A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the "Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro" / Um resumo das espécies de répteis e anfíbios anuros das planícies costeiras arenosas brasileiras: 31 anos de esforços de amostragem do "Laboratório de Ecologia de Vertebrados da Universidade do Estado do Rio de Janeiro"

Abstract Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called "restinga" and "campo nativo" habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.

Resumo Embora atualmente exista um conjunto de estudos sobre aspectos ecológicos de algumas espécies de répteis e de anfíbios que ocorrem nas planícies costeiras arenosas brasileiras (incluindo os chamados habitats de "restinga" e de "campo nativo"), há relativamente poucas informações sobre a composição de espécies geralmente associada a esses ambientes. Durante 31 anos (1988-2019) de estudos herpetológicos realizados em restingas por nossa equipe de pesquisa do Laboratório de Ecologia de Vertebrados (Departamento de Ecologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil) nós estudamos comunidades de répteis e de anfíbios e realizamos diferentes estudos com métodos semelhantes em 70 localidades de dez diferentes Estados ao longo da costa brasileira. Nossas pesquisas resultaram em registros de 87 espécies de répteis (cinco tartarugas, dois crocodilianos, seis anfisbênios, 36 lagartos e 39 serpentes) de 24 famílias, e 77 espécies de anfíbios anuros de nove famílias. Estudamos vários tópicos de história natural sobre anuros e répteis, que resultaram na publicação de alguns estudos ecológicos específicos, especialmente em relação a algumas espécies, abrangendo ecologia populacional e de comunidades, forrageamento e dieta, horário de atividade de espécies, termorregulação, reprodução, uso do microhabitat e parasitismo por ecto e endoparasitas. Nossos resultados ao longo dessas três décadas também contribuíram para a descrição de quatro novas espécies de lagartos (Ameivula nativo, Glaucomastix littoralis, G. abaetensis e G. itabaianensis). Nossos estudos constituem uma importante contribuição para o conhecimento da ecologia de répteis e de anfíbios anuros nesses ecossistemas, bem como para a conservação dos ecossistemas de restinga. A lista de espécies apresentada neste estudo, com base em nossos registros de herpetofauna das planícies costeiras arenosas, fornece para muitas localidades ao longo da costa brasileira o conhecimento necessário sobre a ocorrência de espécies, incluindo a presença de espécies endêmicas e/ ou ameaçadas de extinção, que podem ser úteis para muitas ações de conservação.

Do restinga cnemidophorine lizards run on empty along the Brazilian coast? / Os lagartos cnemidophorine de restinga correm no vazio ao longo da costa brasileira?

Abstract The number of individuals with empty stomachs in a population (i.e. the proportion of individuals "running on empty") can be used as a simple index of instantaneous energy balance of some organisms such as lizards and fishes. In this study, we aimed to analyze the proportion of empty stomachs in 16 populations of five cnemidophorine species (Glaucomastix abaetensis, Ameivula ocellifera, Glaucomastix littoralis, Contomastix lacertoides and Ameivula nativo) along approximately 5,000 km the Brazilian coast. A total of 550 individuals had their stomach contents removed and identified in the laboratory. Our results showed that the proportion of individuals "running on empty" varied from 0 to 11.1% among the different populations and species. These proportions are suggestive that the five studied species would be in an overall positive energy balance at the time of the study.

Resumo O número de indivíduos com estômagos vazios em uma população (i.e. a proporção de indivíduos "correndo no vazio") pode ser usado como um índice simples do balanço energético instantâneo de organismos como lagartos e peixes. Este estudo teve como objetivo analisar a proporção de estômagos vazios em 16 populações pertencentes a cinco espécies de cnemidophorines (Glaucomastix abaetensis, Ameivula ocellifera, Glaucomastix littoralis, Contomastix lacertoides e Ameivula nativo) ao longo de cerca de 5,000 km da costa leste do Brasil. Um total de 550 indivíduos tiveram seus conteúdos estomacais removidos e analisados em laboratório. Nossos resultados mostraram que a proporção de indivíduos "correndo no vazio" variou entre 0 a 11.1% entre as diferentes populações e espécies. Essas proporções sugerem que as cinco espécies estudadas estariam em geral em um balanço energético positivo, quando da realização deste estudo.

Leptin decreases apoptosis and promotes the activation of primordial follicles through the phosphatidylinositol-3-kinase/protein kinase B pathway in cultured ovine ovarian tissue.

This study evaluated the effects of leptin on primordial follicle survival and activation after in vitro culture of ovine ovarian tissue and if leptin acts through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway. Ovarian fragments were fixed for histology (fresh control) or cultured for 7 days in control medium (α-MEM+) alone or supplemented with leptin (1, 5, 10, 25 or 50 ng/ml). Follicle morphology, activation and apoptosis were analyzed. Next, the fragments were cultured in the medium that showed the best results in the absence or the presence of the PI3K inhibitor (LY294002), and immunohistostaining of p-Akt protein was assessed. After culture, the percentage of normal follicles decreased (P < 0.05) in all treatments compared with the fresh control. Moreover, control medium and 1 ng/ml leptin had similar (P > 0.05) percentages of normal follicles, which were significantly higher than those in other treatments. However, culture with 1 ng/ml leptin maintained apoptosis similarly (P > 0.05) to that of the fresh control and lower (P < 0.05) than that in α-MEM+. Leptin did not influence follicle activation (P > 0.05) compared with the control medium (α-MEM+). Culture in 1 ng/ml leptin with LY294002 decreased the normal follicles and increased apoptosis, inhibited follicle activation (P < 0.05), and reduced p-Akt immunostaining, compared with the medium containing 1 ng/ml leptin without PI3K inhibitor. In conclusion, leptin at 1 ng/ml reduces apoptosis and promotes the activation of primordial follicles compared with the fresh control after in vitro culture of ovine ovarian tissue possibly through the PI3K/Akt pathway.

Involvement of Phosphorylated Akt and FOXO3a in the Effects of Growth and Differentiation Factor-9 (GDF-9) on Inhibition of Follicular Apoptosis and Induction of Granulosa Cell Proliferation After In Vitro Culture of Sheep Ovarian Tissue.

This study aimed to evaluate the effects of growth and differentiation factor-9 (GDF-9) on the morphology, activation, apoptosis, and granulosa cell proliferation of ovine preantral follicles cultured within ovarian tissue slices and to verify whether GDF-9 could influence follicular activation through the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O3a (PI3K/Akt/FOXO3a) pathway. Ovine ovarian fragments were cultured in α-MEM+ or α-MEM+ with GDF-9 (1, 50, 100, 200, or 400 ng/ml) for 7 days. Apoptosis and cell proliferation were analyzed. Next, the activation of the PI3K was inhibited with LY294002, and immunostaining for p-Akt and p-FOXO3a proteins was assessed. The concentration of 50 ng/ml GDF-9 had (P < 0.05) more morphologically normal follicles compared to all treatments, except 1 ng/ml GDF-9. Moreover, 50 ng/ml GDF-9 increased primordial follicle activation compared to all treatments, except α-MEM+ and 1 ng/ml GDF-9. However, the concentration of 50 ng/ml GDF-9 showed higher cell proliferation and lower apoptosis than α-MEM+ and 1 ng/ml GDF-9 treatments. Culture of the ovarian tissue with LY294002 inhibited the activation of primordial follicles and reduced p-Akt immunostaining in both α-MEM+ and 50 ng/ml GDF-9 treatments. In addition, after culture with LY294002, the percentage of oocytes with nuclear p-FOXO3 was higher in 50 ng/ml GDF-9 than in the control medium (α-MEM+). In conclusion, after culture of ovine ovarian cortical slices, the addition of 50 ng/ml GDF-9 reduces follicular apoptosis and promotes granulosa cell proliferation likely through the involvement of phosphorylated Akt and FOXO3a.

Antimicrobial activity and toxicity of glass ionomer cement containing an essential oil.

The aim of this study was to evaluate the antimicrobial activity and toxicity of glass ionomer cement (GIC) modified with 5-methyl-2-(1-methylethyl)phenol (thymol) against Streptococcus mutans in silico and in vitro. The antimicrobial activity of thymol on GIC modified with concentrations of 2% (GIC-2) and 4% (GIC-4) was evaluated in a model of planktonic cell biofilm using agar diffusion test, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), dynamic biofilm (continuous flow cell parallel), and bacterial kinetics. Conventional GIC (GIC-0) was used as a control. Thymol toxicity was evaluated in Artemia salina and in silico using Osiris® software. Differences between groups were estimated by analysis of variance, followed by Tukey post hoc test, with a 5% significance level. The results of the agar diffusion test between groups were not significantly different (P≥0.05). Thymol had potential bacteriostatic and bactericidal activity against Streptococcus mutans with respect to planktonic growth, with MIC of 100 µg/mL and MBC of 400 µg/mL. The groups GIC-0, GIC-2, and GIC-4 reduced the biofilm by approximately 10, 85, and 95%, respectively. Bacterial kinetics showed efficiency of the modified GICs for up to 96 h. GIC with thymol was effective against S. mutans, with significant inhibition of the biofilms. Analyses in silico and using Artemia salina resulted in no relevant toxicity, suggesting potential for use in humans. GIC-2 was effective against S. mutans biofilm, with decreased cell viability.

Epigallocatechin-3-gallate (EGCG) reduces apoptosis of preantral follicles through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) signaling pathway after in vitro culture of sheep ovarian tissue.

The aims of this study were to analyze the effects of different concentrations of epigallocatechin-3-gallate (EGCG) on the primordial follicle survival and development after in vitro culture of ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) pathway in the EGCG actions in the sheep ovary. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or with different concentrations of EGCG (0.01; 0.1; 1; 10 or 100 µg/mL) for 7 days. Inhibition of PI3K activity was performed in fragments cultured with 1 µg/mL EGCG plus LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 and AKT phosphorylation (p-AKT). The results showed that 1 µg/mL EGCG maintained the follicular survival similar (P > 0.05) to that of the fresh control and higher (P < 0.05) than that of the α-MEM+ and other EGCG treatments. No difference (P > 0.05) in the follicular activation was observed. However, both follicle and oocyte diameters increased after in vitro culture with 1 µg/mL EGCG compared to other treatments (P < 0.05), except for 10 µg/mL EGCG (P > 0.05). After PI3K inhibition, there was an increase (P < 0.05) of the follicular apoptosis and a reduction of p-AKT immunolocalization. In conclusion, EGCG at 1 µg/mL reduces apoptosis of preantral follicles through the PI3K/AKT pathway after in vitro culture of sheep ovarian tissue.

Antimicrobial activity and toxicity of glass ionomer cement containing an essential oil

The aim of this study was to evaluate the antimicrobial activity and toxicity of glass ionomer cement (GIC) modified with 5-methyl-2-(1-methylethyl)phenol (thymol) against Streptococcus mutans in silico and in vitro. The antimicrobial activity of thymol on GIC modified with concentrations of 2% (GIC-2) and 4% (GIC-4) was evaluated in a model of planktonic cell biofilm using agar diffusion test, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), dynamic biofilm (continuous flow cell parallel), and bacterial kinetics. Conventional GIC (GIC-0) was used as a control. Thymol toxicity was evaluated in Artemia salina and in silico using Osiris® software. Differences between groups were estimated by analysis of variance, followed by Tukey post hoc test, with a 5% significance level. The results of the agar diffusion test between groups were not significantly different (P≥0.05). Thymol had potential bacteriostatic and bactericidal activity against Streptococcus mutans with respect to planktonic growth, with MIC of 100 µg/mL and MBC of 400 µg/mL. The groups GIC-0, GIC-2, and GIC-4 reduced the biofilm by approximately 10, 85, and 95%, respectively. Bacterial kinetics showed efficiency of the modified GICs for up to 96 h. GIC with thymol was effective against S. mutans, with significant inhibition of the biofilms. Analyses in silico and using Artemia salina resulted in no relevant toxicity, suggesting potential for use in humans. GIC-2 was effective against S. mutans biofilm, with decreased cell viability.

Acute body sodium depletion induces skin sodium mobilization in female Wistar rats.

NEW FINDINGS: What is the central question of this study? Can Na+ depletion mobilize Na+ from the skin reservoir in ovariectomized rats? Does oestrogen replacement change the amount and the dynamics of skin Na+ storage? Is the reduced salt appetite after Na+ depletion in ovariectomized rats with oestrogen replacement related to changes in the skin Na+ ? What is the main finding and its importance? This work demonstrated that acute body Na+ depletion induced by frusemide mobilized the osmotically inactive skin Na+ reservoir to become osmotically active. Oestrogen treatment decreased the induced Na+ intake in ovariectomized rats but did not modulate the inactive Na+ reservoir in control conditions or its mobilization induced by Na+ depletion. ABSTRACT: Oestradiol, which is an important hormone for water and electrolyte balance, also has a role in the inhibition of induced Na+ appetite. Sodium can be stored in the skin in osmotically active or inactive forms, and this skin Na+ reservoir may be involved in the control of body Na+ levels during physiopathological challenges. In this study, we investigated whether the effect of sodium depletion by frusemide can mobilize Na+ from the skin reservoir and whether oestradiol replacement changes or mobilizes the Na+ reserves in the skin. Ovariectomized Wistar rats were treated with vehicle or oestradiol for 7 days to evaluate the effects of oestrogen on the hydroelectrolyte balance, intake responses and skin Na+ and water content in basal conditions. Furthermore, the effects of oestrogen were evaluated after 24 h frusemide-induced whole-body Na+ depletion. Oestradiol-replaced rats exhibited reduced water intake without any significant changes in salt intake, Na+ excretion or water and Na+ skin content in basal conditions. After sodium depletion, both vehicle- and oestradiol-treated rats exhibited an increase in the osmotically active skin Na+ , which was associated with a decrease of the inactive skin Na+ reservoir. Oestrogen decreased the hypertonic saline intake induced by Na+ depletion, but it was not associated with any significant changes in the skin Na+ reservoir. Thus, sodium depletion is able to change the inactive-active skin Na+ reservoir balance. However, the oestrogenic modulation of sodium appetite after Na+ depletion is probably not related to the action of this hormone in the skin Na+ reservoir balance.

Immunohistochemical localization of insulin-like growth factor-1 (IGF-1) in the sheep ovary and the synergistic effect of IGF-1 and FSH on follicular development in vitro and LH receptor immunostaining.

This study analyzed IGF-1 protein immunostaining in sheep ovaries, the effect of IGF-1 alone or associated with FSH on the culture of secondary follicles, and the immunostaining of LHR protein in antral follicles before and after culture. Ovaries were collected for IGF-1 protein analysis. In experiment 1, secondary follicles were cultured in α-MEM+ (control) or α-MEM+ supplemented with IGF-1 (10, 50 or 100 ng/mL). In experiment 2, follicles were cultured in the same media of experiment 1 plus 750 ng/mL FSH. Moreover, LHR immunostaining was analyzed in fresh antral follicles and after culture in 50 ng/mL IGF-1 + FSH. The IGF-1 protein was immunolocalized in oocytes from all stages of follicle development and in the granulosa cells from secondary and antral follicles. IGF-1 did not influence (P > 0.05) follicular viability and growth (experiment 1). However, in experiment 2, 50 ng/mL IGF-1 + FSH stimulated oocyte growth (P < 0.05) and LHR immunostaining in antral follicles. Control medium, 10 or 50 ng/mL IGF-1 + FSH showed similar levels of reactive oxygen species, glutathione and active mitochondria (P > 0.05). In conclusion, the IGF-1 protein is present in all ovarian follicle stages in sheep. Moreover, the association between 50 ng/mL IGF-1 and FSH has a synergistic effect in vitro, increasing the percentage of fully grown oocytes and the intensity of immunostaining of LHR protein in oocytes and granulosa cells of cultured antral follicles.

Kaempferol promotes primordial follicle activation through the phosphatidylinositol 3-kinase/protein kinase B signaling pathway and reduces DNA fragmentation of sheep preantral follicles cultured in vitro.

The aim of this study was to evaluate the effects of kaempferol on the morphology, follicular activation, growth, and DNA fragmentation of ovine preantral follicles cultured in situ, and the effects of a phosphatidylinositol 3 kinase (PI3K) inhibitor and the expression of phosphorylated protein kinase B (pAKT) after culture. Ovine ovarian fragments were fixed for histological and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) analyses (fresh control) or cultured in α-MEM+ alone (control) or with different concentrations of kaempferol (0.1, 1, 10, or 100 µM) for 7 days. Follicles were classified as normal or atretic, primordial or growing, and the oocyte and follicle diameters were measured. Proliferating cells were analyzed and DNA fragmentation was evaluated by the TUNEL assay. Inhibition of PI3K activity was performed through pretreatment in media added with 50 µM LY294002 for 1 hr and pAKT immunohistochemistry was performed after culture in the absence or presence of LY294002. After culture, the percentage of normal follicles was similar among the treatments (p > 0.05), except for 100 µM kaempferol, which had less normal follicles (p < 0.05). Moreover, kaempferol at 10 µM showed a higher percentage of follicular activation and cell proliferation than the other treatments (p < 0.05) and a percentage of TUNEL-positive cells similar to that in the fresh control and lower than other treatments (p < 0.05). LY294002 significantly inhibited primordial follicle activation stimulated by α-MEM+ and 10 µM kaempferol and reduced pAKT expression in those follicles. In conclusion, 10 µM kaempferol promotes primordial follicle activation and cell proliferation through the PI3K/AKT pathway and reduces DNA fragmentation of ovine preantral follicles cultured in vitro.

Resveratrol has dose-dependent effects on DNA fragmentation and mitochondrial activity of ovine secondary follicles cultured in vitro.

The worldwide consumption of red wine, nuts and grapes has resulted in increased human exposure to resveratrol, which could affect reproductive function. However, the effect of resveratrol on in vitro culture of early-stage ovarian follicles has never been investigated. The aims of the present study were to evaluate the effect of resveratrol on sheep secondary follicle morphology, growth, DNA fragmentation, intracellular levels of glutathione (GSH) and active mitochondria. Secondary follicles were isolated from the ovaries and cultured for 18 days in supplemented α-MEM+ (control medium) or in control medium containing resveratrol (2, 10 or 30 µM). The parameters analyzed were morphology, antrum formation, follicle diameter, DNA fragmentation, GSH levels and mitochondrial activity. After 18 days, all resveratrol groups significantly decreased the percentages of morphologically normal follicles compared with the control group (α-MEM+). Antrum formation was higher in both α-MEM+ and 2 µM resveratrol groups than in the 10 µM resveratrol group. In addition, 30 µM resveratrol increased the percentage of oocytes with DNA damage compared with the control. Oocytes from follicles treated with 10 or 30 µM resveratrol significantly decreased intracellular GSH levels compared with the 2 µM resveratrol group. Moreover, follicles in α-MEM+ (control) showed more active mitochondria than those in 10 or 30 µM resveratrol. In conclusion, ovine isolated secondary follicles are able to grow to the antral stage after in vitro culture in medium containing 2 µM resveratrol, maintaining the same rates of DNA damage, GSH levels and mitochondrial function as the control medium. However, the addition of 30 µM resveratrol increased DNA fragmentation and oxidative stress through decreasing mitochondrial activity.

Use of protocatechuic acid as the sole antioxidant in the base medium for in vitro culture of ovine isolated secondary follicles.

This study evaluated the effect of the protocatechuic acid (PCA) as the sole antioxidant in the base medium for in vitro culture of ovine secondary follicles. Secondary follicles (200-230 µm) were isolated and cultured in α-minimal essential medium supplemented with BSA, insulin, glutamine and hypoxanthine (α-MEM: antioxidant-free medium) or α-MEM also added by transferrin, selenium and ascorbic acid (α-MEM+: with antioxidant) or α-MEM added by PCA (56.25; 112.5; 225; 450; or 900 µg/ml). Moreover, after culture, oocytes were matured and the chromatin configuration and DNA fragmentation were evaluated. After 12 days, the treatment containing 56.25 µg/ml PCA showed higher percentage of normal follicles than control medium or the other treatments (p < .05), except for 900 µg/ml PCA (p > .05). The antrum formation was significantly higher in treatments containing 56.25, 112.5 or 900 µg/ml PCA, compared to the α-MEM and similar (p > .05) to the other treatments. The rates of fully grown oocytes (≥110 µm) were similar (p > .05) among all treatments containing PCA and α-MEM+, and those were superior (p < .05) than α-MEM, except for 450 µg/ml PCA (p > .05). GSH levels and mitochondrial activity were higher (p < .05) in α-MEM+ than in α-MEM and similar (p > .05) to all PCA treatments. The rates of meiotic resumption and DNA fragmentation were similar (p > .05) among α-MEM+ and 56.25 µg/ml PCA. In conclusion, PCA at 56.25 µg/ml as the sole antioxidant added to the medium for ovine isolated secondary follicle culture maintains follicular survival, GSH and active mitochondria levels, meiotic developmental competence and DNA integrity of cultured oocytes.

Expression of TNF-α system members in bovine ovarian follicles and the effects of TNF-α or dexamethasone on preantral follicle survival, development and ultrastructure in vitro.

This study was conducted to detect the protein expression of TNF-α system members (TNF-α/TNFR1/TNFR2) in bovine ovarian follicles and to evaluate the effects of TNF-α or dexamethasone on the survival and growth of primordial follicles in vitro, as well as on gene expression in cultured ovarian tissue. It was hypothesized that TNF-α induces follicular atresia in ovarian tissues cultured in vitro, and that dexamethasone suppresses the production of endogenous TNF-α, which can improve follicle viability in vitro. Ovarian fragments were cultured for 6days in α-MEM+ supplemented with TNF-α (0, 1, 10, 100 or 200ng/ml) or dexamethasone (0, 1, 10, 100 or 200ng/ml). After culture, the expression of mRNAs for BCL-2, BAX, P53, TNF-α, and CASP3 and CASP6 were evaluated. Immunohistochemical results showed that the TNF-α system members, were detected in bovine preantral and antral follicles. After 6days, the TNF-α (10ng/ml) treatment reduced the percentage of normal preantral follicles and increased the number of TUNEL-positive cells in cultured tissue. Dexamethasone (10ng/ml) during 6days of culture did maintain the percentage of normal follicles and the ultrastructure of follicles, while the presence of TNF-α or dexamethasone did not influence primordial follicle activation. However, TNF-α or dexamethasone had no effect on the levels of mRNA for P53, BCL-2, BAX and CASP6, in cultured tissues, but the presence of dexamethasone reduced the levels of CASP3 compared to ovarian slices cultured in control medium (α-MEM+). In conclusion, proteins of the TNF-α system are expressed at different bovine follicle stages. The addition of TNF-α in culture reduces follicle survival and increases the number of apoptotic cells in ovarian tissue, while the presence of dexamethasone maintains follicle ultrastructure in cultured tissue.

Do restinga cnemidophorine lizards run on empty along the Brazilian coast?

Abstract The number of individuals with empty stomachs in a population (i.e. the proportion of individuals running on empty) can be used as a simple index of instantaneous energy balance of some organisms such as lizards and fishes. In this study, we aimed to analyze the proportion of empty stomachs in 16 populations of five cnemidophorine species (Glaucomastix abaetensis, Ameivula ocellifera, Glaucomastix littoralis, Contomastix lacertoides and Ameivula nativo) along approximately 5,000 km the Brazilian coast. A total of 550 individuals had their stomach contents removed and identified in the laboratory. Our results showed that the proportion of individuals running on empty varied from 0 to 11.1% among the different populations and species. These proportions are suggestive that the five studied species would be in an overall positive energy balance at the time of the study.

Resumo O número de indivíduos com estômagos vazios em uma população (i.e. a proporção de indivíduos correndo no vazio) pode ser usado como um índice simples do balanço energético instantâneo de organismos como lagartos e peixes. Este estudo teve como objetivo analisar a proporção de estômagos vazios em 16 populações pertencentes a cinco espécies de cnemidophorines (Glaucomastix abaetensis, Ameivula ocellifera, Glaucomastix littoralis, Contomastix lacertoides e Ameivula nativo) ao longo de cerca de 5,000 km da costa leste do Brasil. Um total de 550 indivíduos tiveram seus conteúdos estomacais removidos e analisados em laboratório. Nossos resultados mostraram que a proporção de indivíduos correndo no vazio variou entre 0 a 11.1% entre as diferentes populações e espécies. Essas proporções sugerem que as cinco espécies estudadas estariam em geral em um balanço energético positivo, quando da realização deste estudo.

A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro

Abstract Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called restinga and campo nativo habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.

Resumo Embora atualmente exista um conjunto de estudos sobre aspectos ecológicos de algumas espécies de répteis e de anfíbios que ocorrem nas planícies costeiras arenosas brasileiras (incluindo os chamados habitats de restinga e de campo nativo), há relativamente poucas informações sobre a composição de espécies geralmente associada a esses ambientes. Durante 31 anos (1988-2019) de estudos herpetológicos realizados em restingas por nossa equipe de pesquisa do Laboratório de Ecologia de Vertebrados (Departamento de Ecologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil) nós estudamos comunidades de répteis e de anfíbios e realizamos diferentes estudos com métodos semelhantes em 70 localidades de dez diferentes Estados ao longo da costa brasileira. Nossas pesquisas resultaram em registros de 87 espécies de répteis (cinco tartarugas, dois crocodilianos, seis anfisbênios, 36 lagartos e 39 serpentes) de 24 famílias, e 77 espécies de anfíbios anuros de nove famílias. Estudamos vários tópicos de história natural sobre anuros e répteis, que resultaram na publicação de alguns estudos ecológicos específicos, especialmente em relação a algumas espécies, abrangendo ecologia populacional e de comunidades, forrageamento e dieta, horário de atividade de espécies, termorregulação, reprodução, uso do microhabitat e parasitismo por ecto e endoparasitas. Nossos resultados ao longo dessas três décadas também contribuíram para a descrição de quatro novas espécies de lagartos (Ameivula nativo, Glaucomastix littoralis, G. abaetensis e G. itabaianensis). Nossos estudos constituem uma importante contribuição para o conhecimento da ecologia de répteis e de anfíbios anuros nesses ecossistemas, bem como para a conservação dos ecossistemas de restinga. A lista de espécies apresentada neste estudo, com base em nossos registros de herpetofauna das planícies costeiras arenosas, fornece para muitas localidades ao longo da costa brasileira o conhecimento necessário sobre a ocorrência de espécies, incluindo a presença de espécies endêmicas e/ ou ameaçadas de extinção, que podem ser úteis para muitas ações de conservação.

Supplemented base medium containing Amburana cearensis associated with FSH improves in vitro development of isolated goat preantral follicles.

The effects of Amburana cearensis ethanolic extract, with or without addition of a mix of supplements associated or not with FSH, on in vitro morphology and development of caprine secondary follicles were evaluated. In experiment 1, isolated follicles (250 µm in diameter) were cultured for 12 days in alpha-modified minimal essential medium (α-MEM) alone (control) or in medium composed of different concentrations of A. cearensis extract (Amb 0.1; 0.2, or 0.4 mg/mL). In experiment 2, culture media were α-MEM or Amb 0.2 mg/mL (both without supplements), or these same media supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred as α-MEM(+) and Amb 0.2(+), respectively), or these last groups also supplemented with sequential FSH (100 ng/mL from Day 0 to Day 6; 500 ng/mL from Day 6 to Day 12), constituting groups α-MEM(+) + FSH and Amb 0.2(+) + FSH. At the end of culture in experiment 1, control medium (α-MEM) and Amb 0.2 mg/mL had higher percentages (P < 0.05) of morphologically normal follicles and percentage of fully grown oocytes, i.e., oocyte greater than 110 µm, compared to the other A. cearensis extract concentrations. In experiment 2, all supplemented media had higher percentages (P < 0.05) of normal follicles and antrum formation than nonsupplemented media. In addition, follicles cultured in Amb 0.2(+) + FSH showed an average increase in diameter higher (P < 0.05) than the other treatments. Oocytes cultured in both treatments supplemented with FSH showed greater glutathione and active mitochondria levels than nonsupplemented media but similar to the other treatments. In conclusion, A. cearensis extract (0.2 mg/mL) added by supplements and FSH improves follicular growth. Therefore, it can be an alternative culture medium for goat preantral follicle development.

Effects of a fire on a population of treefrogs (Scinax cf. alter, Lutz) in a restinga habitat in southern Brazil / Efeitos de uma queimada em uma população de pererecas (Scinax cf. alter) em um habitat de restinga no sul do Brasil

The area of the Dunas da Joaquina, in Santa Catarina island, contains one of the most important remnants of restinga habitat in Santa Catarina State, southern Brazil. In December 2003, a fire occurred in a portion of this area, affecting most of the vegetation, including the bromeliad community. In this study, the density of individuals and the diet composition of the bromelicolous treefrog Scinax cf. alter were compared between the area affected by the fire and an adjacent unburned area. One-hundred-and-fifty-eight ground bromeliads (Vriesea friburguensis) were dissected and searched for the presence of treefrogs among their leaves. We found 30 frogs in 29.5 percent (23/78) of the bromeliads from the unburned site, with a mean of 1.3 frogs per rosette, and 15 frogs in 12.5 percent (10/80) of the bromeliads from the burned site, with a mean of 1.6 frogs per rosette. Eight (27 percent) of the frogs from the unburned site and eleven (73 percent) of those from the burned site had empty stomachs. Frogs from the burned site also contained less prey per stomach than those from the unburned site. The data suggest that the fire has negatively affected the local population of Scinax cf. alter, though it is possible that the population can recover.

A área das Dunas da Joaquina, na Ilha de Santa Catarina, contém um dos remanescentes mais importantes de habitat de restinga no Estado de Santa Catarina, no Sul do Brasil. Em dezembro de 2003, ocorreu uma queimada em uma porção desta área, afetando a maior parte da vegetação, incluindo a comunidade de bromélias. Neste estudo, comparamos a densidade de indivíduos e a composição da dieta da perereca bromelícola Scinax cf. alter entre a área afetada pelo fogo e a uma área adjacente que não foi afetada. Cento e cinqüenta e oito bromélias de chão (Vriesea friburguensis) foram dissecadas e examinadas quanto à presença de pererecas entre suas folhas. Foram encontradas 30 pererecas em 29,5 por cento (23/78) das bromélias da área não-queimada, com uma média de 1,3 indivíduos por roseta; e 15 pererecas em 12,5 por cento (10/80) das bromélias da área queimada, com uma média de 1,6 indivíduos por roseta. Oito (27 por cento) dos animais da área não-queimada; e onze (73 por cento) dos da área queimada estavam com o estômago vazio. As pererecas da área queimada também tiveram menos presas por estômago que as da área não-queimada. Os dados sugerem que o fogo afetou negativamente a população local de Scinax cf. alter, embora seja possível a sua recuperação.

Development of shock wave assisted therapeutic devices and establishment of shock wave therapy.

In order to exploit systems for shock wave therapy, we are working for the development of clinical devices that are based on the concept of shock waves or related phenomena. The paper describes these new therapeutic devices designed for the minimally invasive approach to vascular thromboloysis, selective dissection of tissues, and drug or DNA delivery. To investigate the response of cells to shock loading, a precise method of shock waves generation in space and time has been developed. This method has been studied for application in cardiovascular therapy, cancer treatment, and cranioplasty in close vicinity of the brain. A laser ablation shock wave assisted particle acceleration device has been developed for delivering drug and DNA into soft targets in the human body. The penetration depth of microparticles observed in the experimental targets is believed to be sufficient for pharmacological treatments. In order to achieve an efficient method for rapid revascularization of cerebral thrombosis, a laser induced liquid jet (LILJ) system has been developed. The LILJ has been successfully applied for selective dissection of soft tissue preserving nerve and blood vessels. The system has been further improved by using piezoelectric actuators to drive the liquid jets, as an alternative to pulse laser.

Diet and foraging of the endemic lizard Cnemidophorus littoralis (Squamata, Teiidae) in the restinga de Jurubatiba, Macaé, RJ / Dieta e forrageamento do lagarto endêmico Cnemidophorus littoralis (Squamata, Teiidae) da restinga de Jurubatiba, Macaé - RJ

We investigated the diet and foraging of the endemic teiid lizard Cnemidophorus littoralis in a restinga habitat in Jurubatiba, Macaé - RJ. The stomach contents were removed, analyzed and identified to the Order level. There was no relationship between C. littoralis morphological variables and number, length or volume of preys. Termites (48.7 percent) and larvae (35.5 percent) were the most important prey items which occurred in the examined lizards´ stomachs. The diet did not differ between males and females. Cnemidophorus littoralis is an active forager and predominantly consumes relatively sedentary prey or prey that is aggregated in the environment. We also found an intact and undigested hatchling of the crepuscular/nocturnal gekkonid lizard Hemidactylus mabouia in the stomach of an adult male of C. littoralis, which indicates that C. littoralis is a potential source of mortality for individuals of H. mabouia in the restinga de Jurubatiba.

Investigou-se a dieta e o forrageamento do lagarto endêmico Cnemidophorus littoralis na restinga de Jurubatiba, Macaé - RJ. Os conteúdos estomacais foram removidos, analisados e identificados no nível de Ordem. Não houve relação entre as variáveis morfológicas de C. littoralis e o número, tamanho e volume das presas. Cupins (48,7 por cento) e larvas (35,5 por cento) foram os itens de presas mais importantes nos estômagos dos lagartos examinados. A dieta não diferiu entre os machos e as fêmeas. Cnemidophorus. littoralis é um lagarto forrageador ativo e consome, predominantemente, presas relativamente sedentárias ou que ocorrem agregadas no ambiente. Foi encontrado também no estômago de um macho adulto de C. littoralis um jovem lagarto gekkonídeo crepuscular/noturno Hemidactylus mabouia intacto e não digerido, o que indica C. littoralis como uma fonte de mortalidade para indivíduos de H. mabouia na restinga de Jurubatiba.

Drugs for treating paracoccidioidomycosis.

BACKGROUND: Paracoccidioidomycosis is a fungal infection found in particular geographic localities in Latin America. Treatment can last for up to two years is often associated with complications, including relapse, but people may die without it. OBJECTIVES: To evaluate drugs used for treating paracoccidioidomycosis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group Specialized Register (January 2006), CENTRAL (The Cochrane Library 2005, Issue 4), PubMed (1966 to January 2006), EMBASE (1974 to January 2006), LILACS (1982 to January 2006), conference proceedings, and reference lists. We also contacted researchers and pharmaceutical companies. SELECTION CRITERIA: Randomized controlled trials comparing drugs for treating people with paracoccidioidomycosis. DATA COLLECTION AND ANALYSIS: Two authors independently assessed trial eligibility and methodological quality, and extracted data, including adverse events. MAIN RESULTS: One trial with 42 participants met the inclusion criteria that compared imidazoles (itraconazole and ketoconazole) with sulfadiazine. No difference was detected for cure or clinical improvement, or serological titres after 10 months of treatment, and there was no difference detected in adverse events. AUTHORS' CONCLUSIONS: The small number of participants and the short follow-up period impede definitive conclusions.