Validation of an enzyme- linked immunoassay assay for osteocalcin, a marker of bone formation, in dried blood spots

ObjectivesInvestigating factors that contribute to bone loss and accretion across populations in remote settings is challenging, particularly where diagnostic tools are scarce. To mitigate this challenge, we describe validation of a commercial ELISA assay to measure osteocalcin, a biomarker of bone formation, from dried blood spots (DBS).MethodsWe validated the Osteocalcin Human SimpleStep ELISA kit from Abcam (ab1951214) using 158 matched plasma and DBS samples. Passing- Bablok regression analysis assessed the relationships between plasma and DBS osteocalcin concentrations. Dilutional linearity and spike and recovery experiments determined if the DBS matrix interfered with osteocalcin measurement, and intra- and inter- assay coefficients of variation (CVs) were calculated. Limit of detection, analyte stability, and specific forms of osteocalcin measured by the kit were also investigated.ResultsMean plasma osteocalcin value was 218.2 ng/mL (range 64.6- 618.1 ng/mL). Linear relationships existed between plasma and DBS concentrations of osteocalcin, with no apparent bias in plasma vs DBS concentrations. There was no apparent interference of the DBS matrix with measurement of osteocalcin in DBS. Intra- assay CV for DBS was ~8%, while average inter- assay CV was 14.8%. Limit of detection was 0.34- ng/mL. Osteocalcin concentrations were stable in DBS stored at - 28°C and room temperature, but not those stored at 37°C. This ELISA kit detects total osteocalcin.ConclusionsOsteocalcin, a bone formation biomarker, can be measured from DBS. Combined with a previously validated DBS assay for TRACP- 5b, a bone resorption biomarker, these assays have the potential to help researchers disentangle the many factors contributing to bone strength.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/162811/2/ajhb23394.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/162811/1/ajhb23394_am.pd

Transition from initiation to promoter proximal pausing requires the CTD of RNA polymerase II

The C-terminal domain (CTD) of mammalian RNA polymerase II consists of 52 repeats of the consensus hepta-peptide YSPTSPS, and links transcription to the processing of pre-mRNA. Although Pol II with a CTD shortened to five repeats (Pol II Δ5) is transcriptionally inactive on chromatin templates, it is not clear whether CTD is required for promoter recognition in vivo. Here, we demonstrate that in the context of chromatin, Pol II Δ5 can bind to the c-myc promoter with the same efficiency as wild type Pol II. However, Pol II Δ5 does not form a stable initiation complex, and does not transcribe promoter proximal sequences. Fluorescence recovery after photobleaching (FRAP) experiments with cells expressing enhanced green fluorescent protein (EGFP)-tagged Δ5 or wildtype Pol II revealed a single, highly mobile Pol II Δ5 fraction whereas wildtype Pol II yielded less mobile fractions. These data suggest that CTD is not required for promoter recognition, but rather for subsequent formation of a stable initiation complex and isomerization to an elongation competent complex

Finite flavour groups of fermions

We present an overview of the theory of finite groups, with regard to their application as flavour symmetries in particle physics. In a general part, we discuss useful theorems concerning group structure, conjugacy classes, representations and character tables. In a specialized part, we attempt to give a fairly comprehensive review of finite subgroups of SO(3) and SU(3), in which we apply and illustrate the general theory. Moreover, we also provide a concise description of the symmetric and alternating groups and comment on the relationship between finite subgroups of U(3) and finite subgroups of SU(3). Though in this review we give a detailed description of a wide range of finite groups, the main focus is on the methods which allow the exploration of their different aspects.Comment: 89 pages, 6 figures, some references added, rearrangement of part of the material, section on SU(3) subgroups substantially extended, some minor revisions. Version for publication in J. Phys. A. Table 12 corrected to match eq.(256), table 14 and eq.(314) corrected to match the 2-dimensional irreps defined on p.6

The Evolution of Bat Vestibular Systems in the Face of Potential Antagonistic Selection Pressures for Flight and Echolocation

PMCID: PMC3634842This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

The nuclear receptors of Biomphalaria glabrata and Lottia gigantea: Implications for developing new model organisms

© 2015 Kaur et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedNuclear receptors (NRs) are transcription regulators involved in an array of diverse physiological functions including key roles in endocrine and metabolic function. The aim of this study was to identify nuclear receptors in the fully sequenced genome of the gastropod snail, Biomphalaria glabrata, intermediate host for Schistosoma mansoni and compare these to known vertebrate NRs, with a view to assessing the snail's potential as a invertebrate model organism for endocrine function, both as a prospective new test organism and to elucidate the fundamental genetic and mechanistic causes of disease. For comparative purposes, the genome of a second gastropod, the owl limpet, Lottia gigantea was also investigated for nuclear receptors. Thirty-nine and thirty-three putative NRs were identified from the B. glabrata and L. gigantea genomes respectively, based on the presence of a conserved DNA-binding domain and/or ligand-binding domain. Nuclear receptor transcript expression was confirmed and sequences were subjected to a comparative phylogenetic analysis, which demonstrated that these molluscs have representatives of all the major NR subfamilies (1-6). Many of the identified NRs are conserved between vertebrates and invertebrates, however differences exist, most notably, the absence of receptors of Group 3C, which includes some of the vertebrate endocrine hormone targets. The mollusc genomes also contain NR homologues that are present in insects and nematodes but not in vertebrates, such as Group 1J (HR48/DAF12/HR96). The identification of many shared receptors between humans and molluscs indicates the potential for molluscs as model organisms; however the absence of several steroid hormone receptors indicates snail endocrine systems are fundamentally different.The National Centre for the Replacement, Refinement and Reduction of Animals in Research, Grant Ref:G0900802 to CSJ, LRN, SJ & EJR [www.nc3rs.org.uk]

Three-Dimensional Analysis of Spiny Dendrites Using Straightening and Unrolling Transforms

Current understanding of the synaptic organization of the brain depends to a large extent on knowledge about the synaptic inputs to the neurons. Indeed, the dendritic surfaces of pyramidal cells (the most common neuron in the cerebral cortex) are covered by thin protrusions named dendritic spines. These represent the targets of most excitatory synapses in the cerebral cortex and therefore, dendritic spines prove critical in learning, memory and cognition. This paper presents a new method that facilitates the analysis of the 3D structure of spine insertions in dendrites, providing insight on spine distribution patterns. This method is based both on the implementation of straightening and unrolling transformations to move the analysis process to a planar, unfolded arrangement, and on the design of DISPINE, an interactive environment that supports the visual analysis of 3D patterns

Design of a graphical and interactive interface for facilitating access to drug contraindications, cautions for use, interactions and adverse effects

<p>Abstract</p> <p>Background</p> <p>Drug iatrogeny is important but could be decreased if contraindications, cautions for use, drug interactions and adverse effects of drugs described in drug monographs were taken into account. However, the physician's time is limited during consultations, and this information is often not consulted. We describe here the design of "Mister VCM", a graphical interface based on the VCM graphical language, facilitating access to drug monographs. We also provide an assessment of the usability of this interface.</p> <p>Methods</p> <p>The "Mister VCM" interface was designed by dividing the screen into two parts: a graphical interactive one including VCM icons and synthetizing drug properties, a textual one presenting on demand drug monograph excerpts. The interface was evaluated over 11 volunteer general practitioners, trained in the use of "Mister VCM". They were asked to answer clinical questions related to fictitious randomly generated drug monographs, using a textual interface or "Mister VCM". When answering the questions, correctness of the responses and response time were recorded.</p> <p>Results</p> <p>"Mister VCM" is an interactive interface that displays VCM icons organized around an anatomical diagram of the human body with additional mental, etiological and physiological areas. Textual excerpts of the drug monograph can be displayed by clicking on the VCM icons. The interface can explicitly represent information implicit in the drug monograph, such as the absence of a given contraindication. Physicians made fewer errors with "Mister VCM" than with text (factor of 1.7; <it>p </it>= 0.034) and responded to questions 2.2 times faster (<it>p </it>< 0.001). The time gain with "Mister VCM" was greater for long monographs and questions with implicit replies.</p> <p>Conclusion</p> <p>"Mister VCM" seems to be a promising interface for accessing drug monographs. Similar interfaces could be developed for other medical domains, such as electronic patient records.</p

Seasonal Influenza Vaccine and Protection against Pandemic (H1N1) 2009-Associated Illness among US Military Personnel

INTRODUCTION: A novel A/H1N1 virus is the cause of the present influenza pandemic; vaccination is a key countermeasure, however, few data assessing prior seasonal vaccine effectiveness (VE) against the pandemic strain of H1N1 (pH1N1) virus are available. MATERIALS AND METHODS: Surveillance of influenza-related medical encounter data of active duty military service members stationed in the United States during the period of April-October 2009 with comparison of pH1N1-confirmed cases and location and date-matched controls. Crude odds ratios (OR) and VE estimates for immunized versus non-immunized were calculated as well as adjusted OR (AOR) controlling for sex, age group, and history of prior influenza vaccination. Separate stratified VE analyses by vaccine type (trivalent inactivated [TIV] or live attenuated [LAIV]), age groups and hospitalization status were also performed. For the period of April 20 to October 15, 2009, a total of 1,205 cases of pH1N1-confirmed cases were reported, 966 (80%) among males and over one-half (58%) under 25 years of age. Overall VE for service members was found to be 45% (95% CI, 33 to 55%). Immunization with prior season's TIV (VE = 44%, 95% CI, 32 to 54%) as well as LAIV (VE = 24%, 95% CI, 6 to 38%) were both found to be associated with protection. Of significance, VE against a severe disease outcome was higher (VE = 62%, 95% CI, 14 to 84%) than against milder outcomes (VE = 42%, 95% CI, 29 to 53%). CONCLUSION: A moderate association with protection against clinically apparent, laboratory-confirmed Pandemic (H1N1) 2009-associated illness was found for immunization with either TIV or LAIV 2008-09 seasonal influenza vaccines. This association with protection was found to be especially apparent for severe disease as compared to milder outcome, as well as in the youngest and older populations. Prior vaccination with seasonal influenza vaccines in 2004-08 was also independently associated with protection