9,631 research outputs found

    Pollution-induced community tolerance in freshwater biofilms – from molecular mechanisms to loss of community functions

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    Exposure to herbicides poses a threat to aquatic biofilms by affecting their community structure, physiology and function. These changes render biofilms to become more tolerant, but on the downside community tolerance has ecologic costs. A concept that addresses induced community tolerance to a pollutant (PICT) was introduced by Blanck and Wängberg (1988). The basic principle of the concept is that microbial communities undergo pollution-induced succession when exposed to a pollutant over a long period of time, which changes communities structurally and functionally and enhancing tolerance to the pollutant exposure. However, the mechanisms of tolerance and the ecologic consequences were hardly studied up to date. This thesis addresses the structural and functional changes in biofilm communities and applies modern molecular methods to unravel molecular tolerance mechanisms. Two different freshwater biofilm communities were cultivated for a period of five weeks, with one of the communities being contaminated with 4 μg L-1 diuron. Subsequently, the communities were characterized for structural and functional differences, especially focusing on their crucial role of photosynthesis. The community structure of the autotrophs was assessed using HPLC-based pigment analysis and their functional alterations were investigated using Imaging-PAM fluorometry to study photosynthesis and community oxygen profiling to determine net primary production. Then, the molecular fingerprints of the communities were measured with meta-transcriptomics (RNA-Seq) and GC-based community metabolomics approaches and analyzed with respect to changes in their molecular functions. The communities were acute exposed to diuron for one hour in a dose-response design, to reveal a potential PICT and uncover related adaptation to diuron exposure. The combination of apical and molecular methods in a dose-response design enabled the linkage of functional effects of diuron exposure and underlying molecular mechanisms based on a sensitivity analysis. Chronic exposure to diuron impaired freshwater biofilms in their biomass accrual. The contaminated communities particularly lost autotrophic biomass, reflected by the decrease in specific chlorophyll a content. This loss was associated with a change in the molecular fingerprint of the communities, which substantiates structural and physiological changes. The decline in autotrophic biomass could be due to a primary loss of sensitive autotrophic organisms caused by the selection of better adapted species in the course of chronic exposure. Related to this hypothesis, an increase in diuron tolerance has been detected in the contaminated communities and molecular mechanisms facilitating tolerance have been found. It was shown that genes of the photosystem, reductive-pentose phosphate cycle and arginine metabolism were differentially expressed among the communities and that an increased amount of potential antioxidant degradation products was found in the contaminated communities. This led to the hypothesis that contaminated communities may have adapted to oxidative stress, making them less sensitive to diuron exposure. Moreover, the photosynthetic light harvesting complex was altered and the photoprotective xanthophyll cycle was increased in the contaminated communities. Despite these adaptation strategies, the loss of autotrophic biomass has been shown to impair primary production. This impairment persisted even under repeated short-term exposure, so that the tolerance mechanisms cannot safeguard primary production as a key function in aquatic systems.:1. The effect of chemicals on organisms and their functions .............................. 1 1.1 Welcome to the anthropocene .......................................................................... 1 1.2 From cellular stress responses to ecosystem resilience ................................... 3 1.2.1 The individual pursuit for homeostasis ....................................................... 3 1.2.2 Stability from diversity ................................................................................. 5 1.3 Community ecotoxicology - a step forward in monitoring the effects of chemical pollution? ................................................................................................................. 6 1.4 Functional ecotoxicological assessment of microbial communities ................... 9 1.5 Molecular tools – the key to a mechanistic understanding of stressor effects from a functional perspective in microbial communities? ...................................... 12 2. Aims and Hypothesis ......................................................................................... 14 2.1 Research question .......................................................................................... 14 2.2 Hypothesis and outline .................................................................................... 15 2.3 Experimental approach & concept .................................................................. 16 2.3.1 Aquatic freshwater biofilms as model community ..................................... 16 2.3.2 Diuron as model herbicide ........................................................................ 17 2.3.3 Experimental design ................................................................................. 18 3. Structural and physiological changes in microbial communities after chronic exposure - PICT and altered functional capacity ................................................. 21 3.1 Introduction ..................................................................................................... 21 3.2 Methods .......................................................................................................... 23 3.2.1 Biofilm cultivation ...................................................................................... 23 3.2.2 Dry weight and autotrophic index ............................................................. 23 3.2.4 Pigment analysis of periphyton ................................................................. 23 3.2.4.1 In-vivo pigment analysis for community characterization ....................... 24 3.2.4.2 In-vivo pigment analysis based on Imaging-PAM fluorometry ............... 24 3.2.4.3 In-vivo pigment fluorescence for tolerance detection ............................. 26 3.2.4.4 Ex-vivo pigment analysis by high-pressure liquid-chromatography ....... 27 3.2.5 Community oxygen metabolism measurements ....................................... 28 3.3 Results and discussion ................................................................................... 29 3.3.1 Comparison of the structural community parameters ............................... 29 3.3.2 Photosynthetic activity and primary production of the communities after selection phase ................................................................................................. 33 3.3.3 Acquisition of photosynthetic tolerance .................................................... 34 3.3.4 Primary production at exposure conditions ............................................... 36 3.3.5 Tolerance detection in primary production ................................................ 37 3.4 Summary and Conclusion ........................................................................... 40 4. Community gene expression analysis by meta-transcriptomics ................... 41 4.1 Introduction to meta-transcriptomics ............................................................... 41 4.2. Methods ......................................................................................................... 43 4.2.1 Sampling and RNA extraction................................................................... 43 4.2.2 RNA sequencing analysis ......................................................................... 44 4.2.3 Data assembly and processing................................................................. 45 4.2.4 Prioritization of contigs and annotation ..................................................... 47 4.2.5 Sensitivity analysis of biological processes .............................................. 48 4.3 Results and discussion ................................................................................... 48 4.3.1 Characterization of the meta-transcriptomic fingerprints .......................... 49 4.3.2 Insights into community stress response mechanisms using trend analysis (DRomic’s) ......................................................................................................... 51 4.3.3 Response pattern in the isoform PS genes .............................................. 63 4.5 Summary and conclusion ................................................................................ 65 5. Community metabolome analysis ..................................................................... 66 5.1 Introduction to community metabolomics ........................................................ 66 5.2 Methods .......................................................................................................... 68 5.2.1 Sampling, metabolite extraction and derivatisation................................... 68 5.2.2 GC-TOF-MS analysis ............................................................................... 69 5.2.3 Data processing and statistical analysis ................................................... 69 5.3 Results and discussion ................................................................................... 70 5.3.1 Characterization of the metabolic fingerprints .......................................... 70 5.3.2 Difference in the metabolic fingerprints .................................................... 71 5.3.3 Differential metabolic responses of the communities to short-term exposure of diuron ............................................................................................................ 73 5.4 Summary and conclusion ................................................................................ 78 6. Synthesis ............................................................................................................. 79 6.1 Approaches and challenges for linking molecular data to functional measurements ...................................................................................................... 79 6.2 Methods .......................................................................................................... 83 6.2.1 Summary on the data ............................................................................... 83 6.2.2 Aggregation of molecular data to index values (TELI and MELI) .............. 83 6.2.3 Functional annotation of contigs and metabolites using KEGG ................ 83 6.3 Results and discussion ................................................................................... 85 6.3.1 Results of aggregation techniques ........................................................... 85 6.3.2 Sensitivity analysis of the different molecular approaches and endpoints 86 6.3.3 Mechanistic view of the molecular stress responses based on KEGG functions ............................................................................................................ 89 6.4 Consolidation of the results – holistic interpretation and discussion ............... 93 6.4.1 Adaptation to chronic diuron exposure - from molecular changes to community effects.............................................................................................. 93 6.4.2 Assessment of the ecological costs of Pollution-induced community tolerance based on primary production ............................................................. 94 6.5 Outlook ............................................................................................................ 9

    Estudo da remodelagem reversa miocárdica através da análise proteómica do miocárdio e do líquido pericárdico

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    Valve replacement remains as the standard therapeutic option for aortic stenosis patients, aiming at abolishing pressure overload and triggering myocardial reverse remodeling. However, despite the instant hemodynamic benefit, not all patients show complete regression of myocardial hypertrophy, being at higher risk for adverse outcomes, such as heart failure. The current comprehension of the biological mechanisms underlying an incomplete reverse remodeling is far from complete. Furthermore, definitive prognostic tools and ancillary therapies to improve the outcome of the patients undergoing valve replacement are missing. To help abridge these gaps, a combined myocardial (phospho)proteomics and pericardial fluid proteomics approach was followed, taking advantage of human biopsies and pericardial fluid collected during surgery and whose origin anticipated a wealth of molecular information contained therein. From over 1800 and 750 proteins identified, respectively, in the myocardium and in the pericardial fluid of aortic stenosis patients, a total of 90 dysregulated proteins were detected. Gene annotation and pathway enrichment analyses, together with discriminant analysis, are compatible with a scenario of increased pro-hypertrophic gene expression and protein synthesis, defective ubiquitinproteasome system activity, proclivity to cell death (potentially fed by complement activity and other extrinsic factors, such as death receptor activators), acute-phase response, immune system activation and fibrosis. Specific validation of some targets through immunoblot techniques and correlation with clinical data pointed to complement C3 β chain, Muscle Ring Finger protein 1 (MuRF1) and the dual-specificity Tyr-phosphorylation regulated kinase 1A (DYRK1A) as potential markers of an incomplete response. In addition, kinase prediction from phosphoproteome data suggests that the modulation of casein kinase 2, the family of IκB kinases, glycogen synthase kinase 3 and DYRK1A may help improve the outcome of patients undergoing valve replacement. Particularly, functional studies with DYRK1A+/- cardiomyocytes show that this kinase may be an important target to treat cardiac dysfunction, provided that mutant cells presented a different response to stretch and reduced ability to develop force (active tension). This study opens many avenues in post-aortic valve replacement reverse remodeling research. In the future, gain-of-function and/or loss-of-function studies with isolated cardiomyocytes or with animal models of aortic bandingdebanding will help disclose the efficacy of targeting the surrogate therapeutic targets. Besides, clinical studies in larger cohorts will bring definitive proof of complement C3, MuRF1 and DYRK1A prognostic value.A substituição da válvula aórtica continua a ser a opção terapêutica de referência para doentes com estenose aórtica e visa a eliminação da sobrecarga de pressão, desencadeando a remodelagem reversa miocárdica. Contudo, apesar do benefício hemodinâmico imediato, nem todos os pacientes apresentam regressão completa da hipertrofia do miocárdio, ficando com maior risco de eventos adversos, como a insuficiência cardíaca. Atualmente, os mecanismos biológicos subjacentes a uma remodelagem reversa incompleta ainda não são claros. Além disso, não dispomos de ferramentas de prognóstico definitivos nem de terapias auxiliares para melhorar a condição dos pacientes indicados para substituição da válvula. Para ajudar a resolver estas lacunas, uma abordagem combinada de (fosfo)proteómica e proteómica para a caracterização, respetivamente, do miocárdio e do líquido pericárdico foi seguida, tomando partido de biópsias e líquidos pericárdicos recolhidos em ambiente cirúrgico. Das mais de 1800 e 750 proteínas identificadas, respetivamente, no miocárdio e no líquido pericárdico dos pacientes com estenose aórtica, um total de 90 proteínas desreguladas foram detetadas. As análises de anotação de genes, de enriquecimento de vias celulares e discriminativa corroboram um cenário de aumento da expressão de genes pro-hipertróficos e de síntese proteica, um sistema ubiquitina-proteassoma ineficiente, uma tendência para morte celular (potencialmente acelerada pela atividade do complemento e por outros fatores extrínsecos que ativam death receptors), com ativação da resposta de fase aguda e do sistema imune, assim como da fibrose. A validação de alguns alvos específicos através de immunoblot e correlação com dados clínicos apontou para a cadeia β do complemento C3, a Muscle Ring Finger protein 1 (MuRF1) e a dual-specificity Tyr-phosphoylation regulated kinase 1A (DYRK1A) como potenciais marcadores de uma resposta incompleta. Por outro lado, a predição de cinases a partir do fosfoproteoma, sugere que a modulação da caseína cinase 2, a família de cinases do IκB, a glicogénio sintase cinase 3 e da DYRK1A pode ajudar a melhorar a condição dos pacientes indicados para intervenção. Em particular, a avaliação funcional de cardiomiócitos DYRK1A+/- mostraram que esta cinase pode ser um alvo importante para tratar a disfunção cardíaca, uma vez que os miócitos mutantes responderam de forma diferente ao estiramento e mostraram uma menor capacidade para desenvolver força (tensão ativa). Este estudo levanta várias hipóteses na investigação da remodelagem reversa. No futuro, estudos de ganho e/ou perda de função realizados em cardiomiócitos isolados ou em modelos animais de banding-debanding da aorta ajudarão a testar a eficácia de modular os potenciais alvos terapêuticos encontrados. Além disso, estudos clínicos em coortes de maior dimensão trarão conclusões definitivas quanto ao valor de prognóstico do complemento C3, MuRF1 e DYRK1A.Programa Doutoral em Biomedicin

    Interdisciplinary Pain Rehabilitation Programs: Evidence and Clinical Real-World Results

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    Chronic pain conditions are influenced by and interact with physical, psychological, social, and contextual factors. These conditions are associated with psychological distress, poor health, sick leave, and high socio-economic costs. Therefore, modern clinical practice applies a biopsychosocial (BPS) framework. Interdisciplinary pain rehabilitation programs (IPRPs) for chronic pain distinguish themselves as well-coordinated complex interventions. This chapter describes the contents of such programs. We will briefly review the evidence for IPRPs and discuss problems when evaluating these complex interventions. Furthermore, we will report practice-based results from a large Swedish pain registry—the Swedish Quality Registry for Pain Rehabilitation (SQRP). The SQRP collects data from a relevant special clinical department in Sweden—i.e., real-life outcomes will be depicted. Characteristics of patients that benefit the most from IPRPs will be described and discussed. The indications for IPRPs will also be presented. Finally, we will discuss how to improve rehabilitation for chronic pain patients

    Desarrollo de una herramienta integral de gestión de gases de efecto invernadero para la toma de decisión contra el cambio climático a nivel regional y local en la Comunitat Valenciana

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    Tesis por compendio[ES] Actualmente, los responsables de tomar decisiones contra el cambio climático carecen de herramientas para desarrollar inventarios de emisiones de gases de efecto invernadero (GEI) con suficiente rigor científico-técnico y precisión para priorizar e invertir los recursos disponibles de manera eficiente en las medidas necesarias para luchar contra el cambio climático. Por ello, en esta tesis se expone el desarrollo de un sistema de información territorial y sectorial (SITE) para monitorear las emisiones de GEI que sirva como herramienta de gobernanza climática local y regional. SITE combina las ventajas de los enfoques metodológicos descendente o top-down (de arriba hacia abajo) y ascendente o bottom-up (de abajo hacia arriba), para lograr un enfoque híbrido innovador para contabilizar y gestionar de manera eficiente las emisiones de GEI. Por tanto, en esta tesis se definen los diferentes desarrollos metodológicos, tanto generales como específicos de sectores clave del Panel Intergubernamental de Cambio Climático (IPPC) (edificación, transporte, sector forestal, etc.), un desarrollo informático para la parte de SITE que se ejecuta del lado del servidor, que de ahora en adelante denominaremos back-end del sistema, y siete implementaciones como casos de estudio representativos, a diferentes escalas y aplicados sobre diferentes sectores. Estas implementaciones a diferentes escalas y sectores demuestran el potencial del sistema como herramienta de apoyo en la toma de decisión contra el cambio climático a nivel regional y local. Las diferentes implementaciones en casos piloto representativos, tanto a nivel regional en la Comunitat Valenciana como a nivel local en municipios grandes (València) y medianos (Quart de Poblet y Llíria) muestran el potencial de adaptación territorial y sectorial que tiene la herramienta. Las metodologías desarrolladas para los sectores específicos de tráfico rodado, edificación o sector forestal, ofrecen cuantificaciones con una resolución espacial con gran capacidad de optimizar las políticas locales y regionales. Por tanto, la herramienta cuenta con un gran potencial de escalabilidad y gran capacidad de mejora continua mediante la inclusión de nuevos enfoques metodológicos, adaptación de las metodologías a la disponibilidad de datos, metodologías concretas para sectores clave y actualización a las mejores metodologías disponibles derivadas de actividades de investigación de la comunidad científica.[CA] Actualment, els responsables de prendre decisions contra el canvi climàtic no tenen eines per aconseguir inventaris d'emissions de gasos d'efecte hivernacle (GEH) amb prou cientificotècnic rigor, precisió i integritat per invertir els recursos disponibles de manera eficient en les mesures necessàries contra el canvi climàtic. Per això, en aquesta tesis se exposa el desenvolupa un sistema d'informació territorial i sectorial (SITE) per monitoritzar les emissions de GEH com a eina de governança climàtica local i regional. Aquest sistema combina els avantatges dels enfocaments metodològics descendent o top-down (de dalt a baix) i ascendent o bottom-up (de baix a dalt), per aconseguir un enfocament híbrid innovador per comptabilitzar i gestionar de manera eficient les emissions de GEH. Per tant, en aquesta tesi doctoral es descriuen els diferents desenvolupaments metodològics, tant generals com específics de sectors clau del Panel Intergovernamental contra el Canvi Climàtic (edificació, transport, forestal, etc.), un desenvolupament informàtic per al back-end del sistema i set implementacions com a casos d'estudi representatius, a diferents escales, amb els diferents enfocaments metodològics i aplicats sobre diferents sectors. Això queda descrit en sis capítols. Aquestes implementacions a diferents escales i sectors demostren el potencial del sistema com a eina de suport en la presa de decisió contra el canvi climàtic a nivell regional i local. Les diferents implementacions en casos pilot representatius, tant a nivell regional a la Comunitat Valenciana com a nivell local en municipis grans (València) i mitjans (Quart de Poblet i Llíria,) mostren el potencial d'adaptació territorial i sectorial que té l'eina. Les metodologies desenvolupades per als sectors específics de trànsit rodat, edificació i forestal, ofereixen quantificacions amb una resolució espacial amb gran capacitat d'optimitzar les polítiques locals i regionals. Per tant, l'eina compta amb un gran potencial d'escalabilitat i gran capacitat de millora contínua mitjançant la inclusió de nous enfocaments metodològics, adaptació de les metodologies a la disponibilitat de dades, metodologies concretes per a sectors clau, i actualització a les millors metodologies disponibles derivades de activitats de investigació de la comunitat científica.[EN] Currently, regional and local decision-makers lack of tools to achieve greenhouse gases (GHG) emissions inventories with enough rigor, accuracy and completeness in order to prioritize available resources efficiently against climate change. Thus, in this thesis the development of a territorial and sectoral information system (SITE) to monitor GHG emissions as a local and regional climate governance tool is exposed. This system combines the advantages of both, top-down and bottom-up approaches, to achieve an innovative hybrid approach to account and manage efficiently GHG emissions. Furthermore, this thesis defines the methodologies developed, a computer proposal for the back-end of the system and seven implementations as representative case studies at different scales (local and regional level), with the different methodological approaches and applied to different sectors. Thus, these implementations demonstrate the potential of the system as decision-making tool against climate change at the regional and local level as climate governance tool. The different implementations in representative pilot cases, both at the regional level in the Valencian Community and at the local level in large (Valencia) and medium-sized municipalities (Quart de Poblet and Llíria) demonstrate the potential for territorial and sectoral adaptation of the system developed. The methodologies developed for the specific sectors of road transport, building and forestry, offer quantifications with a spatial resolution with a great capacity to optimize local and regional policies. Therefore, the tool has a great potential for scalability and a great capacity for continuous improvement through the inclusion of new methodological approaches, adapting the methodologies to the availability of data, specific methodologies for key sectors, and updating to the best methodologies available in the scientific community.Lorenzo Sáez, E. (2022). Desarrollo de una herramienta integral de gestión de gases de efecto invernadero para la toma de decisión contra el cambio climático a nivel regional y local en la Comunitat Valenciana [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/181662TESISCompendi

    Quantifying the Indirect Effect of Wolves on Aspen in Northern Yellowstone National Park: Evidence for a Trophic Cascade?

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    Yellowstone National Park is renowned for its incredible wildlife, and perhaps the most famous of these species is the gray wolf, which was reintroduced to the Park in the mid-1990s. After reintroduction, it was highly publicized by scientists, journalists, and environmentalists that the wolf both decreased elk density and changed elk behavior in a way that reduced elk effects on plants, a process known as a “trophic cascade.” Aspen, which is eaten by elk in winter, is one species at the forefront of Yellowstone trophic cascade research because it has been in decline across the Park for over a century. However, due to the challenges of measuring trophic cascades, there is continued uncertainty regarding the effects of wolves on aspen in northern Yellowstone. Thus, the purpose of my dissertation was to provide a comprehensive test of a trophic cascade in this system. Specifically, I used 20 years of data on aspen, elk, and wolves in Yellowstone to: 1) clarify annual trends in browsing and height of young aspen (a proxy for regeneration) after wolf reintroduction, 2) assess the influence of wolves scaring elk on aspen (“trait-mediated indirect effects”), and 3) evaluate the effect of wolves killing elk on aspen (“density-mediated indirect effects”). My research suggests that wolves indirectly contributed to increased aspen over story recruitment following their reintroduction by helping to reduce the elk population size, but elk response to the risk of wolf predation did not reduce elk foraging in a way that measurably increased aspen recruitment. Additionally, hunter harvest of elk north of the park was twice as important as wolf predation in causing increased aspen recruitment. However, despite wolves and hunters limiting elk abundance, it is still uncommon for young aspen to grow past peak browsing height (120-cm), indicating that many stands remain vulnerable to elk herbivory nearly 30 years after wolf reintroduction. These results highlight that the strength and mechanism of predator effects on plant communities are context-specific. Thus, using predator reintroduction as a tool for ecosystem restoration without considering the many factors that shape trophic cascades may result in different management and conservation outcomes than intended

    Células endoteliais progenitoras e células endoteliais circulantes na insuficiência cardíaca: um estudo transversal

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    The objective of the present thesis was to compare the levels of circulating endothelial progenitor cells (EPCs), circulating endothelial cells (CECs), and hematopoietic stem cells (HSCs) between patients with heart failure with reduced ejection fraction (HFrEF) and a group of subjects with cardiovascular risk factors. We also compared the levels of circulating EPCs, CECs, and HSCs between subgroups regarding the presence of cardiovascular risk factors (e.g. diabetes mellitus) and the etiology of heart failure (HF). To achieved this, whole peripheral blood was drawn from patients previously diagnosed with HFrEF (n = 42) and age-matched subjects presenting similar cardiovascular risk factors but without established cardiovascular disease (n = 42). Then, a combination of markers was used in peripheral blood samples in order to assess the number of circulating EPCs, CECs, and HSCs via flow cytometry analysis. Patients with HFrEF had significantly decreased levels of circulating EPCs (5.28 x 10-3 ± 6.83 x 10-4 % vs 7.76 x 10-3 ± 4.91 x 10-4 %, P ≤ 0.001) and CECs (5.11 x 10-3 ± 7.87 x 10-4 % vs 6.51 x 10-3 ± 5.21 x 10-4 %, P = 0.005) compared to subjects with cardiovascular risk factors. However, levels of HSCs were not significantly different between the two groups (P = 0.590). Additionally, CECs (6.69 x 10-3 ± 6.38 x 10-3 % vs 3.61 x 10-3 ± 2.71 x 10-3 %, P = 0.057) tended to circulate in higher number in patients with ischemic HF compared to patients with non-ischemic HF. Patients with HFrEF and diagnosed as overweight/obese had significantly higher levels of circulating EPCs (6.10 x 10-3 ± 4.78 x 10-3 % vs 4.13 x 10-3 ± 3.55 x 10-3 %, P = 0.043) and CECs (6.27 x 10-3 ± 5.66 x 10-3 % vs 3.47 x 10-3 ± 3.54 x 10-3 %, P = 0.019) when compared to patients with HFrEF presenting a normal weight. Lastly, when comparing subjects from the age-matched group, subjects with dyslipidemia had significantly higher levels of CECs (7.74 x 10-3 ± 3.64 x 10-3 % vs 5.34 x 10-3 ± 2.59 x 10-3 %, P = 0.042) compared to subjects without dyslipidemia. In conclusion, the main result of this study is that the circulating levels of EPCs and CECs were significantly decreased in patients with HFrEF in comparison to subjects with cardiovascular risk factors. The current observations regarding cardiovascular risk factors suggest that EPCs, CECs, and HSCs play an important role in the detection and repair of vascular damage and endothelial dysfunction.O presente trabalho teve como principal objetivo comparar os níveis de células endoteliais progenitoras (CEPs), células endoteliais circulantes (CECs) e células estaminais hematopoiéticas (CEHs) em circulação entre doentes com insuficiência cardíaca com fração de ejeção reduzida (ICFEr) e um grupo de adultos com fatores de risco cardiovasculares. Adicionalmente, os níveis das CEPs, CECs e CEHs foram comparados entre subgrupos em função da presença de fatores de risco (ex. diabetes) e da etiologia da insuficiência cardíaca. Inicialmente foram recolhidas amostras de sangue periférico de doentes com ICFEr (n = 42) e indivíduos da mesma faixa etária com fatores de risco cardiovasculares, mas sem qualquer doença cardiovascular estabelecida (n = 42). Em seguida, foi utilizada uma combinação de anticorpos nas amostras de sangue periférico para quantificação do número de CEPs, CECs e CEHs por citometria de fluxo. Doentes com ICFEr apresentaram níveis de CEPs (5.28 x 10-3 ± 6.83 x 10-4 % vs 7.76 x 10-3 ± 4.91 x 10-4 %, P ≤ 0.001) e CECs (5.11 x 10- 3 ± 7.87 x 10-4 % vs 6.51 x 10-3 ± 5.21 x 10-4 %, P = 0.005) significativamente inferiores aos indivíduos com fatores de risco cardiovasculares. Contudo, não foram encontradas diferenças significativas nos níveis de CEHs entre os dois grupos (P = 0.590). Adicionalmente, observou-se que as CECs (6.69 x 10-3 ± 6.38 x 10-3 % vs 3.61 x 10-3 ± 2.71 x 10-3 %, P = 0.057) tendem a circular em maior número em doentes com ICFEr com etiologia isquémica comparativamente a doentes com ICFEr não isquémica. Doentes com ICFEr e com sobrepeso/obesidade apresentaram níveis de CEPs (6.10 x 10-3 ± 4.78 x 10-3 % vs 4.13 x 10-3 ± 3.55 x 10-3 %, P = 0.043) e CECs (6.27 x 10-3 ± 5.66 x 10- 3 % vs 3.47 x 10-3 ± 3.54 x 10-3 %, P = 0.019) significativamente superiores comparativamente a doentes com ICFEr e com peso normal. Por último, dentro do grupo de indivíduos com fatores de risco cardiovasculares, indivíduos com dislipidemia apresentaram níveis de CECs (7.74 x 10-3 ± 3.64 x 10-3 % vs 5.34 x 10-3 ± 2.59 x 10-3 %, P = 0.042) significativamente superiores em comparação a indivíduos sem dislipidemia. Em conclusão, os principais resultados deste estudo indicam que o número de CECs e CEPs em circulação encontra-se significativamente reduzido em doentes com ICFEr comparativamente a indivíduos com fatores de risco para doenças cardiovasculares. As observações atuais em relação aos fatores de risco para doenças cardiovasculares sugerem que CEPs, CECs e CEHs desempenham um papel fundamental na sinalização e reparação do dano vascular e disfunção endotelial.Mestrado em Biomedicina Molecula
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