REGULATION OF THE BIOAVAILABILITY OF THIOREDOXIN IN THE LENS BY A SPECIFIC THIOREDOXIN-BINDING PROTEIN (TBP-2)

Thioredoxin (TRx) is known to control redox homeostasis in cells. In recent years, a specific TRx binding protein called thioredoxin binding protein-2 (TBP-2) was found in other cell types and it appeared to negatively regulate TRx bioavailability and thereby control TRx biological function. In view of the sensitivity of lens transparency to redox status, proper regulation of TRx bioavailability is of the utmost importance. This study was conducted to examine the presence and function of TBP-2 in human lens epithelial cells (HLE B3). We cloned human lens TBP-2 from a human cDNA library (GenBank accession number AY 594328) and showed that it is fully homologous to the human brain TBP-2 gene. The recombinant TBP-2 protein was partially purified and mass spectrometric analysis confirmed its sequence homology to that of brain TBP-2. Immunoprecipitates obtained from HLE B3 cells using anti-TRx and anti-TBP-2 antibodies showed the presence of TRx and TBP-2 in immunoprecipitates indicating the formation of a TRx-TBP-2 complex in vivo. Furthermore, under H2O2-stress conditions, TRx gene expression was transiently up-regulated while TBP-2 gene expression was inversely down-regulated as seen in both HLE B3 cells and in the epithelial cell layers from cultured pig lenses. Cells with overexpressed TBP-2 showed lower TRx activity, grew slower and were more susceptible to oxidative stress-induced apoptosis. This is the first report of the presence of a TRx-specific binding protein in the lens. Our data suggest that TBP-2 is likely a negative regulator for the bioavailability, and therefore, the overall function of TRx in the lens

Bayesian Methods for Genomic Prediction and Genome-Wide Association Studies combining Information on Genotyped and Non-Genotyped Individuals

Genomic prediction involves using high-density marker genotypes to characterize the impact on performance of every region of the genome, and using that information to predict performance of genotyped selection candidates. This is a relatively new technology and is now gaining traction in personalized medicine and in various livestock industries. Our new approach promises to overcome serious limitations with existing techniques for genomic prediction

Fine-tuning the predicted position of genes associated with economic traits in livestock

Different methods that estimate the position of a gene on a chromosome were tested in computer-simulated populations to determine their accuracy. Given the same amount of genetic information from the animals, one method performed better than the others. In situations where experimental costs were assumed to be equal but genetic information could vary, this method was no longer the most accurate. Further study of this method found that the animals’ genetic information must be used in a specific way in order to obtain the most accurate position of the gene. These methods will be useful in identifying genes and the genetic differences between animals that can be used for genetic improvement of livestock

Genomic Prediction Using Linkage Disequilibrium and Co-segregation

A linear mixed model fitting both genome-wide cosegregation (CS) and linkage disequilibrium (LD) was developed to improve accuracy of genetic prediction for pedigreed populations of unrelated families that have half sibs represented in both training and validation. Cosegregation was modeled as the effects of genome-wide1-centimorgan haplotypes that one individual inherits from pedigree founders through identity-by-descent, while LD was modeled as allele substitution effects of all marker genotypes. Prediction accuracy of the LD-CS method was compared to the accuracy of three LD methods – GBLUP, BayesA and BayesB, using simulated datasets of varying numbers of paternal half sib families. Results show that the LD-CS method tended to have higher accuracy than any of the LD methods. With an increase in the number of families, the accuracy of the LD-CS method persisted, while the accuracy of the LD methods dropped. The results indicate that by fitting CS explicitly, the LD-CS method has higher and more consistent prediction accuracy than LD methods

Improved Accuracy of Genomic Prediction for Traits with Rare QTL by Fitting Haplotypes

Genomic prediction estimates breeding values by exploiting linkage disequilibrium (LD) between quantitative trait loci (QTL) and single nucleotide polymorphisms (SNPs). High LD cannot occur when QTL and SNPs have different minor allele frequencies (MAF). Marker panels tend to use SNPs with high MAF and will have limited ability to predict rare QTL alleles. In practice, increasing SNP density has not improved prediction accuracy. A possible reason is that many traits are characterized by rare QTL. In that case, linear models fitting haplotypes could have advantage because haplotypes can be in complete LD with QTL alleles. SNP genotypes were simulated to resemble 600K chip for the bovine genome. Genomic breeding values were predicted using either SNP genotypes or non-overlapping haplotypes. When QTL had low MAF, the haplotype model had significantly higher accuracy than the SNP model. Results show that fitting haplotypes can improve the accuracy of genomic prediction for traits controlled by rare QTL

Genomic Selection of Purebred Animals for Crossbred Performance in the Presence of Dominant Gene Action

The primary objective of this study was to assess the performance of different genomic prediction models applied to the selection of purebreds for crossbred performancebased on high-density marker data. Our results suggest that in the presence of dominant gene action, selection based on the dominance model is superior to both the a breed-specific allele model and an additive model in terms of maximizing crossbred performance through purebred selection, especially when training is not updated each generatio

Genome-wide association study of infectious bovine keratoconjunctivitis in Angus cattle

Background Infectious Bovine Keratoconjunctivitis (IBK) in beef cattle, commonly known as pinkeye, is a bacterial disease caused by Moraxella bovis. IBK is characterized by excessive tearing and ulceration of the cornea. Perforation of the cornea may also occur in severe cases. IBK is considered the most important ocular disease in cattle production, due to the decreased growth performance of infected individuals and its subsequent economic effects. IBK is an economically important, lowly heritable categorical disease trait. Mass selection of unaffected animals has not been successful at reducing disease incidence. Genome-wide studies can determine chromosomal regions associated with IBK susceptibility. The objective of the study was to detect single-nucleotide polymorphism (SNP) markers in linkage disequilibrium (LD) with genetic variants associated with IBK in American Angus cattle. ResultsThe proportion of phenotypic variance explained by markers was 0.06 in the whole genome analysis of IBK incidence classified as two, three or nine categories. Whole-genome analysis using any categorisation of (two, three or nine) IBK scores showed that locations on chromosomes 2, 12, 13 and 21 were associated with IBK disease. The genomic locations on chromosomes 13 and 21 overlap with QTLs associated with Bovine spongiform encephalopathy, clinical mastitis or somatic cell count. ConclusionsResults of these genome-wide analyses indicated that if the underlying genetic factors confer not only IBK susceptibility but also IBK severity, treating IBK phenotypes as a two-categorical trait can cause information loss in the genome-wide analysis. These results help our overall understanding of the genetics of IBK and have the potential to provide information for future use in breeding schemes

LOW MOLECULAR WEIGHT PROTEIN TYROSINE PHOSPHATASE (LMW-PTP) AND ITS POSSIBLE PHYSIOLOGICAL FUNCTIONS OF REDOX SIGNALING IN THE EYE LENS

Low molecular weight protein tyrosine phosphatase (LMW-PTP) was cloned from human lens epithelial B3 cells (HLE B3) and the recombinant enzyme was purified to homogeneity. The pure enzyme reacted positively with anti-LMW-PTP antibody, displayed tyrosine-specific phosphatase activity and was extremely sensitive to H2O2. The inactivated LMW-PTP could be regenerated by thioltransferase (TTase)/GSH system as demonstrated by both activity assay and by mass spectrometry (MS). The MS study also showed that an intramolecular disulfide bond was formed between C13 and C18 at the active site, and was reduced by the TTase/GSH system. The putative role of LMW-PTP in regulating platelet derived growth factor (PDGF)-stimulated cell signaling was demonstrated in wild type mouse lens epithelial cells (LEC) in which LMW-PTP was transiently inactivated, corroborated with the transient phosphorylation of Tyr857 at the active site of PDGF receptor and the downstream signaling components of Akt and ERK1/2. In contrast, LMW-PTP activity in PDGF-stimulated LEC from TTase −/− mice was progressively lost, concomitant with the high basal and sustained high phosphorylation levels at Tyr857, Akt and ERK1/2. We conclude that the reversible LMW-PTP activity regulated by ROS-mediated oxidation and TTase/GSH reduction is the likely mechanism of redox signaling in lens epithelial cells

Electroless thin film CoNiFe-B alloys for integrated magnetics on Si

Electroless magnetic thin films have been deposited from borane-based baths suitable for use in integrated magnetics on Si applications. The baths were developed for compatibility with standard photoresist for microfabrication of integrated magnetics on Si. The specific formulations, which differ from those reported previously, yield uniform, high saturation magnetisation (up to 2.15 T) deposits with low coercivity (<2 Oe). The resistivity of the film can be increased to minimise eddy current losses by using higher dimethylamine borane (DMAB) content or the inclusion of a second reducing agent, hypophosphite, to facilitate phosphorus codeposition of up to 7 at.%. The Ni content in the plating bath has been shown to exert significant influence over the composition, deposition rate and coercivity. XRD analysis suggests that the deposits consist of nanocrystalline phase with grains <20 nm. Such small grains are consistent with the observed low coercivity of the deposits