Modelling the interaction of steroid receptors with endocrine disrupting chemicals

BACKGROUND: The organic polychlorinated compounds like dichlorodiphenyltrichloroethane with its metabolites and polychlorinated biphenyls are a class of highly persistent environmental contaminants. They have been recognized to have detrimental health effects both on wildlife and humans acting as endocrine disrupters due to their ability of mimicking the action of the steroid hormones, and thus interfering with hormone response. There are several experimental evidences that they bind and activate human steroid receptors. However, despite the growing concern about the toxicological activity of endocrine disrupters, molecular data of the interaction of these compounds with biological targets are still lacking. RESULTS: We have used a flexible docking approach to characterize the molecular interaction of seven endocrine disrupting chemicals with estrogen, progesterone and androgen receptors in the ligand-binding domain. All ligands docked in the buried hydrophobic cavity corresponding to the hormone steroid pocket. The interaction was characterized by multiple hydrophobic contacts involving a different number of residues facing the binding pocket, depending on ligands orientation. The EDC ligands did not display a unique binding mode, probably due to their lipophilicity and flexibility, which conferred them a great adaptability into the hydrophobic and large binding pocket of steroid receptors. CONCLUSION: Our results are in agreement with toxicological data on binding and allow to describe a pattern of interactions for a group of ECD to steroid receptors suggesting the requirement of a hydrophobic cavity to accommodate these chlorine carrying compounds. Although the affinity is lower than for hormones, their action can be brought about by a possible synergistic effect

Solving biclustering with a GRASP-like metaheuristic: two case-study on gene expression analysis

The explosion of "omics" data over the past few decades has generated an increasing need of efficiently analyzing high-dimensional gene expression data in several different and heterogenous contexts, such as for example in information retrieval, knowledge discovery, and data mining. For this reason, biclustering, or simultaneous clustering of both genes and conditions has generated considerable interest over the past few decades. Unfortunately, the problem of locating the most significant bicluster has been shown to be NP-complete. We have designed and implemented a GRASP-like heuristic algorithm to efficiently find good solutions in reasonable running times, and to overcome the inner intractability of the problem from a computational point of view. Experimental results on two datasets of expression data are promising indicating that this algorithm is able to find significant biclusters, especially from a biological point of view

Profiling the Course of Resolving vs. Persistent Inflammation in Human Monocytes: The Role of IL-1 Family Molecules

Monocytes and macrophages have a central role in all phases of an inflammatory reaction. To understanding the regulation of monocyte activation during a physiological or pathological inflammation, we propose two in vitro models that recapitulate the different phases of the reaction (recruitment, initiation, development, and resolution vs. persistence of inflammation), based on human primary blood monocytes exposed to sequential modifications of microenvironmental conditions. These models exclusively describe the functional development of blood-derived monocytes that first enter an inflammatory site. All reaction phases were profiled by RNA-Seq, and the two models were validated by studying the modulation of IL-1 family members. Genes were differentially modulated, and distinct clusters were identified during the various phases of inflammation. Pathway analysis revealed that both models were enriched in pathways involved in innate immune activation. We observe that monocytes acquire an M1-like profile during early inflammation, and switch to a deactivated M2-like profile during both the resolving and persistent phases. However, during persistent inflammation they partially maintain an M1 profile, although they lose the ability to produce inflammatory cytokines compared to M1 cells. The production of IL-1 family molecules by ELISA reflected the transcriptomic profiles in the distinct phases of the two inflammatory reactions. Based on the results, we hypothesize that persistence of inflammatory stimuli cannot maintain the M1 activated phenotype of incoming monocytes for long, suggesting that the persistent presence of M1 cells and effects in a chronically inflamed tissue is mainly due to activation of newly incoming cells. Moreover, being IL-1 family molecules mainly expressed and secreted by monocytes during the early stages of the inflammatory response (within 4-14 h), and the rate of their production decreasing during the late phase of both resolving and persistent inflammation, we suppose that IL-1 factors are key regulators of the acute defensive innate inflammatory reaction that precedes establishment of longer-term adaptive immunity, and are mainly related to the presence of recently recruited blood monocytes. The well-described role of IL-1 family cytokines and receptors in chronic inflammation is therefore most likely dependent on the continuous influx of blood monocytes into a chronically inflamed site

USE OF SYSTEMIC FUNGICIDES COMBINED WITH MULTISITE TO CONTROL OF ASIAN RUST AND SOYBEAN YIELD

Asian soybean rust is an aggressive disease and chemical control must be handled assertively to minimize productivity damage. The objective was to evaluate systemic fungicides and multisite mancozeb, combined or not, and the effects on disease control and soybean yield, cv. 'BMX Lança'. The work was conducted in the field in the 2016/17 and 2017/18 crops, in a randomized complete block design, with four replications. The following treatments were evaluated: T+P) trifloxystrobin + prothioconazole; A+B) azoxystrobin + benzovindiflupyr; T+P+MB) trifloxystrobin + prothioconazole + mancozeb; A+B+MB) azoxystrobin + benzovindiflupyr + mancozeb; MB) mancozeb; and Control (without fungicide application). After the occurrence of the first symptoms of the disease, the severity (%) was evaluated in five trifolia per plot, every seven days, making it possible to calculate the area under the disease progress curve (AUDPC). The yield components and productivity (kg ha-1) were also evaluated. In both crops, the combination of systemic fungicides and mancozeb reduced the severity (%) and progress of Asian rust, reaching a control above 81%. Treatment with trifloxystrobin + prothioconazole + mancozeb (T+P+MB) obtained the highest percentages of control (94.5% - 2016/17, and 93.7% - 2017/18) of Asian rust, and provided, in the 2016/17 crop, productivity at 15 sc ha-1, in relation to the fungicide applied alone (T+P). The combination of systemic fungicides and mancozeb, as proposed in this work, reduces the severity and progress of Asian rust and ensures the maintenance of the productive potential of the soybean.A ferrugem asiática da soja é uma doença agressiva e o controle químico deve ser manejado assertivamente, para minimizar danos sobre a produtividade. Objetivou-se avaliar fungicidas sistêmicos e o multissítio mancozeb, combinados ou não, e os efeitos sobre o controle da doença e a produtividade da soja, cv. ‘BMX Lança’. O trabalho foi conduzido à campo nas safras 2016/17 e 2017/18, em delineamento de blocos ao acaso, com quatro repetições. Foram avaliados os tratamentos: T+P) trifloxistrobina + protioconazol; A+B) azoxistrobina + benzovindiflupyr; T+P+MB) trifloxistrobina + protioconazol + mancozeb; A+B+MB) azoxistrobina + benzovindiflupyr + mancozebe; MB) mancozeb; e Testemunha (sem aplicação de fungicidas). Após o surgimento dos primeiros sintomas da doença, a severidade (%) foi avaliada em cinco trifólios por parcela, a cada sete dias, possibilitando o cálculo da área abaixo da curva de progresso da doença (AACPD). Avaliou-se também os componentes de rendimento e a produtividade (kg ha-1). Em ambas as safras, a combinação entre fungicidas sistêmicos e o mancozeb reduziu a severidade (%) e o progresso de ferrugem asiática, atingindo um controle acima de 81%. O tratamento com trifloxistrobina + protioconazol + mancozeb (T+P+MB) obteve os maiores percentuais de controle (94,5% - 2016/17; e 93,7% - 2017/18) da ferrugem asiática, e proporcionou, na safra 2016/17, produtividade superior em 15 sc ha-1, em relação ao fungicida aplicado isoladamente (T+P). A combinação entre fungicidas sistêmicos e mancozeb, conforme proposto nesse trabalho, reduz a severidade e o progresso da ferrugem asiática e assegura a manutenção do potencial produtivo da soja

Human mitochondrial 5'-deoxyribonucleotidase. Overproduction in cultured cells and functional aspects.

Deoxynucleoside triphosphates (dNTPs) used for mitochondrial DNA replication are mainly formed by phosphorylation of deoxynucleosides imported into mitochondria from the cytosol. We earlier obtained evidence for a mitochondrial 5′-nucleotidase (dNT2) with a pronounced specificity for dUMP and dTMP and suggested that the enzyme protects mitochondrial DNA replication from excess dTTP. In humans, accumulation of dTTP causes a mitochondrial genetic disease. We now establish that dNT2 in vivo indeed is located in mitochondria. The native enzyme shows the same substrate specificity and affinity for inhibitors as the recombinant dNT2. We constructed ponasterone-inducible cell lines overproducing dNT2 with and without the green fluorescent protein (GFP) linked to its C terminus. The fusion protein occurred in mitochondria mostly in an inactive truncated form, with only a short C-terminal fragment of dNT2 linked to GFP. No truncation occurred when dNT2 and GFP were not linked. The cell mitochondria then contained a large excess of active dNT2 with or without the mitochondrial presequence. After removal of ponasterone overproduced dNT2 disappeared only slowly from the cells, whereas dNT2-mRNA was lost rapidly. Overproduction of dNT2 did not lead to an increased excretion of pyrimidine deoxyribonucleosides, in contrast to overproduction of the corresponding cytosolic deoxynucleotidase, suggesting that the mitochondrial enzyme does not affect overall cellular deoxynucleotide turnover

Caracterização molecular e grupos de compatibilidade vegetativa de Fusarium spp. patogênicos à Ilex paraguariensis A. St.-Hil.

In the central-western and southern regions of Brazil, the cultivation of yerba mate (Ilex paraguariensis) is considered an important source of forest extraction, besides providing an alternative income, especially for families. The yerba mate, like other forest species, is infected by several pathogens, such as fungi of the genus Fusarium, causing root rot. Pathogens have wide genetic variability, even within a single genus and species, resulting from adaptations to their environments. This study aimed at characterizing molecularly isolated Fusarium spp. through the sequencing of the ITS and β-tubulin genome regions, identifying the species level as well as the formation of vegetative compatibility groups (VCG) among these isolates of the root rot pathogen in yerba mate.  The sequencing of the ITS and β-tubulin regions, defined the separation between the seven pathogenic isolates of F. solani and F. oxysporum. For the same isolates, the technique to obtain nit mutants was suitable for the phenotypic classification, obtaining five VCGs for species of Fusarium, root rot in yerba mate.Nas regiões Centro-Oeste e Sul do Brasil, a cultura da erva-mate (Ilex paraguariensis) constitui-se uma importante fonte de extrativismo florestal, além de diversificar a renda, principalmente na agricultura familiar. A erva-mate, como outras espécies florestais, é infectada por diversos patógenos, como o gênero Fusarium, causador da podridão-de-raízes. Os patógenos possuem ampla variabilidade genética, mesmo dentro de um único gênero e dentro de uma única espécie, resultado de adaptações ao meio em que vivem. Este estudo teve como objetivo avaliar o sequenciamento das regiões do genoma ITS e β-tubulina na identificação em nível de espécie dos isolados de Fusarium spp. patogênicos à erva-mate, e identificar a formação de grupos de compatibilidade vegetativa (VCG) entre os isolados de Fusarium spp. patogênicos à erva-mate. O sequenciamento das regiões ITS e β-tubulina consolidou a separação entre os sete isolados patogênicos em F. solani e F. oxysporum. Para esses isolados, a técnica para obtenção de mutantes nit foi adequada para a classificação fenotípica, fundamentais nos testes de VCG para espécies de Fusarium, sendo obtidos cinco VCG

Anti-aging and neuroprotective properties of Grifola frondosa and Hericium erinaceus extracts

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/nu14204368/s1. Table S1. Analytes identified by GC/MS on the basis of match with NIST2014 library and the corresponding target ions used to quantify them. Table S2. Yeast strains used in this study. Figure S1 Calibration curve and linear regression curve for ET. Figure S2 Evaluation of interference of fungal extracts on ThT assay. Figure S3. High fungal extract concentrations are toxic for adult flies.Nutrition has relevant consequences for human health and increasing pieces of evidence indicate that medicinal mushrooms have several beneficial effects. One of the main issues in Western countries is represented by the challenges of aging and age-related diseases, such as neurodegenerative disorders. Among these, Parkinson’s disease (PD) affects 10 million people worldwide and is associated with α-synuclein misfolding, also found in other pathologies collectively called synucleinopathies. Here, we show that aqueous extracts of two edible mushrooms, Grifola frondosa and Hericium erinaceus, represent a valuable source of β-glucans and exert anti-aging effects in yeast. Their beneficial effects are mediated through the inhibition of the Ras/PKA pathway, with increased expression of heat shock proteins, along with a consistent increase of both mean and maximal lifespans. These fungal extracts also reduce the toxicity of α-synuclein heterologously expressed in yeast cells, resulting in reduced ROS levels, lower α-synuclein membrane localization, and protein aggregation. The neuroprotective activity of G. frondosa extract was also confirmed in a PD model of Drosophila melanogaster. Taken together, our data suggest the use of G. frondosa and H. erinaceus as functional food to prevent aging and age-related disorders, further supporting the neuro-healthy properties of these medicinal mushroom extracts.We acknowledge financial support from the Italian Ministry of University and Research (MUR) through grant “Dipartimenti di Eccellenza 2017” to University of Milano-Bicocca, Department of Biotechnology and Biosciences. This research was also supported by the Italian Ministry of University and Research (MUR): Dipartimenti di Eccellenza Program (2018–2022) Dept. of Biology and Biotechnology “L. Spallanzani”, and Fondo di Ricerca e Giovani (FRG, University of Pavia). M.L. was supported by a fellowship from Fondazione Umberto Veronesi. R.M. was supported by a fellowship from the Italian Ministry of University and Research (MUR)

Value of staging squamous cell carcinoma of the anal margin and canal using the sentinel lymph node procedure: an update of the series and a review of the literature.

BACKGROUND: Inguinal metastases in patients affected by anal cancer are an independent prognostic factor for local failure and overall mortality. Since 2001, sentinel lymph node biopsy was applied in these patients. This original study reports an update of personal and previous published series, which were compared with Literature to value the incidence of inguinal metastases T-stage related and the overall incidence of false negative inguinal metastases at sentinel node. METHODS: In all, 63 patients diagnosed with anal cancer submitted to inguinal sentinel node. Furthermore a research in the Pub Med database was performed to find papers regarding this technique. RESULTS: In our series, detection rate was 98.4%. Inguinal metastases were evidentiated in 13 patients (20.6%). Our median follow-up was 35 months. In our series, no false negative nodes were observed. CONCLUSION: Sentinel node technique in the detection of inguinal metastases in patients affected by anal cancer should be considered as a standard of care. It is indicated for all T stages in order to select patients to be submitted to inguinal radiotherapy, avoiding related morbidity in negative ones. An overall 3.7% rate of false negative must be considered acceptable

Formação continuada dos professores das escolas técnicas da Universidade Federal de Santa Maria

This article highlights the importance of the presence of teachers in continuing education courses, aiming to analyze the participation of teachers of Polytechnic and Technical Industrial - Ctism - Colleges of Federal University of Santa Maria - UFSM - in continuing education courses. For this, we conducted a quantitative survey and the data collected through a multiple choice questionnaire that was applied to teachers in these institutions. It was concluded that most teachers of Polytechnic and Ctism Colleges of UFSM participate in continuing education courses and institutions to receive this incentive. Still, teachers attend refresher courses in education, even those that are embedded in more technical areas.Key-words: professional development, technological education.Neste texto destaca-se a importância da formação continuada de professores, tendo como objetivo analisar a participação dos docentes dos Colégios Politécnico e Técnico Industrial - Ctism - da Universidade Federal de Santa Maria - UFSM - em cursos de formação continuada. Para isso, foi realizada uma pesquisa quantitativa, sendo os dados coletados por meio de um questionário de múltipla escolha que foi aplicado para os docentes das referidas instituições. Concluiu-se que a maioria dos professores dos colégios Politécnico e Ctism da UFSM participam de cursos de formação continuada e recebem incentivos das suas instituições. Ainda, os professores frequentam cursos de atualização na área da educação, mesmo aqueles que estão inseridos em áreas mais técnicas.Palavras-chave: atualização profissional, ensino tecnológico

Número de aplicações de fungicida e comparação entre cultivares com e sem tolerância no controle da ferrugem asiática da soja

A ferrugem asiática da soja (Phakopsora pachyrhizi Sydow) pode implicar em perda de toda a lavoura e, por isso, os controles genético e químico são medidas eficientes no manejo integrado dessa doença. Objetivou-se quantificar o dano e o controle da ferrugem asiática, em duas cultivares de soja, frente ao número de aplicações de fungicida (tebuconazol + picoxistrobina, 60 + 100 g i.a ha-1) nas cvs. TMG 7262 (Inox) e BMX Tornado, com e sem tolerância à doença, respectivamente. O delineamento utilizado foi de blocos casualizados com parcelas subdivididas e quatro repetições. Nas parcelas foram dispostos os tratamentos: uma (1) aplicação de fungicida (estádio R1); duas (2) aplicações de fungicida (estádios R1+R5.1); três (3) aplicações de fungicida (estádios V6+R1+R5.1); quatro (4) aplicações de fungicida (estádios V6+R1+R5.1+R5.4); cinco (5) aplicações de fungicida (estádios V6+R1+R5.1+R5.4+R6); e ausência de aplicação (Testemunha). A severidade foi quantificada pela coleta de trifólios para posterior determinação da Área Abaixo da Curva de Progresso da Doença (AACPD) e do controle (%). Avaliou-se também a produtividade e os componentes de rendimento (número de vagens por planta, número de grãos por vagem e peso de mil sementes). A cv. TMG 7262 expressou sintomas da doença 14 dias após a cv. BMX Tornado, independentemente do número de aplicações de fungicidas. Ambas as cultivares testadas foram responsivas à produtividade com 4 e 5 aplicações de fungicida, enquanto que a correlação entre AACPD e produtividade foi semelhante entre as cultivares