A crescente poluição ambiental tem mobilizado atitudes no sentido de promover mudanças nos hábitos individuais e coletivos, que refletirão na construção de um novo cenário, pautado no desenvolvimento sustentável. Alternativas para preservação do meio ambiente podem minimizar problemas de saúde enfrentados atualmente e melhorar a qualidade de vida das futuras gerações. Entre os resíduos domésticos e de estabelecimentos alimentícios, os óleos de frituras destacam-se pelos danos causados quando lançados diretamente nas tubulações, poluindo os rios e dificultando o tratamento nas estações de tratamento de água. Neste trabalho, desenvolvido como projeto de extensão, propõe-se transformar o óleo usado em sabão (processo conhecido como saponificação) em barra ou líquido, através de procedimentos práticos caseiros e de baixo custo, como uma alternativa de preservação ambiental. Os sabões produzidos tiveram suas propriedades monitoras, como a capacidade de espumar, o odor e o potencial hidrogeniônico (pH). Os procedimentos eram abordados através da realização de palestras e oficinas, bem como pela divulgação dos procedimentos através de folders e cartilhas. A análise dos questionários aplicados durante as atividades desenvolvidas entre agosto de 2007 e julho de 2008 permitiu concluir que 82% dos participantes das oficinas lançavam os óleos usados diretamente nas tubulações e que 69% expressaram comprometimento de que continuariam reciclando a partir da participação nas oficinas. Estima-se que um público maior foi atingido devido aos procedimentos terem sido divulgados na TV local.
The aims of this study were to verify the effects of Epidermal Growth Factor (EGF) on the morphology, primordial follicle activation, growth and proliferation of granulosa cells of ovine follicles cultured in situ, as well as the effect of a PI3K inhibitor on the follicular activation. Ten ovine ovaries were divided into fragments, being one fixed for histological analysis (fresh control). The remaining fragments were cultured for 7 days in control medium (α-MEM + ) alone or supplemented with EGF (1, 10, 50, 100 or 200 ng/mL). Follicles were classified as normal or atretic, as primordial or growing, and the oocyte and follicle diameters were measured. PCNA immunohistochemistry was performed in the fresh control and in treatment that showed the best results for follicular activation. Pharmacologic inhibition of PI3K activity was performed through pretreatment in media added with 50 µM LY294002 for 1 h. The percentage of normal follicles decreased (P < 0.05) after 7 days of culture in all treatments compared to the fresh control. A significant reduction in the percentage of primordial follicles and an increase (P < 0.05) in the growing ones were observed in all treatments compared to fresh control. Furthermore, both the control medium and 1 ng/mL EGF promoted an increase (P < 0.05) in follicular activation compared to other EGF treatments. The PCNA-positive cells in the EGF treatment were higher (P < 0.05) than in fresh control and α-MEM + . Pretreatment of ovarian tissue with PI3K inhibitor significantly inhibited (P < 0.05) α-MEM + -stimulated primordial follicle activation, but had no effect on EGF-stimulated activation (P > 0.05). In conclusion, PI3K pathway mediates the in vitro spontaneous activation of sheep primordial follicles. Moreover, EGF may act indirectly on follicular activation by promoting granulosa cell proliferation at 1 ng/mL, and EGF inhibited follicle activation in concentrations similar or higher than 10 ng/mL.
The aim of this study was to verify the beneficial effects of different concentrations of epidermal growth factor (EGF) on survival and antrum formation of isolated ovine preantral follicles cultured in vitro. Ovine ovaries (n = 50) were collected from a local slaughterhouse and secondary follicles (150–200 μm in diameter), without antral cavities, were mechanically isolated by microdissection using 26-gauge needles. After selection, the follicles were individually cultured in 100-μL droplets of culture medium at 39°C and 5% CO2 in air for 18 days. The basic control medium consisted of α-minimal essential medium (α-MEM) supplemented with BSA; insulin, transferring and selenium; glutamine; hypoxanthine; and ascorbic acid and then referred to as α-MEM+. For the experimental conditions, follicles were cultured in α-MEM+ alone (control) or in different concentrations of EGF (1, 10, or 50 ng mL–1). Every other day, 60 μL of the culture media was replaced with fresh media. The morphological aspects of all ovine follicles were assessed every 6 days using a precalibrated ocular micrometer in a stereomicroscope at 100× magnification. Only those follicles showing an intact basement membrane, with bright and homogeneous granulosa cells and an absence of morphological signs of degeneration, were classified as morphologically normal follicles. The rupture of the basement membrane was also observed and characterised as follicle extrusion. In addition, antral cavity formation was defined as the emergence of a visible translucent cavity within the granulosa cell layers. Data from morphologically normal follicles, extruded follicles, and antrum formation rate during in vitro culture were expressed as percentages and compared by the chi-squared test, and differences were considered significant when P < 0.05. The results showed that the percentage of morphologically normal follicles decreased significantly throughout the culture periods in all the treatments, except in the 50 ng mL–1 EGF group, which maintained the percentage of normal follicles from Day 0 to 6. Considering the same culture period, 50 ng mL–1 EGF treatment significantly increased the percentage of morphologically normal follicles at Day 18 compared with the control group. Moreover, the addition of EGF to the culture medium, at 50 ng mL–1, significantly reduced the precocious extrusion of oocytes and increased the percentage of antrum formation compared with the control and 1 ng mL–1 EGF after 18 days of culture. Notably, there were no significant differences between 10 ng mL–1 EGF, control medium, and 1 ng mL–1 EGF treatments. In conclusion, this study demonstrated that the addition of EGF to the in vitro culture medium, at 50 ng mL–1, increased the proportion of morphologically normal follicles and antrum formation rate of isolated ovine preantral follicles. This work was supported by FACEPE (Process APQ-0705–5.05/10). L. P. Santos is a recipient of a grant from FACEPE (Brazil).
It was possible to develop a stable experimental model of saccular aneurysms in pig carotid artery by use of the internal jugular vein.
Article historyThis study evaluated the effect of caprine ovarian tissue transportation conditions (medium supplementation and transportation duration) on the morphology, DNA fragmentation and development of cultured and non-cultured preantral follicles. After the fragmentation of ovaries, one fragment was fixed (fresh control) while the remaining slices were placed individually in two different conservation media (Minimal Essential Medium -MEM without supplementation or supplemented MEM, i.e. MEM + ) and stored at 35ºC for 6 or 12 h without (non-cultured) or with a subsequent 5-day in vitro culture in supplemented α-MEM. After transportation, followed or not by in vitro culture, the fragments were processed for histological and Terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) examination. For the preserved and non-cultured fragments, the percentages of normal follicles after the storage of ovarian tissue in MEM + for 6 h and the DNA fragmentation rates after preservation in MEM for 6 h and MEM + for 6 or 12 h were maintained similar to the fresh control. However, all cultured treatments reduced the proportion of normal follicles and increased the percentage of TUNEL-positive cells as compared to the fresh control and non-cultured treatments. On the contrary, all culture conditions (except after preservation in MEM for 6 h) promoted an increase in primordial follicle activation. In conclusion, the use of an enriched medium (MEM + ) during ovary transportation is preferable to maintain satisfactory rates of normal follicles after the preservation of caprine ovarian tissue at 35ºC for up to 6 h, without affecting the ability of the primordial follicle to grow in vitro.
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