Amazonian medicinal plants efficiently inactivate Herpes and Chikungunya viruses.

The Amazonian species investigated in this research are commonly utilized for their anti-inflammatory properties and their potential against various diseases. However, there is a lack of scientifically supported information validating their biological activities. In this study, a total of seventeen ethanolic or aqueous extracts derived from eight Amazonian medicinal plants were evaluated for their activity against Herpes Simplex type 1 (HSV-1) and Chikungunya viruses (CHIKV). Cytotoxicity was assessed using the sulforhodamine B method, and the antiviral potential was determined through a plaque number reduction assay. Virucidal tests were conducted according to EN 14476 standards for the most potent extracts. Additionally, the chemical composition of the most active extracts was investigated. Notably, the LMLE10, LMBA11, MEBE13, and VABE17 extracts exhibited significant activity against CHIKV and the non-acyclovir-resistant strain of HSV-1 (KOS) (SI > 9). The MEBE13 extract demonstrated unique inhibition against the acyclovir-resistant strain of HSV-1 (29-R). Virucidal assays indicated a higher level of virucidal activity compared to their antiviral activity. Moreover, the virucidal capacity of the most active extracts was sustained when tested in the presence of protein solutions against HSV-1 (KOS). In the application of EN 14476 against HSV-1 (KOS), the LMBA11 extract achieved a 99.9% inhibition rate, while the VABE17 extract reached a 90% inhibition rate. This study contributes to the understanding of medicinal species native to the Brazilian Amazon, revealing their potential in combating viral infections that have plagued humanity for centuries (HSV-1) or currently lack specific therapeutic interventions (CHIKV).

Metabolomic analysis among ten traditional "Arnica" (Asteraceae) from Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of several Asteraceae species in Brazil are popularly used as anti-inflammatory. Some of these species are popularly recognizes as "arnica" because of the morphological and sensorial analogy with the traditional European Arnica montana. These used species in Brazil were identified as Calea uniflora Less, Chaptalia nutans (L.) Polák, Lychnophora ericoides Mart. Lychnophora pinaster Mart. Lychnophora salicifolia Mart. Porophyllum ruderale (Jacq.) Cass, Pseudobrickellia brasiliensis (Spreng.) R. M. King & H. Rob. Sphagneticola trilobata (L.) Pruski and Solidago chilensis Meyen. However, the comparative chemical profile of these so-called "arnicas" has never been reported in the literature. AIM OF THE STUDY: This work aimed to compare the main plants recognized as "arnica" in Brazil by using metabolomic analysis, based on UPLC-ESI-QTof-MS2 data and multivariate statistical analysis. MATERIALS AND METHODS: The metabolites profiling of 10 "arnica" species were established by UPLC-ESI-QTof-MS2. Three tinctures of each species (dry leaves) were produced and one aliquot of each tincture was injected and analyzed three times by UPLC-ESI-QTof-MS2. Data were acquired both in negative and positive modes and processed by MassLynx®, MarkerLynx® and Matlab® softwares. Principal component analysis (PCA) was used to reduce dimensionality and data redundancy; hierarchical trees helped to identify and eliminate contaminated or misplaced injections/samples. To achieve the objectives both hierarchical and k-means clustering techniques were employed to group similar samples or species. RESULTS: Diagnostic analysis of MS data allowed the identification of 54 metabolites. The identification was supported with the use of an external standard, fragmentation pattern and data from the literature. The main classes of identified compounds included phenolic acids, coumarin, flavonoids, heterosides, terpenoids and nitrogen compounds. Cluster analysis revealed that Sphagneticola trilobata, Solidago chilensis and Lychnophora pinaster have some chemical features similar to those of Arnica montana. In contrast, the same statistical analysis also showed that Pseudobrickellia brasiliensis, Porophyllum ruderale and Chaptalia nutans are chemically diverse from Arnica montana. The variability of the samples relied principally on nitrogenated compounds (confidence level 4) found in P. brasiliensis and P. ruderale, three phenolic compounds (level 2) detected in P. brasiliensis and in C. nutans and triterpenes (level 3) found in L. salicifolia and L. pinaster. CONCLUSIONS: In summary, the mass spectrometry technique in conjunction with multivariate statistical analysis proved to be an excellent tool to identify correlated compounds, as well as to verify the chemical similarity among evaluated species. This methodology was successfully used to establish important correlations in medicinal preparations of so-called "arnicas" used in Brazil.

Effects of Standardized Medicinal Plant Extracts on Drug Metabolism Mediated by CYP3A4 and CYP2D6 Enzymes.

The use of medicinal plants concomitantly with conventional drugs can result in herb-drug interactions that cause fluctuations in drug bioavailability and consequent therapeutic failure and/or toxic effects. The CYP superfamily of enzymes plays an important role in herb-drug interactions. Among CYP enzymes, CYP3A4 and CYP2D6 are the most relevant since they metabolize about 50% and 30% of the drugs on the market, respectively. Thus, the main goal of this study was to evaluate the occurrence of in vitro interactions between medicinal plant extracts and drug substrates of CYP3A4 and CYP2D6 enzymes. Standardized extracts from nine medicinal plants (Bauhinia forficata, Cecropia glaziovii, Cimicifuga racemosa, Cynara scolymus, Echinacea sp., Ginkgo biloba, Glycine max, Ilex paraguariensis, and Matricaria recutita) were evaluated for their potential interactions mediated by CYP3A4 and CYP2D6 enzymes. Among the extracts tested, C. glaziovii (red embaúba) showed the most relevant inhibitory effects of CYP3A4 and CYP2D6 activity, while I. paraguariensis (yerba mate) inhibited CYP3A4 activity. Both extracts were chemically analyzed by UPLC-MS/MS, and these inhibitory effects could lead to clinically potential and relevant interactions with the drug substrates of these isoenzymes.

Antiprotozoal investigation of 20 plant metabolites on Trypanosoma cruzi and Leishmania amazonensis amastigotes. Atalantoflavone alters the mitochondrial membrane potential.

The study aims to evaluate the antiprotozoal activities of 20 plant metabolites on Trypanosoma cruzi and Leishmania amazonensis amastigotes. Compounds 1-20 were obtained and identified by using chromatographic and spectroscopic techniques. The antiparasitic assays were performed on the intracellular form of T. cruzi and L. amazonensis using human leukaemic THP-1 cells as the host. The mechanism of action of the most active compounds was explored in silico by molecular docking using T. cruzi trypanothione reductase (TR) as a target, whereas the in vitro studies were performed by enzymatic assay using T. cruzi recombinant TR. In addition, the mitochondrial membrane potential was evaluated by flow cytometry. Two flavonoids, one triterpene and three acetogenins showed from high to moderate trypanocidal activities with IC50 values ranging 3.6-37.2 µm while three of the metabolites were moderately leishmanicidal. The molecular docking study revealed interactions between TR and the most trypanocidal compounds 1 (abyssinone IV) and 2 (atalantoflavone). In contrast, both showed no effect on TR in vitro. For the mitochondrial membrane potential assay, atalantoflavone (2) displayed a dose-dependent depolarization. On the basis of the aforementioned results, this compound's structure could be chemically explored in order to develop more potent trypanocidal derivatives.

Estrogenic and cytotoxic potentials of compounds isolated from Millettia macrophylla Benth (Fabaceae): towards a better understanding of its underlying mechanisms.

BACKGROUND: Millettia macrophylla was previously reported to have estrogenic effects and to prevent postmenopausal osteoporosis in Wistar rats. So, the study deals with the identification of its secondary metabolites and the evaluation of their estrogenicity and cytotoxicity toward tumoural cells. Thus, 13 known compounds were obtained from successive chromatographic columns and identified by NMR data compared to those previously reported. METHODS: In vitro estrogenicity of the isolates and the phenolic fraction (PF) of M. macrophylla were performed by E-screen and reporter gene assays, while their cytotoxicity was evaluated by Alamar Blue (resazurin) assay. A 3-days uterotrophic assay and the ability of PF to alleviate hot flushes in ovariectomized adult rats were tested in vivo. RESULTS: Seven of the 13 secondary metabolites turned to be estrogenic. Only two exhibited cytotoxic effects on MCF-7 and MDA-MB-231 with CC50 values of 110 µM and 160 µM, respectively. PF induced a significant (p < 0.01) MCF-7 cells proliferation and transactivated both ERα and ERß in the reported gene assay at 10-2 µg/mL. In vivo, PF acted more efficiently than the methanol crude extract, resulting to a significant (p < 0.01) increase in the uterine wet weight, uterine protein level, uterine and vaginal epithelial height at the dose of 10 mg/kg BW. In addition, PF reduced the average duration and frequency of hot flushes induced in rat. CONCLUSION: These aforementioned results indicate that PF is a good candidate for the preparation of an improved traditional medicine able to alleviate some menopausal complaints such as vaginal dryness and hot flushes. Estrogenic and cytotoxic potentials of compounds isolated from Millettia macrophylla Benth. (Fabaceae): towards a better understanding of its underlying mechanism.

Crateva adansonii DC, an African ethnomedicinal plant, exerts cytotoxicity in vitro and prevents experimental mammary tumorigenesis in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Crateva adansonii DC is a plant traditionally used in Cameroon to treat constipation, asthma, snakebites, postmenopausal complaints and cancers. AIM: The anticancer potential of the dichloromethane/methanol extract of C. adansonii stem barks was investigated using human breast cancer cell and 7,12 dimethylbenz(a)anththracene (DMBA)-induced mammary tumorigenesis model in rats. MATERIAL AND METHODS: The cytotoxicity of C. adansonii extract was assessed in vitro towards breast carcinoma (MCF-7 and MDA-MB-231) and non-tumoral cell lines (NIH/3T3 and HUVEC) by Alamar Blue assay. Furthermore, in vivo studies were performed on female Wistar rats treated either with C. adansonii extract at a dose of 75 or 300mg/kg body weight or with tamoxifen (3.3mg/kg body weight), starting 1 week prior DMBA treatment and lasted 12 weeks. The investigation focused on tumour burden, tumour DNA fingerprint, morphological, histological, hematological, and biochemical parameters. RESULTS: CC50 values for the in vitro assays were 289µg/mL against MCF-7 cells and >500µg/mL in others cells, leading to a selectivity index ≥1.73. C. adansonii extract significantly (p<0.001) revealed in vivo the reduction of the cumulative tumour yield (87.23%), total tumour burden (88.64%), average tumour weight (71.11%) and tumour volume (78.07%) at the dose of 75mg/kg as compared to DMBA control group. A weak effect was also observed at 300mg/kg. This extract showed a moderate hyperplasia at the dose of 75mg/kg while at 300mg/kg no significant change was noted as compared to DMBA group. It protected rats from the DNA alteration induced by DMBA and increased antioxydant enzymes activities in mammary gland tissue homogenates. In addition, Ultra-High Performance Liquid Chromatography/ESI-QTOF-Mass Spectrometry analysis of C. adansonii extract detected structure-related of many well-known anticancer agents such as flavane gallate, flavonol, phenylpropanoïds, sesquiterpene derivatives, gallotannins and lignans. The LD50 of C. adansonii was estimated to be greater than 5000mg/kg. CONCLUSIONS: These aforementioned results suggest that the C. adansonii extract may possess antitumor constituents, which could combat breast cancer and prevent chemically-induced breast cancer in rats.

Herbalists, traditional healers and pharmacists: a view of the tuberculosis in Ghana

This paper is the result of a visit by Brazilian researchers to Ghana, with the aim of improving understanding of the relationship between traditional healers and conventional health practices, specifically in relation to tuberculosis. Through this exploratory visit, this group of researchers promoted by the Edital Pro Africa (CNPq) had an opportunity to learn about, reflect on, and discuss the different social, economic and cultural realities and contexts that have led to the different health conditions and forms of healthcare in Ghana. Besides the direct relationship between the social and economic conditions of the country and the health of its population, it was also concluded that there is a clear distancing, in the Ghanaian reality, between the traditional healers and the conventional system, in terms of culture and modes of operation, each constituting isolated systems with little or no collaboration between them.The visit enabled us to see the difficulties involved in managing TB, including diagnosis, treatment, monitoring and co-infection with HIV. The majority of patients with TB only go to hospital after several attempts at self-medication, due to the non-specificity of the principal symptoms, and also to the trust in the traditional medicine. Initiatives to encourage research into medicinal plants in Ghana are seeking partnerships with developed countries, but not always with clear or secure national interests. For the traditional healers, there are high hopes that the information gathered by researchers from the local universities, on the plants and traditional methods they use, will result in affirmation and recognition of their practices, but they complain strongly that they receive no feedback on the research carried out.

A new fatty aldol ester from the aerial part of Mimosa invisa (Mimosaceae).

A new aldol ester named 17-O-triacontanoylheptadecanal (1) was isolated from the aerial part of Mimosa invisa (Mimosaceae) together with eight known compounds identified as ß-sitosterol (2), α-amyrine (3), lupeol (4), 4'-O-methylepinumisoflavone (5), alpinumisoflavone (6), betulinic acid (7), 3-O-ß-D-glucopyranoside of sitosterol (8) and epirobinetinidol (9). The structures of compounds were determined on the basis of NMR and mass spectrometry data as well as by comparing the data reported in the literatures. The antimicrobial activities of the crude extract and compounds 1 and 9 were investigated against seven microbial species. The natural products showed moderate activities compared to that of the crude extract.