27,710 research outputs found

    Identifizierung prädiktiver und prognostischer Biomarker in unterschiedlichen Tumorkompartimenten des ösophagealen Adenokarzinoms

    Get PDF
    Das √∂sophageale Adenokarzinom zeigt eine global steigende Inzidenz und hat mit einer 5-Jahres-√úberlebensrate von weniger als 25% eine schlechte Prognose. Personalisierte Therapieans√§tze sind selten und prognostische/pr√§diktive Biomarker des Tumormikromilieus sind unzureichend charakterisiert. Die kumulative Promotion n√§hert sich dieser Problematik in drei unterschiedlichen Schwerpunkten. 1. Zur Identifizierung Kompartiment-spezifischer Biomarker wurde eine Methode entwickelt, welche als kosteng√ľnstige Alternative zum sc-Seq Expressionsprofile individueller Zelltypen generiert. Dabei erfolgt die Extraktion der RNA nicht aus Einzelzellen, sondern aus flowzytometrisch-getrennten Zellkompartimenten. Die Separation der Proben in Epithelzellen, Immunzellen und Fibroblasten wurde durch verschiedene Verfahren validiert und eine suffiziente Ausbeute an RNA auch f√ľr kleine Gewebemengen gezeigt. 2. Biomarker des Immunzellkompartiments als therapeutische Angriffspunkte wurden in einem Patientenkollektiv von bis zu 551 Patienten auf ihre Bedeutung beim EAC √ľberpr√ľft. Es zeigte sich eine Expression der Immuncheckpoints LAG3, VISTA und IDO auf TILs durch IHC und RNA-Sonden basierte Verfahren in einem relevanten Anteil (LAG3: 11,4%, VISTA: 29%, IDO: 52,6%). Es konnte eine prognostisch g√ľnstige Bedeutung der VISTA, LAG3 und IDO Expression gezeigt werden. Durch den Vergleich von Genexpressionsprofilen aus therapienaiven und vorbehandelten Tumoren konnte zudem ein immunsuppressiver Effekt von neoadjuvanten Therapiekonzepten auf das Tumormikromilieu des EACs gezeigt werden. Dabei kam es zur verminderten Expression von Checkpoints und Anzahl TILs nach (Radio-) Chemotherapie. 3. Im Tumorzellkompartiment wurde die Rolle von Amplifikationen in ErbB-Rezeptor abh√§ngigen Signalwegen durch FISH-Technik und Immunhistochemie evaluiert. Es fanden sich KRAS Amplifikationen in 17,1%, PIK3CA Amplifikationen in 5% sowie eine HER2/neu-√úberexpression in 14,9% der untersuchten Tumore

    Die akute Appendizitis im Kindes- und Jugendalter: neue diagnostische Verfahren f√ľr die pr√§therapeutische Differenzierung histopathologischer Entit√§ten zur Unterst√ľtzung konservativer Therapiestrategien

    Get PDF
    Hintergrund der hier zusammengefassten Studien war die aktuelle Datenlage, die daf√ľr spricht, dass es sich bei der klinisch unkomplizierten, histopathologisch phlegmon√∂sen und der klinisch komplizierten, histopathologisch gangr√§n√∂sen Appendizitis um unabh√§ngige Entit√§ten handelt. Diese k√∂nnen unterschiedlichen Therapieoptionen (konservativ vs. operativ) zugef√ľhrt werden. Vor diesem Hintergrund war es ein Ziel der Arbeiten zu untersuchen, wie die Formen der akuten Appendizitis im Kindes- und Jugendalter bereits pr√§therapeutisch unterschieden werden k√∂nnen. Sowohl in der Labordiagnostik (P1 und P2) als auch im Ultraschall (P3) lassen sich Unterschiede zwischen Patient*innen mit unkomplizierter, phlegmon√∂ser und komplizierter (gangr√§n√∂ser und perforierender) Appendizitis aufzeigen. Hierdurch allein kann allerdings aufgrund unzureichender Trennsch√§rfe noch keine ausreichende Entscheidungssicherheit erreicht werden. Mit Verfahren der k√ľnstlichen Intelligenz auf Untersucher-unabh√§ngige diagnostische Parameter (P4) konnte die Vorhersagegenauigkeit der akuten Appendizitis weiter gesteigert werden. Interessante Ergebnisse bez√ľglich der unterschiedlichen Pathomechanismen der beiden inflammatorischen Entit√§ten ergaben sich durch eine differenzielle Genexpressionsanalyse (P5). In einer Proof-of-Concept-Studie wurden zuvor beschriebene Methoden der k√ľnstlichen Intelligenz auf die Genexpressionsdaten angewandt (P6). Hierdurch konnte im Modell eine grunds√§tzliche Differenzierbarkeit der Entit√§ten durch die Anwendung der neuen Methode aufgezeigt werden. Ein mittelfristiges Ziel ist es, eine Biomarkersignatur zu definieren, die ihre Aussagekraft durch einen Computeralgorithmus hat. Hierdurch soll eine schnelle Therapieentscheidung erm√∂glicht werden. Im Idealfall sollte diese Biomarkersignatur sicher, objektiv und einfach zu bestimmen sein sowie eine h√∂here diagnostische Sicherheit als die bisherige Diagnostik mittels Anamnese, Untersuchung, Laboranalyse und Ultraschall bieten. Langfristiges Ziel von Folgestudien ist die Identifizierung einer Biomarkersignatur mit der bestm√∂glichen Vorhersagekraft. Hinsichtlich der routinem√§√üigen klinischen Diagnostik ist die Anwendung von Point-of-Care Devices auf PCR-Basis denkbar. Hier k√∂nnte eine limitierte Anzahl von Primern f√ľr eine Biomarkersignatur mit hoher Vorhersagekraft zum Einsatz kommen. Der dadurch ermittelte Biomarker w√ľrde seine Aussagekraft durch einen einfach anzuwendenden Computeralgorithmus erhalten. Die Kombination aus Genexpressionsanalyse mit Methoden der k√ľnstlichen Intelligenz kann somit die Grundlage f√ľr ein neues diagnostisches Instrument zur sicheren Unterscheidung unterschiedlicher Appendizitisentit√§ten darstellen

    Immunosuppressive drugs in renal transplantation

    Get PDF
    A kidney transplant, sometimes known as a renal transplant, is the treatment of choice for kidney failure at end stage renal disease (ESRD). The renal transplant surgery is followed by a lifetime course of immunosuppressive agents, divided into initial induction phase and later maintenance phase. It is seen that the risk of acute rejection is maximum in the initial months after transplantation (induction phase) and then reduces later (maintenance phase). In induction phase there is use of high-intensity immunosuppression immediately after transplantation, when the risk of rejection is maximum and then the dose reduced for long- term therapy. The main challenge in the renal transplantation community is long- term transplant survival. Long-term graft loss is mainly due to acute and chronic graft rejection, and also due to complications of immunosuppressive therapy. Currently, there is triple therapy as conventional immunosuppressive protocol: a calcineurin inhibitor, an antimetabolite agent, and a corticosteroid. The main aim of development of new immunosuppressive agents is not only improvement of short- term outcomes but also to increase the long- term graft survival by less nephrotoxicity, and minimal side-effects

    Anu√°rio cient√≠fico da Escola Superior de Tecnologia da Sa√ļde de Lisboa - 2021

    Get PDF
    √Č com grande prazer que apresentamos a mais recente edi√ß√£o (a 11.¬™) do Anu√°rio Cient√≠fico da Escola Superior de Tecnologia da Sa√ļde de Lisboa. Como institui√ß√£o de ensino superior, temos o compromisso de promover e incentivar a pesquisa cient√≠fica em todas as √°reas do conhecimento que contemplam a nossa miss√£o. Esta publica√ß√£o tem como objetivo divulgar toda a produ√ß√£o cient√≠fica desenvolvida pelos Professores, Investigadores, Estudantes e Pessoal n√£o Docente da ESTeSL durante 2021. Este Anu√°rio √©, assim, o reflexo do trabalho √°rduo e dedicado da nossa comunidade, que se empenhou na produ√ß√£o de conte√ļdo cient√≠fico de elevada qualidade e partilhada com a Sociedade na forma de livros, cap√≠tulos de livros, artigos publicados em revistas nacionais e internacionais, resumos de comunica√ß√Ķes orais e p√≥steres, bem como resultado dos trabalhos de 1¬ļ e 2¬ļ ciclo. Com isto, o conte√ļdo desta publica√ß√£o abrange uma ampla variedade de t√≥picos, desde temas mais fundamentais at√© estudos de aplica√ß√£o pr√°tica em contextos espec√≠ficos de Sa√ļde, refletindo desta forma a pluralidade e diversidade de √°reas que definem, e tornam √ļnica, a ESTeSL. Acreditamos que a investiga√ß√£o e pesquisa cient√≠fica √© um eixo fundamental para o desenvolvimento da sociedade e √© por isso que incentivamos os nossos estudantes a envolverem-se em atividades de pesquisa e pr√°tica baseada na evid√™ncia desde o in√≠cio dos seus estudos na ESTeSL. Esta publica√ß√£o √© um exemplo do sucesso desses esfor√ßos, sendo a maior de sempre, o que faz com que estejamos muito orgulhosos em partilhar os resultados e descobertas dos nossos investigadores com a comunidade cient√≠fica e o p√ļblico em geral. Esperamos que este Anu√°rio inspire e motive outros estudantes, profissionais de sa√ļde, professores e outros colaboradores a continuarem a explorar novas ideias e contribuir para o avan√ßo da ci√™ncia e da tecnologia no corpo de conhecimento pr√≥prio das √°reas que comp√Ķe a ESTeSL. Agradecemos a todos os envolvidos na produ√ß√£o deste anu√°rio e desejamos uma leitura inspiradora e agrad√°vel.info:eu-repo/semantics/publishedVersio

    Neuroanatomical and gene expression features of the rabbit accessory olfactory system. Implications of pheromone communication in reproductive behaviour and animal physiology

    Get PDF
    Mainly driven by the vomeronasal system (VNS), pheromone communication is involved in many species-specific fundamental innate socio-sexual behaviors such as mating and fighting, which are essential for animal reproduction and survival. Rabbits are a unique model for studying chemocommunication due to the discovery of the rabbit mammary pheromone, but paradoxically there has been a lack of knowledge regarding its VNS pathway. In this work, we aim at filling this gap by approaching the system from an integrative point of view, providing extensive anatomical and genomic data of the rabbit VNS, as well as pheromone-mediated reproductive and behavioural studies. Our results build strong foundation for further translational studies which aim at implementing the use of pheromones to improve animal production and welfare

    Antioxidation, Anti-Inflammation, and Regulation of SRD5A Gene Expression of Oryza sativa cv. Bue Bang 3 CMU Husk and Bran Extracts as Androgenetic Alopecia Molecular Treatment Substances

    Get PDF
    Acknowledgments: The authors are grateful to the NRCT for supporting research facilities (grant no. NRCT5-RRI63004-P05), Chiang Mai University for the Fundamental Fund 2022, and the partially support grant. We would like to thank Lanna Rice Research Center, Chiang Mai University, and Saleekam Trading Co., Ltd., Thailand, for providing the rice bran and husk samples.Data Availability Statement: The data presented in this study are available on request from the corresponding author.Funding: This research project is supported by National Research Council of Thailand (NRCT): NRCT5-RRI63004-P05, Fundamental Fund 2022, Chiang Mai University, and partially supported by Chiang Mai University.Androgenetic alopecia (AGA), a hair loss disorder, is a genetic predisposition to sensitive androgens, inflammation, and oxidative stress. Unfortunately, current treatments with synthetic medicines contain a restricted mechanism along with side effects, whereas the bioactive constituents of plant extracts are multifunctional, with fewer side effects. The massive amounts of rice husk and bran are agricultural wastes that may cause pollution and environmental problems. Owing to these rationales, the local rice variety, Bue Bang 3 CMU (BB3CMU), which is grown in northern Thailand, was evaluated for the valuable utilization of rice by-products, husk (BB3CMU-H) and bran (BB3CMU-RB) extracts, for AGA treatment regarding antioxidant, anti-inflammatory, anti-androgenic activities, and the characterization of bioactive compounds. Our study verified that BB3CMU-H had the highest level of polyphenols, contributing to its greater antioxidant activity. Conversely, BB3CMU-RB was the predominant source of tocopherols, resulting in better anti-androgenic activities regarding the downregulation of steroid 5őĪ-reductase genes (SRD5A). Notably, anti-inflammation via the attenuation of nitric oxide productions was observed in BB3CMU-H (0.06 ¬Ī 0.13 őľM) and BB3CMU-RB (0.13 ¬Ī 0.01 őľM), which were significantly comparable to diclofenac sodium salt (0.13 ¬Ī 0.19 őľM). Therefore, the combination of BB3CMU-H and BB3CMU-RB could be utilized in cosmeceutical and pharmaceutical applications for AGA patientsNational Research Council of Thailand (NRCT): NRCT5-RRI63004-P05Fundamental Fund 2022Chiang Mai Universit

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

    Get PDF
    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 ő≤ 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 IőļB 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 substitui√ß√£o da v√°lvula a√≥rtica continua a ser a op√ß√£o terap√™utica de refer√™ncia para doentes com estenose a√≥rtica e visa a elimina√ß√£o da sobrecarga de press√£o, desencadeando a remodelagem reversa mioc√°rdica. Contudo, apesar do benef√≠cio hemodin√Ęmico imediato, nem todos os pacientes apresentam regress√£o completa da hipertrofia do mioc√°rdio, ficando com maior risco de eventos adversos, como a insufici√™ncia card√≠aca. Atualmente, os mecanismos biol√≥gicos subjacentes a uma remodelagem reversa incompleta ainda n√£o s√£o claros. Al√©m disso, n√£o dispomos de ferramentas de progn√≥stico definitivos nem de terapias auxiliares para melhorar a condi√ß√£o dos pacientes indicados para substitui√ß√£o da v√°lvula. Para ajudar a resolver estas lacunas, uma abordagem combinada de (fosfo)prote√≥mica e prote√≥mica para a caracteriza√ß√£o, respetivamente, do mioc√°rdio e do l√≠quido peric√°rdico foi seguida, tomando partido de bi√≥psias e l√≠quidos peric√°rdicos recolhidos em ambiente cir√ļrgico. Das mais de 1800 e 750 prote√≠nas identificadas, respetivamente, no mioc√°rdio e no l√≠quido peric√°rdico dos pacientes com estenose a√≥rtica, um total de 90 prote√≠nas desreguladas foram detetadas. As an√°lises de anota√ß√£o de genes, de enriquecimento de vias celulares e discriminativa corroboram um cen√°rio de aumento da express√£o de genes pro-hipertr√≥ficos e de s√≠ntese proteica, um sistema ubiquitina-proteassoma ineficiente, uma tend√™ncia para morte celular (potencialmente acelerada pela atividade do complemento e por outros fatores extr√≠nsecos que ativam death receptors), com ativa√ß√£o da resposta de fase aguda e do sistema imune, assim como da fibrose. A valida√ß√£o de alguns alvos espec√≠ficos atrav√©s de immunoblot e correla√ß√£o com dados cl√≠nicos apontou para a cadeia ő≤ 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 predi√ß√£o de cinases a partir do fosfoproteoma, sugere que a modula√ß√£o da case√≠na cinase 2, a fam√≠lia de cinases do IőļB, a glicog√©nio sintase cinase 3 e da DYRK1A pode ajudar a melhorar a condi√ß√£o dos pacientes indicados para interven√ß√£o. Em particular, a avalia√ß√£o funcional de cardiomi√≥citos DYRK1A+/- mostraram que esta cinase pode ser um alvo importante para tratar a disfun√ß√£o card√≠aca, uma vez que os mi√≥citos mutantes responderam de forma diferente ao estiramento e mostraram uma menor capacidade para desenvolver for√ßa (tens√£o ativa). Este estudo levanta v√°rias hip√≥teses na investiga√ß√£o da remodelagem reversa. No futuro, estudos de ganho e/ou perda de fun√ß√£o realizados em cardiomi√≥citos isolados ou em modelos animais de banding-debanding da aorta ajudar√£o a testar a efic√°cia de modular os potenciais alvos terap√™uticos encontrados. Al√©m disso, estudos cl√≠nicos em coortes de maior dimens√£o trar√£o conclus√Ķes definitivas quanto ao valor de progn√≥stico do complemento C3, MuRF1 e DYRK1A.Programa Doutoral em Biomedicin

    Estudo de altera√ß√Ķes gen√©ticas no NAPRT e NAMPT em cancro

    Get PDF
    NAD+ is both a co-enzyme for oxidation-reduction reactions and a substrate for NAD+ -consuming enzymes and therefore, it is critical for many cellular processes. In mammalian cells, intracellular NAD+ can be synthesized through either de novo synthesis, from tryptophan, or via salvage pathways, from other precursors, such as NAM and NA which are converted by NAMPT and NAPRT, respectively. Changes in the NAD+ content in cells and tissues are linked with a wide variety of diseases, including cancer. A large proportion of cancer cases is still diagnosed only at an advanced stage and thus, there is a need for new affordable, specific, sensitive and non-invasive biomarkers. Salivary biomarkers can meet these criteria and thus, are promising tools in cancer screening, diagnostic and prognostic. The main objective of this work was to study genetic alterations in NAMPT and NAPRT in DNA samples from healthy donors and in samples from cancer patients. For this, DNA and RNA extraction from saliva samples was optimized, as a starting point to study this biofluid as a source for cancer biomarkers. The results from the bioinformatics analysis showed that the frequency of alterations in NAPRT and NAMPT genes is low in the cancer contexts investigated. Nevertheless, it is still necessary to further study the impact that these alterations might have. There is also a great need to investigate and optimize methods for saliva studies, in order to promote it as a liquid biopsy of regular use in clinical settings.O NAD+ funciona como coenzima em rea√ß√Ķes de oxida√ß√£o-redu√ß√£o e como um substrato para determinadas enzimas e tem, portanto, um papel cr√≠tico em muitos processos celulares. Nas c√©lulas de mam√≠feros, o NAD+ intracelular pode ser sintetizado atrav√©s da s√≠ntese de novo, a partir do triptofano, ou atrav√©s das vias de ‚Äúreciclagem‚ÄĚ, a partir de outros precursores, como NAM e NA, que s√£o convertidos por NAMPT e NAPRT, respetivamente. Altera√ß√Ķes no conte√ļdo de NAD+ em c√©lulas e tecidos est√£o relacionadas com uma ampla variedade de doen√ßas, incluindo cancro. Uma grande propor√ß√£o dos casos de tumores ainda √© diagnosticada j√° num estadio avan√ßado e, por isso, s√£o necess√°rios novos biomarcadores economicamente acess√≠veis, espec√≠ficos, sens√≠veis e n√£o invasivos. Os biomarcadores salivares conseguem cumprir esses crit√©rios e s√£o, assim, mol√©culas promissoras para o rastreio, diagn√≥stico e progn√≥stico de cancro. O principal objetivo deste trabalho foi estudar altera√ß√Ķes gen√©ticas no NAMPT e NAPRT em amostras de DNA de dadores saud√°veis e em amostras de pacientes com cancro. Para isto, foi otimizada a extra√ß√£o de DNA e RNA a partir de amostras de saliva, como ponto de partida para estudar este biofluido como fonte de biomarcadores de cancro. Os resultados da an√°lise bioinform√°tica mostraram que a frequ√™ncia de altera√ß√Ķes nos genes NAPRT e NAMPT √© baixa nos contextos de cancro investigados. Ainda assim, ser√£o necess√°rios mais estudos para analisar o impacto que estas altera√ß√Ķes poder√£o ter. H√° tamb√©m uma grande necessidade de investigar e otimizar m√©todos para estudos em saliva, a fim de promov√™-la como bi√≥psia l√≠quida de uso generalizado em ambiente cl√≠nico.Mestrado em Biomedicina Molecula
    • ‚Ķ
    corecore