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Colonisation de la viande par Escherichia coli O157∶H7 : caractérisation moléculaire, cellulaire et tissulaire des interactions / Meat colonisation by Escherichia coli O157∶H7 : molecular, cellular and tissue characterisation of the interactionsChagnot, Caroline 02 April 2014 (has links)
Escherichia coli O157:H7 est le sérotype le plus souvent incriminé lors de toxi-infection alimentaire par les E. coli entérohémorragiques (EHEC). Il peut être associé, dans les cas les plus graves, à des colites hémorragiques mortelles et au syndrome hémolytique et urémique (SHU), touchant essentiellement les jeunes enfants. Le vecteur alimentaire le plus courant lors de ces contaminations est le boeuf haché. L’étape primaire de la contamination bactérienne se situe lors de l'abattage où les bactéries peuvent être transférées de la peau à la carcasse. Une gaine conjonctive entoure les muscles, sa composition protéique, similaire à la matrice extracellulaire (ECM), pourrait jouer un rôle dans l'adhésion bactérienne. Dans un premier temps, l’étude de l'adhésion et de la colonisation des bactéries aux protéines majeures de l’ECM musculaire, a révélé une forte influence des conditions de croissances sur l’adhésion, l'adhésion étant maximale à 25°C et pH7, en particulier aux collagènes I et III. Chez les EHEC, diverses protéines de surfaces peuvent être potentiellement impliquées dans l’adhésion à l’ECM. Le rôle d'un autotransporteur, l'antigène 43 (Ag43), dans l'autoagrégation, l'adhésion et la formation de biofilm, a été établit chez E. coli O157:H7 EDL933. Par la suite, les interactions entre E. coli O157:H7 et la viande ont été étudiées sur deux muscles modèles de types métabolique et contractile opposés (Soleus oxidatif lent et EDL, glycolytique rapide), caractérisés par microspectroscopie de fluorescence UV couplée au rayonnement synchrotron. Les différents types de fibres musculaires ainsi que l’effet d’une anoxie prolongée simulant la maturation des viandes ont été discriminés par leurs réponses spectrales après une excitation à 275 nm. Un tropisme bactérien plus élevé pour le muscle soleus que pour le muscle EDL a été clairement observé. Bien qu'E. coli O157:H7 adhère de manière similaire aux différents types de fibres musculaires, l'adhésion des bactéries se fait essentiellement au niveau de l'ECM, mettant en évidence le rôle clé de l'ECM et du tissu conjonctif musculaire dans l’adhésion des E. coli O157:H7 à la viande. Ces travaux de recherche sur l’adhésion bactérienne aux muscles squelettiques aux niveaux moléculaires, cellulaires et tissulaires fournissent les premières connaissances sur la physiologie des EHEC lors de la contamination de la viande et constituent un pré-requis indispensable au développement de pratiques et de stratégies innovantes afin de réduire le risque de contamination des viandes. / Escherichia coli O157:H7 is the most prevalent serotype involved in foodborne infection by enterohemorrhagic E. coli (EHEC). It is associated with life-threatening hemorrhagic colitis and the hemolyticuremic syndrome (HUS), which essentially affect young children. The major food vector of EHEC contamination is ground beef. The primary bacterial contamination occurs during the slaughter, essentially at dehiding stage where bacteria can be transferred from hides to carcasses. The connective tissue surrounding the muscle, highly similar to extracellular matrix (ECM) could potentially be a support for bacterial adhesion. When investigating the adhesion and colonization to the main muscle fibrous ECM proteins, the great influence of growth conditions on subsequent bacterial attachment was shown. Maximal adhesion to ECM proteins occurred at 25°C and pH 7, especially to collagens I and III. In EHEC, various surface-exposed protein determinants can be expressed and potentially involved in ECM adhesion. Investigating the autoaggregation, bacterial adhesion and biofilm formation, the involvement of Antigen 43 (Ag43), an autotransporter protein, was demonstrated in E. coli O157:H7 EDL933. Then, the attachment of E. coli O157:H7 to the meat was determined on two different model muscles, with different contractile and metabolic characteristic (Soleus oxidative, slow and EDL glycolytic, fast), previously characterized by UV microspectroscopy coupled to synchrotron radiation fluorescence. The different of muscle fiber types and the effect of a prolonged anoxia simulating maturing meat were discriminated by their spectral responses after excitation at 275 nm. It clearly appeared that bacteria displayed differential tropism as function of the muscle types, higher for the Soleus than the EDL muscles. While E. coli O157:H7 adhered similarly to the different types of muscle fibers, bacterial adherence essentially occurred at the ECM, pinpointing the key role of connective tissue for E. coli O157:H7 adhesion to meat. This first comprehensive investigation of bacterial adhesion to skeletal muscles at molecular, cellular and tissue levels provides new insight in the physiology of the colonization of meat by EHEC and constitutes a prerequisite for the development of innovative practices and strategies to minimize the risk of meat contamination.
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Sulfated hyaluronan alters fibronectin matrix assembly and promotes osteogenic differentiation of human bone marrow stromal cellsVogel, Sarah, Arnoldini, Simon, Möller, Stephanie, Hempel, Ute, Schnabelrauch, Matthias 28 March 2017 (has links)
Extracellular matrix (ECM) composition and structural integrity is one of many factors that influence cellular differentiation. Fibronectin (FN) which is in many tissues the most abundant ECM protein forms a unique fibrillary network. FN homes several binding sites for sulfated glycosaminoglycans (sGAG), such as heparin (Hep), which was previously shown to influence FN conformation and protein binding. Synthetically sulfated hyaluronan derivatives (sHA) can serve as model molecules with a well characterized sulfation pattern to study sGAG-FN interaction. Here is shown that the low-sulfated sHA (sHA1) interacts with FN and influences fibril assembly. The interaction of FN fibrils with sHA1 and Hep, but not with non-sulfated HA was visualized by immunofluorescent co-staining. FRET analysis of FN confirmed the presence of more extended fibrils in human bone marrow stromal cells (hBMSC)-derived ECM in response to sHA1 and Hep. Although both sHA1 and Hep affected FN conformation, exclusively sHA1 increased FN protein level and led to thinner fibrils. Further, only sHA1 had a pro-osteogenic effect and enhanced the activity of tissue non-specific alkaline phosphatase. We hypothesize that the sHA1-triggered change in FN assembly influences the entire ECM network and could be the underlying mechanism for the pro-osteogenic effect of sHA1 on hBMSC.
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Biomedicínské aplikace polykaprolaktonových nanovlákenných membrán / Biomedical applications of polycaprolactone nanofibrous matsDvořák, Pavel January 2021 (has links)
The diploma thesis deals with the treatment of polycaprolactone (PCL) nanofibers. PCL fibers were subjected to the deposition of plasma amine polymers in a low pressure pulsed radiofrequency capacitively coupled discharge using cyclopropylamine monomer (CPA). Collagen as an extracellular matrix (ECM) protein was immobilized and cell proliferation on the modified nanofiber surface was monitored. Untreated PCL fibers were also subjected to the deposition of an antibacterial copper layer, and the fibers were characterized by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and energy dispersive spectroscopy (EDX).
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Imputation of Missing Data with Application to Commodity Futures / Imputation av saknad data med tillämpning på råvaruterminerÖstlund, Simon January 2016 (has links)
In recent years additional requirements have been imposed on financial institutions, including Central Counterparty clearing houses (CCPs), as an attempt to assess quantitative measures of their exposure to different types of risk. One of these requirements results in a need to perform stress tests to check the resilience in case of a stressed market/crisis. However, financial markets develop over time and this leads to a situation where some instruments traded today are not present at the chosen date because they were introduced after the considered historical event. Based on current routines, the main goal of this thesis is to provide a more sophisticated method to impute (fill in) historical missing data as a preparatory work in the context of stress testing. The models considered in this paper include two methods currently regarded as state-of-the-art techniques, based on maximum likelihood estimation (MLE) and multiple imputation (MI), together with a third alternative approach involving copulas. The different methods are applied on historical return data of commodity futures contracts from the Nordic energy market. By using conventional error metrics, and out-of-sample log-likelihood, the conclusion is that it is very hard (in general) to distinguish the performance of each method, or draw any conclusion about how good the models are in comparison to each other. Even if the Student’s t-distribution seems (in general) to be a more adequate assumption regarding the data compared to the normal distribution, all the models are showing quite poor performance. However, by analysing the conditional distributions more thoroughly, and evaluating how well each model performs by extracting certain quantile values, the performance of each method is increased significantly. By comparing the different models (when imputing more extreme quantile values) it can be concluded that all methods produce satisfying results, even if the g-copula and t-copula models seems to be more robust than the respective linear models. / På senare år har ytterligare krav införts för finansiella institut (t.ex. Clearinghus) i ett försök att fastställa kvantitativa mått på deras exponering mot olika typer av risker. Ett av dessa krav innebär att utföra stresstester för att uppskatta motståndskraften under stressade marknader/kriser. Dock förändras finansiella marknader över tiden vilket leder till att vissa instrument som handlas idag inte fanns under den dåvarande perioden, eftersom de introducerades vid ett senare tillfälle. Baserat på nuvarande rutiner så är målet med detta arbete att tillhandahålla en mer sofistikerad metod för imputation (ifyllnad) av historisk data som ett förberedande arbete i utförandet av stresstester. I denna rapport implementeras två modeller som betraktas som de bäst presterande metoderna idag, baserade på maximum likelihood estimering (MLE) och multiple imputation (MI), samt en tredje alternativ metod som involverar copulas. Modellerna tillämpas på historisk data förterminskontrakt från den nordiska energimarkanden. Genom att använda väl etablerade mätmetoder för att skatta noggrannheten förrespektive modell, är det väldigt svårt (generellt) att särskilja prestandan för varje metod, eller att dra några slutsatser om hur bra varje modell är i jämförelse med varandra. även om Students t-fördelningen verkar (generellt) vara ett mer adekvat antagande rörande datan i jämförelse med normalfördelningen, så visar alla modeller ganska svag prestanda vid en första anblick. Däremot, genom att undersöka de betingade fördelningarna mer noggrant, för att se hur väl varje modell presterar genom att extrahera specifika kvantilvärden, kan varje metod förbättras markant. Genom att jämföra de olika modellerna (vid imputering av mer extrema kvantilvärden) kan slutsatsen dras att alla metoder producerar tillfredställande resultat, även om g-copula och t-copula modellerna verkar vara mer robusta än de motsvarande linjära modellerna.
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Softwareentwicklung ECM/WCM im Spannungsfeld KMUs–GroßunternehmenSchwarz, Oliver, Kowalewski, Christian 03 January 2020 (has links)
Der BIM-Standard hat in den letzten Jahren viel Bewegung in die Softwarelandschaft gebracht. Der Wunsch und die Notwendigkeit, Daten aus der EMC Landschaft mit den digitalen Daten der Planung und dem aktuellen Progress auf den Baustellen zu verbinden, stellt hier einen besonderen Motor dar. Die digitale Bauakte – nur ein Traum? Unternehmen unterschiedlichster Größe und Ausrichtung begleiten durch Ihre Tätigkeiten einen Neu- bzw. Umbau von der Idee bis zur Realisierung. Die Lösungsansätze bzw. Lösungen der Digitalisierung in den verschiedensten Ausprägungen, Tiefen und Teilbereichen existieren. Eine Lösung, die von den KMUs bis hin zu den Konzernen genutzt wird, haben wir aus dem Blickwinkel der Planer, Betreiber und Montageunternehmen im industriellen Anlagenbauumfeld (Industrie- und Produktionsanlagen aller Art) nicht gefunden. Die Lösung von inactio und ESZETT schaut aus zwei unterschiedlichen Blickwinkeln auf die Kunden. Die Abbildung 1 reißt das Spannungsfeld ein wenig an. inactio/eebos begleitet Ihre Kunden im Umfeld von onbase, SAP und digitalen Archiven. ESZETT kommt mit ihren Erfahrungen der Planungswelten, VR-Systemen und dem WCM-System bee aus der Planungs- und Montagebegleitung. Der Wunsch, mit einer kleinen Lösung zu beginnen (KMU), die bei den einzelnen Unternehmen für ihre Belange genutzt werden kann, hin zu der Vernetzung und der Möglichkeit des Einbindens dieser ‚Inseln‘ in einem Gesamtprojekt mit allen technischen Möglichkeiten und Verknüpfungen, soll kein Traum bleiben. [...]
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Test the ability of axolotl decellularized ECM scaffold to improve skin wound healing in miceAlariba, Walid 12 1900 (has links)
Le but de notre étude visait à déterminer si les matrices ECM (extracellular matrix) préparés à partir d'un modèle vertébré (Axolotl) capables de régénérer ses tissus suite à une blessure sont plus efficaces pour stimuler les réponses régénératives chez les animaux non régénérant (par exemple les mammifères). Nous avons testé la capacité de matrice ECM axolotl à améliorer la guérison des plaies cutanées dans des souris et nous les avons comparés à une matrice disponible commercialement (échafaudage Symbios PerioDerm) pour leur efficacité à favoriser la guérison des plaies. Des lésions d'excision ont été créées sur le dos de souris et les animaux ont été regroupés dans différents groupes; a-) ECM de peau axolotl décellularisée (groupe Axolotl), b-) matrice de derme acellulaire Symbios Perioderm (groupe PerioDerm), c-) grillage en titane (groupe témoin); respectivement. Les tissus des plaies ont été récoltés à des moments précis : 7 jours et 30 jours après la blessure pour évaluer la guérison des plaies. La guérison des blessures ayant reçu les différentes matrices a été comparées entre elles en utilisant le test de transillumination et des analyses histologiques. Les résultats indiquent que la ECM de peau d’axolotl décellularisée est bien tolérée par les souris, car aucun rejet n'a été observé. Le groupe qui a reçu l'ECM de la peau axolotl décellularisé a démontré une réépithélialisation, une densité cellulaire, une teneur en collagène (avec une organisation similaire à un tissu intact) et une vascularisation (angiogenèse) élevées par rapport aux groupes PerioDerm et témoins. La présence de follicules pileux était également observé dans le groupe axolotl (qui n'est pas présent dans PerioDerm et groupes de contrôle). Sur la base de nos résultats, l'hypothèse de base semble être correcte en ce qu'une matrice ECM provenant d'un régénérateur puissant semble favoriser la guérison plus efficacement chez les animaux normalement non régénérants. Cependant, des recherches supplémentaires devront être menées pour confirmer ces résultats. / The aim of our study sought to determine whether ECM scaffolds prepared from a vertebrate model (Axolotl) capable of regenerating tissues following injury are more effective at stimulating regenerative responses in non-regenerating animals (e.g., mammals). We tested the ability of axolotl decellularized ECM scaffolds to improve skin wound healing in mammalian models and compare the axolotl skin ECM scaffold to a commercially available one (Symbios PerioDerm scaffold) for efficiency in promoting wound healing. Excisional lesions were created on the back of mice, and animals in different groups were treated by; a-) decellularized axolotl skin ECM (Axolotl group), b-) Symbios Perioderm acellular dermis scaffold (PerioDerm group), d-) Titanized mesh only (Control group); respectively. Wound tissues were harvested at time points: 7- and 30-days post-wounding to assess the scaffolds impact on wound healing. Wound healing was compared between the Axolotl, PerioDerm and Control groups using transillumination test and histological analyses, Results indicate that the decellularized axolotl skin ECM is well tolerated by mammalian models, as no immune rejection was observed. The axolotl group that received the decellularized Axolotl Skin ECM demonstrated high reepithelialization, cellular density, collagen content (in a porous pattern similar to intact skin), vascularization (angiogenesis) compared to PerioDerm and control groups. The presence of hair follicles was also observed in the axolotl group (which is not present in PerioDerm and control groups). Based on our results, the basic hypothesis appears to be correct in that an ECM scaffold from a strong regenerator seems to promote healing more efficiently in non-regenerating animals. However, further research should be conducted to confirm these findings.
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Extracellular Matrix Synthesis and Remodeling by Mesenchymal Stromal Cells Is Context-SensitiveBurk, Janina, Sassmann, Anna, Kasper, Cornelia, Nimptsch, Ariane, Schubert, Susanna 16 January 2024 (has links)
Matrix remodeling could be an important mode of action of multipotent mesenchymal
stromal cells (MSC) in extracellular matrix (ECM) disease, but knowledge is limited in this respect.
As MSC are well-known to adapt their behavior to their environment, we aimed to investigate
if their mode of action would change in response to healthy versus pathologically altered ECM.
Human MSC-derived ECM was produced under different culture conditions, including standard
culture, culture on Matrigel-coated dishes, and stimulation with the pro-fibrotic transforming growth
factor-1 (TGF1). The MSC-ECM was decellularized, characterized by histochemistry, and used
as MSC culture substrate reflecting different ECM conditions. MSC were cultured on the different
ECM substrates or in control conditions for 2 days. Culture on ECM increased the presence of surface
molecules with ECM receptor function in the MSC, demonstrating an interaction between MSC
and ECM. In MSC cultured on Matrigel-ECM and TGF1-ECM, which displayed a fibrosis-like
morphology, gene expression of collagens and decorin, as well as total matrix metalloproteinase
(MMP) activity in the supernatant were decreased as compared with control conditions. These
results demonstrated that MSC adapt to their ECM environment, which may include pathological
adaptations that could compromise therapeutic efficacy.
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Energisimulering av effektiviseringsåtgärder vid punkthusen i Östra Sätra : Validering av byggnadsmodell samt analys av förändrade lastkurvorMorales, Leonel, Sandfors, Mats January 2016 (has links)
Det globala energibehovet ökar ständigt och i takt med detta ökar även faran för global uppvärmning bortom en hanterbar nivå och därför stramas kraven åt gällande minskade utsläpp av växthusgaser. EU har som följd av detta enats om en uppsättning klimatmål som benämns 20/20/20-målen som syftar till att minska växthusgasutsläppen med 20 procent, jämfört med 1990 års nivåer, sänka energianvändningen med 20 procent, höja andelen förnybar energi samt höja andelen biobränsle i transportsektorn med 10 procent. Sektorn bostäder och service står enligt energimyndigheten för nära 40 procent av Sveriges totala energianvändning och blir därför ett område som är mycket intressant när det gäller att implementera energieffektiviseringsåtgärder. Det kommunala företaget AB Gavlegårdarna ska under perioden 2016-2018 renovera ett tiotal punkthus i stadsdelen Östra Sätra i Gävle. Husen är typiska miljonprogramshus och beroende på val av effektiviseringsåtgärder kommer behovet av köpt värme samt el att förändras, dvs deras lastprofiler på det energisystemet. Dessa förändringar kommer i föreliggande arbete att simuleras och studeras i simuleringsprogrammet IDA ICE (IDA Indoor Climate and Energy) version 4.6.2, där resultaten jämförs och valideras med befintlig statistik. Husen som berörs av upprustningen är av typen punkthus som definieras som friliggande hus med trapphus i mitten med lägenheter runt om. Punkthusen var vanliga på 1940 talet och byggdes då oftast med 4-5 våningar. Även om huskropparna står fritt är de oftast placerade i grupp för att skapa en områdeskänsla. De aktuella byggnaderna är i stort lika varandra där avsaknad av källarplan hos vissa av husen är den tydligaste skillnaden, i övrigt består konstruktionen av markplan plus 4 våningsplan och ett vindsutrymme. Hus med källarplan rymmer totalt 29 lägenheter medan de övriga rymmer 27. Energisystemmodellen validerades samt modifierades att passa de renoverade husen. Resultatet har sedan använts för att beräkna områdets lastprofil gällande fjärrvärme vilket kan ses i nedanstående figur. Diagrammet visar lastkurvan både före och efter de energieffektiviserande åtgärderna implementerats. Med hjälp utav de erhållna effektkurvorna kan områdets totala energianvändning jämföras före och efter implementering av energieffektiviseringsåtgärder. Resultatet visar att värmebehovet sjunker med 1,39 GWh. Appliceras detta på kraftvärmeproduktionen resulterar minskningen i 0,55 GWh lägre elproduktion som måste importeras till systemet. Importeras elen från kolkondens kan detta ge upphov till ökade koldioxidutsläpp med 512 ton koldioxidekvivalenter per år jämfört med 2,13 ton CO2 ekvivalenter om elen tas från svensk elmix.
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Molekulární mechanizmy fenotypových přechodů fibroblastických buněk: dediferenciace myofibroblastů a ovlivnění invazivity a metastazování sarkomu / Molecular mechanisms of fibroblastoid cell phenotype transitions:dedifferentiation of myofibroblasts and influencing of invasiveness and metastasis of sarcomaKosla, Jan January 2013 (has links)
Fibroblasts are the principal cellular component of the connective tissue. They are a heterogeneous group of cells which contribute to the structure of connective tissue and wound healing by their ability to produce extracellular matrix (ECM). Fibroblasts and cells derived from them are involved in many pathological processes such as formation of malignant tumors and fibrosis. Tumor progression which finally leads to metastasis is a serious biomedical problem. There is a growing body of the recent evidence showing an important role of the tumor stroma and its interaction with cancer cells in cancer progression. Tumor stroma comprises mainly of myofibroblasts and their products, namely ECM, soluble factors, and enzymes. Myofibroblasts contribute more or less to all steps of cancer progression. Furthermore myofibroblasts play a key role in fibrosis, another serious human disease which is not efficiently treatable and which is associated with cancer progression. These facts made us to search for molecular means capable of eliminating the myofibroblastic phenotype. We succeeded to entirely dedifferentiate primary myofibroblasts by concomitant inhibition of TGFβ signaling and perturbation of MAPK signaling in a chick model that we have introduced. Malignant fibroblasts form sarcomas. ECM is the first...
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Knowledge technologies process and cultures : improving information and knowledge sharing at the Amateur Swimming Association (ASA)Onojeharho, Ejovwoke January 2015 (has links)
Over the last few years the ASA determined KM as a priority to assist with reducing knowledge loss, realising information assets and reducing work duplication by attempting to implement IKM tools and strategies. This research employed a pragmatic viewpoint, using a mix of both quantitative and qualitative methods to check reliability, to ensure validity while undertaking the task of implementing the IKM tools. Using a case study strategy and action research was justified, as to be pragmatic the researcher needed to understand the extent of the problem within a specified context. The research discussed in this thesis, provides a new framework for implementing KM tools; focusing on the NSO category, which the case study organisation falls into. The literature agrees enlisting influential members onto the project is vital for success; however, the findings suggested that success was not only tied to this buy-in alone, but also to the organisation s ability to retain these members for the duration of the project. The research proposed the use of a newly developed tool within the new framework, as an approach to reduce the time it takes to undertake traditional social network analysis of the organisation, as it became clear that there was a need for a method of producing updated results of the SNA, which would span the length of long projects within organisations with significantly high staff turn-over rates. Privacy was given as a factor to consider the in literature; however, the findings from this study indicated that a majority of the participants were comfortable with the system. Email knowledge extraction, and email social network systems are not new concepts, however this research presents EKESNA; a novel tool that combines both concepts in a way that allows for the continuous discovery, visualisation, and analysis of knowledge networks around specified topics of interest within an organisation; linking conversations to specific expert knowledge. EKESNA s continuous discovery of the organisation s knowledge network affords members up-to-date data to inform business process reengineering. This is a potentially ground breaking new tool that has the possibility of transforming the KM landscape in NSOs as well as a whole range of other kinds of enterprises.
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