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.

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

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.

Oral pilocarpine for the treatment of dry eye in patients with Sjögren's syndrome / Pilocarpina oral no tratamento do olho seco de pacientes com síndrome de Sjögren

ABSTRACT Purpose: To evaluate the efficacy of oral pilocarpine (20 mg daily) for the treatment of dry eye in patients with Sjogren's Syndrome. The frequency of side effects reported during the treatment was also investigated. Methods: In this placebo-controlled crossover study, 32 patients with Sjögren's syndrome were enrolled to receive either oral pilocarpine or placebo for 10 weeks. Following a 2-week washout period, the treatment was inverted for each patient for the same duration. Assessments included the quality of life National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25), dry eye specific questionnaire Ocular Surface Disease Index, non-invasive breakup time, invasive breakup time with fluorescein, corneal and conjunctival staining patterns with the use of fluorescein and rose bengal staining, Schirmer's test, and tear ferning test. Results: According to the NEI-VFQ-25, there was statistically significant improvement in the quality of life following oral pilocarpine. Similar results were observed for ocular discomfort, as determined by the Ocular Surface Disease Index. All clinical tests showed favorable and statistically significant results following the use of oral pilocarpine. Regarding the analysis of tear samples, there was an improvement in the quality of tear film. This was evidenced by the modification of the patterns observed in the tear ferning test. Side effects were reported by 96.8% and 56.2% of the patients who received pilocarpine and placebo, respectively. Sweating was the most frequently reported side effect (67.74% versus 11.11%, respectively). Conclusions: Although the treatment was associated with a high frequency of side effects, oral pilocarpine (20 mg daily) was able to relieve discomfort related to dry eyes in patients with Sjögren's syndrome and induce favorable structural changes in the tear film.

RESUMO Objetivos: Avaliar o alívio de sintomas e sinais relacionados à secura ocular e na qualidade de vida de pacientes com síndrome de Sjögren tratados com o uso oral de pilocarpina na dose diária de 20mg. A frequência dos efeitos colaterais relatados com o tratamento também foi estudada. Métodos: Trata-se de estudo cruzado e placebo-controlado, que envolveu 32 pacientes com síndrome de Sjögren em uso de pilocarpina oral ou placebo, por dez semanas. Após duas semanas sem medicações, houve a inversão dos tratamentos para cada paciente, por mais dez semanas. As avaliações foram feitas por meio do questionário de qualidade de vida NEI-VFQ-25, questionário olho seco específico Ocular Surface Disease Index, tempo de ruptura do filme lacrimal não invasivo, tempo de ruptura do filme lacrimal com fluoresceína, avaliação da superfície ocular com os corantes fluoresceína e rosa Bengala, teste de Schirmer e teste de cristalização do filme lacrimal. Resultados: Houve melhora estatisticamente significante na qualidade de vida medida pelo questionário NEI-VFQ-25 e no desconforto ocular avaliado pelo Ocular Surface Disease Index, após o tratamento. Todos os testes clínicos sofreram influência favorável e estatisticamente significante durante a fase de tratamento com pilocarpina oral. Em relação à análise de amostras de lágrimas, ocorreram alterações estruturais indicando melhora no padrão de cristalização do filme lacrimal. Os efeitos colaterais com o uso de pilocarpina foram relatados por 96,8% dos pacientes com a pilocarpina e 56,2% com placebo. Sudorese foi o efeito colateral mais frequentemente relatado (pilocarpina = 67,74%; placebo = 11,11%). Conclusões: O uso oral de pilocarpina na dose diária de 20mg foi capaz de aliviar as queixas de desconforto relacionadas ao ressecamento ocular em pacientes com síndrome de Sjögren, produzir impacto positivo na qualidade de vida dos pacientes e induzir mudanças estruturais favoráveis no filme lacrimal, embora os efeitos colaterais relatados tenham ocorrido com alta frequência.

The effects of pharmacological accommodation and cycloplegia on axial length and choroidal thickness / Efeitos da acomodação farmacológica e da cicloplegia no comprimento axial e na espessura da coroide

ABSTRACT Purpose: To investigate the effects of pharmacological accommodation and cycloplegia on ocular measurements. Methods: Thirty-three healthy subjects [mean (±SD) age, 32.97 (±5.21) years] volunteered to participate in the study. Measurement of the axial length, macular and choroidal thickness, refractive error, and corneal topography, as well as anterior segment imaging, were performed. After these procedures, pharmacological accommodation was induced by applying pilocarpine eye drops (pilocarpine hydrochloride 2%), and the measurements were repeated. The measurements were repeated again after full cycloplegia was induced using cyclopentolate eye drops (cyclopentolate hydrochloride 1%). The correlations between the measurements were evaluated. Results: A significant increase in subfoveal choroidal thickness after applying 2% pilocarpine was identified (without the drops, 319.36 ± 90.08 µm; with pilocarpine instillation, 341.60 ± 99.19 µm; with cyclopentolate instillation, 318.36 ± 103.0 µm; p<0.001). A significant increase in the axial length was also detected (without the drops, 23.26 ± 0.83 mm; with pilocarpine instillation, 23.29 ± 0.84 mm; with cyclopentolate instillation, 23.27 ± 0.84 mm; p=0.003). Comparing pharmacological accommodation and cycloplegia revealed a significant difference in central macular thickness (with pilocarpine instillation, 262.27 ± 19.34 µm; with cyclopentolate instillation, 265.93 ± 17.91 µm; p=0.016). Pilocarpine-related miosis (p<0.001) and myopic shift (p<0.001) were more severe in blue eyes vs. brown eyes. Conclusion: Pharmacological accommodation may change ocular measurements, such as choroidal thickness and axial length. This condition should be considered when performing ocular measurements, such as intraocular lens power calculations.(AU)

RESUMO Objetivo: Investigar os efeitos da acomodação farmacológica e da cicloplegia nas medições oculares. Métodos: participaram do estudo 33 voluntários saudáveis (média de idade [± DP], 32,97 anos [± 5,21 anos]). Foram medidos o comprimento axial, a espessura macular e coroidal e o erro refrativo, bem como realizados exames de imagem da topografia corneana e do segmento anterior. Em seguida, foi induzida a acomodação farmacológica aplicando-se colírio de pilocarpina (cloridrato de pilocarpina a 2%) e as medições foram repetidas nos participantes. As mesmas medições foram repetidas depois de induzir a cicloplegia completa com colírio de ciclopentolato (cloridrato de ciclopentolato a 1%) e foram avaliadas as correlações entre as medidas. Resultados: Identificou-se aumento significativo da espessura coroidal subfoveal com o uso da pilocarpina a 2% (sem colírio, 319,36 ± 90,08 µm; com a instilação de pilocarpina, 341,60 ± 99,19 µm; com a instilação de ciclopentolato, 318,36 ± 103,0 µm; p<0,001). Detectou-se também aumento significativo do comprimento axial (sem colírio, 23,26 ± 0,83 mm; com a instilação de pilocarpina, 23,29 ± 0,84 mm; com a instilação de ciclopentolato, 23,27 ± 0,84 mm; p=0,003). Ao se comparar a acomodação farmacológica e a cicloplegia, houve diferença significativa na espessura macular central (com a instilação de pilocarpina, 262,27 ± 19,34 µm; com a instilação de ciclopentolato, 265,93 ± 17,91 µm; p=0,016). Observou-se que a miose associada à pilocarpina (p<0,001) e o desvio miópico (p<0,001) foram mais severos nos olhos azuis que nos castanhos. Conclusão: A acomodação farmacológica pode alterar medidas oculares como a espessura da coroide e o comprimento axial. Essa possibilidade deve ser levada em consideração ao se efetuarem medições oculares, tais como cálculos de potência de lentes intraoculares.(AU)

Consistency between anticholinergic burden scales in patients with Sjögren's syndrome / Consistencia entre escalas de carga anticolinérgica en pacientes con síndrome de Sjögren

ABSTRACT Introduction: Sjögren's syndrome is the second most frequent autoimmune rheumatic disease and is characterized by exocrine gland involvement manifesting as sicca symptoms. The objective of this study was to estimate the degree of agreement between three anticholinergic burden scales related to the prescriptions of patients diagnosed with Sjögren's syndrome in Colombia. Materials and methods: An analytical concordance study was conducted. The weighted kappa coefficient with quadratic weights was used to identify consistency between the Anticholinergic Drug Scale (ADS), Anticholinergic Cognitive Burden Scale (ACB) and Anticholinergic Risk Scale (ARS), which address the prescriptions used for 3 months by patients with Sjögren s syndrome, in a population database. Results: A total of 15,696 patients with Sjögren's syndrome were identified, with a mean age of 65.4 ± 13.9 years, and 74.2% were women. A total of 94.1% of the patients received at least one topical lubricant, with carboxymethyl cellulose being the most commonly prescribed (22.9%), while oral pilocarpine was prescribed to 3.5% of patients. The ACB was the tool identified more antimuscarinic prescriptions (37.5%), followed by the ADS (35.3%) and ARS (25.2%). The greatest degree of agreement was found between the ADS and ACB (kappa 0.6520; confidence interval (CI): 0.6393-0.6648). Conclusions: Except for the ADS and ACB, little agreement was found between the three scales gauging the anticholinergic burden. Additional studies are needed to determine how these differences can impact the clinical outcomes of patients.

RESUMEN Introducción: El síndrome de Sjögren es la segunda enfermedad reumática autoinmune más frecuente, caracterizada especialmente por el compromiso de glándulas exocrinas manifestándose con síntomas sicca. El objetivo fue estimar el grado de acuerdo de 3 escalas de carga anticolinérgica en las prescripciones de pacientes con diagnóstico de síndrome de Sjögren en Colombia. Materiales y métodos: Estudio analítico de concordancia. Se empleó el coeficiente Kappa con ponderación cuadrática para identificar la consistencia entre los instrumentos Anticholin-ergic Drug Scale (ADS), Anticholinergic Cognitive Burden Scale (ACB) y Anticholinergic Risk Scale (ARS) de las prescripciones utilizadas durante 3 meses por pacientes con síndrome de Sjögren, a partir de una base de datos poblacional. Resultados: Se identificaron 15.696 pacientes con síndrome de Sjögren, con una edad media de 65,4 ±13,9 anos y el 74,2% de mujeres. El 94,1% recibieron por lo menos un lubricante tópico siendo el más prescrito la carboximetilcelulosa (22,9%), mientras que la pilocarpina oral se formuló en el 3,5% de ellos. La escala ACB fue la herramienta que más prescripciones anti-muscarínicas identificó (37,5%) seguida de la ADS (35,3%) y ARS (25,2%). El mayor grado de acuerdo se presentó entre las escalas ADS-ACB (Kappa: 0,6520; IC: 0,6393-0,6648). Conclusiones: Con excepción de las escalas ADS-ACB, hubo poco acuerdo al comparar las 3 escalas de carga anticolinérgica. Se requieren estudios adicionales para determinar cómo estas diferencias pueden impactar en la validez de los desenlaces clínicos de los pacientes.

A visão em pseudofácicos com ametropia residual após instilação de pilocarpina a 2% / Pseudophakic Vision with residual ametropia after pilocarpine 2% instillation

Resumo Objetivo: Avaliar modificações de acuidade visual, refração, campo visual e diâmetro pupilar, em pacientes pseudofácicos, após a instilação de pilocarpina a 2%. Métodos: Ensaio clínico, controlado, mascarado e randomizado realizado entre maio de 2015 e setembro de 2016 no Hospital Universitário Gaffrée e Guinle, RJ, Brasil. Quarenta pacientes divididos em 2 grupos foram acompanhados em pós-operatório de facectomia com implante de LIO. No grupo de casos houve aplicação de uma gota de pilocarpina a 2%, no grupo controle, uma gota de lubrificante no olho operado. Foram avaliadas antes e 1 hora após a instilação do colirio: a acuidade visual para longe e perto com e sem correção; a refração; o diâmetro pupilar e o campo visual. Resultados: No grupo de casos, a acuidade visual s/c para longe aumentou de 0,33 para 0,57 (p = 0,0001) e para perto melhorou também, 13 pacientes (59,09%) possuíam acuidade visual de J1 ou J2 antes da instilação e depois o número aumentou para 18 ou 81,81% (p = 0,0054). O diâmetro pupilar reduziu de 2,00mm para 1,85mm (p < 0,0001). Não houve alteração do campo visual central. No grupo controle, não houve variação estatisticamente ou clinicamente significativa de qualquer um dos parâmetros medidos. Conclusão: A administração tópica de uma gota de pilocarpina a 2% melhorou a visão de pacientes pseudofácicos com ametropia residual para longe e para perto. Estudos de dose-efetividade adicionais podem indicar melhores concentrações e posologias para alcançar maiores melhoras de acuidade visual.

Abstract Objective: Evaluate the visual acuity, refraction, visual field changes and pupillary diameter in pseudophakic patients after instillation of 2% pilocarpine eye drops. Methods: Controlled, masked and randomized clinical trial carried out between May, 2015 and September, 2016 at the Gaffrée and Guinle University Hospital, RJ, Brazil. Forty patients, divided into 2 groups, were followed up in the postoperative period of a facectomy with intraocular lens implant. The patients in the group of cases were submitted to a drop of 2% pilocarpine and those of the control group to a drop of lubricant in the operated eye. Before eye drop instillation nd one hour after it, the authors evaluated: visual acuity for distance and near; refraction; pupillary diameter and visual field. Results: In case group visual acuity increased from 0.33 to 0.57 for far (p = 0.0001)and also increased for near, 13 patients (59.09%) had visual acuity of J1 or J2 before instillation and 18 or 81.81% after it (p = 0.0054). The median pupillary diameters raised from 2.00 mms to 1.85 mm(p <0.0001). Central visual fields did not have significant alteration. In the control group, there were no statistically or clinically significant changes in any of the measured parameters. Conclusion: Topical administration of a 2% pilocarpine eye drop was effective to improve pseudophakic patients vision with residual ametropia for far and near. Additional dose-effectiveness studies may indicate better concentrations and dosages to achieve greater improvements in visual acuity.

Pupillometry in Chagas disease / Pupilometria na doença de Chagas

ABSTRACT Purpose: We investigated parasympathetic innervation abnormalities of the iris sphincter and ciliary muscles in chronic Chagas disease by measuring pupillary diameter and intraocular pressure. Methods: A group of 80 patients with Chagas disease was compared with 76 healthy individuals without chagasic infection. The following procedures were performed: pupillometry, hypersensitivity test to pilocarpine 0.125%, intraocular pressure measurement (IOP), basal pupil diameter (BPD), absolute pupillary constriction amplitude (ACA), relative pupillary constriction amplitude (RCA) and the presence of anisocoria. Results: The prevalence of anisocoria was higher in chagasic patients (p<0.01). These patients had mean basal pupillary diameter, mean photopic pupillary diameter and mean value of absolute pupillary constriction amplitude significantly lower than non-chagasic ones (p<0.01, mean difference -0.50mm), (p=0.02, mean difference -0.20mm), (p<0.01, mean difference -0.29mm), respectively. The relative pupillary constriction amplitude did not differ between the two groups (p=0.39, mean difference -1.15%). There was hypersensitivity to dilute pilocarpine in 8 (10%) of the chagasic patients in the right eye and in 2 (2.5%) in the left eye and in 1 (1.25%) in both eyes. The mean value of intraocular pressure had a marginal statistical significance between the two groups (p=0.06, mean difference -0.91mmHg). Conclusions: Patients with chagasic infection may exhibit ocular parasympathetic dysfunction, demonstrable by pupillometry and the dilute pilocarpine hypersensitivity test.

RESUMO Introdução: Investigaram-se anormalidades da inervação parassimpática dos músculos esfíncter da íris e ciliar na doença de Chagas crônica, através de medidas pupilares e da pressão intraocular. Métodos: Foram estudados dois grupos, um com 80 chagásicos e outro com 76 indivíduos saudáveis sem infecção chagásica. Foram realizados os seguintes procedimentos: pupilometria, teste de hipersensibilidade à pilocarpina a 0,125%, medida da pressão intraocular (PIO), diâmetro basal da pupila (DBP), amplitude de constrição pupilar absoluta (ACA), amplitude de constrição pupilar relativa (ACR), e presença de anisocoria. Resultados: A prevalência de anisocoria foi maior nos chagásicos (p<0,01). Estes pacientes apresentaram diâmetro basal pupilar médio, diâmetro fotópico médio e valor médio da amplitude de constrição pupilar absoluta, significativamente menores que os não chagásicos, (p<0,01, diferença de média -0,50mm), (p=0.02, diferença de média -0,20mm), (p<0,01, diferença de média -0,29mm), respectivamente. A amplitude de constrição pupilar relativa não diferiu entre os dois grupos (p=0,39, diferença de média -1,15%). Houve hipersensibilidade à pilocarpina diluída em 8 (10%) chagásicos no olho direito em 2 (2,5%) no olho esquerdo e em 1 (1,25%) em ambos os olhos. O valor médio da pressão intraocular teve significância marginal entre os dois grupos (p=0,06, diferença de média -0,91mmHg). Conclusões: Pacientes com infecção chagásica podem apresentar disfunção parassimpática ocular, demonstrável pela pupilometria e pelo teste de hipersensibilidade à pilocarpina diluída.

Promotion of epileptic seizure through up-regulation of matrix metalloproteinase-9 by apoptosis signal-regulating kinase 1 in lithium-pilocarpine epilepsy model / 实用医学杂志

Objective To investigate the alteration of apoptosis signal-regulating kinase 1(ASK1)phos-phorylation and matrix metalloproteinase-9(MMP-9)expression after temporal lobe epilepsy(TLE),and to explore the role and mechanism of ASK1-MMP-9 signal in TLE. Methods The lithium-pilocarpine epilepsy model was established. The ASK1 phosphorylation and MMP-9 expression were detected by western blot and immunofluo-rescence histochemistry. Using ASK1 inhibitors,the regulating effects of ASK1 on MMP-9 expression were deter-mined. And the role of ASK1 and MMP-9 in epileptic seizure was explored by inhibition of ASK1 and MMP-9. Results The ASK1 phosphorylation and MMP-9 expression increased in hippocampus in epilepsy model ,exhibiting a temporal correlation with epileptic seizure. Inhibition of ASK1 activation reduced the expression of MMP-9. And the level of epileptic seizure was significantly reduced by the inhibition of ASK1 and MMP-9. Conclusions The hippocampal ASK1 phosphorylation and MMP-9 expression distinctly increase in TLE model ,and the MMP-9 expression is induced by ASK1 activation. The ASK1-MMP-9 signal plays an important role in TLE.

Expression of nucleotide-binding oligomerization domain-like receptor protein 3 inflammatory corpuscles in epileptic model and the effect of melatonin on its expression / 中华实用儿科临床杂志

Objective To observe the expression of nucleotide-binding oligomerization domain-like recep-tor protein 3(NLRP3)inflammasome in epilepsy model,and to explore the neuroprotective effect of melatonin. Methods SD rats aged 21-30 d were randomly divided into the control group(48 rats),the epilepsy group(48 rats)and the melatonin group(48 rats),and each group was subdivided into 4 subgroups according to the time points of 24 h,48 h, 72 h,and 7 d,with 12 SD rats in each subgroup. Fluorescence quantitative polymerase chain reaction and immunohisto-chemical technique were used to analyze the expressions of NLRP3,Caspase-1 and interleukin(IL)-1β in hippocam-pus areas of rats at different points of time after seizures were induced,and their behavior changes were observed. Results The number of NLRP3-positive cells in the epileptic group increased,and reached the peak at 72 h. At 24 h,48 h,72 h,7 d,the number of NLRP3-positive cells in the epilepsy group(14. 20 ± 1. 64,23. 60 ± 1. 14,31. 20 ± 1. 30,25. 40 ± 2. 07)was significantly increased compared with those of the melatonin group(10. 60 ± 0. 89,17. 80 ± 1. 48,24. 00 ± 0. 71,20. 20 ± 1. 92)and the control group(2. 60 ± 0. 89,2. 40 ± 1. 14,2. 40 ± 1. 14,2. 40 ± 0. 55),and the differences were significant(F＝122. 977,375. 125,962. 743,262. 916,all P＜0. 05). The NLRP3 mRNA relative expressions in the epilepsy group (2. 57 ± 0. 12,3. 34 ± 0. 10,4. 84 ± 0. 19,3. 55 ± 0. 13)were significantly increased compared with those of the melatonin group (2. 03 ± 0. 08,2. 71 ± 0. 08,4. 03 ± 0. 14,2. 48 ± 0. 18)and the control group(1. 07 ± 0. 13,1. 08 ± 0. 15,1. 08 ± 0. 23,1. 07 ± 0. 18),and the differences were significant (F ＝422. 386, 1 154. 957,1 132. 112,512. 149,all P ＜0. 05);the Caspase -1 mRNA relative expressions in the epilepsy group (2. 47 ± 0. 07,3. 05 ± 0. 15,4. 39 ± 0. 18,3. 14 ± 0. 11)were significantly increased compared with those of melatonin group(1. 85 ± 0. 07,2. 49 ± 0. 08,3. 60 ± 0. 12,2. 15 ± 0. 12)and the control group (0. 98 ± 0. 25,0. 99 ± 0. 15,0. 98 ± 0. 23,0. 99 ± 0. 18),and the differences were significant(F ＝620. 099,580. 796,1 125. 225,645. 082,all P ＜0. 05);the IL-1β mRNA relative expressions in epilepsy group (2. 32 ± 0. 15,2. 90 ± 0. 18,4. 18 ± 0. 16,2. 74 ± 0. 07)were significantly increased compared with those of the melatonin group (1. 78 ± 0. 09,2. 35 ± 0. 11,3. 24 ± 0. 13,1. 78 ± 0. 16)and the control group(0. 97 ± 0. 13,0. 99 ± 0. 15,0. 97 ± 0. 23,0. 97 ± 0. 18),and the differences were significant(F＝267. 952,398. 767,1 140. 384,438. 962,all P ＜0. 05). Conclusions The NLRP3 inflamma-somes are activated in rat hippocampus with epilepsy induced by lithium-pilocarpine. NLRP3 inflammasome mediated inflammatory response probably involved in the pathogenesis of epilepsy. The melatonin may play a neuroprotective role by inhibiting expression of NLRP3 inflammasome.

Change in hippocampal voltage-gated Ⅰ sodium channel of Lithium chloride-Pilocarpine epileptic rat model / 中华实用儿科临床杂志

Objective To research the changes in hippocampal voltage-gated sodium channel of Lithium chloride-Pilocarpine epileptic rat models,including Ⅰ sodium channel α subunit protein (Nav1.1),mRNA of Ⅰ sodium channel alpha subunit protein gene and function of sodium channel.Methods Epileptic rat models of Lithium chloride-Pilocarpine were established.Nav1.1 expression in the hippocampus of experimental rats was detected by immunohistochemical staining method,and the changes in voltage-gated sodium channel function (the current-voltage curves,activation and inactivation curves and the recovery curve) of hippocampus nerve cells were detected by whole cell patch-clamp technique.Results (1) The Lithium chloride-Pilocarpine rat models were successfully reproduced.Three stages of behavior (acute,latent and chronic) of rat models were observed.The blank control group was free of seizure.(2) Immunohistochemistry results:neurons in CA1 and DG regions of hippocampal of epileptic rats were normal,and there was no obvious change in the expression of Nav1.1.In CA3 area,the degeneration and necrosis of neurons were obvious.Staining of Nav1.1 became superficial and even disappeared in these areas,but the normal tissues were enhanced around degenerative and necrotic neurons.Compared with the blank control group,the expression of Nav1.1 in the model group was higher(0.235 ±0.008 vs.O.210 ±0.002),and there was statistically significant difference (t'=-7.426,P ＜ 0.05).(3) The whole-cell patch-clamp technique showed that the sodium current density of the model group increased significantly compared with that of the blank group [(-319.70 ± 28.24) pA/pF vs.(-229.06 ± 26.01) pA/pF,t =8.178,P ＜ 0.05],the threshold value of activation curve decreased (4.15 ± 0.80 vs.4.50 ±0.85,t =11.020,P ＜ 0.05),the threshold value of inactivation curve increased (7.47 ± 0.53 vs.6.24 ±0.31,t =6.940,P ＜ 0.05),and the recovery time after inactivation shortened [(1.36 ± 0.15) ms vs.(1.86 ± 0.21)ms,t =6.712,P ＜ 0.05],and there were all statistically significant differences.Conclusion Repeated seizures can lead to increase Nav1.1 compensatory expression of,and significantly increase sodium channel current density,while the threshold value of activation curve decreases,the threshold value of inactivation curve rises,and the recovery time after inactivation is shortened,which eventually leads to increased neuron excitability and is more likely to cause seizures.

Terapia farmacológica y avances terapéuticos en xerostomía e hiposalivación / Pharmacological therapy and therapeutic advancesin xerostomy and hyposalivation

La saliva juega un rol determinante en el mantenimiento y función de los tejidos orales, donde un decremento en los niveles del flujo salival conllevan a una hiposalivación o hiposialia, trayendo consigo una serie de características que propician un ambiente ideal para la instalaciónde microorganismos oportunistas y condiciones nocivas para la saludoral y sistémica de los pacientes. Hoy en día existen múltiples causas que conllevan a una hiposialia, entre ellas el síndrome de Sjõgren, la radioterapia de haz externo, entre otras, los cuales se han puesto a prueba mediante sialogogos farmacológicos como pilocarpina, cevimelina, betanecoly carbacolina, además de alternativas terapéuticas para revertir los signos obtenidos por la hiposalivación y mitigar los síntomas de xerostomía. El objetivo del presente es realizar una revisión de literatura sobre el tratamiento farmacológico en hiposalivación y xerostomía ensíndrome de Sjõgren y radioterapia de haz externo.

Saliva plays a determinant role in the maintenance and function of oral tissues, where a decrease in salivary flow levels leads to hyposalivation or hyposialia, bringing with it a series of characteristics that provide anideal environment for the installation of opportunistic microorganisms and conditions harmful to the oral and systemic health of patients.Today there are many causes that lead to hyposialia, including Sjögren’ssyndrome, external beam radiotherapy, among others, which have beentested by pharmacological sialogogs such as pilocarpine, cevimelin, betanecol and carbacoline, in addition to therapeutic alternativesto reverse the signs obtained by hyposalivation and to mitigate the symptoms of xerostomia. The objective of the present is to make a reviewof the literature on the pharmacological treatment in hyposalivation and xerostomia in Sjögren’s syndrome and external beam radiotherapy.

Salivary stimulation by prolonged release of pilocarpine in sjõgren’ syndrome

Introduction: Prolonged drug delivery in the oral cavity offers many advantages, such as reducing adverse effects. Pilocarpine is an FDA-approved parasympathomimetic drug for the treatment of glandular hypofunction; however, its adverse effects limit its use. Objective: To evaluate the stimulation of salivary flow by the use of pilocarpine-releasing films, as well as their effects on the symptoms of xerostomia and adverse effects in patients with Sjõgren’s syndrome (SS). Materials and methods: Hydroxypropylmethylcellulose (Methocel K4MCR) films were prepared in 1 percent acetic acid and pilocarpine was added under magnetic stirring. The pH and thickness, as well as diffusion uniformity and kinetics of drug release per cm2 were evaluated by spectrophotometry. The films were tested sublingually in 40 patients with Sjõgren’s syndrome for a period of two weeks. Changes in their salivary flow were evaluated by analyzing samples of total saliva. Additionally, patients were screened for symptoms of xerostomia and adverse effects. Results: The films had a pH of 2.91 +/- 0.035, a thickness of 0.06866 +/- 0.00152μm, and a diffusion uniformity of 91 percent per cm2. Use of the films resulted in an increase in salivary flow in both primary and secondary Sjõgren’s syndrome, but this increase was only significant in primary SS. Conclusion: Films showed optimal physicochemical properties for their administration, and proved effective in stimulating salivary flow without causing adverse effects during their administration.

Purslane protects against the reproductive toxicity of carbamazepine treatment in pilocarpine-induced epilepsy model

Objective To investigate the protective effect of purslane with carbamazepine treatment. Methods Male albino rats were modulated by pilocarpine to be epileptic. Both the normal and epileptic rats were treated with carbamazepine, purslane or carbamazepine plus purslane, with separate non-treated control groups for both normal and epileptic rats. Results The data from the current study showed amelioration in amino acids and electrolytes in the epileptic rats treated with purslane and carbamazepine, with this amelioration occurring without decreasing the fertility hormones (testosterone, dehydroepiandrosterone, luteinizing hormone and follicle stimulating hormone). Purslane treatments also prevented the increase in estradiol. The decreased epileptic hyperexcitability with purslane was evidenced by decreased glial fibrillary acidic protein and lipid peroxidation. Conclusions Natural products like purslane could be used with the highly repetitive drugs like carbamazepine to reduce or prevent its side-effects.

The clinical efficacy of combined pilocarpine eye drops in the treatment of traumatic angle recession glaucoma / 中国生化药物杂志

Objective To evaluate the clinical efficacy of combined pilocarpine eye drops in the treatment of traumatic angle recession glaucoma.MethodsThe clinical data of the patients were collected and analyzed retrospectively.The data collection time was December 2015 to December 2016.Were randomly divided into two groups according to the time of admission, the control group (38 cases) received routine treatment, the observation group (40 cases) treated with conventional therapy combined with pilocarpine eye drops.The IOP and pupil diameter of two groups were observed and compared.ResultsAfter treatment, two groups of patients with intraocular pressure were significantly decreased, compared with the group before treatment, there were statistically significant differences, and different time after the treatment were compared between the observation group were significantly lower than the control group, the difference was significant (P<0.05);after two 7d of treatment group were significantly reduced compared to the diameter of the pupil.Compared with the group before treatment, there were statistically significant differences, and different time after the treatment were compared between the observation group were significantly less than the control group, the difference was significant (P<0.05);efficiency evaluation results of two groups under different treatment, the observation group treated a total of 21 cases invalid, only 2 cases, the total effective rate was 95.00%, and there was no statistic difference between control group (P<0.05).ConclusionThe conventional therapy combined with pilocarpine eye drops on injury caused by angle recession glaucoma patients for treatment can significantly reduce intraocular pressure, miosis, improve efficiency.

Combined administration of pilocarpine and timolol eye drops for the treatment of glaucoma / 中国生化药物杂志

Objective To analyze the therapeutic effect of combined administration of pilocarpine and timolol eye drops in the treatment of glaucoma, and to summarize the rational administration of the two drugs.Methods The study was conducted by animal experiment.The study was conducted from January 2016 to February 2016.Fifty New Zealand white rabbits were randomly divided into 5 groups: pilocarpine group, timolol group, conventional administration group, pilocarpine+timolol group, timolol group and pilocarpine group.each group had 10 New Zealand white rabbits, according to different modes of administration,,the changes in intraocular pressure after 10, 20, 30, 40, 60, 90, 120, 150, 180 and 240 min after the first administration were compared.Results Intraocular pressure changes with time points after administration of different administration methods, There was no significant difference of IOP changes in pilocarpine group and timolol group single drug.pilocarpine+timolol timolol group changes with time continuing to reduce intraocular pressure of rabbits, compared with the conventional dose group, pilocarpine+timolol group and timolol pilocarpine+group alkali group of intraocular pressure, intraocular pressure and good control effect,the difference was statistically significant (P<0.05).There was no significant difference between in the adverse reactions between pilocarpine group, timolol group, routine administration group, pilocarpine+timolol group, timolol group and pilocarpine group.ConclusionThe treatment of glaucoma can be treated with pilocarpine and timolol eye drops, pilocarpine+timolol compared with pilocarpine, timolol simple treatment, conventional administration of several modes of administration, from the continuous reduction of intraocular pressure time Point of view, lowering the intraocular pressure longer, fewer adverse reactions, with high therapeutic safety, worthy of clinical promotion and application.

Purslane protects against the reproductive toxicity of carbamazepine treatment in pilocarpine-induced epilepsy model

Objective:To investigate the protective effect of purslane with carbamazepine treatment.Methods:Male albino rats were modulated by pilocarpine to be epileptic.Both the normal and epileptic rats were treated with carbamazepine,purslane or carbamazepine plus purslane,with separate non-treated control groups for both normal and epileptic rats.Results:The data from the current study showed amelioration in amino acids and electrolytes in the epileptic rats treated with purslane and carbamazepine,with this amelioration occurring without decreasing the fertility hormones (testosterone,dehydroepiandrosterone,luteinizing hormone and follicle stimulating hormone).Purslane treatments also prevented the increase in estradiol.The decreased epileptic hyperexcitability with purslane was evidenced by decreased glial fibrillary acidic protein and lipid peroxidation.Conclusions:Natural products like purslane could be used with the highly repetitive drugs like carbamazepine to reduce or prevent its side-effects.

Neuroprotective effect of lithium after pilocarpine-induced status epilepticus in mice

Status epilepticus is the most common serious neurological condition triggered by abnormal electrical activity, leading to severe and widespread cell loss in the brain. Lithium has been one of the main drugs used for the treatment of bipolar disorder for decades, and its anticonvulsant and neuroprotective properties have been described in several neurological disease models. However, the therapeutic mechanisms underlying lithium's actions remain poorly understood. The muscarinic receptor agonist pilocarpine is used to induce status epilepticus, which is followed by hippocampal damage. The present study was designed to investigate the effects of lithium post-treatment on seizure susceptibility and hippocampal neuropathological changes following pilocarpine-induced status epilepticus. Status epilepticus was induced by administration of pilocarpine hydrochloride (320 mg/kg, i.p.) in C57BL/6 mice at 8 weeks of age. Lithium (80 mg/kg, i.p.) was administered 15 minutes after the pilocarpine injection. After the lithium injection, status epilepticus onset time and mortality were recorded. Lithium significantly delayed the onset time of status epilepticus and reduced mortality compared to the vehicle-treated group. Moreover, lithium effectively blocked pilocarpine-induced neuronal death in the hippocampus as estimated by cresyl violet and Fluoro-Jade B staining. However, lithium did not reduce glial activation following pilocarpine-induced status epilepticus. These results suggest that lithium has a neuroprotective effect and would be useful in the treatment of neurological disorders, in particular status epilepticus.

Structural and Functional Alterations at Pre-Epileptic Stage Are Closely Associated with Epileptogenesis in Pilocarpine-induced Epilepsy Model

Pilocarpine-induced rat epilepsy model is an established animal model that mimics medial temporal lobe epilepsy in humans. The purpose of this study was to investigate neuroimaging abnormalities in various stages of epileptogenesis and to correlate them with seizure severity in pilocarpine-induced rat epilepsy model. Fifty male Sprague-Dawley rats were subject to continuous video and electroencephalographic monitoring after inducing status epilepticus (SE) and seizure severity was estimated by frequency and total durations of class 3 to 5 spontaneous recurrent seizures (SRS) by modified Racine's classification. The 7.0 Tesla magnetic resonance imaging (MRI) with high resolution flurodeoxyglucose positron emission tomography (FDG-PET) was performed at 3 hours, 1, 3, 7 days and 4 weeks after the initial insult. The initial SRS was observed 9.7±1.3 days after the pilocarpine injection. MRI revealed an abnormal T2 signal change with swelling in both hippocampi and amygdala in acute (day 1 after injection) and latent phases (days 3 and 7), in association with PET hypometabolism in these areas. Interestingly, the mean frequency of class 3 to 5 SRS was positively correlated with abnormal T2 signals in hippocampal area at 3 days. SRS duration became longer with more decreased glucose metabolism in both hippocampi and amygdala at 7 days after pilocarpine injection. This study indicates that development and severity of SRS at chronic phase could be closely related with structural and functional changes in hippocampus during the latent period, a pre-epileptic stage.

Impaired executive functions in experimental model of temporal lobe epilepsy / Prejuízo das funções executivas no modelo experimental de epilepsia do lobo temporal

ABSTRACT Objective The present study aimed to investigate cognitive and behavioural changes consistent with attention deficit hyperactivity disorder (ADHD -like behavior in male Wistar rats with temporal lobe epilepsy (TLE). Method Male Wistar rats at 25 day of age were submitted to animal model of TLE by pilocarpine injection (350 mg/kg, ip) and a control group received saline 0.9%. The animals were continuously video monitored up to the end of experiments. The behavioural tests (open field, elevated plus maze and operant conditioning box) started from 60 days postnatal. Results Animals with TLE exhibited elevated locomotor activity, reduced level of anxiety-related behavior, impulsivity and impaired visuospatial working memory. Conclusion Taken as a whole, we concluded that animals with TLE exhibited some cognitive and behavioural changes consistent with ADHD-like behavior.

RESUMO Objetivo O presente estudo teve como objetivo investigar as alterações cognitivas e comportamentais consistentes com o comportament de transtorno de deficit de atenção e hiperatividade (TDAH) -like em ratos Wistar machos com epilepsia do lobo temporal (ELT). Método Ratos Wistar machos com 25 dias de vida foram submetidos a modelo animal de ELT pela injeção de pilocarpina (350 mg / kg, ip) e grupo controle recebeu salina 0,9%. Os animais foram monitorados continuamente por vídeo até ao final dos experimentos. Os testes comportamentais (campo aberto, labirinto em cruz elevado e caixa de condicionamento operante) começaram a partir de 60 dias pós-natal. Resultados Os animais com ELT exibiram aumento da atividade locomotora, redução do comportamento relacionado com a ansiedade, impulsividade e prejuízo da memória de trabalho visuospatial. Conclusão Em conjunto, concluímos que os animais com ELT apresentaram algumas alterações cognitivas e comportamentais consistentes com o comportamento TDAH-like.