COL11A1 is associated with developmental dysplasia of the hip and secondary osteoarthritis in the HUNT study.

Objective: Developmental dysplasia of the hip (DDH) is a congenital condition affecting 2-3% of all infants. DDH increases the risk of osteoarthritis, is the cause of 30 â€‹% of all total hip arthroplasties (THAs) in adults <40 years of age and can result in loss of life quality. Our aim was to explore the genetic background of DDH in order to improve diagnosis, management and longterm outcome. Design: We used the large, ongoing, longitudinal Trøndelag Health Study (HUNT) database. Case definition was based on ICD-9/-10 diagnoses of DDH, or osteoarthritis secondary to DDH. Analyses were performed using SAIGE software, with covariates including sex, batch, birth year and principal components. We included only single nucleotide polymorphisms (SNPs) with minor allele frequency (MAF) ≥ 0.01, R2≥ 0.8 and Hardy-Weinberg equilibrium (HWE) P-value ≥ 0.0001. Significance level was set at p â€‹< â€‹5 â€‹× â€‹10-8. Meta-analysis using data from DDH and primary osteoarthritis genome-wide association studies (GWASs) was done using METAL software. The study was approved by the regional ethical committee. Results: Analysis included 69,500 individuals, of which 408 cases, and 8,531,386 SNPs. Two SNPs near COL11A1 were significantly associated with DDH; rs713162 (ߠ​= â€‹-0.43, SE â€‹= â€‹0.07, p â€‹= â€‹8.4 â€‹× â€‹10-9) and rs6577334 (ߠ​= â€‹-0.43, SE â€‹= â€‹0.08, p â€‹= â€‹8.9 â€‹× â€‹10-9). COL11A1 has previously been associated with acetabular dysplasia and osteoarthritis. Meta-analysis supported previous GWAS findings of both DDH and primary osteoarthritis. Conclusions: This large, genome-wide case-control study indicates an association between COL11A1 and DDH and is an important contribution to investigating the etiology of DDH, with further research needed.

Evaluation of biological air filters for livestock ventilation air by membrane inlet mass spectrometry.

Biological air filters have been proposed as a cost-effective technology for reducing odor emissions from intensive swine production facilities. In this work we present results from the application of membrane inlet mass spectrometry (MIMS) for continuously monitoring the removal of odorous compounds in biological air filters. The sensitivity and selectivity were tested on synthetic samples of selected odorous compounds, and linearity and detection limits in the lower ppb range were demonstrated for all compounds tested (methanethiol, dimethyl sulfide, carboxylic acids, 4-methylphenol, aldehydes, indole, and skatole) except trimethylamine. The method was applied in situ at two full-scale filters installed at swine houses. The results have been compared with analyses by thermal desorption gas chromatography-mass spectrometry (TD-GC/MS), and odor was measured by olfactometry. By comparison with TD-GC/MS, observed MIMS signals were assigned to 4-methylphenol, 4-ethylphenol, indole, skatole, the sum of volatile reduced organic sulfur compounds (ROS), and three subgroups of carboxylic acids. The removal rates were observed to be related to air-water partitioning with removal efficiencies in the range of 0 to 50% for low-soluble organic sulfur compounds and high removal efficiencies (typically 80-100%) for more soluble phenols and carboxylic acids. Based on the results and published odor threshold values, it is estimated that the low removal efficiency of ROS is the main limitation for achieving a higher odor reduction.

Comparison of glucose, glucose 6-phosphate, ribose, and mannose as flavour precursors in pork; the effect of monosaccharide addition on flavour generation.

The effect of glucose, glucose 6-phosphate, mannose and ribose on the generation of aroma volatiles in pork was investigated. The monosaccharides were added individually to minced pork prior to heat treatment (160°C for 10min) in the following concentrations: glucose (27.5µmol/g), ribose (1.2µmol/g), mannose (8.3µmol/g) and glucose 6-phosphate (0.5µmol/g). The natural concentrations of the monosaccharides in the pork used were found to be 4.0µmol/g for glucose, 0.1µmol/g for ribose, 0.3µmol/g for mannose and 2.6µmol/g for glucose 6-phosphate. The major aroma compounds identified in the headspace of the heated samples were pyrazines, aldehydes (Strecker and lipid-derived), ketones, and sulphides. Glucose generated the highest amounts of volatiles followed by glucose 6-phosphate. However, when related to the added concentration of glucose 6-phosphate, this phosphorylated monosaccharide showed the highest aroma generating potential. The addition of ribose did not increase the concentration of volatiles compared with pork without the added monosaccharide. The fates of ribose 5-phosphate and ribose in pork were studied over time. The concentrations of ribose and ribose 5-phosphate clearly decreased during 2h equilibration, which may be due to enzymatic activities. These precursors may, therefore, be less important pork flavour precursors than glucose and glucose 6-phosphate.

Accelerated transport of (90)Sr following a release of high ionic strength solution in vadose zone sediments.

Numerical simulation of cation exchange and mineral precipitation/dissolution reactions using the multiphase reactive geochemical transport code TOUGHREACT has provided important insight into the distribution of (90)Sr among layers of geologic strata in a complex vadose zone at the U. S. Department of Energy's Idaho National Laboratory. During a transfer operation in November 1972, 70.4 m(3) of acidic, high ionic strength liquid containing 15,900 Ci of (90)Sr was released over five days into alluvial gravels 137 m above the Snake River Plain Aquifer. Sampling data from perched water zones 33 m below the release contain very high levels of (90)Sr as do soil samples obtained nearer the point of release. Use of traditional simulation approaches using laboratory-measured constant partitioning coefficients (K(d)) cannot simultaneously explain perched water and soil concentrations. To address the discrepancy, a reactive transport approach was adopted to include competitive cation exchange, dissolution/precipitation of calcite, carbon dioxide gas production and transport, and gibbsite precipitation. Simulation results using this model suggest that some of the (90)Sr could have been transported very rapidly immediately after the release with the acceleration facilitated by competition for cation exchange sites with high sodium concentrations in the released liquid and calcium dissolved from calcite, and to a lesser extent by formation of aqueous complexes with nitrate. Once the leading edge of the liquid assemblage was flushed from the alluvium, the mobility of the remaining (90)Sr decreased significantly in the absence of the competing cations. Calculations indicate that there should be a net increase in calcite, suggesting that (90)Sr could be entrained in the mineral lattice, but insufficient field data exist for confirmation. Sensitivity studies show that the cation exchange selectivity coefficients were the most sensitive individual parameters determining the (90)Sr distribution. However, the most sensitive overall quantity was the total ion exchange capacity which is a function of the moles of exchange sites per volume of pore water, the cation exchange capacity, and the total volume wetted by the infiltrating solution. In contrast, the future mobility of (90)Sr was found to be relatively insensitive to the normal range in the composition of influxing precipitation and anthropogenic waters.

Physical changes of significance for early post mortem water distribution in porcine M. longissimus.

The post mortem changes in water mobility and distribution were followed in porcine muscle (M. longissimus dorsi) samples using continuous low-field NMR relaxation measurements and simultaneous measurement of changes in muscle impedance as an indirect measure of membrane integrity as well as muscle contraction measurements using a rigormeter instrument. Distributed exponential fitting analysis of NMR T(2) relaxation data revealed the presence of three distinct water populations (T(20), T(21), T(22)) within the muscle during its conversion to meat. Comparison of T(2) relaxation patterns and contraction data indicates that rigor development affects the attributes of the T(21) water population and thereby contributes to myofibrillar water characteristics post mortem, as the T(21) water population is believed to reflect inter/intra-myofibrillar water. The volume of the water population believed to reflect extra-cellular water (T(22)) in the living muscle. Early post mortem T(22) decreased slightly within the first 2-3 h post mortem followed by an increase and a change in its characteristic time constant. This was ascribed to an initial muscle cell swelling followed by water being expelled from the cellular space into the extra-myofibrillar space. Comparison of changes in the T(22) water population and impedance characteristics within the muscle during its conversion to meat revealed close relationship between progresses in the two attributes. Obtained data strongly support that the post mortem reorganization of water is closely associated with membrane properties, which moreover was found to affect the final water-holding capacity of the meat. Finally, a model for early post mortem events leading to changes in the distribution of water within muscles is proposed.

Asynchronous cell cycle and asymmetric vacuolar inheritance in true hyphae of Candida albicans.

Candida albicans forms unconstricted hyphae in serum-containing medium that are divided into discrete compartments. Time-lapse photomicroscopy, flow cytometry, and a novel three-dimensional imaging system were used to demonstrate that the kinetics and cell cycle events accompanying hyphal development were correlated with dynamic changes in vacuole morphology and the pattern of vacuole inheritance. Apical cells of hyphae underwent continuous extension before and after the first cytokinesis event. However, the resulting mother cell and sub-apical compartments did not immediately reenter the cell cycle and instead underwent cell cycle arrest before reentering the cycle. Vacuole was inherited asymmetrically at cytokinesis so that the distal, arrested compartments inherited most vacuole and the growing apical cell inherited most cytoplasm. Hydroxyurea release experiments demonstrated that the arrested, vacuolated hyphal compartments were in the G(1) phase of the cycle. The period of cell cycle arrest was decreased by the provision of assimilatable forms of nitrogen, suggesting that the hyphal cell cycle is regulated by nitrogen limitation that results in sup-apical cell cycle arrest. This pattern of growth is distinct from that of the synchronous, symmetrical development of pseudohyphae of C. albicans and other yeast species. These observations suggest that the cellular vacuole space correlates with alterations in the cell cycles of different cell types and that the total organelle space may influence size-regulated functions and hence the timing of the eukaryotic cell cycle.

A highly specific system for efficient enzymatic removal of tags from recombinant proteins.

The TAGZyme system allows efficient and precise exoproteolytic cleavage of N-terminal affinity tags, such as the 6xHis tag, from recombinant proteins. In combination with Ni-NTA technology, the TAGZyme system provides high-purity proteins free of vector-encoded amino acids for use in applications that demand recombinant reagents, an absence of nonspecific cleavage, and a complete removal of all impurities from the target protein preparation. We present results of recent studies on the use of the TAGZyme system. The N-terminal affinity tag sequences encoded by the TAGZyme pQE expression vectors are optimized with respect to their cleavage using TAGZyme exoproteases. The vectors have multiple cloning site sequences designed to allow complete exoproteolytic removal of the encoded N-terminal affinity tag regardless of the restriction site used for cloning. The efficient cleavage reaction and removal of the TAGZyme enzymes by subtractive Ni-NTA chromatography is demonstrated for 6xHis-interleukin-1beta and 6xHis-TNF. In both cases, more than 99.8% of TAGZyme proteolytic activity was separated from recovered, detagged proteins. N-terminal analyses by Edman degradation revealed the predicted sequences of the native proteins and indicated a purity in excess of 99%. For cleavage of both 6xHis-tagged GFP and IL-1beta, DAPase enzyme gave an average cleavage rate of 1.5 +/- 0.5 min per amino acid.

Physiological and structural events post mortem of importance for drip loss in pork.

Early post mortem metabolism and structural changes from 3 to 24 h, together with pH, temperature and impedance Py development were investigated in 37 Duroc×Landrace×Large White (DLY) pigs covering a range of drip loss from 2.2 to 12.6%. Multivariate statistical analysis was used to assess the impacts of different metabolites, pH and temperature, impedance, cytoskeletal protein degradation and extracellular cross-sectional area on drip loss. Taken as single factors, the concentration of lactate could explain 80% of the variation in drip, inosine monophosphate (IMP) and adenosine triphosphate (ATP) concentration explained 71 and 68%, respectively, whereas inosin and glycogen levels explained only 59 and 60%. The extracellular area was found to explain 39% of the variation in drip. The area between fibres provided more significant information than did the area between fibre bundles. The degradation of the cytoskeletal proteins was not related to drip loss. Impedance Py development over 24 h could explain 66% of the variation in drip, whereas pH and temperature explained 85 and 87%, respectively. A model including all measured variables could explain 83% of the variation in drip. However, only pH, temperature, impedance, [ATP](1 h) and [lactate](1 h and 2 h) were significant in relation to drip. By reducing the variables in the multivariate analysis, 89% of the variation in drip could be explained by a model containing only pH(2 h) and temperature(1 min). To explain variation in drip loss, pH and temperature measurements at significant time points were sufficient. Variation in post-mortem metabolites did, however, explain why variation in pH and temperature occurred. Development of drip channels was ruled by pH and temperature while impedance development was highly correlated to pH. This multi-faceted study shows those parameters, which can best be used to indicate or predict WHC, as well as those indicating the basic mechanism underlying variations in drip.