Metalloendopeptidase ADAM-like Decysin 1 (ADAMDEC1) in Colonic Subepithelial PDGFRα+ Cells Is a New Marker for Inflammatory Bowel Disease

Metalloendopeptidase ADAM-Like Decysin 1 (ADAMDEC1) is an anti-inflammatory peptidase that is almost exclusively expressed in the gastrointestinal (GI) tract. We have recently found abundant and selective expression of Adamdec1 in colonic mucosal PDGFRα⁺ cells. However, the cellular origin for this gene expression is controversial as it is also known to be expressed in intestinal macrophages. We found that Adamdec1 mRNAs were selectively expressed in colonic mucosal subepithelial PDGFRα⁺ cells. ADAMDEC1 protein was mainly released from PDGFRα⁺ cells and accumulated in the mucosal layer lamina propria space near the epithelial basement membrane. PDGFRα⁺ cells significantly overexpressed Adamdec1 mRNAs and protein in DSS-induced colitis mice. Adamdec1 was predominantly expressed in CD45⁻ PDGFRα⁺ cells in DSS-induced colitis mice, with only minimal expression in CD45⁺ CD64⁺ macrophages. Additionally, overexpression of both ADAMDEC1 mRNA and protein was consistently observed in PDGFRα⁺ cells, but not in CD64⁺ macrophages found in human colonic mucosal tissue affected by Crohn’s disease. In summary, PDGFRα⁺ cells selectively express ADAMDEC1, which is localized to the colon mucosa layer. ADAMDEC1 expression significantly increases in DSS-induced colitis affected mice and Crohn’s disease affected human tissue, suggesting that this gene can serve as a diagnostic and/or therapeutic target for intestinal inflammation and Crohn’s disease.     

Efficient Total Chemical Synthesis of 13C=18O Isotopomers of Human Insulin for Isotope‐Edited FTIR

Isotope‐edited two‐dimensional Fourier transform infrared spectroscopy (2 D FTIR) can potentially provide a unique probe of protein structure and dynamics. However, general methods for the site‐specific incorporation of stable ¹³C=¹⁸O labels into the polypeptide backbone of the protein molecule have not yet been established. Here we describe, as a prototype for the incorporation of specific arrays of isotope labels, the total chemical synthesis—via a key ester insulin intermediate—of 97 % enriched [(1‐¹³C=¹⁸O)Phe^B24] human insulin: stable‐isotope labeled at a single backbone amide carbonyl. The amino acid sequence as well as the positions of the disulfide bonds and the correctly folded structure were unambiguously confirmed by the X‐ray crystal structure of the synthetic protein molecule. In vitro assays of the isotope labeled [(1‐¹³C=¹⁸O)Phe^B24] human insulin showed that it had full insulin receptor binding activity. Linear and 2 D IR spectra revealed a distinct red‐shifted amide I carbonyl band peak at 1595 cm^?1 resulting from the (1‐¹³C=¹⁸O)Phe^B24 backbone label. This work illustrates the utility of chemical synthesis to enable the application of advanced physical methods for the elucidation of the molecular basis of protein function.     

Assessing genomic prediction accuracy for Holstein sires using bootstrap aggregation sampling and leave-one-out cross validation

Since the introduction of genome-enabled prediction for dairy cattle in 2009, genomic selection has markedly changed many aspects of the dairy genetics industry and enhanced the rate of response to selection for most economically important traits. Young dairy bulls are genotyped to obtain their genomic predicted transmitting ability (GPTA) and reliability (REL) values. These GPTA are a main factor in most purchasing, marketing, and culling decisions until bulls reach 5 yr of age and their milk-recorded offspring become available. At that time, daughter yield deviations (DYD) can be compared with the GPTA computed several years earlier. For most bulls, the DYD align well with the initial predictions. However, for some bulls, the difference between DYD and corresponding GPTA is quite large, and published REL are of limited value in identifying such bulls. A method of bootstrap aggregation sampling (bagging) using genomic BLUP (GBLUP) was applied to predict the GPTA of 2,963, 2,963, and 2,803 young Holstein bulls for protein yield, somatic cell score, and daughter pregnancy rate (DPR), respectively. For each trait, 50 bootstrap samples from a reference population comprising 2011 DYD of 8,610, 8,405, and 7,945 older Holstein bulls were used. Leave-one-out cross validation was also performed to assess prediction accuracy when removing specific bulls from the reference population. The main objectives of this study were (1) to assess the extent to which current REL values and alternative measures of variability, such as the bootstrap standard deviation (SD) of predictions, could detect bulls whose daughter performance deviates significantly from early genomic predictions, and (2) to identify factors associated with the reference population that inform about inaccurate genomic predictions. The SD of bootstrap predictions was a mildly useful metric for identifying bulls whose future daughter performance may deviate significantly from early GPTA for protein and DPR. Leave-one-out cross validation allowed us to identify groups of reference population bulls that were influential on other reference population bulls for protein yield and observe their effects on predictions of testing set bulls, as a whole and individually.     

The sulphidation of alloys used in a fluidized bed combustor

The results of an investigation of the sulphidation behaviour of lncoloy BOOH and AISI 310 stainless steel in coal combustion conditions at temperatures between 650°C and 900°C are presented. Laboratory tests were carried out in controlled environments in the presence of solids representative of fluidized bed combustor operation. The morphology and composition of scale formed during sulphidation was examined and compared with that from tube samples removed from test rigs operated by the National Coal Board. The results have shown that at 900°C a gas mixture corresponding to the equilibrium conditions close to the coexistence point for CaS0₄, CaO and CaS on the phase stability diagram is sufficient to induce sulphidation. However, at lower temperatures an interaction between the deposit and the protective oxide is necessary and in some cases the presence of typical contaminants found in coals is an added requirement. The morphology of the corrosion products found in practice can be reproduced under controlled conditions in laboratory tests. The results are interpreted in terms of likely mechanisms of the processes involved.     

Localization of trehalose in partially hydrated DOPC bilayers: insights into cryoprotective mechanisms

Trehalose, a natural disaccharide with bioprotective properties, is widely recognized for its ability to preserve biological membranes during freezing and dehydration events. Despite debate over the molecular mechanisms by which this is achieved, and that different mechanisms imply quite different distributions of trehalose molecules with respect to the bilayer, there are no direct experimental data describing the location of trehalose within lipid bilayer membrane systems during dehydration. Here, we use neutron membrane diffraction to conclusively show that the trehalose distribution in a dioleoylphosphatidylcholine (DOPC) system follows a Gaussian profile centred in the water layer between bilayers. The absence of any preference for localizing near the lipid headgroups of the bilayers indicates that the bioprotective effects of trehalose at physiologically relevant concentrations are the result of non-specific mechanisms that do not rely on direct interactions with the lipid headgroups.     

Detection of Non-metallic Inclusions in Steel Continuous Casting Billets

This work applied automated particle analysis to study non-metallic inclusions in steel. Compared with traditional methods, the approach has the advantage of capturing the morphology, measuring the size, recording the original positions, and identifying the composition of inclusions on a selected area in a short time. The morphology and composition of typical inclusions were analyzed using partial acid extraction and discussed through thermodynamic calculation. Steel samples were collected from the entire cross section of billets cast during times of steady state and ladle change. The spatial distribution of inclusions agreed well with the measurement of the total oxygen. The spatial distribution of inclusions was plotted to represent the entrapment positions of inclusions on the casting strand and their concentration on the cross section of the billet. Also, regarding the different size and type of inclusions, the spatial distribution of classified inclusions was explored such as the distribution of sulfide, oxide, and high sodium and potassium content inclusions. The sufficient information could be used to identify the source of inclusions and guide the steel refining process.     

Use of Self-assembled Monolayers at Variable Coverage to Control Interface Bonding in a Model Study of Interfacial Fracture: Pure Shear Loading

The relationships between fundamental interfacial interactions, energy dissipation mechanisms, and fracture stress or fracture energy in a glassy thermoset/inorganic solid joint are not well understood. This subject is addressed with a model system involving an epoxy adhesive on a polished silicon wafer containing its native oxide. The proportions of physical and chemical interactions at the interface, and the in-plane distribution, are varied using self-assembling monolayers of octadecyltrichlorosilane (ODTS). The epoxy interacts strongly with the bare silicon oxide surface, but interacts only weakly with the methylated tails of the ODTS monolayer. The fracture stress is examined as a function of ODTS coverage in the napkin-ring (nominally pure shear) loading geometry. The relationship between fracture stress and ODTS coverage is catastrophic, with a large change in fracture stress occurring over a narrow range of ODTS coverage. This transition in fracture stress does not correspond to a wetting transition of the epoxy. Rather, the transition in fracture stress corresponds to the onset of large-scale plastic deformation within the epoxy. We postulate that the transition in fracture stress occurs when the local stress that the interface can support becomes comparable to the yield stress of the epoxy. The fracture results are independent of whether the ODTS deposition occurs by island growth (T_dep = 10°C) or by homogeneous growth (T_dep = 24°C).