Investigations on the morpho-anatomy and histochemistry of Aconitum napellus L.

Aconitum napellus L. is a popular medicinal plant extensively used in homeopathy. This article provides detailed morphology and microscopy, including the anatomical and histochemical features of the herb, to aid authentication and quality control. In cross-section, the root in secondary growth shows the phloem surrounded by pericyclic fibers and a well-developed xylem. The stem is irregular in outline, displaying unicellular trichomes and many free collateral vascular bundles encircling the pith. The leaf is dorsiventral, hypostomatic with anomocytic and anisocytic stomata, and shows non-glandular trichomes. The floral parts are characterized by uniseriate epidermises, homogeneous mesophyll, anomocytic stomata on the abaxial surface, trichomes, and oval pollen grains. The tissue fragments in powdered herbs show these characteristics and have numerous starch grains with thimble-shaped, linear or star-shaped hilum. The detailed macroscopic and microscopic analysis provided in this study can help in the authentication and quality control of A. napellus raw materials. RESEARCH HIGHLIGHTS: Key anatomical, micromorphological, and microchemical features of Aconitum napellus are described. The results of the study can support the taxonomy of the genus Aconitum. Morphological standardization of the species reported here is helpful in the quality control of this herb.

A Scoping Review of Genus Viscum: Biological and Chemical Aspects of Alcoholic Extracts.

The genus Viscum comprises a large number of semi-parasitic shrubs popularly known as Mistletoe. The Viscum species grow in many countries of Europe, Africa and Asia with different popular uses in ornamentation, foods and medicine. Many studies about Viscum have been done over the last years focusing on biological activities and chemical composition of the aqueous extracts, mainly related to anthroposophical medicines. However, it is known that non-aqueous preparations, as alcoholic extracts, have demonstrated different biological activities that are species-and host tree-dependent. Considering the potential of these alcoholic extracts, a scoping review was conducted using data from three online databases: PubMed, Scopus and Embase. Inclusion criteria consisted of the in vitro, in vivo, ex vivo, clinical and chemical studies of alcoholic extracts from Viscum species. The present review summarized 124 original publications about fourteen Viscum species. Viscum album, Viscum articulatum and Viscum coloratum were the main studied species. Alcoholic extracts demonstrated hypotensive, anticancer, antimicrobial, analgesic and anti-inflammatory capabilities, among other biological activities. Flavonoids, phenolic acids and terpenoids represented 48%, 24% and 11% of the total identified compounds, respectively. This review contributes to the knowledge of alcoholic preparations of the Viscum species and points out the lack of clinical studies concerning these different extracts.

Viscum album mother tinctures: Harvest conditions and host trees influence the plant metabolome and the glycolytic pathway of breast cancer cells.

Viscum album is a semi-parasitic plant used for over one hundred years in complementary cancer therapy. The main commercial drugs used in cancer patients' treatment are derived from the aqueous V. album extracts, whose cytotoxic potential is mostly attributed to the aqueous soluble antitumoral metabolites. On the counterpart, ethanol solvents must be used to obtain V. album mother tinctures. This methodology permits better solubilization of phenolic compounds, among others, which present antitumoral bioactivity. Recently, the metabolomics approach revealed the influence of the host tree on the V. album subspecies differentiation. To increase the scientific information about the chemical differences related to the host trees and to clarify the seasonal influences, in this study, the metabolome of 50 V. album mother tinctures from three subspecies (abietis, album, austriacum) and five host trees (Malus domestica, Quercus sp., Ulmus carpinifolia, Pinus sylvestris, Abies alba) was evaluated using summer and winter plant harvests. The in vitro cytotoxic activities were investigated in breast cancer cells (MDA-MB-231) and immortalized normal human keratinocytes (HaCaT). The summer V. album mother tinctures presented higher cytotoxic activity than winter ones. Among the summer samples, those prepared with V. album subsp. album were more cytotoxic than V. album subsp. abietis and subsp. V. album subsp. austriacum. The V. album harvested from Quercus petraea and Abies alba inhibited the key-glycolytic enzymes: hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK). This activity was related to a reduction in glucose uptake and lactate production, which were host-tree-time-dose-dependent. The untargeted metabolomic approach was able to discriminate the mother tinctures according to respective botanical classes and harvest season. A total of 188 metabolites were annotated under positive and negative modes. Fourteen compounds were responsible for the samples differentiation, and, to the best of our knowledge, eight were described in the Viscum album species for the first time. Our study shows the interruption of the Warburg effect as a novel antitumoral mechanism triggered by V. album mother tinctures, which is related to their metabolite profile. These results bring scientific evidence that encourages the use of V. album mother tinctures as a natural product for cancer therapy.

NMR analysis: potential and challenges as a homeopathic basic research tool

Homeopathic ultra-diluted solutions surpass the threshold of matter dispersion and, consequently, their chemical constitution is similar to inert solvent. Nevertheless, randomized clinical trials have shown that the clinical effects of these homeopathic medicines are superior to placebo1. Nuclear magnetic resonance (NMR) is one of the most promising techniques to detect physicochemical alterations induced by homeopathic procedures2,3. Aims: To investigate T2 NMR relaxation times of Zincum metallicumand lactose dynamized samples. Methodology: Zincum metallicumsamples were ground until 6dH using lactose as an excipient. Subsequently, these samples were dynamized with ultrapure water to produce 8dH, 9dH, 10dH,and 11dH. Lactose dynamized samples (6dH-11dH) were used as control. Aliquots of 540µl of each sample were diluted with 60µl of deuterated water (D2O) in 5mm tubes. The analyses were carried out in Bruker Ascend TM 500MHZ spectrometer at 288 K. Results and discussion: The Zincum metallicumand lactose T2 relaxation times were very similar, except for Zincum metallicum8dH, which presented a value of 1.226 in comparison to 1.036 of lactose 8dH. The following T2 values were registered: 1.287 -9dH; 1.413 -10dH; 1.467 -11dH, and 1.303 -9dH; 1.400 ­10dH; 1.350 -11dH, for Zincum metallicumand lactose, respectively. The differences detected in 8dH samples are probably due to the presence of lactose in the first dilution step, in which 1 part of the 6dH triturated mixture was diluted in 9 parts of water, to prepare 7dH. Following this homeopathic procedure, 8dH solutions remain around 1% of lactose which could be influenced by the T2 values registered.Conclusion: These preliminary results showed the possibility to apply the NMR technique to evaluate the influence of dynamization in the relaxation parameters. Further studies should be carried out with other potencies and/or other homeopathic substances, in addition to the evaluation of T1 and the T1/T2 parameters, as previously described by other groups.

Viscumalbum L. homeopathic mother tinctures: metabolome and antitumor activity

Viscum album L. is a semi-parasitic plant with antitumor activity attributed to theaqueous extracts. However, European V. album ethanolic extracts (VAE) have also demonstrated invitro activity in tumor models. Aims: Evaluate the metabolic profiles of fifty VAE harvested duringsummer and winter seasons and their antitumor activity through 2D and 3D models. Methodology:VAEwerepreparedbymacerationfrom:V.albumsubsp.albumgrowingonMalus domestica,Quercus sp.and Ulmus sp.; V. album subsp. austriacum from Pinus sylvestris; V. album subsp. abietis from Abies alba.Chemical analyses were performed through liquid chromatography coupled with high resolutionmass spectrometry and Partial Least Squares Discriminant Analysis (PLS-DA) was performed in theMetaboanalyst 4.0. The antitumor potential of the selected VAE was evaluated in 2D and 3D models(MDA-MB-231 cancer cells) by MTT, crystal violet and glycolytic pathway analysis. Results anddiscussion:Thefirst3principalcomponentsinPLS-DAexplained60%and40%ofdatavariationin positive andnegativemodesrespectively.Threegroupswereformedandshowedchemicalsimilarityamong V. album subspecies. The compounds responsible for group separation were tentativelyidentifiedas:pinobankasinornaringenin hexoside;isorhamnetin-3-hexoside,meglutolanddifferent aminoacids.ThesummerVAEat0.5%v/vinducedhighercytotoxicdamagethanthewinterpreparations, and Abies alba and Quercus sp. VAE promoted 49% and 42% reduction of tumorviability in 3D model (72h incubation), respectively. MDA-MB-231 glycolytic pathway in 2D modelshowed a decrease in the glucose consumption and extracellular lactate production. Also, PFK (6-phosphofructo-1-kinase)andPK(Pyruvatekinase)activitieswereinhibitedbyAbiesalbaandQuercus sp. VAE at 48h of incubation. Conclusion: VAE extracts showed different metabolomes andthe glycolytic pathway should be an important target involved in the inhibition of tumor growth bytheseextracts

The use of homeopathy and flower therapy as an aid in the treatment of climacteric symptoms: an integrative review

Climacteric is characterized as the set of symptoms that usually start in the period of transition from the reproductive phase to female senility, and directly impacts woman's quality of life. Currently, the treatments approved by the FDA for this pathology mainly involve the use of antidepressants and hormone replacement, both having side effects. Clinical studies carried out in 2002, showed an increased risk of breast cancer, and other pathologies related to the prolonged use of these drugs. The impact ofthe studies resulted in a greater interest in complementary and alternative medicines (CAMs), such as the use of homeopathy and flower remedies for the treatment of climacteric symptoms. This abstract aims to present an integrative review on the use of homeopathy and flower remedies in the treatment of climacteric symptoms, in order to identify it main scientific evidence. For this, metodology consisted of research in the databases Web of Science, Google Scholar, HomeoIndex, LILACS and SciELO. The inclusion criteria were original human studies, totally available, and published in the last 10 years in Portuguese, English and Spanish, presenting the use of floral remedies or homeopathic treatment. The main homeopathic medicines used were Lachesis mutus, Belladona, Sepia officinalis, Sanguinaria canadensis and Sulfur. The main flower remedies were Cherry plum, Agrimony, Gentian, Walnut, Olive and Larch. The findings in this review demonstrate a positive outcome trend in favor of the effectiveness of these practices through the studies evaluated, encouraging the expansion of new designs and research that fully contemplate the principles of these practices. Thus, this work contributes to the advancement in the understanding of each one of these rationalities, making this work a source of consultation for health professionals and for future research, resulting in a strengthening of CAMs in the field of health.

Heteroisotherapics effect on the treatment against smoking: a cognitive-behavioral approach

The most common way of consuming nicotine is in tobacco cigarettes. Nicotine causes intense addiction. The National Cancer Institute coordinates and executes the Tobacco Control Program in Brazil, through actions that encourage the adoption of healthier lifestyles. In this context, homeopathy has used Heteroisotherapic medicines formulated according to the homeopathic pharmaceutical technology with scientific evidence of efficacy in the detoxification of substances and metals, and in the desensitization of foods or medicines. Aims Promote the importance of the cognitive-behavioral approach in combination with the homeopathic treatment against smoking. Methodology In the initial phase of the randomized double-blind clinical study (CEP / HUCFF / UFRJ 65622916.2.0000.5257), the effectiveness of the 6CH heteroisotherapeutic drug was assessed. Volunteers were recruited andin-person welcoming meetings, using the cognitive-behavioral approach, were carried out to inform them about the risks of smoking and the benefits of quitting. In addition, they were supported and guided during the smoking cessation process so that they could deal with the withdrawal syndrome, the psychological dependence and the constraints associated with smoking. Results and discussion84 participants were selected according to the inclusion criteria, and divided by randomization into two groups:the Test Group (heteroisotherapeutic medication) and the Control Group (homeopathic medication Nux vomica6CH).Both groups will be followed for 12 months. The combination of the following approaches has led to a significant increase in the cessation rate: I.Prepare the smoker for solving his own issues; II. Stimulate skills to resist temptations to smoke; III. Prepare to prevent relapse; IV. Prepare to deal with stress. Studies show that, regardless the duration of these approaches, there is an increase in the abstinence rate. Moreover, the longer the total approach time (frequency multiplied by the time spent on each contact), the higher the abstinence rate. On the other hand, from a total approach time of 90 minutes on, there is no further increase in the abstinence rate. ConclusionThe partial results obtained so far demonstrate that the cognitive-behavioral approach played a decisive role in the groups performance, favoring the treatment adherence as well as the group cohesion around the Project's objective, contributing to the effectiveness of the medicine, a decreased anxiety, improved sleep, cessation or decrease in the number of cigarettes smoked per day and the abstinence rate.

Medicines for the new coronavirus in the view of Classical Systemic Homeopathy.

INTRODUCTION: According to the World Health Organization, the coronavirus disease 2019 (COVID-19) has 48.896.564 reported cases and 1.236.995 deaths worldwide on November 5, 2020. Despite great efforts, there is no treatment of COVID-19 up to nowadays. Therefore, it is essential to search for therapeutic alternatives to COVID-19, such as Integrative Medicine, especially Homeopathy, which was used with excellent results in the great epidemics. AIM: This study aims to describe the coronavirus Pandemic from the perspective of Classical Systemic Homeopathy and to identify the homeopathic medicine (medicines genius) that has its sphere of action in most of the symptoms involved in this pandemic. METHODS: Articles published in indexed journals and websites of medical institutions, books of homeopathic materia medica, scientific journals, and government publications were reviewed. RESULTS: A total of 39 references were selected and enclosed 20 articles indexed on COVID-19, 08 references of systemic conditions, 11 articles indexed of homeopathy, 06 classic homeopathy books. After the study, the homeopathic medicine Cinchona officinalis (China officinalis) was proposed, according to pathogenesis (experimental pathophysiological study in humans) described in the materia medica books consulted, for the relief of symptoms. Its sphere of action in most of the symptoms involved in this pandemic. Moreover, acute homeopathic medicines were determined to act in the relief of symptoms of the various phases of the manifestations of the disease. CONCLUSION: Data contributes to use in homeopathic interventions during the COVID-19 epidemic as a health promotion and treatment strategy that can be used as an adjunct to all sanitary and therapeutic measures recommended by health authorities.

Metabolomics by UHPLC-Q-TOF Reveals Host Tree-Dependent Phytochemical Variation in Viscum album L.

Viscum album L., commonly known as European mistletoe, is a hemi-parasitic plant of the Santalaceae family. The in vitro and in vivo effects of V. album differ, according to its host tree. However, little is known about the host-dependent phytochemical diversity in V. album. In this study, the metabolic profiles of V. album ssp. album from Malus domestica Bork., Quercus robur L., and Ulmus carpinifolia Gled were compared. Leaves, stems, and berries were collected in Switzerland, by the same procedure, in September 2016 and 2017. The methanolic extracts were analyzed by ultra-performance liquid chromatography, coupled to electrospray quadrupole time-of-flight mass spectrometry in positive ionization mode. The data were submitted to partial-least square discriminant analysis (PLS-DA) and the results showed that the V. album ssp. album samples were clustered into three groups, according to the three distinct host trees. Seven compounds, with high VIP scores (variable importance in projection), were responsible for this differentiation. The following four compounds were detected in both the harvest years: arginine, pipecolic acid or lysine, dimethoxycoumarin, and sinapyl alcohol, suggesting their use as host specific V. album biomarkers. The present work highlights the importance of standardized harvest and analytical procedures for the reproducibility of the chemical results of herbal materials.

Phytochemical analysis and in vitro anti-proliferative activity of Viscum album ethanolic extracts.

BACKGROUND: Viscum album L. (Santalaceae), commonly known as mistletoe, is a hemiparasitic plant traditionally used in complementary cancer treatment. Its antitumor potential is mostly attributed to the presence of aqueous soluble metabolites; however, the use of ethanol as solvent also permits the extraction of pharmacological compounds with antitumor potential. The clinical efficacy of mistletoe therapy inspired the present work, which focuses on ethanolic extracts (V. album "mother tinctures", MT) prepared from different host trees. METHODS: Samples from three European subspecies (album, austriacum, and abietis) were harvested, and five different V. album-MT strains were prepared. The following phytochemical analyses were performed: thin layer chromatography (TLC), high-performance liquid chromatography (HPLC) and liquid chromatography-high resolution mass spectrometry (LC-HRMS). The proliferation assay was performed with WST-1 after incubation of tumor (Yoshida and Molt-4) and fibroblast cell lines (NIH/3 T3) with different MT concentrations (0.5 to 0.05% v/v). The cell death mechanism was investigated by flow cytometry (FACS) using Annexin V-7AAD. RESULTS: Chemical analyses of MT showed the presence of phenolic acids, flavonoids and lignans. The MT flavonoid and viscotoxin contents (mg/g fresh weight) were highest in Quercus robur (9.67 ± 0.85 mg/g) and Malus domestica (3.95 ± 0.58 mg/mg), respectively. The viscotoxin isoform proportions (% total) were also different among the VA subspecies with a higher content of A3 in V. album growing on Abies alba (60.57 ± 2.13). The phytochemical compounds as well as the viscotoxin contents are probably related to the antitumor effects of MT. The cell death mechanisms evaluated by colorimetric and FACS methodologies involved necrotic damage, which was host tree-, time- and dose- dependent, with different selectivity to tumor cells. Mother tincture from V. album ssp. abietis was the most effective at inducing in vitro cellular effects, even when incubated at the smallest concentration tested, probably because of the higher content of VT A3. CONCLUSION: Our results indicate the promising antitumor potential of Viscum album ethanolic extracts and the importance of botanical and phytochemical characterization for in vitro anti-proliferative effects.

Mapa de Evidência: efetividade clínica da Fitoterapia e Plantas Medicinais para Cicatrização e Doenças Agudas / Evidence Map: clinical effectiveness of Medicinal Plants and Phytotherapy for Wound Healing and Acute Diseases / Mapa de Evidencia: efetividad clínica de las Plantas Medicinales y Fitoterapia para Cicatrización y Enfermedades Agudas

O mapa apresenta uma visão geral das evidências sobre os efeitos das Plantas Medicinais e Fitoterapia para cicatrização e doenças agudas. A partir de uma ampla busca bibliográfica foram incluídos no mapa 16 estudos de revisão sistemática. Todos os estudos foram avaliados, caracterizados e categorizados por um grupo de pesquisadores do Laboratório de Pesquisa & Desenvolvimento de Práticas Integrativas e Complementares (LabPICs) da Faculdade de Farmácia da Universidade Federal do Rio de Janeiro.

El mapa presenta una visión general de la evidencia sobre los efectos de las plantas medicinales y la fitoterapia para cicatrización y las enfermedades agudas. A partir de una amplia búsqueda bibliográfica, se incluyeron 16 estudios de revisión sistemática en el mapa. Todos los estudios fueron evaluados, caracterizados y categorizados por un grupo de investigadores del Laboratorio de Investigación y Desarrollo para Prácticas Integrativas y Complementarias (LabPIC) en la Facultad de Farmacia de la Universidad Federal de Rio de Janeiro.

The map presents an overview of the evidence on the effects of Medicinal Plants and Phytotherapy for for wound healing and acute diseases. From a wide bibliographic search, 16 systematic review studies were included in the map. All studies were evaluated, characterized and categorized by a group of researchers from the Research & Development Laboratory for Integrative and Complementary Practices (LabPICs) at the Faculty of Pharmacy, Federal University of Rio de Janeiro.

Mapa de Evidência: efetividade clínica das Plantas Medicinais e Fitoterapia para Saúde Mental e Qualidade de Vida / Mapa de Evidencia: efetividad clínica de las Plantas Medicinales y Fitoterapia para Salud Mental y Calidad de Vida / Evidence Map: clinical effectiveness of Medicinal Plants and Phytotherapy for Mental Health and Quality of Life

O mapa apresenta uma visão geral das evidências sobre os efeitos das Plantas Medicinais e Fitoterapia para Saúde Mental e Qualidade de Vida. A partir de uma ampla busca bibliográfica foram incluídos no mapa 37 estudos de revisão sistemática. Todos os estudos foram avaliados, caracterizados e categorizados por um grupo de pesquisadores do Laboratório de Pesquisa & Desenvolvimento de Práticas Integrativas e Complementares (LabPICs) da Faculdade de Farmácia da Universidade Federal do Rio de Janeiro.

El mapa presenta una visión general de la evidencia sobre los efectos de las plantas medicinales y la fitoterapia para ela salud mental y calidad de vida. A partir de una amplia búsqueda bibliográfica, se incluyeron 37 estudios de revisión sistemática en el mapa. Todos los estudios fueron evaluados, caracterizados y categorizados por un grupo de investigadores del Laboratorio de Investigación y Desarrollo para Prácticas Integrativas y Complementarias (LabPIC) en la Facultad de Farmacia de la Universidad Federal de Rio de Janeiro.

The map presents an overview of the evidence on the effects of Medicinal Plants and Phytotherapy for mental health and quality of life. From a wide bibliographic search, 37 systematic review studies were included in the map. All studies were evaluated, characterized and categorized by a group of researchers from the Research & Development Laboratory for Integrative and Complementary Practices (LabPICs) at the Faculty of Pharmacy, Federal University of Rio de Janeiro.

Mapa de Evidência: efetividade clínica das Plantas Medicinais e Fitoterapia para Distúrbios Metabólicos e Fisiológicos / Mapa de Evidencia: efetividad clínica de las Plantas Medicinales y Fitoterapia para Trastornos Metabólicos y Fisiológicos Clinical / Evidence Map: effectiveness of Medicinal Plants and Phytotherapy for Metabolism and Physiological Disorders

O mapa apresenta uma visão geral das evidências sobre os efeitos das Plantas Medicinais e Fitoterapia para Distúrbios Metabólicos e Fisiológicos. A partir de uma ampla busca bibliográfica foram incluídos no mapa 48 estudos de revisão sistemática. Todos os estudos foram avaliados, caracterizados e categorizados por um grupo de pesquisadores do Laboratório de Pesquisa & Desenvolvimento de Práticas Integrativas e Complementares (LabPICs) da Faculdade de Farmácia da Universidade Federal do Rio de Janeiro.

El mapa presenta una visión general de la evidencia sobre los efectos de las plantas medicinales y la fitoterapia para el trastornos metabólicos y fisiológicos. A partir de una amplia búsqueda bibliográfica, se incluyeron 48 estudios de revisión sistemática en el mapa. Todos los estudios fueron evaluados, caracterizados y categorizados por un grupo de investigadores del Laboratorio de Investigación y Desarrollo para Prácticas Integrativas y Complementarias (LabPIC) en la Facultad de Farmacia de la Universidad Federal de Rio de Janeiro.

The map presents an overview of the evidence on the effects of Medicinal Plants and Phytotherapy for Metabolic and Physiological Disorders. From a wide bibliographic search, 48 systematic review studies were included in the map. All studies were evaluated, characterized and categorized by a group of researchers from the Research & Development Laboratory for Integrative and Complementary Practices (LabPICs) at the Faculty of Pharmacy, Federal University of Rio de Janeiro.

Mapa de Evidência: efetividade clínica das Plantas Medicinais e Fitoterapia para Doenças Crônicas e Dor / Mapa de Evidencia: efetividad clínica de las Plantas Medicinales y Fitoterapia para Enfermedades Cronicas y Dolor / Evidence Map: clinical effectiveness of Medicinal Plants and Phytotherapy for Chronic Diseases and Pain

O mapa apresenta uma visão geral das evidências sobre os efeitos das Plantas Medicinais e Fitoterapia para dor e doenças crônicas. A partir de uma ampla busca bibliográfica foram incluídos no mapa 49 estudos de revisão sistemática. Todos os estudos foram avaliados, caracterizados e categorizados por um grupo de pesquisadores do Laboratório de Pesquisa & Desenvolvimento de Práticas Integrativas e Complementares (LabPICs) da Faculdade de Farmácia da Universidade Federal do Rio de Janeiro.

El mapa presenta una visión general de la evidencia sobre los efectos de las plantas medicinales y la fitoterapia para el dolor y las enfermedades crónicas. A partir de una amplia búsqueda bibliográfica, se incluyeron 49 estudios de revisión sistemática en el mapa. Todos los estudios fueron evaluados, caracterizados y categorizados por un grupo de investigadores del Laboratorio de Investigación y Desarrollo para Prácticas Integrativas y Complementarias (LabPIC) en la Facultad de Farmacia de la Universidad Federal de Rio de Janeiro.

The map presents an overview of the evidence on the effects of Medicinal Plants and Phytotherapy for pain and chronic diseases. From a wide bibliographic search, 49 systematic review studies were included in the map. All studies were evaluated, characterized and categorized by a group of researchers from the Research & Development Laboratory for Integrative and Complementary Practices (LabPICs) at the Faculty of Pharmacy, Federal University of Rio de Janeiro.

Protective effect of Myrsine parvifolia plant extract against the inflammatory process induced by Bothrops jararaca snake venom.

Accidents involving snakes from the genus Bothrops sp. constitute the most important cause of snake envenomation in Brazil. The Myrsine genus has been reported to be used in folk medicine against snakebites. In this work, the phytochemical profiles and ability of extracts from Myrsine parvifolia leaves to reduce the inflammatory process (edema, vascular permeability increase and leukocyte migration) induced by Bothrops jararaca venom were investigated in vivo. Chemical compounds were identified by chromatographic and spectroscopy techniques. Total polyphenol, tannin, and flavonoid contents were determined by spectrophotometric methods. Swiss male mice received an oral administration of extracts (100 mg/kg) in different protocols. Paw edema, intraperitoneal vascular permeability and pleurisy models in mice were used to evaluate the antiophidic potential of the extracts. Paw edema was induced by subplantar injection of B. jararaca venom and quantified as the increase in paw volume. Changes in vascular permeability were assessed by measuring the amount of Evans blue dye extravasation. Leukocyte migration was assessed by total and differential counts in the pleural cavity washes. Myricetin, myricetin-3-O-ß-arabinopyranoside, quercetin and kaempferol were isolated from the ethyl acetate extract and identified as the primary compounds of the dichloromethane extract. Terpenes and fatty acids were identified in the hexane and dichloromethane extracts. The pretreated group with hydroethanolic and dichloromethane extract reduced total edema (40 and 52%, respectively), vascular permeability increase (32.4 and 32.2%, respectively) and leukocyte influx into the pleural cavity (42 and 39%, respectively), while the group treated with hexane extract showed only reduced edema (37%) induced by B. jararaca venom. The hydroethanolic extract showed better results in all of the tests performed and was also administered by the protocol of post-poisoning, showing maintenance of paw edema reduction and cell migration. These data indicate a potential anti-inflammatory activity of M. parvifolia in poisoning by B. jararaca, especially to reduce local poison effects.

Phenolic compounds from Viscum album tinctures enhanced antitumor activity in melanoma murine cancer cells.

Cancer is one of the biggest problems in public health worldwide. Plants have been shown important role in anticancer research. Viscum album L. (Santalaceae), commonly known as mistletoe, is a semi-parasitic plant that grows on different host trees. In complementary medicine, extracts from European mistletoe (Viscum album L.) have been used in the treatment of cancer. The study was conducted to identify chemical composition and antitumor potential of Viscum album tinctures. Chemical analysis performed by high resolution chromatography equipped with high resolution mass spectrometer identified caffeic acid, chlorogenic acid, sakuranetin, isosakuranetin, syringenin 4-O-glucoside, syringenin 4-O-apiosyl-glucoside, alangilignoside C and ligalbumoside A compounds. Some of these compounds are probably responsible for the reduction of tumoral cellular growth in a dose-dependent manner. It was observed that melanoma murine cells (B16F10) were more sensitive to V. album tinctures than human leukaemic cells (K562), besides non-tumoral cells (MA-104) had a much lower cytotoxicity to them. Apoptotic-like cells were observed under light microscopy and were confirmed by a typical DNA fragmentation pattern. Additionally, flow cytometry results using Annexin-V/FITC permitted to quantify increased expression of early and late apoptotic markers on tumoral cells, confirming augmented Sub G0 population, which was probably associated with a consistent decrease in G1, and an increase in S or G2/M populations. Results indicate the chemical composition of V. album tinctures influences the mechanisms of in vitro tumoral cell death, suggesting a potential use in cancer pharmacotherapy research.

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.