Characterization of the mutation causative for autosomal recessive hereditary nephropathy in the english cocker spaniel and analysis of gene expression in multiple models of hereditary nephropathy

The domestic dog, Canis familiaris, has over 450 naturally occurring inherited diseases. Over half of these diseases are clinically similar to human diseases making the dog an excellent model in which to study human hereditary diseases. Alport syndrome (AS), a group of heterogeneous, hereditary renal diseases, is one example of such a human disease. The disease is transmitted in three fashions: X-linked, autosomal recessive, and autosomal dominant. AS is caused by mutations in COL4α3, COL4α4 or COL4α5, all members of the type IV collagen family. The proteins products of these genes along with those of the other type IV collagen family members (COL4α1, COL4α2, and COL4α6) are structural components of basement membranes throughout the body. This dissertation describes the measurement of mRNA transcripts in two canine models of AS: a mixed breed model of X-linked AS (XLAS) and the English Cocker Spaniel (ECS) model of autosomal recessive AS (ARAS). The work done revealed a decrease in COL4α4 transcripts. The similarity between the decrease of COL4α5 in the XLAS model and that for COL4α4 in the ARAS model lead to the investigation of COL4α4 as the gene harboring the mutation causative for ARAS in the ECS. Upon sequencing COL4α4, the causative mutation was determined to be an A to T transversion in exon 3. To provide an in vitro model to study type IV collagens, a protocol was designed and experimentally validated to isolate and culture canine Sertoli cells. Canine testes cells were isolated and cultured. Cells were verified as Sertoli cells through positive identification of both SOX9 and Clusterin B proteins, along with sequence verification of SOX9 transcripts. This in vitro model provides a tool to further study the type IV collagens. Overall, the research described herein lead to the identification of the mutation causative for ARAS in the ECS. With this knowledge a genetic test was developed to test for the disease. This research also provided valuable information about the transcript levels of type IV collagens in two models of AS, and provided a novel model in which to study the type IV collagens further

Expanding the Hyper-Enabled Operator Technology across the Special Forces Enterprise

The Hyper-Enabled Operator (HEO) system is the next-generation Special Forces system that will increase the survivability and lethality of operators by providing the right person with the right information at the right time. This system was originally intended for direct-action operators; however, the need for information is common to many Special Forces jobs, including joint terminal air controllers, helicopter pilots, helicopter crew chiefs, intelligence officers, psyops officers, civil affairs officers, and vehicle drivers. This analysis set out to determine the applicability of HEO technology to these eight different positions. First, the HEO system was analyzed to identify the technologies that will play a role in the system. Stakeholder analysis then provided insights into each job, allowing for the determination of their capability gaps. These capability gaps were then aligned against HEO technology. The analysis revealed that several high-level requirements should be added to the HEO system to make it adaptable across the Special Forces enterprise

ATRX is Required for Maintenance of the Neuroprogenitor Cell Pool in the Embryonic Mouse Brain

Mutations in the alpha-thalassemia mental retardation X-linked (ATRX) gene cause a spectrum of abnormalities including intellectual disability, developmental delay, seizures, and microcephaly. The ATRX protein is highly enriched at heterochromatic repetitive sequences adjacent to the centromere, and ATRX depletion results in chromosome congression, segregation, and cohesion defects. Here, we show that Cre-mediated inactivation of Atrx in the embryonic mouse (Mus musculus) brain results in expansion of cerebral cortical layer VI, and a concurrent thinning of layers II-IV. We observed increased cell cycle exit during early-mid neurogenesis, and a depletion of apical progenitors by late neurogenesis in the Atrx-null neocortex, explaining the disproportionate layering. Premature differentiation was associated with an increased generation of outer radial glia (oRG) and TBR2-expressing basal progenitors, as well as increased generation of early-born post-mitotic projection neurons. Atrx deletion also reduced the fidelity of mitotic spindle orientation in apical progenitors, where mutant cells were often oriented at non-parallel angles of division relative to the ventricular surface. We conclude that ATRX is required for correct lamination of the mouse neocortex by regulating the timing of neuroprogenitor cell differentiation

D3.2 Cost Concept Model and Gateway Specification

This document introduces a Framework supporting the implementation of a cost concept model against which current and future cost models for curating digital assets can be benchmarked. The value built into this cost concept model leverages the comprehensive engagement by the 4C project with various user communities and builds upon our understanding of the requirements, drivers, obstacles and objectives that various stakeholder groups have relating to digital curation. Ultimately, this concept model should provide a critical input to the development and refinement of cost models as well as helping to ensure that the curation and preservation solutions and services that will inevitably arise from the commercial sector as ‘supply’ respond to a much better understood ‘demand’ for cost-effective and relevant tools. To meet acknowledged gaps in current provision, a nested model of curation which addresses both costs and benefits is provided. The goal of this task was not to create a single, functionally implementable cost modelling application; but rather to design a model based on common concepts and to develop a generic gateway specification that can be used by future model developers, service and solution providers, and by researchers in follow-up research and development projects.<p></p> The Framework includes:<p></p> • A Cost Concept Model—which defines the core concepts that should be included in curation costs models;<p></p> • An Implementation Guide—for the cost concept model that provides guidance and proposes questions that should be considered when developing new cost models and refining existing cost models;<p></p> • A Gateway Specification Template—which provides standard metadata for each of the core cost concepts and is intended for use by future model developers, model users, and service and solution providers to promote interoperability;<p></p> • A Nested Model for Digital Curation—that visualises the core concepts, demonstrates how they interact and places them into context visually by linking them to A Cost and Benefit Model for Curation.<p></p> This Framework provides guidance for data collection and associated calculations in an operational context but will also provide a critical foundation for more strategic thinking around curation such as the Economic Sustainability Reference Model (ESRM).<p></p> Where appropriate, definitions of terms are provided, recommendations are made, and examples from existing models are used to illustrate the principles of the framework

Validation of Quantitative HPLC Method for Bacosides in KeenMind

Brahmi (Bacopa monnieri) has been used by Ayurvedic medical practitioners in India for almost 3000 years. The pharmacological properties of Bacopa monnieri were studied extensively and the activities were attributed mainly due to the presence of characteristic saponins called “bacosides.” Bacosides are complex mixture of structurally closely related compounds, glycosides of either jujubogenin or pseudojujubogenin. The popularity of herbal medicines and increasing clinical evidence to support associated health claims require standardisation of the phytochemical actives contained in these products. However, unlike allopathic medicines which typically contain a single active compound, herbal medicines are typically complex mixtures of various phytochemicals. The assay for bacosides in the British Pharmacopoeia monograph for Bacopa monnieri exemplifies that only a subset of bacosides present are included in the calculation of total bacosides. These results in calculated bacoside values are significantly lower than those attained for the same material using more inclusive techniques such as UV spectroscopy. This study illustrates some of the problems encountered when applying chemical analysis for standardisation of herbal medicines, particularly in relation to the new method development and validation of bacosides from KeenMind

Unlocking the Keyhole - H2 and PAH emission from molecular clumps in the Keyhole Nebula

To better understand the environment surrounding CO emission clumps in the Keyhole Nebula, we have made images of the region in H2 1-0 S(1) (2.122 um) emission and polycyclic aromatic hydrocarbon (PAH) emission at 3.29 um. Our results show that the H2 and PAH emission regions are morphologically similar, existing as several clumps, all of which correspond to CO emission clumps and dark optical features. The emission confirms the existence of photodissociation regions (PDRs) on the surface of the clumps. By comparing the velocity range of the CO emission with the optical appearance of the H2 and PAH emission, we present a model of the Keyhole Nebula in which the most negative velocity clumps are in front of the ionization region, the clumps at intermediate velocities are in it, and those which have the least negative velocities are at the far side. It may be that these clumps, which appear to have been swept up from molecular gas by the stellar winds from eta Car, are now being over-run by the ionization region and forming PDRs on their surfaces. These clumps comprise the last remnants of the ambient molecular cloud around eta Car.Comment: 8 pages, 4 figures, to be published in MNRA

Implementing Team-Based Nursing to Improve Patient & Staff Satisfaction

https://digitalcommons.psjhealth.org/summit_all/1068/thumbnail.jp

Influence of Crohn’s disease related polymorphisms in innate immune function on ileal microbiome

We have previously identified NOD2 genotype and inflammatory bowel diseases (IBD) phenotype, as associated with shifts in the ileal microbiome (“dysbiosis”) in a patient cohort. Here we report an integrative analysis of an expanded number of Crohn's disease (CD) related genetic defects in innate immune function (NOD2, ATG16L1, IRGM, CARD9, XBP1, ORMDL3) and composition of the ileal microbiome by combining the initial patient cohort (Batch 1, 2005–2010, n = 165) with a second consecutive patient cohort (Batch 2, 2010–2012, n = 118). These combined patient cohorts were composed of three non-overlapping phenotypes: 1.) 106 ileal CD subjects undergoing initial ileocolic resection for diseased ileum, 2.) 88 IBD colitis subjects without ileal disease (predominantly ulcerative colitis but also Crohn’s colitis and indeterminate colitis, and 3.) 89 non-IBD subjects. Significant differences (FDR C. difficile infection, and NOD2 genotype on ileal dysbiosis in the expanded analysis. The relative abundance of the Proteobacteria phylum was positively associated with ileal CD and colitis phenotypes, but negatively associated with NOD2R genotype. Additional associations with ORMDL3 and XBP1 were detected at the phylum/subphylum level. IBD medications, such as immunomodulators and anti-TNFα agents, may have a beneficial effect on reversing dysbiosis associated with the IBD phenotype. Exploratory analysis comparing microbial composition of the disease unaffected region of the resected ileum between 27 ileal CD patients who subsequently developed endoscopic recurrence within 6–12 months versus 34 patients who did not, suggested that microbial biomarkers in the resected specimen helped stratify patients with respect to risk of post-surgical recurrence.</div

A Mg(OH)2 coprecipitation method for determining chromium speciation and isotopic composition in seawater

Chromium (Cr) stable isotopes have emerged as a powerful tool for tracking environmental redox transfor- mations. This is because Cr isotopes are fractionated during redox reactions between Cr(III) and Cr(VI). In order to fully exploit the information recorded within Cr isotope compositions, we must be able to track changes in Cr speciation throughout the environment and, in particular, the changes in speciation between input to the ocean and eventual deposition in sediments. We must also be able to access the isotope compositions of each Cr species, rather than only total dissolved Cr. We have thus developed a magnesium hydroxide coprecipitation method that meets these objectives. This method achieves complete recovery and has a typical precision on concentration measurements of !8% (1σ). It was tested using seawater collected from Saanich Inlet, a persis- tently anoxic fjord on the Pacific coast of Canada. Chromium speciation profiles and proof-of-concept isotope ratio measurements on selected samples indicate that isotopically lighter Cr(III) can be isolated from coexisting isotopically heavier Cr(VI), effectively resolving species-specific Cr isotope compositions. While the oxygenated surface waters of Saanich Inlet follow the generally observed correlation between seawater Cr concentration and its isotopic composition, seawater from anoxic depths diverges from this array, indicating that different pro- cesses are responsible for setting the isotope composition of these deeper waters. Broader application of Mg(OH)2 coprecipitation has strong potential to yield new insights into the fractionation of Cr isotopes in the oceans and the pathways that ultimately set the Cr isotopic composition of marine sediments and sedimentary archives