Lower bone strength in young patients with Fontan circulation compared to controls.

OBJECTIVES: Previous reports indicate bone deficits in patients with Fontan circulation. However, the consequences of these deficits on bone strength and when these changes occur are unclear. AIM: To compare the tibial bone strength-strain index between young patients (6-19 years) with Fontan circulation and age- and sex-matched controls, and to determine strength-strain-index in subgroups of children (6-12 years) and adolescents (13-19 years) versus controls. METHOD: The tibia was examined with peripheral quantitative CT. Based on the assessed data, bone strength-strain index was calculated in the lateral and anterior-posterior directions. RESULTS: Twenty patients with Fontan and twenty controls (mean age 13.0 ± 4.4 years; 50% females) were examined. Patients had a lower strength-strain index in the lateral direction compared to controls (808.4 ± 416.8mm3 versus 1162.5 ± 552.1mm3, p = 0.043). Subgroup analyses showed no differences regarding strength-strain index in children (6-12 years) with Fontan circulation compared to controls. However, the adolescents (13-19 years) with Fontan circulation had lower strength-strain indexes in both the lateral and anterior-posterior directions compared to controls (1041.4 ± 299.8mm3 versus 1596.4 ± 239.6mm3, p < 0.001, and 771.7 ± 192.4mm3 versus 1084.9 ± 215.0mm3, p = 0.004). When adjusted for height, there were differences between patients (6-19 years) and controls in strength-strain indexes in both the lateral and anterior-posterior directions. In subgroup analyses, the results remained robust. CONCLUSION: Young patients (6-19 years) with Fontan circulation have a lower strength-strain index in the tibia compared to controls. Subgroup analyses show that this deficit is mainly driven by the differences in adolescents (13-19 years), which might suggest that bone strength decreases with age.

Vitamin D, liver-related biomarkers, and distribution of fat and lean mass in young patients with Fontan circulation.

INTRODUCTION/AIM: Young patients with Fontan circulation may have low serum 25-hydroxyvitamin D levels, an affected liver, and unhealthy body compositions. This study aimed to explore the association between vitamin D intake/levels, liver biomarkers, and body composition in young Fontan patients. METHOD: We collected prospective data in 2017 to 2018, obtained with food-frequency questionnaires, biochemical analyses of liver biomarkers, and dual-energy X-ray absorptiometry scans in 44 children with Fontan circulation. Body compositions were compared to matched controls (n = 38). Linear regression analyses were used to investigate associations of biomarkers, leg pain, and lean mass on serum levels of 25-hydroxyvitamin D. Biomarkers were converted to z scores and differences were evaluated within the Fontan patients. RESULTS: Our Fontan patients had a daily mean vitamin D intake of 9.9 µg and a mean serum 25-hydroxyvitamin D of 56 nmol/L. These factors were not associated with fat or lean mass, leg pain, or biomarkers of liver status. The Fontan patients had significantly less lean mass, but higher fat mass than controls. Male adolescents with Fontan circulation had a greater mean abdominal fat mass than male controls and higher cholesterol levels than females with Fontan circulation. CONCLUSION: Vitamin D intake and serum levels were not associated with body composition or liver biomarkers in the Fontan group, but the Fontan group had lower lean mass and higher fat mass than controls. The more pronounced abdominal fat mass in male adolescents with Fontan circulation might increase metabolic risks later in life.

Impaired knee extension muscle strength in adolescents but not in children with Fontan circulation.

INTRODUCTION: Impaired isometric muscle strength was previously reported in adults with Fontan circulation. However, it is unclear if this impairment is present in children and adolescents with Fontan circulation. We investigated isometric muscle strength of the lower limb in patients (6-18 years) with Fontan circulation in comparison with healthy controls. METHOD: In this cross-sectional study, 43 patients (6-18 years) with Fontan circulation and 43 age- and sex-matched controls were included. Isometric knee extension and plantar flexion muscle strength were assessed using dynamometry (Newton, N). Lean mass of the legs was assessed with dual-energy X-ray absorptiometry. Analyses were performed on group level (n = 43), and for subgroups that included children aged 6-12 years (n = 18) and adolescents aged 13-18 years (n = 25). RESULTS: On group level, the patients with Fontan circulation had impaired isometric knee extension strength in comparison with the controls (p = 0.03). In subgroup analyses, impaired isometric knee extension strength was present in the adolescents (p = 0.009) but not in the children groups. For plantar flexion, there was no difference between patients and controls. There was no difference in lean mass between patients and controls (9.6 ± 4.3 kg vs. 10.8 ± 5.6 kg, p = 0.31). However, the lean mass was highly correlated to isometric knee extension strength (patients r = 0.89, controls r = 0.96, p < 0.001) and isometric plantar flexion strength (patients r = 0.7, controls r = 0.81, p < 0.001). CONCLUSION: The finding of impaired isometric knee extension muscle strength in adolescents (13-18 years) with Fontan circulation and no corresponding impairment in the children group (6-12 years) could imply that isometric muscle strength gets more impaired with age.

Fluid restriction negatively affects energy intake and growth in very low birthweight infants with haemodynamically significant patent ductus arteriosus.

AIM: We explored if fluid restriction in very low birthweight (VLBW) infants with a haemodynamically significant patent ductus arteriosus (PDA) affected energy and protein intakes and growth. METHODS: Retrospectively, we identified 90 VLBW infants that were admitted to Umea University Hospital, Sweden, between 2009 and 2012: 42 with and 48 without haemodynamically significant PDA (hsPDA). Anthropometric, fluid, energy and protein intake data during the first 28 days of life were expressed as z-scores. RESULTS: In the 42 infants diagnosed with hsPDA, fluid intake was restricted after diagnosis, resulting in a decrease in energy and protein intake. No decrease was observed in the other 48 infants in the cohort. Multivariate analysis showed that the z-score of weight change depended on both ductus arteriosus status and energy intake; thus, infants with hsPDA did not grow as expected with the energy provided to them. CONCLUSION: Energy and protein intake was diminished in prematurely born infants with hsPDA when fluid was restricted after diagnosis. The initial reduction in intakes may have contributed to the lower postnatal growth observed in these infants.

Cross-sectional associations between maternal self-efficacy and dietary intake and physical activity in four-year-old children of first-time Swedish mothers.

BACKGROUND: Healthy dietary and physical activity behaviours are established early in life where children learn by observing their parents. Therefore, parents can act as role models and influence their children toward a healthier lifestyle. Besides a strong association between parental and child health behaviours, parents also influence their children's health behaviours through socio-cognitive processes, where perceived self-efficacy is the central component. The objective was to examine if parental self-efficacy among Swedish mothers was associated with their four-year-old children's dietary and physical activity behaviours. METHODS: This cross-sectional study was based on information from control participants that took part in the Swedish primary prevention trial of childhood obesity (PRIMROSE) (n = 420 mother-child pairs). Linear regression models were used to examine the associations between parental self-efficacy (Parental Self-Efficacy for Promoting Healthy Physical Activity and Dietary Behaviours in Children Scale) and children's dietary intake (parent reported) and levels of physical activity (accelerometer) with adjustments for potential confounders. RESULTS: Mothers' efficacy beliefs in promoting healthy dietary or physical activity behaviours in their children were associated with a slightly higher consumption of fruit and vegetables among their children (ß: 0.03 [95%CI: 0.01; 0.04] P < 0.001) and slightly higher levels of moderate-to-vigorous activity (ß: 0.43 [95%CI: 0.05; 0.81] P = 0.03). Mothers' belief in their ability to limit unhealthy dietary and physical activity behaviours was inversely associated with children's intake of unhealthy snacks (ß: -0.06 [95%CI: -0.10; -0.02] P < 0.01). CONCLUSION: Our cross-sectional study suggests weak positive correlations between maternal self-efficacy and healthy dietary and physical activity behaviours, and weak inverse associations between maternal self-efficacy and unhealthy dietary and physical activity behaviours among their children.

Analysis artefacts of the INS-IGF2 fusion transcript.

BACKGROUND: In gene expression analysis, overlapping genes, splice variants, and fusion transcripts are potential sources of data analysis artefacts, depending on how the observed intensity is assigned to one, or more genes. We here exemplify this by an in-depth analysis of the INS-IGF2 fusion transcript, which has recently been reported to be among the highest expressed transcripts in human pancreatic beta cells and its protein indicated as a novel autoantigen in Type 1 Diabetes. RESULTS: Through RNA sequencing and variant specific qPCR analyses we demonstrate that the true abundance of INS-IGF2 is >20,000 fold lower than INS in human beta cells, and we suggest an explanation to the nature of the artefacts which have previously led to overestimation of the gene expression level in selected studies. We reinvestigated the previous reported findings of detection of INS-IGF2 using antibodies both in Western blotting and immunohistochemistry. We found that the one available commercial antibody (BO1P) raised against recombinant INS-IGF2 show strong cross-reaction to native proinsulin, and we did not detect INS-IGF2 protein in the human beta cell line EndoC-ßH1. Furthermore, using highly sensitive proteomics analysis we could not demonstrate INS-IGF2 protein in samples of human islets nor in EndoC-ßH1. CONCLUSIONS: Sequence features, such as fusion transcripts spanning multiple genes can lead to unexpected results in gene expression analysis, and care must be taken in generating and interpreting the results. For the specific case of INS-IGF2 we conclude that the abundance of the fusion transcript/protein is exceedingly lower than previously reported, and that current immuno-reagents available for detecting INS-IGF2 protein have a strong cross-reaction to native human proinsulin. Finally, we were unable to detect INS-IGF2 protein by proteomics analysis.

Primary prevention of childhood obesity through counselling sessions at Swedish child health centres: design, methods and baseline sample characteristics of the PRIMROSE cluster-randomised trial.

BACKGROUND: Childhood obesity is a growing concern in Sweden. Children with overweight and obesity run a high risk of becoming obese as adults, and are likely to develop comorbidities. Despite the immense demand, there is still a lack of evidence-based comprehensive prevention programmes targeting pre-school children and their families in primary health care settings. The aims are to describe the design and methodology of the PRIMROSE cluster-randomised controlled trial, assess the relative validity of a food frequency questionnaire, and describe the baseline characteristics of the eligible young children and their mothers. METHODS/DESIGN: The PRIMROSE trial targets first-time parents and their children at Swedish child health centres (CHC) in eight counties in Sweden. Randomisation is conducted at the CHC unit level. CHC nurses employed at the participating CHC received training in carrying out the intervention alongside their provision of regular services. The intervention programme, starting when the child is 8-9 months of age and ending at age 4, is based on social cognitive theory and employs motivational interviewing. Primary outcomes are children's body mass index and waist circumference at four years. Secondary outcomes are children's and mothers' eating habits (assessed by a food frequency questionnaire), and children's and mothers' physical activity (measured by accelerometer and a validated questionnaire), and mothers' body mass index and waist circumference. DISCUSSION: The on-going population-based PRIMROSE trial, which targets childhood obesity, is embedded in the regular national (routine) preventive child health services that are available free-of-charge to all young families in Sweden. Of the participants (n = 1369), 489 intervention and 550 control mothers (75.9%) responded to the validated physical activity and food frequency questionnaire at baseline (i.e., before the first intervention session, or, for children in the control group, before they reached 10 months of age). The food frequency questionnaire showed acceptable relative validity when compared with an 8-day food diary. We are not aware of any previous RCT, concerned with the primary prevention of childhood obesity through sessions at CHC that addresses healthy eating habits and physical activity in the context of a routine child health services programme. TRIAL REGISTRATION: ISRCTN16991919.

Exploring the Possible Impact of Oral Nutritional Supplements on Children's Oral Health: An In Vitro Investigation.

Eight pediatric oral nutritional supplements (ONSs) and 0.5% fat bovine milk were examined in vitro regarding their effect on the adhesion of three caries-related bacteria, Streptococcus mutans (strain CCUG 11877T), Lactobacillus gasseri (strain CCUG 31451), and Scardovia wiggsiae (strain CCUG 58090), to saliva-coated hydroxyapatite, as well as their pH and capacity to withstand pH changes. Bacteria were cultivated and radiolabeled. The adhesion assays used synthetic hydroxyapatite coated with whole or parotid saliva. Measurements of pH and titration of the products with HCl and NaOH were conducted in triplicate. Three ONSs promoted the S. mutans adhesion to saliva-coated hydroxyapatite (increase from 35% to >200%), supporting caries risk enhancement. S. wigssiae and L. gasseri adhered only to one and no ONS, respectively. Most supplements had limited buffering capacity to counteract acidification changes, suggesting their low capacity to neutralize acids, and one ONS showed a significant capacity to counteract basic changes, suggesting a high erosive potential. S. mutans adhesion was influenced by the ONS pH and volume NaOH added to reach pH 10. L. gasseri and S. wiggsiae adhesion was influenced by the ONSs' carbohydrate and fat content. Interdisciplinary efforts are needed to increase awareness and prevent the possible negative impact of ONSs on children's oral health.

Large-Scale Functional Genomics Screen to Identify Modulators of Human ß-Cell Insulin Secretion.

Type 2 diabetes (T2D) is a chronic metabolic disorder affecting almost half a billion people worldwide. Impaired function of pancreatic ß-cells is both a hallmark of T2D and an underlying factor in the pathophysiology of the disease. Understanding the cellular mechanisms regulating appropriate insulin secretion has been of long-standing interest in the scientific and clinical communities. To identify novel genes regulating insulin secretion we developed a robust arrayed siRNA screen measuring basal, glucose-stimulated, and augmented insulin secretion by EndoC-ßH1 cells, a human ß-cell line, in a 384-well plate format. We screened 521 candidate genes selected by text mining for relevance to T2D biology and identified 23 positive and 68 negative regulators of insulin secretion. Among these, we validated ghrelin receptor (GHSR), and two genes implicated in endoplasmic reticulum stress, ATF4 and HSPA5. Thus, we have demonstrated the feasibility of using EndoC-ßH1 cells for large-scale siRNA screening to identify candidate genes regulating ß-cell insulin secretion as potential novel drug targets. Furthermore, this screening format can be adapted to other disease-relevant functional endpoints to enable large-scale screening for targets regulating cellular mechanisms contributing to the progressive loss of functional ß-cell mass occurring in T2D.

General practitioners' and district nurses' conceptions of the encounter with obese patients in primary health care.

BACKGROUND: Primary health care specialists have a key role in the management of obesity. Through understanding how they conceive the encounter with patients with obesity, treatment may be improved. The aim of this study was thus to explore general practitioners' and district nurses' conceptions of encountering patients with obesity in primary health care. METHOD: Data were collected through semi-structured interviews, and analysed using a phenomenographic approach. The participants were 10 general practitioners (6 women, 4 men) and 10 district nurses (7 women, 3 men) from 19 primary health care centres within a well-defined area of Sweden. RESULTS: Five descriptive categories were identified: Adequate primary health care, Promoting lifestyle change, Need for competency, Adherence to new habits and Understanding patient attitudes. All participants, independent of gender and profession, were represented in the descriptive categories. Some profession and gender differences were, however, found in the underlying conceptions. The general staff view was that obesity had to be prioritised. However, there was also the contradictory view that obesity is not a disease and therefore not the responsibility of primary health care. Despite this, staff conceived it as important that patients were met with respect and that individual solutions were provided which could be adhered to step-by-step by the patient. Patient attitudes, such as motivation to change, evasive behaviour, too much trust in care and lack of self-confidence, were, however, conceived as major barriers to a fruitful encounter. CONCLUSIONS: Findings from this study indicate that there is a need for development and organisation of weight management in primary health care. Raising awareness of staff's negative views of patient attitudes is important since it is likely that it affects the patient-staff relationship and staff's treatment efforts. More research is also needed on gender and profession differences in this area.

Comparison of DNA Methylation Profiles of Hemostatic Genes between Liver Tissue and Peripheral Blood within Individuals.

DNA methylation has become increasingly recognized in the etiology of complex diseases, including thrombotic disorders. Blood is often collected in epidemiological studies for genotyping and has recently also been used to examine DNA methylation in epigenome-wide association studies. DNA methylation patterns are often tissue-specific, thus, peripheral blood may not accurately reflect the methylation pattern in the tissue of relevance. Here, we collected paired liver and blood samples concurrently from 27 individuals undergoing liver surgery. We performed targeted bisulfite sequencing for a set of 35 hemostatic genes primarily expressed in liver to analyze DNA methylation levels of >10,000 cytosine-phosphate-guanine (CpG) dinucleotides. We evaluated whether DNA methylation in blood could serve as a proxy for DNA methylation in liver at individual CpGs. Approximately 30% of CpGs were nonvariable and were predominantly hypo- (<25%) or hypermethylated (>70%) in both tissues. While blood can serve as a proxy for liver at these CpGs, the low variability renders these unlikely to explain phenotypic differences. We therefore focused on CpG sites with variable methylation levels in liver. The level of blood-liver tissue correlation varied widely across these variable CpGs; moderate correlations (0.5 ≤ r < 0.75) were detected for 6% and strong correlations (r ≥ 0.75) for a further 4%. Our findings indicate that it is essential to study the concordance of DNA methylation between blood and liver at individual CpGs. This paired blood-liver dataset is intended as a resource to aid interpretation of blood-based DNA methylation results.

Perceived discrimination among men and women with normal weight and obesity. A population-based study from Sweden.

AIMS: We examined whether men and women with obesity reported different types of discrimination to a greater extent than those with normal weight, and explored whether these associations were modified by socioeconomic position. METHOD: National representative sample of men and women, with normal weight (n = 2,000), moderate obesity (n = 2,461) and severe obesity (n = 557). Participants were identified in a yearly population-based survey (1996-2006) and data on perceived discrimination and potential confounding factors were measured in 2008. Logistic regression models tested whether obesity was associated with perceived lifetime, workplace, healthcare and interpersonal discrimination. RESULTS: The overall response rate was 56%. For men, moderate obesity was associated with workplace discrimination, while severely obese women were more likely to report this sort of discrimination than normal weight women. Severely obese individuals were twice as likely to report healthcare discrimination than normal weight individuals. Women, regardless of weight status group, were in turn twice as likely to report healthcare discrimination as men. Women with severe obesity were significantly more likely to report interpersonal discrimination compared with normal weight women. Socioeconomic position modified the association between weight status and healthcare discrimination. Highly educated individuals with moderate and severe obesity were more likely to report healthcare discrimination than their normal weight counterparts, whereas low educated individuals with normal weight, moderate and severe obesity were equally likely to report discrimination. CONCLUSIONS: In this large, population-based study, discrimination was more likely to be reported by obese individuals compared with those of normal weight. The associations, however, varied according to gender and socioeconomic position.

A Comprehensive Sequencing-Based Analysis of Allelic Methylation Patterns in Hemostatic Genes in Human Liver.

Characterizing the relationship between genetic, epigenetic (e.g., deoxyribonucleic acid [DNA] methylation), and transcript variation could provide insights into mechanisms regulating hemostasis and potentially identify new drug targets. Several hemostatic factors are synthesized in the liver, yet high-resolution DNA methylation data from human liver tissue is currently lacking for these genes. Single-nucleotide polymorphisms (SNPs) can influence DNA methylation in cis which can affect gene expression. This can be analyzed through allele-specific methylation (ASM) experiments. We performed targeted genomic DNA- and bisulfite-sequencing of 35 hemostatic genes in human liver samples for SNP and DNA methylation analysis, respectively, and integrated the data for ASM determination. ASM-associated SNPs (ASM-SNPs) were tested for association to gene expression in liver using in-house generated ribonucleic acid-sequencing data. We then assessed whether ASM-SNPs associated with gene expression, plasma proteins, or other traits relevant for hemostasis using publicly available data. We identified 112 candidate ASM-SNPs. Of these, 68% were associated with expression of their respective genes in human liver or in other human tissues and 54% were associated with the respective plasma protein levels, activity, or other relevant hemostatic genome-wide association study traits such as venous thromboembolism, coronary artery disease, stroke, and warfarin dose maintenance. Our study provides the first detailed map of the DNA methylation landscape and ASM analysis of hemostatic genes in human liver tissue, and suggests that methylation regulated by genetic variants in cis may provide a mechanistic link between noncoding SNPs and variation observed in circulating hemostatic proteins, prothrombotic diseases, and drug response.

An atlas of human metabolism.

Genome-scale metabolic models (GEMs) are valuable tools to study metabolism and provide a scaffold for the integrative analysis of omics data. Researchers have developed increasingly comprehensive human GEMs, but the disconnect among different model sources and versions impedes further progress. We therefore integrated and extensively curated the most recent human metabolic models to construct a consensus GEM, Human1. We demonstrated the versatility of Human1 through the generation and analysis of cell- and tissue-specific models using transcriptomic, proteomic, and kinetic data. We also present an accompanying web portal, Metabolic Atlas (https://www.metabolicatlas.org/), which facilitates further exploration and visualization of Human1 content. Human1 was created using a version-controlled, open-source model development framework to enable community-driven curation and refinement. This framework allows Human1 to be an evolving shared resource for future studies of human health and disease.

Semantic text mining in early drug discovery for type 2 diabetes.

BACKGROUND: Surveying the scientific literature is an important part of early drug discovery; and with the ever-increasing amount of biomedical publications it is imperative to focus on the most interesting articles. Here we present a project that highlights new understanding (e.g. recently discovered modes of action) and identifies potential drug targets, via a novel, data-driven text mining approach to score type 2 diabetes (T2D) relevance. We focused on monitoring trends and jumps in T2D relevance to help us be timely informed of important breakthroughs. METHODS: We extracted over 7 million n-grams from PubMed abstracts and then clustered around 240,000 linked to T2D into almost 50,000 T2D relevant 'semantic concepts'. To score papers, we weighted the concepts based on co-mentioning with core T2D proteins. A protein's T2D relevance was determined by combining the scores of the papers mentioning it in the five preceding years. Each week all proteins were ranked according to their T2D relevance. Furthermore, the historical distribution of changes in rank from one week to the next was used to calculate the significance of a change in rank by T2D relevance for each protein. RESULTS: We show that T2D relevant papers, even those not mentioning T2D explicitly, were prioritised by relevant semantic concepts. Well known T2D proteins were therefore enriched among the top scoring proteins. Our 'high jumpers' identified important past developments in the apprehension of how certain key proteins relate to T2D, indicating that our method will make us aware of future breakthroughs. In summary, this project facilitated keeping up with current T2D research by repeatedly providing short lists of potential novel targets into our early drug discovery pipeline.

Prejudice against obesity among 10-year-olds: a nationwide population-based study.

AIMS: Population-based research on children's possible prejudice against thin, obese and average body sizes is scarce. This study examined children's prejudice against various body sizes of both sexes. The effects of sex, body size, place of residence and socio-economic status (SES) on children's prejudice were also investigated. METHODS: In 2005, a nationally representative sample of 10-year-old children (N = 1409) responded to a questionnaire measuring stereotypes and prejudice against thin, average-weight and obese silhouettes. RESULTS: As estimated by odds ratios (ORs), children were more likely to report prejudice against obesity (OR = 53, 95% confidence interval [CI]: 42-67) and thinness (OR = 20, 95% CI: 16-26) than against average body weight. The risk of being prejudiced varied with child's sex and with sex and body size of the silhouette figure. No association was found between own body weight and prejudice against various body sizes. Children with high SES were more likely to be prejudiced against obesity compared with children with low SES (OR = 1.2, 95% CI: 1.1-1.4). CONCLUSION: This large, population-based study showed that Swedish 10-year-old children hold stereotypical attitudes and are prejudiced against not only peers with obesity but also those with thin body sizes. Interestingly, no association was found between own body weight and prejudice against various body sizes.

Hemostatic Genes Exhibit a High Degree of Allele-Specific Regulation in Liver.

OBJECTIVE: Elucidating the genetic basis underlying hepatic hemostatic gene expression variability may contribute to unraveling genetic factors contributing to thrombotic or bleeding disorders. We aimed to identify novel cis-regulatory variants involved in regulating hemostatic genes by analyzing allele-specific expression (ASE) in human liver samples. STUDY DESIGN: Biopsies of human liver tissue and blood were collected from adults undergoing liver surgery at the Sahlgrenska University Hospital (n = 20). Genomic deoxyribonucleic acid (gDNA) and total ribonucleic acid (RNA) were isolated. A targeted approach was used to enrich and sequence 35 hemostatic genes for single nucleotide polymorphism (SNP) analysis (gDNAseq) and construct individualized genomes for transcript alignment. The allelic ratio of transcripts from targeted RNAseq was determined via ASE analysis. Public expression quantitative trait loci (eQTL) and genome-wide association study (GWAS) data were used to assess novelty and importance of the ASE SNPs (and proxies, r 2 ≥ 0.8) for relevant traits/diseases. RESULTS: Sixty percent of the genes studied showed allelic imbalance across 53 SNPs. Of these, 7 SNPs were previously validated in liver eQTL studies. For 32 with eQTLs in other cell/tissue types, this is the first time genotype-specific expression is demonstrated in liver, and for 14 ASE SNPs, this is the first ever reported genotype-expression association. A total of 29 ASE SNPs were previously associated with the respective plasma protein levels and 17 ASE SNPs to other relevant GWAS traits including venous thromboembolism, coronary artery disease, and stroke. CONCLUSION: Our study provides a comprehensive ASE analysis of hemostatic genes and insights into the regulation of hemostatic genes in human liver.

The EndoC-ßH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates.

OBJECTIVE: To characterize the EndoC-ßH1 cell line as a model for human beta cells and evaluate its beta cell functionality, focusing on insulin secretion, proliferation, apoptosis and ER stress, with the objective to assess its potential as a screening platform for identification of novel anti-diabetic drug candidates. METHODS: EndoC-ßH1 was transplanted into mice for validation of in vivo functionality. Insulin secretion was evaluated in cells cultured as monolayer and as pseudoislets, as well as in diabetic mice. Cytokine induced apoptosis, glucolipotoxicity, and ER stress responses were assessed. Beta cell relevant mRNA and protein expression were investigated by qPCR and antibody staining. Hundreds of proteins or peptides were tested for their effect on insulin secretion and proliferation. RESULTS: Transplantation of EndoC-ßH1 cells restored normoglycemia in streptozotocin induced diabetic mice. Both in vitro and in vivo, we observed a clear insulin response to glucose, and, in vitro, we found a significant increase in insulin secretion from EndoC-ßH1 pseudoislets compared to monolayer cultures for both glucose and incretins. Apoptosis and ER stress were inducible in the cells and caspase 3/7 activity was elevated in response to cytokines, but not affected by the saturated fatty acid palmitate. By screening of various proteins and peptides, we found Bombesin (BB) receptor agonists and Pituitary Adenylate Cyclase-Activating Polypeptides (PACAP) to significantly induce insulin secretion and the proteins SerpinA6, STC1, and APOH to significantly stimulate proliferation. ER stress was readily induced by Tunicamycin and resulted in a reduction of insulin mRNA. Somatostatin (SST) was found to be expressed by 1% of the cells and manipulation of the SST receptors was found to significantly affect insulin secretion. CONCLUSIONS: Overall, the EndoC-ßH1 cells strongly resemble human islet beta cells in terms of glucose and incretin stimulated insulin secretion capabilities. The cell line has an active cytokine induced caspase 3/7 apoptotic pathway and is responsive to ER stress initiation factors. The cells' ability to proliferate can be further increased by already known compounds as well as by novel peptides and proteins. Based on its robust performance during the functionality assessment assays, the EndoC-ßH1 cell line was successfully used as a screening platform for identification of novel anti-diabetic drug candidates.

Levels and Patterns of Objectively Measured Physical Activity and Sedentary Time in 4-Year-Old Swedish Children.

BACKGROUND: Levels of physical activity (PA) affect health already at 4 years of age. The aims of this study were to describe levels and patterns of PA and sedentary time (ST) in a sample of 4-year-old Swedish children and to assess the number of children achieving PA guidelines throughout the week. METHODS: Data from 540 4-year-old children enrolled in the population-based PRIMROSE trial was used. PA was measured for a period of 1 week by the Actigraph GT3X+ accelerometer. Average PA, time spent in light PA, moderate-to-vigorous PA (MVPA) and ST were assessed. RESULTS: On average children spent 6.7% of the day in MVPA and 45% of the day being sedentary and 33% (n = 178) of the children met the PA guidelines of 60 minutes of MVPA per day. Boys spent 56.8 (SD 21.8) minutes/day in MVPA, while girls spent 43.0 (SD 18.1) minutes/day in MVPA (P < .001). CONCLUSIONS: Four-year-old children spent almost half of the day being sedentary and only one-third meet the recommended PA guidelines. This finding is alarming as higher levels of PA, already at 4 years of age, seem to reduce the risk of childhood obesity and provides long-term health benefits.

Associations Between Swedish Mothers' and 3- and 5-Year-Old Children's Food Intake.

OBJECTIVE: To investigate associations between mothers' and children's food intake. DESIGN: Cross-sectional study. Background variables collected through self-reports and from the register of the total population. Mothers recorded their own and their children's food intake in a diary during 2 4-day periods. SETTING: Eight counties in mid Sweden. PARTICIPANTS: Three- and 5-year-old children and their mothers were randomly selected from the register of the total population. A total of 2,045 families were invited, 355 of whom accepted. Mothers who accepted were older and to a larger extent born in Sweden. The final sample of mother-child pairs with complete food records was 189. MAIN OUTCOME MEASURES: Mothers' and children's food intake (16 food items). ANALYSIS: Spearman rank-order correlation with 95% confidence intervals (2-sided). Moderation was investigated using generalized estimation equations with robust variance. RESULTS: The strongest correlations between mothers' and children's food intake were found for pizza and oily fish (r = .70-.80). The weakest correlations were found for sugared drinks and fruit and berries (r = .24-.26). Children's age moderated the relationship between mothers' and children's intake of savoury snacks, as did place of residence for pizza intake. CONCLUSIONS AND IMPLICATIONS: There were substantial correlations between children's and mothers' intake of various foods. Modeling of mothers' intake might be more effective in influencing young children's intake of certain foods, whereas other strategies, such as encouraging parents to influence food availability (eg, gatekeeping), might be more useful for some foods.