CDKN2BAS is associated with periodontitis in different European populations and is activated by bacterial infection

Epidemiological studies have indicated a relationship between coronary heart disease (CHD) and periodontitis. Recently, CDKN2BAS was reported as a shared genetic risk factor of CHD and aggressive periodontitis (AgP), but the causative variant has remained unknown. To identify and validate risk variants in different European populations, we first explored 150 kb of the genetic region of CDKN2BAS including the adjacent genes CDKN2A and CDKN2B, covering 51 tagging single nucleotide polymorphisms (tagSNPs) in AgP and chronic periodontitis (CP) in individuals of Dutch origin (n=313). In a second step, we tested the significant SNP associations in an independent AgP and CP population of German origin (n=1264). For the tagSNPs rs1360590, rs3217992, and rs518394, we could validate the associations with AgP before and after adjustment for the covariates smoking, gender and diabetes, with SNP rs3217992 being the most significant (OR 1.48, 95% CI 1.19 to 1.85; p=0.0004). We further showed in vivo gene expression of CDKN2BAS, CDKN2A, CDKN2B, and CDK4 in healthy and inflamed gingival epithelium (GE) and connective tissue (CT), and detected a significantly higher expression of CDKN2BAS in healthy CT compared to GE (p=0.004). After 24 h of stimulation with Porphyromonas gingivalis in Streptococcus gordonii pre-treated gingival fibroblast (HGF) and cultured gingival epithelial cells (GECs), we observed a 25-fold and fourfold increase of CDKN2BAS gene expression in HGFs (p=0.003) and GECs (p=0.004), respectively. Considering the global importance of CDKN2BAS in the disease risk of CHD, this observation supports the theory of inflammatory components in the disease physiology of CHD

Análise de timol em cera de abelha por micro-extracção em fase sólida (SPME)

A aplicação contínua de acaricídas lipofílicos sintéticos no tratamento das abelhas conduz a uma acumulação que depende da frequência, lipofilicidade e quantidade de princípio activo utilizada. Este efeito é mais acentuado na cera de abelha que no mel, no entanto, e porque a persistência destes resíduos é elevada, provoca o aparecimento de resistências e a perda do seu efeito acaricida.[1] Esta razão levou à pesquisa de outros compostos alternativos não tóxicos e não persistentes, com efeito sobre o ácaro das abelhas, Varroa Jacobsoni. Entre estes compostos encontra-se o timol, um composto fenólico, volátil, presente no tomilho. Dos diversos componentes dos óleos essenciais este é sem dúvida o que demonstrou maior efeito acaricida, utilizando-se no tratamento das abelhas directamente ou como componente de diversas formulações.[2] Em Portugal, foi introduzido muito recentemente sob a forma comercial de APIGUARD: um gel, à base de timol, que controla termicamente a libertação do princípio activo. O controlo dos resíduos de timol na cera de abelha e no mel é assim um desafio actual quer do ponto de vista sanitário quer de qualidade alimentar. A micro-extracção em fase sólida (SPME) é uma técnica de preparação de amostras que se baseia na sorção de analítos no revestimento de uma fibra de sílica fundida e posterior desorção térmica no injector de um cromatógrafo em fase gasosa (GC). Para além de combinar num único processo etapas de extracção, purificação e concentração dos analitos, a técnica de SPME apresenta uma série de vantagens relativamente às técnicas de extracção convencionais, como a extracção líquido-líquido e extracção em fase sólida, nomeadamente a sua relativa simplicidade e rapidez, reduzido custo e não utilização de solventes para a extracção de analitos, para além de permitir a extracção por imersão directa na amostra gasosa ou líquida e extracção por amostragem do espaço-de-cabeça da amostra líquida ou sólida.[3] Ao contrário das técnicas tradicionais, que permitem uma extracção quantitativa dos analitos, a técnica de SPME baseia-se num equilíbrio de partição do analito. Esta particularidade torna a técnica de SPME bastante sensível a parâmetros experimentais que possam afectar os coeficientes de partição dos analitos e, consequentemente, a sensibilidade e reprodutibilidade dos resultados.[4] O objectivo deste trabalho é o desenvolvimento de uma metodologia para a análise de timol em ceras contaminadas, utilizando como padrão interno a benzofenona. Em primeiro lugar, procedeu-se à optimização da técnica através da determinação da quantidade de cera, temperatura de análise e período de contacto da fibra com o espaço-de-cabeça da amostra mais adequados para o caso em estudo. Numa segunda fase, procedeu-se à análise de diversas lâminas de cera contaminadas propositadamente com timol e sujeitas a diferentes condições de armazenamento: em frio, ao ar e em estufa. Finalmente, procedeu-se à construção da curva de calibração e quantificação do timol presente nas diversas amostras de cera analisadas. Considerando-se os resultados, para os níveis de contaminação avaliados, as condições analíticas mais adequadas ocorrem com a utilização de 1 g de cera, mantendo-se a fibra em contacto com o espaço-de-cabeça durante 40 minutos a uma temperatura de 60 ºC. Nestas condições experimentais foi possível obter uma boa correlação linear (r2=0,990) no intervalo de concentrações [3,5-14 mg/g]. A quantidade de timol encontrada nas amostras é significativamente inferior à colocada durante o processo de fabrico das lâminas, pelo que o processo de conservação não é o mais adequado, sendo evidente uma menor quantidade de timol quando a lâmina de cera é colocada na estufa

Проектирование автономного быстровозводимого защитного гидротехнического сооружения для местности с периодическими затоплениями

В данной выпускной квалификационной работе был проведен анализ паводковой ситуации за 20 лет. Рассмотрены инженерно-технические и организационные мероприятия по защите населения от наводнения. Исследованы современные типы быстровозводимых водоналивных дамб. Разработан проект быстровозводимого гидротехнического сооружения.In this final qualification work the analysis of a flood situation for 20 years was carried out. Technical and organizational actions for protection of the population against a flood are considered. The modern types of pre-fabricated water filling dams are investigated. The project of the pre-fabricated hydraulic engineering construction is develope

CDKN2BAS is associated with periodontitis in different European populations and is activated by bacterial infection

Epidemiological studies have indicated a relationship between coronary heart disease (CHD) and periodontitis. Recently, CDKN2BAS was reported as a shared genetic risk factor of CHD and aggressive periodontitis (AgP), but the causative variant has remained unknown. To identify and validate risk variants in different European populations, we first explored 150 kb of the genetic region of CDKN2BAS including the adjacent genes CDKN2A and CDKN2B, covering 51 tagging single nucleotide polymorphisms (tagSNPs) in AgP and chronic periodontitis (CP) in individuals of Dutch origin (n=313). In a second step, we tested the significant SNP associations in an independent AgP and CP population of German origin (n=1264). For the tagSNPs rs1360590, rs3217992, and rs518394, we could validate the associations with AgP before and after adjustment for the covariates smoking, gender and diabetes, with SNP rs3217992 being the most significant (OR 1.48, 95% CI 1.19 to 1.85; p=0.0004). We further showed in vivo gene expression of CDKN2BAS, CDKN2A, CDKN2B, and CDK4 in healthy and inflamed gingival epithelium (GE) and connective tissue (CT), and detected a significantly higher expression of CDKN2BAS in healthy CT compared to GE (p=0.004). After 24 h of stimulation with Porphyromonas gingivalis in Streptococcus gordonii pre-treated gingival fibroblast (HGF) and cultured gingival epithelial cells (GECs), we observed a 25-fold and fourfold increase of CDKN2BAS gene expression in HGFs (p=0.003) and GECs (p=0.004), respectively. Considering the global importance of CDKN2BAS in the disease risk of CHD, this observation supports the theory of inflammatory components in the disease physiology of CHD

In-lecture learning motivation predicts students’ motivation, intention, and behaviour for after-lecture learning: Examining the trans-contextual model across universities from UK, China, and Pakistan

This paper presents a cross-cultural examination of the trans-contextual model in University education setting. The purpose of the study was to test the effect of students’ perceived autonomy support and in-lecture learning motivation on motivation, intention, and behaviour with respect to after-lecture learning via the mediation of the social cognitive variables: attitude, subjective norm, and perceived behavioural control. University students from UK, China, and Pakistan completed the questionnaires of the study variables. Results revealed that in-lecture perceived autonomy support and autonomous motivation were positively associated with autonomous motivation and intention to engage in after-lecture learning activities via the mediation of the social cognitive variables in all samples. After controlling for the effect of past behaviour, relations between intention and behaviour were only observed in the Chinese sample. In conclusion, the trans-contextual model can be applied to University education, but cultural differences appear to moderate the predictive power of the model, particularly for the intention-behaviour relationship

Skin tumor formation in human papillomavirus 8 transgenic mice is associated with a deregulation of oncogenic miRNAs and their tumor suppressive targets

Background: Dysregulation of microRNA(miRNA) expression is regularly found in various types of cancer and contributes to tumorigenic processes. However, little is known about miRNA expression in non-melanoma skin cancer in which a pathogenic role of beta human papillomaviruses (HPV) is discussed. A carcinogenic potential of beta HPV8 could be demonstrated in a transgenic mouse model, expressing all early genes of HPV8 (HPV8-CER). A single UVA/B-dose induced oncogene expression and led to papilloma growth within three weeks. Objective: Expression of miRNAs and their targets during HPV8-mediated tumor formation in mice. Methods: Skin of untreated or UV-irradiated wild-type and HPV8-CER mice was analyzed for miRNA expression and localization by qPCR and in situ hybridization. MiRNA target protein expression was analyzed by immunohistochemical staining. Results: Early steps in skin tumor formation in HPV8-CER mice were associated with an upregulation of the oncogenic miRNA-17-5p, -21 and -106a and a downregulation of the tumor-suppressive miRNA-155 and -206, which could be demonstrated by qPCR and in situ hybridization. The respective targets of miRNA-21 and -106a, the tumor suppressors PTEN, PDCD4 and Rb with their pivotal role in cell cycle regulation, apoptosis and proliferation were found to be downregulated. Conclusion: This is the first report demonstrating that a cutaneous HPV type deregulates the expression of miRNAs. These deregulations are closely related to the UV-induced upregulation of HPV8 oncogene levels, which suggest a direct or indirect HPV8-specific effect on miRNA expression. These data presume that HPV8 interferes with the miRNA mediated gene regulation to induce tumorigenesis. (C) 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved

Ligand-Induced Stabilization of the Native Human Superoxide Dismutase 1

A common characteristic of familial (fALS) and sporadic amyotrophic lateral sclerosis (sALS) is the accumulation of aberrant proteinaceous species in the motor neurons and spinal cord of ALS patients—including aggregates of the human superoxide dismutase 1 (hSOD1). hSOD1 is an enzyme that occurs as a stable dimeric protein with several post-translational modifications such as the formation of an intramolecular disulfide bond and the acquisition of metal cofactors that are essential for enzyme activity and further contribute to protein stability. Some mutations and/or destabilizing factors promote hSOD1 misfolding, causing neuronal death. Aggregates containing misfolded wild-type hSOD1 have been found in the spinal cords of sALS as well as in non-hSOD1 fALS patients, leading to the hypothesis that hSOD1 misfolding is a common part of the ALS pathomechanism. Therefore, stabilizing the native conformation of SOD1 may be a promising approach to prevent the formation of toxic hSOD1 species and thus ALS pathogenesis. Here, we present the 16-mer peptide S1VL-21 that interferes with hSOD1 aggregation. S1VL-21 was identified by phage display selection with the native conformation of hSOD1 as a target. Several methods such as microscale thermophoresis (MST) measurements, aggregation assays, and cell viability assays revealed that S1VL-21 has a micromolar binding affinity to native hSOD1 and considerably reduces the formation of hSOD1 aggregates. This present work therefore provides the first important data on a potential lead compound for hSOD1-related drug development for ALS therapy