pDEST FG12-CMV DsRed Vector

Melanoma is the most rapidly increasing malignancy among young people in the United States. If detected early, the disease is easily treated; however, once the disease has metastasized it is largely refractory to conventional therapies and is associated with a high mortality rate. The development of human cancer from a pre-malignant primary tumor to a metastatic lesion that develops at secondary sites is thought to be a multi-step process, requiring many genetic and epigenetic events that provide a growth advantage to cells. It is still unclear which of the many genetic changes in human cancers are required for metastasis. Therefore, it is critical to evaluate each step in the metastatic process. To this end, we will generate novel lentiviral vectors containing fluorescent reporter genes to better understand the metastatic potential of melanoma cells. Vectors containing green fluorescent protein (GFP) have already been generated while vectors containing red fluorescent protein (RFP) and yellow fluorescent protein (YFP) will be cloned. Viruses will be generated and used to infect syngeneic explanted tumor cells. Since each vector will be marked with a reporter gene of a different color, we will be able to track the movement of these cells in vivo and determine the source of each metastatic tumor. Whole body fluorescence will be detected using the FluorVivo Imaging System (INDEC BioSystems, Santa Clara, CA). The experiments proposed will contribute to an increased understanding of the biology of melanoma, which has the potential to identify specific molecular targets and promote the development of more effective therapies for advanced stages of this disease

Characterizing and inhibiting two pathways activated in Glioblastoma Multiforme

Despite major improvements in imaging, radiation, and surgery, the prognosis for patients with Glioblastoma multiforme (GBM) remains clinically challenging. New treatment strategies are badly needed to reduce the mortality and morbidity associated with this disease. The resistance of these tumors to conventional treatments makes GBM patients ideal candidates for molecularly targeted therapies and several agents are currently being developed(1). Because GBM is genetically heterogeneous, combination therapies or the use of multikinase inhibitors are more likely to achieve the greatest therapeutic benefit(2,3). However, genes that can be productively targeted for effective therapies in patients remain to be identified. The overall objective of this project was to better understand the signaling pathways driving cell survival so that new targets can be identified in gliomas. These studies will lead to an increased understanding of the proteins that are altered in this disease and should provide promising opportunities to develop better treatment strategies based on specific molecular targets. Two parallel pathways, which are both activated in GBM, converge on downstream survival signaling cascades. Studies have demonstrated that blocking only one pathway often leads to a transient response (e.g., delayed time to progression), but tumors eventually progress(4). More effective therapies are likely to be those that inhibit more than one target or pathway(5). Targeting antiapoptotic Bcl-2 proteins in combination with RAS/MAPK or AKT/mTOR inhibition is a rationale approach. To determine if inhibiting both the RAS/MAPK and AKT/mTOR pathways in combination results in increased apoptosis in glioma cells, I compared the level of apoptosis in cells treated with each inhibitor alone and in combination. Treatment of glioma cells with a MEK inhibitor in combination with a PI(3)K inhibitor has not previously been reported and therefore represents a new approach in the field. We already know that just inhibiting RAS/MAPK or AKT/mTOR alone results in cell cycle arrest but not death. I tested the effect on cell death when combining the inhibitors of both pathways, and saw an increase in cell death. I determined the growth inhibitory and apoptotic sensitivity of several human glioma cell lines to inhibition of both RAS/MAPK and AKT/mTOR pathways. Due to the heterogeneous nature of GBM, I predicted and saw that these cell lines display varying levels of sensitivity to MEK/PI(3)K inhibition. These differences can then be used in the future to further define the mechanism(s) by which the AKT and MAPK pathways mediate survival signaling in glioma cells

Defining the role of NRAS in melanoma maintenance

The incidence of melanoma has increased 600 percent over the last four decades; it is the most rapidly increasing malignancy among young people in the United States and is currently the leading cause of cancer death in women aged 25- 29. If detected early, the disease is easily treated; however, once the disease has metastasized it is largely refractory to conventional therapies and is associated with a high mortality rate. The development of cancer from a pre-malignant primary tumor to a metastatic cancer that develops at secondary sites is a multi-step process, thought to require many genetic and epigenetic events that provide a growth advantage to cells. It is still unclear, however, which of the many genetic changes are required late in tumor progression. The increased incidence of melanoma, combined with the poor prognosis of patients with advanced disease, make it imperative that we increase our understanding of the underlying genetic causes of melanoma such that better targeted therapeutic strategies can be developed

Life style related to blood pressure and body weight in adolescence: Cross sectional data from the Young-HUNT study, Norway

Background The associations between physical activity, unhealthy dietary habits and cigarette smoking and blood pressure, overweight and obesity are well established in adulthood. This is not the case for similar associations in adolescence. Thus, the purpose of this study is to examine how physical activity, smoking status and dietary habits were related to overweight, obesity and blood pressure in a population of Norwegian adolescents. Methods Weight, height, systolic (SBP) and diastolic blood pressure (DBP) were measured, and body mass index (BMI) was calculated among 8408 adolescents who participated in a population based study in 1995–97 in the county of Nord-Trøndelag. Internationally accepted cut-off values were used to determine if the adolescents were overweight or obese. The adolescents also completed a detailed questionnaire including dietary habits, physical activity and smoking habits. We calculated adjusted mean blood pressures and odds ratios for being overweight or obese for different exposure categories of life style variables. Results Low levels of physically activity were associated with increased odds of being overweight (odds ratio (OR), 1.4; 95% confidence interval (CI), 1.1–1.8 in girls and OR, 2.0; 95% CI, 1.6–2.5 in boys) or obese (girls: OR, 3.1; 95% CI, 1.6–6.0; boys: OR, 3.7; 95% CI, 2.1–6.4). In addition, the least physically active girls had a 1.5 mmHg higher mean DBP compared with the most active (p-trend <0.001), and among boys this difference was 1.0 mmHg (p-trend < 0.001). Smokers were more likely to be obese (OR, 1.6; 95% CI, 1.1–2.5 in girls and 1.4; 95% CI, 0.9–2.1 in boys) compared with non-smokers. Smokers also had lower mean SBP than non-smokers; however, this finding was restricted to smokers with the lowest smoking exposure. Associations between dietary habits and weight status largely disappeared after adjusting for weight losing behaviour. Conclusion In this population of adolescents low levels of physical activity were associated with higher mean DBP and higher odds of overweight or obesity. Smoking was also associated with higher odds of overweight and obesity. The paradoxical associations between healthy dietary habits and overweight and obesity are most likely an effect of reverse causality

Reading and writing difficulties in adolescence and later risk of welfare dependence. A ten year follow-up, the HUNT Study, Norway

Background: Welfare dependence and low work participation among young people have raised concern in many European countries. Reading and writing difficulties (RWD) might make young people vulnerable to work integration problems and welfare dependence through negative influences on education and health. Our main objective of this study was to examine if RWD in adolescence affected the risk of welfare dependence in young adulthood. Methods: Baseline information on self-reported RWD, health and family was obtained for 8950 school-attending adolescents in Nord-Trøndelag County, Norway, participating in the Young-HUNT1 survey, 1995-97. All individuals were linked to biological parents to identify siblings and parental education from national registers. Welfare dependence was assessed by the reception of social benefits (medical and economic) from the national social insurance database (1998-2007). Only long-term benefits (> 180 days) were included. Results: The adolescents who reported RWD at baseline were more likely to receive medical or social benefits during follow-up compared with those who did not report RWD. In girls with RWD, the adjusted 5-year risk (at age 24 to 28) for receiving medical benefits was 0.20 (95% confidence interval 0.14-0.26), compared with 0.11 (0.09- 0.12) in girls without RWD. In boys the corresponding risks were 0.13 (0.09-0.17) and 0.08 (0.07-0.09). Conclusions: The associations between RWD in adolescence and welfare dependence later in life suggest that increased attention should be paid to these problems when discussing the public health aspects of work integration, since there might be a potential for prevention

Alcohol intoxication and mental health among adolescents--a population review of 8983 young people, 13-19 years in North-Trondelag, Norway: the Young-HUNT Study

BACKGROUND: The aims of this study were to describe alcohol use among Norwegian teenagers and investigate the associations between mental health problems and alcohol intoxications with focus on age and gender. METHODS: Population based, cross-sectional survey addressing all adolescents aged 13-19 years, attending secondary or high school in North - Trondelag County, Norway. 8983 youths (91%) answered the Young-HUNT questionnaire in the 1995-1997 survey. Logistic regression models were used to study associations.RESULTS: 80% of the respondents reported that they had tried drinking alcohol, and 57% had been intoxicated at least once. The proportion of the students, which had tried alcohol, was equal in both genders and increased with age. Attention problems and conduct problems were strongly associated with frequent alcohol intoxications in both genders. Anxiety and depressive symptoms among girls were also related to high numbers of intoxications CONCLUSION: Gender differences in number of alcohol intoxications were small. There was a close association between both conduct and attention problems and high alcohol consumption in both genders. Girls with symptoms of anxiety and depression reported more frequent alcohol intoxications

How do relationships begin?

In this paper we address the issue ‘How do relationships begin?’ Based on a review of work within theIMP Approach on stage and state models of relationship evolution, we conclude that very littleattention has been paid to beginnings of relationships. We discuss why this might be so, and why theissue deserves more consideration. Based on a case study, we make a first start at discussing how wemay conceptualise and discuss relationship beginnings. Furthermore, we depict a firm’s ‘relationshipinitiation profile’ and suggest that a firm may benefit from examining its profile and the costs andbenefits associated with it. Lastly, we propose issues which may be pursued in further research.Keywords: relationship evolution, stage model, state model, beginning, relationship initiation profile

No association between chronic musculoskeletal complaints and Val158Met polymorphism in the Catechol-O-methyltransferase gene. The HUNT study

BACKGROUND: The Catechol-O-methyltransferase (COMT) gene contains a functional polymorphism, Val158Met, that has been found to influence human pain perception. In one study fibromyalgia was less likely among those with Val/Val genotype. METHODS: In the 1995–97 Nord-Trøndelag Health Study (HUNT), the association between Val/Met polymorphism at the COMT gene and chronic musculoskeletal complaints (MSCs) was evaluated in a random sample of 3017 individuals. RESULTS: The distribution of the COMT Val158Met genotypes and alleles were similar between controls and the twelve different chronic MSCs groups. Even when the Met/Met and Val/Met genotypes were pooled, the distribution of the Val/Val genotype and other genotypes were similar between controls and the chronic MSCs groups. CONCLUSION: In this population-based study, no significant association was found between Val/Met polymorphism at the COMT gene and chronic MSCs

Cerebrospinal fluid proteomics targeted for central nervous system processes in bipolar disorder

The etiopathology of bipolar disorder is largely unknown. We collected cerebrospinal fluid (CSF) samples from two independent case-control cohorts (total n = 351) to identify proteins associated with bipolar disorder. A panel of 92 proteins targeted towards central nervous system processes identified two proteins that replicated across the cohorts: the CSF concentrations of testican-1 were lower, and the CSF concentrations of C-type lectin domain family 1 member B (CLEC1B) were higher, in cases than controls. In a restricted subgroup analysis, we compared only bipolar type 1 with controls and identified two additional proteins that replicated in both cohorts: draxin and tumor necrosis factor receptor superfamily member 21 (TNFRSF21), both lower in cases than controls. This analysis additionally revealed several proteins significantly associated with bipolar type 1 in one cohort, falling just short of replicated statistical significance in the other (tenascin-R, disintegrin and metalloproteinase domain-containing protein 23, cell adhesion molecule 3, RGM domain family member B, plexin-B1, and brorin). Next, we conducted genome-wide association analyses of the case-control-associated proteins. In these analyses, we found associations with the voltage-gated calcium channel subunit CACNG4, and the lipid-droplet-associated gene PLIN5 with CSF concentrations of TNFRSF21 and CLEC1B, respectively. The reported proteins are involved in neuronal cell-cell and cell-matrix interactions, particularly in the developing brain, and in pathways of importance for lithium’s mechanism of action. In summary, we report four novel CSF protein associations with bipolar disorder that replicated in two independent case-control cohorts, shedding new light on the central nervous system processes implicated in bipolar disorder