Curcumin exerts its antioxidant and neuroprotective effects against aluminum-induced oxidative stress and neurotoxicity in male albino rats

Aluminum is a neurotoxicant and one of the most harmful metals in the environment; it is producing tissue inflammation and oxidative stress. Curcumin is an effective antioxidant and neuroprotective compound with medicinal potential. Curcumin's effect on AL toxicity was investigated in this study. Two groups of 70 mature adult albino rats were used, each of which was subdivided into five groups: Control, Vehicle, Curcumin, Aluminum, and Curcumin + Aluminum group. For two periods of 20 and 40 days, animal models were administered orally AlCl3 (20 mg kg-1 bw) and/or Curcumin (100 mg kg-1 bw). In the cerebral cortex, aluminum caused a significant rise (p < 0.05) in lipid peroxidation and DNA fragmentation, as well as a significant decrease (p < 0.05) in antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase. In the brain hippocampus, aluminum caused a major reduction (p < 0.05) in neurotransmitters (dopamine and serotonin), while Acetylcholine esterase activity increased sharply (p < 0.05). Aluminum also triggered histological analyses in the hippocampus of the brain. Curcumin co-administration considerably reduced the increase in lipid peroxidation and DNA fragmentation, but also enhanced the depletion of antioxidant enzymes. Curcumin also reversed the decline in neurotransmitters, the increase in Acetylcholine esterase, and the distortion in the brain hippocampus.(AU)

Non-Status Epilepticus female rats show seizure-like behaviors in the chronic phase of Pilocarpine experimental model

Abstract Only few studies have focus on animals that received Pilocarpine (Pilo) and did not develop behavioral status epilepticus (SE) and, whether they may become epileptic in the models chronic phase. Previews works observed mossy fiber sprouting in the hippocampus of Non-SE (NSE) rats, while others observed spontaneous and recurrent seizures (SRS) 6 - 8 months after animals received Pilo. It is known that neuronal excitability is influenced by female hormones, as well as, the occurrence of SE in castrated and non-castrated female rats. However, it is not known whether females that received Pilo and did not show SE, may have SRS. The aim of this work was to investigate whether castrated and non-castrated female rats that did not show behavioral SE after Pilo, will develop SRS in the following one-year. For that, animals received 360 mg/kg of Pilo and were video monitored for 12 months. SE females from castrated and non-castrated groups became epileptic since the first month after drug injection. Epileptic behaviors were identified watching video monitoring recordings in the fast speed. Castrated and Non-castrated NSE animals showed behaviors resembling seizures described by Racine Scale stages 1 - 3. Motor alterations showed by NSE groups could be observed only when recordings were analyzed in slow speed. In addition, behavioral manifestations as, rhythmic head movements, sudden head movements, whole body movements and immobility were also observed in both, SE and NSE groups. We concluded that NSE female rats may have become epileptic. Adding to it, slow speed analysis of motor alterations was essential for the observation of NSE findings, which suggests that possibly many motor alterations have been underestimated in epilepsy experimental research.

Resumo Poucos são os estudos com foco em animais que receberam Pilocarpina (Pilo) e não desenvolveram status epilepticus (SE) comportamental e, se os mesmos se tornarão epilépticos na fase crônica do modelo. Autores observaram o brotamento das fibras musgosas no hipocampo de ratos Não-SE (NSE), enquanto outros observaram crises espontâneas e recorrentes (CER) 6 - 8 meses após receberam a droga. A excitabilidade neuronal é influenciada pelos hormônios femininos e, da mesma forma, a ocorrência de SE em ratas castradas e não-castradas. Entretanto, não é sabido se as fêmeas que não apresentam SE terão CER. O objetivo deste trabalho foi investigar se fêmeas castradas e não castradas que não tiveram SE comportamental após a injeção de Pilo desenvolverão CER dentro de um ano. Para isto, os animais receberam 360 mg/kg de Pilo e foram videomonitorados por 12 meses. As fêmeas SE castradas e não-castradas se tornaram epilépticas desde o primeiro mês pós Pilo. O comportamento epiléptico foi identificado assistindo as gravações na velocidade rápida. As fêmeas NSE castradas e não-castradas apresentaram comportamentos similares aos estágios 1 - 3 da Escala de Racine. As alterações motoras nestes grupos (NSE) foram observadas apenas quando as videomonitoração foi analisada na velocidade lenta. Além destas, manifestações comportamentais como movimentos rítmicos da cabeça, movimentos súbitos da cabeça, movimentos de todo o corpo e imobilidade também foram observadas em ambos grupos, SE e NSE. Concluímos que as fêmeas NE podem ter se tornado epilépticas. Adicionado a isto, a análise das alterações motoras na velocidade lenta foi essencial para a observação dos achados das fêmeas NSE, o que sugere que possivelmente muitas alterações motoras têm sido subestimados na pesquisa em epilepsia experimental.

Non-Status Epilepticus female rats show seizure-like behaviors in the chronic phase of Pilocarpine experimental model / Ratas sem Status Epilepticus apresentam comportamento do tipo crise epiléptica na fase crônica do modelo experimental da Pilocarpina

Only few studies have focus on animals that received Pilocarpine (Pilo) and did not develop behavioral status epilepticus (SE) and, whether they may become epileptic in the model's chronic phase. Previews works observed mossy fiber sprouting in the hippocampus of Non-SE (NSE) rats, while others observed spontaneous and recurrent seizures (SRS) 6 - 8 months after animals received Pilo. It is known that neuronal excitability is influenced by female hormones, as well as, the occurrence of SE in castrated and non-castrated female rats. However, it is not known whether females that received Pilo and did not show SE, may have SRS. The aim of this work was to investigate whether castrated and non-castrated female rats that did not show behavioral SE after Pilo, will develop SRS in the following one-year. For that, animals received 360 mg/kg of Pilo and were video monitored for 12 months. SE females from castrated and non-castrated groups became epileptic since the first month after drug injection. Epileptic behaviors were identified watching video monitoring recordings in the fast speed. Castrated and Non castrated NSE animals showed behaviors resembling seizures described by Racine Scale stages 1 - 3. Motor alterations showed by NSE groups could be observed only when recordings were analyzed in slow speed. In addition, behavioral manifestations as, rhythmic head movements, sudden head movements, whole body movements and immobility were also observed in both, SE and NSE groups. We concluded that NSE female rats may have become epileptic. Adding to it, slow speed analysis of motor alterations was essential for the observation of NSE findings, which suggests that possibly many motor alterations have been underestimated in epilepsy experimental research.

Poucos são os estudos com foco em animais que receberam Pilocarpina (Pilo) e não desenvolveram status epilepticus (SE) comportamental e, se os mesmos se tornarão epilépticos na fase crônica do modelo. Autores observaram o brotamento das fibras musgosas no hipocampo de ratos Não-SE (NSE), enquanto outros observaram crises espontâneas e recorrentes (CER) 6 - 8 meses após receberam a droga. A excitabilidade neuronal é influenciada pelos hormônios femininos e, da mesma forma, a ocorrência de SE em ratas castradas e não-castradas. Entretanto, não é sabido se as fêmeas que não apresentam SE terão CER. O objetivo deste trabalho foi investigar se fêmeas castradas e não castradas que não tiveram SE comportamental após a injeção de Pilo desenvolverão CER dentro de um ano. Para isto, os animais receberam 360 mg/kg de Pilo e foram videomonitorados por 12 meses. As fêmeas SE castradas e não-castradas se tornaram epilépticas desde o primeiro mês pós Pilo. O comportamento epiléptico foi identificado assistindo as gravações na velocidade rápida. As fêmeas NSE castradas e não-castradas apresentaram comportamentos similares aos estágios 1 - 3 da Escala de Racine. As alterações motoras nestes grupos (NSE) foram observadas apenas quando as videomonitoração foi analisada na velocidade lenta. Além destas, manifestações comportamentais como movimentos rítmicos da cabeça, movimentos súbitos da cabeça, movimentos de todo o corpo e imobilidade também foram observadas em ambos grupos, SE e NSE. Concluímos que as fêmeas NE podem ter se tornado epilépticas. Adicionado a isto, a análise das alterações motoras na velocidade lenta foi essencial para a observação dos achados das fêmeas NSE, o que sugere que possivelmente muitas alterações motoras têm sido subestimados na pesquisa em epilepsia experimental.

Non-Status Epilepticus female rats show seizure-like behaviors in the chronic phase of Pilocarpine experimental model / Ratas sem Status Epilepticus apresentam comportamento do tipo crise epiléptica na fase crônica do modelo experimental da Pilocarpina

Only few studies have focus on animals that received Pilocarpine (Pilo) and did not develop behavioral status epilepticus (SE) and, whether they may become epileptic in the model's chronic phase. Previews works observed mossy fiber sprouting in the hippocampus of Non-SE (NSE) rats, while others observed spontaneous and recurrent seizures (SRS) 6 - 8 months after animals received Pilo. It is known that neuronal excitability is influenced by female hormones, as well as, the occurrence of SE in castrated and non-castrated female rats. However, it is not known whether females that received Pilo and did not show SE, may have SRS. The aim of this work was to investigate whether castrated and non-castrated female rats that did not show behavioral SE after Pilo, will develop SRS in the following one-year. For that, animals received 360 mg/kg of Pilo and were video monitored for 12 months. SE females from castrated and non-castrated groups became epileptic since the first month after drug injection. Epileptic behaviors were identified watching video monitoring recordings in the fast speed. Castrated and Non castrated NSE animals showed behaviors resembling seizures described by Racine Scale stages 1 - 3. Motor alterations showed by NSE groups could be observed only when recordings were analyzed in slow speed. In addition, behavioral manifestations as, rhythmic head movements, sudden head movements, whole body movements and immobility were also observed in both, SE and NSE groups. We concluded that NSE female rats may have become epileptic. Adding to it, slow speed analysis of motor alterations was essential for the observation of NSE findings, which suggests that possibly many motor alterations have been underestimated in epilepsy experimental research.(AU)

Poucos são os estudos com foco em animais que receberam Pilocarpina (Pilo) e não desenvolveram status epilepticus (SE) comportamental e, se os mesmos se tornarão epilépticos na fase crônica do modelo. Autores observaram o brotamento das fibras musgosas no hipocampo de ratos Não-SE (NSE), enquanto outros observaram crises espontâneas e recorrentes (CER) 6 - 8 meses após receberam a droga. A excitabilidade neuronal é influenciada pelos hormônios femininos e, da mesma forma, a ocorrência de SE em ratas castradas e não-castradas. Entretanto, não é sabido se as fêmeas que não apresentam SE terão CER. O objetivo deste trabalho foi investigar se fêmeas castradas e não castradas que não tiveram SE comportamental após a injeção de Pilo desenvolverão CER dentro de um ano. Para isto, os animais receberam 360 mg/kg de Pilo e foram videomonitorados por 12 meses. As fêmeas SE castradas e não-castradas se tornaram epilépticas desde o primeiro mês pós Pilo. O comportamento epiléptico foi identificado assistindo as gravações na velocidade rápida. As fêmeas NSE castradas e não-castradas apresentaram comportamentos similares aos estágios 1 - 3 da Escala de Racine. As alterações motoras nestes grupos (NSE) foram observadas apenas quando as videomonitoração foi analisada na velocidade lenta. Além destas, manifestações comportamentais como movimentos rítmicos da cabeça, movimentos súbitos da cabeça, movimentos de todo o corpo e imobilidade também foram observadas em ambos grupos, SE e NSE. Concluímos que as fêmeas NE podem ter se tornado epilépticas. Adicionado a isto, a análise das alterações motoras na velocidade lenta foi essencial para a observação dos achados das fêmeas NSE, o que sugere que possivelmente muitas alterações motoras têm sido subestimados na pesquisa em epilepsia experimental.(AU)

Effects of ortho-eugenol on anxiety, working memory and oxidative stress in mice / Efeitos do orto-eugenol na ansiedade, memória de trabalho e estresse oxidativo em camundongos

Ortho-eugenol is a synthetic derivative from eugenol, the major compound of clove essential oil, which has demonstrated antidepressant and antinociceptive effects in pioneering studies. Additionally, its effects appear to be dependent on the noradrenergic and dopaminergic systems. Depression and anxiety disorders are known to share a great overlap in their pathophysiology, and many drugs are effective in the treatment of both diseases. Furthermore, high levels of anxiety are related to working memory deficits and increased oxidative stress. Thus, in this study we investigated the effects of acute treatment of ortho-eugenol, at 50, 75 and 100 mg/kg, on anxiety, working memory and oxidative stress in male Swiss mice. Our results show that the 100 mg/kg dose increased the number of head-dips and reduced the latency in the hole-board test. The 50 mg/kg dose reduced malondialdehyde levels in the prefrontal cortex and the number of Y-maze entries compared to the MK-801-induced hyperlocomotion group. All doses reduced nitrite levels in the hippocampus. It was also possible to assess a statistical correlation between the reduction of oxidative stress and hyperlocomotion after the administration of ortho-eugenol. However, acute treatment was not able to prevent working memory deficits. Therefore, the present study shows that ortho-eugenol has an anxiolytic and antioxidant effect, and was able to prevent substance-induced hyperlocomotion. Our results contribute to the elucidation of the pharmacological profile of ortho-eugenol, as well as to direct further studies that seek to investigate its possible clinical applications.

Ortho-eugenol é um derivado sintético do eugenol, composto principal do óleo essencial de cravo, que demonstrou efeitos antidepressivos e antinociceptivos em estudos pioneiros. Além disso, seus efeitos parecem ser dependentes dos sistemas noradrenérgico e dopaminérgico. Sabe-se que os transtornos de depressão e ansiedade compartilham uma grande sobreposição em sua fisiopatologia, e muitas drogas são eficazes no tratamento de ambas as doenças. Além disso, altos níveis de ansiedade estão relacionados a déficits de memória de trabalho e aumento do estresse oxidativo. Assim, o presente estudo investigou os efeitos do tratamento agudo de orto-eugenol, nas doses de 50, 75 e 100 mg/kg, na ansiedade, memória de trabalho e estresse oxidativo em camundongos Swiss machos. Nossos resultados mostram que a dose de 100 mg/kg aumentou o número de mergulhos e reduziu a latência no teste da placa perfurada. A dose de 50 mg/kg reduziu os níveis de malondialdeído no córtex pré-frontal e o número de entradas no labirinto em Y em comparação com o grupo de hiperlocomoção induzida por MK-801. Todas as doses reduziram os níveis de nitrito no hipocampo. Também foi possível avaliar uma correlação estatística entre a redução do estresse oxidativo e a hiperlocomoção após a administração do orto-eugenol. No entanto, o tratamento agudo não foi capaz de prevenir os déficits de memória de trabalho. Portanto, o presente estudo mostra que o orto-eugenol tem efeito ansiolítico e antioxidante, sendo capaz de prevenir a hiperlocomoção induzida pela substância. Nossos resultados contribuem para a elucidação do perfil farmacológico do orto-eugenol, bem como para direcionar novos estudos que busquem investigar suas possíveis aplicações clínicas.

Non-Status Epilepticus female rats show seizure-like behaviors in the chronic phase of Pilocarpine experimental model / Ratas sem Status Epilepticus apresentam comportamento do tipo crise epiléptica na fase crônica do modelo experimental da Pilocarpina

Abstract Only few studies have focus on animals that received Pilocarpine (Pilo) and did not develop behavioral status epilepticus (SE) and, whether they may become epileptic in the model's chronic phase. Previews works observed mossy fiber sprouting in the hippocampus of Non-SE (NSE) rats, while others observed spontaneous and recurrent seizures (SRS) 6 - 8 months after animals received Pilo. It is known that neuronal excitability is influenced by female hormones, as well as, the occurrence of SE in castrated and non-castrated female rats. However, it is not known whether females that received Pilo and did not show SE, may have SRS. The aim of this work was to investigate whether castrated and non-castrated female rats that did not show behavioral SE after Pilo, will develop SRS in the following one-year. For that, animals received 360 mg/kg of Pilo and were video monitored for 12 months. SE females from castrated and non-castrated groups became epileptic since the first month after drug injection. Epileptic behaviors were identified watching video monitoring recordings in the fast speed. Castrated and Non-castrated NSE animals showed behaviors resembling seizures described by Racine Scale stages 1 - 3. Motor alterations showed by NSE groups could be observed only when recordings were analyzed in slow speed. In addition, behavioral manifestations as, rhythmic head movements, sudden head movements, whole body movements and immobility were also observed in both, SE and NSE groups. We concluded that NSE female rats may have become epileptic. Adding to it, slow speed analysis of motor alterations was essential for the observation of NSE findings, which suggests that possibly many motor alterations have been underestimated in epilepsy experimental research.

Resumo Poucos são os estudos com foco em animais que receberam Pilocarpina (Pilo) e não desenvolveram status epilepticus (SE) comportamental e, se os mesmos se tornarão epilépticos na fase crônica do modelo. Autores observaram o brotamento das fibras musgosas no hipocampo de ratos Não-SE (NSE), enquanto outros observaram crises espontâneas e recorrentes (CER) 6 - 8 meses após receberam a droga. A excitabilidade neuronal é influenciada pelos hormônios femininos e, da mesma forma, a ocorrência de SE em ratas castradas e não-castradas. Entretanto, não é sabido se as fêmeas que não apresentam SE terão CER. O objetivo deste trabalho foi investigar se fêmeas castradas e não castradas que não tiveram SE comportamental após a injeção de Pilo desenvolverão CER dentro de um ano. Para isto, os animais receberam 360 mg/kg de Pilo e foram videomonitorados por 12 meses. As fêmeas SE castradas e não-castradas se tornaram epilépticas desde o primeiro mês pós Pilo. O comportamento epiléptico foi identificado assistindo as gravações na velocidade rápida. As fêmeas NSE castradas e não-castradas apresentaram comportamentos similares aos estágios 1 - 3 da Escala de Racine. As alterações motoras nestes grupos (NSE) foram observadas apenas quando as videomonitoração foi analisada na velocidade lenta. Além destas, manifestações comportamentais como movimentos rítmicos da cabeça, movimentos súbitos da cabeça, movimentos de todo o corpo e imobilidade também foram observadas em ambos grupos, SE e NSE. Concluímos que as fêmeas NE podem ter se tornado epilépticas. Adicionado a isto, a análise das alterações motoras na velocidade lenta foi essencial para a observação dos achados das fêmeas NSE, o que sugere que possivelmente muitas alterações motoras têm sido subestimados na pesquisa em epilepsia experimental.

Behavioral and structural changes in the hippocampus of wistar epileptic rats are minimized by acupuncture associated or not with phenobarbital / Alterações comportamentais e estruturais do hipocampo de ratos Wistar epilépticos são minimizadas pela acupuntura em associação ou não com fenobarbital

The aim of this study was to analyze the behavior and histopathological changes in the hippocampus of epileptic Wistar rats treated with acupuncture associated or not with phenobarbital. The experiment used 44 male rats with 90 days of birth, induced to status epileptics with pilocarpine hydrochloride in a single dose of 350mg/kg, separated into treatment groups and submitted for 5 minutes to the elevated plus-maze test. Group 1 received 0.2mL of saline solution orally; Group 2 treated with acupuncture at the yintang, baihui, shishencong, jizhong, naohu, thianzu points; Group 3 received orally phenobarbital, daily dose of 20mg/kg; Group 4 treated with an association of acupuncture and oral phenobarbital; Group 5 random needling. The results obtained showed that Groups 2 (acupuncture) and 4 (acupuncture and phenobarbital) presented decreased anxiety, epileptic seizures, and neuronal death in the CA1, CA3 areas of the hippocampus when compared to animals in groups 1, 3 and 5. It is concluded that the association of phenobarbital and acupuncture points used in the experiment allowed for the control of epileptic seizures, reduction of anxiety and reduction of lesions in the subareas of the hippocampus.

O objetivo deste estudo foi analisar o comportamento e as alterações histopatológicas no hipocampo de ratos Wistar epilépticos tratados com acupuntura associada ou não a fenobarbital. O experimento utilizou 44 ratos machos, com 90 dias de nascimento, induzidos ao status epileticus com cloridrato de pilocarpina, em dose única de 350mg/kg, separados em grupos de tratamento e submetidos por cinco minutos ao teste de labirinto em cruz elevado. O grupo 1 recebeu, por via oral, 0,2mL de solução salina; o grupo 2 foi tratado com acupuntura nos pontos yintang, baihui, shishencong, jizhong, naohu, thianzu; o grupo 3 recebeu, por via oral, fenobarbital, dose diária de 20mg/kg; o grupo 4 foi tratado com associação acupuntura e fenobarbital por via oral; o grupo 5 recebeu agulhamento aleatório. Os resultados obtidos demonstraram que os grupos 2 (acupuntura) e 4 (acupuntura e fenobarbital) apresentaram diminuição da ansiedade, das crises epilépticas e da morte neuronal nas áreas CA1, CA3 do hipocampo quando comparados aos animais dos grupos 1, 3 e 5. Conclui-se que a associação do fenobarbital e dos pontos de acupuntura utilizados no experimento permitiu o controle das crises epilépticas, a redução da ansiedade e a diminuição das lesões nas subáreas do hipocampo.

The Unicorn Mouse: Cranial Lipoma in a B6. Cg Mouse

Intracranial lipomas, also called intracranial lipomatous hamartomas, have been reported in some strains of research mice, but are rare in C57BL6 strains. It is presumed based on current publications that this is the first report of an intracranial lipoma in a mouse with this genetic change (B6.Cg-Cnpy2 tm1.2 Zhli Alb-Cre). Grossly, a fur-covered, soft, cylindrical, exophytic mass on the dorsal midline of the cranium was evident. Upon dissection, a soft, white, tubular structure extended through a 1mm defect in the sagittal suture of the skull to the deep surface of the hypodermis. Histologically, the mass consisted of well demarcated proliferation of mature white adipocytes, each containing one large fat droplet. The mass extended from the cerebrum at the level just caudal to the hippocampus in the third ventricle, between the superior colliculus and the caudal portion of the retrosplenial area and through the sagittal suture to the hypodermis and was surrounded by normal brain tissue.(AU)

Application of active acoustic technology to assess the target strength of seahorses based on the presence of a reproductive organ

This study aims to provide reliable information on comparing the target strength (TS) values of seahorses based on the reproductive state. This study was carried out using hydro-acoustic technology in a water tank environment. Data were obtained using the single-beam scientific echo-sounder SIMRAD EK-15 at a frequency of 200 kHz and analyzed via the Sonar-4 software. The measurement result of the TS (mean±S.E.M.) of Hippocampus kuda female, male and pregnant male seahorses were -56.24±0.047, -57.25±0.032, and -58.26±0.06, respectively. There was a significant difference in the mean TS value of H. kuda based on the reproductive state (P < 0.05). Furthermore, the response to the mean TS value of pregnant H. kuda male (the presence of a brood pouch) was lowly significant compared to the female (absence of a brood pouch) (P< 0.05). The results showed a possibility of finding a pregnant H. kuda male in a water column through active acoustic methods in the future.(AU)

Application of active acoustic technology to assess the target strength of seahorses based on the presence of a reproductive organ

This study aims to provide reliable information on comparing the target strength (TS) values of seahorses based on the reproductive state. This study was carried out using hydro-acoustic technology in a water tank environment. Data were obtained using the single-beam scientific echo-sounder SIMRAD EK-15 at a frequency of 200 kHz and analyzed via the Sonar-4 software. The measurement result of the TS (mean±S.E.M.) of Hippocampus kuda female, male and pregnant male seahorses were -56.24±0.047, -57.25±0.032, and -58.26±0.06, respectively. There was a significant difference in the mean TS value of H. kuda based on the reproductive state (P < 0.05). Furthermore, the response to the mean TS value of pregnant H. kuda male (the presence of a brood pouch) was lowly significant compared to the female (absence of a brood pouch) (P< 0.05). The results showed a possibility of finding a pregnant H. kuda male in a water column through active acoustic methods in the future.(AU)

Application of active acoustic technology to assess the target strength of seahorses based on the presence of a reproductive organ

This study aims to provide reliable information on comparing the target strength (TS) values of seahorses based on the reproductive state. This study was carried out using hydro-acoustic technology in a water tank environment. Data were obtained using the single-beam scientific echo-sounder SIMRAD EK-15 at a frequency of 200 kHz and analyzed via the Sonar-4 software. The measurement result of the TS (mean±S.E.M.) of Hippocampus kuda female, male and pregnant male seahorses were -56.24±0.047, -57.25±0.032, and -58.26±0.06, respectively. There was a significant difference in the mean TS value of H. kuda based on the reproductive state (P < 0.05). Furthermore, the response to the mean TS value of pregnant H. kuda male (the presence of a brood pouch) was lowly significant compared to the female (absence of a brood pouch) (P< 0.05). The results showed a possibility of finding a pregnant H. kuda male in a water column through active acoustic methods in the future.

Anabolic steroids and their effects of on neuronal density in cortical areas and hippocampus of mice / Esteroides anabolizantes e seus efeitos na densidade neuronal em áreas corticais e no hipocampo de camundongos

Anabolic substances have been increasingly used by bodybuilders and athletes with the goal of improving performance and aesthetics. However, this practice has caused some concern to physicians and researchers because of unknowledge of consequences that the indiscriminate and illicit use of these substances can cause. Thus, this study analyzed the effects of two commercially available anabolic steroids (AS), Winstrol Depot® (Stanozolol) and Deposteron® (Testosterone Cypionate), in the neuronal density of limbic, motor and sensory regions on the cerebral cortex and in CA1, CA2, CA3 regions of the hippocampus. A total of 60 Swiss mice were used (30 males and 30 females), separated into three groups: control and two experimental groups, which received the AAS. From each brain, homotypic and semi-serial samples were taken in frontal sections from areas established for the study. The results showed that females treated with testosterone cypionate presented a reduction in all regions tested and the ones treated with Stanozolol showed a decrease in some hippocampal areas. Regarding male animals, stanozolol led to a decrease in neuron number in one hippocampal region. These data allow us to conclude that supra-physiological doses of steroids used in this study, can cause considerable damage to nervous tissue with ultrastructural and consequently behavioral impairment. These changes could interfere with the loss of physical yield and performance of athletes and non-athletes and may cause irreparable damage to individuals making irresponsible use of anabolic steroids.(AU)

As substancias anabólicas tem sido cada vez mais utilizadas por fisiculturistas e atletas com o objetivo de melhorar o desempenho e a estética. No entanto, essa prática tem causado algumas preocupações aos médicos e pesquisadores, devido ao desconhecimento das consequencias que o uso indiscriminado e ilícito dessas substâncias podem causar. Diante disso, este estudo analisou os efeitos de dois esteroides anabolizantes (EA) comercialmente disponíveis, Winstrol Depot® (Stanozolol) e Deposteron® (cipionato de testosterona), na densidade neuronal das regiões corticais límbica, motora e sensitive bem como das áreas CA1, CA2, CA3 do hipocampo. Foram utilizados 60 camundongos Swiss (30 machos e 30 fêmeas), separados em três grupos: controle e dois grupos experimentais, que receberam o EA. De cada cérebro, foram coletadas amostras homotípicas e semi-seriadas em cortes frontais das áreas estabelecidas para o estudo. Os resultados mostraram que as fêmeas tratadas com cipionato de testosterona apresentaram uma redução em todas as regiões analisadas já as fêmeas tratadas com Stanozolol mostraram uma diminuição em algumas áreas do hipocampo. Em relação aos animais machos, o stanozolol levou a uma diminuição na densidade neuronal em uma região do hipocampo. Estes dados nos permitem concluir que doses supra fisiológicas de esteroides utilizadas neste estudo podem causar danos consideráveis ao tecido nervoso com comprometimento ultraestrutural e consequentemente comportamental. Essas alterações podem interferir na perda de rendimento físico e no desempenho de atletas e não atletas e podem causar danos irreparáveis a indivíduos que fazem uso irresponsável destes EA.(AU)

Effects of Mlx-8, a phospholipase A2 from Brazilian coralsnake Micrurus lemniscatus venom, on muscarinic acetylcholine receptors in rat hippocampus

Abstract Background: Here, we described the presence of a neurotoxin with phospholipase A2 activity isolated from Micrurus lemniscatus venom (Mlx-8) with affinity for muscarinic acetylcholine receptors (mAChRs). Methods: The purification, molecular mass determination, partial amino acid sequencing, phospholipase A2 activity determination, inhibition of the binding of the selective muscarinic ligand [3H]QNB and inhibition of the total [3H]inositol phosphate accumulation in rat hippocampus of the Mlx-8 were determined. Results: Thirty-one fractions were collected from HPLC chromatography, and the Mlx-8 toxin was used in this work. The molecular mass of Mlx-8 is 13.628 Da. Edman degradation yielded the following sequence: NLYQFKNMIQCTNTRSWL-DFADYG-CYCGRGGSGT. The Mlx-8 had phospholipase A2 enzymatic activity. The pKi values were determined for Mlx-8 toxin and the M1 selective muscarinic antagonist pirenzepine in hippocampus membranes via [3H]QNB competition binding assays. The pKi values obtained from the analysis of Mlx-8 and pirenzepine displacement curves were 7.32 ± 0.15, n = 4 and 5.84 ± 0.18, n = 4, respectively. These results indicate that Mlx-8 has affinity for mAChRs. There was no effect on the inhibition ability of the [3H]QNB binding in hippocampus membranes when 1 µM Mlx-8 was incubated with 200 µM DEDA, an inhibitor of phospholipase A2. This suggests that the inhibition of the phospholipase A2 activity of the venom did not alter its ability to bind to displace [3H]QNB binding. In addition, the Mlx-8 toxin caused a blockade of 43.31 ± 8.86%, n = 3 and 97.42 ± 2.02%, n = 3 for 0.1 and 1 µM Mlx-8, respectively, on the total [3H]inositol phosphate content induced by 10 µM carbachol. This suggests that Mlx-8 inhibits the intracellular signaling pathway linked to activation of mAChRs in hippocampus. Conclusion: The results of the present work show, for the first time, that muscarinic receptors are also affected by the Mlx-8...(AU)

Anabolic steroids and their effects of on neuronal density in cortical areas and hippocampus of mice

Abstract Anabolic substances have been increasingly used by bodybuilders and athletes with the goal of improving performance and aesthetics. However, this practice has caused some concern to physicians and researchers because of unknowledge of consequences that the indiscriminate and illicit use of these substances can cause. Thus, this study analyzed the effects of two commercially available anabolic steroids (AS), Winstrol Depot® (Stanozolol) and Deposteron® (Testosterone Cypionate), in the neuronal density of limbic, motor and sensory regions on the cerebral cortex and in CA1, CA2, CA3 regions of the hippocampus. A total of 60 Swiss mice were used (30 males and 30 females), separated into three groups: control and two experimental groups, which received the AAS. From each brain, homotypic and semi-serial samples were taken in frontal sections from areas established for the study. The results showed that females treated with testosterone cypionate presented a reduction in all regions tested and the ones treated with Stanozolol showed a decrease in some hippocampal areas. Regarding male animals, stanozolol led to a decrease in neuron number in one hippocampal region. These data allow us to conclude that supra-physiological doses of steroids used in this study, can cause considerable damage to nervous tissue with ultrastructural and consequently behavioral impairment. These changes could interfere with the loss of physical yield and performance of athletes and non-athletes and may cause irreparable damage to individuals making irresponsible use of anabolic steroids.

Resumo As substancias anabólicas tem sido cada vez mais utilizadas por fisiculturistas e atletas com o objetivo de melhorar o desempenho e a estética. No entanto, essa prática tem causado algumas preocupações aos médicos e pesquisadores, devido ao desconhecimento das consequencias que o uso indiscriminado e ilícito dessas substâncias podem causar. Diante disso, este estudo analisou os efeitos de dois esteroides anabolizantes (EA) comercialmente disponíveis, Winstrol Depot® (Stanozolol) e Deposteron® (cipionato de testosterona), na densidade neuronal das regiões corticais límbica, motora e sensitive bem como das áreas CA1, CA2, CA3 do hipocampo. Foram utilizados 60 camundongos Swiss (30 machos e 30 fêmeas), separados em três grupos: controle e dois grupos experimentais, que receberam o EA. De cada cérebro, foram coletadas amostras homotípicas e semi-seriadas em cortes frontais das áreas estabelecidas para o estudo. Os resultados mostraram que as fêmeas tratadas com cipionato de testosterona apresentaram uma redução em todas as regiões analisadas já as fêmeas tratadas com Stanozolol mostraram uma diminuição em algumas áreas do hipocampo. Em relação aos animais machos, o stanozolol levou a uma diminuição na densidade neuronal em uma região do hipocampo. Estes dados nos permitem concluir que doses supra fisiológicas de esteroides utilizadas neste estudo podem causar danos consideráveis ao tecido nervoso com comprometimento ultraestrutural e consequentemente comportamental. Essas alterações podem interferir na perda de rendimento físico e no desempenho de atletas e não atletas e podem causar danos irreparáveis a indivíduos que fazem uso irresponsável destes EA.

Effects of Mlx-8, a phospholipase A2 from Brazilian coralsnake Micrurus lemniscatus venom, on muscarinic acetylcholine receptors in rat hippocampus

Here, we described the presence of a neurotoxin with phospholipase A2 activity isolated from Micrurus lemniscatus venom (Mlx-8) with affinity for muscarinic acetylcholine receptors (mAChRs). Methods: The purification, molecular mass determination, partial amino acid sequencing, phospholipase A2 activity determination, inhibition of the binding of the selective muscarinic ligand [3H]QNB and inhibition of the total [3H]inositol phosphate accumulation in rat hippocampus of the Mlx-8 were determined. Results: Thirty-one fractions were collected from HPLC chromatography, and the Mlx-8 toxin was used in this work. The molecular mass of Mlx-8 is 13.628 Da. Edman degradation yielded the following sequence: NLYQFKNMIQCTNTRSWL-DFADYG-CYCGRGGSGT. The Mlx-8 had phospholipase A2 enzymatic activity. The pKi values were determined for Mlx-8 toxin and the M1 selective muscarinic antagonist pirenzepine in hippocampus membranes via [3H]QNB competition binding assays. The pKi values obtained from the analysis of Mlx-8 and pirenzepine displacement curves were 7.32 ± 0.15, n = 4 and 5.84 ± 0.18, n = 4, respectively. These results indicate that Mlx-8 has affinity for mAChRs. There was no effect on the inhibition ability of the [3H]QNB binding in hippocampus membranes when 1 µM Mlx-8 was incubated with 200 µM DEDA, an inhibitor of phospholipase A2. This suggests that the inhibition of the phospholipase A2 activity of the venom did not alter its ability to bind to displace [3H]QNB binding. In addition, the Mlx-8 toxin caused a blockade of 43.31 ± 8.86%, n = 3 and 97.42 ± 2.02%, n = 3 for 0.1 and 1 µM Mlx-8, respectively, on the total [3H]inositol phosphate content induced by 10 µM carbachol. This suggests that Mlx-8 inhibits the intracellular signaling pathway linked to activation of mAChRs in hippocampus. Conclusion: The results of the present work show, for the first time, that muscarinic receptors are also affected by the Mlx-8 toxin, a muscarinic ligand with phospholipase A2 characteristics, obtained from the venom of the Elapidae snake Micrurus lemniscatus, since this toxin was able to compete with muscarinic ligand [3H]QNB in hippocampus of rats. In addition, Mlx-8 also blocked the accumulation of total [3H]inositol phosphate induced by muscarinic agonist carbachol. Thus, Mlx-8 may be a new pharmacological tool for examining muscarinic cholinergic function.(AU)

The microanatomy of the central nervous system and brain of the Indo-Pacific seahorse, Hippocampus barbouri, during development

The central nervous system (CNS) of Teleostei is a complex system of self-governance and its morphology is reflected in the physiological and reproductive behaviors. The Indo-Pacific seahorse, Hippocampus barbouri Jordan & Richard son, 1908, is a new candidate species for aquaculture in Thailand. In this study, we investigated the brain morphology of H. barbouri across various developmental windows. Light microscopic observations of adult brains revealed a large optic tectum in the mesencephalon, whereas the cerebral hemispheres and the cerebellum are of medium size. The detailed brain structures were generally similar to those of other teleosts; however, only five distinct layers were present in the optic tectum, including the stratum marginale, stratum opticum, stratum album central, stratum griseum central, and stratum periventriculae, versus six layers observed in other fish. One day after birth (1 DAB) the brain was a packed structure without any clear sub-structures. The number of capillaries in the optic tectum began to increase at 6 DAB, and at 14 DAB several features, including small blood vessels in the optic tectum and Purkinje cells, became noticeable. By 35 DAB, the optic tectum became highly vascularized and included five layers. Additionally, large Purkinje cells were developed in the cerebellum. Based on the brain development pattern, we speculate that the predatory ability of this fish starts to develop from 6 to 14 days after birth.

The microanatomy of the central nervous system and brain of the Indo-Pacific seahorse, Hippocampus barbouri, during development

The central nervous system (CNS) of Teleostei is a complex system of self-governance and its morphology is reflected in the physiological and reproductive behaviors. The Indo-Pacific seahorse, Hippocampus barbouri Jordan & Richard son, 1908, is a new candidate species for aquaculture in Thailand. In this study, we investigated the brain morphology of H. barbouri across various developmental windows. Light microscopic observations of adult brains revealed a large optic tectum in the mesencephalon, whereas the cerebral hemispheres and the cerebellum are of medium size. The detailed brain structures were generally similar to those of other teleosts; however, only five distinct layers were present in the optic tectum, including the stratum marginale, stratum opticum, stratum album central, stratum griseum central, and stratum periventriculae, versus six layers observed in other fish. One day after birth (1 DAB) the brain was a packed structure without any clear sub-structures. The number of capillaries in the optic tectum began to increase at 6 DAB, and at 14 DAB several features, including small blood vessels in the optic tectum and Purkinje cells, became noticeable. By 35 DAB, the optic tectum became highly vascularized and included five layers. Additionally, large Purkinje cells were developed in the cerebellum. Based on the brain development pattern, we speculate that the predatory ability of this fish starts to develop from 6 to 14 days after birth.(AU)

Systematization, distribution, and territories of the caudal cerebral artery on the surface of the brain in nutria (Myocastor coypus) / Sistematização, distribuição e territórios da artéria cerebral caudal na superfície do cérebro em Nutria (Myocastor coypus)

The nutria (Myocastor coypus) is a medium-sized, semi-aquatic rodent valued by the skin and meat industry. This study aimed to describe and systematize the caudal cerebral artery on the brain surface in nutria, establishing a standard model and its main variations in this species. The thirty animals used were euthanized according to animal welfare rules. The vessels were filled with latex stained with red pigment and the samples were fixed in formaldehyde. In nutria, the brain was vascularized by the vertebral basilar system. The terminal branches of the basilar artery originated the rostral cerebellar, caudal cerebral, rostral choroidal and middle cerebral arteries, and its terminal branch, the rostral cerebral artery. The terminal branch of the basilar artery projected the caudal cerebral artery, which is usually a single medium-caliber vessel, into the transverse fissure of the brain. The caudal cerebral artery was presented as a single (66.7% of the cases to the right and 76.7% to the left) and double vessel (33.3% of the cases to the right and 23.3% to the left). It originated the rostral mesencephalic artery, the proximal component, and the caudal inter-hemispheric artery. The terminal branches of the rostral and caudal tectal mesencephalic arteries formed a typical anastomotic network. The caudal inter-hemispheric artery emitted central branches, the caudal choroidal artery, hemispherical occipital arteries, rostral tectal mesencephalic branches and distal components, and anastomosed "in osculum" with the terminal branches of the rostral inter-hemispheric artery. The caudal choroidal artery anastomosed with the rostral choroidal artery, where it branched out on the thalamic mass, vascularizing all diencephalic structures and the hippocampus. The caudal cerebral artery and its terminal branches anastomosed with the terminal branches of the rostral and middle cerebral arteries in a restricted region of the caudal pole of the cerebral hemisphere. The vascularization area of the caudal cerebral artery and its central branches in the paleopallial of the piriform lobe is extremely restricted, caudomedially.(AU)

A nutria (Myocastor coypus) é um roedor semi-aquático de tamanho mediano, apreciado na indústria de peles e carne. Este trabalho tem por objetivo descrever e sistematizar a artéria cerebral caudal na superfície do cérebro em nutria, estabelecendo um modelo padrão e suas principais variações e territórios nesta espécie. Os trinta animais utilizados foram eutanasiados segundo as regras de bem-estar animal, os vasos foram preenchidos com látex, corado em vermelho e as peças foram fixadas em formoldeído. O cérebro foi vascularizado exclusivamente pelo sistema vértebro-basilar. Os ramos terminais da artéria basilar originaram as artérias cerebelar rostral, cerebral caudal, corióidea rostral, cerebral média e seu ramo terminal, a artéria cerebral rostral. O ramo terminal da artéria basilar lançou a artéria cerebral caudal, um vaso normalmente único, de médio calibre, para o interior da fissura transversa do cérebro. A artéria cerebral caudal foi um vaso único em 66,7% à direita e em 76,7% à esquerda e mostrou-se dupla em 33,3% à direita e em 23,3% à esquerda. Ela lançou a artéria tectal mesencefálica rostral, componente proximal e a artéria inter-hemisférica caudal. Os ramos terminais das artérias tectais mesencefálicas, rostral e caudal, formavam uma rede anastomótica típica. A artéria inter-hemisférica caudal lançou ramos centrais, a artéria corióidea caudal, as artérias hemisféricas occipitais, os ramos tectais mesencefálicos rostrais, componentes distais e anastomosou-se "em ósculo" com o ramo terminal da artéria inter-hemisférica rostral. A artéria corióidea caudal anastomosava-se com a artéria corióidea rostral, onde ramificavam-se sobre a massa talâmica, vascularizando todas as estruturas do diencéfalo e hipocampo. A artéria cerebral caudal com seus ramos terminais apresenta anastomoses com os ramos terminais das artérias cerebrais rostral e média em uma região restrita do pólo caudal do hemisfério cerebral. A área de vascularização da artéria cerebral caudal com seus ramos centrais no páleo-palio do lobo piriforme é extremamente restrita, caudo-medialmente ao mesmo.(AU)

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...(AU)