RESUMO
Osteoarthritis (OA) is one of the main locomotor disorders in horses. Although nonsteroidal anti-inflammatory drugs are the first-line treatment for OA, opioids could also be used. In previous studies, opioids showed promising anti-inflammatory and analgesic effects. In this study, we aimed to investigate the effects of two opioids (morphine and methadone) against inflammation in lipopolysaccharide (LPS)-stimulated synoviocytes by analyzing microsomal prostaglandin E synthase-1 (mPGES-1) and prostaglandin-endoperoxide synthase 2 (PTGS2) expression. Synoviocytes were obtained from the joints at the distal limbs of dead animals. The cytotoxic effects of LPS, morphine, and methadone were investigated by using a cell viability assay with crystal violet dye. Synoviocytes were treated with LPS, LPS plus morphine, or LPS plus methadone for 3, 6, and 12â¯h, and mPGES-1 and PTGS2 expression was measured using real-time polymerase chain reaction. LPS, and morphine did not affect the viability of synoviocytes, even at high concentrations. LPS treatment increased mPGES-1 and PTGS2 expression, whereas morphine inhibited the increase in mPGES-1 and PTGS2 expression in LPS-stimulated synoviocytes. Methadone did not inhibit mPGES-1 or PTGS2 expression. These results suggest that morphine may exhibit anti-inflammatory effect; therefore, it might be beneficial for the treatment of OA.
Assuntos
Ciclo-Oxigenase 2/química , Inflamação/tratamento farmacológico , Lipopolissacarídeos/toxicidade , Metadona/farmacologia , Microssomos/enzimologia , Morfina/farmacologia , Prostaglandina-E Sintases/antagonistas & inibidores , Sinoviócitos/efeitos dos fármacos , Analgésicos Opioides/farmacologia , Animais , Células Cultivadas , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Cavalos , Inflamação/induzido quimicamente , Inflamação/enzimologia , Masculino , Sinoviócitos/imunologia , Sinoviócitos/metabolismo , Sinoviócitos/patologiaRESUMO
The sprawl of the urbanization and road network process without building ecological corridors contributes to the high mortality rates and a threat to the population decline of wild species such as the crab-eating fox. A strategy for the ex situ conservation is the study of the reproductive biology of the species and cryopreservation of their genetic heritage through the formation of an animal germplasm bank. This research is in accordance with the principles adopted by Brazilian College of Animal Experimentation. Reproductive systems of Cerdocyon thous females (n = 7) were examined macroscopically and microscopically by histological techniques and scanning electron microscopy. Gross features showed the shape of the ovaries was similar to a bean, and the elongated oviducts lengths were between 5 and 8 cm, with body of the uterus (3 cm) with long and narrow uterine horns (9-11 cm). The cervix was as a single annular conformation carrying out communication between the uterus and the vagina. The vagina has lengthened and circular muscle and the vulva with dense anatomical conformation with a quite pronounced clitoris. In addition, with regard to the establishment of a cell line (fibroblasts) for the gene bank enrichment, cells showed a low clonogenic capacity, especially when compared to domestic dogs, which can be explained by "in vitro" environment, age and diet of the animal. However, it was possible to create a bank of limited cell number. This study had morphological and preservationist character and aimed to help at long term in the conservation of wild animal's genetic resources.
Assuntos
Biodiversidade , Bancos de Espécimes Biológicos , Canidae/anatomia & histologia , Criopreservação/veterinária , Genitália Feminina/anatomia & histologia , Animais , Brasil , Canidae/genética , Meios de Cultura , Feminino , Fibroblastos/citologia , Fibroblastos/ultraestrutura , Genitália Feminina/ultraestrutura , Masculino , Microscopia Eletrônica de Varredura/veterináriaRESUMO
Takahashi and Yamanaka established the first technique in which transcription factors related to pluripotency are incorporated into the genome of somatic cells to enable reprogramming of these cells. The expression of these transcription factors enables a differentiated somatic cell to reverse its phenotype to an embryonic state, generating induced pluripotent stem cells (iPSCs). iPSCs from canine fetal fibroblasts were produced through lentiviral polycistronic human and mouse vectors (hOSKM/mOSKM), aiming to obtain pluripotent stem cells with similar features to embryonic stem cells (ESC) in this animal model. The cell lines obtained in this study were independent of LIF or any other supplemental inhibitors, resistant to enzymatic procedure (TrypLE Express Enzyme), and dependent on bFGF. Clonal lines were obtained from slightly different protocols with maximum reprogramming efficiency of 0.001%. All colonies were positive for alkaline phosphatase, embryoid body formation, and spontaneous differentiation and expressed high levels of endogenous OCT4 and SOX2. Canine iPSCs developed tumors at 120 days post-injection in vivo. Preliminary chromosomal evaluations were performed by FISH hybridization, revealing no chromosomal abnormality. To the best of our knowledge, this report is the first to describe the ability to reprogram canine somatic cells via lentiviral vectors without supplementation and with resistance to enzymatic action, thereby demonstrating the pluripotency of these cell lines.
Assuntos
Feto/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/fisiologia , Fator Inibidor de Leucemia/farmacologia , Células-Tronco Pluripotentes/fisiologia , Animais , Cães , Fibroblastos/citologia , Regulação da Expressão Gênica/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Reação em Cadeia da Polimerase/veterináriaRESUMO
Very few carnivore's embryology is reported mainly restricted to old literature without new technique analyses. Also, their development focuses on pharyngeal arches and stem cell sources and the high capacity for differentiation from those cells to generate embryonic tissue. We aimed to use immunohistochemistry to prove the potentiality of these stem cell niches. The results were to highlight the timetable for the development of dogs and cats, the proper formation of pharyngeal arches and the description of these cells on first and second arches since 17-25 days of pregnancy. After that, the differentiation process is reduced.