Kvetiapinmisbruk - myte eller realitet?

publishedVersio

Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma

<p>Abstract</p> <p>Background</p> <p>Neuroblastoma is a childhood cancer derived from immature cells of the sympathetic nervous system. The disease is clinically heterogeneous, ranging from neuronal differentiated benign ganglioneuromas to aggressive metastatic tumours with poor prognosis. Amplification of the MYCN oncogene is a well established poor prognostic factor found in up to 40% of high risk neuroblastomas.</p> <p>Using neuroblastoma cell lines to study neuronal differentiation <it>in vitro </it>is now well established. Several protocols, including exposure to various agents and growth factors, will differentiate neuroblastoma cell lines into neuron-like cells. These cells are characterized by a neuronal morphology with long extensively branched neurites and expression of several neurospecific markers.</p> <p>Results</p> <p>In this study we use retrovirally delivered inducible short-hairpin RNA (shRNA) modules to knock down <it>MYCN </it>expression in <it>MYCN</it>-amplified (MNA) neuroblastoma cell lines. By addition of the inducer doxycycline, we show that the Kelly and SK-N-BE(2) neuroblastoma cell lines efficiently differentiate into neuron-like cells with an extensive network of neurites. These cells are further characterized by increased expression of the neuronal differentiation markers <it>NFL </it>and <it>GAP43</it>. In addition, we show that induced expression of retrovirally delivered anti-<it>MYCN </it>shRNA inhibits cell proliferation by increasing the fraction of MNA neuroblastoma cells in the G1 phase of the cell cycle and that the clonogenic growth potential of these cells was also dramatically reduced.</p> <p>Conclusion</p> <p>We have developed an efficient <it>MYCN</it>-knockdown <it>in vitro </it>model system to study neuronal differentiation in MNA neuroblastomas.</p

Total sitting time and risk of myocardial infarction, coronary heart disease and all-cause mortality in a prospective cohort of Danish adults

BACKGROUND: Evidence suggests that sitting time is adversely associated with health risks. However, previous epidemiological studies have mainly addressed mortality whereas little is known of the risk of coronary heart disease. This study aimed to investigate total sitting time and risk of myocardial infarction, coronary heart disease incidence and all-cause mortality. METHODS: In the Danish Health Examination Survey (DANHES) conducted in 2007-2008 we tested the hypothesis that a higher amount of daily total sitting time is associated with greater risk of myocardial infarction, coronary heart disease and all-cause mortality. The study population consisted of 71,363 men and women aged 18-99 years without coronary heart disease. Participants were followed for myocardial infarction, coronary heart disease and mortality in national registers to August 10, 2012. Cox regression analyses were performed with adjustment for potential confounders and multiple imputation for missing values. RESULTS: During a mean follow-up period of 5.4 years 358 incident cases of myocardial infarction, 1,446 of coronary heart disease, and 1,074 deaths from all causes were registered. The hazard ratios associated with 10 or more hours of daily sitting compared to less than 6 hours were 1.38 (95% CI: 1.01, 1.88) for myocardial infarction, 1.07 (95% CI: 0.91, 1.27) for coronary heart disease and 1.31 (95% CI: 1.09, 1.57). Compared to sitting less than 6 hours per day and being physically active in leisure time, the hazard ratios of sitting more than 10 hours per day and also being physically inactive in leisure time were 1.80 (95% CI: 1.15, 2.82) for myocardial infarction, 1.42 (95% CI: 1.11, 1.81) for coronary heart disease, and 2.29 (95% CI: 1.82, 2.89) for all-cause mortality. CONCLUSIONS: The results suggest that a higher amount of daily total sitting time is associated with all-cause mortality, particularly among inactive adults. In relation to coronary heart, disease results were less clear. This paper adds new evidence to the limited data on the evidence of sitting time and cardiovascular disease and mortality

Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma

BACKGROUND: MicroRNAs (miRNAs) regulate expression of many cancer-related genes through posttranscriptional repression of their mRNAs. In this study we investigate the proto-oncogene MYCN as a target for miRNA regulation. METHODS: A luciferase reporter assay was used to investigate software-predicted miRNA target sites in the 30 -untranslated region (30 UTR) of MYCN. The miRNAs were overexpressed in cell lines by transfection of miRNA mimics or miRNA-expressing plasmids. Mutation of the target sites was used to validate MYCN 30 UTR as a direct target of several miRNAs. To measure miRNA-mediated suppression of endogenous N-myc protein, inhibition of proliferation and inhibition of clonogenic growth, miRNAs were overexpressed in a MYCN-amplified neuroblastoma cell line. RESULTS: The results from this study show that MYCN is targeted by several miRNAs. In addition to the previously shown mir-34a/c, we experimentally validate mir-449, mir-19a/b, mir-29a/b/c, mir-101 and let-7e/mir-202 as direct MYCN-targeting miRNAs. These miRNAs were able to suppress endogenous N-myc protein in a MYCN-amplified neuroblastoma cell line. The let-7e and mir-202 were strong negative regulators of MYCN expression. The mir-101 and the let-7 family miRNAs let-7e and mir-202 inhibited proliferation and clonogenic growth when overexpressed in Kelly cells. CONCLUSION: The tumour-suppressor miRNAs let-7 and mir-101 target MYCN and inhibit proliferation and clonogenic growth of MYCN-amplified neuroblastoma cells

Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma

Background: Neuroblastoma is a childhood cancer derived from immature cells of the sympathetic nervous system. The disease is clinically heterogeneous, ranging from neuronal differentiated benign ganglioneuromas to aggressive metastatic tumours with poor prognosis. Amplification of the MYCN oncogene is a well established poor prognostic factor found in up to 40% of high risk neuroblastomas. Using neuroblastoma cell lines to study neuronal differentiation in vitro is now well established. Several protocols, including exposure to various agents and growth factors, will differentiate neuroblastoma cell lines into neuron-like cells. These cells are characterized by a neuronal morphology with long extensively branched neurites and expression of several neurospecific markers. Results: In this study we use retrovirally delivered inducible short-hairpin RNA (shRNA) modules to knock down MYCN expression in MYCN-amplified (MNA) neuroblastoma cell lines. By addition of the inducer doxycycline, we show that the Kelly and SK-N-BE(2) neuroblastoma cell lines efficiently differentiate into neuron-like cells with an extensive network of neurites. These cells are further characterized by increased expression of the neuronal differentiation markers NFL and GAP43. In addition, we show that induced expression of retrovirally delivered anti- MYCN shRNA inhibits cell proliferation by increasing the fraction of MNA neuroblastoma cells in the G1 phase of the cell cycle and that the clonogenic growth potential of these cells was also dramatically reduced. Conclusion: We have developed an efficient MYCN-knockdown in vitro model system to study neuronal differentiation in MNA neuroblastomas

Fitness and health benefits of team handball training for young untrained women - A cross-disciplinary RCT on physiological adaptations and motivational aspects

Purpose: The present study evaluated the effects of regular participation in small-sided team handball training on body composition, osteogenic response, physical performance, and cardiovascular risk factors, as well as well-being and motivation, in young untrained women. Methods: Twenty-eight untrained 20- to 30-year-old women were randomized to a handball training group (HG; n = 14, height 170 ± 5 cm, weight 73 ± 11 kg, VO2peak 37.7 ± 4.1 mL/min/kg) that trained 1.7 ± 0.3 times per week over 12 weeks (70 min 4 v 4 handball sessions) or an inactive control group (CG; n = 14, 169 ± 5 cm, 71 ± 12 kg, 38.1 ± 3.7 mL/min/kg). Physiological and psychological and motivational training adaptations were assessed pre- and post-intervention by dual-energy X-ray Absorptiometry (DXA) scans, blood sampling, physical tests, and questionnaires. Results: The average heart rate (HR) over all training sessions was equal to 85% ± 6% HRmax. Between-group intervention effects were observed in favor of HG for muscle mass (2.1%, p = 0.024), proximal femur bone mineral density (0.8%, p = 0.041), Yo-Yo IE1 intermittent endurance test level 1 (IE1) performance (35%, p < 0.001), and incremental treadmill test performance (11.5%, p = 0.003), but not total fat mass (p = 0.176), mean arterial blood pressure (p = 0.328), resting HR (p = 0.219), or blood lipids (p = 0.298–0.854). In CG, no changes were observed in any of the measured physiological variables after the training period. Compared to CG, HG had an increase in intrinsic motivation (p < 0.001) and in the well-being subscale “energy” (p = 0.010). Conclusion: Participation in regular recreational team handball training organized as small-sided games has marked beneficial effects on physical performance, musculoskeletal fitness, well-being, and motivation in untrained young women. Keywords: Bone mineral density (BMD), Intensity, Intermittent, Motivation, Muscle mass, Physical performance, Recreational handball, Well-bein