Aloe vera increases mRNA expression of antioxidant enzymes in cryopreserved bovine ovarian tissue and promotes follicular growth and survival after in vitro culture.

The aims of the present study were to evaluate the effects of Aloe vera extract on expression of mRNA for antioxidant enzymes in bovine ovarian tissue after vitrification, as well as on follicular morphology, viability, activation and extracellular matrix in cultured ovarian tissues that had been previously vitrified. Fragments from bovine ovarian cortical tissue were cryopreserved in a vitrification solution alone or supplemented with two concentrations of Aloe vera (10 or 50%). After thawing, the cryopreserved tissues were analyzed by histological techniques, as well as the levels of mRNA for SOD, CAT, PRDX6 and GPX1 were investigated. Furthermore, cryopreserved fragments were then culture in vitro in α-MEM for 6 days. Histological evaluation of cultured tissues was performed to determine the percentages of normal and developing follicles. The results showed that, after vitrification, the presence of Aloe vera in both concentrations was able to maintain percentages of collagen fibers similar to fresh tissues (P < 0.05). Aloe vera in both concentrations significantly increased mRNA levels for PRDX6 and GPX1 in cryopreserved tissues, while 10% Aloe vera increased mRNA levels for SOD (P < 0.05). In parallel, after in vitro culture, fragments vitrified in the presence of 10% Aloe vera had significantly higher levels of morphologically healthy follicles when compared to tissue that were vitrified without Aloe vera. In fragments vitrified with Aloe vera, the rate of developing follicles was significantly higher than in tissues vitrified without Aloe vera. Tissues vitrified with 10% Aloe vera and cultured in vitro maintained percentages of collagen fibers similar to fresh tissues. In conclusion, 10% Aloe vera increases the expression of mRNA for PRDX6, GPX1 and SOD in vitrified ovarian tissues, maintains follicular survival and promotes activation and development of follicles after in vitro culture of vitrified bovine ovarian tissue.

Effects of vulvar width and antral follicle count on oocyte quality, in vitro embryo production and pregnancy rate in Bos taurus taurus and Bos taurus indicus cows.

It was hypothesized that Brahman cows have greater in vitro embryo production than Simmental cows, and that regardless of breed, antral follicle count (AFC) and vulvar width (VW) are positively associated with oocyte quality, embryo number and pregnancy rate. Brahman (Bos taurus indicus, n = 184) and Simmental (Bos taurus taurus, n = 99) cows were classified as having a large AFC (Brahman: ≥50, Simmental: ≥25); intermediate AFC (Brahman: >30 and ≤49; Simmental: >16 and <24) and small AFC (Brahman: <30, Simmental: ≤15). The VW was considered to be large (Brahman: >123 mm, Simmental: >80 mm); intermediate (Brahman: >102 mm and ≤123 mm; Simmental: >65 mm and ≤80 mm) and small (Brahman: ≤102 mm and Simmental: ≤65 mm). For each group of cows, ovum pick up, oocyte in vitro maturation, fertilization and embryo culture were performed. Embryo transfers were subsequently performed and pregnancy rate was evaluated. Brahman cows had a larger number of viable oocytes, greater efficiency of embryo production and greater pregnancy rate (P < 0.05) than Simmental cows. Pregnancy percentages in Simmental cows with a small AFC were greater than those cows with an intermediate and large AFC. With the Brahman cows, there was no difference in pregnancy rate among the cows with different AFC classifications. In contrast to Simmental cows, Brahman cows with an intermediate VW had a greater pregnancy percentage than those with a small and large AFC. In conclusion, Brahman cows have a larger AFC, number of viable oocytes, and pregnancy rate than Simmental cows.

RT-qPCR study of COX-1 and -2 genes in oral surgical model comparing single-dose preemptive ibuprofen and etoricoxib: A randomized clinical trialy.

BACKGROUND: This study aimed to evaluate the gene expression of cyclooxygenases (COXs) in an oral model of preemptive analgesia. MATERIAL AND METHODS: Gingival tissue was collected during extraction of lower third molars from a randomized, triple-blind, split-mouth and placebo-controlled study. The eligible patients were randomly sorted to receive a single dose either of ibuprofen 400mg, or etoricoxib 120 mg or a placebo, one hour prior to surgery. The temporal course of RNAm was evaluated for COX-1 and -2 by means of a quantitative polymerase chain reaction in real time (RT-qPCR) at time zero and 30 minutes after the surgical procedure began, and it was correlated with clinical parameters (pain and maximum mouth opening). RESULTS: There was a significant increase in COX-1 expression between T0 and T30 in ibuprofen (p=0.004) and etoricoxib (p=0.010) groups. As regards COX-2, there were increases from T0 to T30 in all groups (placebo, p=0.012; ibuprofen, p<0.001; etoricoxib, p<0.001). All groups showed a significant decrease in COX-2:COX-1 ratio from T0 to T30 (placebo, p=0.013; ibuprofen, p<0.001; etoricoxib, p=0.047). Experimental groups showed a significant correlation between COX-1 and COX-2 levels and clinical pain parameters. CONCLUSIONS: The present preemptive analgesia study concludes that COX-2 RNAm induction was directly linked to third molar-related tissue inflammation and that the relation between COX-1 and COX-2 levels were inversely proportional to the preemptively administered nonsteroidal anti-inflammatory drugs COX-2 selectivity. Key words:Preemptive analgesia, dental extraction, cyclooxygenases, real-time polymerase chain reaction.

Ultrastructure of Sheep Primordial Follicles Cultured in the Presence of Indol Acetic Acid, EGF, and FSH.

The aim of this study was to investigate the ultrastructural characteristics of primordial follicles after culturing of sheep ovarian cortical slices in the presence of indol acetic acid (IAA), Epidermal Growth Factor (EGF), and FSH. To evaluate ultrastructure of primordial follicles cultured in MEM (control) or in MEM containing IAA, EGF, and FSH, fragments of cultured tissue were processes for transmission electron microscopy. Except in the control, primordial follicles cultured in supplemented media for 6 d were ultrastructurally normal. They had oocyte with intact nucleus and the cytoplasm contained heterogeneous-sized lipid droplets and numerous round or elongated mitochondria with intact parallel cristae were observed. Rough endoplasmic reticulum (RER) was rarely found. The granulosa cells cytoplasm contained a great number of mitochondria and abundant RER. In conclusion, the presence of IAA, EGF, and FSH helped to maintain ultrastructural integrity of sheep primordial follicles cultured in vitro.