Effectiveness of different methods for the extraction of principle actives and phytochemicals content in medicinal herbals / Efectividad de diferentes métodos para la extracción de principios activos y contenido de fitoquímicos en hierbas medicinales

In this present study, we investigated the influence of various extraction methods including maceration, sonication, infusion, decoction, and microwave extraction, on the chemical and biological potential of phytochemicals extracted from three medicinal plants (Ageratum conyzoides, Plantago majorand Arctium lappa L). The results were subsequently analyzed by variance analysis. Our results suggested that sonication is the most effective extraction method among the five methods tested herein, for the extraction of phytochemicals that have a high antioxidant potential and high phenolic content. The three plants employed for this study had a high concentration of flavonoids and phenolics which was compatible with the chemosystematics of the species. All the samples possessed a Sun Protection Factor (SPF) of less than 6. Interestingly, a maximum reaction time of approximately 20 min was noted for the complexation of AlCl3 with the flavonoids present in the phytochemical extract during analyses of the kinetic parameters. We finally identified that the Ageratum conyzoides extract, prepared by sonication, possessed a significant pharmacological potential against hepatocarcinoma tumour cells, whose result can guide further studies for its therapeutic efficacy.

En el presente estudio, investigamos la influencia de varios métodos de extracción, incluyendo maceración, sonicación, infusión, decocción y extracción por microondas, sobre el potencial químico y biológico de los fitoquímicos extraídos de tres plantas medicinales (Ageratum conyzoides, Plantago majory Arctium lappa L). Los resultados se analizaron posteriormente mediante análisis de varianza. Nuestros resultados sugieren que la sonicación es el método de extracción más eficaz entre los cinco métodos aquí probados, para la extracción de fitoquímicos que tienen un alto potencial antioxidante y un alto contenido fenólico. Las tres plantas empleadas para este estudio tenían una alta concentración de flavonoides y fenólicos que era compatible con la quimiosistemática de las especies. Todas las muestras poseían un factor de protección solar (SPF) menor a 6. Curiosamente, se observó un tiempo máximo de reacción de aproximadamente 20 min para la complejación de AlCl3con los flavonoides presentes en el extracto fitoquímico durante los análisis de los parámetros cinéticos. Finalmente, identificamos que el extracto de Ageratum conyzoides, elaborado por sonicación, posee un importante potencial farmacológico frente a las células tumorales del hepatocarcinoma, cuyo resultado puede orientar nuevos estudios sobre su eficacia terapéutica.

Phagolysosomal activity of macrophages in Nile tilapia (Oreochromis niloticus) infected in vitro by Aeromonas hydrophila: Infection and immunotherapy.

The biochemical mechanisms involved in phagocytosis and the intracellular survival of Aeromonas hydrophila (Ah) in host macrophages (MΦs) are complex processes that affect infection success or failure. Thus, in the present study, we described the in vitro infection of Nile tilapia MΦs by a homologous bacterium and tested the effects of anti-A. hydrophila immunoglobulin Y (IgY) on the phagolysosomal activity and intracellular survival of the pathogen. The anti-Ah IgY modulated lysosomal acid phosphatase (LAP) activity as well as the production of reactive oxygen intermediates (ROIs) and nitric oxide (NO), thereby potentiating phagocytosis and the elimination of Ah. Thus, we assume that the specific IgY had a beneficial effect on infection control and postulated the use of the Nile tilapia MΦs as an important in vitro experimental model for the functional and therapeutic study of Ah infection.

Analysis of Fluorescence Spectra of Citrus Polymethoxylated Flavones and Their Incorporation into Mammalian Cells.

Citrus polymethoxylated flavones (PMFs) influence biochemical cascades in human diseases, yet little is known about how these compounds interact with cells and how these associations influence the actions of these compounds. An innate attribute of PMFs is their ultraviolet-light-induced fluorescence, and the fluorescence spectra of 14 PMFs and 7 PMF metabolites were measured in methanol. These spectra were shown to be strongly influenced by the compounds' hydroxy and methoxy substituents. For a subset of these compounds, the fluorescence spectra were measured when bound to human carcinoma Huh7.5 cells. Emission-wavelength maxima of PMF metabolites with free hydroxyl substituents exhibited 70-80 nm red shifts when bound to the Huh7.5 cells. Notable solvent effects of water were observed for nearly all these compounds, and these influences likely reflect the effects of localized microenvironments on the resonance structures of these compounds when bound to human cells.

Electrochemical immunosensor based on ZnO nanorods-Au nanoparticles nanohybrids for ovarian cancer antigen CA-125 detection.

In this research, ZnO nanorods - Au nanoparticles nanohybrids have been fabricated and employed to sensitive electrochemical strategy for the specific detection of the ovarian cancer antigen CA-125/MUC126. The microdevice was developed in our lab based on gold and silver electrodes sputtered on glass substrate. The ZnO nanorods arrays were grown on working electrode using assisted microwave hydrothermal synthesis than gold nanoparticles (Au NPs) were deposited by sputtering. The Au NPs onto ZnO nanorods surface provides a favorable platform for efficient loading of anti-CA-125 antibody via binding with cystamine and glutaraldehyde. The effective loading of the biological material (CA-125 antibody and antigen) on the matrix was observed by SEM images. The electrochemical immunosensor shows a sensitive response to ovarian cancer antigen recombinant human CA-125/MUC126 with detection of 2.5ng/µL, 100 times lower than immunoblot system. Due to high specificity, reproducibility and noteworthy stability, the developed sensor will provide a sensitive, selective and convenient approach to be used to detect CA-125/MUC126.

Improvement of antischistosomal activity of praziquantel by incorporation into phosphatidylcholine-containing liposomes.

Praziquantel (PZQ) is effective against all known species of Schistosomes that infect humans. The failure of mass treatment of schistosomiasis has been attributed to the fact that therapy is not sufficiently long-lasting. This effect may be due to the low bioavailability of PZQ that has a low hydrosolubility and fast metabolism. Liposomes have been used to prolong drug levels, reduce the side effects, direct drugs to specific sites and increase bioavailability after administration. The aim of this work was to study the effect of phosphatidylcholine (PC)-containing liposomes to vehiculate PZQ to improve the treatment of schistosomiasis. The in vitro study was carried out using Schistosoma mansoni parasites recovered by perfusion from the hepatic portal system of infected mice. Suspensions of liposomes with PZQ and free PZQ were administered p.o. in mice after 14 days of infection. The effect of both preparations in vitro on S. mansoni culture was similar. In the in vivo test, PZQ-liposomes caused a decrease in amounts of eggs and parasites. Liposomes improve the antischistosomal activity of praziquantel. This can be used as a starting point to investigate alternative administration routes or dosage forms and to examine the mechanism of intestinal absorption of PRZ.

Abnormalities in apolipoprotein and lipid levels in an HIV-infected Brazilian population under different treatment profiles: the relevance of apolipoprotein E genotypes and immunological status.

HIV infection is associated with disturbances in lipid metabolism due to a host's response mechanism and the current antiretroviral therapy. The pathological appearance and progression of atherosclerosis is dependent on the presence of injurious agents in the vascular endothelium and variations in different subsets of candidate genes. Therefore, the Hha I polymorphism in the apolipoprotein E gene was evaluated in addition to triglycerides, total cholesterol, very low-density lipoprotein (VLDL), LDL, high-density lipoprotein (HDL), and apolipoprotein (apo) Al, B and E levels in 86 Brazilian HIV-infected patients and 29 healthy controls. The allele frequency for apoE in the HIV-infected group and controls was in agreement with data on the Brazilian population. Dyslipidemia was observed in the HIV group and verified by increased levels of triglycerides, VLDL and apoE, and decreased levels of HDL and apoAl. The greatest abnormalities in these biochemical variables were shown in the HIV-infected individuals whose immune function was more compromised. The effect of the genetic variation at the APOE gene on biochemical variables was more pronounced in the HIV-infected individuals who carried the apoE2/3 genotype. The highly active anti-retroviral therapy (HAART)-receiving group presented increased levels of total cholesterol and apoE. Dyslipidemia was a predictable consequence of HIV infection and the protease inhibitors intensified the increase in apoE values.

Fungos em péletes de laranja e alergia respiratória em trabalhadores da indústria citrícola / Association between fungi from orange pellets and respiratory allergy in workers of citrus plant

Objetivos: Na indústria citrícola, a casca e o bagaçoda laranja são prensados, macerados e desidratados em um forno a 800ºC, convertendo-se em grãos denominados péletes. Observamos que o seu manuseio parecia estar relacionado com alergia respiratória, o que suscitou essa pesquisa com o objetivo de determinar a prevalência desses sintomas na amostra estudada e esclarecer a natureza do(s) alérgeno(s) envolvido(s). Métodos: Estudo realizado na indústria e pátios de caminhões onde foram coletadas amostras de péletes e foram isolados fungos segundo técnicas micológicas padronizadas. Operários e caminhoneiros (n=56) foram submetidos a questionário, testes cutâneos de leitura imediata (extratos Alk-Abelló) e coleta de sangue (dosagem de IgE específica por quimioluminescência). Resultados: Em 57 amostras de péletes, 86 per cent apresentavam crescimento de fungos, predominantemente dos gêneros: Cladosporium, Rhizopus, Aspergillus, Trichoderma e Penicillium. Encontrou-se 35,7 per cent de sintomáticos respiratórios.A positividade de testes cutâneospara fungos (38,5 per cent), entre a população de caminhoneiros sintomáticos foi significativa (p<0,015).As dosagens de IgE específicas corroboraram as evidênciasclínicas, uma vez que 92,3 per cent dos soros testados de caminhoneiros com sintomas respiratórios de alergia foram positivos para fungos, contrapondo-se aos grupos controles cuja positividade foi de 7,4 per cent (p< 0,001). Conclusões: Caminhoneiros expostos a aerodispersíveis provenientes de péletes da laranja podem desenvolver alergia respiratória por mecanismo de hipersensibilidade tipo I, induzida por fungos, que contaminam este material.