Novel insights into mitochondrial phospholipid homeostasis in a disease-relevant yeast model

The proper function of mitochondria critically depends on their membrane lipid composition. To ensure lipid homeostasis, de novo synthesis, intracellular and intraorganellar transport, remodeling, and degradation of lipids must be tightly regulated. Several studies have emphasised the importance of the mitochondrial signature phospholipid, cardiolipin (CL) for the organelle function. The acquisition of mature CL species is catalyzed by the phospholipid acyltransferase, Tafazzin. The importance of CL remodeling is underscored by the fact that mutations in Tafazzin lead to a life-threatening genetic disorder, Barth syndrome (BTHS). Currently, the biochemical processes underlying this clinical disorder remain unclear. Deletion of the yeast homologue Taz1 results in similar phenotypes to those observed in patients suffering from BTHS, making this organism an optimal model system to study the pathomechanism of the disease. To shed light on the pathomechanism of BTHS, I searched for yeast multi-copy suppressors of the taz1Δ growth defect and identified the branched-chain amino acid transaminases (BCATs) BAT1 and BAT2 as such suppressors. Similarly, overexpression of the mitochondrial isoform BCAT2 in mammalian cells lacking TAZ improves their growth. Accordingly, supplying both yeast and mammalian cells lacking Tafazzin function with certain amino acids restored their growth behavior. Although elevated levels of Bat1 or Bat2 did not restore all the mitochondrial defects of BTHS, it could correct the higher respiration rate observed in taz1Δ cells. These findings outline that the metabolism of amino acids can influence the BTHS phenotype and has an important and disease relevant role in cells lacking Taffazzin function. In another project, I investigated the transfer of lipids between mitochondria and vacuoles. The absence of documented mitochondrial vesicular lipid exchange suggests that membrane contact sites (MCSs) facilitate lipids transport between mitochondria and other cellular membranes. Recently, it has been demonstrated that the lack of one contact site leads to the expansion of an alternative one. Specifically, loss of the ER-mitochondria encounter structure (ERMES) can be bypassed by point mutations in the vacuolar protein Vps13, or by overexpression of the mitochondrial Mdm10 complementing protein 1 (Mcp1). However, the mechanism by which this bypass support lipid homeostasis has remained unclear. In this work, I analyzed the membrane topology of Mcp1. My findings revealed that Mcp1 functions as a recruiter of Vps13 to mitochondria and promotes formation of vacuole-mitochondria MCS. I demonstrated that the N-terminal region of Mcp1 is exposed to the cytosol and mediates the recruitment of Vps13, thus establishing a functional mitochondria-vacuole MCS that compensate for the loss of ERMES. Finally, in a third project of my doctoral studies I investigated the relationship between the ERMES complex and the coenzyme Q6 (CoQ6) biosynthesis system. I observed that supplementation of yeast cells lacking functional ERMES with CoQ6 could rescue the growth retardation and the altered mitochondrial morphology of these mutated cells. Based on additional results from collaborating groups, we suggest that the ERMES complex coordinates coenzyme Q biosynthesis

a importância dos momentos de reflexão na creche e no jardim-de-infância

O presente relatório de estágio tem como tema principal o trabalho em equipa. Aborda com mais pormenor o subtema dos momentos de reflexão na equipa pedagógica. Ao longo do estudo são vários os conceitos abordados e a interligação entre os mesmos. Pretendo dar a conhecer a temática em questão e conceções que lhe estão naturalmente associadas. Sendo um estudo investigativo são abordados os métodos utilizados ao longo de toda a investigação. Com este relatório pretende-se alcançar qual o papel da reflexão numa equipa educativa e, principalmente, pedagógica, compreendendo a importância que tem na prática desenvolvida em contextos de educação de infância. Por fim, faz-se um balanço global sobre as várias vertentes deste relatório investigativo.This report has team work as its main subject. It explores meticulously the reflection moments within the pedagogical team. Along this study multiple concepts and their connections are addressed. My goal is to explore this subject and its natural perceptions. Being a research, this work comprehends the methods used along this investigation. My main goal with this report is to understand the importance of the reflection in an educational team and, most of all, in a pedagogical team, including its practical significance in childhood education context. In conclusion, it was done a global balance concerning the multiple approaches of this investigational report.Escola Superior de Educação, Instituto Politécnico de Setúba

Onychomatricoma on the fourth toenail: a rare tumor in a rare localization

© 2021 Indian Journal of Dermatology, Venereology and Leprology - Published by Scientific Scholar. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-Share Alike 4.0 License.Onychomatricoma is a rare benign tumor of the nail matrix, characterized by finger-like projections that invade the nail plate. The fingernails of Caucasian women are most commonly affected. Because this tumor can easily mimic other more prevalent ungual diseases, it is crucial to be acquainted with its characteristic clinical and histopathologic features. The authors present a case of a 40-year-old man with an onychomatrichoma in the fourth left toenail, which was initially misdiagnosed and treated as onychomycosis.info:eu-repo/semantics/publishedVersio

Lipidómica e proteómica do catabolismo muscular subjacente ao cancro

Mestrado em Bioquímica ClínicaA caquexia associada ao cancro é uma condição fisiopatológica complexa caraterizada por acentuada perda de massa muscular.Recentemente, esta situação foi associada à disfunção mitocondrial. A relação e o papel do proteoma e lipidoma mitocondrial e a funcionalidade deste organelo permanece pouco compreendida, em particular no contexto do catabolismo muscular associado ao cancro. No sentido de melhor compreender os mecanismos moleculares subjacentes às alterações no músculo esquelético na caquexia associada ao cancro, utilizaram-se 23 ratos Wistar divididos aleatoriamente em dois grupos: com cancro da bexiga induzido pela exposição durante 20 semanas a N-butil-N-(4-hidroxibutil)-nitrosamina (grupo BBN, n=13) ou saudáveis (CONT, n=10). No final do protocolo verificou-se que os animais do grupo BBN apresentavam uma perda significativa de peso corporal e de massa muscular. Também foi observado uma diminuição da atividade da fosforilação oxidativa de mitocôndrias isoladas do músculo gastrocnemius. a qual foi acompanhada por alterações do perfil de fosfolípidos (PL) da mitocôndria. A alteração do lipidoma mitocondrial caraterizou-se pelo aumento do teor relativo de fosfatidilcolinas (PC) e fosfatidilserina (PS) e uma redução no teor relativo de cardiolipina (CL), ácido fosfatídico (PA), fosfatidilglicerol (PG) e fosfatidilinositol (PI). A análise realizada por GC-FID e HPLC-ESI-MS evidenciou ainda um aumento de ácidos gordos polinsaturados, com um aumento destacado de C22:6 em PC, PE e PS. A diminuição de CL foi acompanhada por diminuição na expressão de citocromo c e aumento da razão Bax/Bcl2, sugestivo de maior suscetibilidade à apoptose e stress oxidativo. Embora em níveis mais elevados, a UCP-3 não parece proteger as proteínas mitocondriais da lesão oxidativa atendendo ao aumento do teor de proteínas carboniladas. Em conclusão, a remodelação de PL da mitocôndria parece estar associada à disfunção da OXPHOS e, consequentemente, do catabolismo muscular associado ao cancro.Cancer cachexia (CC) is a complex pathophysiological condition characterized by a marked muscle wasting. Recently, this situation has been associated to mitochondrial dysfunction. The interplay and role of mitochondrial proteome and lipidome and also the functionality of this organelle remains poorly understood in the context of cancer-related muscle wasting. To better understand the molecular mechanisms underlying skeletal muscle wasting, 23 Wistar rats were randomly divided in two groups: animals with bladder cancer induced by the exposition to N-butyl-N-(4-hydroxybutyl)-nitrosamine for 20 weeks (BBN, n=13) or healthy ones (CONT, n=10). At the end of the experimental protocol, BBN animals demonstrated a significant body weight and muscle mass loss and was also observed an decreased activity of oxidative phosphorylation in mitochondria isolated from gastrocnemius muscle, which was accompanied by alterations of this organelle’s phospholipids (PL) profile. The mitochondrial lipidome alterations were characterized by an increase of the relative content of phosphatidylcholines (PC) and phosphatidylserine (PS) and a decrease of cardiolipin (CL), phosphatidic acid (PA), phosphatidylglycerol (PG) and phosphatidylinositol (PI). GC-FID and HPLC-ESI-MS analysis also showed an increase of polyunsaturated fatty acids, particularly of C22:6 in PC, PE and PS. The observed decrease in CL class was accompanied by a decrease in the expression of cytochrome c, and an increase of the ratio Bax/Bcl-2, suggestive of a greater susceptibility to apoptosis and oxidative stress. Although in higher levels, UCP-3 does not seem to protect mitochondrial proteins from oxidative damage considering the increased content of carbonylated protein. In conclusion, the PL remodeling seems to be associated to OXPHOS dysfunction and consequently to muscle catabolism associated with cancer

Intraspecific diversity of A. tetracladia affects plant-litter decomposition in freshwaters

FEDER-POFC-COMPETE and FCT supported this study (PEst-C/BIA/UI4050/2011, PTDC/AAC-AMB/113746/2009) and SD (SFRH/BPD/47574/2008).Fundação para a Ciência e a Tecnologia (FCT) - PEst-C/BIA/UI4050/2011, PTDC/AAC-AMB/113746/2009, SFRH/BPD/47574/200

Duas Novas Mutações do Gene ATP2C1 em Doentes Portuguesas com Doença de Hailey-Hailey

Hailey-Hailey disease (HHD) is a rare autosomal dominant acantholytic dermatosis. It is characterized by a recurrent eruption of vesicles, erosions, and scaly erythematous plaques involving intertriginous areas and first occurring after puberty, mostly in the third or fourth decade. In 2000, mutations in the ATP2C1 gene on band 3q22.1, encoding the secretory pathway Ca2+/Mn2+-ATPase protein 1(hSPCA1), have been identified as the cause of HHD. We report the identification of two novel mutations of ATP2C1 gene in two Portuguese patients, which expands the spectrum of ATP2C1 mutations underlying HHD and provides useful information for genetic counseling.A doença de Hailey-Hailey (DHH) é uma dermatose acantolítica autossómica dominante rara. Clinicamente, caracteriza-se por episódios recorrentes de vesículas, erosões e placas eritematosas descamativas envolvendo áreas intertriginosas, com início após a puberdade, geralmente na terceira ou quarta década de vida. Em 2000, mutações no gene ATP2C1, no cromossoma 3q22.1, que codifica a proteína 1 da via secretora humana Ca2+/Mn2+-ATPase (hSPCA1), foram identificadas como a causa da DHH. Relatamos a identificação de duas novas mutações do gene ATP2C1 em duas doentes portuguesas, expandido o espectro de mutações ATP2C1 subjacentes à DHH e fornecendo informações úteis para o aconselhamento genético

Desenho do Mapa da Estratégia da Área Funcional de Saúde Ambiental da Unidade de Saúde Pública Amélia Leitão em Cascais (Portugal)

This paper presents the road traveled to draw the strategic map of the Public Health Unit’s Environmental Health Functional Area associated with the geographical area of the municipality of Cascais in Portugal. The applied methodology was the Balanced Scorecard management model, its four dimensions being taken into account: customers, processes, learning and finance. In addition to the Strategic Map, SWOT and stakeholder analyses were carried out, and the vision, the mission and the values of the organization, among others, were defined. It is important to make progress toward a consolidated map, the immediate priorities being to identify key processes and to establish procedures therefor.Este documento presenta el camino recorrido en la elaboración del mapa estratégico del área funcional de salud ambiental de la unidad de salud pública, relativa al área geográfica del municipio de Cascais en Portugal. La metodología adoptada fue la del modelo de gestión del Cuadro de Mando Integral considerando sus cuatro dimensiones: clientes, procesos, aprendizaje y financiero. Además del Mapa Estratégico, se elaboraron, necesariamente, análisis DAFO y de stakeholders y se definieron la visión, misión y valores de la organización, entre otros. Es importante avanzar hacia un mapa consolidado destacando como prioridad inmediata, la identificación de los procesos clave y la definición de procedimientos para los mismos.Neste documento descreve-se o percurso efetuado na elaboração do mapa da estratégia da área funcional de saúde ambiental da unidade de saúde pública, com influência na área geográfica do concelho de Cascais em Portugal. A metodologia adotada foi a do modelo de gestão Balanced Scorecard considerando as suas quatro dimensões: clientes, processos, aprendizagem e financeira. Para além do Mapa da Estratégia, necessariamente, foram elaboradas as análises de stakeholders e SWOT e definidas a visão, missão e valores da organização, entre outros. Importa caminhar para um mapa consolidado destacando-se no imediato como prioritária a identificação dos processos-chave e a definição de procedimentos para os mesmos

Enterococcus spp. from chicken meat collected 20 years apart overcome multiple stresses occurring in the poultry production chain : Antibiotics, copper and acids

Poultry meat has been a vehicle of antibiotic resistant bacteria and genes. Yet, the diversity of selective pressures associated with their maintenance in the poultry-production chain remains poorly explored. We evaluated the susceptibility of Enterococcus spp. from chicken meat collected 20 years apart to antibiotics, metals, acidic pH and peracetic acid-PAA. Contemporary chicken-meat samples (n = 53 batches, each including a pool of neck skin from 10 single carcasses) were collected in a slaughterhouse facility using PAA as disinfectant (March-August 2018, North of Portugal). Broilers were raised in intensive farms (n = 29) using CuSO4 and organic acids as feed additives. Data were compared with that of 67 samples recovered in the same region during 1999-2001. All 2018 samples had multidrug resistant-MDR isolates, with >45 % carrying Enterococcus faecalis, Enterococcus faecium or Enterococcus gallinarum resistant to tetracycline, erythromycin, ampicillin, quinupristin-dalfopristin, ciprofloxa-cin, chloramphenicol or aminoglycosides. Resistance rates were similar (P > 0.05) to those of 1999-2001 samples for all but five antibiotics. The decrease of samples carrying vancomycin-resistant isolates from 46 % to % between 1999-2001 and 2018 was the most striking difference. Isolates from both periods were similarly susceptible to acid pH [minimum-growth pH (4.5-5.0), minimum-survival pH (3.0-4.0)] and to PAA (MIC90 = 100-120 mg/L/MBC90 = 140-160 mg/L; below concentrations used in slaughterhouse). Copper tolerance genes (tcrB and/or cueO) were respectively detected in 21 % and 4 % of 2018 and 1999-2001 samples. The tcrB gene was only detected in E. faecalis (MICCuSO4 > 12 mM), and their genomes were compared with other international ones of chicken origin (PATRIC database), revealing a polyclonal population and a plasmid or chromosomal location for tcrB. The tcrB plasmids shared diverse genetic modules, including multiple antimicrobial resistance genes (e.g. to tetracyclines, chloramphenicol, macrolide-lincosamide-streptogramin B-MLSB, aminoglycosides, bacitracin, coccidiostats). When in chromosome, the tcrB gene was co-located closely to merA (mercury) genes. Chicken meat remains an important vehicle of MDR Enterococcus spp. able to survive under diverse stresses (e.g. copper, acid) potentially contributing to these bacteria maintenance and flux among animal-environment -humans.Peer reviewe

Treating Parkinson's Disease with Human Bone Marrow Mesenchymal Stem Cell Secretome: A Translational Investigation Using Human Brain Organoids and Different Routes of In Vivo Administration.

peer reviewedParkinson's disease (PD) is the most common movement disorder, characterized by the progressive loss of dopaminergic neurons from the nigrostriatal system. Currently, there is no treatment that retards disease progression or reverses damage prior to the time of clinical diagnosis. Mesenchymal stem cells (MSCs) are one of the most extensively studied cell sources for regenerative medicine applications, particularly due to the release of soluble factors and vesicles, known as secretome. The main goal of this work was to address the therapeutic potential of the secretome collected from bone-marrow-derived MSCs (BM-MSCs) using different models of the disease. Firstly, we took advantage of an optimized human midbrain-specific organoid system to model PD in vitro using a neurotoxin-induced model through 6-hydroxydopamine (6-OHDA) exposure. In vivo, we evaluated the effects of BM-MSC secretome comparing two different routes of secretome administration: intracerebral injections (a two-site single administration) against multiple systemic administration. The secretome of BM-MSCs was able to protect from dopaminergic neuronal loss, these effects being more evident in vivo. The BM-MSC secretome led to motor function recovery and dopaminergic loss protection; however, multiple systemic administrations resulted in larger therapeutic effects, making this result extremely relevant for potential future clinical applications

Mycobacterium leprae diversity and population dynamics in medieval Europe from novel ancient genomes.

Funder: Max-Planck SocietyFunder: St John’s College, CambridgeFunder: Fondation Raoul FollereauFunder: University of Zurich’s University Research Priority Program “Evolution in Action: From Genomes to Ecosystems”Funder: the Senckenberg Centre for Human Evolution and Palaeoenvironment (S-HEP) at the University of TübingenBackgroundHansen's disease (leprosy), widespread in medieval Europe, is today mainly prevalent in tropical and subtropical regions with around 200,000 new cases reported annually. Despite its long history and appearance in historical records, its origins and past dissemination patterns are still widely unknown. Applying ancient DNA approaches to its major causative agent, Mycobacterium leprae, can significantly improve our understanding of the disease's complex history. Previous studies have identified a high genetic continuity of the pathogen over the last 1500 years and the existence of at least four M. leprae lineages in some parts of Europe since the Early Medieval period.ResultsHere, we reconstructed 19 ancient M. leprae genomes to further investigate M. leprae's genetic variation in Europe, with a dedicated focus on bacterial genomes from previously unstudied regions (Belarus, Iberia, Russia, Scotland), from multiple sites in a single region (Cambridgeshire, England), and from two Iberian leprosaria. Overall, our data confirm the existence of similar phylogeographic patterns across Europe, including high diversity in leprosaria. Further, we identified a new genotype in Belarus. By doubling the number of complete ancient M. leprae genomes, our results improve our knowledge of the past phylogeography of M. leprae and reveal a particularly high M. leprae diversity in European medieval leprosaria.ConclusionsOur findings allow us to detect similar patterns of strain diversity across Europe with branch 3 as the most common branch and the leprosaria as centers for high diversity. The higher resolution of our phylogeny tree also refined our understanding of the interspecies transfer between red squirrels and humans pointing to a late antique/early medieval transmission. Furthermore, with our new estimates on the past population diversity of M. leprae, we gained first insights into the disease's global history in relation to major historic events such as the Roman expansion or the beginning of the regular transatlantic long distance trade. In summary, our findings highlight how studying ancient M. leprae genomes worldwide improves our understanding of leprosy's global history and can contribute to current models of M. leprae's worldwide dissemination, including interspecies transmissions