Identification of the Functional Variant(s) that Explain the Low-Density Lipoprotein Receptor (LDLR) GWAS SNP rs6511720 Association with Lower LDL-C and Risk of CHD

BACKGROUND: The Low-Density Lipoprotein Receptor (LDLR) SNP rs6511720 (G>T), located in intron-1 of the gene, has been identified in genome-wide association studies (GWAS) as being associated with lower plasma levels of LDL-C and a lower risk of coronary heart disease (CHD). Whether or not rs6511720 is itself functional or a marker for a functional variant elsewhere in the gene is not known. METHODS: The association of LDLR SNP rs6511720 with incidence of CHD and levels of LDL-C was determined by reference to CARDIoGRAM, C4D and Global lipids genetics consortium (GLGC) data. SNP annotation databases were used to identify possible SNP function and prioritization. Luciferase reporter assays in the liver cell line Huh7 were used to measure the effect of variant genotype on gene expression. Electrophoretic Mobility Shift Assays (EMSAs) were used to identify the Transcription Factors (TFs) involved in gene expression regulation. RESULTS: The phenotype-genotype analysis showed that the rs6511720 minor allele is associated with lower level of LDL-C [beta = -0.2209, p = 3.85 x10-262], and lower risk of CHD [log (OR) = 0.1155, p = 1.04 x10-7]. Rs6511720 is in complete linkage. Rs6511720 is in complete linkage disequilibrium (LD) with three intron-1 SNPs (rs141787760, rs60173709, rs57217136). Luciferase reporter assays in Huh7 cells showed that the rare alleles of both rs6511720 and rs57217136 caused a significant increase in LDLR expression compared to the common alleles (+29% and +24%, respectively). Multiplex Competitor-EMSAs (MC-EMSA) identified that the transcription factor serum response element (SRE) binds to rs6511720, while retinoic acid receptor (RAR) and signal transducer and activator of transcription 1 (STAT1) bind to rs57217136. CONCLUSION: Both LDLR rs6511720 and rs57217136 are functional variants. Both these minor alleles create enhancer-binding protein sites for TFs and may contribute to increased LDLR expression, which is consequently associated with reduced LDL-C levels and 12% lower CHD risk

Demonstration of the Presence of the "Deleted" MIR122 Gene in HepG2 Cells

MicroRNA 122 (miR-122) is highly expressed in the liver where it influences diverse biological processes and pathways, including hepatitis C virus replication and metabolism of iron and cholesterol. It is processed from a long non-coding primary transcript (~7.5 kb) and the gene has two evolutionarily-conserved regions containing the pri-mir-122 promoter and pre-mir-122 hairpin region. Several groups reported that the widely-used hepatocytic cell line HepG2 had deficient expression of miR-122, previously ascribed to deletion of the pre-mir-122 stem-loop region. We aimed to characterise this deletion by direct sequencing of 6078 bp containing the pri-mir-122 promoter and pre-mir-122 stem-loop region in HepG2 and Huh-7, a control hepatocytic cell line reported to express miR-122, supported by sequence analysis of cloned genomic DNA. In contrast to previous findings, the entire sequence was present in both cell lines. Ten SNPs were heterozygous in HepG2 indicating that DNA was present in two copies. Three validation isolates of HepG2 were sequenced, showing identical genotype to the original in two, whereas the third was different. Investigation of promoter chromatin status by FAIRE showed that Huh-7 cells had 6.2 ± 0.19- and 2.7 ± 0.01- fold more accessible chromatin at the proximal (HNF4α-binding) and distal DR1 transcription factor sites, compared to HepG2 cells (p=0.03 and 0.001, respectively). This was substantiated by ENCODE genome annotations, which showed a DNAse I hypersensitive site in the pri-mir-122 promoter in Huh-7 that was absent in HepG2 cells. While the origin of the reported deletion is unclear, cell lines should be obtained from a reputable source and used at low passage number to avoid discrepant results. Deficiency of miR-122 expression in HepG2 cells may be related to a relative deficiency of accessible promoter chromatin in HepG2 versus Huh-7 cells

Metodologia per a la generació de consens social en el desplegament d'infraestructures de telefonia mòbil

Aquest document de treball es centra específicament en el desenvolupament de la infraestructura de telefonia mòbil a Catalunya com a exemple de tecnologia amb profundes implicacions socials. Esdevé un intent inicial de definir les principals línies de recerca en aquest tema. El projecte que el sosté és emmarcat en el programa de recerca Negrisc, centrat a examinar la negociació del risc en les tecnologies de la informació i de la comunicació. Parteix d'una aproximació que reconeix la natura multidisciplinària del risc i que, per tant, exigeix una aproximació interdisciplinària al seu estudi i a la gestió subsegüent. Aquesta perspectiva emfatitza la necessitat d'integrar els punts de vista dels diferents stakeholders, incloent-hi el públic, en la gestió d'una tecnologia socialment controvertida. Aquest és un pas necessari per a permetre la generació d'algun tipus de consens en el cas del desenvolupament de la infraestructura de telefonia mòbil a Catalunya.Este documento de trabajo se encuentra específicamente centrado en el desarrollo de la infraestructura de telefonía móvil en Cataluña como ejemplo de tecnología con profundas implicaciones sociales. Es un intento inicial de definir las principales líneas de investigación en este tema. El proyecto que lo sostiene se encuentra enmarcado en el programa de investigación Negrisc, centrado en examinar la negociación del riesgo en las tecnologías de la información y de la comunicación. Parte de una aproximación que reconoce la naturaleza multidisciplinaria del riesgo y que, por lo tanto, exige una aproximación interdisciplinaria a su estudio y a la subsiguiente gestión. Esta perspectiva enfatiza la necesidad de integrar los puntos de vista de los stakeholders, incluyendo al público, en la gestión de una tecnología socialmente controvertida. Éste es un paso necesario para permitir la generación de algún tipo de consenso en el caso del desarrollo de la infraestructura de telefonía móvil en Cataluña.This working paper is specifically concerned with the development of the mobile telephone infrastructure in Catalonia as an example of a technology with profound social implications. It is an initial attempt to define certain lines of research related to the topic. This project is operating within the broader Negrisc research programme, which examines the negotiation of risk in the field of information and communication technologies. It sets out from an approach which recognises the multidisciplinary nature of risk and which therefore demands an interdisciplinary approach to its study and subsequent management. This perspective stresses the necessity of integrating viewpoints of the different stakeholders, including the public, in the management of a socially controversial technology. This is seen as a necessary step to enable the generation of some sort of social consensus in the case of the rollout of the mobile phone infrastructure in Catalonia

Leukocyte migration in experimental inflammatory bowel disease

Emigration of leukocytes from the circulation into tissue by transendothelial migration, is mediated subsequently by adhesion molecules such as selectins, chemokines and integrins. This multistep paradigm, with multiple molecular choices at each step, provides a diversity in signals. The influx of neutrophils, monocytes and lymphocytes into inflamed tissue is important in the pathogenesis of chronic inflammatory bowel disease. The importance of each of these groups of adhesion molecules in chronic inflammatory bowel disease, either in human disease or in animal models, will be discussed below. Furthermore, the possibilities of blocking these different steps in the process of leukocyte extravasation in an attempt to prevent further tissue damage, will be taken into account

Stronger than your voices:A cognitive behavioral therapy for youth suffering from auditory verbal hallucinations

Objective: Auditory verbal hallucinations (AVHs) are a common feature in youth and mostly transient. Nevertheless, while present, AVH can cause considerable distress. Children and adolescents seeking help for distressing AVH represent a heterogeneous group in terms of underlying factors, yet they consistently suffer from their AVH. Until now, a youth-specific psychotherapeutic intervention for AVH was lacking. Experts in the field of treating AVH in both adults and youngsters collaborated with service users to develop the cognitive behavioral therapy (CBT) "Stronger Than Your Voices" (STYV). We investigated feasibility and clinical outcomes of the STYV therapy. Methods: Patients were derived from children and adolescents seeking help for AVH at the UMC Utrecht outpatient clinic with an indication for STYV therapy. Therapists preferably originated from referring health care facilities and were required to have sufficient general knowledge and experience with CBT. They received a short individual training to apply STYV. After, patients and their therapists could participate this naturalistic pilot study, assessing feasibility, tolerability, and clinical change when applying the STYV therapy. Results: Six participants (10-16 years old), all suffering from comorbid psychopathology, provided pre and post measures, all completing STYV therapy without experiencing an aggravation of symptoms. AVH total impact decreased 40% with Cohen's d within-group effect size (1.28) also suggesting clinically meaningful change. Therapists were positive about STYV therapy and manual. Conclusion: The STYV therapy is feasible for youth with distressing AVH. First results indicate that STYV may be clinically effective. A trial to further test effectiveness in a larger sample is needed

Visitation Enablers and Barriers: Evaluating the Influences of Practical, Relational, and Experiential Factors on Visitation in Dutch Prisons

This paper aims to advance theory and knowledge about prison visitation by organizing prior studies within a framework of visitation enablers and barriers and examining how practical, relational, and experiential factors explain variation in prison visiting among 773 adult males across eight Dutch prisons. Findings suggest that all three domains play out at once to influence visitation. Whether visitors come to visit seems to depend on their relationship with the incarcerated individual, whereas traveling distance is more predictive of how often they visit. Policies that introduce practical barriers can differentially affect visits from specific relationships. Finally, results indicate that incarcerated individuals make decisions about visits based on their in-prison experiences. Policy and research implications are discussed.Criminal Justice: Legitimacy, accountability, and effectivit

Jak-STAT regulation of cyst stem cell development in the Drosophila testis

Establishment and maintenance of functional stem cells is critical for organ development and tissue homeostasis. Little is known about the mechanisms underlying stem establishment during organogenesis. Drosophila testes are among the most thoroughly characterized systems for studying stem cell behavior, with germline stem cells (GSCs) and somatic cyst stem cells (CySCs) cohabiting a discrete stem cell niche at the testis apex. GSCs and CySCs are arrayed around hub cells that also comprise the niche and communication between hub cells, GSCs, and CySCs regulates the balance between stem cell maintenance and differentiation. Recent data has shown that functional, asymmetrically dividing GSCs are first established at similar to 23 h after egg laying during Drosophila testis morphogenesis (Sheng et al., 2009). This process correlates with coalescence of the hub, but development of CySCs from somatic gonadal precursors (SGPs) was not examined. Here, we show that functional CySCs are present at the time of GSC establishment, and that Jak-STAT signaling is necessary and sufficient for CySC maintenance shortly thereafter. Furthermore, hyper-activation of Jak in CySCs promotes expansion of the GSC population, while ectopic Jak activation in the germline induces GSC gene expression in GSC daughter cells but does not prevent spermatogenic differentiation. Together, these observations indicate that, similar to adult testes, Jak-STAT signaling from the hub acts on both GSCs and CySC to regulate their development and differentiation, and that additional signaling from CySCs to the GSCs play a dominant role in controlling GSC maintenance during niche formation. (c) 2012 Elsevier Inc. All rights reserved

L-Cysteine Containing Vitamin Supplement Which Prevents or Alleviates Alcohol-related Hangover Symptoms : Nausea, Headache, Stress and Anxiety

Correction ALCOHOL AND ALCOHOLISM Volume: 55 Issue: 6 Pages: 705-705 DOI: 10.1093/alcalc/agaa088 Published: NOV 2020Aims: Alcohol-related hangover symptoms: nausea, headache, stress and anxiety cause globally considerable amount of health problems and economic losses. Many of these harmful effects are produced by alcohol and its metabolite, acetaldehyde, which also is a common ingredient in alcohol beverages. The aim of the present study is to investigate the effect of the amino acid L-cysteine on the alcohol/acetaldehyde related aftereffects. Methods: Voluntary healthy participants were recruited through advertisements. Volunteers had to have experience of hangover and/or headache. The hangover study was randomized, double-blind and placebo-controlled. Nineteen males randomly swallowed placebo and L-cysteine tablets. The alcohol dose was 1.5 g/kg, which was consumed during 3 h. Results: The primary results based on correlational analysis showed that L-cysteine prevents or alleviates hangover, nausea, headache, stress and anxiety. For hangover, nausea and headache the results were apparent with the L-cysteine dose of 1200 mg and for stress and anxiety already with the dose of 600 mg. Conclusions: L-cysteine would reduce the need of drinking the next day with no or less hangover symptoms: nausea, headache, stress and anxiety. Altogether, these effects of L-cysteine are unique and seem to have a future in preventing or alleviating these harmful symptoms as well as reducing the risk of alcohol addiction.Peer reviewe

Superimposed tissue formation in human aortic valve disease: differences between regurgitant and stenotic valves

The formation of superimposed tissue (SIT), a layer on top of the original valve leaflet, has been described in patients with mitral regurgitation as a major contributor to valve thickening and possibly as a result of increased mechanical stresses. However, little is known whether SIT formation also occurs in aortic valve disease. We therefore performed histological analyses to assess SIT formation in aortic valve leaflets (n = 31) from patients with aortic stenosis (n = 17) or aortic regurgitation due to aortic dilatation (n = 14). SIT was observed in both stenotic and regurgitant aortic valves, both on the ventricular and aortic sides, but with significant differences in distribution and composition. Regurgitant aortic valves showed more SIT formation in the free edge, leading to a thicker leaflet at that level, while stenotic aortic valves showed relatively more SIT formation on the aortic side of the body part of the leaflet. SIT appeared to be a highly active area, as determined by large populations of myofibroblasts, with varied extracellular matrix composition (higher collagen content in stenotic valves). Further, the identification of the SIT revealed the presence of foldings of the free edge in the diseased aortic valves. Insights into SIT regulation may further help in understanding the pathophysiology of aortic valve disease and potentially lead to the development of new therapeutic treatments.Cardiolog