Ovine herpesvirus 2 (OvHV-2) is a gammaherpesvirus that causes sheep-associated malignant catarrhal fever (SA-MCF), a frequently fatal disease mainly of ruminants. This study was designed to define virus-host dynamics following experimental OvHV-2 infection in bison. A transient peak in viral DNA accompanied by the presence of OvHV-2 ORF25, ORF50 and ORF73 transcripts was observed in lungs only from 9 to 12 days post-inoculation (DPI), suggesting occurrence of viral replication. This initial viral replication was associated with only a subtle increase in transcription of inflammation related genes in lungs and tracheal bronchial lymph nodes, while the level of expression of the majority of immune genes measured remained comparable to uninfected animals. Increasing viral load was observed in the blood and peripheral tissues at 16 and 21 DPI, respectively, indicating systemic viral dissemination. Clinical signs of MCF were observed between 28 and 35 DPI and the severity of lesions increased as disease progressed. Lesion scores were positively correlated with expression levels of ORF25, suggesting a contribution of viral replication in the pathogenesis of SA-MCF. Viral transcripts were observed in all tissues examined from 23 DPI to the end of the experiment at 35 DPI and expression levels of ORF25 were significantly higher in clinically infected animals as compared to pre-clinical stage. The data from this study provide a predictable viral-host interaction time course to test hypotheses concerning disease pathogenesis as well as mitigation of SA-MCF in susceptible species.

The anatomy of human teeth reflects its usage. Spatially resolved X-ray scattering permits quantitative studies of the characteristic arrangement of the anisotropic calcium phosphate crystallites and the collagen fibers within the hard tissues of the crown. The present study summarizes the distinctive nanometer-sized anatomical features of the tooth hard tissues including their interface taking advantage of spatially resolved synchrotron radiation-based small-angle X-ray scattering. The comparison of slices from eight teeth indicates a long-range organization of tooth nanostructures.

We have previously identified amelotin (AMTN) as a novel protein expressed predominantly during the late stages of dental enamel formation, but its role during amelogenesis remains to be determined. In this study we generated transgenic mice that produce AMTN under the amelogenin (Amel) gene promoter to study the effect of AMTN overexpression on enamel formation in vivo. The specific overexpression of AMTN in secretory stage ameloblasts was confirmed by Western blot and immunohistochemistry. The gross histological appearance of ameloblasts or supporting cellular structures as well as the expression of the enamel proteins amelogenin (AMEL) and ameloblastin (AMBN) was not altered by AMTN overexpression, suggesting that protein production, processing and secretion occurred normally in transgenic mice. The expression of Odontogenic, Ameloblast-Associated (ODAM) was slightly increased in secretory stage ameloblasts of transgenic animals. The enamel in AMTN-overexpressing mice was much thinner and displayed a highly irregular surface structure compared to wild type littermates. Teeth of transgenic animals underwent rapid attrition due to the brittleness of the enamel layer. The microstructure of enamel, normally a highly ordered arrangement of hydroxyapatite crystals, was completely disorganized. Tomes' process, the hallmark of secretory stage ameloblasts, did not form in transgenic mice. Collectively our data demonstrate that the overexpression of amelotin has a profound effect on enamel structure by disrupting the formation of Tomes' process and the orderly growth of enamel prisms.

Scrapie is the transmissible spongiform encephalopathy (TSE) of sheep and goats, and scrapie eradication in sheep is based in part on strong genetic resistance to classical scrapie. Goats may serve as a scrapie reservoir, and to date there has been no experimental inoculation confirming strong genetic resistance in goats. Two prion protein variants (amino acid substitutions S146 and K222) in goats have been significantly underrepresented in scrapie cases though present in scrapie-exposed flocks, and have demonstrated low cell-free protein conversion efficiency to the disease form (PrP(D)). To test degree of genetic resistance conferred in live animals with consistent exposure, we performed the first oral scrapie challenge of goats singly heterozygous for either PRNP S146 or K222. All N146-Q222 homozygotes became clinically scrapie positive by an average of 24months, but all S146 and K222 heterozygotes remain scrapie negative by both rectal biopsy and clinical signs at significantly longer incubation times (P<0.0001 for both comparisons). Recent reports indicate small numbers of S146 and K222 heterozygous goats have become naturally infected with scrapie, suggesting these alleles do not confer complete resistance in the heterozygous state but rather extend incubation. The oral challenge results presented here confirm extended incubation observed in a recent intracerebral challenge of K222 heterozygotes, and to our knowledge provide the first demonstration of extended incubation in S146 heterozygotes. These results suggest longer relevant trace-back histories in scrapie-eradication programs for animals bearing these alleles and strengthen the case for additional challenge experiments in both homozygotes to assess potential scrapie resistance.

Familial dilated cardiomyopathy is a primary myocardial disease that can result in the development of congestive heart failure and sudden cardiac death. Spontaneous animal models of familial dilated cardiomyopathy exist and the Doberman pinscher dog is one of the most commonly reported canine breeds. The objective of this study was to evaluate familial dilated cardiomyopathy in the Doberman pinscher dog using a genome-wide association study for a genetic alteration(s) associated with the development of this disease in this canine model. Genome-wide association analysis identified an area of statistical significance on canine chromosome 14 (p (raw) = 9.999e-05 corrected for genome-wide significance), fine-mapping of additional SNPs flanking this region localized a signal to 23,774,190-23,781,919 (p = 0.001) and DNA sequencing identified a 16-base pair deletion in the 5' donor splice site of intron 10 of the pyruvate dehydrogenase kinase 4 gene in affected dogs (p < 0.0001). Electron microscopy of myocardium from affected dogs demonstrated disorganization of the Z line, mild to moderate T tubule and sarcoplasmic reticulum dilation, marked pleomorphic mitochondrial alterations with megamitochondria, scattered mitochondria with whorling and vacuolization and mild aggregates of lipofuscin granules. In conclusion, we report the identification of a splice site deletion in the PDK4 gene that is associated with the development of familial dilated cardiomyopathy in the Doberman pinscher dog.

STATEMENT OF PROBLEM Porcelain fused to zirconia prostheses are widely used. However, porcelain chipping, spalling, fracture, and delamination are common clinical problems. Residual stresses of thermal origin have received attention, but clear data and firing guidelines remain absent. PURPOSE The purpose of this study was to measure the influence of heating and cooling protocols on the strength of porcelain fused to zirconia. MATERIAL AND METHODS A modified 4-point flexural testing technique was used to measure strength, and porcelain buttons were bonded to the beam between the 2 central loading points. Beams (n=54) were made of a tetragonal polycrystalline zirconium dioxide that was partially stabilized with an yttria core and a feldspathic dental porcelain. Three different heating rates and 3 different cooling regimens were used during firing. Two-way analysis of variance (ANOVA) was used to evaluate the 2 main effects of the heating and cooling regimens and their interaction with the delamination force (α=.05). The Tukey multiple comparisons test was used to identify differences among heating or cooling regimens. RESULTS During loading, the porcelain buttons separated from the zirconia beams because of delamination within the porcelain, which was close to the porcelain to zirconia interface. ANOVA revealed that the effects of the cooling regimen and heating rate had statistically significant effects on failure load (P<.05). The effect of the cooling regimen was greater than that of the heating regimen. CONCLUSIONS Slow cooling and slow heating regimens should be used when firing porcelain to zirconia. Cooling regimens were found to be more influential than heating rates. Failure was localized to the porcelain adjacent to the porcelain-zirconia interface, not to the interface itself, indicating that the residual stresses of thermal origin within the porcelain dominated. The preparation of zirconia with 50 μm aluminum oxide at a pressure of 0.34 MPa was sufficient to prevent interfacial failure.

Visna/Maedi, or ovine progressive pneumonia (OPP) as it is known in the United States, is an incurable slow-acting disease of sheep caused by persistent lentivirus infection. This disease affects multiple tissues, including those of the respiratory and central nervous systems. Our aim was to identify ovine genetic risk factors for lentivirus infection. Sixty-nine matched pairs of infected cases and uninfected controls were identified among 736 naturally exposed sheep older than five years of age. These pairs were used in a genome-wide association study with 50,614 markers. A single SNP was identified in the ovine transmembrane protein (TMEM154) that exceeded genome-wide significance (unadjusted p-value 3×10(-9)). Sanger sequencing of the ovine TMEM154 coding region identified six missense and two frameshift deletion mutations in the predicted signal peptide and extracellular domain. Two TMEM154 haplotypes encoding glutamate (E) at position 35 were associated with infection while a third haplotype with lysine (K) at position 35 was not. Haplotypes encoding full-length E35 isoforms were analyzed together as genetic risk factors in a multi-breed, matched case-control design, with 61 pairs of 4-year-old ewes. The odds of infection for ewes with one copy of a full-length TMEM154 E35 allele were 28 times greater than the odds for those without (p-value<0.0001, 95% CI 5-1,100). In a combined analysis of nine cohorts with 2,705 sheep from Nebraska, Idaho, and Iowa, the relative risk of infection was 2.85 times greater for sheep with a full-length TMEM154 E35 allele (p-value<0.0001, 95% CI 2.36-3.43). Although rare, some sheep were homozygous for TMEM154 deletion mutations and remained uninfected despite a lifetime of significant exposure. Together, these findings indicate that TMEM154 may play a central role in ovine lentivirus infection and removing sheep with the most susceptible genotypes may help eradicate OPP and protect flocks from reinfection.

Purpose: Fracture is a frequent complication of resinous prostheses. The purpose of this study was to evaluate the effect of thickness on flexural strength of a resinous prosthesis containing a prosthetic tooth. Materials and Methods: Beam-shaped specimens 65-mm long, 12-mm wide, and 1, 2, 3, 4, or 6 mm in thickness were made from high-impact strength polymethyl methacrylate denture base material, each containing a resin-based molar prosthetic tooth at the center of the beam. A group of 3-mm-thick specimens without a prosthetic tooth (n = 7) were also made. Specimens were aged artificially, loaded in three-point flexure, examined fractographically, and analyzed. Results: The 1- and 2-mm-thick beams underwent considerable deformation at low loads. Maximum loads varied considerably from 0.6 kg (1-mm beams) to 38 kg (6-mm beams). The 3-, 4-, and 6-mm beam groups all underwent brittle fracture, with mean relative flexural strengths of approximately 73 MPa. Denture teeth reduced the relative flexural strength of resin beams by 0.7x. Fracture initiation sites were generally at tiny surface defects, but did not directly involve denture teeth. Denture resin fracture toughness was 3.2 MPa m1/2, and modulus of rupture was 104 MPa. Conclusion: Denture teeth substantially decreased the strength of resinous beams. Increased thickness markedly increased the load-bearing capacity of resinous beams containing denture teeth. Beams less than 2 mm in thickness with denture teeth were weakened substantially more than comparable beams of 2 mm or more in thickness. Surface finish was of critical importance. Fracture toughness was calculated fractographically, facilitating future forensic examination of clinically failed resinous prostheses. Int J Prosthodont 2012;25:53-59.

Flow coefficients v_{n} for n=2, 3, 4, characterizing the anisotropic collective flow in Au+Au collisions at sqrt[s_{NN}]=200  GeV, are measured relative to event planes Ψ_{n}, determined at large rapidity. We report v_{n} as a function of transverse momentum and collision centrality, and study the correlations among the event planes of different order n. The v_{n} are well described by hydrodynamic models which employ a Glauber Monte Carlo initial state geometry with fluctuations, providing additional constraining power on the interplay between initial conditions and the effects of viscosity as the system evolves. This new constraint can serve to improve the precision of the extracted shear viscosity to entropy density ratio η/s.

The equid hemoprotozoan parasite Theileria equi is endemic in most regions worldwide. Infection of horses is a cause of significant economic loss due to costs associated with disease and restriction of trade with non-endemic nations. The ability of certain drugs such as imidocarb dipropionate to eliminate persistent T. equi infection and transmission risk is controversial. The anti-protozoal agent ponazuril has been used successfully to treat equine Sarcosystis neurona and Toxoplasma gondii. The hypothesis that ponazuril inhibits replication of T. equi in vitro was tested. T. equi infected equine erythrocyte cultures were treated with ponazuril at multiple concentrations. Cessation of parasite replication was observed over a 5-day period and the degree of inhibition was variable between drug concentrations. Ponazuril inhibited T. equi in erythrocyte culture at all concentrations tested but parasite elimination required at least 500μg/mL. The high dose of ponazuril required for in vitro inhibition likely limits its ability to control or clear T. equi infection in vivo, however additional research to evaluate related drugs is warranted.