In vitro and in silico evaluation of the schistosomicidal activity of eugenol derivatives using biochemical, molecular, and morphological tools

Background Eugenol shows both antibacterial and antiparasitic activities, suggesting that it might be evaluated as an option for the treatment of praziquantel-resistant schistosome. Methods The in vitro activities of three eugenol derivatives (FB1, FB4 and FB9) on adult worms from Schistosoma mansoni were examined by fluorescence and scanning electron microscopy to analyze effects on the excretory system and integument damage, respectively. Biochemical tests with verapamil (a calcium channel antagonist) and ouabain (a Na+/K+-ATPase pump inhibitor) were used to characterize eugenol derivative interactions with calcium channels and the Na+/K+-ATPase, while in silico analysis identified potential Na+/K+-ATPase binding sites. Results The compounds showed effective doses (ED50) of 0.324 mM (FB1), 0.167 mM (FB4), and 0.340 mM (FB9). In addition, FB4 (0.322 mM), which showed the lowest ED50, ED90 and ED100 (p < 0.05), caused the most damage to the excretory system and integument, according to both fluorescence and scanning electron microscopy analysis. The death of adult worms was delayed by ouabain treatment plus FB1 (192 versus 72 hours) and FB9 (192 versus 168 hours), but the response to FB4 was the same in the presence or absence of ouabain. Besides, no changes were noted when all of the eugenol derivatives were combined with verapamil. Moreover, FB1 and FB9 inhibited Na+/K+-ATPase activity according to in silico analysis but FB4 did not show a time-dependent relationship and may act on targets other than the parasite Na+/K+-ATPase. Conclusion Eugenol derivatives, mainly FB4 when compared to FB1 and FB9, seem to act more effectively on the integument of adult S. mansoni worms.(AU)

The photobiomodulation (658, 830 and 904nm) on wound healing in histomorphometric analysis / A fotobiomodulação (658, 830 e 904nm) na cicatrização de feridas na análise histomorfométrica / La fotobiomodulación (658, 830 y 904nm) en la cicatrización de heridas en el análisis histomorfométrico

Abstract Introduction: Photobiomodulation (PBM) assists in the processes of angiogenesis and cellular mitosis after skin lesion, contributing to tissue repair. Objective: to investigate the effects of photobiomodulation (during the proliferative phase) of 658 nm, 830 nm and 904 nm in the repair of skin lesions in an animal model. Method: 658 nm (G658), 830 nm (G830), 904 nm (G904) PBM, and control group (CG) integrated the research. We submitted the animals to an excisional wound and treatment at different wavelengths for 14 days. On the seventh and 14-1485004059th postoperative days, we calculated the area and percentage of lesion contraction. The animals were sacrificed on the 14-1485004056th postoperative day and cutaneous section of the injured region was collected for histomorphometric evaluation of the cellularity, neovascularization, thickness of the epidermis and volume density of collagen fibers colored with H&E and Picross Sirius respectively. For the statistical analysis, we applied the ANOVA test. Results: the G658 presented higher cellularity than GC (p = 0.03). The animals in the G658 group showed a significant increase in the neovascularization in relation to the CG (p = 0.01). Type III collagen significantly increased in G904 compared to G830 (p < 0.0001) and CG (p < 0.0001). The G658 had a significant increase in type III collagen fibers compared to G830 (p < 0.0001) and GC (p < 0.0001). We found no significant difference in the thickness of the epidermis, wound area, and in the percentage wound of contraction between the analyzed groups. Conclusion: PBM was effective to stimulate the tissue repair process, with better results for the 658 nm wavelength.

Resumo Introdução: A Fotobiomodulação (FBM) auxilia nos processos de angiogênese e mitose celular após lesão cutânea, contribuindo para reparo do tecido. Objetivo: investigar os efeitos da fotobiomodulação (durante a fase proliferativa) com comprimento de onda de 658 nm, 830 nm e 904 nm no reparo de lesões cutâneas em modelo animal. Método: FBM 658 nm (G658), 830 nm (G830), 904 nm (G904) e controle (GC) integraram a pesquisa. Os animais foram submetidos a uma ferida excisional e receberam tratamento em diferentes comprimentos de por 14 dias. No 7º e 14º dia pós-operatório, calculou-se a área e a porcentagem de contração da lesão. Os animais foram sacrificados no 14º dia pós-operatório e a secção cutânea da região lesada foi coletada para avaliação histomorfométrica da celularidade, neovascularização, espessura da epiderme e densidade volumétrica das fibras colágenas, corados com H&E e Picross Sirius respectivamente. Para a análise estatística, foi aplicado o teste ANOVA. Resultados: o G658 apresentou maior celularidade que GC (p = 0,03). Os animais do grupo G658 apresentaram aumento significativo da neovascularização em relação ao GC (p = 0,01). Houve aumento significativo do colágeno tipo III no G904 em relação ao G830 (p < 0,0001) e GC (p < 0,0001). O G658 teve um aumento significativo nas fibras colágenas tipo III em comparação ao G830 (p < 0,0001) e GC (p < 0,0001). Nenhuma diferença significativa foi encontrada na espessura da epiderme, área da ferida e na porcentagem de contração da ferida entre os grupos analisados. Conclusão: a PBM foi efetiva para estimular o processo de reparo tecidual, com melhores resultados para o comprimento de onda de 658 nm.

Resumen Introducción: La fotobiomodulación (FBM) auxilia en los procesos de angiogénesis y mitosa celular después de lesión cutánea, contribuyendo para la reparación. Objetivo: investigar los efectos de la fotobiomodulación (durante la fase proliferativa) con longitud de onda de 658 nm, 830 nm y 904 nm en la reparación de lesiones cutáneas en modelo animal. Método: grupos de FBM 658 nm (G658), 830 nm (G830), 904 nm (G904) y control (GC) integraron la investigación. Los animales fueron sometidos a una herida excisional y recibieron tratamiento 14 días. En el 7º y 14º día postoperatorio, se calculó el área y el porcentaje de contracción de la lesión. Los animales fueron sacrificados en el 14º día postoperatorio y la sección cutánea de la región lesada fue recolectada para evaluación histomorfométrica de la celularidad, neovascularización, espesor de la epidermis y densidad volumétrica de las fibras colágenas, colorados con H & E y Picross Sirius respectivamente. Para el análisis estadístico, se aplicó la prueba ANOVA. Resultados: G658 presentó mayor celularidad que GC (p = 0,03). G658 presentaron un aumento significativo de la neovascularización en relación al GC (p = 0,01). Se observó un aumento significativo del colágeno tipo III en el G904 con respecto al G830 (p < 0,0001) y GC (p < 0,0001). El G658 tuvo un aumento significativo en las fibras colágenas tipo III en comparación con el G830 (p < 0,0001) y GC (p < 0,0001). Ninguna diferencia significativa se encontró en el espesor de la epidermis, área de la herida entre los grupos. Conclusión: la PBM fue efectiva para estimular el proceso de reparación del tejido, con mejores resultados para grupo 658 nm.

Bark Extract of Bathysa cuspidata in the Treatment of Liver Injury Induced by Carbon Tetrachloride in Rats

The aim of this study was to investigate the effect of bark extract Bathysa cuspidata (BCE) in the reconstitution of hepatic parenchyma and stroma after dysfunction induced by the CCl4. Liver lesions were induced by intraperitoneal administration of CCl4 (1 mL/kg) every 48 h for 12 days. The animals were treated with B. cuspidata extract (BCE) administered by gavage for another 12 days. Forty-nine rats were randomized into seven treatment groups with seven animals receiving CCl4, BCE (200, or 400 mg/kg) and the vehicle DMSO alone, or in different combinations. The extract alone showed no evidence of hepatic toxicity. In general, rats acutely exposed to CCl4 without the treatment with BCE presented high ALT and AST serum levels and high tissue content of lipid hydroperoxides, malondialdehyde, and lipid droplets. BCE administration, especially at 400 mg/kg attenuated significantly all these parameters. Furthermore, the antioxidant activities of the enzymes superoxide dismutase and catalase were significantly increased in the groups receiving this extract. These results showed that the extract of B. cuspidata stem bark stimulated the antioxidant defense system and reduced the morphological and functional liver damage in Wistar rats previously exposed to CCl4.

Exercise training prior to myocardial infarction attenuates cardiac deterioration and cardiomyocyte dysfunction in rats

OBJECTIVES: The present study was performed to investigate 1) whether aerobic exercise training prior to myocardial infarction would prevent cardiac dysfunction and structural deterioration and 2) whether the potential cardiac benefits of aerobic exercise training would be associated with preserved morphological and contractile properties of cardiomyocytes in post-infarct remodeled myocardium. METHODS: Male Wistar rats underwent an aerobic exercise training protocol for eight weeks. The rats were then assigned to sham surgery (SHAM), sedentary lifestyle and myocardial infarction or exercise training and myocardial infarction groups and were evaluated 15 days after the surgery. Left ventricular tissue was analyzed histologically, and the contractile function of isolated myocytes was measured. Student's t-test was used to analyze infarct size and ventricular wall thickness, and the other parameters were analyzed by the Kruskal-Wallis test followed by Dunn's test or a one-way analysis of variance followed by Tukey's test (p<0.05). RESULTS: Myocardial infarctions in exercise-trained animals resulted in a smaller myocardial infarction extension, a thicker infarcted wall and less collagen accumulation as compared to myocardial infarctions in sedentary animals. Myocardial infarction-induced left ventricular dilation and cardiac dysfunction, as evaluated by +dP/dt and -dP/dt, were both prevented by previous aerobic exercise training. Moreover, aerobic exercise training preserved cardiac myocyte shortening, improved the maximum shortening and relengthening velocities in infarcted hearts and enhanced responsiveness to calcium. CONCLUSION: Previous aerobic exercise training attenuated the cardiac dysfunction and structural deterioration promoted by myocardial infarction, and such benefits were associated with preserved cardiomyocyte morphological and contractile properties. .

Uso de fluorescência em um método de dissector modificado para estimar o número de miócitos no tecido cardíaco / Use of fluorescence in a modified disector method to estimate the number of myocytes in cardiac tissue / Uso de fluorescencia en un método de disector modificado para estimar el número de miocitos en el tejido cardíaco

FUNDAMENTO: Métodos convencionais de dissector atualmente requerem consideráveis custos financeiros, técnicos e operacionais para estimar o número de células, incluindo cardiomiócitos, em uma área de 3D. OBJETIVO: Usar a microscopia de fluorescência em um método de dissector modificado para determinar o número de miócitos no tecido cardíaco em condições normais e patológicas. MÉTODOS: O estudo empregou camundongos Wistar machos com quatro meses de idade e peso de 366,25 ± 88,21 g randomizados em grupos controles (GC, n = 8) e infectados (GI, n = 8). Os animais do GI foram inoculados com cepa Y de T. cruzi (300.000 tripomastigotas/50 g). Após oito semanas, os animais foram pesados e sacrificados. Os Ventrículos Esquerdos (VE) foram removidos para análise estereológica da densidade numérica de cardiomiócitos (Nv [c]) e o número total dessas células no VE (N [c]). Esses parâmetros foram estimados usando um dissector fluorescente (DF) e comparados com os métodos convencionais de dissector óptico (DO) e dissector físico (DFi). RESULTADOS: Em ambos os métodos de dissector, os animais do GI apresentaram queda significativa de Nv[c] e N[c] em comparação com os animais do GC (P > 0,05). Uma correlação forte, igual ou superior a 96%, foi obtida entre DF, DO e DFi. CONCLUSÃO: O método DF parece ser igualmente confiável para determinar Nv[c] e N[c] em condições normais e patológicas, apresentando algumas vantagens em relação aos métodos convencionais de dissector: redução de cortes histológicos e imagens na análise estereológica, redução do tempo de análise das imagens, a construção de DF em microscópios simples, utilizando o modo de epifluorescência, distinção de planos de dissector em ampliações inferiores.

BACKGROUND: Conventional disector methods currently require considerable financial, technical and operational costs to estimate the number of cells, including cardyomyocytes, in a 3D area. OBJECTIVE: To use fluorescence microscopy in a modified disector method to determine the number of myocytes in cardiac tissue in normal and pathological conditions. METHODS: The study employed four-month-old male Wistar rats with weight of 366.25 ± 88.21g randomized in control (CG, n=8) and infected (IG, n=8) groups. IG animals were inoculated with T. cruzi Y strain (300,000 trypomastigotes/50g wt). After eight weeks, the animals were weighted and euthanized. The left ventricles (LV) were removed for stereological analysis of numerical density of cardiomyocytes (Nv[c]) and total number of these cells in the LV (N[c]). These parameters were estimated using a fluorescent disector (FD) and compared with the conventional optical (OD) and physical (PD) disector methods. RESULTS: In both disector methods, IG animals presented significant decrease of Nv[c] and N[c] compared to CG animals (P< 0.05). There was no significant difference in these variables despite the disector method applied in CG and IG animals (P> 0.05). A strong correlation, equal or above 96%, was obtained between FD, OD and PD. CONCLUSION: The FD method seems to be equally reliable to determine Nv[c] and N[c] in normal and pathological conditions and presents some advantages compared to conventional disector methods: reduction of histological slices and images in the stereological analysis, reduction of time to analyze the images, construction of FD in simple microscopes using the epifluorescence mode, distinction of disector planes in lower magnifications.

FUNDAMENTO: Métodos convencionales de disector actualmente requieren considerables costos financieros, técnicos y operativos para estimar el número de células, incluyendo cardiomiocitos, en un área de 3D. OBJETIVO: Usar la microscopia de fluorescencia en un método de disector modificado para determinar el número de miocitos en el tejido cardíaco en condiciones normales y patológicas. MÉTODOS: El estudio empleó ratones Wistar machos de cuatro meses de edad y peso de 366,25 ± 88,21 g randomizados en grupos controles (GC, n = 8) e infectados (GI, n = 8). Los animales del GI fueron inoculados con cepa Y de T. cruzi (300.000 tripomastigotas/50 g). Después de ocho semanas, los animales fueron pesados y sacrificados. Los Ventrículos Izquierdos (VI) fueron removidos para análisis estereológico de la densidad numérica de cardiomiocitos (Nv [c]) y el número total de esas células en el VI (N [c]). Esos parámetros fueron estimados usando un disector fluorescente (FD) y comparados con los métodos convencionales de disector óptico (OD) y disector físico (PD). RESULTADOS: En ambos métodos de disector, los animales del GI presentaron caída significativa de Nv[c] y N[c] en comparación con los animales del GC (P > 0,05). Una correlación fuerte, igual o superior a 96%, fue obtenida entre FD, OD y PD. CONCLUSIÓN: El método FD parece ser igualmente confiable para determinar Nv[c] y N[c] en condiciones normales y patológicas, presentando algunas ventajas en relación a los métodos convencionales de disector: reducción de cortes histológicos e imágenes en el análisis estereológico, reducción del tiempo de análisis de las imágenes, la construcción de FD en microscopios simples, utilizando el modo de epifluorescencia, distinción de planos de disector en ampliaciones inferiores.

Equações de referência para a predição da força de preensão manual em brasileiros de meia idade e idosos / Reference equations or predicting of handgrip strength in Brazilian middle-age and elderly subjects

O objetivo deste estudo foi avaliar os valores normais da força de preensão manual do membro superior dominante (FPM-D) e não dominante (FPM-ND) em sujeitos de meia idade e idosos assintomáticos e elaborar equações de referência para a predição da FPM. Foram investigados 54 voluntários (51,9% homens) com idade ≥50 anos, medindo-se massa corporal, estatura e perimetria do braço direito e esquerdo, e calculando-se o índice de massa corporal. A FPM-D e FPM -ND foram avaliadas por dinamometria mecânica. O índice de atividade física habitual (IAF) foi avaliado pelo questionário de Baecke. A FPM-D foi superior à FPM-ND em ambos os sexos e em todas as idades (p<0,05). Foram encontradas correlações significativas entre a FPM e idade, estatura, massa corporal e perimetria do braço. As melhores equações de referência foram as seguintes: FPM-Dkgf =39,996 - (0,382 x idade anos)+(0,174 x peso kg)+(13,628 x sexo homens=1; mulheres=0) (R2 ajustado=0,677); eFPM-NDkgf=44,968- (0,420 x idade anos)+(0,110 x peso kg)+(9,274 x sexo homens=1; mulheres=0) (R2 ajustado=0,546). A diferença consistente entre a FPM-D e FPM-ND torna necessário o uso de dados normativos específicos para cada mão. Atributos simples de serem obtidos, tais como idade, estatura, massa corporal, perimetria do braço e sexo, podem pois prever adequadamente os valores esperados da FPM para adultos e idosos assintomáticos...

The aim of the present study was to evaluate normal values of dominant (D) and non-dominant (ND) upper limb handgrip strength (HGS) in asymptomatic middle-aged elderly subjects and to establish reference equations for predicting HGS. Fifty-four volunteers (51.9% men) aged ≥50 years old were enrolled. Weight, height, and both arms circumference were measured, being the body mass index calculated. Mechanical dynamometry was used to measure D-HGS and ND-HGS. Self-reported level of regular physical activity was assessed by Baecke questionnaire. D-HGS was higher than ND-HGS for both sexes and in all age groups (p<0.05). Significant correlations were found between HGS and age, height, weight and arm circumferences. The best reference equations were the following: D-HGSkgf=39.996 - (0.382 x age years)+(0.174 x weight kg)+(13.628 x sex men=1; women=0) (Adjusted R2=0.677); and ND-HGS kgf=44.968 - (0.420 x age years)+(0.110 x weight kg)+(9.274 x sex men=1; women=0) (Adjusted R2=0.546). The consistent difference found between dominant and non-dominant HGS requires the use of specific normative data for each hand. Hence easily-obtained attributes such as age, height, weight, arm circumference and sex can predict HGS expected values for asymptomatic elder adults...

Ombro doloroso após lesão cerebral / Painful shoulder after brain injury

O ombro doloroso (OD) após acidente vascular encefálico (AVE) é frequentemente investigado. Entretanto, sua prevalência após outros tipos de lesão cerebral permanece desconhecida. Os objetivos deste estudo foram identificar a prevalência de OD após diferentes tipos de lesões cerebrais, os fatores associados a essa condição em pacientes admitidos na reabilitação e investigar a associação entre suas características clínicas. Foram avaliados os prontuários do universo de pacientes (n = 109) com diagnóstico clínico e imaginológico de LC admitidos no período de um ano para tratamento em um hospital da rede pública de saúde de Minas Gerais. Itens como, idade, gênero, etiologia da lesão, tempo de lesão, deficiência física, ombro acometido, tônus muscular, subluxação glenoumeral, amplitude de movimento (ADM) e distrofia simpático-reflexa (DSR) foram incluídos neste estudo. A prevalência de ombro doloroso foi de 52,29 por cento. Dentre os fatores previamente relacionados, apenas o tempo de lesão em meses, espasticidade grau 1 na escala de Ashworth, limitação da ADM e subluxação glenoumeral apresentaram associação significativa com essa complicação. As manifestações clínicas identificadas nos prontuários com registro de OD apresentaram significância apenas para a associação do tipo de tônus muscular com o grau de tônus e a ADM.

The painful shoulder after stroke is often investigated. However, its prevalence after other types of brain injury remains unknown. The objectives of this study were to identify the prevalence of painful shoulder after different types of brain injury, the factors associated with this condition in patients admitted to the rehabilitation and investigate the association between its clinical features. The records were evaluated of the universe of patients (n = 109) with clinical and imaginologic diagnosis of brain injury admitted within a year for treatment at a general hospital of Minas Gerais. Items such as age, gender, etiology and time of injury, disability, affected shoulder, muscle tone, glenohumeral subluxation, range of motion and reflex sympathetic dystrophy were included in this study. The prevalence of shoulder pain was 52.29 percent. Among the factors previously listed, only the time of injury in months, spasticity grade 1 on the scale of Ashworth, limitation of range of motion and glenohumeral subluxation showed significant association with this complication. Clinical manifestations identified in records with shoulder painful had significance only for the combination of the type of tone to the degree of muscle tone and range of motion.