Análise comparativa entre isolados de Fonsecaea sp na modulação da resposta imune do hospedeiro : papel das vesículas e componentes extracelulares

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Instituto de Ciências Biológicas, Departamento de Biologia Molecular, 2021.A cromoblastomicose (CBM) é uma micose cutânea e subcutânea crônica, causada por fungos negros, dimórficos e filamentosos da família Dematiaceae. Fonsecaea pedrosoi é considerado uma das principais espécies causadoras da doença no Brasil, apesar de que F. monophora, F. nubica e F. pugnacius terem sido descritas como outras espécies capazes de causar CBM em humanos. Todas estas espécies possuem a capacidade de transitar entre as formas saprofíticas, conídios e hifas, e a forma patogênica do fungo, as células muriformes (MCs). Fatores de virulência produzidos por fungos, como vesículas extracelulares (EVs) e moléculas extracelulares presentes no meio condicionado (CM) vem sendo mais estudados nos últimos anos, apesar de existirem poucas informações sobre o papel destas moléculas extracelulares durante a CBM. Este trabalho visa compreender os padrões imunoestimulatórios in vitro e in vivo causados por EVs e CMs produzidos por F. pedrosoi, F. monophora, F. nubica e F. erecta isolados a partir de conídios e hifas cultivados em meio rico (RM) e meio mínimo (MM), além de analisar o papel dessas moléculas quando produzidas na presença ou ausência de MCs. Em relação as diferentes condições nutricionais de cultivo, foi demonstrado que EVs produzidas em RM tem maior capacidade de estimular a produção de TNF, IL-1β e IL-10 por BMDMs em comparação a EVs produzidas em MM na presença ou na ausência de MCs. Tais resultados são diferentes dos observados para EVs de C. neoformans produzidas em RM e MM, em que EVs isoladas de MM demonstram estimular mais a produção de citocinas pró-inflamatórias do que em RM. Já os CMs demonstram ser mais pró-inflamatórios do que as EVs, principalmente os isolados a partir de MM na ausência de MCs, aumentando os níveis de secreção das citocinas TNF, IL-1β e IL-10 in vitro. Por fim, os CMs produzidos por F. pedrosoi e F. erecta foram utilizados como tratamento in vivo e demonstraram agravar a progressão da CBM em modelo murino, aumentando o diâmetro da área da lesão, o número de unidades formadoras de colônia e os níveis das citocinas pró-inflamatórias TNF e IL-1β em relação ao controle positivo tratado com PBS. Já foi observado que as moléculas presentes no CM produzidos pela forma capsular de C. neoformans conseguem inibir a ativação do inflamassoma NLRP3 e promover a proliferação celular in vitro, indicando que as moléculas presentes nos CMs de fungos patogênicos possuem potencial de imunomodular a resposta imune do hospedeiro. Desta forma, observa-se que as EVs e os CMs produzidos por diferentes espécies do gênero Fonsecaea possuem a capacidade tanto de estimular, quanto de inibir a secreção de citocinas importantes na resposta imune inata como TNF, IL-1β e IL-10, demonstrando serem moléculas com capacidade de virulência e possivelmente, de modular a progressão da CBM em humanos.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous mycosis caused by black, dimorphic and filamentous fungi of the Dematiaceae family. Fonsecaea pedrosoi is considered one of the main species causing the disease, although F. monophora, F. nubica and F. pugnacious are considered as other species capable of causing CBM in humans. All these species have an ability to transition between saprophytic forms, such as conidia and hyphae and the pathogenic form of the fungus, the murine cells (MCs). Virulence factors produced by fungi, such as extracellular vesicles (EVs) and conditioned media (CM) have been more studied in recent years, although there is little information about the roles of these extracellular molecules during CBM. This work aims to understand the in vitro and in vivo immunostimulatory patterns caused by EVs and CMs obtained by F. pedrosoi, F. monophora, F. nubica and F. erecta isolated from conidia and hyphae cultivated in rich medium (RM) and minimal medium (MM), in addition to analyzing the role of these molecules when produced in the presence or absence of MCs. Regarding the different nutritional conditions of cultivation, it was shown that EVs produced in RM have a greater capacity to stimulate the production of TNF, IL-1β and IL-10 by BMDMs when compared to EVs produced in MM in the presence or absence of MCs. These results are different from those observed for EVs of C. neoformans produced in RM and MM, in which EVs isolated from MM are shown to stimulate more the production of pro-inflammatory cytokines than in RM. The CMs, on the other hand, demonstrate to be more pro-inflammatory than EVs, especially those isolated from MM in the absence of MCs, which increased the levels of cytokine secretion of TNF, IL-1β and IL-10 in vitro. Finally, the CMs produced from F. pedrosoi and F. erecta were used as an in vivo treatment and where shown to aggravate the progression of CBM in a murine model, increasing the diameter of the lesion area, the number of colony forming units and the levels of pro-inflammatory cytokines such as TNF and IL-1β, compared to the positive control treated with PBS. The role of the molecules presented in the CM produced by capsular C. neoformans has already been observed to inhibit inflamassome NLRP3 activation and promote cell proliferation in vitro, indicating that theses extracellular molecules produced by pathogenic fungi have the potential to immunomodulate the host’s immune response. Thus, it is observed that the EVs produced by different species of the Fonsecaea genus have the ability to both stimulate and inhibit important cytokines in the innate immune response, proving to be molecules with high virulence capacity and possible mediators of the CBM progression in humans

Chemical modeling for pH prediction of acidified musts with gypsum and tartaric acid in warm regions

Winemaking of musts acidified with up to 3 g/L of gypsum (CaSO4 2H2O) and tartaric acid, both individually and in combination, as well as a chemical modeling have been carried out to study the behaviour of these compounds as acidifiers. Prior to fermentation gypsum and tartaric acid reduce the pH by 0.12 and 0.17 pH units/g/L, respectively, but while gypsum does not increase the total acidity and reduces buffering power, tartaric acid shows the opposite behaviour. When these compounds were used in combination, the doses of tartaric acid necessary to reach a suitable pH were reduced. Calcium concentrations increase considerably in gypsum-acidified must, although they fell markedly after fermentation over time. Sulfate concentrations also increased, although with doses of 2 g/L they were lower than the maximum permitted level (2.5 g/L). Chemical modeling gave good results and the errors in pH predictions were less than 5% in almost all case

Searching for realistic 4d string models with a Pati-Salam symmetry -- Orbifold grand unified theories from heterotic string compactification on a Z6 orbifold

Motivated by orbifold grand unified theories, we construct a class of three-family Pati-Salam models in a Z6 abelian symmetric orbifold with two discrete Wilson lines. These models have marked differences from previously-constructed three-family models in prime-order orbifolds. In the limit where one of the six compactified dimensions (which lies in a Z2 sub-orbifold) is large compared to the string length scale, our models reproduce the supersymmetry and gauge symmetry breaking pattern of 5d orbifold grand unified theories on an S1/Z2 orbicircle. We find a horizontal 2+1 splitting in the chiral matter spectra -- 2 families of matter are localized on the Z2 orbifold fixed points, and 1 family propagates in the 5d bulk -- and identify them as the first-two and third families. Remarkably, the first two families enjoy a non-abelian dihedral D4 family symmetry, due to the geometric setup of the compactified space. In all our models there are always some color triplets, i.e. (6,1,1) representations of the Pati-Salam group, survive orbifold projections. They could be utilized to spontaneously break the Pati-Salam symmetry to that of the Standard Model. One model, with a 5d E6 symmetry, may give rise to interesting low energy phenomenology. We study gauge coupling unification, allowed Yukawa couplings and some of their phenomenological consequences. The E6 model has a renormalizable Yukawa coupling only for the third family. It predicts a gauge-Yukawa unification relation at the 5d compactification scale, and is capable of generating reasonable quark/lepton masses and mixings. Potential problems are also addressed, they may point to the direction for refining our models.Comment: 58 pages, 5 figures, 4 tables, revtex4 with ams fonts. Version to appear in NP

Does Accessibility Planning address what matters? A review of current practice and practitioner perspectives

'Accessibility' has become commonplace in transport planning and as such there is a plethora of interpretations of what accessibility means, what constitutes a good measure of accessibility, and how this might be applied in practice. This paper presents an overview of approaches to measuring accessibility and presents a case study of Accessibility Planning in England — one approach to formalising the concept of accessibility. Results of semi-structured interviews with local authority officers are discussed to establish whether current approaches, allow their desired outcomes to be met. This approach demonstrates where there might be gaps between measured or modelled accessibility and the perceptions of the individuals. Findings suggest that while the process is deemed useful in raising the profile of accessibility issues, measures of accessibility do not necessarily easily translate into quantifying benefits of those improvements that are perceived by practitioners to improve accessibility and reduce transport disadvantage

Comportamiento agronómico de cinco variedades de trigo en el centro de la Provincia de Córdoba

Trabajo Final Integrador (Área de Consolidación Sistemas Agrícolas de Producción Extensivos - Ingeniería Agronómica) -- UNC- Facultad de Ciencias Agropecuarias, 2018La importancia del cultivo de trigo radica en el impacto que éste genera sobre el sistema productivo, entre los que se pueden mencionar: aportes al margen bruto de las empresas, aporte financiero al inicio de la campaña gruesa, aporte de diversidad, aportes a la siembra directa (distribución y calidad de los rastrojos, mejora en la retención e infiltración del agua, disminución de las perdidas por evaporación, prevención de la erosión eólica e hídrica), alta eficiencia del uso del agua almacenada en el suelo y junto con el maíz son los que aportan mayor beneficio al balance del carbono. El objetivo de este trabajo fue analizar el comportamiento de 5 variedades de trigo en el centro de la provincia de Córdoba. El ensayo se realizó en el Campo Escuela de la Facultad de Ciencias Agropecuarias de la U.N.C. Cada unidad experimental tuvo una dimensión de 87,5 m 2 , conformado por 25 surcos (4,37 metros de ancho) y un largo de 20 metros. No hubo repeticiones de las unidades experimentales. Las variables analizadas fueron rendimiento, sus componentes, aptitud panadera y el perfil sanitario de las cinco variedades. Los rendimientos obtenidos fueron bajos si consideramos al agua y al nitrógeno disponible. Todas las variedades tuvieron un bajo peso hectolitrico y peso de los 1000 granos. A excepción de una, mostraron una red de gluten fuerte y tenaz siendo apta para panificación. Drechslera tritici-repentis y Puccinia striiformis fueron las que más daño causaron, siendo la variedad V 16 la más afectada por esta

Cambios en el perfil bioquímico hepático de alpacas positivas a huevos de Fasciola hepática

The aim of this study was to determine changes in the liver biochemical profile in alpacas with presence of F. hepatica eggs. We used serum and feces samples of 35 male alpaca between 2-5 years old, 18 were positives and 17 were negatives in coproparasitologic analysis for F. hepatica eggs. Differences are showed (p<0.05) in the values of ALT, GGT and direct bilirubin between positives y negatives animals. We concluded that there is relationship between presence of F. hepatica eggs and high levels in the liver biochemical profile, maybe due to alterations in the hepatic parenchyma by chronic infection in alpacas.El objetivo del presente estudio fue determinar cambios en el perfil bioquímico hepático en alpacas con presencia de huevos de F. hepatica en heces. Se utilizaron muestras de suero y heces de 35 alpacas machos de entre 2 y 5 años de edad, de los cuales 18 fueron positivos y 17 negativos al examen coproparasitológico (sedimentación) para identificación de huevos de F. hepática. Se observó diferencia significativa (p<0.05) entre los valores de ALT, GGT y bilirrubina directa entre los animales positivos y negativos. Concluimos que la presencia de huevos de Fasciola hepatica guarda relación con la elevación de algunos parámetros del perfil hepática posiblemente por alguna alteración en el parénquima producto de una infección crónica en alpacas

Teaching and learning musical instruments through ICT: the impact of the COVID-19 pandemic lockdown

The COVID-19 lockdown in education institutions required music teachers to use ICTto continue teaching. This research study, with the use of a Likert type online questionnaire, analyses the ICT activities carried out during this period and the learning conceptions they reflect. The questionnaire consisted of the description of activities which varied, depending on the learning promoted (reproductive or constructive), the learning outcomes (verbal, procedural, or attitudinal), the type of assessment to which the activities were directed, and the presence of cooperative activities. The teachers had to indicate the frequency with which they carried out these activities. The questionnaire was completed by 254 instrumental music teachers from different types of institutions and different levels. The main study outcome was that teachers used reproductive activities more frequently than constructive ones. We also found that most activities were those favouring verbal learning and assessment. The cooperative activities were the least frequent. Finally, through a cluster analysis, we identified three teaching profiles depending on the frequency and type of ICT used: Passive, Active, and Interpretative. The variable that produced the most consistent differences was previous ICT useThis work was supported by Ministerio de Ciencia e Innovacion of Spain (PID2020-114177RB-I00). Guadalupe Lopez- Iñiguez was also supported by the Senior Researcher grant “Expanding reflexivity of professional education in music in times of crises” awarded by the Jenny and Antti Wihuri Foundation in Finlan

Integrated miRNA/mRNA Counter-Expression Analysis Highlights Oxidative Stress-Related Genes CCR7 and FOXO1 as Blood Markers of Coronary Arterial Disease

Our interest in the mechanisms of atherosclerosis progression (ATHp) has led to the recent identification of 13 miRNAs and 1285 mRNAs whose expression was altered during ATHp. Here, we deepen the functional relationship among these 13 miRNAs and genes associated to oxidative stress, a crucial step in the onset and progression of vascular disease. We first compiled a list of genes associated to the response to oxidative stress (Oxstress genes) by performing a reverse Gene Ontology analysis (rGO, from the GO terms to the genes) with the GO terms GO0006979, GO1902882, GO1902883 and GO1902884, which included a total of 417 unique Oxstress genes. Next, we identified 108 putative targets of the 13 miRNAs among these unique Oxstress genes, which were validated by an integrated miRNA/mRNA counter-expression analysis with the 1285 mRNAs that yielded 14 genes, Map2k1, Mapk1, Mapk9, Dapk1, Atp2a2, Gata4, Fos, Egfr, Foxo1, Ccr7, Vkorc1l1, Rnf7, Kcnh3, and Mgat3. GO enrichment analysis and a protein–protein-interaction network analysis (PPI) identified most of the validated Oxstress transcripts as components of signaling pathways, highlighting a role for MAP signaling in ATHp. Lastly, expression of these Oxstress transcripts was measured in PBMCs from patients suffering severe coronary artery disease, a serious consequence of ATHp. This allowed the identification of FOXO1 and CCR7 as blood markers downregulated in CAD. These results are discussed in the context of the interaction of the Oxstress transcripts with the ATHp-associated miRNAs

Chronic kidney disease-associated inflammation increases in risks of acute kidney injury and mortality after cardíac surgery

Cardiovascular mortality increases with decreasing renal function although the cause is yet unknown. Here, we have investigated whether low chronic inflammation in chronic kidney diseases (CKD) could contribute to increased risk for coronary artery diseases (CAD). Thus, a prospective case-control study was conducted in patients with CAD and CKD undergoing coronary artery bypass graft surgery with the aim of detecting differences in cardiovascular outcomes, epicardial adipose tissue volume, and inflammatory marker activity associated with renal dysfunction. Expression of membrane CD14 and CD16, inflammatory cytokines and chemokines, mitogen-activated protein (MAP) kinases and hsa-miR-30a-5p were analyzed in peripheral blood mononuclear cells (PBMCs). Epicardial fat volume and tissue inflammation in perivascular adipose tissue and in the aorta were also studied. In the present study, 151 patients were included, 110 with CAD (51 with CKD) and 41 nonCAD controls (15 with CKD). CKD increased the risk of cardiac surgery-associated acute kidney injury (CSA-AKI) as well as the 30-day mortality after cardiac surgery. Higher counts of CD14++CD16+ monocytes were associated with vascular inflammation, with an increased expression of IL1β, and with CKD in CAD patients. Expression of hsa-miR-30a-5p was correlated with hypertension. We conclude that CKD patients show an increased risk of CSA-AKI and mortality after cardiovascular surgery, associated with the expansion of the CD14++CD16+ subset of proinflammatory monocytes and with IL1β expression. We propose that inflammation associated with CKD may contribute to atherosclerosis (ATH) pathogenesis

Sensibilidad de la microestructura en un sistema multi-escala

Este trabajo analiza la sensibilidad de los parámetros de optimización de una microestructura periódica isotrópica en la optimización topológica de una viga MBB. La función objetivo para optimización de la microestructura es el módulo volumétrico, y los parámetros analizados son: fracción volumétrica, radio de filtro y factor de penalización. En la macroestructura la función objetivo es la minimización de la flexibilidad, sujeta a un volumen predefinido de 0.5. Se desarrolló un algoritmo, con base en el método SIMP, capaz de transferir las informaciones de la microestructura para la macroestructura. Se realizaron análisis numéricos, variando los parámetros y observando el comportamiento de las soluciones. Los resultados demuestran que la fracción volumétrica de la microestructura tiene gran influencia en la maximización de la rigidez, sin embargo el radio de filtro y el factor de penalización presentaron poca influencia, así como sobre el perfil de la macroestructura.Publicado en: Mecánica Computacional vol. XXXV, no. 23Facultad de Ingenierí