40 research outputs found

    Surface Glycan Pattern of Canine, Equine, and Ovine Bone Marrow-Derived Mesenchymal Stem Cells

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    The use of bone marrow-derived mesenchymal stem cells (MSCs) for clinical and experimental studies is increasing, but full characterization of MSCs in veterinary species is hindered by the variability in species-specific cell surface marker expression and antibody cross reactivity. Recent studies demonstrated that the glycans in the glycocalyx of MSCs are promising candidates as cell biomarkers. In the present study, we analysed the glycocalyx of canine MSCs (cMSCs), ovine MSCs (oMSCs), and equine MSCs (eMSCs) by using a cell microarray procedure in which MSCs were spotted on microarray slides and incubated with a panel of 14 biotinylated lectins and Cy3-conjugated streptavidin. The signal intensity was then detected by using a microarray scanner. The lectin-binding signals indicated that the MSC surface of the investigated species contained both N- and O-linked glycan types, with N-glycosylation predominating over O-glycosylation and fucosylation being more abundant than sialylation. Relative quantification revealed an interspecific difference between these glycans. In addition, cMSCs expressed more α2,3-linked sialic acid (MAL II), terminal lactosamine (RCA120), and α1,6 and α1,3 fucosylated oligosaccharides (PSA, LTA); oMSCs exhibited more T antigen (Jacalin), GalNAcα1,3(LFucα1,2)Galβ1,3/4GlcNAcβ1 (DBA), chitotriose (succinylated WGA), and α1,2-linked fucose (UEA I); and eMSCs showed a higher density of α2,6 sialic acids (SNA) and high mannose N-glycans (Con A). By using cell microarray methodology, we have for the first time demonstrated differences in the glycosylation profiles of cMSC, oMSC, and eMSC surfaces. These results could be valuable as resources and references for MSC differentiation and molecular remodeling in clinical cell-based therapy and tissue engineering studies

    Treatment of Monolateral Legg-Calvé-Perthes Disease with Autologous Bone Marrow Mononuclear Cells in 32 Dogs

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    Objective In the present study, we report our results of the use of autologous bone marrow mononuclear cells (BMMCs) as a minimally invasive treatment for Legg-Calvé-Perthes in dogs. Study Design In accordance with Ljunggren's scale, inclusion criteria were determined by clinical condition and radiographic features of the disease, resulting in 32 dogs enrolled in this retrospective study from 2007 to 2019. Bone marrow was collected from each dog from the iliac crest and the mononuclear fraction was separated with density gradient centrifugation. The mean number of BMMCs was 104.7 ± 46.5 × 106 cells. The mean BMMC colony-forming units were 71.6 ± 51.9 × 102/mL. Cells were suspended in fibrin glue before BMMC administration and implanted via transcutaneous injection under computed tomography or radiographic guidance, using a Jamshidi needle inserted through the femoral head and neck. Results A progressive reduction of pain within 3 weeks after BMMC administration was observed in 28 patients, with gradually increased weight bearing on the affected limb. In four dogs, however, pain and lameness persisted and at 3 months post-BMMC injection, femoral head and neck resection was performed. Histological and immunohistochemical studies were done on samples from those four dogs, which showed evidence of formation of new cartilage and subchondral bone in the area where cells were implanted. Clinical Significance Based on these results, BMMC therapy may be considered as effective and minimally invasive treatment option for LCPD in dogs

    Role of Tibial Tuberosity Fracture/Fissure through the Maquet Hole in Stifle Osteoarthritis after Porous Tibial Tuberosity Advancement in Dogs at Mid-Term Follow-Up

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    Tibial tuberosity advancement (TTA) is used to treat cranial cruciate ligament rupture of the stifle joint in dogs. Tibial tuberosity fracture/fissure is a complication of TTA that may have a favorable prognosis. The aim of this study was to detect how tibial tuberosity fracture/fissure through the Maquet hole worsens the progression of osteoarthritis (OA) in the stifle joint of dogs treated with porous TTA. Seventeen cases were included in the study, divided into two groups. The first group (n = 10) included subjects that had tibial tuberosity fracture/fissure through the Maquet, and the second group included subjects that had no complications (n = 7). Both groups showed significant progression compared to OA at 3 months after surgery. We observed that at T0, the control group showed a higher level of OA. For this reason, we normalized the OA scores, evaluating the percentage difference from T0 and T1. We verified that there were no statistically significant differences between the two groups. The results confirm that OA progression in subjects undergoing TTA was not significantly influenced by fracture/fissure of the tibial tuberosity through the Maquet hole. Therefore, fracture fissure through the Maquet hole should be considered as a common minor complication during TTA

    Effects of a probiotic on the morphology and mucin composition of pig intestine

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    Although the use of probiotics in human and animal medicine is growing, their mode of action remains poorly understood. This study examined the effects of a multi-strain probiotic (SLAB51TM) on the morphology and carbohydrate composition of mucins secreted by goblet cells of intestinal crypts in growingfinishing pigs. Sections of duodenum, caecum and colon from pigs fed for 12 weeks with an orally administered control basal diet (No-Pro) or one with a probiotic blend (Pro) were processed for microscopic analysis and stained with (1) haematoxylin-eosin for structural and morphometrical investigation; (2) conventional histochemistry (periodic acid-Schiff, Alcian Blue pH 2.5, high iron diamine staining) for neutral, acidic nonsulphated, and sulphated mucin analysis; and (3) FITClabelled MAA-II and SNA lectins for α2,3- and α2,6-sialomucin identification. Compared with No-Pro samples, Pro samples displayed (1) increased goblet cell numbers in all investigated tract crypts; (2) an increase in acidic non-sulphomucins but a decrease in neutral, sulphated and α2,6-sialomucin-secreting goblet cells in the duodenum; (3) decreased crypt depth, an increase in α2,6-sialomucin secretory goblet cells, and a loss of goblet cell-secreting α2,3-sialomucins, which appeared on the apical surface of crypt fundus epithelial cells in the caecum; and (4) an increase in α2,6- sialomucinproducing goblet cells in the colon. Results suggest that treatment with SLAB51TM induces region-specific changes in the morphology and carbohydrate composition of mucins secreted along intestinal tracts of growing-finishing pigs. These changes could ameliorate the health status of the animals, which displayed higher growth performance and meat quality than controls (Tufarelli et al., 2017).Although the use of probiotics in human and animal medicine is growing, their mode of action remains poorly understood. This study examined the effects of a multi-strain probiotic (SLAB51TM) on the morphology and carbohydrate composition of mucins secreted by goblet cells of intestinal crypts in growingfinishing pigs. Sections of duodenum, caecum and colon from pigs fed for 12 weeks with an orally administered control basal diet (No-Pro) or one with a probiotic blend (Pro) were processed for microscopic analysis and stained with (1) haematoxylin-eosin for structural and morphometrical investigation; (2) conventional histochemistry (periodic acid-Schiff, Alcian Blue pH 2.5, high iron diamine staining) for neutral, acidic nonsulphated, and sulphated mucin analysis; and (3) FITClabelled MAA-II and SNA lectins for α2,3- and α2,6- sialomucin identification. Compared with No-Pro samples, Pro samples displayed (1) increased goblet cell numbers in all investigated tract crypts; (2) an increase in acidic non-sulphomucins but a decrease in neutral, sulphated and α2,6-sialomucin-secreting goblet cells in the duodenum; (3) decreased crypt depth, an increase in α2,6-sialomucin secretory goblet cells, and a loss of goblet cell-secreting α2,3-sialomucins, which appeared on the apical surface of crypt fundus epithelial cells in the caecum; and (4) an increase in α2,6-sialomucinproducing goblet cells in the colon. Results suggest that treatment with SLAB51TM induces region-specific changes in the morphology and carbohydrate composition of mucins secreted along intestinal tracts of growing-finishing pigs. These changes could ameliorate the health status of the animals, which displayed higher growth performance and meat quality than controls (Tufarelli et al., 2017)

    Probiotic supplementation affects the glycan composition of mucins secreted by Brunner's glands of the pig duodenum.

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    The effect of a dietary probiotic blend on the carbohydrate composition of mucins secreted by the Brunner's glands in the duodenum of growing-finishing pigs was investigated by means of conventional (periodic acid-Schiff, Alcian Blue pH 2.5, high iron diamine staining) and lectin (15 lectins) histochemistry. Pigs were assigned to two dietary treatments: a control basal diet without the probiotic blend (No-Pro) and a test diet that included the probiotic blend (Pro). Duodenal tissue fragments were fixed in 4% phosphate-buffered-saline-buffered paraformaldehyde, dehydrated through a graded alcohol series, and embedded in paraffin wax. The secretory cells of the Brunner's glands from No-Pro pigs primarily produced neutral glycoproteins and a small amount of acidic non-sulphated mucins. This glycan pattern was opposite that of the Brunner's glands from Pro animals. A comparison of lectin-binding profiles of the secretory cells of Brunner's glands in these two groups showed that in Pro pigs, there was (i) a decrease in N-linked glycans containing α1,2-linked fucose (Con A, UEA I); (ii) a loss of complex types of N-glycans (PHA-L, PHA-E) terminating with lactosamine (RCA120), α1,6- and α1,3-linked fucose (LTA), and α-galactose (GSA I-B4), as well as of O-glycans with terminal Galβ1,3GalNAc (PNA); and (iii) an increase in O-glycans containing GalNAc HPA. No-Pro and Pro samples showed no change in the expression of α2,6 sialoglycans and terminal GlcNAc residues and no affinity for MAL II, DBA, and SBA. These results indicate that probiotic supplementation affects the glycan composition of mucins produced in the Brunner's glands of growing-finishing pigs. These changes could effectively act on the gastrointestinal function and health status of these animals because the probiotic blend induced higher growth performance and meat quality in the test probiotic group than it did in the control basal diet group (Tufarelli et al., 2017)

    Respiratory effects of low versus high tidal volume with or without positive end-expiratory pressure in anesthetized dogs with healthy lungs

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    OBJECTIVE To evaluate the impact of 2 tidal volumes (TVs) with or without positive end-expiratory pressure (PEEP) on lung mechanics, aeration, and gas exchange in healthy anesthetized dogs. ANIMALS 40 mixed-breed dogs with healthy lungs. PROCEDURES Anesthetized dogs were randomly assigned to 4 groups (n = 10/group) with different ventilatory settings: TVof 8 mL/kg and PEEP of 0 cm H2O (low TVgroup), TVof 8 mL/kg and PEEP of 5 cm H2O (low TVplus PEEP group), TVof 15 mL/kg and PEEP of 0 cm H2O (high TVgroup), or TVof 15 mL/kg and PEEP of 5 cm H2O (high TVplus PEEP group). Expired CO2and respiratory rate were titrated on the basis of a predetermined stepwise protocol. Gas exchange, respiratory mechanics, and pulmonary aeration were evaluated by means of CT 30 minutes after starting mechanical ventilation at the assigned setting. RESULTS Partial pressures of arterial and expired CO2were higher in the low TVand low TVplus PEEP groups than in the high TVand high TVplus PEEP groups. Peak and plateau airway pressures were higher in the PEEP group than in the other groups. Static lung compliance was higher in the high TVplus PEEP group than in the low TVgroup. Relative percentages of atelectatic and poorly aerated lung were lower in the high TVplus PEEP group than in the other groups. Oxygenation was similar among groups. CONCLUSIONS AND CLINICAL RELEVANCE Differences in TVand PEEP application during mechanical ventilation may affect respiratory function in anesthetized dogs with healthy lungs. Ventilation with a TVof 15 mL/kg and PEEP of 5 cm H2O significantly improved lung compliance and reduced the amount of atelectatic and poorly aerated lung

    Engraftment, neuroglial transdifferentiation and behavioral recovery after complete spinal cord transection in rats

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    Background: Proof of the efficacy and safety of a xenogeneic mesenchymal stem cell (MSCs) transplant for spinal cord injury (SCI) may theoretically widen the spectrum of possible grafts for neuroregeneration. Methods: Twenty rats were submitted to complete spinal cord transection. Ovine bone marrow MSCs, retrovirally transfected with red fluorescent protein and not previously induced for neuroglial differentiation, were applied in 10 study rats (MSCG). Fibrin glue was injected in 10 control rats (FGG). All rats were evaluated on a weekly basis and scored using the Basso-Beattie-Bresnahan (BBB) locomotor scale for 10 weeks, when the collected data were statistically analyzed. The spinal cords were then harvested and analyzed with light microscopy, immunohistochemistry, and immunofluorescence. Results: Ovine MSCs culture showed positivity for Nestin. MSCG had a significant and durable recovery of motor functions (P <.001). Red fluorescence was found at the injury sites in MSCG. Positivity for Nestin, tubulin βIII, NG2 glia, neuron-specific enolase, vimentin, and 200 kD neurofilament were also found at the same sites. Conclusions: Xenogeneic ovine bone marrow MSCs proved capable of engrafting into the injured rat spinal cord. Transdifferentiation into a neuroglial phenotype was able to support partial functional recovery

    An underrated complication of the organs' transplantations

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    Tacrolimus is routinely used to prevent rejection after organs' transplantation. Neurotoxicity is underrated side effect, where no typical clinical, radiological, or histopathological patterns have yet been found. The present study is targeted to a review of the literature on tacrolimus-induced neurotoxicity secondary to organs' transplantation, aimed to its prompt diagnosis

    Retrospective study of lameness in beef cattle in northeastern Sardinia, Italy.

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    Lameness is one of the most prevalent diseases affecting dairy and beef cattle, resulting in decreased animal performance, decreased animal welfare, and substantial economic loss. In extensive beef cattle farming, the risk factors for this multifactorial disease are largely unexplored. This study aims to conduct a preliminary epidemiological survey of risk factors in beef cattle in extensive breeding, evaluate the farmer's perception of lameness, and determine the recurrence frequency of the pathologies under investigation in treated animals. The study was conducted in Sardinia, Italy. The population of the study consisted of 14379 cattle from 230 farms. An ad hoc questionnaire was developed to collect all the necessary data. A strong association was found between breed and the occurrence and recurrence of lameness (p < 0.0001). In addition, the Country of origin of both bulls and cows was found to be correlated with the incidence of lameness (p < 0.0001 and 0.0001, respectively). Farmers who indicated on the questionnaire that lameness was not important on their farm had more animals with recurrences (p < 0.0001) than other farmers. The veterinarian's treatment choice differed significantly by farmer concern (p = 0.007) and was associated with less disease recurrence (p < 0.0001), resulting in greater farmer satisfaction (p < 0.007). Pure cow breed, French bull origin, and farmer's age were detected as significant predictors of lameness issues, with pure cow breed and French bull origin having the strongest associations (p = 0.009). Even though the results of this study are preliminary, they indicate that breed selection is crucial in extensive beef farms to reduce lameness prevalence. In addition, it would be reasonable to train breeders to prevent and diagnose lameness early in order to collaborate with veterinarians to prevent recurrence
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