Influence of culture conditions on the secretome of mesenchymal stem cells derived from feline adipose tissue: Proteomics approach.

This study aimed to describe the secretome of mesenchymal stem cells derived from feline adipose tissue (AD-MSCs) and compare the effects of different culture conditions on AD-MSC proteomics using a shotgun approach. Adipose tissue was collected from 5 female cats and prepared to culture. Conditioned media was collected at third passage, in which the cells were cultured under 4 conditions, normoxia with fetal bovine serum (N + FBS), hypoxia with FBS (H + FBS), normoxia without FBS (N - FBS), and hypoxia without FBS (H - FBS). Then, the secretome was concentrated and prepared for proteomic approaches. Secretomes cultured with FBS-free medium had more than twice identified proteins in comparison with the secretomes cultured with FBS. In contrast, hypoxic conditions did not increase protein amount and affected only a small proteome fraction. Relevant proteins were related to the extracellular matrix promoting environmental modulation, influencing cell signaling pathways, and providing a suitable environment for cell proliferation and maintenance. Moreover, other proteins were also related to cell adhesion, migration and morphogenesis. Culture conditions can influence protein abundance in AD-MSC secretome, and can give also more specificity to cell and cell-free treatments for different diseases.

Cross comparison of seminal plasma proteins from cattle and buffalo (Bubalus bubalis).

The objective of this study was to evaluate seminal plasma proteins from cattle and buffalo (Bubalus bubalis), to identify differences between related species. Sixteen buffaloes and 16 cattle between 30 and 60 months of age were used. Semen collection was performed by electroejaculation, followed by macroscopic and microscopic subjective analyses. After analysis, the samples were centrifuged at 800 g for 10 min, and the supernatant (seminal plasma) was recentrifuged at 10,000 g for 30 min at 4°C. The total protein concentration was determined by the Bradford method, and the proteins were digested in solution for mass spectrometry (nLC-MS/MS). Multivariate statistical analysis was used to evaluate the proteomics results by non-hierarchical clustering the considering exponentially modified protein abundance index (emPAI). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used for clustering. Proteomics identified 78 proteins, and multivariate analysis showed 4 that were over-expressed in buffaloes (cystatin C, prosaposin, peptide YY and keratin type II cytoskeletal 5) and 9 in cattle (spermadhesin-1, seminal plasma protein PDC-109, ribonuclease 4, metalloproteinase inhibitor 2, acrosin inhibitor 1, seminal ribonuclease, C-type natriuretic peptide, angiogenin-1 and osteopontin). Among the proteins identified in seminal plasma, the C-type natriuretic peptide and metalloproteinase inhibitors were described for the first time in buffaloes. Some protease inhibitors were found over-expressed in buffaloes, and important proteins in seminal plasma of cattle were not identified or were found at lower expression levels in buffaloes, which can contribute to reproductive performance in this species.

Tratamento medicamentoso de ulceração corneana e abscesso estromal em equinos / Medicament Treatment of Corneal Ulcer and Stromal Abscess in Horses

Background: The equine eyelids and cilia protect the eye and promote normal ocular physiological function. Disorders in these structures may result in corneal alterations such as ulcers and stromal abscess. An abscess is a focal mixture of cellular debris, necrotic tissue, leucocytes and possibly infectious agents that initially create a lesion surrounded by a zone of reactive infl ammation. Ulcers and corneal stromal abscess cause severe ocular pain and loss of vision quality in horses. Medical and surgical treatments for stromal abscess have been developed and have evolved over many years. The aim of this work is to report a case of stromal abscess in a horse, after a corneal ulcer related to eyelid deficiency, focusing on therapeutic methods. Case: A 10-year-old crossbreed mare was referred to the Veterinary Clinics Hospital (HCV) of UFPel with an ophthalmic disorder. The mare had an eyelid lesion 2 years before, and since this event she had had recurrent episodes of ophthalmologic disorders. On attendance at the HCV, the mare had photophobia, blepharospasm, conjunctival hyperemia, neovascularization, myosis, uveitis, intense ocular pain, eyelid deficiency and a corneal ulcer about 2 cm long x 2 cm wide in the right eye. Surgical correction of eyelid and corneal disorder was recommended, but not authorized by the owner. Treatment was based on cleaning with physiological solution, gentamicin eyewash, autologus serum every 4 h, atropine 1% every 12 h topically, flunixin meglumin every 12 h intravenously. After 7 days, the corneal ulcer shrank, although there was no improvement in blepharospasm and ocular pain. A corneal abscess 3 cm x 2 cm was noticed. Gentamicin eyewash and autologus serum were administered topically every 4 h for 10 more days. After 14 days of treatment, there was neither blepharospasm nor corneal ulcer and the abscess was shrinking. Gentamicin eyewash was maintained every 6h for 3 more days. After 65 days there was minimal scar tissue in the eye. Discussion: The mare had an ocular traumatic injury 2 years ago and since this event she had presented uveitis and corneal ulcer recurrently, due to loss of eyelid and cilia functional integrity. Equine cornea is avascularized and vascularization of the affected lesion is necessary for resolution and abscess healing. Medical treatment of superfi cial stromal abscess is more economical than surgery and can result in good visual outcomes with mild scarring. Medical treatment consists of a combination of antimicrobial, mydriatic/cycloplegic and anti-inflammatory medications. Flunixin meglumin seems to be the most effective intravenous anti-inflammatory for controlling uveitis in horses. The anti-inflammatory dose may be reduced when there is a decrease in the anterior segment signs of infl ammation and ocular pain. Besides systemic colateral effects, anti-inflammatory therapy also reduces corneal vascularization, and therefore should be used carefully. Antibiotic topical therapy is highly recommended, due to ocular microflora and microorganism encapsulation, and it was used in the present report until there was full epithelial healing. A mydriatic/cycloplegic (atropine 1%) drug was administered aiming to dilate the pupil in order to prevent adherences and to reduce ocular discomfort. Antiproteases agents (autologus serum) are recommended when there is a corneal epithelial loss of integrity. This agent reduces collagen degradation and helps corneal healing. Stromal abscess in horses is usually associated with previous corneal ulceration, and in the present report it also involved eyelid functional deficiency. Medical treatment was effective, ensuring quality of future vision for the animal.