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Immunophenotypic characterisation and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture
2016
Mazurkevych, Anatoliy | Malyuk, Mykola | Bezdieniezhnykh, Natalia | Starodub, Lyubov | Kharkevych, Yuriy | Brusko, Evgen | Gryzińska, Magdalena | Jakubczak, Andrzej
Introduction: The objective of the study was immunophenotypic and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture.Material and Methods: The mesenchymal stem cells were obtained from equine bone marrow of three horses and from foal umbilical cords of six foals. The cells were cultured in CO₂ incubators by standard procedure. Quantitative abnormalities of chromosomes, i.e. aneuploidy and polyploidy, and structural aberrations, i.e. breaks in chromosomes and chromatid, were taken into account during the study.Results: The results of cytogenetic analysis of equine bone marrow mesenchymal stem cells at the third and fourth passages indicated that the level of karyotype variability of these cells corresponded to the spontaneous level of karyotype variability typical of the peripheral blood lymphocytes of this species. Equine bone marrow contained several clones of stem cells that differed in the expression of specific nuclear markers characteristic of proliferating cells.Conclusion: Mesenchymal stem cells from foal umbilical cords during in vitro cultivation are characterised by quantitative abnormalities of the chromosomal apparatus.
اظهر المزيد [+] اقل [-]Evaluation of the role of the cyclooxygenase signaling pathway during inflammation in skin and muscle tissues of ball pythons (Python regius)
2016
Sadler, Ryan A. | Schumacher, Juergen P. | Rathore, Kusum | Newkirk, Kim M. | Cole, Grayson | Seibert, Rachel | Cekanova, Maria
OBJECTIVE To determine degrees of production of cyclooxygenase (COX)-1 and -2 and other mediators of inflammation in noninflamed and inflamed skin and muscle tissues in ball pythons (Python regius). ANIMALS 6 healthy adult male ball pythons. PROCEDURES Biopsy specimens of noninflamed skin and muscle tissue were collected from anesthetized snakes on day 0. A 2-cm skin and muscle incision was then made 5 cm distal to the biopsy sites with a CO2 laser to induce inflammation. On day 7, biopsy specimens of skin and muscle tissues were collected from the incision sites. Inflamed and noninflamed tissue specimens were evaluated for production of COX-1, COX-2, phosphorylated protein kinase B (AKT), total AKT, nuclear factor κ-light-chain-enhancer of activated B cells, phosphorylated extracellular receptor kinases (ERKs) 1 and 2, and total ERK proteins by western blot analysis. Histologic evaluation was performed on H&E-stained tissue sections. RESULTS All biopsy specimens of inflamed skin and muscle tissues had higher histologic inflammation scores than did specimens of noninflamed tissue. Inflamed skin specimens had significantly greater production of COX-1 and phosphorylated ERK than did noninflamed skin specimens. Inflamed muscle specimens had significantly greater production of phosphorylated ERK and phosphorylated AKT, significantly lower production of COX-1, and no difference in production of COX-2, compared with production in noninflamed muscle specimens. CONCLUSIONS AND CLINICAL RELEVANCE Production of COX-1, but not COX-2, was significantly greater in inflamed versus noninflamed skin specimens from ball pythons. Additional research into the reptilian COX signaling pathway is warranted.
اظهر المزيد [+] اقل [-]Evaluation of epiduroscopy for detection of vertebral canal and spinal cord lesions in dogs
2016
Garcia-Pereira, Fernando L. | Prange, Timo | Seller, Aaron | Obert, Vicotria
OBJECTIVE To evaluate the potential usefulness of epiduroscopy for clinical diagnosis and treatment of vertebral canal and spinal cord lesions in dogs. SAMPLE Cadavers of 6 mixed-breed dogs. PROCEDURES Dogs were positioned in sternal recumbency, and an endoscope was introduced into the lumbosacral epidural space. A fiberscope (diameter, 0.9 mm; length, 30 cm) was used for 3 dogs, and a videoscope (diameter, 2.8 mm; length, 70 cm) was used for the other 3 dogs. Visibility and identities of anatomic structures were recorded, and maneuverability of the endoscopes was assessed. Extent of macroscopic tissue damage was evaluated by manual dissection of the vertebral canal at the end of the procedure. RESULTS Intermittent saline (0.9% NaCl) solution infusion, CO2 insufflation, and endoscope navigation improved visualization by separating the epidural fat from the anatomic structures of interest. Images obtained with the fiberscope were small and of poor quality, making identification of specific structures difficult. Maneuverability of the fiberscope was difficult, and target structures could not be reliably reached or identified. Maneuverability and image quality of the videoscope were superior, and spinal nerve roots, spinal dura mater, epidural fat, and blood vessels could be identified. Subsequent manual dissection of the vertebral canal revealed no gross damage in the spinal cord, nerve roots, or blood vessels. CONCLUSIONS AND CLINICAL RELEVANCE A 2.8-mm videoscope was successfully used to perform epiduroscopy through the lumbosacral space in canine cadavers. Additional refinement and evaluation of the technique in live dogs is necessary before its use can be recommended for clinical situations.
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