In vitro anti-angiogenic properties of LGD1069, a selective retinoid X-receptor agonist through down-regulating Runx2 expression on Human endothelial cells

<p>Abstract</p> <p>Background</p> <p>LGD1069 (Targretin^®) is a selective retinoid X receptor (RXR) ligand, which is used in patients for cutaneous T-cell lymphoma. Our published study reported that LGD1069 inhibited tumor-induced angiogenesis in non-small cell lung cancer. In present study, we found that LGD1069 suppressed the proliferation, adhesion, invasion and migration of endothelial cells directly, and affected the expression of vegf and some matrix genes.</p> <p>Methods</p> <p>Human umbilical vein endothelial cells (HUVECs) were used for <it>in vitro </it>study. MTT assay and Sulforhodamine B assay were used for cell viability assay; the tube formation assay was used to investigate the effect of LGD1069 on angiogenesis <it>in vitro</it>. <it>In vitro </it>adhesion, migration and invasion of HUVEC cells were analyzed by Matrigel adhesion, migration and invasion assay. Gene expressions were measured by RT-PCR and Western blot analysis.</p> <p>Results</p> <p>Our data showed here that LGD1069 inhibited the activation of TGF-β/Smad pathway significantly. Furthermore, it was demonstrated that expression of Runx2 was suppressed pronouncedly during incubation with LGD1069. Runx2 is a DNA-binding transcription factor which plays a master role in tumor-induced angiogenesis and cancer cells metastasis by interaction with the TGF-β/Smad pathway of transcriptional modulators.</p> <p>Conclusions</p> <p>Our results suggested that LGD1069 may impair angiogenic and metastatic potential induced by tumor cells through suppressing expression of Runx2 directly on human endothelial cells, which may point out new pathway through which LGD1069 display anti-angiogenic properties, and provide new molecular evidence to support LGD1069 as a potent anti-metastatic agent in cancer therapy.</p

Tamoxifen and Flaxseed Alter Angiogenesis Regulators in Normal Human Breast Tissue In Vivo

The incidence of breast cancer is increasing in the Western world and there is an urgent need for studies of the mechanisms of sex steroids in order to develop novel preventive strategies. Diet modifications may be among the means for breast cancer prevention. Angiogenesis, key in tumor progression, is regulated by the balance between pro- and anti-angiogenic factors, which are controlled in the extracellular space. Sampling of these molecules at their bioactive compartment is therefore needed. The aims of this study were to explore if tamoxifen, one of the most used anti-estrogen treatments for breast cancer affected some of the most important endogenous angiogenesis regulators, vascular endothelial growth factor (VEGF), angiogenin, and endostatin in normal breast tissue in vivo and if a diet supplementation with flaxseed had similar effects as tamoxifen in the breast. Microdialysis was used for in situ sampling of extracellular proteins in normal breast tissue of women before and after six weeks of tamoxifen treatment or before and after addition of 25 g/day of ground flaxseed to the diet or in control women. We show significant correlations between estradiol and levels of VEGF, angiogenin, and endostatin in vivo, which was verified in ex vivo breast tissue culture. Moreover, tamoxifen decreased the levels of VEGF and angiogenin in the breast whereas endostatin increased significantly. Flaxseed did not alter VEGF or angiogenin levels but similar to tamoxifen the levels of endostatin increased significantly. We conclude that one of the mechanisms of tamoxifen in normal breast tissue include tipping of the angiogenic balance into an anti-angiogenic state and that flaxseed has limited effects on the pro-angiogenic factors whereas the anti-angiogenic endostatin may be modified by diet. Further studies of diet modifications for breast cancer prevention are warranted

17β-Estradiol Enhances Breast Cancer Cell Motility and Invasion via Extra-Nuclear Activation of Actin-Binding Protein Ezrin

Estrogen promotes breast cancer metastasis. However, the detailed mechanism remains largely unknown. The actin binding protein ezrin is a key component in tumor metastasis and its over-expression is positively correlated to the poor outcome of breast cancer. In this study, we investigate the effects of 17β-estradiol (E2) on the activation of ezrin and its role in estrogen-dependent breast cancer cell movement. In T47-D breast cancer cells, E2 rapidly enhances ezrin phosphorylation at Thr567 in a time- and concentration-dependent manner. The signalling cascade implicated in this action involves estrogen receptor (ER) interaction with the non-receptor tyrosine kinase c-Src, which activates the phosphatidylinositol-3 kinase/Akt pathway and the small GTPase RhoA/Rho-associated kinase (ROCK-2) complex. E2 enhances the horizontal cell migration and invasion of T47-D breast cancer cells in three-dimensional matrices, which is reversed by transfection of cells with specific ezrin siRNAs. In conclusion, E2 promotes breast cancer cell movement and invasion by the activation of ezrin. These results provide novel insights into the effects of estrogen on breast cancer progression and highlight potential targets to treat endocrine-sensitive breast cancers

The interplay of matrix metalloproteinase-8, transforming growth factor-beta 1 and vascular endothelial growth factor-C cooperatively contributes to the aggressiveness of oral tongue squamous cell carcinoma

Background: Matrix metalloproteinase-8 (MMP-8) has oncosuppressive properties in various cancers. We attempted to assess MMP-8 function in oral tongue squamous cell carcinoma (OTSCC). Methods: MMP-8 overexpressing OTSCC cells were used to study the effect of MMP-8 on proliferation, apoptosis, migration, invasion and gene and protein expression. Moreover, MMP-8 functions were assessed in the orthotopic mouse tongue cancer model and by immunohistochemistry in patient samples. Results: MMP-8 reduced the invasion and migration of OTSCC cells and decreased the expression of MMP-1, cathepsin-K and vascular endothelial growth factor-C (VEGF-C). VEGF-C was induced by transforming growth factor-beta 1 (TGF-beta 1) in control cells, but not in MMP-8 overexpressing cells. In human OTSCC samples, low MMP-8 in combination with high VEGF-C was an independent predictor of poor cancer-specific survival. TGF-beta 1 treatment also restored the migration of MMP-8 overexpressing cells to the level of control cells. In mouse tongue cancer, MMP-8 did not inhibit metastasis, possibly because it was eliminated in the peripheral carcinoma cells. Conclusions: The suppressive effects of MMP-8 in OTSCC may be mediated through interference of TGF-beta 1 and VEGF-C function and altered proteinase expression. Together, low MMP-8 and high VEGF-C expression have strong independent prognostic value in OTSCC.Peer reviewe

Human matrix metalloproteinases: An ubiquitarian class of enzymes involved in several pathological processes

Human matrix metalloproteinases (MMPs) belong to the M10 family of the MA clan of endopeptidases. They are ubiquitarian enzymes, structurally characterized by an active site where a Zn(2+) atom, coordinated by three histidines, plays the catalytic role, assisted by a glutamic acid as a general base. Various MMPs display different domain composition, which is very important for macromolecular substrates recognition. Substrate specificity is very different among MMPs, being often associated to their cellular compartmentalization and/or cellular type where they are expressed. An extensive review of the different MMPs structural and functional features is integrated with their pathological role in several types of diseases, spanning from cancer to cardiovascular diseases and to neurodegeneration. It emerges a very complex and crucial role played by these enzymes in many physiological and pathological processes

Aspects of improving and maintaining physical activity in patients with hip or knee osteoarthritis

Aim: This thesis aims to enhance knowledge of how people with osteoarthritis should be managed and supported in order to increase and maintain physical activity in the long-term.  Method: Study I and study II was based on a randomised controlled study (RCT) including 141 osteoarthritis patients. The short and long-term effect of an individualised physical activity on prescription intervention compared with individualised advice about physical activity were evaluated. The primary outcome was physical activity and secondary outcomes were fitness/performance, pain and quality of life, evaluated at 6, 12 and 24 months. In Study III, 7-day diaries were evaluated regarding which forms of physical activity i.e. walking, swimming, cycling, gardening etc. the patients chose themselves and maintained after one and two years. In addition were evaluated, which category these activities belonged to: aerobic, muscle strength, mind-body or everyday activity, and whether there were differences in characteristic of the patients in the different forms. In study IV responsiveness of function, how well instruments captured an improvement, one year after a physical activity intervention was measured. Two unilateral performance-based tests were compared with a bilateral performance-based test and with questionnaires about function.  Results: The RCT provided no evidence that individualised physical activity on prescription differ from individualised advice on physical activity in improving short and long-term physical activity, function, pain and quality of life (Study I and II). Walking was the form of physical activity performed most frequently and best maintained after 12 and 24 months. Walking were preferred by women, older individuals and individuals with weak legs while men also preferred everyday activity and cycling. Few patients preferred strength training (Study III). The maximal step-up test (one-leg testing) was more responsive to change in physical function (SMD effect size 0.57) compare to the bilateral 30-second chair-stand test (0.48) (Study IV).  Conclusion: There is still absence of evidence for any particular physical activity intervention to effectively increase physical activity in the long-term in osteoarthritis patients. Individual counselling with support to choose preferred physical activities that are easy to perform in daily life may be a beneficial approach for long-term maintenance

Airwasher : Luftrening med hjälp av växter

Det här examensarbetet omfattar en designprocess innehållande, litteraturstudier, intervjuer, observationer, marknadsundersökning, idégenerering, fokusgrupp, utvärdering och kommunikation. Syftet var att vidareutveckla en luftrenare/luftfuktare kallad Airwasher för företaget Växt och Miljödesign. Växt och Miljödesign är ett litet företag som grundar sin verksamhet på att hyra ut och underhålla växter till andra företag. De hyr även ut sin egen uppfinning Airwasher. Airwasher bygger på växters naturliga luftreningsegenskaper då den med hjälp av en fläkt suger in luft från rummet och med hjälp av ett rör för den ner till rötterna där en reningsprocess sker. Växt och Miljödesign hade inte några önskemål på hur Airwasher skulle vidareutvecklas utan det var upp till mig att hitta områden med utvecklingspotential. För att identifiera vilka problem som fanns med Airwasher gjordes intervjuer och observationer ute hos kunder. Det visade sig att det största problemet var att hitta lämpliga platser för Airwasher eftersom den endast kommer i en golvstående kruka med en diameter på 50 cm. Efter att problemen var identifierade började en idégenereringsprocess med målet att hitta lösningar. Alla lösningar som kom fram till en början skulle inte förbättra produkten nämnvärt men däremot göra den mer komplicerad. Det var inte önskvärt så jag tog ett steg tillbaka och funderade över det verkliga behovet, vilket var att förse kunderna med frisk luft. För att göra det på bästa sätt bestämde jag mig för att fokusera på att utveckla tjänsten Airwasher, inte produkten. Slutresultatet blev en serie med tre olika storlekar anpassade efter Växt och Miljödesigns sortiment av krukor. På så vis skulle man kunna utnyttja alla naturliga platser för blomkrukor och enkelt få frisk luft över hela kontoret. För att bestämma ett formspråk för nya Airwasher samlade jag en fokusgrupp där tre värdeord som Airwasher står för diskuterades fram. Orden som bestämdes var naturlig, diskret och pålitlig. Utifrån dessa följde ett skissarbete, först på papper sedan i 3-dimensionellt medium. De bästa idéerna togs ut för att sedan utvärderas mot samma fokusgrupp. När formen var bestämd valdes lämplig produktionsmetod utifrån Växt och Miljödesigns förutsättningar, vilket var 3D-skrivning. För att hålla sig till värdeorden valdes ett träfärgat filament av sågspån och PLA-plast, vilket passar bra in i krukans naturliga färgpalett

Aspects of improving and maintaining physical activity in patients with hip or knee osteoarthritis

Aim: This thesis aims to enhance knowledge of how people with osteoarthritis should be managed and supported in order to increase and maintain physical activity in the long-term.  Method: Study I and study II was based on a randomised controlled study (RCT) including 141 osteoarthritis patients. The short and long-term effect of an individualised physical activity on prescription intervention compared with individualised advice about physical activity were evaluated. The primary outcome was physical activity and secondary outcomes were fitness/performance, pain and quality of life, evaluated at 6, 12 and 24 months. In Study III, 7-day diaries were evaluated regarding which forms of physical activity i.e. walking, swimming, cycling, gardening etc. the patients chose themselves and maintained after one and two years. In addition were evaluated, which category these activities belonged to: aerobic, muscle strength, mind-body or everyday activity, and whether there were differences in characteristic of the patients in the different forms. In study IV responsiveness of function, how well instruments captured an improvement, one year after a physical activity intervention was measured. Two unilateral performance-based tests were compared with a bilateral performance-based test and with questionnaires about function.  Results: The RCT provided no evidence that individualised physical activity on prescription differ from individualised advice on physical activity in improving short and long-term physical activity, function, pain and quality of life (Study I and II). Walking was the form of physical activity performed most frequently and best maintained after 12 and 24 months. Walking were preferred by women, older individuals and individuals with weak legs while men also preferred everyday activity and cycling. Few patients preferred strength training (Study III). The maximal step-up test (one-leg testing) was more responsive to change in physical function (SMD effect size 0.57) compare to the bilateral 30-second chair-stand test (0.48) (Study IV).  Conclusion: There is still absence of evidence for any particular physical activity intervention to effectively increase physical activity in the long-term in osteoarthritis patients. Individual counselling with support to choose preferred physical activities that are easy to perform in daily life may be a beneficial approach for long-term maintenance

Stadgar vs konstnärskap – hur kan de harmonieras?

This work is a discussion between myself and a couple of different professionals in the opera industry about stage fright, the importance of preparing and the positives and negative with preparing in different ways. The essay explains the different types of auditions with focus on auditions for scholarships and how to adapt to the statutes of the scholarship without losing your artistry. It ends with a recollection of my five latest large auditions, how they went and my reflections on the result versus my preparations. They also conclude how I can work specifically moving forward since the results were vastly different with the different levels of preparation