Identification of the initial molecular changes in response to circulating angiogenic cells-mediated therapy in critical limb ischemia

BackgroundCritical limb ischemia (CLI) constitutes the most aggressive form of peripheral arterial occlusive disease, characterized by the blockade of arteries supplying blood to the lower extremities, significantly diminishing oxygen and nutrient supply. CLI patients usually undergo amputation of fingers, feet, or extremities, with a high risk of mortality due to associated comorbidities.Circulating angiogenic cells (CACs), also known as early endothelial progenitor cells, constitute promising candidates for cell therapy in CLI due to their assigned vascular regenerative properties. Preclinical and clinical assays with CACs have shown promising results. A better understanding of how these cells participate in vascular regeneration would significantly help to potentiate their role in revascularization.Herein, we analyzed the initial molecular mechanisms triggered by human CACs after being administered to a murine model of CLI, in order to understand how these cells promote angiogenesis within the ischemic tissues.MethodsBalb-c nude mice (n:24) were distributed in four different groups: healthy controls (C, n:4), shams (SH, n:4), and ischemic mice (after femoral ligation) that received either 50 mu l physiological serum (SC, n:8) or 5x10(5) human CACs (SE, n:8). Ischemic mice were sacrificed on days 2 and 4 (n:4/group/day), and immunohistochemistry assays and qPCR amplification of Alu-human-specific sequences were carried out for cell detection and vascular density measurements. Additionally, a label-free MS-based quantitative approach was performed to identify protein changes related.ResultsAdministration of CACs induced in the ischemic tissues an increase in the number of blood vessels as well as the diameter size compared to ischemic, non-treated mice, although the number of CACs decreased within time. The initial protein changes taking place in response to ischemia and more importantly, right after administration of CACs to CLI mice, are shown.ConclusionsOur results indicate that CACs migrate to the injured area; moreover, they trigger protein changes correlated with cell migration, cell death, angiogenesis, and arteriogenesis in the host. These changes indicate that CACs promote from the beginning an increase in the number of vessels as well as the development of an appropriate vascular network.Institute of Health Carlos III, ISCIII; Junta de Andaluci

Iron Behaving Badly: Inappropriate Iron Chelation as a Major Contributor to the Aetiology of Vascular and Other Progressive Inflammatory and Degenerative Diseases

The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular "reactive oxygen species" (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation). The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible. This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, etc...Comment: 159 pages, including 9 Figs and 2184 reference

Tapiolan tapa toimia autoliikekumppaniverkostossa

Tapiola-ryhmän autoliikepalveluiden sekä korvauspalveluiden ostotoiminnan toimintamallit ovat erillään toisistaan vaikka asiointi tapahtuu samojen toimijoiden kanssa. Ulkoinen asioimistapa, toimijasta riippumatta, on kaikille samanlaista eikä kohderyhmää vastaavaa. Tutkimuksen tavoitteena on ymmärtää autoliikkeiden tavoitteita, toiveita ja arkielämän ongelmia sekä selvittää miten vakuutusten myynti linkittyy näihin. Näiden tietojen pohjalta rakennetaan sisäinen ja ulkoinen toimintamalli autoliikekanavan myynti- ja ostotoiminnalle. Sisällöstä kootaan kumppanuus – käsikirja, joka on kohdennettu Tapiolan kumppanitoimintaa ajatellen. Kumppanuustoiminta on tavoitteena aloittaa vuoden 2010 aikana sekä allekirjoittaa kumppanuussopimukset 20 Suomen suurimman autoliikeketjun kanssa. Tutkimusotteena käytetään toimintatutkimusta, joka sisältää kvalitatiivisen autoliikekanavatutkimuksen, kvantitatiiviset lähtö- ja nykytilanne tyytyväisyystutkimukset, toimintamalliprosessin suunnittelu- ja rakennusvaiheen sekä toteutus- ja kehitysvaiheen seurannan ja raportoinnin. Tutkimus ajoittuu vuosille 2009-2011. Tutkimus osoittaa, että vakuutusten myynti autoliikkeissä on haasteellista. Autoliikkeet toimivat yhteistyössä monen eri toimijan kanssa, jotka osaltaan asettavat tavoitteita ja vaatimuksia liikkeille. Kovista haasteista riippumatta Tapiolan kanssa tehtyä yhteistyötä arvostetaan. Yhteyshenkilöiden toimintaa pidetään ammattitaitoisena mutta aktiivisuutta kaivataan. Tutkimus osoittaa myös sen, että verkostoitumisstrategiassa sekä sisäisen ja ulkoisen toimintamallin rakentamisessa ja toteutuksessa onnistuttiin. Vaikka suuria tyytyväisyyseroja ei kumppaneiden ja ei-kumppaneiden välille syntynyt, osoittaa tutkimus, että aktiivisuutta, panostusta ja myyjien työn tukea lisäämällä saadaan kumppanuus - yhteistyötä tiivistettyä.Tapiola-groups car dealers service and claims services purchase functions operation models are apart from each other even though business transactions happen with the same companies. The external contacts, whatever the partie, were performed the same way and not targeted to the correspondent. The aim of the research is to understand the car dealers goals, wishes and everyday problems and how insurance sales is linked to these. Based on the gathered information a partnership-guide is made for internal and external use for both the sales- and purchase functions. The partnership-guide is aimed to help the new partnership operation witch starts during the year 2010 and to help sign partnership contracts with the 20 biggest car dealerships in Finland. The research approach uses action research. The research includes a qualitative car dealership channel research, a quantitative satisfaction survey taken both prior to and after launch, designing and building the operation process and monitoring and reporting the execution and development. The research is scheduled to take place in 2009-2011. The research shows that selling insurances in car dealers has its challenges. Goals and demands are set by several different parties that car dealers work with. In spite of tough competition all parties appreciate the cooperation with Tapiola. The contact persons work professionally but more activity is desired. The research also shows that building and executing the internal and external operations model and networkingstrategy was successful. Even though there were no big differences between partnerships and no-partnerships, activity, investing and supporting the sales persons work helps to deepen the cooperation

Maggie's Embodiment of the Roma Stereotype in The Mill on the Floss

This essay focuses on Maggie in The Mill on the Floss, by George Eliot. An examination of her life is presented which is anchored in feminist critical theory and focuses on the ordeal Maggie has to endure in a patriarchal society. Furthermore, the life of the Roma is examined through postcolonial theory and compared to Maggie’s. Many of the stereotypes that emerged about the Roma are also present in Maggie’s life. It is argued that Maggie embodies the stereotypes of the Roma through her encounters with different characters in the novel.

"Dansens språk - mer som poesi än prosa" : En kvalitativ undersökning om gymnasielärares uppfattningar kring bedömning i dans

Syftet med denna studie är att undersöka gymnasielärares uppfattningar kring bedömning i dans; vad de ser som möjligheter och svårigheter. Tre kvalitativa intervjuer genomfördes och analyserades genom en fenomenografisk teoriansats. I resultatet framgår att lärarna försöker integrera två olika synsätt; det ena representerar den professionella dansvärlden där fokus ligger på känsla, intuition och produkten (föreställningen). Den andra representerar den skolastiska världen, där konkreta och sakliga betygskriterier tillsammans med processen (utvecklingen) är prioriterat. Lärarna visar alla prov på frustration över betygskriterierna, speciellt då de ska beskriva olika kvaliteter inom dansen. Dessa anses svåröversatta, och de kräver dessutom en gradering, där det klart framgår vad som skiljer olika betyg emellan. Det lärarna ser som ett tecken på lärande hos eleverna är att de har en insikt i sin egen lärprocess. Lärarna menade att det är först när eleven förstår sin egen progression som den kan gå till nästa steg och vidareutvecklas. I resultatet framgår att lärarna i hög grad använder sig av formativ bedömning i undervisningen, eftersom dans är ett praktiskt ämne. En av lärarna använder sig uteslutande av formativ bedömning och baserar betygen på sin intuition, den tysta kunskapen. Denna lärare använder sig inte av underlag eller summativ bedömning, utan processen ses som vital. De övriga två lärarna använder sig både av summativ och formativ bedömning där ett brett underlag är stöd i bedömningen och produkten och processen betonas likvärdigt i undervisningen.

The road from atherosclerosis to myocardial infarction : Studies in an experimental model

Atherosclerosis is a slow progressing disease with continuous lipid deposition and inflammation in the arterial wall. Stenotic arteries may cause angina pectoris but it is the sudden formation of an occluding thrombosis on the atherosclerotic plaque that leads to myocardial infarction. In a majority of cases such complications derive from plaque rupture or plaque erosion. Several steps in the series of events leading from silent atherosclerosis to myocardial infarction have remained unclear, in part due the lack of suitable animal models. This thesis has aimed to develop a mouse model in which myocardial infarction is triggered in order to study the physiological and molecular mechanisms that are important in the development of myocardial infarction. Isoflurane anesthetized apoE -/- x LDLR -/- and C57BL/6J mice were exposed to systemic hypoxia by reducing the inhaled oxygen concentration to 10% for 10 minutes. Mental stress was induced in conscious mice by blowing pressurized air into the cage. Physiological parameters were recorded every 30 minutes for 2-6 days by implanted transmitters. To investigate whether thrombosis was involved a thrombin inhibitor was administered in a bolus dose before the stress and after the hypoxic stress by continuous administration by osmotic mini-pumps. Controls were given PBS. Infarction development was determined by troponin T analysis and histology. The plaques and hearts were isolated at different time points after hypoxia and analysed by transcriptional profiling and quantitative real time RT-PCR. On selected genes involved in plaque vulnerability, protein expression and activity was assessed by immunohistochemistry, western blot, zymography. The results show that myocardial infarction can be induced by hypoxic stress in a two-phase pathway: an initial phase comprising a transient ischemic response which triggers a delayed second phase of ischemia and myocardial infarction. Previous results show that the first phase is endothelin dependent and in our present studies we show that the first ischemic phase triggers a delayed phase which involves thrombin. Furthermore our data show that hypoxia induces expression of inflammatory, proteolytic and pro-coagulant genes that may promote a vulnerable and pro-thrombotic plaque environment and we hypothesise that these factors may lead to endothelial or plaque damage and an activation of haemostasis. This model should be a useful tool to investigate plaque activation, thrombus formation and infarction development