Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial‐like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD‐MSCs) to characterize and differentiate them into endothelial‐like cells. AD‐MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony‐forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM‐2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial‐like cells were characterized by the evaluation of morphological changes and gene expression analysis for endothelial markers (CD31, CD144, CD146). Characterization of AD‐MSCs showed their ability to form clones, to differentiate in vitro and the OCT‐4, SOX‐2, NANOG genes expression. Immunophenotypic characterization showed the CD90 presence and the CD45 absence. The endothelial‐like cells showed morphological changes, the expression of CD31, CD144, CD146 genes and the presence of CD31 membrane receptor. Matrigel assay showed their ability to form network and vessels‐like structures. This study lays the foundations for future evaluation of the potential AD‐MSCs pro‐angiogenic and therapeutic role.     

Foramen magnum decompression with cranioplasty for treatment of caudal occipital malformation syndrome in dogs

Objectives— To describe a cranioplasty procedure used in conjunction with foramen magnum decompression (FMD) for the treatment of canine caudal occipital malformation syndrome (COMS), and to evaluate the clinical outcome. Study Design— Prospective clinical study. Animals— Dogs (n=21) with COMS diagnosed by magnetic resonance imaging (MRI). Methods— After FMD, titanium screws were placed around the perimeter of the foramen magnum defect and a skull plate fashioned from titanium mesh and polymethylmethacrylate was attached to the back of the skull, using the titanium screws as anchor posts. Follow‐up was obtained by direct examination by the authors, telephone interviews with owners and referring veterinarians, and a questionnaire sent to owners of surviving dogs designed to assign objective measures of response to surgical intervention. Surgical success was defined as improvement in ≥1 aspects of clinical dysfunction (e.g. scratching, pain) postoperatively. Owner‐assigned pre‐ and postoperative quality‐of‐life (QOL) scores (1–5) for surviving dogs were compared using a Wilcoxon's signed rank test for paired data (P≤.05). Results— No intraoperative complications occurred and postoperative complications were limited to transient worsening of a pre‐existing head tilt and ataxia in 1 dog, and the need for oral pain medication for 1–4 weeks in 3 dogs. Seventeen dogs (81%) had clinical improvement after surgery. Although clinical signs resolved in 1 dog, it died after being hit by a car, 5 months after surgery. One dog had no change in clinical signs; this patient's clinical signs were not severe and the owners had opted for surgery primarily to prevent progression of disease. Two dogs were euthanatized; 1 because of no improvement of severe clinical signs, and 1 because of sustained recurrence of apparent head and neck pain 7 months later. One dog required a dorsal laminectomy at a second site (C1/C2 vertebral level), 2 months later. Four dogs has transient “flare‐ups” of apparent head/neck pain 2–13 months later; all resolved with transient re‐institution of oral analgesic therapy. Further surgery at the FMD was not needed. Based on the returned questionnaires, there was an overall significant improvement in QOL scores. Conclusions— FMD with cranioplasty was well tolerated, with no intraoperative complications, and minor postoperative complications. Most dogs improved clinically, and none required further surgery at the original FMD site. Clinical Relevance— FMD with cranioplasty may be an effective surgical method of treating dogs with COMS and is well tolerated. Based on prior reports of FMD for this disorder, it appears that cranioplasty may reduce the rate of surgical failure caused by formation of compressive scar tissue at the FMD site in the short term. Results of this preliminary study support further evaluation of the cranioplasty procedure in a larger group of dogs over a more extended follow‐up period.     

Marine and Semi-Synthetic Hydroxysteroids as New Scaffolds for Pregnane X Receptor Modulation

In recent years many sterols with unusual structures and promising biological profiles have been identified from marine sources. Here we report the isolation of a series of 24-alkylated-hydroxysteroids from the soft coral Sinularia kavarattiensis, acting as pregnane X receptor (PXR) modulators. Starting from this scaffold a number of derivatives were prepared and evaluated for their ability to activate the PXR by assessing transactivation and quantifying gene expression. Our study reveals that ergost-5-en-3β-ol (4) induces PXR transactivation in HepG2 cells and stimulates the expression of the PXR target gene CYP3A4. To shed light on the molecular basis of the interaction between these ligands and PXR, we investigated, through docking simulations, the binding mechanism of the most potent compound of the series, 4, to the PXR. Our findings provide useful functional and structural information to guide further investigations and drug design.     

The pharmacokinetic behaviour of marbofloxacin in Eurasian buzzards (Buteo buteo) after intraosseous administration

This study reports on the administration of a single dose of marbofloxacin (2 mg/kg) to five adult Eurasian buzzards (Buteo buteo) by the intraosseous (IO) route, which has been proposed as a rapid and efficient means for the parenteral delivery of antimicrobial drugs. The drug was rapidly absorbed. Peak marbofloxacin concentration (C(max)) in plasma and area under the concentration-time curve (AUC) of 1.92+/-0.78 microg/mL and 8.53+/-2.73 microg h/mL, respectively. The time marbofloxacin remained in the plasma after IO administration was relatively short (elimination half-life, t(1/2beta)=4.91+/-0.65 h; mean residence time (MRT)=5.38+/-0.57 h). Single dose marbofloxacin gave values for C(max)/minimum inhibitory concentration (MIC) of 19.2 and an AUC/MIC value of 85.3h after IO administration. The IO route appears to be practical and effective for the rapid delivery of marbofloxacin to buzzards.