949 resultados para Gut
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Dipeptidyl peptidase 4 (DPP-4) enzymatically inactivates incretin hormones, and DPP-4 inhibitor drugs are clinically approved therapies for type 2 diabetes. The primary substrates of DPP-4 are produced in the intestinal lining and we therefore investigated whether lactobacilli colonizing the gut can inhibit this enzyme. Fifteen Lactobacillus strains (Lb 1-15) from human infant faecal samples were isolated, identified, extracted and screened for inhibitory activity against DPP-4. Activity was compared against Lactobacillus reference strains (Ref 1-7), a Gram positive control (Ctrl 1) and two Gram negative controls (Ctrl 2-3). A range of DPP-4 inhibitory activity was observed (10-32%; P<0.05-0.001). Strains of L. fabifermentans (25%), L. plantarum (12-24%) and L. fermentum (14%) had significant inhibitory activity. However, we also noted that E. coli (Ctrl 2) and S. Typhimurium (Ctrl 3) had the greatest inhibitory activity (30-32%). Contrastingly, some isolates (Lb 12-15) and reference cultures (Ref 1-4) instead of inhibiting DPP-4 actually enhanced it, perhaps indicating the presence of X-prolyl-dipeptidyl-amino-peptidase (PepX). This provides a future rationale for using probiotic bacteria or their components for management of type 2 diabetes via DPP-4 inhibition.
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RATIONALE: Anaerobic bacteria are present in large numbers in the airways of people with cystic fibrosis (PWCF). In the gut, anaerobes produce short-chain fatty acids (SCFAs) that modulate immune/inflammatory processes.
OBJECTIVES: To investigate the capacity of anaerobes to contribute to CF airway pathogenesis via SCFAs.
METHODS: Samples from 109 PWCF were processed using anaerobic microbiological culture with bacteria present identified by 16S RNA sequencing. SCFAs levels in anaerobe supernatants and bronchoalveolar lavage (BAL) were determined by gas chromatography. The mRNA and/or protein expression of SCFAs receptors, GPR41 and GPR43, in CF and non-CF bronchial brushings, and 16HBE14o- and CFBE41o- cells were evaluated using RT-PCR, western blot, laser scanning cytometry and confocal microscopy. SCFAs-induced IL-8 secretion was monitored by ELISA.
MEASUREMENTS AND MAIN RESULTS: Fifty seven of 109 (52.3%) PWCF were anaerobe-positive. Prevalence increased with age, from 33.3% to 57.7% in PWCF under (n=24) and over 6 years (n=85). All evaluated anaerobes produced millimolar concentrations of SCFAs, including acetic, propionic and butyric acid. SCFAs levels were higher in BAL samples from adults than children. GPR41 levels were elevated in; CFBE41o- versus 16HBE14o- cells; CF versus non-CF bronchial brushings; 16HBE14o- cells after treatment with CFTR inhibitor CFTR(inh)-172, CF BAL, or inducers of endoplasmic reticulum stress. SCFAs induced a dose-dependent and pertussis toxin-sensitive IL-8 response in bronchial epithelial cells with a higher production of IL-8 in CFBE41o- than 16HBE14o- cells.
CONCLUSIONS: This study illustrates that SCFAs contribute to excessive production of IL-8 in CF airways colonized with anaerobes via upregulated GPR41.
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A 12 amino acid sequence from the adenovirus 12 E1B protein is homologous at the protein level with a similar 12-mer derived from the wheat protein A-gliadin. It has been suggested that exposure to Ad 12 could sensitise individuals to gliadins with resultant gluten sensitive enteropathy. In this study, the polymerase chain reaction (PCR) was used to analyse duodenal biopsy tissue from patients with coeliac disease for the presence of Ad 12. The sensitivity of the assay system was at least 1 in 10(5) cells and specificity was confirmed both by probing with an internal oligonucleotide and by direct sequencing. Ad 12 sequences were detected in three of 17 patients with adult coeliac disease and in five of 16 adult controls with normal duodenal biopsies. Since exposure to the virus would be predicted to occur in infancy we also studied patients with childhood coeliac disease diagnosed at less than 1 year of age. Ad 12 was positive in three of 10 childhood coeliac patients and one of seven controls. In addition, we studied a cohort of patients who presented with a diarrhoeal illness and associated anti alpha gliadin antibodies in 1983. These patients had duodenal biopsies performed at this time. One of three patients with abnormal histology had detectable Ad 12 while two of 14 with normal findings were positive for Ad 12. Finally, the potential oncogenic nature of Ad 12 prompted examination of a group of patients with intestinal tumours. Ad 12 DNA was, however, in only two of 19 tumour samples tested. These data indicate that Ad 12 can be successfully detected using PCR on paraffin embedded tissue. Furthermore, Ad 12 was detected at a relatively high level in normal duodenum. The results do not, however, support the hypothesis that prior exposure to Ad 12 is implicated in the pathogenesis of coeliac disease.
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A 3-year old child with juvenile chronic myeloid leukaemia received a T cell-depleted BMT from a male unrelated donor. There was early graft failure associated with increasing splenomegaly and hypersplenism. Splenectomy was performed 53 days post-transplant and was followed by autologous marrow recovery with return of leukaemia. A second unrelated donor BMT was performed 9 months later using T cell-replete marrow from a similarly matched female donor. Grade 2 GVHD involving the skin and gut responded to treatment with steroids. Chimaerism was assessed using Y-specific polymerase chain reaction (PCR) and microsatellites. Samples taken at the time of splenectomy showed no donor marrow engraftment but there was significant engraftment in the spleen. Following the second transplant, donor-type haematopoiesis was documented using a panel of microsatellite probes. The patient remains well 6 months after transplant. Splenectomy should be considered prior to transplant in patients with significant splenomegaly and hypersplenism. Partial chimaerism in the spleen, but not bone marrow, post-BMT, has not previously been documented. PCR technology is a useful and highly sensitive way to assess chimaerism post-BMT and is informative in sex-matched cases, whilst the small amount of material required is advantageous in paediatric patients.
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Understanding the dietary consumption and selection of wild populations of generalist herbivores is hampered by the complex array of factors. Here, we determine the influence of habitat, season, and animal density, sex, and age on the diet consumption and selection of 426 red deer (Cervus elaphus scoticus) culled in Fiordland National Park, New Zealand. Our site differs from studies elsewhere both in habitat (evergreen angiosperm-dominated forests) and the intensity of hunting pressures. We predicted that deer would not consume forage in proportion to its relative availability, and that dietary consumption would change among and within years in response to hunting pressures that would also limit opportunities for age and sex segregation. Using canonical correspondence analysis, we evaluated the relative importance of different drivers of variation in diet consumption assessed from gut content and related these to available forage in the environment. We found that altitude explained the largest proportion of variation in diet consumption, reflecting the ability of deer to alter their consumption and selection in relation to their foraging grounds. Grasses formed a high proportion of the diet consumption, even for deer culled several kilometres from the alpine grasslands. In the winter months, when the alpine grasslands were largely inaccessible, less grass was eaten and deer resorted to woody plants that were avoided in the summer months. Surprisingly, there were no significant dietary differences between adults and juveniles and only subtle differences between the sexes. Sex-based differences in diet consumption are commonly observed in ungulate species and we suggest that they may have been reduced in our study area owing to decreased heterogeneity in available forage as the diversity of palatable species decreased under high deer browsing pressures, or by intense hunting pressure. © 2009 The Authors. Journal compilation © 2009 Ecological Society of Australia.
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Rationale: IL-17A is purported to help drive early pathogenesis in acute respiratory distress syndrome (ARDS) by enhancing neutrophil recruitment. Whilst IL-17A is the archetypal cytokine of T helper (Th)17 cells, it is produced by a number of lymphocytes, the source during ARDS being unknown.
Objectives: To identify the cellular source and the role of IL17A in the early phase of lung injury
Methods: Lung injury was induced in WT (C57BL/6) and IL-17 KO mice with aerosolised LPS (100 µg) or Pseudomonas aeruginosa infection. Detailed phenotyping of the cells expressing RORγt, the transcriptional regulator of IL-17 production, in the mouse lung at 24 hours was carried out by flow cytometry.
Measurement and Main Results: A 100-fold reduction in neutrophil infiltration was observed in the lungs of the IL-17A KO compared to wild type (WT) mice. The majority of RORγt+ cells in the mouse lung were the recently identified type 3 innate lymphoid cells (ILC3). Detailed characterisation revealed these pulmonary ILC3s (pILC3s) to be discrete from those described in the gut. The critical role of these cells was verified by inducing injury in Rag2 KO mice which lack T cells but retain ILCs. No amelioration of pathology was observed in the Rag2 KO mice.
Conclusions: IL-17 is rapidly produced during lung injury and significantly contributes to early immunopathogenesis. This is orchestrated largely by a distinct population of pILC3 cells. Modulation of pILC3s’ activity may potentiate early control of the inflammatory dysregulation seen in ARDS, opening up new therapeutic targets.
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This article documents the public availability of (i) microbiomes in diet and gut of larvae from the dipteran Dilophus febrilis using massive parallel sequencing, (ii) SNP and SSR discovery and characterization in the transcriptome of the Atlantic mackerel (Scomber scombrus, L) and (iii) assembled transcriptome for an endangered, endemic Iberian cyprinid fish (Squalius pyrenaicus).
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OBJECTIVE: Studies indicate an inverse association between ductal adenocarcinoma of the pancreas (PDAC) and nasal allergies. However, controversial findings are reported for the association with asthma. Understanding PDAC risk factors will help us to implement appropriate strategies to prevent, treat and diagnose this cancer. This study assessed and characterised the association between PDAC and asthma and corroborated existing reports regarding the association between allergies and PDAC risk.
DESIGN: Information about asthma and allergies was collated from 1297 PDAC cases and 1024 controls included in the PanGenEU case-control study. Associations between PDAC and atopic diseases were studied using multilevel logistic regression analysis. Meta-analyses of association studies on these diseases and PDAC risk were performed applying random-effects model.
RESULTS: Asthma was associated with lower risk of PDAC (OR 0.64, 95% CI 0.47 to 0.88), particularly long-standing asthma (>=17 years, OR 0.39, 95% CI 0.24 to 0.65). Meta-analysis of 10 case-control studies sustained our results (metaOR 0.73, 95% CI 0.59 to 0.89). Nasal allergies and related symptoms were associated with lower risk of PDAC (OR 0.66, 95% CI 0.52 to 0.83 and OR 0.59, 95% CI 0.46 to 0.77, respectively). These results were supported by a meta-analysis of nasal allergy studies (metaOR 0.6, 95% CI 0.5 to 0.72). Skin allergies were not associated with PDAC risk.
CONCLUSIONS: This study shows a consistent inverse association between PDAC and asthma and nasal allergies, supporting the notion that atopic diseases are associated with reduced cancer risk. These results point to the involvement of immune and/or inflammatory factors that may either foster or restrain pancreas carcinogenesis warranting further research to understand the molecular mechanisms driving this association.
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Nas últimas décadas tem-se assistido a uma preocupação crescente relativamente às possíveis consequências da exposição a compostosxenóbioticos capazes de modular ou causar disrupção do sistema endócrino, os denominados Compostos Disruptores Endócrinos (CDEs). A maioria dos estudos efectuados tem-se centrado principalmente nos efeitos dos CDEs em vertebrados, enquanto que os seus efeitos em invertebrados têmsido negligenciados, embora este grupo represente mais de 95% de todas asespécies animais. Isópodes como o Porcellio scaber, combinam características associadas às mudas e aos processos reprodutivos mediados por mecanismos endócrinosconhecidos com um modo de vida terrestre, tornando-os potenciais espécies sentinela para estudos de disrupção endócrina (DE) em ambientes terrestres. Neste estudo, isópodes machos adultos, machos e fêmeas juvenis e casais foram expostos a concentrações crescentes dedois CDEs, vinclozolina (Vz) e bisfenol A (BPA). Testou-se a hipótese nula que a Vz e o BPA não interferem com o desenvolvimento e reprodução deste isópode terrestre. Foi investigadaa possível ligação entre os efeitos causados pelos compostos propostos e DE assim como a ligação a outros potenciais mecanismos de toxicidade.Parâmetros como concentração de 20-hidroxiecdisona (20E), muda, crescimento, rácios sexuais ediversos parâmetros reprodutivos foram estudados. Adicionalmente, de modo a estudar os alvos moleculares destes tóxicos, analisou-se a expressão proteica do intestino, hepatopâncreas e testículos do isópode após exposição aos químicos. Os resultados demonstram que a Vz e o BPA estimulam o aumento dos níveis de 20E de um modo dependente da dose. Excepção feita para a concentração mais baixa de BPA testada (10 mg/kg solo), para a qual concentrações significativamente mais altas de 20E foram determinadas, sugerindo a ocorrência dos “efeitos de baixas doses típicos de DE” já demonstrados por outros autores. O BPA também distorceu o rácio sexual favorecendo asfêmeas na concentração mais baixa. A mortalidade devido à ecdise incompleta foi relacionada com o hiper-ecdisonismo nas concentrações mais elevadas de Vz. Mais ainda, a Vz tende a atrasar a muda e o BPA a induzi-la. Não obstante, ambos os compostos provocam toxicidade no desenvolvimento,uma vez que foi encontrada uma diminuição generalizada nos parâmetros de crescimento. Os juvenis mostraram ser mais sensíveis à exposição aos tóxicos que os adultos. Estes compostos provocaram ainda toxicidade reprodutiva, com um decréscimo generalizado do “output” reprodutivo. A toxicidadecausada pelos ecdisteróides e o seu papel na síntese de vitelogenina são alguns dos factores chave que poderão influenciar negativamente a reprodução.A Vz e o BPA afectaram a expressão de proteínas envolvidas nometabolismo energético e induziram várias respostas de stress. Interferiram ainda com proteínas intimamente ligadas com o sucesso reprodutivo. Conclui-se assim,que ambos os CDEs propostos provocam toxicidade nodesenvolvimento e na reprodução de P. scaber, tendo sido evidenciada umaligação a DE. Alvos moleculares de natureza não-endócrina foram também revelados, através da expressão diferencial de algumas proteínaspreviamente descritas para invertebrados aquáticos e mesmo alguns vertebrados.
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The mechanisms of secretory granule biogenesis and regulated secretion of digestive enzymes in pancreatic acinar cells are still not well understood. To shed light on these processes, which are of biological and clinical importance (e.g., pancreatitis), a better molecular understanding of the components of the granule membrane, their functions and interactions is required. The application of proteomics has largely contributed to the identification of novel zymogen granule (ZG) proteins but was not yet accompanied by a better characterization of their functions. In this study we aimed at a) isolation and identification of novel membrane-associated ZG proteins; b) characterization of the biochemical properties and function of the secretory lectin ZG16p, a membrane-associated protein; c) exploring the potential of ZG16p as a new tool to label the endolysosomal compartment. First, we have performed a suborganellar proteomics approach by combining protein analysis by 2D-PAGE and identification by mass spectrometry, which has led to the identification of novel peripheral ZGM proteins with proteoglycan-binding properties (e.g., chymase, PpiB). Then, we have unveiled new molecular properties and (multiple) functions of the secretory lectin ZG16p. ZG16p is a unique mammalian lectin with glycan and proteoglycan binding properties. Here, I revealed for the first time that ZG16p is highly protease resistant by developing an enterokinase-digestion assay. In addition I revealed that ZG16p binds to a high molecular weight complex at the ZGM (which is also protease resistant) and forms highly stable dimers. In light of these findings I suggest that ZG16p is a key component of a predicted submembranous granule matrix attached to the luminal side of the ZGM that fulfils important functions during sorting and packaging of zymogens. ZG16p, may act as a linker between the matrix and aggregated zymogens due to dimer formation. Furthermore, ZG16p protease resistance might be of higher importance after secretion since it is known that ZG16p binds to pathogenic fungi in the gut. I have further investigated the role of ZG16p binding motifs in its targeting to ZG in AR42J cells, a pancreatic model system. Point mutations of the glycan and the proteoglycan binding motifs did not inhibit the targeting of ZG16p to ZG in AR42J cells. I have also demonstrated that when ZG16p is present in the cytoplasm it interacts with and modulates the endo-lysosomal compartment. Since it is known that impaired autophagy due to lysosomal malfunction is involved in the course of pancreatitis, a potential role of ZG16p in pancreatitis is discussed.
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Tese dout., Aquacultura, Universidade do Algarve, 2008
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Dissertação de mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015
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Viele Studienanfänger haben in der Schule wissenschaftliches Arbeiten nicht gelernt (Kunz, 1986). In der Schule unterrichtet man zwar Einzelfächer wie Mathematik, Deutsch, Englisch, Physik oder Kunst. Kaum ein Fachlehrer fühlt sich jedoch für das übergreifende Thema „Lernen lernen“ zuständig. Die Naturtalente unter den Schülern können lernen. Die anderen wursteln sich durch, mit mehr Anstrengung als nötig wäre – oder sie scheitern. Für das Studium genügt Durchwursteln nicht, man braucht effiziente Lese- und Lerntechniken. Dieses Buch hilft bei der Selbstorganisation des Studiums und bei der Bewältigung des Lernstoffs. Als Studierender profitiert man, weil man dasselbe Ziel mit weniger Aufwand erreicht oder man mit demselben Aufwand mehr erreicht. Als Lehrender profitiert man, weil gut organisierte Studierende besser und schneller lernen. Das Studium ist eine eigenständige Lebensphase, es ist keine bloße Weiterführung der Schullaufbahn, ebenso wenig ist das Studium lediglich eine Vorbereitung auf das spätere Berufsleben. Studere (lateinisch) bedeutet „sich ernsthaft um etwas bemühen“. Studieren ist nicht passives Aufnehmen, sondern aktives Gestalten (Spoun & Domnik, 2004), und das in erheblich größerem Umfang als es die Schule erfordert. Das Studium bietet mehr Freiräume als die Schule und erfordert daher mehr Selbstdisziplin und die Fähigkeit zur Selbststrukturierung der Lernprozesse sowie mehr Eigeninitiative (Streblow & Schiefele, 2006). Lernen ist Arbeit. Ohne Mühe und Anstrengung geht es daher nicht. Mit falschen Lerntechniken führt aber auch Anstrengung nicht ans Ziel (Metzig & Schuster, 2006). Man benötigt für Erfolg in Prüfungen beides: Anstrengungsbereitschaft und gute Lerntechniken. Zudem muss man nicht nur den Lernstoff beherrschen, sondern auch die Prüfungen bestehen. Deshalb werden Prüfungen in einem eigenen Kapitel behandelt. Dabei wird auf schriftliche und mündliche Prüfungsleistungen eingegangen. Im Abschnitt zur mündlichen Prüfung werden vor allem Hinweise zu Antworttechniken und Gesprächsführung sowie Tipps zum Umgang mit Nervosität und Stress vor und in der Prüfung gegeben. Die Hinweise zur Erstellung von Studienarbeiten helfen bei der ersten eigenen Arbeiten, von der Themenwahl über die Recherche bis hin zu Gliederung und formalen Vorschriften. Des Weiteren werden Klausurprüfungen behandelt, von der Vorbereitung bis zu vermeidbaren Fehlern. Dazu gibt es bisher nur wenig einschlägige Literatur. Die Hinweise in Kapitel 4 gehen deshalb zu einem Großteil auf Gespräche mit anderen Dozenten der FH Bund zurück. Sie berücksichtigen zum Teil die spezifischen Bedingungen dieser Fachhochschule, sind zum größeren Teil aber allgemein auf alle Hochschulen anwendbar. Insgesamt ist dieses Buch ein Leitfaden, der hilft, die Anforderungen an Hochschulen zu bewältigen und das Studium erfolgreich zu meistern.
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Viele Studienanfänger haben in der Schule wissenschaftliches Arbeiten nicht gelernt (Kunz, 1986). In der Schule unterrichtet man Einzelfächer wie Mathematik, Deutsch, Englisch, Physik oder Kunst. Kaum ein Fachlehrer fühlt sich jedoch für das übergreifende Thema „Lernen lernen“ zuständig. Die Naturtalente unter den Schülern können lernen. Die anderen wursteln sich durch, mit mehr Anstrengung als nötig wäre – oder sie scheitern. Für das Studium genügt Durchwursteln nicht, man braucht effiziente Arbeits- und Lerntechniken. Dieses Buch hilft bei der Selbstorganisation des Studiums und bei der Bewältigung des Lernstoffs. Als Studierender profitiert man, weil man dasselbe Ziel mit weniger Aufwand erreicht oder man mit demselben Aufwand mehr erreicht. Als Lehrender profitiert man, weil gut organisierte Studierende besser und schneller lernen. Das Studium ist eine eigenständige Lebensphase, ist keine bloße Weiterführung der Schullaufbahn. Ebenso wenig ist das Studium lediglich eine Vorbereitung auf das spätere Berufsleben. Studere (lateinisch) bedeutet „sich ernsthaft um etwas bemühen“. Studieren ist nicht passives Aufnehmen, sondern aktives Gestalten und Arbeiten (Spoun & Domnik, 2004), und das in erheblich größerem Umfang als es die Schule erfordert. Das Studium bietet mehr Freiräume als die Schule und erfordert deshalb mehr Selbstdisziplin und die Fähigkeit zur Selbststrukturierung der Lernprozesse sowie mehr Eigeninitiative (Streblow & Schiefele, 2006). Aus diesem Grund fällt vielen Studierenden das Umsteigen von der Schule oder aus dem erlernten Beruf heraus in ein Studium zunächst schwer. Lernen ist Arbeit. Ohne eigene Anstrengung geht es daher nicht. Mit falschen Lerntechniken führt aber auch Anstrengung nicht ans Ziel (Metzig & Schuster, 2006). Man benötigt für den Studienerfolg beides: Anstrengung und Lerntechniken Man muss nicht nur den Lernstoff beherrschen, sondern auch Prüfungen bestehen. In Kapitel 4 wird auf schriftliche und mündliche Prüfungsleistungen sowie auf Studienarbeiten eingegangen. Schriftliche Prüfungen werden ausführlich behandelt, von der Vorbereitung bis zu vermeidbaren Fehlern. Hierzu gibt es bisher nur wenige konkrete Hinweise in der einschlägigen Literatur. Die Ausführungen gehen deshalb zu einem Großteil auf Gespräche mit Dozentenkollegen zurück. Sie berücksichtigen zum Teil die spezifischen Bedingungen der FH Bund, sind zum größeren Teil aber allgemein auf alle Hochschulen anwendbar. Im Abschnitt zu mündlichen Prüfungen werden Hinweise zu Antworttechniken und Gesprächsführung sowie Tipps zum Umgang mit Nervosität und Stress vor und in der Prüfung gegeben. Die Hinweise zur Erstellung von Studienarbeiten helfen bei den ersten eigenen Arbeiten, von der Themenwahl über die Recherche bis hin zu Gliederung und zu formalen Vorschriften.
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Einige Kunststoffe wie die Massenkunststoffe Polypropylen (PP) und Polyethylen (PE) aber auch Polyoximethylen (POM) und Polytetrafluorethylen (PTFE) weisen eine schlechte Klebeignung auf. Werden diese Werkstoffe ohne Vorbehandlung geklebt, so können nur geringe Klebfestigkeiten erzielt werden. Aber auch bei der Herstellung von Bauteilen aus gut klebbaren Kunststoffen, wie z.B. ABS, kann eine Oberfläche entstehen, die die Klebbarkeit beeinträchtigt. Dies kann beispielsweise durch anhaftende Formtrennmittel geschehen. Auch in diesen Fällen wird beim Kleben ohne Vorbehandlung nicht die maximal mögliche Klebfestigkeit erreicht. In den aufgeführten Fällen kann jedoch durch eine geeignete Klebflächenbehandlung, in Verbindung mit einem entsprechenden Klebstoff, die Festigkeit so gesteigert werden, dass bei einer Prüfung die Verbindung im Kunststoffteil versagt.