Behavioral Despair Is Blocked by the Flavonoid Chrysin (5,7-Dihydroxyflavone) in a Rat Model of Surgical Menopause.

Women have a high susceptibility to the negative effects of stress. Hormonal changes experienced throughout their reproductive life partially contribute to a higher incidence of anxiety and depression symptoms, particularly, during natural or surgical menopause. In preclinical research, the flavonoid chrysin (5,7-dihydroxyflavone) exerts anxiolytic- and anti-despair-like effects; however, it is unknown whether chrysin exerts a protective effect against the behavioral changes produced by acute stress on locomotor activity and behavioral despair in rats at 12-weeks post-ovariectomy. Ovariectomized female Wistar rats were assigned to eight groups: vehicle group (10% DMSO), three groups with chrysin and three groups with the same dose of allopregnanolone (0.5, 1, and 2 mg/kg), and one group with diazepam (2 mg/kg). The treatments were administered for seven consecutive days and the effects were evaluated in the locomotor activity and swimming tests. Chrysin (2 mg/kg) increased the latency to first immobility and decreased the total immobility time in the swimming test as the reference drugs allopregnanolone and diazepam (2 mg/kg); while locomotor activity prevented the behavioral changes produced by swimming. In conclusion, chrysin exerts a protective effect against the behavioral changes induced by acute stress, similarly to the neurosteroid allopregnanolone and the benzodiazepine diazepam in rats subjected to a surgical menopause model.

Reproductive performance and luteal function of Santa Ines ewes inseminated by cervical retraction with fresh or frozen semen / Desempenho reprodutivo e função luteal de ovelhas Santa Inês inseminadas por retração cervical com sêmen fresco ou congelado

The artificial insemination (AI) by cervical retraction using fresh or commercially available frozen semen was evaluated. Santa Inês ewes (n=151) were assigned in five groups. Natural mating (NM) composed the control group. Four groups were submitted to hormonal treatment and timed insemination by cervical retraction (CRI) or laparoscopy (LAI), using fresh or frozen semen. To perform CRI, the ewes were kept in standing position. The time required to penetrate the cervical canal and to perform CRI were recorded; local deposition of semen and the reaction intensity of the ewe (weak, moderate or strong) were measured. Blood samples were collected to perform the progesterone dosage at days 0, 3, 5, 12 and 17 (day 0 = AI/NM). Non-return to estrus rate (NRE), pregnancy (at D35) and fertility (birth / mated ewes) were evaluated. Mean time for cervical penetration and CRI were 52.8 ± 21.2s and 3:26min ± 47s, respectively. Reaction intensity to CRI was weak or moderate in 92.3% of the ewes. Serum levels of progesterone after CRI, LAI or NM were similar. NRE of the inseminated ewes were similar, except for CRI with frozen semen, which was lower (P<0.05). Fertility rates were similar between CRI and LAI (35.4% and 42.2%, respectively). Artificial insemination by cervical retraction with ewes in a standing position is practical, and does not change the progesterone profile, providing reproductive rates similar to laparoscopy. However, the use of this technique along with commercial frozen semen presents low fertility rate.(AU)

Avaliou-se a inseminação artificial por retração cervical usando sêmen fresco ou congelado disponível comercialmente. Ovelhas Santa Inês foram divididas em cinco grupos. A monta natural (MN) correspondeu ao grupo controle. Quatro grupos foram submetidos a um tratamento hormonal e inseminação artificial por retração cervical (CRI) ou laparoscopia (LAI) usando sêmen fresco ou congelado. Para a CRI, ovelhas foram mantidas em estação. O tempo para penetrar o canal cervical e realizar a CRI foi mensurado, bem como o local de deposição do sêmen e intensidade de reação da ovelha (fraca, moderada, forte). Amostras de sangue foram coletadas para dosagem de progesterona aos dias 0, 3, 5, 12 e 17 (dia 0 = inseminação ou monta). Foram avaliadas as taxas de não retorno ao estro (NRE), gestação (ao dia 35) e fertilidade (nascimento / ovelhas acasaladas). O tempo médio para penetração cervical foi de 52,8 ± 21,2s e para a CRI foi de 3:26min ± 47s. A intensidade de reação à CRI foi fraca ou moderada em 92,3% das ovelhas. Os níveis séricos de progesterona após CRI, LAI e MN foram similares. NRE das ovelhas inseminadas foi similar, exceto em ovelhas com CRI e sêmen congelado, que foi inferior (P<0,05). As taxas de fertilidade foram semelhantes entre CRI e LAI (35,4% e 42,2%, respectivamente). A inseminação artificial por retração cervical com ovelhas em estação se mostrou prática e não alterou o perfil de progesterona, possibilitando índices reprodutivos similares à laparoscopia. No entanto, o uso desta técnica com sêmen disponível comercialmente apresentou baixa taxa de fertilidade.(AU)

Reproductive performance and luteal function of Santa Ines ewes inseminated by cervical retraction with fresh or frozen semen / Desempenho reprodutivo e função luteal de ovelhas Santa Inês inseminadas por retração cervical com sêmen fresco ou congelado

The artificial insemination (AI) by cervical retraction using fresh or commercially available frozen semen was evaluated. Santa Inês ewes (n=151) were assigned in five groups. Natural mating (NM) composed the control group. Four groups were submitted to hormonal treatment and timed insemination by cervical retraction (CRI) or laparoscopy (LAI), using fresh or frozen semen. To perform CRI, the ewes were kept in standing position. The time required to penetrate the cervical canal and to perform CRI were recorded; local deposition of semen and the reaction intensity of the ewe (weak, moderate or strong) were measured. Blood samples were collected to perform the progesterone dosage at days 0, 3, 5, 12 and 17 (day 0 = AI/NM). Non-return to estrus rate (NRE), pregnancy (at D35) and fertility (birth / mated ewes) were evaluated. Mean time for cervical penetration and CRI were 52.8 ± 21.2s and 3:26min ± 47s, respectively. Reaction intensity to CRI was weak or moderate in 92.3% of the ewes. Serum levels of progesterone after CRI, LAI or NM were similar. NRE of the inseminated ewes were similar, except for CRI with frozen semen, which was lower (P<0.05). Fertility rates were similar between CRI and LAI (35.4% and 42.2%, respectively). Artificial insemination by cervical retraction with ewes in a standing position is practical, and does not change the progesterone profile, providing reproductive rates similar to laparoscopy. However, the use of this technique along with commercial frozen semen presents low fertility rate.

Avaliou-se a inseminação artificial por retração cervical usando sêmen fresco ou congelado disponível comercialmente. Ovelhas Santa Inês foram divididas em cinco grupos. A monta natural (MN) correspondeu ao grupo controle. Quatro grupos foram submetidos a um tratamento hormonal e inseminação artificial por retração cervical (CRI) ou laparoscopia (LAI) usando sêmen fresco ou congelado. Para a CRI, ovelhas foram mantidas em estação. O tempo para penetrar o canal cervical e realizar a CRI foi mensurado, bem como o local de deposição do sêmen e intensidade de reação da ovelha (fraca, moderada, forte). Amostras de sangue foram coletadas para dosagem de progesterona aos dias 0, 3, 5, 12 e 17 (dia 0 = inseminação ou monta). Foram avaliadas as taxas de não retorno ao estro (NRE), gestação (ao dia 35) e fertilidade (nascimento / ovelhas acasaladas). O tempo médio para penetração cervical foi de 52,8 ± 21,2s e para a CRI foi de 3:26min ± 47s. A intensidade de reação à CRI foi fraca ou moderada em 92,3% das ovelhas. Os níveis séricos de progesterona após CRI, LAI e MN foram similares. NRE das ovelhas inseminadas foi similar, exceto em ovelhas com CRI e sêmen congelado, que foi inferior (P<0,05). As taxas de fertilidade foram semelhantes entre CRI e LAI (35,4% e 42,2%, respectivamente). A inseminação artificial por retração cervical com ovelhas em estação se mostrou prática e não alterou o perfil de progesterona, possibilitando índices reprodutivos similares à laparoscopia. No entanto, o uso desta técnica com sêmen disponível comercialmente apresentou baixa taxa de fertilidade.

The Step of Incorporation of Bacillus coagulans GBI-30 6086 Into "requeijão cremoso" Processed Cheese Does Not Affect Metabolic Homeostasis of Rats.

Dairy product consumption is a common habit in Brazil. These products present a good matrix for probiotic incorporation. Thus, in this study the feasibility of producing a probiotic "requeijão cremoso" incorporated with Bacillus coagulans GBI-30 6086 in three different steps and its metabolic effect in an animal model for 2 weeks has been evaluated. Wistar adult health rats were randomized into one to five groups (n = 8 for each group): Control (C); "requeijão cremoso" without probiotic (RC); probiotic inoculated in the milk before pasteurization at 65°C/30 min (RPP); "requeijão cremoso" inoculated before the fusion step and consequently exposed to 90°C/5 min (RPF); and "requeijão cremoso" inoculated after fusion step, i.e., once the product temperature reached 50°C (RPAF). At the end of treatment, analysis of molecular markers of proteins of stress and antioxidant system, HSP 25, 60, 70 and 90, SOD and catalase were performed in the animals' muscles by Western Blot technique. The HSP25, HSP90 and catalase levels of C, RPP, RPF, and RPAF were similar, indicating that the homeostasis remained unchanged. The incorporation of B. coagulans GBI-30 6086 in the "requeijão cremoso" was shown to be stable and the microorganism remained viable in all steps tested. The incorporation of the probiotic strain in the fusion stage facilitated the technological process, since it allowed a better homogenization of the product and did not affect the maintenance of the metabolic homeostasis of rats.

Effects of light intensity on growth, anti-stress ability and immune function in yellow feathered broilers

An experiment was conducted to investigate the effects of light intensity on growth, anti-stress ability, and immune function of yellow feathered broilers. A total of 480 one-day-old male Lingnan yellow feathered broilers were randomly allocated to 4 treatments based on light intensity (1, 5, 20 and 80 lx) with 8 replicates of 15 chicks each. The experiment lasted for 63 days. Compared with those under high light intensity, broilers exposed to low light intensity had higher (p<0.05) total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), a-Naphthylacetate esterase (ANAE+), antibody titer, but lower (p<0.05) malonaldehyde (MDA) levels and heterophil/lymphocyte ratio (H/L). There was a linear effect for T-AOC(p=0.002), GSH-Px(p≤0.047), MDA (p=0.003), H/L(p≤0.014), ANAE+ (p≤0.044), and antibody titer (p≤0.021) with T-AOC, GSH-Px, ANAE+, and antibody titer increased significantly as light intensity decreased, whereas MDA and H/L were decreased with the decrease in light intensity. These results suggested that broilers under low light intensity could have similar performance, better anti-stress ability, stronger immune function, and more efficient in energy usage as compared with those exposed to high light intensity environment.(AU)

Effects of light intensity on growth, anti-stress ability and immune function in yellow feathered broilers

An experiment was conducted to investigate the effects of light intensity on growth, anti-stress ability, and immune function of yellow feathered broilers. A total of 480 one-day-old male Lingnan yellow feathered broilers were randomly allocated to 4 treatments based on light intensity (1, 5, 20 and 80 lx) with 8 replicates of 15 chicks each. The experiment lasted for 63 days. Compared with those under high light intensity, broilers exposed to low light intensity had higher (p<0.05) total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), a-Naphthylacetate esterase (ANAE+), antibody titer, but lower (p<0.05) malonaldehyde (MDA) levels and heterophil/lymphocyte ratio (H/L). There was a linear effect for T-AOC(p=0.002), GSH-Px(p≤0.047), MDA (p=0.003), H/L(p≤0.014), ANAE+ (p≤0.044), and antibody titer (p≤0.021) with T-AOC, GSH-Px, ANAE+, and antibody titer increased significantly as light intensity decreased, whereas MDA and H/L were decreased with the decrease in light intensity. These results suggested that broilers under low light intensity could have similar performance, better anti-stress ability, stronger immune function, and more efficient in energy usage as compared with those exposed to high light intensity environment.

Royal jelly decreases corticosterone levels and improves the brain antioxidant system in restraint and cold stressed rats.

Restraint and cold stress induces the hypothalamic-pituitary-adrenal (HPA) axis to release corticosterone from the adrenal gland, which can worsen the antioxidant defense system in the central nervous system. Here, we investigated the corticosterone levels and the antioxidant defense system in the cerebellum and brain, as well as in its isolated regions, such as cerebral cortex, striatum and hippocampus of stressed rats supplemented with royal jelly (RJ). Wistar rats were supplemented with RJ for 14days and the stress induction started on the 7th day. Stressed rats increased corticosterone levels, glycemia and lipid peroxidation in the brain and cerebellum, cerebral cortex and hippocampus besides reduced glutathione defense system in the brain and striatum. Rats supplemented with RJ decreased corticosterone, maintained glycemia and decreased lipid peroxidation in the brain, cerebellum, as well as striatum and hippocampus, besides improved glutathione defense system in cerebral cortex and striatum. This study suggests an anti-stress and neuroprotective effect of RJ under stress conditions.

Timing of progesterone and allopregnanolone effects in a serial forced swim test / Duración del efecto de progesterona y alopregnanolona en la prueba de nado forzado seriada

The forced swim test (FST) is commonly employed to test the potency of drugs to reduce immobility as an indicator of anti-despair. Certainly, antidepressant drugs reduce the total time of immobility and enlarge the latency to the first immobility period. FST is preceded by the open field test (OFT) to discard any influence of changes in general motor activity that could interfere with immobility in the FST. Albeit progesterone and its a-reduced metabolite allopregnanolone produce antidepressant-like effects in the FST, the timing of actions is unknown. We hypothesized that the latency and duration of effects produced by progesterone and allopregnanolone may be characterized by repeated FST sessions; we therefore devised a serial-FST experimental design to evaluate the timing effects of these steroids on immobility, locomotion in the open field test, and grooming in the later as an indicator of response to stress. We included fifty-one ovariectomized adult Wistar rats weighing 200-250 g at the beginning of the experiments. They were ovariectomized by abdominal approach under anesthesia. Rats were housed six per cage, at room temperature (25 ± 1°C) under a 12 h/12 h light/dark cycle (lights ON at 7:00 a.m.) with ad libitum access to purified water and food. All of the experimental procedures followed National Institutes of Health Guidelines. The local Ethics Committee (Biomedical Research Institute, Universidad Nacional Autónoma de México) approved the experimental protocol. A first group received vehicle (2-hidroxypropyl-γ-cyclodextrin dissolved in injectable sterilized water to obtain a 35% solution, control group n=17), the second group progesterone (1.0 mg/kg, n=17), and the third group allopregnanolone (1.0 mg/kg, n=17). All single injections were applied by intraperitoneal route at a volume of 0.8 ml/kg. The effects of treatments were evaluated in the serial-FST at 0.25, 0.5, 1, 2, 4, 6, and 24 h after injection, in a rectangular pool (height, 60 cm; length, 30 cm; width, 50 cm), with 24 cm deep water (25 ± 1°C). We evaluated the total time of immobility, during 5 min, considered as the principal indicator of an anti-despair effect. Before each session of serial-FST, locomotion was evaluated in the OFT during 5 minutes. The apparatus consisted on an acrylic box (height, 20 cm; length, 44 cm; width, 33 cm), with twelve squares delineated on the floor (11x11 cm). In the same OFT sessions, grooming was evaluated as an indicator of response to stress. Statistical analysis consisted in two-way analysis of variance (ANOVA) and Student-Newman-Keuls as post hoc test. Total time in immobility was the highest and remained at similar levels only in the control group throughout the seven sessions of the serial-FST. In the allopregnanolone group a reduction in immobility was observed beginning 0.5 h after injection and lasted approximately 1.5 h. Similarly, progesterone reduced immobility beginning 1.0 h after injection, and the reduction lasted for approximately 5.0 h. In all groups, locomotion in the OFT was reduced after the first serial-FST session and remained at similar low levels during the serial-FST. In the control group, grooming was reduced after the first serial-FST session and lasted 24 h, but grooming did not change in the progesterone-or allopregnanolone-treated rats. From a serial-FST design, we conclude that progesterone and allopregnanolone exert short time-dependent reductions in immobility and anti-stress-like effects no longer than 24 hrs, and seemingly a reduction in the response to stress, which may have some clinical applications.

Introducción La progesterona y su metabolito activo alopregnanolona se han estudiado ampliamente en modelos experimentales de ansiedad y depresión, y por su propiedad de ser sintetizadas en el cerebro se les considera como neuroesteroides. Entre las pruebas que permiten determinar la potencia antidepresiva de ciertos fármacos se encuentra la prueba de nado forzado, la cual se diseñó originalmente para detectar la potencia de sustancias con propiedades antidepresivas. Estas sustancias reducen el tiempo de inmovilidad y alargan la latencia al primer periodo de inmovilidad, lo cual es considerado como un efecto antidepresivo. Usualmente, la prueba de nado forzado se aplica dos veces, una sesión de preprueba que dura 15 minutos, en la cual la rata o ratón desarrolla el estado de desesperanza. La preprueba es seguida de la sesión de prueba que se realiza 24 horas después durante 5 minutos. En ella se evalúa el efecto de las sustancias con propiedades antidepresivas. Además, la prueba de nado forzado es precedida por la prueba de campo abierto con la finalidad de identificar cambios en la actividad motora general (hipoactividad o hiperactividad) que pudiera interferir con la interpretación de las variables evaluadas en la prueba de nado forzado. Algunos esteroides, como la progesterona y alopregnanolona, reducen la inmovilidad y alargan la latencia a la primera inmovilidad en la prueba de nado forzado, lo que indica su efecto tipo-antidepresivo. Sin embargo, la latencia y la duración de los efectos farmacológicos son desconocidas. La hipótesis de este trabajo fue que, si utilizábamos la prueba de nado forzado de forma repetida, podríamos identificar el tiempo de duración de los efectos de estos esteroides. Por lo tanto, diseñamos un experimento con la prueba de nado forzado seriada para evaluar el tiempo de permanencia de los efectos de progesterona y alopregnanolona en esta prueba conductual. Materiales y métodos Sujetos: En este estudio se incluyeron 51 ratas adultas ovariectomizadas de la cepa Wistar, con peso entre 200 y 250 g al inicio de los experimentos. Las ratas fueron anestesiadas y ovariectomizadas por aproximación ventral y fueron alojadas en cajas de acrílico trasparente (n=6), con una temperatura ambiente de 25 ± 1°C y con un ciclo de luz-oscuridad de 12 ×12 h (la luz se encendió a las 7:00 am). Las ratas tuvieron libre acceso al agua purificada y al alimento (Purina). Todos los procedimientos realizados en este estudio fueron de acuerdo con las normas éticas en el uso de animales de experimentación, basándonos en la Guía del National Institute of Health, y el protocolo fue aprobado por el Comité de Ética del Instituto de Investigaciones Biomédicas de la Universidad Nacional Autónoma de México. Grupos y tratamientos: Las ratas del grupo control recibieron el vehículo (solución al 35% de 2-hidroxipropil-g-ciclodextrina), el segundo grupo recibió progesterona (1.0 mg/kg) y el tercero recibió alopregnanolona (1.0 mg/kg) por vía intraperitoneal, en un volumen de 0.8 ml/kg. Pruebas conductuales: El efecto de los tratamientos fue evaluado en la prueba de nado forzado a las 0.25, 0.5, 1, 2, 4, 6 y 24 horas después de la administración. Utilizamos un estanque rectangular (base 50 × 34 cm, altura 60 cm), con agua a 25°C y una altura de 24 cm. Sólo se evaluó el tiempo total de inmovilidad, considerando que es el principal indicador de un efecto antidesesperanza. Antes de cada sesión de nado forzado se evaluó la actividad motora (cuadros deambulados) y el acicalamiento en campo abierto. Esta prueba consistió en colocar a la rata en una caja de acrílico (base 33 × 44 cm, altura 20 cm) con el piso dividido en 12 cuadros de 11 × 11 cm. Los resultados obtenidos de ambas pruebas fueron evaluados por medio de una ANOVA de dos vías y como prueba post hoc se aplicó Student-Newman-Keuls. Resultados La prueba de nado forzado aplicada de forma repetida resultó ser útil para evaluar los efectos temporales producidos por dos esteroides con potencia antidepresiva. Las ratas del grupo control mostraron los valores más altos de inmovilidad en la prueba de nado forzado, los cuales se mantuvieron así durante las sesiones de prueba. En los grupos tratados con progesterona o alopregnanolona hubo una reducción de la inmovilidad, gradual y temporal. Los animales tratados con alopregnanolona redujeron la inmovilidad a partir de las 0.5 horas después de la administración, efecto que se mantuvo por un periodo de 1.5 h. Los animales tratados con progesterona redujeron la inmovilidad a partir de 1.0 hora después de la administración, efecto que se mantuvo por un periodo de 5.0h. En campo abierto, independientemente del tratamiento, hubo una reducción del número de cuadros cruzados después de la primera sesión de nado forzado, efecto que permaneció hasta las 24h. En el acicalamiento, se observó que sólo los animales del grupo control redujeron significativamente el tiempo empleado en esta conducta, mientras que los animales inyectados con progesterona o alopregnanolona no modificaron esta variable. Es decir, mantuvieron niveles semejantes durante todas las sesiones de prueba y estuvieron por arriba de los valores encontrados en los animales control. Conclusión La progesterona y la alopregnanolona ejercen un efecto antidesesperanza de breve latencia, no mayor a 24 horas. Este hallazgo podría tener implicaciones clínicas en pacientes con depresión refractaria al tratamiento convencional.

Tonic, fortifier and aphrodisiac: adaptogens in the Brazilian folk medicine

In Brazil, many plants are used as tonic, fortifier, aphrodisiac, anti-stress, among other uses that are similar to the indications of an adaptogen. In general, such plants are used unspecifically, in situations of stress and fatigue, in the recovery after a previous pathological or debilitating state, or simply aiming at the maintenance of a healthy state. This article discusses the popular terms employed in the Brazilian folk medicine for the plants with this profile, their particularities and limitations. The article also discusses the possible mechanisms of action of an adaptogen and compares the main Brazilian plants used for that purpose: guarana (Paullinia cupana Kunth, family Sapindaceae), muirapuama (Ptychopetalum olacoides Benth., Olacaceae), catuaba (Anemopaegma arvense (Vell.) Stellfeld & J.F. Souza, Bignoniaceae, and Trichilia catigua A. Juss., Meliaceae), nó-decachorro (Heteropterys aphrodisiaca O. Mach, Malpighiaceae), damiana (Turnera diffusa Willd. ex Schult., Turneraceae) and pfaffia or Brazilian ginseng (Pfaffia sp, Amaranthaceae).