Bilateral malignant seminoma with metastases in the mule: a report of two cases.

We report two cases of bilateral malignant seminoma in the bursula testium of two mules, which had been slaughtered normally. Histologically, the testicular tumours consisted predominantly of large round to polyhedral cells with frequent mitotic figures. Metastases were found in the sublumbar lymph nodes. A final histopathological diagnosis of malignant seminoma with metastasis was made. To our knowledge, this paper appears to be the first report of two cases of bilateral malignant seminoma with metastases, in the mule.     

Duplex real-time polymerase chain reaction for simultaneous detection and quantification of Anaplasma marginale and Anaplasma centrale

Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 10(1) and 10(2) DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.     

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.     

In vitro fermentation of feed ingredients by fresh or frozen pig fecal inocula

In vitro techniques can offer a rapid, repeatable and cost‐effective alternative to in vivo experiments. We investigated the gas production (GP) of nine different feeds commonly used in Italian and Maltese pig farms in order to assess the use of the Ankom system in this species and to compare the in vitro microbial activity fresh or frozen inocula derived from piglet feces. Proximate and polyphenolic analyses were determined for all substrates. Fermentation profiles were measured according to the cumulative GP technique. Analysis of GP values revealed significant differences in the fermentation patterns mainly due to substrate and inoculum effects. Fermentation parameters varied significantly according to the substrate ingredients. The frozen inoculum generally led to more total GP than for fresh, with the exception of hard wheat bran pellets (HWBP), pelleted pulp and mature carob. Conversely the fresh fecal inoculum had an earlier maximum time to reach maximum GP than the frozen, showing a higher maximum rate of GP in all substrates excepting for HWBP. A significant difference in frozen and fresh inocula was found for the complete formulation. Further studies are required to confirm the variation between fresh and frozen fecal inocula to a wider range of feed, including those that could have negative biological effects.     

Anaerobic Digestion of Olive Oil Mill Effluents: Evaluation of Wastewater Organic Load and Phytotoxicity Reduction

Olive oil mill effluents (OME) are of great concern worldwide dueto their role as pollutants. This work studies, on a laboratoryscale, the possibility of reducing the polluting load of olivewastewaters (organic, acid and polyphenolic) through chemical-physical processes (sedimentation-filtration) followedby a biological treatment (anaerobic digestion). This is in orderto produce biogas and obtain an effluent suitable for applicationto the soil. The anaerobic process was followed by determiningchemical-physical parameters (pH, COD, N-NH₃, polyphenols).The extent of anaerobic digestion was evaluated throughmeasurements of biogas and volatile fatty acids and the microbialmetabolism was examined through dehydrogenase activity. Finally,tests of plant germination and growth were carried out usingdigested waters to determine if they could be used in agriculture.The study demonstrated that anaerobic digestion of OME firstreduced the organic load by 78-89% and the content of polyphenolsby 33-43%, and secondly produced biogas (mean value of methane83-85%). Phytotoxicity tests carried out on Lepidiumsativum seeds showed that the anaerobic treatment considerablyreduced the phyto-toxic character of OME.     

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.     

Effects of Genotype and Environment on Phenolic Acids Content and Total Antioxidant Capacity in Durum Wheat

In cereals, phenolic acid (PA) content and total antioxidant capacity (TAC) may have a wide range of variability, probably because of several factors influencing the occurrence of grain antioxidants, which include genotype, environment, and their possible interactions. However, only a few studies have investigated the influence of these factors on durum wheat. In the present study, we investigated the impact of the genetic and environmental factors on the profile and content of PAs occurring as soluble free, soluble conjugated, and insoluble bound compounds, as well as on the TAC level, in three genotypes of durum wheat grown in three different Italian agroclimatic areas during two crop years. The results show that genotype, environment, and crop year have highly significant effects on TAC levels and on PA contents. In particular, TAC and free PAs are most influenced by year, whereas conjugated and bound PAs are most influenced by environment × year and genotype, respectively. Therefore, it is evidenced that genetic and environmental factors affect the antioxidant activity and the content of the three forms of PAs in durum wheat to different extents.