Detectability of the degree of freeze damage in meat analytic-tool depends on selection

Novel freezing solutions are constantly being developed to reduce quality loss in meat production chains. However, there is limited focus on identifying the sensitive analytical tools needed to directly validate product changes that result from potential improvements in freezing technology. To benchmark analytical tools relevant to meat research and production, we froze pork samples using traditional (−25 °C, −35 °C) and cryogenic freezing (−196 °C). Three classes of analyses were tested for their capacity to separate different freeze treatments: thaw loss testing, bioelectrical spectroscopy (nuclear magnetic resonance, microwave, bioimpedance) and low-temperature microscopy (cryo-SEM). A general effect of freeze treatment was detected with all bioelectrical methods. Yet, only cryo-SEM resolved quality differences between all freeze treatments, not only between cryogenic and traditional freezing. The detection sensitivity with cryo-SEM may be explained by testing meat directly in the frozen state without prior defrosting. We discuss advantages, shortcomings and cost factors in using analytical tools for quality monitoring in the meat sector

Orphan receptor GPR110, an oncogene overexpressed in lung and prostate cancer

<p>Abstract</p> <p>Background</p> <p>GPR110 is an orphan G protein-coupled receptor--a receptor without a known ligand, a known signaling pathway, or a known function. Despite the lack of information, one can assume that orphan receptors have important biological roles. In a retroviral insertion mutagenesis screen in the mouse, we identified GPR110 as an oncogene. This prompted us to study the potential isoforms that can be gleaned from known GPR110 transcripts, and the expression of these isoforms in normal and transformed human tissues.</p> <p>Methods</p> <p>Various epitope-tagged isoforms of GPR110 were expressed in cell lines and assayed by western blotting to determine cleavage, surface localization, and secretion patterns. GPR110 transcript and protein levels were measured in lung and prostate cancer cell lines and clinical samples, respectively, by quantitative PCR and immunohistochemistry.</p> <p>Results</p> <p>We found four potential splice variants of GPR110. Of these variants, we confirmed three as being expressed as proteins on the cell surface. Isoform 1 is the canonical form, with a molecular mass of about 100 kD. Isoforms 2 and 3 are truncated products of isoform 1, and are 25 and 23 kD, respectively. These truncated isoforms lack the seven-span transmembrane domain characteristic of GPR proteins and thus are not likely to be membrane anchored; indeed, isoform 2 can be secreted. Compared with the median gene expression of ~200 selected genes, GPR110 expression was low in most tissues. However, it had higher than average gene expression in normal kidney tissue and in prostate tissues originating from older donors. Although identified as an oncogene in murine T lymphomas, GPR110 is greatly overexpressed in human lung and prostate cancers. As detected by immunohistochemistry, GPR110 was overexpressed in 20 of 27 (74%) lung adenocarcinoma tissue cores and in 17 of 29 (59%) prostate adenocarcinoma tissue cores. Additionally, staining with a GPR110 antibody enabled us to differentiate between benign prostate hyperplasia and potential incipient malignancy.</p> <p>Conclusion</p> <p>Our work suggests a role for GPR110 in tumor physiology and supports it as a potential therapeutic candidate and disease marker for both lung and prostate cancer.</p

Capturing, sharing and analysing biophysical data from protein engineering and protein characterization studies

Large amounts of data are being generated annually on the connection between the sequence, structure and function of proteins using site-directed mutagenesis, protein design and directed evolution techniques. These data provide the fundamental building blocks for our understanding of protein function, molecular biology and living organisms in general. However, much experimental data are never deposited in databases and is thus ‘lost’ in journal publications or in PhD theses. At the same time theoretical scientists are in need of large amounts of experimental data for benchmarking and calibrating novel predictive algorithms, and theoretical progress is therefore often hampered by the lack of suitable data to validate or disprove a theoretical assumption. We present PEAT (Protein Engineering Analysis Tool), an application that integrates data deposition, storage and analysis for researchers carrying out protein engineering projects or biophysical characterization of proteins. PEAT contains modules for DNA sequence manipulation, primer design, fitting of biophysical characterization data (enzyme kinetics, circular dichroism spectroscopy, NMR titration data, etc.), and facilitates sharing of experimental data and analyses for a typical university-based research group. PEAT is freely available to academic researchers at http://enzyme.ucd.ie/PEAT

The Adhesion GPCR GPR125 is specifically expressed in the choroid plexus and is upregulated following brain injury

<p>Abstract</p> <p>Background</p> <p>GPR125 belongs to the family of <it>Adhesion </it>G protein-coupled receptors (GPCRs). A single copy of GPR125 was found in many vertebrate genomes. We also identified a <it>Drosophila </it>sequence, DmCG15744, which shares a common ancestor with the entire Group III of <it>Adhesio</it>n GPCRs, and also contains Ig, LRR and HBD domains which were observed in mammalian GPR125.</p> <p>Results</p> <p>We found specific expression of GPR125 in cells of the choroid plexus using <it>in situ </it>hybridization and protein-specific antibodies and combined <it>in situ</it>/immunohistochemistry co-localization using cytokeratin, a marker specific for epithelial cells. Induction of inflammation by LPS did not change GPR125 expression. However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.</p> <p>Conclusion</p> <p>These findings suggest that GPR125 plays a functional role in choroidal and hippocampal response to injury.</p

Control of interneuron dendritic growth through NRG1/erbB4-mediated kalirin-7 disinhibition.

Neuregulin 1 (NRG1) is a secreted trophic factor that activates the postsynaptic erbB4 receptor tyrosine kinase. Both NRG1 and erbB4 have been repeatedly associated with schizophrenia, but their downstream targets are not well characterized. ErbB4 is highly abundant in interneurons, and NRG1-mediated erbB4 activation has been shown to modulate interneuron function, but the role for NRG1-erbB4 signaling in regulating interneuron dendritic growth is not well understood. Here we show that NRG1/erbB4 promote the growth of dendrites in mature interneurons through kalirin, a major dendritic Rac1-GEF. Recent studies have shown associations of the KALRN gene with schizophrenia. Our data point to an essential role of phosphorylation in kalirin-7's C terminus as the critical site for these effects. As reduced interneuron dendrite length occurs in schizophrenia, understanding how NRG1-erbB4 signaling modulates interneuron dendritic morphogenesis might shed light on disease-related alterations in cortical circuits

The impact of change in a doctor's job position: a five-year cohort study of job satisfaction among Norwegian doctors

<p>Abstract</p> <p>Background</p> <p>Job satisfaction among physicians may be of importance to their individual careers and their work with patients. We lack prospective studies on whether a change in a doctor's job position influences their job satisfaction over a five-year period if we control for other workload factors.</p> <p>Methods</p> <p>A longitudinal national cohort of all physicians who graduated in Norway in 1993 and 1994 was surveyed by postal questionnaire in 2003 (T1) and 2008 (T2). Outcomes were measured with a 10-item job satisfaction scale. Predictor variables in a multiple regression model were: change in job position, reduction in work-home interface stress, reduction in work hours, age, and gender.</p> <p>Results</p> <p>A total of 59% of subjects (306/522) responded at both time points. The mean value of job satisfaction in the total sample increased from 51.6 (SD = 9.0) at T1 to 53.4 (SD = 8.2) at T2 (paired <it>t </it>test, <it>t </it>= 3.8, <it>p </it>< 0.001). The major groups or positions at T1 were senior house officers (45%), chief specialists in hospitals (23%), and general practitioners (17%), and the latter showed the highest levels of job satisfaction. Physicians who changed position during the period (n = 176) experienced an increase in job satisfaction from 49.5 (SD = 8.4) in 2003 to 52.9 (SD = 7.5) in 2008 (paired <it>t </it>test, <it>t </it>= 5.2, <it>p </it>< 0.001). Job satisfaction remained unchanged for physicians who stayed in the same position. There was also an increase in satisfaction among those who changed from positions other than senior house officer at T1 (<it>p </it>< 0.01). The significant adjusted predictor variables in the multiple regression model were the change in position from senior house officer at T1 to any other position (β = 2.83, <it>p </it>< 0.001), any change in job position (from any position except SHO at T1) (β = 4.18, <it>p </it>< 0.01) and reduction in work-home interface stress (β = 1.04, <it>p </it>< 0.001).</p> <p>Conclusions</p> <p>The physicians experienced an increase in job satisfaction over a five-year period, which was predicted by a change in job position and a reduction in work-home stress. This study has implications with respect to career advice for young doctors.</p

Molecular evolution of a chordate specific family of G protein-coupled receptors

<p>Abstract</p> <p>Background</p> <p>Chordate evolution is a history of innovations that is marked by physical and behavioral specializations, which led to the development of a variety of forms from a single ancestral group. Among other important characteristics, vertebrates obtained a well developed brain, anterior sensory structures, a closed circulatory system and gills or lungs as blood oxygenation systems. The duplication of pre-existing genes had profound evolutionary implications for the developmental complexity in vertebrates, since mutations modifying the function of a duplicated protein can lead to novel functions, improving the evolutionary success.</p> <p>Results</p> <p>We analyzed here the evolution of the GPRC5 family of G protein-coupled receptors by comprehensive similarity searches and found that the receptors are only present in chordates and that the size of the receptor family expanded, likely due to genome duplication events in the early history of vertebrate evolution. We propose that a single GPRC5 receptor coding gene originated in a stem chordate ancestor and gave rise by duplication events to a gene family comprising three receptor types (GPRC5A-C) in vertebrates, and a fourth homologue present only in mammals (GPRC5D). Additional duplications of GPRC5B and GPRC5C sequences occurred in teleost fishes. The finding that the expression patterns of the receptors are evolutionarily conserved indicates an important biological function of these receptors. Moreover, we found that expression of GPRC5B is regulated by vitamin A <it>in vivo</it>, confirming previous findings that linked receptor expression to retinoic acid levels in tumor cell lines and strengthening the link between the receptor expression and the development of a complex nervous system in chordates, known to be dependent on retinoic acid signaling.</p> <p>Conclusions</p> <p>GPRC5 receptors, a class of G protein-coupled receptors with unique sequence characteristics, may represent a molecular novelty that helped non-chordates to become chordates.</p

Comparative Coastal Risk Index (CCRI): A multidisciplinary risk index for Latin America and the Caribbean

As the world's population grows to a projected 11.2 billion by 2100, the number of people living in low-lying areas exposed to coastal hazards is projected to increase. Critical infrastructure and valuable assets continue to be placed in vulnerable areas, and in recent years, millions of people have been displaced by natural hazards. Impacts from coastal hazards depend on the number of people, value of assets, and presence of critical resources in harm's way. Risks related to natural hazards are determined by a complex interaction between physical hazards, the vulnerability of a society or social-ecological system and its exposure to such hazards. Moreover, these risks are amplified by challenging socioeconomic dynamics, including poorly planned urban development, income inequality, and poverty. This study employs a combination of machine learning clustering techniques (Self Organizing Maps and K-Means) and a spatial index, to assess coastal risks in Latin America and the Caribbean (LAC) on a comparative scale. The proposed method meets multiple objectives, including the identification of hotspots and key drivers of coastal risk, and the ability to process large-volume multidimensional and multivariate datasets, effectively reducing sixteen variables related to coastal hazards, geographic exposure, and socioeconomic vulnerability, into a single index. Our results demonstrate that in LAC, more than 500,000 people live in areas where coastal hazards, exposure (of people, assets and ecosystems) and poverty converge, creating the ideal conditions for a perfect storm. Hotspot locations of coastal risk, identified by the proposed Comparative Coastal Risk Index (CCRI), contain more than 300,00 people and include: El Oro, Ecuador; Sinaloa, Mexico; Usulutan, El Salvador; and Chiapas, Mexico. Our results provide important insights into potential adaptation alternatives that could reduce the impacts of future hazards. Effective adaptation options must not only focus on developing coastal defenses, but also on improving practices and policies related to urban development, agricultural land use, and conservation, as well as ameliorating socioeconomic conditions

Prion Protein Is a Key Determinant of Alcohol Sensitivity through the Modulation of N-Methyl-D-Aspartate Receptor (NMDAR) Activity

The prion protein (PrP) is absolutely required for the development of prion diseases; nevertheless, its physiological functions in the central nervous system remain elusive. Using a combination of behavioral, electrophysiological and biochemical approaches in transgenic mouse models, we provide strong evidence for a crucial role of PrP in alcohol sensitivity. Indeed, PrP knock out (PrP−/−) mice presented a greater sensitivity to the sedative effects of EtOH compared to wild-type (wt) control mice. Conversely, compared to wt mice, those over-expressing mouse, human or hamster PrP genes presented a relative insensitivity to ethanol-induced sedation. An acute tolerance (i.e. reversion) to ethanol inhibition of N-methyl-D-aspartate (NMDA) receptor-mediated excitatory post-synaptic potentials in hippocampal slices developed slower in PrP−/− mice than in wt mice. We show that PrP is required to induce acute tolerance to ethanol by activating a Src-protein tyrosine kinase-dependent intracellular signaling pathway. In an attempt to decipher the molecular mechanisms underlying PrP-dependent ethanol effect, we looked for changes in lipid raft features in hippocampus of ethanol-treated wt mice compared to PrP−/− mice. Ethanol induced rapid and transient changes of buoyancy of lipid raft-associated proteins in hippocampus of wt but not PrP−/− mice suggesting a possible mechanistic link for PrP-dependent signal transduction. Together, our results reveal a hitherto unknown physiological role of PrP on the regulation of NMDAR activity and highlight its crucial role in synaptic functions