Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity

OBJECTIVES: To characterize equine bone marrow (BM)-derived mesenchymal stem cell (MSC) growth characteristics and frequency as well as their adipogenic and osteogenic differentiation potential. STUDY DESIGN: In vitro experimental study. ANIMALS: Foals (n=3, age range, 17-51 days) and young horses (n=5, age range, 9 months to 5 years). METHODS: Equine MSCs were harvested and isolated from sternal BM aspirates and grown up to passage 10 to determine cell-doubling (CD) characteristics. Limit dilution assays were performed on primary and passaged MSCs to determine the frequency of colony-forming units with a fibroblastic phenotype (CFU-F), and the frequency of MSC differentiation into adipocytes (CFU-Ad) and osteoblasts (CFU-Ob). RESULTS: Initial MSC isolates had a lag phase with a significantly longer CD time (DT=4.9+/-1.6 days) compared with the average DT (1.4+/-0.22 days) of subsequent MSC passages. Approximately 1 in 4224+/-3265 of the total nucleated BM cells displayed fibroblast colony-forming activity. Primary MSCs differentiated in response to adipogenic and osteogenic inductive conditions and maintained their differentiation potential during subsequent passages. CONCLUSIONS: The frequency, in vitro growth rate, and adipogenic and osteogenic differentiation potential of foals and young adult horses are similar to those documented for BM MSCs of other mammalian species. CLINICAL RELEVANCE: The results have direct relevance to the use of BM as a potential source of adult stem cells for tissue engineering applications in equine veterinary medicine.     

Differences in metabolic parameters and gene expression related to osteochondrosis/osteoarthrosis in pigs fed 25-hydroxyvitamin D3

Osteochondrosis/osteoarthrosis (OC/OA) are common terms for various joint pathologies that occur in pigs. Pathologies that may contribute to these disorders have been described, but the primary cause(s) remain unknown. We hypothesised that as OC has some similarities to dyschondroplasia, which involves a failure of growth plate chondrocytes to fully differentiate and hypertrophy, treatment with 25-hydroxyvitamin D3 (25-D) might reduce the incidence and/or severity of lesions in pigs, as it does in chickens with dyschondroplasia. Control pigs were fed a commercial diet ad libitum. In the treated group this diet was supplemented with 25-D at 0.1 mg/kg. Ten pigs from each of the control and treated groups were sampled at 7, 12, 16 and 21 weeks. Treatment with 25-D had no effect on the incidence or severity of OC/OA lesions. Cartilage dry weight, total collagen content and proteoglycan content, and plasma levels oftotal calcium, inorganic phosphorous, vitamin C, insuline-like growth factor-I, parathyroid hormone and tumour necrosis factor alpha were unaffected by treatment. In addition, none of these parameters were correlated with the incidence or severity of OC/OA lesions. The mRNA expression levels of 21 out of 23 genes assayed by RT-PCR were unaltered in articular cartilage from OA lesion samples as compared to normal articular cartilage. However, collagen type II was reduced and collagen type X increased in OA lesion and near lesion samples. These results suggest that OA in pigs may share some features of osteoarthritis in other mammalian species.     

QUANTITATIVE ASSESSMENT OF SURGICALLY INDUCED MITRAL REGURGITATION USING RADIONUCLIDE VENTRICULOGRAPHY AND FIRST PASS RADIONUCLIDE ANGIOGRAPHY

Radionuclide ventriculography has been used in humans to evaluate valvular incompetency. The stroke volume ratio, derived from the radionuclide ventriculogram, is used to quantify the severity of mitral regurgitation (MR). Previous studies conducted in humans have shown that left to right stroke volume ratio increases as the severity of MR increases. In this study, we evaluated radionuclide ventriculography as a noninvasive method to detect MR in dogs with surgically created mitral insufficiency. Six male and three female adult, conditioned mongrel dogs were used. Scintigraphic studies were performed prior to and 4 weeks after surgically created MR. Because of the overlap of the left and right ventricles when viewed from a left lateral position, we combined data from a first-pass radionuclide angiocardiogram with the radionuclide ventriculogram to obtain a corrected stroke volume ratio. Blood flow transit parameters were also derived from the first-pass radionuclide angiocardiogram. Standard left ventricular functional indices were also measured from the radionuclide ventriculogram. On the left lateral view of the heart, 25 to 30% of the right ventricular volume overlaps the left ventricle. After correcting for the overlap, the stroke volume ratio of normal dogs was 1.17±0.178 (mean±SD), which increased to 2.06±0.41 (mean±SD) (p<.001) 4 weeks after creation of MR. The was no significant change in left ventricular ejection fraction or peak rate of ejection following MR. The transit times of blood through the left ventricle were measured from the first-pass radionuclide angiocardiogram and were expressed as half-time clearance, peak clearance rate, and time to peak clearance rate. The baseline half-time clearance was 2.07±0.71 s (mean±SD), which increased to 6.70±4.89 s (mean±SD) (p=.02) after creation of MR. The baseline peak clearance rate was 49.75±8.96 cts/s (mean±SD), which decreased to 23.12±6.84 cts/s (mean±SD) (p<.001) after creation of MR. Stroke volume ratios significantly increased following creation of MR. Blood flow transit through the left ventricle slowed following creation of MR. The variability of these parameters were small in the baseline studies, suggesting these techniques may be clinically useful to gauge the severity of MR in dogs.     

Spontaneous neoplasms of lagomorphs.

Many of the same neoplastic processes reported in companion animal medicine have been documented in lagomorphs. With only a few reports in the literature regarding rabbits and hares, the behavior of many of these tumors cannot be predicted. Further, based on limited current knowledge, the practitioner is left to make decisions for diagnostics and therapeutics in these cases based upon current recommendations for companion animals with adjustments for lagomorph physiology. We hope that this review of lagomorph neoplasia will inform the general practitioner of the occurrence of these tumors, promote further documentation and research of these processes in rabbits, and encourage the practitioner of rabbit medicine along with rabbit owners to consult with veterinary oncologists regarding neoplastic processes in rabbits rather than just giving a guarded prognosis.     

Measurement of angles of abduction for diagnosis of shoulder instability in dogs using goniometry and digital image analysis

OBJECTIVE: To compare abduction angles of shoulders with medial instability and unaffected shoulders in the same dogs and in age- and breed-matched dogs. STUDY DESIGN: Case-control study. ANIMALS: Dogs with medial instability of the shoulder (n=33) and 26 control dogs. METHODS: Dogs were sedated and positioned in lateral recumbency with both scapulas parallel to the table. With the elbow and shoulder in extension, the non-recumbent limb was maximally abducted and the angle between the scapular spine and lateral aspect of the brachium measured with a goniometer; a digital image was taken from the cranial aspect. Both techniques were performed in triplicate by 2 examiners. Mean abduction angles for each shoulder were determined from goniometric measurements and image analysis. Data were analyzed for significant differences between affected and unaffected shoulders, measurement techniques, and examiners. Strength of correlation between measurement techniques was determined. RESULTS: Mean abduction angles for shoulders with instability (53.7+/-4.7 degrees goniometric, 51.2+/-4.9 degrees image) were significantly (P<.001) larger than for all unaffected shoulders (32.6+/-2.0 degrees goniometric, 30.9+/-2.3 degrees image). In dogs diagnosed with instability, affected shoulders had significantly (P<.001) larger abduction angles than the contralateral (unaffected) shoulders. No significant differences were identified between right and left shoulders for control dogs, measurement techniques, or examiners. A strong (r=0.90) significant (P<.001) positive correlation between measurement techniques was noted. CONCLUSIONS: Shoulder abduction angles measured under sedation provide objective data for diagnosis of shoulder instability in dogs. Shoulders with clinical and arthroscopic evidence of medial instability have significantly higher abduction angles than shoulders that are considered normal. CLINICAL RELEVANCE: Determination of shoulder abduction angles should be included in the diagnostic protocol for forelimb lameness assessment in dogs.     

Pesticide confirmation by triple stage quadrupole mass spectrometry: etrimfos and dimethoate

The production of a protonated molecular ion, MH+, for a pesticide under investigation is often preferred for primary identification purposes. However, the lack of fragment ions under such chemical ionization conditions can place the burden of proof on a single ion species for confirmation. Although a single ion representing the molecule at the correct retention time on a packed column or high-resolution capillary column might seem to have furnished sufficient evidence for unambiguous confirmation, in a number of cases additional specificity can improve the analytical result. With the introduction into commerce of triple stage quadrupole (TSQ) instruments, the possibility of improving the degree of specificity by chemical ionization has emerged for practical pesticide residue confirmation analysis. Use of collision-activated dissociation (CAD) experiments to form daughter ions from the protonated molecular ion of 2 representative organophosphorus pesticides has provided a second dimensional plane of characterization for confirmatory purposes at incurred residue levels.     

B-cell lymphoma in a dog with ehrlichiosis (Ehrlichia canis) and systemic histoplasmosis (Histoplasma capsulatum)

A mixed breed dog treated for ehrlichiosis and systemic histoplasmosis developed a refractory thrombocytopenia. When an abdominal mass was detected, exploratory laparotomy and biopsies confirmed lymphoma, which on immunohistochemical stains was determined to be of B-cell origin. Conceivably, the B-cell lymphoma in this dog was associated with chronic inflammation from ehrlichiosis, histoplasmosis, or both.     

Impact of diet on development of bronchial-associated immunity in the neonatal piglet

Bronchial-associated immune development is critically important to protect neonates from respiratory infections. Herein, bronchial-associated immune development in formula-fed and sow-reared pigs is described. Colostrum-fed newborn piglets were fed medicated sow milk replacer formula beginning at 48 h of life or remained with the sow. Blood and tissues were sampled at one-week (d7) and three-weeks (d21) of age. Lymphocyte subpopulations, including T helper 2, cytotoxic T, memory T, and NK cells, in peripheral blood, mediastinal lymph nodes, and thoracic lymph nodes were identified using flow cytometry. Additionally, IL-1β, IL-6, IL-12, TNFα, TGF-β1, TGF-β2, IFNα, IFNβ, and dectin gene expression were analyzed by quantitative RT-PCR. Total IgG, IgM, and IgA concentrations in serum were analyzed. Dietary and developmental effects were observed. This set of baseline measurements provides a framework for future respiratory challenge studies where the effects of diet on the neonate's ability to resist and/or recover from infection can be tested.