Genome-wide association analysis for susceptibility to infection by Mycobacterium avium ssp. paratuberculosis in US Holsteins.

Paratuberculosis, or Johne's disease, is a chronic, granulomatous, gastrointestinal tract disease of cattle and other ruminants caused by the bacterium Mycobacterium avium subspecies paratuberculosis (MAP). Control of Johne's disease is based on programs of testing and culling animals positive for infection with MAP and concurrently modifying management to reduce the likelihood of infection. The current study was motivated by the hypothesis that genetic variation in host susceptibility to MAP infection can be dissected and quantifiable associations with genetic markers identified. Two separate GWAS analyses were conducted, the first using 897 genotyped Holstein artificial insemination sires with phenotypes derived from incidence of MAP infection among daughters based on milk ELISA testing records. The second GWAS analysis was a case-control design using US Holstein cows phenotyped for MAP infection by serum ELISA or fecal culture tests. Cases included cows positive for either serum ELISA, fecal culture, or both. Controls consisted of animals negative for all tests conducted. A total of 376 samples (70 cases and 306 controls) from a University of Minnesota Johne's management demonstration project and 184 samples (76 cases and 108 controls) from a Michigan State University study were used. Medium-density (sires) and high-density (cows) genotype data were imputed to full genome sequence for the analyses. Marker-trait associations were analyzed using the single-step (ss)GWAS procedure implemented in the BLUPF90 suite of programs. Evidence of significant genomic contributions for susceptibility to MAP infection were observed on multiple chromosomes. Results were combined across studies in a meta-analysis, and increased support for genomic regions on BTA7 and BTA21 were observed. Gene set enrichment analysis suggested pathways for antigen processing and presentation, antimicrobial peptides and natural killer cell-mediated cytotoxicity are relevant to variation in host susceptibility to MAP infection, among others. Genomic prediction was evaluated using a 5-fold cross-validation, and moderate correlations were observed between genomic breeding value predictions and daughter averages (∼0.43 to 0.53) for MAP infection in testing data sets. These results suggest that genomic selection against susceptibility to MAP infection is feasible in Holstein cattle.

The role of subchondral bone, and its histomorphology, on the dynamic viscoelasticity of cartilage, bone and osteochondral cores.

OBJECTIVE: Viscoelastic properties of articular cartilage have been characterised at physiological frequencies. However, studies investigating the interaction between cartilage and subchondral bone and the influence of underlying bone histomorphometry on the viscoelasticity of cartilage are lacking. METHOD: Dynamic Mechanical Analysis (DMA) has been used to quantify the dynamic viscoelasticity of bovine tibial plateau osteochondral cores, over a frequency sweep from 1 to 88 Hz. Specimens (approximately aged between 18 and 30 months) were neither osteoarthritic nor otherwise compromised. A maximum nominal stress of 1.7 MPa was induced. Viscoelastic properties of cores have been compared with that of its components (cartilage and bone) in terms of the elastic and viscous components of both structural stiffness and material modulus. Micro-computed tomography scans were used to quantify the histomorphological properties of the subchondral bone. RESULTS: Opposing frequency-dependent loss stiffness, and modulus, trends were witnessed for osteochondral tissues: for cartilage it increased logarithmically (P < 0.05); for bone it decreased logarithmically (P < 0.05). The storage stiffness of osteochondral cores was logarithmically frequency-dependent (P < 0.05), however, the loss stiffness was typically frequency-independent (P > 0.05). A linear relationship between the subchondral bone plate (SBP) thickness and cartilage thickness (P < 0.001) was identified. Cartilage loss modulus was linearly correlated to bone mineral density (BMD) (P < 0.05) and bone volume (P < 0.05). CONCLUSION: The relationship between the subchondral bone histomorphometry and cartilage viscoelasticity (namely loss modulus) and thickness, have implications for the initiation and progression of osteoarthritis (OA) through an altered ability of cartilage to dissipate energy.

Structured three-dimensional co-culture of mesenchymal stem cells with chondrocytes promotes chondrogenic differentiation without hypertrophy.

OBJECTIVE: This study investigated a novel approach to induce chondrogenic differentiation of human mesenchymal stem cells (hMSC). We hypothesized that a structured three-dimensional co-culture using hMSC and chondrocytes would provide chondroinductive cues to hMSC without inducing hypertrophy. METHOD: In an effort to promote optimal chondrogenic differentiation of hMSC, we created bilaminar cell pellets (BCPs), which consist of a spherical population of hMSC encased within a layer of juvenile chondrocytes (JC). In addition to histologic analyses, we examined proteoglycan content and expression of chondrogenic and hypertrophic genes in BCPs, JC pellets, and hMSC pellets grown in the presence or absence of transforming growth factor-ß (TGFß) following 21 days of culture in either growth or chondrogenic media. RESULTS: In either growth or chondrogenic media, we observed that BCPs and JC pellets produced more proteoglycan than hMSC pellets treated with TGFß. BCPs and JC pellets also exhibited higher expression of the chondrogenic genes Sox9, aggrecan, and collagen 2A1, and lower expression of the hypertrophic genes matrix metalloproteinase-13, Runx2, collagen 1A1, and collagen 10A1 than hMSC pellets. Histologic analyses suggest that JC promote chondrogenic differentiation of cells in BCPs without hypertrophy. Furthermore, when cultured in hypoxic and inflammatory conditions intended to mimic the injured joint microenvironment, BCPs produced significantly more proteoglycan than either JC pellets or hMSC pellets. CONCLUSION: The BCP co-culture promotes a chondrogenic phenotype without hypertrophy and, relative to pellet cultures of hMSCs or JCs alone, is more resistant to the adverse conditions anticipated at the site of articular cartilage repair.

Debonding force and deformation of two multi-stranded lingual retainer wires bonded to incisor enamel: an in vitro study.

The aim of the study was to examine, in vitro, the effect of a vertical force on the debond force and deformation of two multi-stranded wires bonded to the lingual enamel of lower incisor teeth. Two different stainless steel wires were used, 0.016 × 0.022 inch (Bond-A-Braid® Reliance Orthodontic Products) and a three-stranded 0.0175 inch wire (Ortho Technology). An in vitro model was used to simulate a vertical force at the interdental wire. Twenty-six pairs of incisors were placed in two groups. A 15 mm length of wire was bonded to the lingual surfaces of each pair of incisors using a common bonding technique. A vertical force was applied to the midpoint of the interdental wire, using an Instron universal testing machine. The failure characteristics examined included the maximum force for debond, the degree of wire deformation, and the site of failure. Significance was predetermined at P < 0.05 and multiple comparisons indicated no significant differences (P = 0.147) in the initial mean bond strength between the 0.0175 inch (41.44 N) and 0.016 × 0.022 inch (37.70 N) wires. The main failure type for both the initial and second debond events was fracture of composite bond at the wire-composite interface, cohesive failure. Both wires exhibited similar mean degrees of deflection of 1.30 and 1.51 mm for the 0.0175 inch and 0.016 × 0.022 inch wires, respectively. Rebonding to enamel resulted in significantly lower (P = 0.001) mean bond strength for both wires, 0.0175 inch (13.86 N) and 0.016 × 0.022 inch (14.17 N) in comparison with the initial bond strength. Rebonding to previously bonded enamel may be unpredictable and may lead to higher failure rates of bonded lingual retainers.

Maxillary premolar resorption by canines: three case reports.

Three unusual cases of maxillary premolar root resorption are reported. Three teenage patients were referred to the orthodontic department for management of ectopic maxillary canines. Radiographic examination revealed unilateral premolar root resorption in all three patients. This represents an unusual finding. Whereas the prevalence of maxillary lateral incisor root resorption secondary to palatally ectopic canines has been reported, the prevalence of premolar root resorption is unknown. This report discusses the findings in the context of the available literature. The postulated aetiology and the need for early diagnosis are highlighted.

Stabilisation of the unstable fractured zygomatic arch with a Kirschner wire.

Although relatively uncommon, isolated fractures of the zygomatic arch can sometimes be difficult to stabilise following reduction. We present a simple method of stabilisation using a Kirschner wire.

Schinzel-Giedion syndrome: interesting facial and orodental features, and dental management.

Schinzel-Giedion syndrome comprises multiple congenital anomalies. The orofacial features include coarse facies, frontal bossing, ocular hypertelorism, anterior open bite and macrodontia. Two cases are presented in which the presence of specific craniofacial anomalies with bilateral hydronephrosis confirmed the diagnosis. In one patient, bottle-feeding was associated with caries in maxillary central and lateral incisors, but the second patient was permanently tube fed and did not experience any dental caries. Clinical management of these patients requires a coordinated approach from a team of medical and dental specialists.

Modulation of cell interactions with extracellular matrix by lysophosphatidic acid and sphingosine 1-phosphate.

Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (SPP) are lipid mediators released upon platelet activation. The concentration of LPA in serum is estimated at 1-10 microM whereas the concentration in plasma is considerably less. The SPP concentration in serum is 0.5 microM, approximately two-fold higher than the plasma concentration. The lipids are present during tissue injury and promote cellular processes involved in wound repair. LPA and SPP have multiple effects on cells, many of which are pertinent to wound healing and require that the cells interact in some fashion with components of the extracellular matrix. This review focuses on modulation of cell adhesion, cell migration, collagen gel contraction, and fibronectin matrix assembly by LPA and SPP.

Contraction of collagen matrices mediated by alpha2beta1A and alpha(v)beta3 integrins.

The (beta)1-null fibroblastic cell line GD25 and its derivatives were studied to gain an understanding of the roles of (beta)1 and (beta)3 integrins in the initial (1-hour) contraction of collagen gels. Stable transfectants of GD25 cells expressing the (beta)1A splice variant of (beta)1 ((beta)1A-GD25) did not express (alpha)2(beta)1A and did not adhere to collagen. After transfection of (alpha)2 into (beta)1A-GD25 cells, the (alpha)2(beta)1A-GD25 transfectants contracted collagen gels in the presence of serum, whereas (beta)1A-GD25 cells did not. The GD25 parental cells, however, also contracted collagen gels. Collagen gel contraction by GD25 cells was blocked by antibodies to (alpha)v(beta)3 or a RGD-containing peptide, indicating that (alpha)v(beta)3 is the integrin responsible for mediation of contraction by GD25 cells. Collagen gel contraction by (alpha)2(beta)1A-GD25 cells was not inhibited by antibodies to (alpha)v(beta)3 or RGD-containing peptide, but was inhibited by anti-(alpha)2 antibody. Flow cytometry demonstrated negligible expression of (alpha)v(beta)3 by (beta)1A-GD25 and (alpha)2(beta)1A-GD25 cells when compared to GD25 cells. Platelet derived growth factor (PDGF) and sphingosine-1-phosphate (S1P) enabled gel contraction by (alpha)2(beta)1A-GD25 and GD25 cells, respectively, in the absence of serum. PDGF-stimulated contraction by (alpha)2(beta)1A-GD25 cells was attenuated in the presence of inhibitors of phosphatidylinositol-3-kinase whereas such inhibitors had no effect on S1P-stimulated contraction by GD25 cells. These experiments using the (beta)1-null GD25 cells and (beta)1A and (alpha)2(beta)1A transfectants demonstrate that (alpha)2(beta)1A and (alpha)v(beta)3 independently mediate collagen gel contraction and are regulated by different serum factors and signaling pathways.

Cervical interbody fusion cages. An animal model with and without bone morphogenetic protein.

STUDY DESIGN: The Alpine goat model for multilevel anterior cervical discectomy and fusion was used to analyze the use of an intervertebral fusion device to promote an arthrodesis after anterior cervical discectomy. Comparisons were drawn with biomechanical, histologic, and radiographic data. OBJECTIVES: To analyze the use of an intervertebral fusion device, with and without a bone graft substitute, to promote an arthrodesis anterior cervical discectomy. SUMMARY OF BACKGROUND DATA: In previous studies, the goat cervical spine has proven to be an excellent model for examining the healing of fusions using bone grafts, instrumentation, or bone substitutes. METHODS: Three-level anterior cervical dissectomies were performed on 21 mature Alpine goats. Three treatment groups of seven goats each were used. Group I used a standard titanium cervical BAK device filled with autogenous bone graft. Group II used a hydroxyapatite-coated BAK device filled with autogenous bone graft. Group III used a BAK device filled with recombinant human bone morphogenetic protein-2. RESULTS: Radiographically, no cages became displaced. Lucencies were seen around 3 of the 21 cages in Group 1, 4 cages in Group II, and none in Group III. Fluorochrome analysis revealed that the recombinant human bone morphogenetic protein-2-filled cages had an accelerated rate of bone growth around and through each cage-vertebral body interface at 3 weeks. A successful arthrodesis was also more likely with a recombinant human bone morphogenetic protein-2-filled cage (95%) than the hydroxyapatite-coated (62%) or the standard (48%) cage. Biomechanical stiffness testing did not reveal any statistically significant differences between the three groups. There was a tendency for successfully arthrodesed interspaces to be stiffer than those that were not. CONCLUSIONS: The use of a threaded intervertebral fusion cage, with or without hydroxyapatite coating, filled with autogenous bone graft provides a fusion rate that is slightly better than those previously reported using autogenous interbody bone grafts with or without plate stabilization. Recombinant human bone morphogenetic protein-2-filled cages resulted in a much higher arthrodesis rate and accelerated bone formation compared with either autogenous bone-filled BAK devices, or autogenous interbody bone grafts with or without plate stabilization.