\u3ci\u3ePTHR1/SOX9\u3c/i\u3e and \u3ci\u3eIDH1/IDH2\u3c/i\u3e Relative Expression in Primary Chondrocyte and Chondrosarcoma Cells Under the Synergistic Influence of Inducible Hypoxia and Extracellular Acidosis

Cartilage cells (Chondrocytes) grow in rather unique environmental conditions in the human body. Cartilage is avascular tissue and lacks innervation. Its main source of nutrients is derived from the synovial fluid and/or perichondrium. Consequently, these cells must survive and thrive under hypoxic and acidic stressors. Published data suggests that there are a multitude of genes affected from either one of these two stressors or both. However, these factors are frequently overlooked in cartilage research, and results are reported in either normoxia/pH=7.0 conditions, or they only account for one of the conditions. The scope of this study is to examine how these stressors affect gene expression in primary chondrocytes and chondrosarcomas. In this study, one primary chondrocyte cell line (CON5) and two chondrosarcoma grade II cell lines, JJ012-IDH1 mutant and SW1353- IDH2 mutant, were grown in four experimental conditions: hypoxia (5% O2), acidosis (pH=5.5), hypoxia and acidosis, and normoxia/(pH=7). Four genes of interest were analyzed via RT-qPCR relative to the ACTB housekeeping gene: parathyroid hormone receptor-1 (PTHR1), SRY-box transcription factor 9 (SOX9), and isocitrate dehydrogenase 1 and 2 (IDH1/IDH2). PTHR1 and SOX9 keep chondrocytes in a proliferative state and delay their hypertrophy. On the other hand, IDH1 and IDH2 are metabolic enzymes that convert isocitrate to α-ketoglutarate (α-KG). Their mutant counterparts further convert α-KG into a competitive oncometabolite D-2-Hydroxygluterate (D-2-HG). Our colorimetric assay data suggest that D-2-HG concentration levels are 10.5-fold and 6-fold more elevated in JJ012/SW1353 respectively than in the IDH wild type CON5. Our gene expression data indicates that both inducible hypoxia and extracellular acidosis alter gene expression not only separately but also when combined. This study further highlights the importance of these stressors in cartilage biology research.

Reforms of the public sector management in Albania

The problems treated and the often calumniated problems about public administration in Albania gave an impulse to our research. In inquiring through the comparative analysis of the tow public and private sector, we tried to certify the hypotheses that the public administration has enough problems, and that the reform in a certain point have positive indications, but there’s place for improvement. We could say that the paths leading to the integration of the large European family are, among others, the effective reforms in the public sector management, too.peer-reviewe

Relational Urbanism: A Framework for Variability

In a context of rapid urbanization and increasingly standardized built environments, urbanism must find new methods of creating appropriate conditions for the variability of contemporary urban life. The city, understood as a system of interconnected processes in constant change, offers a relational way of thinking about urban design. This thesis explores the concept of Relational Urbanism through a strategic design approach that engages the complexity of the site to create variability in the built environment by relating built form to landscape elements. This relational approach has particular potential in post-industrial sites, where challenging existing conditions and processes of remediation resist conventional methods of redevelopment. The thesis focuses on the Toronto Port Lands as a testing ground for this design approach, drawing on the site's industrial heritage to develop a landscape framework and a set of relational rules that will guide the emergence of a diverse urban environment able to change over time. A series of design strategies—remediation parks, urban delta, adapted industry, and differentiated fabric—rethink the challenges of the site as opportunities for public benefit, creating a variegated landscape for built form to respond to. In contrast to a singular static master plan, this method favours multiple flexible strategies that can be deployed incrementally, breaking down the scale of development and allowing it to be realized by a wide variety of stakeholders. Through this approach the thesis seeks to enable the city to intentionally but subtly guide its urban landscape toward diversity and allow its citizens to participate in its continued adaptation

A Nonprofit Model in a For-Profit World: A Closer Look at Sheltered Workshops and Sustainability as an Employee Run Business

Historically, society has tended to isolate and segregate individuals with intellectual and developmental disabilities. Despite improvements such forms of discrimination continue to be a serious social problem. On October 11, 2011, the Department of Justice began an investigation into several state’s systems of providing vocational services to individuals with intellectual and developmental disabilities. This action came about due to several states being out of compliance around Title II of the Americans with Disabilities Act (ADA). Title II of the ADA prohibits discrimination on the basis of disability for all services, programs and activities provided to the public by state and local governments. Given recent federal policy changes and directives the State of Massachusetts has created a plan developed by leaders consisting of representatives of the Association of Developmental Disabilities Providers (ADPP), The Arc of Massachusetts and the Massachusetts Department of Developmental Services (DDS) to increase opportunities for integrated employment for individuals with intellectual disabilities (ID). The intention of this plan is to phase out group employment settings otherwise known as sheltered workshops

Gene Expression Under Combined Hypoxia And Acidosis In Chondrosarcoma

Chondrosarcomas are the second most common cause of bone cancer and are removed surgically with wide margins. On recurrence, they are resistant to chemo and radiation therapy and new treatment options are critically required. This tumor type produces hyaline cartilage, a cartilage normally formed under hypoxic and acidic environment due to lack of vasculature in cartilage. Paradoxically, chondrosarcomas arise in the well vascularized, oxygen rich environment of the bone. Hypoxia and acidosis are two stressors where the cellular effects are typically reported separately even though cells experience combined effects of hypoxia and acidosis. Given the mechanistic links between hypoxia and acidosis we hypothesized that gene expression profiles will be differentially changed when chondrosarcoma cells were exposed to individual compared to combined stressors. We investigated expression of four genes expressed during cartilage and cartilage tumor formation in primary chondrocytes and two grade II chondrosarcoma cell lines, SW1353 and JJ012. Two genes, PTH1R and SOX9 are known to respond to hypoxia and acidosis separately. Two genes, IDH1 and IDH2, are mutated in chondrosarcoma cell lines JJ012 and SW1353 respectively. These mutations confer a condition of false hypoxia on the cells through stabilization of HIF-1α. The result is chondrosarcoma cells metabolize glycolytically through aerobic glycolysis. How the cells respond to hypoxia and acidosis is of considerable interest as metabolically the cells are molecularly predisposed to these conditions. Our gene expression data found that combined hypoxia and extracellular acidosis alter gene expression compared to either stressor alone. Cells showed gene specific responses to stressors that were cell type specific likely indicating influence on gene expression regulatory sequences. The importance of this work is highlighting that conditions under which cells are investigated is crucial and should be considered when measuring cell response to in vitro treatment exposures

STIMULATION AND GOOD MANAGEMENT OF FOOD PRODUCTION-A MOST IMPORTANT INDICATOR FOR A CONSTANT REGIONAL DEVELOPMENT

“The national economic development is based on the sustainable development of the regions of the country”. Thisconfirmation is visible in the paper presented. The paper is focused on development of the agricultural sector region ofKorca mainly in the traditional farming plants of the region, and the enforcing the food-processing industry. In thepaper, it is presented the present situation of food-processing industry in the region of Korca, and the further, it isgiven the situation in conservation of vegetables, as part of the a food-processing industry. This, because of the factthat Korca has had a good tradition in cultivating traditional vegetables. The papers brings new values some of whichare :• The focus on an important sector of the economic development of the district of Korce such as the agro-elaboratinindustry.• The full treatment of the present situation of this industry in the region of Korce.• The focus on the consumer demands of the region for canned vegetable products.• The picked up conclusions will serve as a firm basement for further research work in this field

The right to vote in the Albanian and italian law

L’Albania legifera in un Parlamento unicamerale composto da 140 deputati, eletti per un periodo di quattro anni. Il diritto di voto è ancora un argomento molto discusso in Albania Chiarire quello che è stato finora il viaggio e la dimensione del diritto di voto nella Repubblica dell’Albania, è un impegno importante, perché permette al popolo albanese e i suoi politici di guardare al futuro senza ripetere gli stessi errori del passato e di vedere al futuro e alla necessità di miglioramenti da una prospettiva più ampia. In questo senso, l’analisi delle disposizioni normative e costituzionali, ma anche le interpretazioni costituzionali da parte della Corte Costituzionale albanese e del Collegio elettorale che chiariscono queste disposizioni diventa una necessità. La seconda parte della tesi è dedicata al diritto di voto in Italia. Devo sottolineare che è stato particolarmente difficile per me portare qualcosa di originale dato che il diritto di voto è un argomento infinitamente trattato e per molti aspetti completamente esaurito. Questo lo dimostra una vasta gamma di letteratura e di giurisprudenza da parte della Corte costituzionale italiana presa in esame. In questo modo, ogni tentativo di riflessione finisce per apparire semplificato.Albania legislates in a unicameral parliament composed of 140 deputies, elected for a term of four years. The right to vote is still a hot topic in Albania. Clarify what has been the journey so far and the size of the voting right in the Republic of Albania, is an important commitment, because it allows the Albanian people and it’s politicians to look to the future without repeating the same mistakes of the past and see the future and the need of improvements from a broader perspective. I this sense, the analysis of laws and constitutional provisions, bur also the constitutional interpretations by the Albanian Constitutional Court and the Electoral College that clarify this provisions becomes a necessity. The second part of the thesis is dedicated to the right to vote in Italy. I must stress that it was particularly difficult for me to bring something original as the right to vote is a subject infinitely treated and in many aspects totally completed. This is demonstrated by a wide range of literature and jurisprudence by the Italian Constitutional Court examined. In this way, any attempt of reflections end up looking simplified