Effects of orexin B on swine granulosa and endothelial cells

In addition to the well-known central modulatory role of orexins, we recently demonstrated a peripheral involvement in swine granulosa cells for orexin A and in adipose tissue for orexin B (OXB). The aim of present research was to verify immunolocalization of OXB and its potential role in modulating the main features of swine granulosa cells. In particular, we explored the effects on granulosa cell proliferation (through the incorporation of bromodeoxyuridine), cell metabolic activity (as indirect evaluation by the assessment of ATP), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test and the non-enzymatic reducing power by FRAP test). Our data point out that OXB does not modify granulosa cell growth, steroidogenesis and superoxide anion generation. On the contrary, the peptide stimulates (p < 0.05) nitric oxide output and non-enzymatic reducing power. Since new vessel growth is crucial for ovarian follicle development, a further aim of this study was to explore the expression of prepro-orexin and the effects of OXB on swine aortic endothelial cells. We found that the peptide is ineffective in modulating cell growth, while it inhibits redox status parameters. In addition, we demonstrated a stimulatory effect on angiogenesis evaluated in fibrin gel angiogenesis assay. Taken together, OXB appears to be potentially involved in the modulation of redox status in granulosa and endothelial cells and we could argue an involvement of the peptide in the follicular angiogenic event

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products

Multifocal cutaneous non-epitheliotropic B-cell lymphoma in a cat

Case summary Skin tumours are the second-most common form of feline cancer after haematopoietic neoplasms and are often malignant. Cutaneous lymphoma is uncommon in cats and can be classified as epitheliotropic (typically of T-cell origin) or non-epitheliotropic (either of T-cell or B-cell origin). The present study describes a case of multifocal cutaneous non-epitheliotropic B-cell lymphoma. The skin nodules were multiple and variable in size; showed rapid progression; were alopecic and erythematous in appearance and pruritic and ulcerated; and were mostly located on the trunk. Nodule biopsies revealed the presence of uniform medium-to-large round neoplastic cells that infiltrated the dermis and subcutis. The neoplasias were consistent with a round cell cutaneous tumour and did not show evidence of epitheliotropism. Furthermore, immunohistochemical assessments indicated an immunophenotype characterised by round cells with a strong membrane and cytoplasmic positivity for the CD20 antigen, consistent with a lymphocyte of B-cell origin. Relevance and novel information Cutaneous non-epitheliotropic B-cell lymphoma in cats is rare and was previously reported to appear as single dermal and subcutaneous masses that are variable in size and generally develop in the tarsal region. To our knowledge, this is the first report to describe multifocal cutaneous non-epitheliotropic B-cell lymphoma in a cat

Are virgin olive oils obtained below 27 C better than those produced at higher temperatures?

a b s t r a c t Within the European Union, indications of 'first cold pressing' and 'cold extraction' can only be used for virgin olive oil (VOO) obtained below 27 C from mechanical processing. Three different malaxing temperatures (25, 35 and 45 C) are here evaluated for the quality of the VOO obtained in a continuous industrial plant. The oils were stored at room temperature in the dark for 12 months. Initially, oil obtained from a blend of Frantoio/Leccino cultivars (F/L) had higher acidity and peroxide levels and lower phenolic content than a Coratina cultivar (Cor). The oxidative stability of the oils positively correlated with malaxation temperature (F/L, R 2 ¼ 0.818; Cor, R 2 ¼ 0.987) as the phenolic content was directly proportional to the temperature (F/L, R 2 ¼ 0.887; Cor, R 2 ¼ 0.992). Only oils obtained at 45 C were rejected because of 'heated or burnt' off-flavour. Decarboxymethylation of secoiridoids and further hydrolysis of phenolic esters occurred during storage. The oxidation products of derivatives of tyrosol and hydroxytyrosol were detected after nine months in both the F/L and Cor samples. Thus, VOO obtained at a processing temperature lower than 27 C does not show higher chemical and sensory qualities than VOO obtained at 35 C

PRELIMINARY EVALUATION OF IMMUNOHISTOCHEMICAL EXPRESSION OF INHIBIN-α AND GATA-4 IN NORMAL AND NEOPLASTIC CANINE GONADS

Introduction: Inhibin-α is a protein produced by gonadal stromal cells responsible for the physiological regulation of the productive axis in both genders, and is connected to mechanisms of cell differentiation and tumor suppression. GATA-4 is a transcription factor which is differentially expressed depending on gonadal developmental stages and pathological conditions. Materials and Methods: Immunohistochemistry for Inhibin-α and GATA-4 was performed on normal gonads of both genders at different developmental stages and also on neoplasms. Immunostaining was evaluated with a semiquantitative score. Results: Inhibin-α was expressed in Sertoli cells of normal testis of prepubertal dogs. Meanwhile in mature dogs, Leydig cells were also positive. In both sex-cord stromal tumors (Leydig and Sertoli cell tumors) neoplastic cells were positive. In normal ovary, follicular epithelial cells of primordial follicles and granulosa cells of remnant follicle stages were positive. Granulosa cell tumors expressed Inhibin-α in neoplastic cells, meanwhile papillary cystadenomas were negative. GATA-4 was expressed in normal testis of prepubertal and pubertal dogs in Sertoli and Leydig cells. In both sex-cord stromal tumors, neoplastic cells were positive. In normal ovary, prepubertal dogs had positive stromal and epithelial cells. In pubertal and mature females, granulosa cells were positive with viable expression in stromal cells. Granulosa cell tumors expressed GATA-4 in neoplastic cells and papillary cystadenoma was negative. Conclusions: Inhibin-α can be used as marker of normal and neoplastic sex-cord stromal cells in testis and ovarian granulosa cells. GATA-4 has a similar expression in testis and a constant expression in normal and neoplastic ovarian granulosa cells

Nematodi bronco-polmonari del gatto: rilievi anatomoistopatologici in tre gatti del nord Italia

SUMMARY Aelurostrongylus abstrusus is regarded as the major lungworm infecting Felis catus, although other poorly studied, nematodes have been described from the respiratory system of domestic cats. The present paper describes the postmortem macroscopic lesions and the histopathological observations of the infection by metastrongyloid nematodes in three cats in Northern ltal

Melatonin modulates swine luteal and adipose stromal cell functions

Based on our previous study in follicle, the first aim of the work was to evaluate the effect of melatonin in the swine corpus luteum. Luteal cells were exposed to 10 and 20 pg mL-1 melatonin. We evaluated the effect on proliferation (Bromo-deoxy-Uridine uptake), steroidogenesis (progesterone) and redox status by means of Griess test (nitric oxide production), WST-1 test (superoxide anion generation) and FRAP test (non-enzymatic antioxidant power). The results show a significant increase of the antioxidant power as well as a reduction of the other parameters analyzed. These data and the expression of MT2 observed in luteal cells, allow us to hypothesize a physiological role of melatonin in the regulation of corpus luteum functionality. The reproductive function is dependent on energy reserves stored in adipose tissue. Therefore, we sought to verify effect of melatonin on adipose stromal cells (ASCs). MT2 receptor expression was detected in ASCs and the presence of gene markers (PPARγ and leptin) before and after adipogenic differentiation was verified. The differentiation was significantly inhibited by melatonin, as well as cell viability. In conclusion, present results suggest that melatonin exerts a potential inhibitory action on luteal function and adipogenesis, possibly mediated by MT2