Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone.

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Effects of antifreeze protein from Lolium perenne L. (LpAFP) in the vitrification of in vitro-produced bovine embryos.

In the present study, the cryoprotective effects of Lolium perenne antifreeze protein (LpAFP) on the vitrification of bovine embryos were evaluated. In vitro-produced blastocysts were divided into two groups: the control group (CG) without the addition of LpAFP and the treatment group (TG) with the addition of 500 ng/ml of LpAFP in the equilibrium and vitrification solution. Vitrification was carried out by transferring the blastocysts to the equilibrium solution [7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO)] for 2 min and then to the vitrification solution (15% EG, 15% DMSO and 0.5M sucrose). The blastocysts were deposited on a cryotop device and submerged in liquid nitrogen. Warming was carried out in three steps in solutions with different sucrose concentrations (1.0, 0.5, and 0.0 M, respectively). Embryos were evaluated for re-expansion/hatching, the total cell count, and ultrastructural analysis. There was no significant difference in the re-expansion rate 24 h after warming; however, there was variation (P < 0.05) in the hatching rate in the TG and the total number of cells 24 h after warming was higher in the TG (114.87 ± 7.24) when compared with the CG (91.81 ± 4.94). The ultrastructural analysis showed changes in organelles related to the vitrification process but, in the TG, there was less damage to mitochondria and rough endoplasmic reticulum compared with the CG. In conclusion, the addition of 500 ng/ml of LpAFP during the vitrification of in vitro-produced bovine embryos improved the hatching rate and total cell number of blastocysts after warming and mitigated intracellular damage.

Manejo anestésico perioperatorio de la deficiencia de acil-coenzima A deshidrogenasa de cadena muy larga / Perioperative anesthetic management of very long-chain acyl-coenzyme a dehydrogenase deficiency

La deficiencia de acil-coenzima A deshidrogenasa de cadena muy larga (VLCADD) es un trastorno infrecuente del metabolismo de β-oxidación de los ácidos grasos que origina susceptibilidad a hipoglucemia, fallo hepático, cardiomiopatía y rabdomiólisis durante las situaciones catabólicas. Reportamos el caso de un varón de 10 años de edad programado para la colocación de catéter venoso central totalmente implantado durante su hospitalización por rabdomiólisis, que fue exitosamente gestionada con anestesia general con óxido nitroso, sevoflurano y remifentanilo. No se produjo hipoglucemia y los niveles de creatina quinasa no se incrementaron durante el periodo perioperatorio. Describimos las dificultades a que nos enfrentamos, y las estrategias utilizadas para evitar mayor descompensación de la enfermedad debida al estrés quirúrgico.(AU)

Very long-chain acyl-coenzyme A dehydrogenase deficiency (VLCADD) is a rare disorder of β-oxidation fatty acid metabolism that results in susceptibility to hypoglycemia, liver failure, cardiomyopathy and rhabdomyolysis during catabolic situations. We report the case of a 10-year-old male undergoing a totally implanted central venous catheter placement during hospitalization for rhabdomyolysis, who was successfully managed with general anesthesia with nitrous oxide, sevoflurane and remifentanil. No hypoglycemia occurred and creatine kinase levels did not increase in the perioperative period. We describe the challenges encountered and the strategies used to avoid further decompensation of the disease due to surgical stress.(AU)

Perioperative anesthetic management of very long-chain acyl-coenzyme a dehydrogenase deficiency.

Very long-chain acyl-coenzyme A dehydrogenase deficiency is a rare disorder of ß-oxidation fatty acid metabolism that results in susceptibility to hypoglycemia, liver failure, cardiomyopathy and rhabdomyolysis during catabolic situations. We report the case of a 10-year-old male undergoing a totally implanted central venous catheter placement during hospitalisation for rhabdomyolysis, who was successfully managed with general anesthesia with nitrous oxide, sevoflurane and remifentanil. No hypoglycemia occurred and creatine kinase levels did not increase in the perioperative period. We describe the challenges encountered and the strategies used to avoid further decompensation of the disease due to surgical stress.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Comparación del bloqueo del grupo de nervios pericapsulares (PENG) y la analgesia epidural después de una artroplastia total de cadera: análisis retrospectivo / Comparison of pericapsular nerve group (PENG) block and epidural analgesia following total hip arthroplasty: a retrospective analysis

Introducción y objetivos: La artroplastia total de cadera (THA) es un procedimiento traumatológico cada vez más común, que comporta dolor postoperatorio de moderado a severo. El bloqueo del grupo de nervios pericapsulares (PENG) es un bloqueo reciente que parece aportar analgesia adecuada sin bloqueo motor significativo. El objeto de este estudio es comparar retrospectivamente la eficacia analgésica y la seguridad del bloqueo PENG y la analgesia epidural, en pacientes sometidos a THA. Material y métodos: Se trata de un estudio retrospectivo observacional durante un periodo de un año de los pacientes sometidos a THA primaria, utilizando analgesia epidural o bloqueo PENG ecoguiado con inyección única. Los datos relativos a las características demográficas, técnicas quirúrgicas y anestésicas, puntuaciones de dolor, consumo de opioides, complicaciones y tiempo hasta el alta hospitalaria se recopilaron de los registros institucionales, comparándose entre los dos grupos (analgesia epidural vs. bloqueo PENG). Resultados: No se encontraron diferencias significativas en cuanto a las puntuaciones de dolor, el consumo de opioides y el tiempo medio hasta el alta hospitalaria entre los dos grupos. Las puntuaciones de dolor en reposo (1,2 epidural vs. 1,67 PENG) o con movimiento (3,95 epidural vs. 3,72 PENG) fueron similares entre ambos grupos. El número total de complicaciones fue superior en el grupo de analgesia epidural (50% epidural vs. 5% PENG). Se reportó parestesia en ambos grupos. Solo se reportaron bloqueo motor, sedación, náuseas y complicaciones relacionadas con el catéter en el grupo de analgesia epidural. Conclusiones: El bloqueo PENG parece ser equivalente a la analgesia epidural en cuanto a calidad de la analgesia postoperatoria para los pacientes sometidos a THA primaria, lo cual respalda el uso rutinario de este bloqueo en estos pacientes. La baja tasa de complicaciones reportadas limita las conclusiones sobre esta cuestión.(AU)

Introduction and objectives: Total hip arthroplasty (THA) is an increasingly common orthopedic pro-cedure, with moderate to severe postoperative pain. Pericapsular nerve group (PENG) block is a recent block that seems to provide adequate analgesia without significant motor blockade. The aim of this study is to retrospectively compare the analgesic efficacy and safety of PENG block with those of epidural analgesia, in patients undergoing THA. Material and methods: This is a retrospective observational study of patients who underwent primary THA, submitted to epidural analgesia or single-shot ultrasound-guided PENG block, during a one-year period. Data regarding demographic characteristics, surgery and anesthesia techniques, pain scores, opioid consumption, complications and time to hospital discharge were retrieved from institutional records and compared between the 2 groups (epidural analgesia vs PENG block). Results: No significant difference was found regarding pain scores, opioid consumption, and mean time to hospital discharge between the 2 groups. Pain scores at rest (1.20 epidural vs 1.67 PENG) or with movement (3.95 epidural vs 3.72 PENG) were similar between groups. Total number of complications was higher in the epidural analgesia group (50% epidural vs 5% PENG). Paresthesia was reported in both groups. Motor block, sedation, nausea and catheter-related complications were only found in the epidural analgesia group. Conclusions: PENG block seems to be equivalent to epidural analgesia regarding quality of postop-erative analgesia for patients subject to primary THA, supporting routine use of this block in these patients. The low rate of reported complications limits conclusions on this topic.(AU)

Comparison of pericapsular nerve group (PENG) block and epidural analgesia following total hip arthroplasty: A retrospective analysis.

INTRODUCTION AND OBJECTIVES: Total hip arthroplasty (THA) is an increasingly common orthopaedic procedure, with moderate to severe postoperative pain. Pericapsular nerve group (PENG) block is a recent block that seems to provide adequate analgesia without significant motor blockade. The aim of this study is to retrospectively compare the analgesic efficacy and safety of PENG block with those of epidural analgesia, in patients undergoing THA. MATERIAL AND METHODS: This is a retrospective observational study of patients who underwent primary THA, submitted to epidural analgesia or single-shot ultrasound-guided PENG block, during a one-year period. Data regarding demographic characteristics, surgery and anaesthesia techniques, pain scores, opioid consumption, complications and time to hospital discharge were retrieved from institutional records and compared between the 2 groups (epidural analgesia vs PENG block). RESULTS: No significant difference was found regarding pain scores, opioid consumption, and mean time to hospital discharge between the 2 groups. Pain scores at rest (1.20 epidural vs 1.67 PENG) or with movement (3.95 epidural vs 3.72 PENG) were similar between groups. Total number of complications was higher in the epidural analgesia group (50 % epidural vs 5% PENG). Paresthesia was reported in both groups. Motor block, sedation, nausea and catheter-related complications were only found in the epidural analgesia group. CONCLUSIONS: PENG block seems to be equivalent to epidural analgesia regarding quality of postoperative analgesia for patients subject to primary THA, supporting routine use of this block in these patients. The low rate of reported complications limits conclusions on this topic.

Physical and rehabilitation medicine intervention in stroke in the acute hospital setting: A cross-sectional study / Intervención de la medicina física y de rehabilitación en el accidente cerebrovascular en el ámbito hospitalario agudo: un estudio transversal

Introduction: Stroke represents the main cause of death and disability in Portugal. Resulting functional deficits are widely recognized. This work aims to evaluate the variation in functionality of stroke patients in the acute hospital setting under a rehabilitation program. Material and methods: Cross-sectional study of patients admitted to the Neurology department, from January to June 2019, with acute stroke. The variation in functionality was assessed using the Barthel index. Statistical analysis used Student's t-test and Spearman's correlation coefficient, with a p-value≤0.05 as significant. Results: 106 patients with mean age of 63.7±14.2 years and a male predominance (60.4%) were included. Patients started rehabilitation program at 1.37±1.19 days after admission. A gain in functionality between admission and discharge was identified (50.18±32.37 versus 68.73±28.94, p<0.001). A significantly greater increase was observed in patients diagnosed under code stroke protocol (CSP) (p=0.021) and undergoing some type of acute phase treatment (p=0.017). From 90.5% of the patients that pursued rehabilitation after discharge, 40.6% were referred to an inpatient unit on average 12.7±7.0 days after admission. Discussion: In this study, Physical and Rehabilitation Medicine (PRM) provided early rehabilitation care to stroke patients. According to international evidence this is associated with greater functional gains. The variation in functionality verified during hospitalization demonstrates the importance of PRM in the acute hospital, assessing the rehabilitation needs after hospital discharge and maximizing outpatient rehabilitation. Diagnosis under CSP and undergoing acute treatment were determinants of greater functional improvement. Conclusion: PRM plays a central role in the early management of functional impairment resulting from stroke and in the post-discharge guidance of patients.(AU)

Introducción: El accidente cerebrovascular representa la principal causa de muerte y incapacidad en Portugal. Los déficits funcionales resultantes son ampliamente reconocidos. Este trabajo tiene como objetivo evaluar la variación en la funcionalidad de los pacientes con accidente cerebrovascular en el entorno hospitalario agudo bajo un programa de rehabilitación. Material y métodos: Estudio transversal de pacientes ingresados en el servicio de neurología, de enero a junio de 2019, con accidente cerebrovascular agudo. La variación en la funcionalidad se evaluó mediante el índice de Barthel. El análisis estadístico utilizó la prueba «t» de Student y el coeficiente de correlación de Spearman, con un valor p≤0,05 como significativo. Resultados: Se incluyeron 106 pacientes con edad media de 63,7±14,2 años y predominio del sexo masculino (60,4%). Los pacientes iniciaron el programa de rehabilitación a los 1,37±1,19 días después del ingreso. Se identificó una ganancia de funcionalidad entre el ingreso y el alta (50,18±32,37 versus 68,73±28,94, p<0,001). Se observó un aumento significativamente mayor en los pacientes diagnosticados bajo el protocolo de ictus (p=0,021) y sometidos a algún tipo de tratamiento en fase aguda (p=0,017). Del 90,5% de los pacientes que continuaron rehabilitación tras el alta, el 40,6% fueron derivados a una unidad de rehabilitación, en promedio 12,7±7,0 días después del ingreso. Discusión: En este estudio la medicina física y de rehabilitación (MFR) proporcionó atención de rehabilitación temprana a pacientes con accidente cerebrovascular. Según la evidencia internacional esto se asocia con mayores ganancias funcionales. La variación en la funcionalidad verificada durante la hospitalización demuestra la importancia de la MFR en el hospital de agudos, evaluando las necesidades de rehabilitación después del alta hospitalaria y maximizando la rehabilitación ambulatoria.(AU)

Physical and rehabilitation medicine intervention in stroke in the acute hospital setting: A cross-sectional study.

INTRODUCTION: Stroke represents the main cause of death and disability in Portugal. Resulting functional deficits are widely recognized. This work aims to evaluate the variation in functionality of stroke patients in the acute hospital setting under a rehabilitation program. MATERIAL AND METHODS: Cross-sectional study of patients admitted to the Neurology department, from January to June 2019, with acute stroke. The variation in functionality was assessed using the Barthel index. Statistical analysis used Student's t-test and Spearman's correlation coefficient, with a p-value≤0.05 as significant. RESULTS: 106 patients with mean age of 63.7±14.2 years and a male predominance (60.4%) were included. Patients started rehabilitation program at 1.37±1.19 days after admission. A gain in functionality between admission and discharge was identified (50.18±32.37 versus 68.73±28.94, p<0.001). A significantly greater increase was observed in patients diagnosed under code stroke protocol (CSP) (p=0.021) and undergoing some type of acute phase treatment (p=0.017). From 90.5% of the patients that pursued rehabilitation after discharge, 40.6% were referred to an inpatient unit on average 12.7±7.0 days after admission. DISCUSSION: In this study, Physical and Rehabilitation Medicine (PRM) provided early rehabilitation care to stroke patients. According to international evidence this is associated with greater functional gains. The variation in functionality verified during hospitalization demonstrates the importance of PRM in the acute hospital, assessing the rehabilitation needs after hospital discharge and maximizing outpatient rehabilitation. Diagnosis under CSP and undergoing acute treatment were determinants of greater functional improvement. CONCLUSION: PRM plays a central role in the early management of functional impairment resulting from stroke and in the post-discharge guidance of patients.

Species diversity, novel interactions and absence of well-supported host-guided phylogenetic groupings of Neotropical Alternaria isolates causing foliar lesions in Solanaceae.

AIM: To report the characterization of 120 Alternaria isolates inducing early blight-like foliar lesions in nine species of five Solanaceae genera collected across all macrogeographical Brazilian regions. MATERIAL AND RESULTS: Phylogenetic relationships were assessed via analyses of the Alternaria alternata allergenic protein-coding, glyceraldehyde-3-phosphate dehydrogenase and the calmodulin gene sequences. Most of the tomato isolates were placed into the Alternaria linariae cluster, whereas most of the potato isolates were grouped with Alternaria grandis. Novel host-pathogen interactions were also reported. Seventeen isolates were selected for morphometrical characterization, and a subsample of 13 isolates was employed in pathogenicity assays on tomato, potato, eggplant, scarlet eggplant, Capsicum annuum, Datura stramonium, Physalis angulata and Nicotiana tabacum. Eleven isolates were able to induce foliar lesions in tomatoes but none in C. annuum. Potato was susceptible to a subgroup of isolates but displayed a subset of isolate-specific interactions. Morphological traits were in overall agreement with molecular and host range data. CONCLUSION: Alternaria linariae and A. grandis were confirmed as the major causal agents of tomato and potato early blight, respectively. However other Alternaria species are also involved with early blight in solanaceous hosts in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: The diversity and host-specific patterns of the Alternaria isolates from Solanaceae may have practical implications in establishing effective early blight genetic resistance and cultural management strategies especially for tomato and potato crops.

Isolates of Bacillus thuringiensis from Maranhão biomes with potential insecticidal action against Aedes aegypti larvae (Diptera, Culicidae) / Isolados de Bacillus thuringiensis de biomas maranhenses com potencial ação inseticida contra larvas de Aedes aegypti (Diptera, Culicidae)

Abstract Entomopathogenic agents are viable and effective options due to their selective action against insects but benign effects on humans and the environment. The most promising entomopathogens include subspecies of Bacillus thuringiensis (Bt), which are widely used for the biological control of insects, including mosquito vectors of human pathogens. The efficacy of B. thuringiensis toxicity has led to the search for new potentially toxic isolates in different regions of the world. Therefore, soil samples from the Amazon, Cerrado and Caatinga biomes of the state of Maranhão were evaluated for their potential larvicidal action against Aedes aegypti. The isolates with high toxicity to mosquito larvae, as detected by bioassays, were subjected to histological evaluation under a light microscope to identify the genes potentially responsible for the toxicity. Additionally, the toxic effects of these isolates on the intestinal epithelium were assessed. In the new B. thuringiensis isolates toxic to A. aegypti larvae, cry and cyt genes were amplified at different frequencies, with cry4, cyt1, cry32, cry10 and cry11 being the most frequent (33-55%) among those investigated. These genes encode specific proteins toxic to dipterans and may explain the severe morphological changes in the intestine of A. aegypti larvae caused by the toxins of the isolates.

Resumo Os agentes entomopatógenos são alternativas viáveis e eficazes, devido à sua ação seletiva para insetos sendo inofensivos ao homem e ao meio ambiente. Dentre os entomopatógenos mais promissores, destacam-se as subespécies de Bacillus thuringiensis (Bt) amplamente utilizadas no controle biológico de insetos incluindo espécies de mosquitos vetores de agentes patogênicos ao homem. A eficiência da toxicidade de Bt incentiva a prospecção de novos isolados em diversas regiões do mundo. Desta forma, em busca de novos isolados de B. thuringiensis potencialmente tóxicos, amostras de solo provenientes dos biomas Amazônia, Cerrado e Caatinga do estado do Maranhão foram avaliadas em relação ao seu potencial larvicida para Aedes aegypti. Os isolados que provocaram elevada toxicidade para larvas do mosquito, detectada por bioensaios, foram avaliados em relação aos potenciais genes responsáveis pela atividade tóxica, além da avaliação de efeitos tóxicos no epitélio intestinal através de análises histológicas em microscopia de luz. Os novos isolados de Bt tóxicos para larva de A. aegypti amplificaram frequências diferentes de genes cry e cyt sendo os mais frequentes (55-33%) os cry4, cyt1, cry32, cry10 e cry11 dentre os investigados. Esses genes codificam para proteínas tóxicas específicas para ordem Diptera, e podem explicar as severas alterações morfológicas provocadas pelas toxinas dos isolados observadas no intestino das larvas de A. aegypti.

Isolates of Bacillus thuringiensis from Maranhão biomes with potential insecticidal action against Aedes aegypti larvae (Diptera, Culicidae).

Entomopathogenic agents are viable and effective options due to their selective action against insects but benign effects on humans and the environment. The most promising entomopathogens include subspecies of Bacillus thuringiensis (Bt), which are widely used for the biological control of insects, including mosquito vectors of human pathogens. The efficacy of B. thuringiensis toxicity has led to the search for new potentially toxic isolates in different regions of the world. Therefore, soil samples from the Amazon, Cerrado and Caatinga biomes of the state of Maranhão were evaluated for their potential larvicidal action against Aedes aegypti. The isolates with high toxicity to mosquito larvae, as detected by bioassays, were subjected to histological evaluation under a light microscope to identify the genes potentially responsible for the toxicity. Additionally, the toxic effects of these isolates on the intestinal epithelium were assessed. In the new B. thuringiensis isolates toxic to A. aegypti larvae, cry and cyt genes were amplified at different frequencies, with cry4, cyt1, cry32, cry10 and cry11 being the most frequent (33-55%) among those investigated. These genes encode specific proteins toxic to dipterans and may explain the severe morphological changes in the intestine of A. aegypti larvae caused by the toxins of the isolates.

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro.

This study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150-200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal-Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro.

The aims of this study were to investigate the effects of epidermal growth factor (EGF) and progesterone on the development, viability and the gene expression of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (∼0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5 °C, with 5% CO2 in air, for 18 days, in TCM-199+ (n = 63) alone (control medium) or supplemented with 10 ng/mL progesterone (n = 64), 10 ng/mL EGF (n = 61) or both EGF and progesterone (n = 66). The effects of these treatments on growth, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and cyclin B1 were evaluated, significantly different (p < 0.05). The results showed that there was a progressive increase in follicular diameter in all treatments, but only follicles cultured in medium supplemented with EGF had increased significantly in diameter when compared to follicles cultured in the control medium at the end of the culture period, significantly different (p < 0.05). A positive interaction between EGF and progesterone was not observed. In addition, the presence of EGF, progesterone or both in culture medium did not influence the rate of follicle survival and antrum formation. However, the presence of only progesterone in cultured medium increased the expression of mRNAs for GDF9 and cyclin B1 in oocytes. EGF also significantly increased the levels of mRNAs for cMOS and GDF9 when compared to follicles cultured in control medium. Ultrastructural analyzes showed that cultured follicles in all treatments maintained the integrity of granulosa cells. In conclusion, the EGF promotes the development of secondary follicles cultured in vitro for 18 days and increases the expression of cMOS and GDF9, while progesterone alone or in association with EGF have not a positive effect on follicular growth. However, progesterone increases the expression of GDF9 and cyclin B1 in 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.

mRNA expression profile of the TNF-α system in LH-induced bovine preovulatory follicles and effects of TNF-α on gene expression, ultrastructure and expansion of cumulus-oocyte complexes cultured in vitro.

This study evaluated (1) the effects of in vivo GnRH treatment on mRNA expression of TNF-α system (TNF-α, TNFR1 and TNFR2) in granulosa cells of bovine preovulatory follicles, (2) the in vitro influence of gonadotropins on mRNA expression of TNF-α system in cultured cumulus cells, (3) the protein expression of the TNF-α system in late antral follicles and, (4) the influence of TNF-α on cumulus cells expansion, ultrastructure and on expression of HAS2, CASP3 and CASP6 in follicular cells cultured for 24 h. An increased expression of TNF-α and TNFR1 was observed after 3, 6 and 12 h of GnRH treatment when compared to 0 and 24h. Higher TNFR2 mRNA levels were observed 3, 6 and 12 h after GnRH, when compared to 0 and 24 h. Proteins of TNF-α system were also expressed in late antral follicles. In vitro, TNF-α did not affect cumulus cells expansion, but reduced the HAS2, CASP3 and CASP6 mRNA levels in cumulus cells after 12 h. After 24 h of culture, TNF-α increased the mRNA levels for CASP6 in mural granulosa cells, while the TNF-α, TNFR1 and TNFR2 mRNA levels were increased in cumulus-oocyte complexes (COCs) cultured for 12 h with gonadotropins, but not after 24 h. Ultrastructural analysis confirmed the integrity of COCs cultured in presence of TNF-α. In conclusion, TNF-α system members are present in bovine antral follicles and expression of TNF-α is influenced by gonadotropins in vivo and in vitro. In vitro, TNF-α maintained cumulus cells ultrastructure during COC culture.

The effects of dehydroepiandrosterone on sexual function: a systematic review.

OBJECTIVE: Faced with the growing interest about the action of dehydroepiandrosterone (DHEA) and its benefits, as well as the negative impacts that sexual dysfunctions have on people's quality of life, this systematic review was undertaken with the objective of evaluating the effect of DHEA use on aspects of sexual function. METHOD: An electronic search was conducted in the databases of PubMed, ISI Web of Science and Virtual Health Library (VHL) combining the terms 'DHEA treatment' and 'DHEA use' with terms such as 'sexual dysfunction', 'sexual frequency' and 'libido'. No limits on time and language were imposed. Clinical studies were considered eligible where individuals for any reason made use of DHEA and if they had any aspect of sexual function assessed. Preclinical studies and systematic reviews were considered ineligible. RESULTS: The search identified 183 references and 38 were considered eligible. DHEA improved aspects such as sexual interest, lubrication, pain, arousal, orgasm and sexual frequency. Its effect was better in populations with sexual dysfunction, especially in perimenopausal and postmenopausal women. CONCLUSION: Considering the studies currently published, DHEA is effective in improving several aspects of sexual function, but this effect did not reach all the populations studied.

Isolates of Bacillus thuringiensis from Maranhão biomes with potential insecticidal action against Aedes aegypti larvae (Diptera, Culicidae)

Abstract Entomopathogenic agents are viable and effective options due to their selective action against insects but benign effects on humans and the environment. The most promising entomopathogens include subspecies of Bacillus thuringiensis (Bt), which are widely used for the biological control of insects, including mosquito vectors of human pathogens. The efficacy of B. thuringiensis toxicity has led to the search for new potentially toxic isolates in different regions of the world. Therefore, soil samples from the Amazon, Cerrado and Caatinga biomes of the state of Maranhão were evaluated for their potential larvicidal action against Aedes aegypti. The isolates with high toxicity to mosquito larvae, as detected by bioassays, were subjected to histological evaluation under a light microscope to identify the genes potentially responsible for the toxicity. Additionally, the toxic effects of these isolates on the intestinal epithelium were assessed. In the new B. thuringiensis isolates toxic to A. aegypti larvae, cry and cyt genes were amplified at different frequencies, with cry4, cyt1, cry32, cry10 and cry11 being the most frequent (33-55%) among those investigated. These genes encode specific proteins toxic to dipterans and may explain the severe morphological changes in the intestine of A. aegypti larvae caused by the toxins of the isolates.

Resumo Os agentes entomopatógenos são alternativas viáveis e eficazes, devido à sua ação seletiva para insetos sendo inofensivos ao homem e ao meio ambiente. Dentre os entomopatógenos mais promissores, destacam-se as subespécies de Bacillus thuringiensis (Bt) amplamente utilizadas no controle biológico de insetos incluindo espécies de mosquitos vetores de agentes patogênicos ao homem. A eficiência da toxicidade de Bt incentiva a prospecção de novos isolados em diversas regiões do mundo. Desta forma, em busca de novos isolados de B. thuringiensis potencialmente tóxicos, amostras de solo provenientes dos biomas Amazônia, Cerrado e Caatinga do estado do Maranhão foram avaliadas em relação ao seu potencial larvicida para Aedes aegypti. Os isolados que provocaram elevada toxicidade para larvas do mosquito, detectada por bioensaios, foram avaliados em relação aos potenciais genes responsáveis pela atividade tóxica, além da avaliação de efeitos tóxicos no epitélio intestinal através de análises histológicas em microscopia de luz. Os novos isolados de Bt tóxicos para larva de A. aegypti amplificaram frequências diferentes de genes cry e cyt sendo os mais frequentes (55-33%) os cry4, cyt1, cry32, cry10 e cry11 dentre os investigados. Esses genes codificam para proteínas tóxicas específicas para ordem Diptera, e podem explicar as severas alterações morfológicas provocadas pelas toxinas dos isolados observadas no intestino das larvas de A. aegypti.

The implant surface characteristics and peri-implantitis. An evidence-based update.

Peri-implantitis is an inflammatory disease of the peri-implant mucosa with the loss of supporting bone. Because of the absence of an un-inflamed connective tissue zone between the healthy and diseased sites, peri-implant lesions are thought to progress more rapidly than periodontal lesions, suggesting the importance of early diagnosis and intervention if possible. A number of risk factors have been identified that may lead to the initiation and progression of peri-implant mucositis and peri-implantitis, eg., previous periodontal disease, poor plaque control, inability to clean, residual cement, smoking, genetic factors, diabetes, occlusal overload, rheumatoid arthritis, increased time of loading and alcohol consumption. At present there is not much literature available, highlighting the relationship between implant surface characteristics and peri-implant diseases. Implant surface characteristics vary with respect to topography, roughness and clinical composition, including turned, blasted, acid etched, porous sintered, oxidized, plasma sprayed and hydroxyapatite coated surfaces and their combinations. So the aim of this review is to explore the relationship between the characteristics of implant surface, the prevalence and incidence of peri-implantitis. This would help to identify plausible influence of surface characteristics, oral hygiene instructions and maintenance of implants for the long-term uneventful success of implant therapy.

Expression of angiotensin II receptors in the caprine ovary and improvement of follicular viability in vitro.

This study aimed to evaluate mRNA levels of angiotensin II (ANG II) receptors (AGTR1 and AGTR2) in caprine follicles and to investigate the influence of ANG II on the viability and in vitro growth of preantral follicles. Real-time polymerase chain reaction (PCR) was used to quantify AGTR1 and AGTR2 mRNA levels in the different follicular stages. For culture, caprine ovaries were collected, cut into 13 fragments and then either directly fixed for histological and ultrastructural analysis (fresh control) or placed in culture for 1 or 7 days in α-minumum essential medium plus (α-MEM+) with 0, 1, 5, 10, 50 or 100 ng/ml ANG II. Then, the fragments were destined to morphological, viability and ultrastructural analysis. The results showed that primordial follicles had higher levels of AGTR1 and AGTR2 mRNA than secondary follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA than their respective cumulus-oocyte complex (COCs). After 7 days of culture, ANG II (10 or 50 ng/ml) maintained the percentages of normal follicles compared with α-MEM+. Fluorescence and ultrastructural microscopy confirmed follicular integrity in ANG II (10 ng/ml). In conclusion, a high expression of AGTR1 and AGTR2 is observed in primordial follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA. Finally, 10 ng/ml ANG II maintained the viability of caprine preantral follicles after in vitro culture.