these characteristics with potential interest for different applications.We would like to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant numbers 2016/07870-9 for GHG and 2014/00789-6 for LJA), the Science Foundation Ireland (SCI, grant number 13/CDA/2142 for OB) for providing financial support.info:eu-repo/semantics/publishedVersio

The Brazilian Ceremony in Honor of Body Donors: An opportunity to express gratitude and reflect on medical education

Introduction: Memorial ceremonies are a way for institutions to show their gratitude for the gesture made by body donors and are an opportunity for students to reflect on the ethical paradigm shift. Therefore, this study aims to describe the memorial ceremony at the Federal University of Health Sciences of Porto Alegre and assess the perception of both students and the donors’ relatives regarding the event. Material and methods: In 2016, a questionnaire was applied to donors’ relatives with questions regarding structure of the ceremony, ethical training of the students and importance of body donation. Another questionnaire was applied to the first-year students of the medical school, with questions about understanding the use of bodies, and their thoughts on death and relevant ethical issues. Results: Fifty questionnaires were answered by family members and 98% agreed that the ceremony was an opportunity to feel closer to the donors and 100% said that the ceremony is important in the ethical training of students. Among students, 68 questionnaires were answered, and all affirmed that the event had a positive impact on the perception of physician-patient relationship and personal growth, and 86.7% reported enhanced empathy. Conclusion: The memorial ceremony seems to encourage students to reflect on ethical issues, because at the same time that the donors’ relatives perceive an attitude of care and respect on the part of the students, the students state a heightened sense of commitment and responsibility regarding their learning and professional formation, which depends on the gesture made by the donors

Prevalence of constipation in Down syndrome and association with behavioral problems

Objetivo: Investigar hábitos intestinais de crianças, jovens e adultos com Síndrome de Down (SD) para identificação da prevalência de constipação intestinal e analisar possíveis relações entre constipação e problemas de comportamento. Métodos: Estudo descritivo, exploratório e comparativo. Participaram 130 pessoas com diagnóstico médico de SD, idades entre sete meses e 50 anos, sem cardiopatias não corrigidas ou doenças intestinais. A avaliação foi realizada com um questionário sobre hábitos intestinais, criado a partir dos Critérios de Roma IV e das recomendações fornecidas pela Organização Mundial de Gastroenterologia e outras Associações Médicas Norte-americanas e Europeias, Escala de Bristol e Questionário de Capacidades e Dificuldades (SDQ). Resultados: Verificou-se 43 participantes constipados (33,1%). A prevalência por faixa etária foi 47% (0-35 meses), 40% (3-5 anos), 22% (6-11 anos), 28% (12-19 anos) e 17% (> 20 anos), sem diferenças em função do sexo. Os participantes constipados obtiveram escores mais elevados para Hiperatividade, bem como maior frequência de escores desviantes para Problemas de Conduta e Problemas Totais. Conclusões: A constipação intestinal dentre as pessoas com SD apresentou maior prevalência nos participantes mais novos. A maior presença de problemas de comportamento dentre os constipados sugere que o funcionamento intestinal está relacionado com as dificuldades comportamentais.Objective: To investigate intestinal habits of children, adolescents and adults with Down Syndrome (DS) in order to identify the prevalence of intestinal constipation and to analyze possible relationships between constipation and behavior problems. Methods: Descriptive, exploratory and comparative study. Participated 130 people with medical diagnosis of DS, aged between seven months to fifty years old, without uncorrected heart diseases or diagnosed intestinal diseases. The assessment was conducted using a questionnaire on intestinal habits, based on the Rome IV criteria and on the recommendations provided by the World Gastroenterology Organization, and other North American and European Associations; Bristol Scale and Strengths and Difficulties Questionnaire (SDQ). Results: 43 constipated participants (33.1%). The prevalence rate by age group was 47% (0-35 months), 40% (3-5 years), 22% (6-11 years), 28% (12-19 years) e 17% (> 20 years), without any sex difference. Constipated participants obtained significantly higher scores on the SDQ Hyperactivity scale, as well as more deviant scores on the Conduct Problems and Total Problems scales. Conclusions: Constipation among people with DS was more prevalent in younger participants. The greater presence of behavior problems among constipated individuals suggests that intestinal functioning is related to behavioral difficulties

The AGC kinase YpkA regulates sphingolipids biosynthesis and physically interacts with SakA MAP kinase in Aspergillus fumigatus

Sphingolipids (SL) are complex lipids and components of the plasma membrane which are involved in numerous cellular processes, as well as important for virulence of different fungal pathogens. In yeast, SL biosynthesis is regulated by the "AGC kinases" Ypk1 and Ypk2, which also seem to connect the SL biosynthesis with the cell wall integrity (CWI) and the High Osmolarity Glycerol (HOG) pathways. Here, we investigate the role of ypkA(YPK1) in SL biosynthesis and its relationship with the CWI and the HOG pathways in the opportunistic human pathogen Aspergillus fumigatus. We found that ypkA is important for fungal viability, since the 1 ypkA strain presented a drastically sick phenotype and complete absence of conidiation. We observed that under repressive condition, the conditional mutant niiA::ypkA exhibited vegetative growth defects, impaired germination and thermosensitivity. In addition, the ypkA loss of function caused a decrease in glycosphingolipid (GSL) levels, especially the metabolic intermediates belonging to the neutral GSL branch including dihydroceramide (DHC), ceramide (Cer), and glucosylceramide (GlcCer), but interestingly a small increase in ergosterol content. Genetic analyzes showed that ypkA genetically interacts with the MAP kinases of CWI and HOG pathways, mpkA and sakA, respectively, while only SakA physically interacts with YpkA. Our results suggest that YpkA is important for fungal survival through the regulation of GSL biosynthesis and cross talks with A. fumigatus MAP kinase pathways9CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP462383/2014-82009/53546-5; 2015/17541-0; 2017/19694-3This study was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP Grant Numbers: 2009/53546-5, 2015/17541-0, and 2017/19694-3) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Grant Number 462383/2014-8), Brazil to IM. This work was also supported by NIH grants (AI116420 and AI125770) and by the VA Merit Award grant (I01BX002624) to MDP. The funding body had no role in designing the study or in collecting, analyzing and interpreting the data, or in writing the manuscrip

Conservation of wild mushrooms through electron beam irradiation

The high perishability is a characteristic of the mushrooms consumed in fresh. Therefore, it is mandatory the application of effective conservation technologies to preserve and protect their chemical composition and nutritional value. Drying processes are widely used, but do not avoid the development of bacteria and fungi which have the ability to survive for long periods of time in dry foods, causing the loss of some nutrients and leading to food browning and oxidation of lipids and vitamins [1]. Irradiation appears as an alternative to food preservation assuring and maintaining its quality [2]. In this work, the effects of electron beam irradiation and storage time on nutritional and chemical parameters of wild samples of Macrolepiota procera (Scop.) Singer, previously submitted to a drying process (oven at 30 ºC), were assessed. The wild mushroom samples were collected in Trás-os-Montes; electron beam irradiation (doses 0.5, 1 and 6 kGy) was carried out in the INCT- Institute of Nuclear Chemistry and Technology in Warsaw, Poland and the analyses were performed over the storage period (0, 6 and 12 months). The results were compared with a control (non-irradiated samples). The nutritional value was determined according to the official procedures of food analysis, while the profiles of fatty acids, tocopherols, mono and oligosaccharides were obtained by chromatographic techniques [1]. The irradiation showed a better capacity to maintain the nutritional and chemical profile, in comparison with the storage time. Effectively, the storage time had a significant effect in all parameters, but fatty acids undergone significant changes both with irradiation doses and storage time. Electron beam irradiation can be considered a suitable technique for conservation of mushrooms for long periods of time, attenuating the changes caused by the drying treatment.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), A. Fernandes (SFRH/BPD/114753/2016) and J.C.M. Barreira contract.info:eu-repo/semantics/publishedVersio

Knowledge and interest in medical ethics and bioethics in medical graduation

OBJECTIVES: To evaluate the interest and knowledge about Medical Ethics and Bioethics in medical graduation. METHODS: Transversal and descriptive study. Two different questionnaires were applied with questions about the interest in Medical Ethics and Bioethics and the knowledge about the Brazilian Code of Medical Ethics (CME), one to professors and the other to students. RESULTS: One hundred and one professors and 331 students answered the questionnaires. The Brazilian CME had been read by 86.2% of the teachers and by 100% of the students. The importance given to the discipline Medical Ethics, on a scale from 1 to 5, was similar among teachers and students (4.7 ± 0.7 vs. 4.5 ± 0.8; p = 0.086); however the self-evaluation on knowledge about this subject was higher in the first group (3.4 ± 0.9 vs. 3.2 ± 0.7; p = 0.017). In a block with 9 questions, the right answer was given by 5,0 ± 1.9 of teachers and 5.9 ± 1.5 of students (p < 0,001); the mean of correct answers were related to the reading of the CME. CONCLUSIONS: The present study presents unpublished data about the perception of medical teachers and students about medical ethics and bioethics and can be useful for improving the teaching of these disciplines in our medical schools.OBJETIVOS: Avaliar o interesse e o conhecimento sobre ética médica e bioética na graduação médica. MÉTODOS: Estudo transversal e descritivo. Foram utilizados dois questionários auto-aplicáveis, um para docentes e outro para discentes, com questões sobre o interesse em ética médica e bioética e conhecimento sobre o Código de Ética Médica (CEM). RESULTADOS: Foram avaliados 101 professores e 331 estudantes. O CEM brasileiro foi lido por 86,2% dos professores e 100% dos alunos. A importância dada à disciplina de ética médica, numa escala de 1 a 5, foi semelhante entre professores e estudantes (4,7 ± 0,7 vs. 4,5 ± 0,8; p = 0,086), enquanto o autoconhecimento sobre o tema foi maior no primeiro grupo (3,4 ± 0,9 vs. 3,2 ± 0,7; p = 0,017). De um total de nove questões avaliadas sobre o conhecimento do CEM, a média de acertos foi de 5,0 ± 1,9 questões para os professores e de 5,9 ± 1,5 para os acadêmicos (p < 0,001), sendo os acertos correlacionados positivamente com a leitura do código. CONCLUSÕES: Este estudo fornece um panorama inédito sobre a percepção de professores e estudantes de Medicina sobre a ética médica e bioética, podendo servir para fundamentar a melhora do ensino dessa disciplina em nossas faculdades.Universidade Federal da BahiaEscola Bahiana de Medicina e Saúde PúblicaUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Hospital São PauloInstituto do coração da Bahia Hospital Ana NeryUniversidade Federal da Bahia Hospital Universitário Professor Edgar SantosUNIFESP, EPM, Hospital São PauloSciEL

Dysbiotic oral microbiota contributes to alveolar bone loss associated with obesity in mice

Periodontal diseases (PD) are inflammatory conditions that affect the teeth supporting tissues. Increased body fat tissues may contribute to activation of the systemic inflammatory response, leading to comorbidities. Some studies have shown that individuals with obesity present higher incidence of PD than eutrophics. Objective: To investigate the impact of obesity on periodontal tissues and oral microbiota in mice. Methodology: Two obesity mice models were performed, one using 12 weeks of the dietary protocol with a high-fat (HF) diet in C57BL/6 mice and the other using leptin receptor-deficient mice (db/db-/-), which became spontaneously obese. After euthanasia, a DNA-DNA hybridization technique was employed to evaluate the microbiota composition and topical application of chlorhexidine (CHX), an antiseptic, was used to investigate the impact of the oral microbiota on the alveolar bone regarding obesity. Results: Increased adipose tissue may induce alveolar bone loss, neutrophil recruitment, and changes in the oral biofilm, similar to that observed in an experimental model of PD. Topical application of CHX impaired bone changes. Conclusion: Obesity may induce changes in the oral microbiota and neutrophil recruitment, which are associated with alveolar bone loss