Association of Rhizophora mangle and ascorbic acid in hydrogels: Evaluation of cytotoxic and immunomodulatory effects

Abstract Polyphenolics from Rhizophora mangle (R. mangle) have shown wound healing and anti- inflammatory effects that may be potentiated by being associated with ascorbic acid, an important substance for collagen and elastin synthesis that plays a role in tissue repair. In our study, we aimed to formulate an association of R. mangle and ascorbic acid in hydrogels and evaluate the association's cytotoxic and immunomodulatory effects. In a pre-formulation study, three gelling polymers (i.e.xanthan gum, poloxamer and hydroxyethyl cellulose) were tested. The selected polymer (i.e. xanthan gum) was used to evaluate cytotoxic and immunomodulatory effects using flow cytometry. Xanthan gum (1.5%) had a homogeneous appearance, an orange colour, a smooth surface, intense brightness and the typical odour, as well as non-Newtonian pseudoplastic behaviour. With a pH of 5.0-5.3 and a non-cytotoxic profile, xanthan gum induced the proliferation and activation of CD4 +, CD8+ and NK T lymphocytes and the production of IL- 2, IL-4, IL-10, IL-17 and TNF-α cytokines in stimulated splenocytes. The results suggest that the association of R. mangle and ascorbic acid in 1.5% xanthan gum hydrogel may be promising in preparations for wound-healing processes.

Pectin-like Polysaccharide Extracted from the Leaves Caesalpinia pulcherrima is a Promising Antioxidant and Immunomodulator Agent

Abstract Studies have shown that Caesalpinia pulcherrima extracts promote antioxidant, healing, immunomodulating and antiparasitic activities and its polysaccharides can be used as functional food. In this sense, this work had as objective the isolation and characterization of a polysaccharide-like pectin, extracted from the C. pulcherrima leaves and its possible applications as an antioxidant and immunomodulator agent. The molecule was characterized by high performance liquid chromatography, fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Its antioxidant potential was evaluated through the methods of phosphomolybdenum, ABTS radical scavenging [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid], DPPH (1,1-diphenyl-2-picrylhydrazyl) and nitric oxide radical. The immunostimulating effects of pectin were tested in splenocytes to evaluate its toxic, proliferative and cell activator and immunomodulatory potential. The polysaccharide obtained has structural characteristics similar to pectins. Pectin showed high in vitro antioxidant activity for ABTS radical scavenging, moderate activity for phosphomolybdenum and low activity for DPPH and nitric oxide. In vitro immunomodulation assays showed that pectin obtained did not promote a cytotoxic effect (viability > 90%). The increase in cytosolic ROS levels indicates a possible mechanism of cell activation without causing damage. Immunophenotyping showed that pectin increased a subpopulation of CD8+ T lymphocytes and monocytes. In addition, it promoted a mostly pro-inflammatory response confirmed by the production of cytokines IL-2, -4, -6, IFN-γ and TNF-α. These results reinforce the ethnopharmacological use of C. pulcherrima leaves and expand the use of this plant for future applications as herbal medicines.

A proposed eye ex vivo permeation approach to evaluate pesticides: Case dimethoate.

This study aims to propose different ex vivo methodologies for pesticide evaluation when in contact with ocular mucosa. The first ex vivo study was performed using vertical Franz cells with fresh and refrigerated excised bovine corneas. The second evaluated the permeation through the cornea by using fresh, refrigerated and damaged whole bovine eyes. Both experiments were evaluated by applying 50 µl (of 5 mg/ml solution) of an emulsifiable pesticide formulation (Dimetoato 500 EC®). In the first study, dimethoate profiles and permeation fluxes showed no statistical differences (p < .05) between fresh and refrigerated excised corneas, probably due to the excision procedure generating damage to the cornea by altering stromal structure, irrespective of the refrigeration procedure. In relation to the results obtained for developed ex vivo permeation method in whole eyes, there was a statistical difference (p < .05) between experiments performed with fresh eyes and those performed with refrigerated/damaged eyes. Damage generated by both the refrigeration process and exposure to irritant products led to a five-fold reduction in the amount of corneal-permeated pesticide as well as a two-fold increase in retention. Thus, the ex vivo permeation approach with whole eyes seems to be a simple and reproducible method to evaluate pesticides. It is evident that further evaluations need to be performed using other pesticides with distinct physicochemical characteristics.

Synthesis, in vitro and in vivo biological evaluation, COX-1/2 inhibition and molecular docking study of indole-N-acylhydrazone derivatives.

The objective of this work was to obtain and evaluate anti-inflammatory in vitro, in vivo and in silico potential of novel indole-N-acylhydrazone derivatives. In total, 10 new compounds (3a-j) were synthesized in satisfactory yields, through a condensation reaction in a single synthesis step. In the lymphoproliferation assay, using mice splenocytes, 3a and 3b showed inhibition of lymphocyte proliferation of 62.7% (±3.5) and 50.7% (±2), respectively, while dexamethasone presented an inhibition of 74.6% (±2.4). Moreover, compound 3b induced higher Th2 cytokines production in mice splenocytes cultures. The results for COX inhibition assays showed that compound 3b is a selective COX-2 inhibitor, but with less potency when compared to celecoxib, and compound 3a not presented selectivity towards COX-2. The molecular docking results suggest compounds 3a and 3b interact with the active site of COX-2 in similar conformations, but not with the active site of COX-1, and this may be the main reason to the COX-2 selectivity of compound 3b. In vivo carrageenan-induced paw edema assays were adopted for the confirmation of the anti-inflammatory activity. Compound 3b showed better results in suppressing edema at all tested concentrations and was able to induce an edema inhibition of 100% after 5 h of carrageenan injection at the 30 mg kg-1 dosage, corroborating with the COX inhibition and lymphoproliferation results. I addition to our experimental results, in silico analysis suggest that compounds 3a and 3b present a well-balanced profile between pharmacodynamics and pharmacokinetics. Thus, our preliminary results revealed the potentiality of a new COX-2 selective derivative in the modulation of the inflammatory process.

Use of complex decongestive therapy with low cost material in a patient with lymphedema living in an endemic area for filariasis

Lymphatic filariasis affects millions of people around the world and may have disabling consequences. Lymphedema stands out as a chronic manifestation in patients with this disease, affecting the lower limbs and limiting motor function. This study aimed to propose a treatment protocol to decrease lower limb lymphedema of treated patients and to show the efficacy of Complex Decongestive Therapy (CDT) as an auxiliary resource for this condition. This is a case study of a patient treated with CDT, making use of compression methods with alternative materials and instructions for asepsis during 10 weeks. The results demonstrated that this treatment reduced lymphedema in the right lower limb of the patient from grade V to grade III, with areas of measurement reduction of up to 41%. In addition, acute dermatolymphangioadenitis (ADLA) manifestations stopped in the treated patient. CDT associated with low-cost compression material, has been shown to be effective in reducing lower limb lymphedema and ADLA crises, as well as in promoting improved locomotion and performance of daily activities by the patient

Perfil de pacientes com linfedema atendidos no serviço de referência nacional em filarioses da Fundação Oswaldo Cruz, Pernambuco, Brasil / Profile of lymphedema patients attended at the national center of lymphatic filariasis of the Oswaldo Cruz Foundation, Pernambuco, Brazil

A filariose linfática é uma parasitose endêmica em 73 países e afeta, aproximadamente, 120 milhões de pessoas em todo o mundo. Este trabalho buscou traçar o perfil de pacientes com linfedema que residem em áreas endêmicas de filariose, com ou sem diagnóstico comprovado de doença prévia. Trata-se de um estudo transversal que envolveu pacientes com linfedema, cadastrados no SRNF-CPqAM-FIOCRUZ-PE, residentes nas principais cidades da região metropolitana de Recife, capital do estado de Pernambuco, aos quais foi aplicado um questionário para avaliação do perfil sociodemográfico e epidemiológico. Foram selecionados 35 pacientes, sendo 80% do sexo feminino, a maioria com idade entre 40 e 59 anos (48,6%). Do total examinado, 40% apresentaram linfedema bilateral de membros inferiores e presença de dor moderada no membro afetado. Além disso, dos indivíduos atendidos, apenas 20% receberam diagnóstico de filariose e 68% relataram morar em ruas pavimentadas com disponibilidade de coleta regular de lixo e acesso à água potável. Os resultados deste estudo propiciaram um conhecimento mais abrangente do perfil dos pacientes com linfedema atendidos pelo serviço de referência e poderão auxiliar no aprimoramento das políticas públicas de saúde.

Terapia complexa descongestiva com uso de material alternativo na redução e controle do linfedema em pacientes de área endêmica de filariose: um ensaio clínico / Terapia compleja descongestiva con el empleo de material alternativo reduce y controla el linfedema en pacientes de zonas endémicas de filariasis: estudio clínico / Complex decongestant therapy with use of alternative material to reduce and control lymphedema in patients with endemic area of filariasis: a clinical trial

RESUMO A filariose linfática põe em risco bilhões de pessoas em todo o mundo. Apesar disso, pouco se sabe sobre o impacto da morbidade crônica da doença, como o linfedema. A terapia complexa descongestiva apresenta-se como padrão ouro para tratar linfedemas, mas ainda não existem protocolos viáveis para implementação em países em desenvolvimento. O objetivo deste ensaio clínico foi avaliar a eficácia dessa técnica, com o uso de material alternativo, e comparar seus efeitos na qualidade de vida, funcionalidade e peso corporal. Os grupos intervenção e controle foram avaliados por meio de perimetria e cálculo do volume dos membros, do questionário de qualidade de vida Whoqol-bref, do teste de funcionalidade e mobilidade de membros inferiores Timed Up and Go e avaliação do peso corporal. O grupo intervenção recebeu a terapia complexa descongestiva duas vezes por semana, durante dez semanas, utilizando-se um material alternativo para o enfaixamento, confeccionado com tecido de cambraia, o que apresenta baixo custo em relação às faixas importadas. O grupo controle recebeu palestra com informações sobre a doença e orientações de cuidados e higiene dos membros. A amostra foi composta por trinta pacientes com idade média de 50,8±10 anos, sendo 52,9% mulheres. Na análise estatística foram utilizados os testes t de Student, Multivariate analysis of variance, teste de Wilcoxon e Kolmogorov-Smirnov. A significância adotada foi de 5% (p<0,05). Houve redução significativa do volume e perimetria do linfedema no grupo intervenção, e aumentou no grupo controle. A funcionalidade não apresentou melhora significativa na avaliação pelo teste Timed Up and Go. A qualidade de vida teve melhora significativa nos domínios físico e meio ambiente no grupo intervenção. O efeito do tratamento no peso corporal também foi significativo, apresentando redução no grupo controle e intergrupos. A terapia complexa descongestiva mostrou-se eficaz na redução e no controle do linfedema e impactou de maneira positiva, aumentando os valores numéricos dos aspectos físico e meio ambiente da qualidade de vida do grupo intervenção.

RESUMEN La filariasis linfática presenta riesgo para muchas personas en el mundo. Pero poco se sabe acerca del impacto de la morbilidad crónica de esta enfermedad, como es el linfedema. Y surge la terapia compleja descongestiva como la forma de tratarlo, pero todavía no hay protocolos que le posibilite su implementación en países en desarrollo. Este estudio clínico tiene el propósito de valorar la eficacia de esta técnica, con el empleo de material alternativo, y de comparar sus efectos en la calidad de vida, funcionalidad y peso corporal. Se evaluaron a los grupos intervención y control a través de medición y cálculo del volumen de los miembros, de cuestionario de calidad de vida Whoqol-Bref, de la prueba de funcionalidad y movilidad de los miembros inferiores Timed Up and Go y la evaluación del peso corporal. El grupo intervención hizo la terapia compleja descongestiva dos veces a la semana, durante diez semanas, y utilizó un material alternativo, el linón, para el vendaje, lo que mostró ser de bajo costo en comparación a otras vendas importadas. El grupo control vio una charla que exponía informaciones sobre la enfermedad y sobre los cuidados e higienización de los miembros. Treinta pacientes con promedio de edad de 50,8±10 años formaron la muestra, con un 52,9% de participantes mujeres. En el análisis estadístico se emplearon las pruebas t de Student, Multivariate analysis of variance, la prueba de Wilcoxon y Kolmogorov-Smirnov. El nivel de significación ha sido de 5% (p<0,05). Se observó una significativa reducción en el volumen y en la medición del linfedema en el grupo intervención, pero había aumentado en el grupo control. La funcionalidad no ha presentado mejora en la evaluación por la prueba Timed Up and Go. La calidad de vida ha presentado mejora en los dominios físico y medioambiente en el grupo intervención. El efecto del tratamiento en el peso corporal también ha sido significativo por presentar reducción en el grupo control e intergrupal. La terapia compleja descongestiva puede ser eficaz en la reducción y en el control del linfedema, además de aumentar positivamente los valores en los dominios físico y medioambiente de la calidad de vida del grupo intervención.

ABSTRACT Lymphatic filariasis puts billions of people around the world at risk. Despite this, little is known about the impact of chronic disease morbidity, such as lymphedema. The complex decongestant therapy is a gold standard for treating lymphedemas, but there are no viable protocols for implementation in developing countries. The objective of this clinical trial was to evaluate the efficacy of this technique with the use of alternative material and compare its effects on quality of life, functionality and body weight. The intervention and control groups were evaluated using perimetry and limb volume calculation, the Whoqol-bref quality of life questionnaire, the Functional and Mobility test of the lower limbs Timed Up and Go, and body weight assessment. The intervention group received the complex decongestant therapy twice a week for ten weeks, using an alternative material for the bandaging, made with cambric tissue, which presents low cost in relation to the imported bands. The control group received a lecture with information about the disease and care and hygiene guidelines of the members. The sample consisted of thirty patients with a mean age of 50.8 ± 10 years, 52.9% of whom were women. In the statistical analysis, Student's t test, Multivariate analysis of variance, Wilcoxon test and Kolmogorov-Smirnov test were used. The significance was 5% (p<0.05). There was a significant reduction in lymphedema volume and perimetry in the intervention group, and increased in the control group. The functionality did not show significant improvement in the evaluation by the Timed Up and Go test. The quality of life had significant improvement in the physical and environmental domains in the intervention group. The effect of treatment on body weight was also significant, presenting reduction in the control group and intergroups. Complex decongestant therapy was effective in reducing and controlling lymphedema and positively impacted, increasing the numerical values of the physical and environmental aspects of the quality of life of the intervention group.

Immunomodulatory response of mice splenocytes induced by RcaL, a lectin isolated from cobia fish (Rachycentron canadum) serum.

This work reports the isolation of a serum lectin from cobia fish (Rachycentron canadum) named RcaL. Immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production were also performed. RcaL was obtained through precipitation with ammonium sulphate and affinity chromatography on a Concanavalin A-Sepharose 4B column. The ammonium sulphate fraction F3 showed the highest specific hemagglutinating activity and was applied to affinity chromatography. The lectin was eluted with methyl-α-D-mannopyranoside. RcaL showed highest affinity for methyl-α-D-mannopyranoside and D-mannose; eluted fractions of RcaL agglutinated rabbit erythrocytes (titre, 128(-1)) retained 66 % of chromatographed lectin activity, and the obtained purification factor was 1.14. Under reducing conditions, a polypeptide band of 19.2 kDa was revealed in sodium dodecyl sulphate polyacrylamide gel electrophoresis (PAGE). PAGE confirmed RcaL as an acidic protein revealed in a single band. Cytotoxic and immunomodulatory assays with RcaL in mice splenocyte cultures showed that the lectin was not cytotoxic and induced higher interferon gamma and nitric oxide production in splenocyte cultures. Purified RcaL induced preferential Th1 response, suggesting that it acts as an immunomodulatory compound.

Potential effects of Cramoll 1,4 lectin on murine Schistosomiasis mansoni.

Cratylia mollis is a natural forage plant from the Northeast of Brazil. C. mollis seed lectin (Cramoll) containing molecular forms 1 and 4 (Cramoll 1,4) has shown anti-inflammatory and wound-healing activities. This work analyzed the effect of Cramoll 1,4 on experimental schistosomiasis in mice. Experimental groups (n=15/group) were composed of female albino Swiss mice, which were subcutaneously and caudally infected with Schistosoma mansoni (BH strain, 100 cercariae/mouse) and were treated with an intraperitoneal dose after infection as follows: (1) Cramoll 1,4 (50 mg kg(-1) single dose - after 40 days of infection), (2) Cramoll 1,4 (7 mg kg(-1) daily dose - for 7 days after infection) and control (untreated mice). Mice were sacrificed 8 weeks after infection and adult worms were recovered from the portal-hepatic system. Livers were fixed in 10% (v/v) formaldehyde/0.15M NaCl and tissue sections were processed for haematoxilin and Masson's trichrome stainings. Mice infected subcutaneously harboured no or very few worms and hence the effect of Cramoll 1,4 could not be assessed. Results (P≤0.05) were obtained with Cramoll 1,4 using the two treatments, with reduction of: egg excretion (79 and 80%), adult worm recovery (71 and 79%) and liver granulomas (40 and 73.5%) in relation to control. This study showed the potential anti-helminthic activity of Cramoll 1,4 when tested against Schistosomiasis mansoni infection in mice.