Not Always Toxoplasmosis: Active Retinochoroiditis Presumably Secondary to Herpes in Eyes with Hyperpigmented Retinochoroidal Previous Scars.

PURPOSE: To demonstrate that the presence of active retinochoroiditis in eyes with previous hyperpigmented old retinochoroidal lesions is not exclusive of ocular toxoplasmosis. METHODS: A case series was constructed by reviewing medical records. RESULTS: Four Brazilian patients presenting active posterior uveitis in eyes with previous hyperpigmented old retinochoroidal lesions were identified. Ocular toxoplasmosis was ruled out in all cases. One case had viral etiology confirmed through vitreous PCR (HSV-2 was positive), and the other 3 cases had a presumed diagnosis of herpetic posterior uveitis. CONCLUSION: Focus active retinitis adjacent to an old cicatricial lesion should not be considered pathognomonic of toxoplasmosis and viral etiology must be considered.

Arthroscopic Disc Suturing and Repositioning Technique for Treating Anteriorly Displaced Disc in Temporomandibular Joint Disorders: A Posterior Anchorage Technique.

Several treatment modalities have been reported for patients with arthrogenous temporomandibular disorders. The most common temporomandibular disorder is internal derangement, usually accompanied by displacement of the articular disc. Most often anteriorly and medially displaced, patients may develop clicking, impaired jaw function, and pain. Articular disc repositioning has been described as an effective method to eliminate interference during mandibular translation, improving mandibular range of motion, and eliminating pain. In this article, we present a new, simple, and reproducible technique without requiring specific instruments for repositioning and suturing the articular disc with posterior anchorage fixed to the tragus cartilage. It has the advantages of a better anteroposterior vector of traction force, without adverse effects as lateralization of the articular disc and skin depression, commonly presented in other techniques.

Recurrence of Ocular Toxoplasmosis after Vitrectomy: Case Report and Review.

PURPOSE: The purpose of this study is to report one case of ocular toxoplasmosis (OT) recurrence after vitrectomy and review the scientific basis about it. CASE REPORT: A 58-year-old male patient with previous OT, properly treated, underwent vitrectomy due to macular hole. During follow-up, patient evolved with recurrence of the OT. After 1 year, patient presents visual acuity of 20/200 and extensive macular scar. CONCLUSION: There is no consensus on using perioperative antiparasitic therapy aiming recurrence prophylaxis. Studies with better statistical design are necessary to evaluate the recurrence risk after ocular surgeries and the possible recommendation of prophylaxis, especially in countries where the strains are more virulent and the recurrence more common.

Avaliação do cuidado hospitalar pelo risco de óbito / Evaluation of hospital care by risk of death

A taxa de mortalidade hospitalar é tradicionalmente usada para medir a qualidade do cuidado nas unidades hospitalares, no entanto a mesma não consegue discriminar o risco de óbito proveniente da assistência oferecida e da carga de comorbidades que o paciente tem no momento da internação. A Razão de Mortalidade Hospitalar Padronizada (RMHP) é um índice que possibilita avaliar a qualidade do cuidado de acordo com o perfil dos pacientes atendidos pelo hospital. Resultados da RMHP < 1 indica uma qualidade melhor que a esperada e resultados > 1 apontam qualidade pior que a esperada. Assim este estudo propôs avaliar a qualidade da assistência hospitalar prestada no âmbito da Secretaria de Estado da Saúde de Goiás, considerando seu perfil assistencial e os tipos de pacientes atendidos, no período de 2014 a 2019. O cálculo da RMHP foi baseado na metodologia de Jarman et al., no Índice de Comorbidades de Charlson (ICC) E Índice de Elixhauser para avaliar o risco de óbito, e posterior aplicação da curva ROC (para encontrar o modelo de ajuste de risco) e da regressão linear logística multivariada. A qualidade da assistência prestada no Estado de Goiás, considerando o perfil dos pacientes atendidos, foi pior que o esperado nos anos de 2014 a 2016, dentro do esperado no ano 2017 e melhor que o esperado nos anos de 2018 e 2019. Observou-se uma melhora gradativa na qualidade do cuidado nos últimos três anos

The hospital mortality rate is traditionally used to measure the quality of care in hospital units, however it cannot discriminate the risk of death resulting from the assistance provided and the burden of comorbidities that the patient has at the time of hospitalization. The Standardized Hospital Mortality Ratio (RMHP) is an index that makes it possible to assess the quality of care according to the profile of patients treated by the hospital. RMHP results < 1 indicate better quality than expected and results > 1 indicate worse quality than expected. Thus, this study proposed to assess the quality of hospital care provided by the State Health Department of Goiás, considering its care profile and the types of patients cared for, in the period from 2014 to 2019. The RMHP calculation was based on Jarman's methodology et al., in the Charlson Comorbidity Index (ICC) and Elixhauser Index to assess the risk of death, and subsequent application of the ROC curve (to find the risk adjustment model) and multivariate logistic linear regression. The quality of care provided in the State of Goiás, considering the profile of the patients cared for, was worse than expected in the years 2014 to 2016, within expectations in 2017 and better than expected in the years 2018 and 2019. A gradual improvement in the quality of care in the last three years

Questões étnico-raciais, educação física escolar e educação para o lazer / The financing of leisure in Brazil by the different federated entities

Este artigo tem como objetivo analisar o tema questões étnico-raciais e Educação Física escolar e apresentar apontamentos sobre a educação para o lazer. Desde a promulgação da lei 10.639/2003, com o reconhecimento da necessidade da presença da história e cultura afro-brasileira nos currículos escolares, tem sido um importante passo para o reforço do trabalho pedagógico com as questões étnico-raciais na Educação Física. Como procedimentos metodológicos foi realizada pesquisa bibliográfica. Foram selecionados textos com data de publicação de 2016 a 2020. A análise é de natureza qualitativa. As questões étnico-raciais trabalhadas na Educação Física escolar com foco na educação para o lazer são uma oportunidade para a discussão/vivência sobre práticas de diversas culturas e etnias e sua valorização. Conclui-se que este pode ser um caminho efetivo para minimizar problemas sociais como o preconceito.

This paper aims at analyzing ethnic-racial issues and school Physical Education as well as presenting notes on leisure for education. Since the enactment of Law 10.639 / 2003, with the recognition of the need for the presence of Afro-Brazilian history and culture in school curricula, it has been an important step towards strengthening the pedagogical work with ethnic-racial issues in Physical Education. Methodological procedures were performed by bibliographical research. Texts with publication data from 2016 to 2020 were selected. This is a qualitative analysis. Ethnicracial issues addressed in school Physical Education with a focus on education for leisure are an opportunity for discussion/experience on practices of different cultures and ethnicities and their appreciation. We concluded that this can be an effective way to minimize social problems such as prejudice.

Antineoplastic activity of a novel ruthenium complex against human hepatocellular carcinoma (HepG2) and human cervical adenocarcinoma (HeLa) cells.

Novel metal complexes have received much attention recently because of their potential anticancer activity. Notably, ruthenium-based complexes have emerged as good alternatives to the currently used platinum-based drugs for cancer therapy, with less toxicity and fewer side effects. The beneficial properties of Ru, which make it a highly promising therapeutic agent, include its variable oxidative states, low toxicity, and high selectivity for cancer cells. The present study evaluated the cytotoxic effects of a ruthenium complex, namely cis-[Ru(1,10-phenanthroline)2(imidazole)2]2+ (RuC), on human hepatocellular carcinoma (HepG2) and human cervical adenocarcinoma (HeLa) cells and analyzed metabolic parameters. RuC reduced HepG2 and HeLa cell viability at all tested concentrations (10, 50, and 100 nmol/L) at 48 h of incubation, based on the MTT, Crystal violet, and neutral red assays. The proliferation capacity of HepG2 cells did not recover, whereas HeLa cell proliferation partially recovered after RuC treatment. RuC also inhibited all states of cell respiration and increased the levels of the metabolites pyruvate and lactate in both cell lines. The cytotoxicity of RuC was higher than cisplatin (positive control) in both lineages. These results indicate that RuC affects metabolic functions that are related to the energy provision and viability of HepG2 and HeLa cells and is a promising candidate for further investigations that utilize models of human cervical adenocarcinoma and mainly hepatocellular carcinoma.

Characterization and Application of Yarrowia lipolytica Lipase Obtained by Solid-State Fermentation in the Synthesis of Different Esters Used in the Food Industry.

Yarrowia lipolytica lipase obtained by solid-state fermentation was characterized and applied in the synthesis of esters with commercial value in the food industry. The effect of different conditions on the hydrolysis activity of this biocatalyst was evaluated in the presence of metal ions, solvents, detergents, several pH and temperature parameters, and different substrates. Storage stability was also studied. The solid biocatalyst produced in soybean meal was used in synthesis reactions aiming to produce short-, medium-, and long-chain esters. Results showed that the best fermentation condition to produce the biocatalyst was using soybean oil (3% w/w), moisture content (55% w/v), and inoculum of 2.1 mgdry biomass/gsoybean meal at 28 °C for 14 h. High substrate conversion for ethyl octanoate, cetyl stearate, and stearyl palmitate synthesis was achieved in the presence of non-polar solvents in less than 6 h using a substrate molar ratio of 1:1 at 38 °C with 10-15% (w/v) of biocatalyst. This work showed the high potential of Y. lipolytica lipase to be used in the synthesis of different esters. Also, that it can be considered an attractive and economical process alternative to obtain high-added value products.

Biocatalytic esterification of fatty acids using a low-cost fermented solid from solid-state fermentation with Yarrowia lipolytica.

This study aimed to evaluate the use of a lyophilized fermented solid (named solid enzymatic preparation, SEP), with lipase activity, as a low-cost biocatalyst for esterification reactions of fatty acids present in acid raw materials for biodiesel synthesis. The SEP was obtained by solid-state fermentation (SSF) of soybean bran using the strain of Yarrowia lipolytica IMUFRJ 50682 and contains the lipases secreted by this yeast. The esterification reaction of ethanol and the predominant fatty acids present in different acid oil sources for biodiesel production (oleic, linoleic, stearic and palmitic acids) was investigated. Oleic acid conversion of above 85% was obtained after 24 h, using 30 wt% of SEP and ethanol/oleic acid molar ratio of 1, at 30 °C, in a reaction medium with and without solvent (n-hexane). Similar results were achieved with stearic (79%), palmitic (82%) and linoleic (90%) acids. The reusability of SEP was investigated over ten successive batches by washing it with different solvents (ethanol, water or n-hexane) between the cycles of ethyl oleate synthesis. Washing with water allowed the SEP to be reused for six cycles maintaining over 80% of the conversion reached in the first cycle. These results show the potential of this biocatalyst to reduce the content of free fatty acids in acid oils for biodiesel synthesis with a potential to be applied in a broad plethora of raw materials.

Effects of silymarin on angiogenesis and oxidative stress in streptozotocin-induced diabetes in mice.

The present study evaluated the effects of acute treatment with silymarin, an extract that is obtained from Silybum marianum, on angiogenesis, oxidative stress, and inflammation in normoglycemic and diabetic mice. Diabetes was induced by streptozotocin (80 mg/kg, intraperitoneal) in male Swiss mice, 6 weeks of age. A polyether-polyurethane sponge was surgically implanted in the back of the mice as a model of healing in both diabetic and normoglycemic animals that were treated with oral silymarin or water for 10 days. The pancreas, liver, kidneys, blood, and sponges were collected and analyzed. Diabetes led to impairments of antioxidant defenses, reflected by a reduction of pancreatic superoxide dismutase and hepatic and renal catalase and an increase in pancreatic lipoperoxidation. An inflammatory process was observed in diabetic mice, reflected by an increase in pancreatic tumor necrosis factor α (TNF-α) and the infiltration of inflammatory cells in islets. The number of vessels was lower in the implanted sponges in diabetic mice. Silymarin treatment attenuated this damage, restoring antioxidant enzymes and reducing pancreatic TNF-α and inflammatory infiltration. However, silymarin treatment did not restore angiogenesis or glycemia. In conclusion, treatment with silymarin red uced oxidative stress and inflammation that were induced in the model of streptozotocin-induced diabetes in several organs, without apparent toxicity. Silymarin may be a promising drug for controlling diabetic complications.

Associação da escoliose toracolombar e peso da mochila em estudantes do ensino médio / Association between thoracolumbar scoliosis and backpack weight in high school students

Introdução: O aparecimento das alterações posturais em es-colares pode estar relacionada à prática de hábitos posturais inadequados, como o transporte de peso excessivo nas mochilas escolares. Objetivo:Verificar se existe associação entre a utilização de mochilas pelos estudantes do ensino médio com escoliose. Casuística e Métodos:Trata-se de uma pesquisa descritiva trans-versal com abordagem quantitativa, com 152 estudantes do en-sino médio de duas escolas, que responderam a um formulário de coleta de dados e foram submetidos à avaliação postural por inspeção estática e fotogrametria. Resultados: A escoliose toracolombar estrutural foi a alteração postural mais frequente identificada após realização do teste de Adams(68%). Hou-ve associação entre a presença de escoliose e gênero feminino (p=0,02), peso da mochila maior que 10% do peso do estu-dante (p= 0,01) e tipo de mochila unilateral (p=0,003). Conclusão: Houve associação estatisticamente significante entre a escoliose toracolombar e o peso mochila maior que 10% da massa corpórea na amostra analisada, além do predomínio de mochila unilateral e gênero feminino.

Introduction:The presence of postural changes in schoolchildren may be related to inappropriate posture habits, such as excessive weight transport in school backpacks.Objective: To verify if there is an association between the uses of backpacks by high school students and scoliosis. Patients and Methods:This is a cross-sectional descriptive research with a quantitative approach, with 152 high school students from two schools, who responded to a data collection form and were submitted to postural evaluation by static inspection and photogrammetry. Results:Structural thoracolumbar scoliosis was the most frequent postural change identified after the Adams test (68%). Results also shown that there was an association between the presence of scoliosis and female students (p=0.02). We found the backpack weight greater than 10% of student weight (p=0.01), and a backpack positioned unilaterally on the back (p=0.003). Conclusion: There was a statistically significant association between thoracolumbar scoliosis and backpack weight greater than 10% of the body mass in the analyzed sample, besides an association between backpack positioned unilaterally on the back and female students.

Ruthenium complex exerts antineoplastic effects that are mediated by oxidative stress without inducing toxicity in Walker-256 tumor-bearing rats.

The present study evaluated the in vivo antitumor effects and toxicity of a new Ru(II) compound, cis-(Ru[phen]2[ImH]2)2+ (also called RuphenImH [RuC]), against Walker-256 carcinosarcoma in rats. After subcutaneous inoculation of Walker-256 cells in the right pelvic limb, male Wistar rats received 5 or 10mgkg-1 RuC orally or intraperitoneally (i.p.) every 3 days for 13 days. A positive control group (2mgkg-1 cisplatin) and negative control group (vehicle) were also used. Tumor progression was checked daily. After treatment, tumor weight, plasma biochemistry, hematology, oxidative stress, histology, and tumor cell respiration were evaluated. RuC was effective against tumors when administered i.p. but not orally. The highest i.p. dose of RuC (10mgkg-1) significantly reduced tumor volume and weight, induced oxidative stress in tumor tissue, reduced the respiration of tumor cells, and induced necrosis but did not induce apoptosis in the tumor. No clinical signs of toxicity or death were observed in tumor-bearing or healthy rats that were treated with RuC. These results suggest that RuC has antitumor activity through the modulation of oxidative stress and impairment of oxidative phosphorylation, thus promoting Walker-256 cell death without causing systemic toxicity. These effects make RuC a promising anticancer drug for clinical evaluation.

Aspiration and cytological evaluation of idiopathic bone cavities of the jaw.

The idiopathic bone cavity (IBC) is an intraosseous pseudocyst devoid of epithelial lining. Clinically, IBCs of the jaw are asymptomatic and normally found in routine radiographic exams. Although the literature regarding the content of IBCs is controversial, the final diagnosis is usually aided by the discovery of an empty cavity upon surgical exploration. The aim of this study was to perform cytological and histological analysis of IBC contents. Cytological analysis of nine cases of IBC was performed after puncture and processed by the cell block technique. Histological analysis was performed in six cases in which it was possible to collect enough material by curettage of bone walls. Remarkably, cell block analysis revealed the presence of fibrin, often arranged as a net; erythrocytes; and inflammatory cells, with a predominance of lymphocytes as well as some macrophages and neutrophils. Histological analysis showed the presence of scant connective tissue, bone trabeculae, hemorrhagic foci, and hemosiderin. Only two cases presented scattered multinucleated giant cells. Cytological evaluation of IBC content by the cell block technique might represent a useful diagnostic tool, especially in cases in which there is no available material for curettage in the cavity.

Could BDNF be involved in compensatory mechanisms to maintain cognitive performance despite acute sleep deprivation? An exploratory study.

BACKGROUND: Neuroimaging studies suggest that acute sleep deprivation can lead to adaptations, such as compensatory recruitment of cerebral structures, to maintain cognitive performance despite sleep loss. However, the understanding of the neurochemical alterations related to these adaptations remains incomplete. OBJECTIVE: Investigate BDNF levels, cognitive performance and their relations in healthy subjects after acute sleep deprivation. METHODS: Nineteen sleep deprived (22.11±3.21years) and twenty control (25.10±4.42years) subjects completed depression, anxiety and sleep quality questionnaires. Sleep deprived group spent a full night awake performing different playful activities to keep themselves from sleeping. Attention, response inhibition capacity and working memory (prefrontal cortex-dependent) were assessed with Stroop and Digit Span tests. Declarative memory (hippocampus-dependent) was assessed with Logical Memory test. Serum BDNF was measured by sandwich ELISA. Data were analyzed with independent samples T-test, ANOVA, ANCOVA and curve estimation regressions. p<0.05 was deemed statistically significant. RESULTS: The sleep deprived group showed higher BDNF levels and normal performance on attention, response inhibition capacity and working memory. However, declarative memory was impaired. A sigmoidal relation between BDNF and Stroop Test scores was found. CONCLUSIONS: Increased BDNF could be related, at least in part, to the maintenance of normal prefrontal cognitive functions after sleep deprivation. This potential relation should be further investigated.

Evaluation of two formulations containing mineral trioxide aggregate on delayed tooth replantation: relevance of RANKL/RANK/OPG system.

This study aimed to evaluate the effects of White MTA (WMTA) and MTA Fillapex(®) on root resorption, when used for root canal filling, in a rat model of delayed tooth replantation, with special focus on the RANKL/RANK/OPG system. Maxillary right central incisors of male rats were extracted (total N = 48), and exposed to dry environment for 30 min. The animals were allocated into four groups: (1) WMTA; (2) MTA Fillapex; (3) Calcium hydroxide; (4) Negative control. After periodontal ligament removal, root canals were filled with the corresponding material and replanted. After 10 and 60 days, qualitative and semi-quantitative histological and immunohistochemical analyses were carried out. Analysis of variance (ANOVA) with Tukey's post hoc adjustment was used, at 10 and 60 days, to compare the experimental groups in terms of the inflammatory scores and in terms of the changes in OPG, RANK and RANKL. Both WMTA and MTA Fillapex groups displayed inflammatory and replacement resorption, with the presence of dento-alveolar ankylosis, similarly to that observed for calcium hydroxide, in either 10 or 60 days. Notably, a slight increase of the inflammatory process was observed in both MTA groups. Quantitatively, inflammation score analysis showed a significant difference between the calcium hydroxide and the control group at 10 days. On 60 days, dento-alveolar ankylosis was found significantly increased in the MTA Fillapex, in comparison to the control group (p < 0.05). For immunohistochemical analysis, the expression of both RANK and RANKL was reduced in calcium hydroxide and WMTA groups, from 10 to 60 days of evaluation, an effect that was accompanied by increased OPG immunolabelling. Otherwise, the MTA Fillapex group presented a general increase of RANKL immunopositivity, similarly to that observed in the negative control group. Our data showed that none of tested materials was able to fully prevent the root resorption, although the white MTA cement presented an outcome comparable to that seen for calcium hydroxide. MTA cements might present some advantages when considering no need of frequent changes, although the effects of MTA cements in dental avulsion still require further investigation.

Proteome of Soybean Seed Exudates Contains Plant Defense-Related Proteins Active against the Root-Knot Nematode Meloidogyne incognita.

Several studies have described the effects of seed exudates against microorganisms, but only few of them have investigated the proteins that have defensive activity particularly against nematode parasites. This study focused on the proteins released in the exudates of soybean seeds and evaluated their nematicidal properties against Meloidogyne incognita. A proteomic approach indicated the existence of 63 exuded proteins, including ß-1,3-glucanase, chitinase, lectin, trypsin inhibitor, and lipoxygenase, all of which are related to plant defense. The presence of some of these proteins was confirmed by their in vitro activity. The soybean exudates were able to reduce the hatching of nematode eggs and to cause 100% mortality of second-stage juveniles (J2). The pretreatment of J2 with these exudates resulted in a 90% reduction of the gall number in tobacco plants. These findings suggest that the exuded proteins are directly involved in plant defense against soil pathogens, including nematodes, during seed germination.

Selective Cytotoxicity of 1,3,4-Thiadiazolium Mesoionic Derivatives on Hepatocarcinoma Cells (HepG2).

In this work, we evaluated the cytotoxicity of mesoionic 4-phenyl-5-(2-Y, 4-X or 4-X-cinnamoyl)-1,3,4-thiadiazolium-2-phenylamine chloride derivatives (MI-J: X=OH, Y=H; MI-D: X=NO2, Y=H; MI-4F: X=F, Y=H; MI-2,4diF: X=Y=F) on human hepatocellular carcinoma (HepG2), and non-tumor cells (rat hepatocytes) for comparison. MI-J, M-4F and MI-2,4diF reduced HepG2 viability by ~ 50% at 25 µM after 24-h treatment, whereas MI-D required a 50 µM concentration, as shown by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The cytotoxicity was confirmed with lactate dehydrogenase assay, of which activity was increased by 55, 24 and 16% for MI-J, MI-4F and MI-2,4diF respectively (at 25 µM after 24 h). To identify the death pathway related to cytotoxicity, the HepG2 cells treated by mesoionic compounds were labeled with both annexin V and PI, and analyzed by flow cytometry. All compounds increased the number of doubly-stained cells at 25 µM after 24 h: by 76% for MI-J, 25% for MI-4F and MI-2,4diF, and 11% for MI-D. It was also verified that increased DNA fragmentation occurred upon MI-J, MI-4F and MI-2,4diF treatments (by 12%, 9% and 8%, respectively, at 25 µM after 24 h). These compounds were only weakly, or not at all, transported by the main multidrug transporters, P-glycoprotein, ABCG2 and MRP1, and were able to slightly inhibit their drug-transport activity. It may be concluded that 1,3,4-thiadiazolium compounds, especially the hydroxy derivative MI-J, constitute promising candidates for future investigations on in-vivo treatment of hepatocellular carcinoma.

Characterization of an alcoholic hepatic steatosis model induced by ethanol and high-fat diet in rats

Alcoholic liver disease is characterized by a wide spectrum of liver damage, which increases when ethanol is associated with high-fat diets (HFD). This work aimed to establish a model of alcoholic hepatic steatosis (AHS) by using a combination of 10% ethanol and sunflower seeds as the source of HFD. Male rats received water or 10% ethanol and regular chow diet and/or HFD, which consisted of sunflower seeds. The food consumption, liquid intake and body weight of the rats were monitored for 30 days. After this period, blood was collected for biochemical evaluation, and liver samples were collected for histological, mitochondrial enzyme activity and oxidative stress analyses. Our results indicated that the combination of 10% ethanol and HFD induced micro- and macrosteatosis and hepatocyte tumefaction, decreased the levels of reduced glutathione and glutathione S-transferase activity and increased the level of lipoperoxidation and superoxide dismutase activity. The mitochondrial oxidation of NADH and succinate were partially inhibited. Complexes I and II were the main inhibition sites. Hepatic steatosis was successfully induced after 4 weeks of the diet, and the liver function was modified. The combination of 10% ethanol and sunflower seeds as an HFD produced an inexpensive model to study AHS in rats.

Vitrified sheep isolated secondary follicles are able to grow and form antrum after a short period of in vitro culture.

The risk of reintroducing malignant cells after ovarian graft into patients following post-cancer treatment is an obstacle for clinical applications (autotransplantation). In this context, in vitro follicle culture would be an alternative to transplantation in order to minimize such risks. Therefore, the aim of this study was to compare the development of secondary follicles after vitrification in isolated form (without stroma) with vitrification in in situ form (within fragments of ovarian tissue). Follicles were first isolated from ovarian fragments from mixed-breed ewes and then vitrified; these comprised the Follicle-Vitrification group (Follicle-Vit), or fragments of ovarian tissue were first vitrified, followed by isolation of the follicles, resulting in the Tissue-Vitrification group (Tissue-Vit). Control and vitrified groups were submitted to in vitro culture (6 days) and follicular morphology, viability, antrum formation, follicle and oocyte diameter, growth rate, ultrastructural characteristics and cell proliferation were evaluated. The percentages of morphologically normal follicles and antrum formation were similar among groups. Follicular viability and oocyte diameter were similar between Follicle-Vit and Tissue-Vit. The follicular diameter and growth rate of Follicle-Vit were similar to the Control, while those of Tissue-Vit were significantly lower compared to the Control. Both vitrified groups had an augmented rate of granulosa cellular proliferation compared to Control. Secondary follicles can be successfully vitrified before or after isolation from the ovarian tissue without impairing their ability to survive and grow during in vitro culture.

Activin-A promotes the development of goat isolated secondary follicles in vitro.

The role of activin-A in follicular development and on the mRNA expression levels of different genes in goat secondary follicles was evaluated. Goat secondary follicles (≥ 150 µm) were cultured for 18 days under control conditions or with the addition of either 50 or 100 ng/ml activin-A (Experiment 1). The mRNA levels for the genes that code for activin-A, ActR-IA, ActR-IB, ActR-IIA, ActR-IIB, follicle stimulating hormone receptor (FSH-R) and P450 aromatase were measured in each condition (Experiment 2). We observed that after 6 days of culture, the antrum formation rate was higher in cultures with added activin-A than in the cultured control (P < 0.05). The addition of 50 ng/ml activin-A increased the follicular growth rate in the final third of the culture (days 12-18), resulting in a higher percentage of meiosis resumption (P < 0.05). On day 6, the addition of activin-A (50 ng/ml) increased the levels of ActR-IA mRNA compared with the cultured control (P < 0.05). After 18 days, the addition of 50 ng/ml activin-A significantly increased the levels of its own mRNA compared with the non-cultured control. Moreover, this treatment reduced the mRNA levels of P450 aromatase in comparison with the cultured control (P < 0.05). Higher levels of P450 aromatase mRNA were found for both activin-A treatments compared with the non-cultured control (P < 0.05). No difference in estradiol levels was detected among any of the tested treatments. In conclusion, the addition of activin-A to culture medium stimulated early antrum formation as well as an increase in the daily follicular growth rate and the percentage of meiosis resumption.