Direct electric current modifies important cellular aspects and ultrastructure features of Candida albicans yeasts: Influence of doses and polarities.

Available treatments against human fungal pathogens present high levels of resistance, motivating the development of new antifungal therapies. In this context, the present work aimed to analyze direct electric current (DC) antifungal action, using an in vitro apparatus equipped with platinum electrodes. Candida albicans yeast cells were submitted to three distinct conditions of DC treatment (anodic flow-AF; electroionic flow-EIF; and cathodic flow-CF), as well as different charges, ranging from 0.03 to 2.40 C. Our results indicated C. albicans presented distinct sensibility depending on the DC intensity and polarity applied. Both the colony-forming unit assay and the cytometry flow with propidium iodide indicated a drastic reduction on cellular viability after AF treatment with 0.15 C, while CF- and EIF-treated cells stayed alive when DC doses were increased up to 2.40 C. Additionally, transmission electron microscopy revealed important ultrastructural alterations in AF-treated yeasts, including cell structure disorganization, ruptures in plasmatic membrane, and cytoplasmic rarefaction. This work emphasizes the importance of physical parameters (polarity and doses) in cellular damage, and brings new evidence for using electrotherapy to treat C. albicans pathology process. Bioelectromagnetics. 38:95-108, 2017. © 2016 Wiley Periodicals, Inc.

Structural and thermal analyses of zinc and lactose in homeopathic triturated systems.

BACKGROUND: A series of different experimental approaches was applied in Zincum metallicum (Zinc met.) samples and lactose controls. Experiments were designed to elucidate the effect of zinc trituration and dynamization on physicochemical properties of homeopathic formulations, using lactose as excipient. METHODS: Zinc met. potencies (Zinc met 1-3c) were triturated and dynamized using lactose as excipient, according to Brazilian Homeopathic Pharmacopoeia. Lactose samples (LAC 1-3c) were also prepared following the same protocol and used as controls. The samples were analyzed structurally by Atomic Absorption Spectroscopy (AAS), X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM) with Energy Dispersive X-ray Spectroscopy (EDX) and Scanning Electron Microscopy (SEM), and thermodynamically by Thermogravimetry (TG) and Differential Scanning Calorimetry (DSC). RESULTS: AAS analysis detected 97.0 % of zinc in the raw material, 0.75 % (Zinc met 1c) and 0.02% (Zinc met 2c). XRD analysis showed that inter-atomic crystalline spacing of lactose was not modified by dynamization. Amorphous and crystalline lactose spheres and particles, respectively, were observed by TEM in all samples, with mean size from 200 to 800 nm. EDX obtained with TEM identified zinc presence throughout the amorphous matter but individualized zinc particles were not observed. SEM images obtained from dynamized samples (LAC 1c and Zinc met 1c) with electron backscattering could not identify zinc metal grains. The dynamization process induced Derivatives of Thermal Gravimetric (DTg) peak modification, which was previously centered near 158°C to lactose, to a range from 140 to 170°C, suggesting the dynamization process modifies the temperature range of water aggregation. Thermal phenomena were analyzed and visualized by Analysis of Variance (ANOVA) and Principal Component Analysis (PCA) statistics. Both indicated that fusion enthalpy of dynamized samples (DynLAC 1-3c; DynZn 1-3c) increased 30.68 J/g in comparison to non-dynamized lactose (LAC; p < 0.05). CONCLUSIONS: Our results suggested no structural changes due to the trituration and dynamization process. However, TG and DSC analyses permit the differentiation of dynamized and non-dynamized groups, suggesting the dynamization process induced a significant increase in the degradation heat. These results call for further calorimetric studies with other homeopathic dilutions and other methodologies, to better understand the dynamics of these systems.

Effects of direct electric current on Herpetomonas samuelpessoai: An ultrastructural study.

The literature shows that the effects of direct electric currents on biological material are numerous, including bactericidal, fungicidal, parasiticidal, and anti-tumoral, among others. Non-pathogenic trypanosomatids, such as Herpetomonas samuelpessoai, have emerged as important models for the study of basic biological processes performed by a eukaryotic cell. The present study reports a dose-dependent anti-protozoan effect of direct electric treatment with both cathodic and anodic current flows on H. samuelpessoai cells. The damaging effects can be attributable to the electrolysis products generated during electric stimulation. The pH of the cell suspension was progressively augmented from 7.4 to 10.5 after the cathodic treatment. In contrast, the anodic treatment caused a pH decrease varying from 7.4 to 6.5. Transmission electron microscopy analyses revealed profound alterations in vital cellular structures (e.g., mitochondrion, kinetoplast, flagellum, flagellar pocket, nucleus, and plasma membrane) after exposure to both cathodic and anodic current flows. Specifically, cathodic current flow treatment induced the appearance of autophagic-like structures on parasite cells, while those submitted to an anodic current flow presented marked disorganization of plasma membrane and necrotic appearance. However, parasites treated in the intermediary chamber (without contact with the electrodes) did not present significant changes in viability or morphology, and no pH variation was detected in this system. The use of H. samuelpessoai as a biological model and the direct electric current experimental approach used in our study provide important information for understanding the mechanisms involved in the cytotoxic effects of this physical agent.

Acupuntura no alívio da dor osteomuscular e a melhora na funcionalidade na pessoa idosa: estudo quase experimental / Acupuncture in the relief of osteomuscular pain and improvement of functionality in older adults: a quasi-experimental study

Resumo Objetivo Avaliar os efeitos da acupuntura no alívio da dor osteomuscular em pessoas idosas e a possibilidade de melhora da funcionalidade para as atividades básicas de vida diária. Método Estudo quase experimental, com avaliação antes e depois da intervenção utilizando a Escala Visual Analógica e o Índice de Katz. Amostra por conveniência, constituída de 31 pessoas idosas. O período de intervenção comportou quatro sessões de atendimentos semanais. A intervenção com acupuntura abordou um protocolo de tratamento sistêmico por intermédio de cinco pares de pontos: IG4 (Hegu), F3 (Taichong), E36 (Zusanli), PC6 (Neiguan) e BP6 (Sanyinjiao). Resultados 100% (31) informando sentir dor, sendo 64,52% (20) em intensidade moderada (3-7); 25,81% (8) dor intensa (8-10); e 9,68% (3) dor leve (0-2). O protocolo de tratamento mostrou-se positivo em 4 sessões, com redução da dor ao grau leve, e média geral reduzida abaixo de 2 no score. A análise dos resultados da Escala Visual Analógica e os do índice de Katz, mostrou possibilidades do uso da acupuntura no alívio da dor osteomuscular e na melhora da funcionalidade da pessoa idosa. Conclusão O uso da técnica de acupuntura sistêmica apontou resultados satisfatórios no alívio da dor osteomuscular e na melhoria da funcionalidade para as atividades básicas de vida diária da pessoa idosa.

Abstract Objective To assess the effects of acupuncture on the relief of osteomuscular pain in older adults and the potential improvement in functionality for activities of daily living. Method A quasi-experimental study, with pre- and post-intervention assessment using the Visual Analog Scale and the Katz Index. Convenience sample comprising 31 older adults. The intervention period consisted of four weekly treatment sessions. Acupuncture intervention targeted a systemic treatment protocol utilizing five pairs of acupoints: LI4 (Hegu), LIV3 (Taichong), ST36 (Zusanli), PC6 (Neiguan), and SP6 (Sanyinjiao). Results 100% (31) reported experiencing pain, with 64.52% (20) reporting moderate pain (3-7), 25.81% (8) reporting severe pain (8-10), and 9.68% (3) reporting mild pain (0-2). The treatment protocol demonstrated positive outcomes after 4 sessions, with a reduction of pain to a mild degree and an overall average score reduced below 2. The analysis of results from the Visual Analog Scale and the Katz Index showed potential for the use of acupuncture in relieving osteomuscular pain and improving the functionality of the older adult. Conclusion The use of systemic acupuncture techniques yielded satisfactory results in the relief of osteomuscular pain and the enhancement of functionality for activities of daily living of the older adult.

Comparative analysis for quality control of mother tinctures in Brazilian reports

Quality control of ahomeopathic mother tincture begins with the analysis of the raw material. Once the product is obtained, some organoleptic characteristics and Physicochemical properties such as appearance, colour, odour, density, dry residue, alcohol content,and chemicalmarker are evaluated. In Brazil, mother tinctures from different suppliers may have high variability in terms of the specifications described in the homeopathic pharmacopoeia. Objective: The aim of the present study was to compare mother tinctures' quality control from different pharmaceutical suppliers based on Brazilian Homeopathic Pharmacopoeia 3rd edition. Methods: Five mother tinctures were selected according to the high number of requests in the pharmacies, also its monographs are in the Brazilian Homeopathic Pharmacopoeia (FHB) 3rd edition. The report of analysis of the product was provided by three homeopathic pharmacies from the city of Rio de Janeiro. The information in the reports wasevaluated and compared to its monograph. Results and discussion: In this study, it was found that the number of laboratories that provide homeopathic pharmaceutical ingredients is rather low. In addition, in some reports, a number of identification tests werenot described properly when compared to the monograph. Besides that, some information diverges from the pharmacopoeia, for example, absence of dry residue testing, replacement of the part of the plant used to prepare the mother tincture,and results that do not meet pharmacopoeia specifications. As can be seen, the quality and effectiveness of homeopathic medicines prepared from these tinctures may be compromised. Conclusion:In order to guarantee consumer protection and the quality and effectiveness of medicines, governments and regulatory agencies should requirea greater commitment from suppliers in the production of homeopathic inputs.

Encapsulation of Piper cabralanum (Piperaceae) nonpolar extract in poly(methyl methacrylate) by miniemulsion and evaluation of increase in the effectiveness of antileukemic activity in K562 cells.

This study aimed to synthesize and characterize nanoparticles (NPs) of poly(methyl methacrylate) (PMMA) and evaluate their ability to incorporate plant extracts with antitumor activity and low dissolution in aqueous media. The extract used was n-hexane partition of the methanol extract of Piper cabralanum (PCA-HEX). PMMA NPs were obtained using the mini-emulsion method, which was able to encapsulate almost 100% of PCA-HEX. The synthesized polymeric particles presented with a size of 200 nm and a negative charge. Cytotoxicity tests by MTT and trypan blue assays showed that NPs without PCA-HEX did not kill leukemic cells (K562 cells). NPs containing PCA-HEX were able to enhance cell death when compared to pure extract. The results showed that PMMA NPs could be useful as a drug delivery system as they can enhance the antitumor activity of the PCA-HEX extract by more than 20-fold. PMMA NPs containing plant extracts with antitumor activities may be an alternative to control the evolution of diseases such as leukemia.

In vitro evaluation of Sporothrix brasiliensis biotherapic

Sporothrix brasiliensisis one of themost virulent zoonosis which affects animals and humans. This fungus is responsible for subcutaneous infection and its contamination is possible through trauma to the skin. Sporotrichosis is highly prevalent in feline. And Rio de Janeiro appears to have the highest occurrence of cases. Objectives: This study aims to evaluate the in vitroefficacy of Sporothrix brasiliensisbiotherapic, with and without an association to allopathic medicine commonly used in the treatment.Methodology: Conidiumcells of Sporothrix brasiliensiswill be cultured in Potato dextrose agar (PDA) for 5 to 7 days and yeast cells in Brain heart infusion (BHI) for 3 to 5 days. After incubation, the cells will be scraped with a drigalski handle and filtered using cells strainer into a tube and centrifuge for 5 minutes at 3000 RPM. The cells will be resuspended with Phosphate buffered saline (PBS), centrifuge again,and finally resuspended in PBS. After preparing the inocule, the microplates will be prepared. There will be 5groups in vitro. The first one will be the control group, only fungi. The second will be the treatment of fungi with homeopathic medicine (Sporothrix brasiliensis30DH). The third group will be the homeopathic medicine in association with itraconazole. The fourth will be the treatment with itraconazole only. And the last group will be the fungi with dynamized distilled water 30DH. Sporothrix brasiliensis30DHwill be prepared according to Brazilian Homeopathic Pharmacopeia. Results and discussions: The experiments are still in progress and the results will be analyzed through Analysis of variance (ANOVA) to determine statistically significant differences. Previous articles based on biotherapic treatments demonstrated successful results, so our research group is conducting these experiments to evaluate the effect in this model. Conclusion: Experiments will be made to verify the efficacy of biotherapic on sporotrichosis treatments.

Direct electric current treatment modifies mitochondrial function and lipid body content in the A549 cancer cell line.

Electrochemical therapy (EChT) entails treatment of solid tumors with direct electric current (DC). This work evaluated the specific effects of anodic flow generated by DC on biochemical and metabolic features of the A549 human lung cancer cell line. Apoptosis was evaluated on the basis of caspase-3 activity and mitochondrial transmembrane potential dissipation. Cell morphology was analyzed using transmission electron microscopy, and lipid droplets were studied through morphometric analysis and X-ray qualitative elemental microanalysis. High-resolution respirometry was used to assess mitochondrial respiratory parameters. Results indicated A549 viability decreased in a dose-dependent manner with a prominent drop between 18 and 24h after treatment (p<0.001), together with a two-fold increase in caspase-3 activity. AF-treatment induced a significantly increase (p<0.01) in the cell number with disrupted mitochondrial transmembrane potential. Furthermore, treated cells demonstrated important ultrastructural mitochondria damage and a three-fold increase in the cytoplasmic lipid bodies' number, quantified by morphometrical analyses. Conversely, 24h after treatment, the cells presented a two-fold increase of residual oxygen consumption, accounting for 45.3% of basal oxygen consumption. These results show remarkable alterations promoted by anodic flow on human lung cancer cells which are possibly involved with the antitumoral effects of EChT.

Nanostructured delivery system for zinc phthalocyanine: preparation, characterization, and phototoxicity study against human lung adenocarcinoma A549 cells.

In this study, zinc phthalocyanine (ZnPc) was loaded onto poly-ɛ-caprolactone (PCL) nanoparticles (NPs) using a solvent emulsification-evaporation method. The process yield and encapsulation efficiency were 74.2% ± 1.2% and 67.1% ± 0.9%, respectively. The NPs had a mean diameter of 187.4 ± 2.1 nm, narrow distribution size with a polydispersity index of 0.096 ± 0.004, zeta potential of -4.85 ± 0.21 mV, and spherical shape. ZnPc has sustained release, following Higuchi's kinetics. The photobiological activity of the ZnPc-loaded NPs was evaluated on human lung adenocarcinoma A549 cells. Cells were incubated with free ZnPc or ZnPc-loaded NPs for 4 h and then washed with phosphate-buffered saline. Culture medium was added to the wells containing the cells. Finally, the cells were exposed to red light (660 nm) with a light dose of 100 J/cm(2). The cellular viability was determined after 24 h of incubation. ZnPc-loaded NPs and free photosensitizer eliminated about 95.9% ± 1.8% and 28.7% ± 2.2% of A549 cells, respectively. The phototoxicity was time dependent up to 4 h and concentration dependent at 0-5 µg ZnPc. The cells viability decreased with the increase of the light dose in the range of 10-100 J/cm(2). Intense lysis was observed in the cells incubated with the ZnPcloaded NPs and irradiated with red light. ZnPc-loaded PCL NPs are the release systems that promise photodynamic therapy use.

Bioterápicos de Candida albicans induzem alterações nos índices de adesão e na respiração celular / Biotherapics of Candida albicans induce changes in the levels of adhesion and in cellular respiration

Introdução: O gênero Candida spp é responsável por cerca de 80% das infecções fúngicas no ambiente hospitalar e constitui causa relevante de infecções sistêmicas em pacientes hospitalizados, especialmente em doentes graves e em imunocomprometidos, com predominância da Candida albicans. A adesão das leveduras às células epiteliais do hospedeiro é um potente estimulador para a formação de hifas, forma invasiva do fungo [1]. Os bioterápicos são medicamentos preparados a partir de produtos biológicos, elaborados conforme a Farmacopeia Homeopática Brasileira (FHB)[2], indicados para tratamento de infecções de etiologia conhecida, empregados com grande sucesso no tratamento clínico destas infecções. Os bioterápicos RC, desenvolvidos pelo médico brasileiro Roberto Costa (RC) são preparados a partir do agente etiológico íntegro e, segundo suas pesquisas, possuem maior capacidade de estimular o sistema imunológico do hospedeiro.

High dilutions of Euphorbia tirucalli L. (aveloz) modify the viability and glycolytic metabolism of cell lines / Altas diluições de Euphorbia tirucalli L. (aveloz) modificam a viabilidade e o metabolismo glicolítico de linhas celulares

The latex of Euphorbia tirucalli L. (Aveloz) is popularly used to treat cancers from diverse sources such as: breast, prostate, lung and kidney. Furthermore, high dilutions of this latex (latex-HDs) have been successfully used in the treatment of tumors, although the mechanisms involved in this antitumoral activity are not yet known [...] Latex-HDs were obtained through the interaction of two procedures: 1:100 dilution in mass and agitation, using 2 solvents, water and 70% ethanol in homeopathic preparations 5, 15 and 30cH, following the Brazilian Homeopathic Pharmacopeia. Cell lines were incubated with 1 % and 10 % of latex-HDs for 24 hours. Controls treated with similar preparations without latex were incubated under the same conditions. Cell viability was analyzed by MTT assay and morphological features were assessed by May-Grunwald-Giemsa method. Cell ultrastructure was analyzed by transmission electron microscopy. Metabolic alterations were detected by spectrophotometric assay for the enzyme 6-phosphofructo-1-kinase (PFK-1) activity. 1% of ethanolic latex-HDs induced no alterations in MelanA cells viability, however Aveloz 15cH induced an increase in MCF7 cells viability. After aqueous treatment, MelanA cells viability decreased in almost all systems, whereas in MCF7 cells, an increase was detected in dynamized water systems and Aveloz 15CH. However, only water 15cH increased cell viability in comparison to control and non-dynamized water. No alterations in MCF7 morphology and ultrastructure were visualized through light microscopy or electronic microscopy, respectively. Interesting results were detected when we studied the glycolytic metabolism of both cells, giving support to evidences showing that HDs interfere in the metabolism of cell lines. Further studies are needed to better understand the mechanisms involved in this in vitro biological response to HDs.

O látex de Ephorbia tirucalli L. (aveloz) é usado popularmente no tratamento de diversos tipos de câncer incluindo mama, próstata, pulmão e rim. Além do mais, altas diluições desse látex (latex-HDs) têm sido utilizadas com sucesso no tratamento de tumores, embora o mecanismo dessa ação ainda seja desconhecido [...] As látex-HDs foram obtidas através de 2 procedimentos: diluição 1:100 em massa e agitação, com 2 solventes, água e etanol 70%, nas diluições homeopáticas 5, 15cH e 30cH de acordo com a Farmacopéia Homeopática Brasileira. As linhas celulares foram incubadas com látex-HDs 1% e 10% por 24 horas. Foram preparados controles similares sem o látex e incubados sob as mesmas condições. A viabilidade celular foi analisada através do método May-grunwald-Giemsa. A ultrastrutura celular foi analisada através de microscopia eletrônica de transmissão. As alterações metabólicas foram detectadas por ensaio de espectrofotometria para a atividade da enzima 6-fosfofructo-1-quinase (PFK-1). Látex-Hds em etanol 1% não induziram qualquer alteração na viabilidade das células MelanA, mas Aveloz 15cH induziu aumento da viabilidade das células MCF7. Após tratamento, a viabilidade das células MelanA diminuiu em quase todos os sistemas, enquanto que nas células MCF7 foi detectado aumento nos sistemas aquosos e Aveloz 15cH, mas só água 15cH aumentou a viabilidade celular por comparação ao controle e água não dinamizada. Não foram detectadas alterações na morfologia celular por microscopis de luz quando se estudou o metabolismo glicolítico em ambas as células, demonstrando que os padrões de resposta celular dependem do tipo de solvente, metabolismo celular e procedimento de diluição. Esta é a primeira vez que se mostra que as altas diluições interferem no metabolismo de linhas celulares. Outros estudos são necessários para se compreender melhor os mecanismos envolvidos nesta resposta in vitro a altas diluições de aveloz.

Chronic toxicological effects of high diluted solutions of Aveloz (Euphorbia tirucalli L. ) on healthy mice: a preliminary study / Efeitos toxicológicos crônicos de altas diluições de Aveloz (Euphorbia tirucalli L. ) em camundongos sadios: estudo preliminar

The latex extracted from Euphorbia tirucalli, a plant popularly known as Aveloz, is used in complementary medicine to induce tumor regression. However, as this latex has toxic effects when administered orally in ponderal doses, the present study was designed to assess the effects of high dilutions in healthy mice over a period of 18 weeks. The Aveloz latex-high diluted solutions (latex-HD) were obtained through the interaction of two processes: 1:100 dilution in mass and succussion, using ethanol 70% as a solvent, in the homeopathic dilutions of 5, 15 and 30cH, following Farmacopéia Homeopática Brasileira. Control solutions without latex were compounded (ethanolic-HD) in the same dilutions and were administered simultaneously. The animals which received latex-HD 30cH showed a significant increase in food consumption (p < 0.05) without significant difference in weight gain. In regards to water consumption, no statistical difference was shown when different Aveloz latex-HD groups were compared, apart from the group that received 1 drop of pure latex in water, which presented a significant increase (p < 0.05) in this parameter.

O látex extraído de Euphorbia tirucalli, planta conhecida popularmente como Aveloz, é utilizado em medicina complementar para induzir regressão tumoral. No entanto, como este látex tem efeitos tóxicos quando administrado por via oral em doses ponderais, este estudo foi desenhado para avaliar os efeitos de altas diluições do mesmo em camundongos sadios por um período de 18 semanas. As altas diluições do látex do Aveloz (látex-HD) foram obtidas através da interação de dois processos: diluição 1:100 em massa e sucussão, utilizando etanol 70% como solvente, nas diluições homeopáticas 5, 15 e 30cH, de acordo com a Famracopéia Homeopática Brasileira. Soluções de controle sem o látex (etanol-HD) foram compostas nas mesmas diluições e ministradas simultaneamente. Os animais que receberam látex-HD 30cH mostraram aumento significativo no consumo de alimento (p < 0,05) sem diferença significativa em ganho de peso. A respeito do consumo de água, não foram observadas diferenças significativas entre os diferentes grupos látex-HD, enquanto que o grupo que recebeu 1 gota de látex puro em água apresentou aumento significativo neste parâmetro (p < 0,05).