ABSTRACT Objective: To determine the prevalence and factors associated with herbal medicinal product use among primary healthcare patients in southeastern Brazil. Methods: A cross-sectional study combined with documentary research was conducted between August and November 2017, involving interviews with 1,221 adults following dispensing from primary healthcare units. The characterization of herbal medicinal product usage included assessing the prevalence, redundancy, and interaction indicators, as well as identifying sources of recommendation and acquisition. Descriptive statistics were used for variable analysis and logistic regression was used to estimate the factors associated with herbal medicinal product usage, calculating odds ratios at a significance level of 5%. Results: The prevalence of herbal medicinal product usage was 3.8%, with 60% of them prescribed by physicians. Private pharmacies (53%) were the main sources, and Valeriana officinalis (42%) was the most cited plant species and the sole herbal medicinal product on the municipal list. The indicators of redundancy and interactions between concurrently prescribed pharmaceutical and herbal medicinal products were 42.9% and 28.6%, respectively. Herbal medicinal products usage correlated with the female sex (OR= 2.50; 95%CI= 1.11-5.59), non-white race (OR= 0.52; 95%CI= 0.28-0.94), monthly income above 1 minimum wage (OR= 3.48; 95%CI= 1.51-8.01), and self-reported of anxiety, mood disorder or depression (OR= 2.97; 95%CI= 1.55-5.66). Conclusion: Despite the low use of herbal medicinal products, users face a high potential for interactions and therapeutic redundancy. Special attention should be paid to patients taking herbal medicinal products and allopathic medicines concurrently. Women with anxiety, mood disorders, or depression were more likely to use herbal medicinal products, whereas non-white people were 48% less likely to use them, suggesting potential socioeconomic disparities in access. Objective Brazil Methods crosssectional cross sectional 2017 1221 221 1,22 units acquisition 5 5% Results 38 3 8 3.8% 60 physicians 53% 53 (53% 42% 42 (42% list 429 9 42.9 286 28 6 28.6% respectively OR= OR (OR 2.50 250 2 50 95CI CI 95 95%CI 1.115.59, 111559 1.11 5.59 , 11 59 1.11-5.59) nonwhite non white 0.52 052 0 52 0.280.94, 028094 0.28 0.94 94 0.28-0.94) 3.48 348 48 1.518.01, 151801 1.51 8.01 51 01 1.51-8.01) selfreported self reported anxiety 2.97 297 97 1.555.66. 155566 1.55 5.66 . 55 66 1.55-5.66) Conclusion disorders access 201 122 22 1,2 3.8 (53 4 (42 42. 28.6 2.5 25 115 1.115.59 11155 111 1.1 559 5.5 1.11-5.59 0.5 05 280 0.280.94 02809 028 0.2 094 0.9 0.28-0.94 3.4 34 518 1.518.01 15180 151 1.5 801 8.0 1.51-8.01 2.9 29 555 1.555.66 15556 155 566 5.6 1.55-5.66 20 12 1, 3. (5 (4 28. 2. 1.115.5 1115 1. 5. 1.11-5.5 0. 0.280.9 0280 02 09 0.28-0.9 1.518.0 1518 15 80 8. 1.51-8.0 1.555.6 1555 56 1.55-5.6 ( 1.115. 1.11-5. 0.280. 0.28-0. 1.518. 1.51-8. 1.555. 1.55-5. 1.115 1.11-5 0.280 0.28-0 1.518 1.51-8 1.555 1.55-5 1.11- 0.28- 1.51- 1.55-

BACKGROUND In a screen of extracts from plants and fungi to detect antileishmanial activity, we found that the ethyl acetate extract of the fungus Nectria pseudotrichia, isolated from the tree Caesalpinia echinata (Brazilwood), is a promising source of bioactive compounds. OBJECTIVES The aims of this study were to isolate and determine the chemical structures of the compounds responsible for the antileishmanial activity of the organic extract from N. pseudotrichia. METHODS Compounds were isolated by chromatographic fractionation using semi-preparative high-performance liquid chromatography, and their chemical structures were determined by analytical and spectral data and by comparison with published data. The antileishmanial activity of the isolated compounds was evaluated in intracellular amastigote forms of Leishmania (Viannia) braziliensis expressing firefly luciferase as reporter gene, and cytotoxicity was determined in Vero and THP-1 mammalian cell lines by MTT assay. FINDINGS Fractionation of the extract yielded seven compounds: 10-acetyl trichoderonic acid A (1), 6′-acetoxy-piliformic acid (2), 5′,6′-dehydropiliformic acid (3), piliformic acid (4), hydroheptelidic acid (5), xylaric acid D (6), and cytochalasin D (7). Compounds 1, 2 and 3 are reported here for the first time. Compounds 1, 2, and 5 were more active, with IC50 values of 21.4, 28.3, and 24.8 µM, respectively, and showed low toxicity to Vero and THP-1 cells. MAIN CONCLUSIONS N. pseudotrichia produces secondary metabolites that are more toxic to intracellular amastigote forms of L. (V.) braziliensis than to mammalian cells.

Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL), Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a concentration of 20 μg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular culture assay.

Organic extracts from leaves and stems of Stillingia oppositifolia Baill. ex Müll. Arg., Euphorbiaceae, were screened for antifungal and cytotoxic properties. The extracts presented Minimum Inhibitory Concentration values around 250 µg.mL-1 against Candida krusei and Candida tropicalis, and around 63 µg.mL-1 for Paracoccidioides brasiliensis. They were tested on three human cell lines (UACC-62, MCF-7, and TK-10), disclosing GI50 values, (concentration able to inhibit 50% of the cell growth) ranging from 50 to 100 µg.mL-1. Organic extract from stems furnished hexanic, dichloromethanic and aqueous phases after partition. Chromatographic fractionation of the hexanic soluble phase of the stems yielded aleuritolic acid 3-acetate, β-sitosterol, 3-epi-β-amyrin, β-amyrone and palmitic acid. These compounds showed antifungal and cytotoxic activities in the same range as the organic crude extract and low toxic effect against mononuclear cells obtained from human peripheral blood. This is the first report on chemical and biological potential of S. oppositifolia.

The fungus Lentinus strigosus (Pegler 1983) (Polyporaceae, basidiomycete) was selected in a screen for inhibitory activity on Trypanosoma cruzi trypanothione reductase (TR). The crude extract of L. strigosus was able to completely inhibit TR at 20 µg/ml. Two triquinane sesquiterpenoids (dihydrohypnophilin and hypnophilin), in addition to two panepoxydol derivatives (neopanepoxydol and panepoxydone), were isolated using a bioassay-guided fractionation protocol. Hypnophilin and panepoxydone displayed IC50 values of 0.8 and 38.9 µM in the TR assay, respectively, while the other two compounds were inactive. The activity of hypnophilin was confirmed in a secondary assay with the intracellular amastigote forms of T. cruzi, in which it presented an IC50 value of 2.5 µ M. Quantitative flow cytometry experiments demonstrated that hypnophilin at 4 µM also reduced the proliferation of human peripheral blood monocluear cells (PBMC) stimulated with phytohemaglutinin, without any apparent interference on the viability of lymphocytes and monocytes. As the host immune response plays a pivotal role in the adverse events triggered by antigen release during treatment with trypanocidal drugs, the ability of hypnophilin to kill the intracellular forms of T. cruzi while modulating human PBMC proliferation suggests that this terpenoid may be a promising prototype for the development of new chemotherapeutical agents for Chagas disease.