Identification of H19 polymorphism for an assessment of biallelic expression

Abstract only availableAnimals produced from assisted reproductive technologies suffer from developmental abnormalities and early fetal death at a higher frequency than that observed in those produced by natural breeding. These symptoms are reminiscent of imprinting disruptions, suggesting the possibility of an alteration in the expression of imprinted genes such as biallelic expression or silencing. H19 is one of the imprinted genes first identified in mice and humans, but its imprinting status has not been determined in pigs. The objective of this study was to identify an H19 polymorphism and estimate its frequency in the commercial pig population. In this study a polymorphism in the H19 gene was identified. The PCR products contained a pooled genome with over 900 specimens to support this finding. From the positive PCR products, the DNA was cloned and transformed with a TOPO TA Cloning kit (Invitrogen). Positive colonies were identified and digested with an AciI enzyme, which cut the DNA in specific fragments that were identifiable in a gel. Analysis of the gel showed evidence that a polymorphism exists on the H19 gene.F.B. Miller Undergraduate Research Program in Animal Science

Swine models of Human Conditions

Comparative Medicine: Innovations for the Animal to Human Health CorridorSince the introduction of Dolly, the first cloned sheep, genetic engineering of livestock has advanced to the point that nearly any genetic modification appears feasible. Genes can be introduced, deleted or altered in cultured cells. These cells can be evaluated in culture and then used to regenerate live animals by somatic cell nuclear transfer (cloning). Since pigs share similar physiology with humans and have proven to be excellent models for many human conditions, genetic engineering and animal cloning are being used to produce models of human disease states and disorders. These models are proving to be particularly important for situations where laboratory rodents have proven to be inadequate. Current successes of genetically engineered swine biomedical models will be presented

Short-Haul Revitalization Study Final Report

A feasibility study was performed for an advanced commercial short-haul aircraft to evaluate the potential for increased service for short-haul flights that operate out of regional and community airports. An analysis of potential origin-destination markets and trip distances resulted in a seat capacity selection of 48 passengers and a design range of 600 NM. A down-select of advanced technologies resulted in a hybrid-electric propulsion system being chosen as the primary enabling technology. A conceptual design of the advanced aircraft was developed, and a mission and sizing analysis was performed, comparing variants of the advanced aircraft with different levels of electrification. Fairly aggressive levels of electrification and battery specific energy are needed for the hybridelectric architecture to realize any benefit in terms of total energy cost for the 600 NM design mission. The development and operational costs were estimated for the advanced aircraft and compared to the baseline. This analysis demonstrated the negative effect of the cost to develop the hybrid-electric technology on the eventual operating cost. A market analysis was performed to determine possible passenger demand for the advanced shorthaul aircraft. According to the market analysis, there is potential demand for such an aircraft, but not necessarily in many of the smaller regional and community airports that were the intended beneficiaries of this new aircraft concept

Clinically Actionable Hypercholesterolemia and Hypertriglyceridemia in Children with Nonalcoholic Fatty Liver Disease

OBJECTIVE: To determine the percentage of children with nonalcoholic fatty liver disease (NAFLD) in whom intervention for low-density lipoprotein cholesterol or triglycerides was indicated based on National Heart, Lung, and Blood Institute guidelines. STUDY DESIGN: This multicenter, longitudinal cohort study included children with NAFLD enrolled in the National Institute of Diabetes and Digestive and Kidney Diseases Nonalcoholic Steatohepatitis Clinical Research Network. Fasting lipid profiles were obtained at diagnosis. Standardized dietary recommendations were provided. After 1 year, lipid profiles were repeated and interpreted according to National Heart, Lung, and Blood Institute Expert Panel on Integrated Guidelines for Cardiovascular Health and Risk Reduction. Main outcomes were meeting criteria for clinically actionable dyslipidemia at baseline, and either achieving lipid goal at follow-up or meeting criteria for ongoing intervention. RESULTS: There were 585 participants, with a mean age of 12.8 years. The prevalence of children warranting intervention for low-density lipoprotein cholesterol at baseline was 14%. After 1 year of recommended dietary changes, 51% achieved goal low-density lipoprotein cholesterol, 27% qualified for enhanced dietary and lifestyle modifications, and 22% met criteria for pharmacologic intervention. Elevated triglycerides were more prevalent, with 51% meeting criteria for intervention. At 1 year, 25% achieved goal triglycerides with diet and lifestyle changes, 38% met criteria for advanced dietary modifications, and 37% qualified for antihyperlipidemic medications. CONCLUSIONS: More than one-half of children with NAFLD met intervention thresholds for dyslipidemia. Based on the burden of clinically relevant dyslipidemia, lipid screening in children with NAFLD is warranted. Clinicians caring for children with NAFLD should be familiar with lipid management

The Long-Baseline Neutrino Experiment: Exploring Fundamental Symmetries of the Universe

The preponderance of matter over antimatter in the early Universe, the dynamics of the supernova bursts that produced the heavy elements necessary for life and whether protons eventually decay --- these mysteries at the forefront of particle physics and astrophysics are key to understanding the early evolution of our Universe, its current state and its eventual fate. The Long-Baseline Neutrino Experiment (LBNE) represents an extensively developed plan for a world-class experiment dedicated to addressing these questions. LBNE is conceived around three central components: (1) a new, high-intensity neutrino source generated from a megawatt-class proton accelerator at Fermi National Accelerator Laboratory, (2) a near neutrino detector just downstream of the source, and (3) a massive liquid argon time-projection chamber deployed as a far detector deep underground at the Sanford Underground Research Facility. This facility, located at the site of the former Homestake Mine in Lead, South Dakota, is approximately 1,300 km from the neutrino source at Fermilab -- a distance (baseline) that delivers optimal sensitivity to neutrino charge-parity symmetry violation and mass ordering effects. This ambitious yet cost-effective design incorporates scalability and flexibility and can accommodate a variety of upgrades and contributions. With its exceptional combination of experimental configuration, technical capabilities, and potential for transformative discoveries, LBNE promises to be a vital facility for the field of particle physics worldwide, providing physicists from around the globe with opportunities to collaborate in a twenty to thirty year program of exciting science. In this document we provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess.Comment: Major update of previous version. This is the reference document for LBNE science program and current status. Chapters 1, 3, and 9 provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess. 288 pages, 116 figure

Genome-wide imputation study identifies novel HLA locus for pulmonary fibrosis and potential role for auto-immunity in fibrotic idiopathic interstitial pneumonia.

To access publisher's full text version of this article, please click on the hyperlink in Additional Links field or click on the hyperlink at the top of the page marked Files. This article is open access.Fibrotic idiopathic interstitial pneumonias (fIIP) are a group of fatal lung diseases with largely unknown etiology and without definitive treatment other than lung transplant to prolong life. There is strong evidence for the importance of both rare and common genetic risk alleles in familial and sporadic disease. We have previously used genome-wide single nucleotide polymorphism data to identify 10 risk loci for fIIP. Here we extend that work to imputed genome-wide genotypes and conduct new RNA sequencing studies of lung tissue to identify and characterize new fIIP risk loci.We performed genome-wide genotype imputation association analyses in 1616 non-Hispanic white (NHW) cases and 4683 NHW controls followed by validation and replication (878 cases, 2017 controls) genotyping and targeted gene expression in lung tissue. Following meta-analysis of the discovery and replication populations, we identified a novel fIIP locus in the HLA region of chromosome 6 (rs7887 P meta  = 3.7 × 10(-09)). Imputation of classic HLA alleles identified two in high linkage disequilibrium that are associated with fIIP (DRB1*15:01 P = 1.3 × 10(-7) and DQB1*06:02 P = 6.1 × 10(-8)). Targeted RNA-sequencing of the HLA locus identified 21 genes differentially expressed between fibrotic and control lung tissue (Q < 0.001), many of which are involved in immune and inflammatory response regulation. In addition, the putative risk alleles, DRB1*15:01 and DQB1*06:02, are associated with expression of the DQB1 gene among fIIP cases (Q < 1 × 10(-16)).We have identified a genome-wide significant association between the HLA region and fIIP. Two HLA alleles are associated with fIIP and affect expression of HLA genes in lung tissue, indicating that the potential genetic risk due to HLA alleles may involve gene regulation in addition to altered protein structure. These studies reveal the importance of the HLA region for risk of fIIP and a basis for the potential etiologic role of auto-immunity in fIIP.National Heart, Lung and Blood Institute R01-HL095393 R01-HL097163 P01-HL092870 RC2-HL101715 U01-HL089897 U01-HL089856 U01-HL108642 P50-HL089493

Combination therapy in a xenograft model of glioblastoma: enhancement of the antitumor activity of temozolomide by an MDM2 antagonist

OBJECTIVE Improvement in treatment outcome for patients with glioblastoma multiforme (GBM) requires a multifaceted approach due to dysregulation of numerous signaling pathways. The murine double minute 2 (MDM2) protein may fulfill this requirement because it is involved in the regulation of growth, survival, and invasion. The objective of this study was to investigate the impact of modulating MDM2 function in combination with front-line temozolomide (TMZ) therapy in GBM. METHODS The combination of TMZ with the MDM2 protein-protein interaction inhibitor nutlin3a was evaluated for effects on cell growth, p53 pathway activation, expression of DNA repair proteins, and invasive properties. In vivo efficacy was assessed in xenograft models of human GBM. RESULTS In combination, TMZ/nutlin3a was additive to synergistic in decreasing growth of wild-type p53 GBM cells. Pharmacodynamic studies demonstrated that inhibition of cell growth following exposure to TMZ/nutlin3a correlated with: 1) activation of the p53 pathway, 2) downregulation of DNA repair proteins, 3) persistence of DNA damage, and 4) decreased invasion. Pharmacokinetic studies indicated that nutlin3a was detected in human intracranial tumor xenografts. To assess therapeutic potential, efficacy studies were conducted in a xenograft model of intracranial GBM by using GBM cells derived from a recurrent wild-type p53 GBM that is highly TMZ resistant (GBM10). Three 5-day cycles of TMZ/nutlin3a resulted in a significant increase in the survival of mice with GBM10 intracranial tumors compared with single-agent therapy. CONCLUSIONS Modulation of MDM2/p53-associated signaling pathways is a novel approach for decreasing TMZ resistance in GBM. To the authors' knowledge, this is the first study in a humanized intracranial patient-derived xenograft model to demonstrate the efficacy of combining front-line TMZ therapy and an inhibitor of MDM2 protein-protein interactions

Oceanographic Structure and Light Levels Drive Patterns of Sound Scattering Layers in a Low-Latitude Oceanic System

Several factors have been reported to structure the spatial and temporal patterns of sound scattering layers, including temperature, oxygen, salinity, light, and physical oceanographic conditions. In this study, we examined the spatiotemporal variability of acoustically detected sound scattering layers in the northern Gulf of Mexico to investigate the drivers of this variability, including mesoscale oceanographic features [e.g., Loop Current-origin water (LCOW), frontal boundaries, and Gulf Common Water]. Results indicate correlations in the vertical position and acoustic backscatter intensity of sound scattering layers with oceanographic conditions and light intensity. LCOW regions displayed consistent decreases, by a factor of two and four, in acoustic backscatter intensity in the upper 200 m relative to frontal boundaries and Gulf Common Water, respectively. Sound scattering layers had greater backscatter intensity at night in comparison to daytime (25x for frontal boundaries, 17x for LCOW, and 12x for Gulf Common Water). The importance of biotic (primary productivity) and abiotic (sea surface temperature, salinity) factors varied across oceanographic conditions and depth intervals, suggesting that the patterns in distribution and behavior of mesopelagic assemblages in low-latitude, oligotrophic ecosystems can be highly dynamic

Developing the principles of chair based exercise for older people: a modified Delphi study

Background Chair based exercise (CBE) is suggested to engage older people with compromised health and mobility in an accessible form of exercise. A systematic review looking at the benefits of CBE for older people identified a lack of clarity regarding a definition, delivery, purpose and benefits. This study aimed to utilise expert consensus to define CBE for older people and develop a core set of principles to guide practice and future research. Methods The framework for consensus was constructed through a team workshop identifying 42 statements within 7 domains. A four round electronic Delphi study with multi-disciplinary health care experts was undertaken. Statements were rated using a 5 point Likert scale of agreement and free text responses. A threshold of 70% agreement was used to determine consensus. Free text responses were analysed thematically. Between rounds a number of strategies (e.g., amended wording of statements, generation and removal of statements) were used to move towards consensus. Results 16 experts agreed on 46 statements over four rounds of consultation (Round 1: 22 accepted, 3 removed, 5 new and 17 modified; Round 2: 16 accepted, 0 removed, 4 new and 6 modified; Round 3: 4 accepted, 2 removed, 0 new and 4 modified; Round 4: 4 accepted, 0 removed, 0 new, 0 modified). Statements were accepted in all seven domains: the definition of CBE (5), intended users (3), potential benefits (8), structure (12), format (8), risk management (7) and evaluation (3). The agreed definition of CBE had five components: 1. CBE is primarily a seated exercise programme; 2. The purpose of using a chair is to promote stability in both sitting and standing; 3. CBE should be considered as part of a continuum of exercise for frail older people where progression is encouraged; 4. CBE should be used flexibly to respond to the changing needs of frail older people; and 5. Where possible CBE should be used as a starting point to progress to standing programmes. Conclusions Consensus has been reached on a definition and a set of principles governing CBE for older people; this provides clarity for implementation and future research about CBE

Abundance and Occurrence of Common Bottlenose Dolphins (Tursiops truncatus) in Three Estuaries of the Northwestern Gulf of Mexico

Current abundance estimates for populations of common bottlenose dolphins (Tursiops truncatus, Montagu, 1821) in bays, sounds, and estuaries are lacking throughout most of the northwestern Gulf of Mexico, including areas of Texas and western Louisiana. To address this issue, we conducted 92 small-boat photographic identification surveys covering ~2000 km2 and comprising ~11,000 km of track-line in winter and summer seasons in West Bay, TX (2014 – 2015, n = 25), the Galveston Bay, TX system (2016, n = 50), Sabine Lake, TX (2017, n = 17), and adjacent coastal waters. Individual dolphin encounter histories were constrained by spatiotemporal parameters to approximately represent 1) a “Bay” estimate of individuals limited to the interior of each embayment, and 2) a “Selective” estimate of the number of individuals in each survey area (including nearshore coastal waters), filtered for potential transient dolphins. Using the Selective dataset, estimated dolphins (95% CI) were (winter and summer, respectively) 38 (29 – 47) and 37 (33 – 40) for West Bay, 842 (694 – 990) and 1132 (846 – 1417) for Galveston Bay, and 122 (73 – 170) and 162 (114 – 210) for Sabine Lake. A range of 4-15% of marked individuals in each study area were identified as inter-bay matches. These results provide new insights on the potential spatial range of each population, update previous abundance estimates for West Bay and Galveston Bay, and contribute novel population information for Sabine Lake and adjacent coastal waters of the northwestern Gulf of Mexico