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This study evaluated the influence of Libidibia ferrea (Lf) extract used as dentin pretreatment on the resin-dentin bond strength stability and dentin endogenous enzymatic activity. The phytochemical profile (PP) of the Lf extract was evaluated by liquid chromatography; particle size, polydispersity index (PdI), and zeta potential (ZP) were evaluated by dynamic light scattering. The tested groups were ER-Scotchbond Universal (SBU) in the etch-and-rinse (ER) mode; ERLf-SBU in the ER mode + Lf after etching; SE- SBU in the self-etch (SE) mode; and LfSE-Lf before SBU in the SE mode. Sticks were obtained for microtensile bond strength tests and failure mode (24 hr and 12 months). The hybrid layer was evaluated using scanning electron microscopy. The endogenous enzymatic activity of the underlying dentin was analyzed by in situ zymography with the same treatments. The PP showed the presence of quercetin (2.6% w/w). Lf particles were considered large after the analysis of the PdI. The ZP remained stable over time. The ER and ERLf groups had lower bond strength after 12 months, but SE and LfSE remained stable. The predominant failure mode was adhesive for both times. ER and ERLf had longer resin tags and a thicker hybrid layer. The ER and LfSE groups showed higher enzymatic activity than the ERLf and SE groups after 12 months. The Lf extract may contribute to inhibit the dentin endogenous enzymatic activity when associated with an adhesive system in the ER mode.

Bisphosphonates are drugs used to treat bone disorders. The chronic use of bisphosphonates is associated with the occurrence of medication-related osteonecrosis of the jaw (MRONJ). Previous data reported the positive effects of Geranylgeraniol on different cell types treated with Bisphosphonates. Foregoing work done by our research group demonstrated the wound healing capacity of

Pterodon pubescens fruits are popularly used because of their analgesic and anti-inflammatory actions, which are attributed to the isolated compounds with a vouacapan skeleton. This work aimed to evaluate the antiproliferative and anti-inflammatory effects of a P. pubescens fruit dichloromethane extract and the vouacapan diterpene furan isomerʼs mixture (1 : 1) (6α-hydroxy-7β-acetoxy-vouacapan-17β-oate methyl ester and 6α-acetoxy-7β-hydroxy-vouacapan-17β-oate methyl ester isomers) in HaCaT cells using the cell migration and the BrDU incorporation assay. Levels of IL-8 were measured by ELISA after TNF-α stimulation. HPLC/DAD analysis of the extract revealed the expressive presence of vouacapan diterpene furan isomerʼs mixture. P. pubescens extract (1.5625 – 25 µg/mL) and vouacapan diterpene furan isomerʼs mixture (3.125 – 50 µM) inhibited cell proliferation as indicated by a decreased BrdU-incorporation. For the evaluation of cell migration, time-lapse microscopy was used. P. pubescens presented inhibition on cell migration at all concentrations tested (3.125 – 12.5 µg/mL), whereas for the VDFI mixture, the inhibition was only observed at the highest concentrations (12.5 and 25 µM) tested. Furthermore P. pubescens extract and vouacapan diterpene furan isomerʼs mixture significantly decreased IL-8 levels. Our results showed antiproliferative and anti-inflammatory effects on HaCaT cells treated with the extract and the vouacapan isomerʼs mixture, without affecting cell viability. These activities could be attributed to the voucapan molecular structures. In conclusion, topical products developed of P. pubescens extract or the voucapan isomerʼs mixture should be further studied as a potential product for local treatment against hyperproliferative lesions as in psoriasis vulgaris, representing an alternative treatment approach.

Aims: This study tested the antimicrobial effect of Arrabidaea chica (Ac) polyphenolic extract in different concentrations incorporated into an aqueous solution or into a primer of an adhesive system used for cavities pre-treatment against Streptococcus mutans (Sm) and Lactobacillus casei (Lc). Study Design: In vitro experimental study design to evaluate the minimum inhibitory concentration (MIC) of different solutions containing Ac against Sm and Lc. Methodology: Aqueous solution and primer (from a self-etching adhesive system - Clearfil SE Bond/Kuraray) were prepared containing Ac extract in six different concentrations: 0.0% (null); 0.25%; 0.5%; 1.0%; 1.5%; 2.0%; 2.5%. To determine the MIC, tubes with the inoculates of Sm and Lc were added with the solutions described above. Serial dilutions were inoculated in plates with culture medium (n=3). The colony forming unit count (CFU/mL) was carried out after 48 hours. Negative controls (aqueous or primer solutions without Ac incorporation) were used in both tests. Results: Regression model showed that all Ac concentrations inhibited Sm growth (p>0.05) in aqueous solution. Negligible or no Sm growth was observed in the presence of primer in Ac concentrations above 0.25% (p0.05). All concentrations of Ac tested here maintained the acidic characteristics of the solutions. Conclusion: Arrabidaea chica extract into an aqueous solution shows antimicrobial effect against Sm and Lc in all concentrations tested. When incorporated into the primer, the antimicrobial activity is observed in concentrations above 1%.

Pterodon pubescens Benth. is widely used in folk medicine for the treatment of inflammatory conditions, with the activity attributed to the compounds with a vouacapan moiety, however, few studies r...

Made available in DSpace on 2021-06-25T15:38:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-16 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Artemisinin (ART) was initially described for the control of inflammation and pain. However, the mechanisms involved with its antinociceptive effect are still poorly understood. Thus, this present study aimed to investigate the effect of ART in both free and nanocapsulated form on postoperative pain, as well as the participation of the spinal Toll-like receptor 4 (TLR4) in this process. Postoperative pain was induced using the skin/muscle incision retraction (SMIR) model in male Swiss mice. After 3 and 28 days of SMIR, the animals received an intrathecal injection of free or nanocapsulated ART, and the nociceptive threshold was evaluated by von Frey filament test. To evaluate the involvement of the microglia, astrocytes, and TLR4, minocycline (a microglia inhibitor), fluorocitrate (an astrocyte inhibitor), andLipopolysaccharide Rhodobacter sphaeroides(LPS-RS), a TLR4 antagonist, were intrathecally injected on the third day of SMIR. The levels of spinal TLR4 protein and proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), and interleukin-1-beta (IL-1 beta) were quantified by western blot and enzyme-linked immunosorbent assay, respectively. The results showed that free ART reduced postoperative pain (P < 0.001,F-5,F-30 = 7.49, 16.66% for 1000 ng dose; andP < 0.01,F-5,F-30 = 7.49, 14.58% for 500 ng dose) on the 3rd day of SMIR; while the ART nanocapsule had this effect on both the third (P < 0.001;F-5,F-30 = 4.94; 43.75, 39.58 and 72.91% for the 250, 500 and 1000 ng doses, respectively) and 28th (P < 0.05;F-5,F-30 = 7.71; 29.16 and 33.33% for the 500 and 1000 ng doses, respectively) day. The ART nanocapsule had a more potent and longer antinociceptive effect than free ART or morphine. Postoperative pain was also reduced by minocycline and LPS-RS. The ART nanocapsule also reduced the increased levels of TLR4, TNF-alpha, and IL-1 beta induced by SMIR. These data suggest that the ART nanocapsule has a potent analgesic effect on postoperative pain at the spinal level, and this response involves the inhibition of TLR4 and the proinflammatory cytokines TNF-alpha and IL-1 beta. Univ Fed Alfenas, Fac Motr Sci, Sci Motr Inst, Jovino Fernandes Sales Ave 2600, BR-37133840 Alfenas, Brazil Univ Sao Paulo, Dept Pharmacol, Sao Paulo, Brazil Univ Estadual Campinas, Fac Pharmaceut Sci, Campinas, Brazil Sao Paulo State Univ, Inst Sci & Technol, Sorocaba, Brazil Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Pharmacol, Ribeirao Preto, Brazil Sao Paulo State Univ, Inst Sci & Technol, Sorocaba, Brazil CAPES: 001 FAPESP: 2014/16008-3

Introduction Osteonecrosis of the jaw is a condition associated with intraoral ulceration and bone necrosis induced by antiresorptive medications, such as zoledronic acid, a bisphosphonate. Previous data on Arrabidaea chica (H&B.) Verlot wound healing activity prompted the study reported herein on A. chica standardized hydro alcoholic extract in vitro cytoprotective activity data on epithelial and osteoblastic cells exposed to zoledronic acid (ZA).

The tumor microenvironment presents several therapeutic targets, with inflammation being one of them. In search of new drugs, plants have shown to be an effective source of potent anti-inflammatory and anticancer agents. This study aimed to evaluate the antitumoral and inflammatory activities of Boehmeria caudata aerial parts extract. Bioguided in vitro antiproliferative screening showed that phenanthroquinolizidine obtained from the aerial B. caudata ethanolic extract had a straight relationship with activity. Moreover, the orally administered ethanolic extract reduced Ehrlich solid tumor growth and displayed an anti-inflammatory effect in both evaluated experimental models (carrageenan-induced paw edema and croton oil-induced ear edema). These results suggest that the antitumor activity of the ethanolic extract could be explained by antiproliferative effects associated with anti-inflammatory action.

Found in humid regions and waterways and popularly used to treat gastrointestinal problems among other applications, the present study evaluated the M. aquatica essential oil (OEMa) as a therapeutic alternative to treat gastrointestinal disorders. Produced by steam distillation, chemical composition of OEMa was determined by GC-MS analysis. The ethanol-induced ulcer and the dose-repeated acetylsalicylic acid (ASA)-induced gastrointestinal lesions models in rats evaluated, respectively, the prophylactic and curative effects of EOMa on peptic ulcers. The EOMa's effect on gastric secretion, gastric mucus and gastrointestinal motility were evaluated in in vivo models. The curative effect of EOMa on acute colitis was evaluated using the DSS-induced colitis model in mice. Obtained in 0.17% yield (w/w), with carvone (54.82 ± 1.39 g/100 g oil) as the main constituent, EOMa (at 75 mg/kg) showed potent gastroprotective effect ( 90%) mediated by non-protein sulfhydryl compounds (NPSH) and nitric oxide (NO) modulation alongside reduction in gastric secretion volume and total acidity. EOMa did not affect gastric mucus production and gastrointestinal motility. In dose-repeated ASA-induced gastrointestinal lesions model, EOMa (at 25 mg/kg) promoted the inflammatory process resolution both in gastric and duodenal walls by modulating NPSH, NO and myeloperoxidase levels. Despite delaying in 2 days the clinical symptoms worsening, EOMa (at 25 mg/kg) was not able to protect colon tissues from DSS-induced acute colitis as evidenced by macroscopic, biochemical, and histopathological parameters. This is the first report of Mentha aquatica essential oil as a promising herbal medicine for peptic ulcers treatment together with an adjuvant effect in IBD.

Bisphosphonates are drugs used to treat bone disorders. The chronic use of bisphosphonates is associated with the occurrence of medication-related osteonecrosis of the jaw (MRONJ). Previous data reported the positive effects of Geranylgeraniol on different cell types treated with Bisphosphonates. Foregoing work done by our research group demonstrated the wound healing capacity of Fridericia chica (Bonpl.) L.G.Lohmann standardized ethanol extract. Herein in vitro cytoprotective synergistic effect of the association of F. chica extract associated with an enriched geranylgeraniol fraction on keratinocytes exposed to zoledronic acid is reported. An association of F. chica at 1 and 5 µg/mL with geranylgeraniol at 15 µg/mL, increased cell viability by 73.5% and 71.1%, respectively. This treatment did not increase tumor cells viability; whereas the clonogenic potential assessment showed that, the association with F. chica (5 µg/mL) reversed the effects of zoledronic acid on the cells. This study provides data for a potential treatment for MRONJ.

Advanced wound dressings capable of interacting with lesions and changing the wound microenvironment to improve healing are promising to increase the therapeutic efficacy of this class of biomaterials. Aiming at the production of bioactive wound dressings with the ability to control the wound microenvironment, biomaterials of three different chemical compositions, but with the same architecture, were produced and compared. Electrospinning was employed to build up a biomimetic extracellular matrix (ECM) layer consisting of poly(caprolactone) (PCL), 50/50 dl -lactide/glycolide copolymer (PDLG) and poly( l -lactide) (PLLA). As a post-treatment to broaden the bioactivity of the dressings, an alginate coating was applied to sheathe and functionalize the surface of the hydrophobic electrospun wound dressings, in combination with the extract of the plant Arrabidaea chica Verlot, known for its anti-inflammatory and healing promotion properties. Wettable bioactive structures capable to interact with media simulating lesion microenvironments, with tensile strength and elongation at break ranging respectively from 155 to 273 MPa and from 0.94 to 1.39% were obtained. In simulated exudative microenvironment, water vapor transmission rate (WVTR) values around 700 g/m2/day were observed, while water vapor permeability rates (WVPR) reached about 300 g/m2/day. In simulated dehydrated microenvironment, values of WVTR around 200 g/m2/day and WVPR around 175 g/m2/day were attained.

Artemisia annua L. is the main artemisinin source, a sesquiterpene lactone used in the treatment against cancer and malaria, whose demand is high and the costs involving its production must be feasible. The main objective of the present work was to define a industrially and economically viable technology to obtain artemisinin. The experimental conditions of temperature, pressure and co-solvent use for obtaining extracts enriched in artemisinin by supercritical fluid extraction (SFE) were investigated, and an economic evaluation of the SFE was performed in comparison with the conventional extraction using ethanol. The techno-economic analysis showed that among the conditions evaluated in the present study, SFE at 60 °C and 250 bar, without co-solvent, was the most promising production condition. Nevertheless, for the commercial viability of the arteminisin extraction using SFE it is indicated to use A. annua rigorously selected, and to reduce costs of raw material.

Ursolic acid, a triterpene produced by plants, displayed leishmanicidal activity in vitro and in vivo, however, the low solubility of this triterpene limits its efficacy. To increase the activity of ursolic acid (UA), this triterpene was entrapped in nanostructured lipid carriers (UA-NLC), physical-chemical parameters were estimated, the toxicity was assayed in healthy golden hamsters, and the efficacy of UA-NLC was studied in experimental visceral leishmanisis. UA-NLC exhibited a spherical shape with a smooth surface with a size of 266 nm. UA-NLC displayed low polydispersity (PDI = 0.18) and good colloidal stability (−29.26 mV). Hamsters treated with UA-NLC did not present morphological changes in visceral organs, and the levels of AST, ALT, urea and creatinine were normal. Animals infected with Leishmania (Leishmania) infantum and treated with UA-NLC showed lower parasitism than the infected controls, animals treated with UA or Amphotericin B (AmB). The therapeutic activity of UA-NLC was associated with the increase in a protective immune response, and it was associated with a high degree of spleen and liver preservation, and the normalization of hepatic and renal functions. These data indicate that the use of lipid nanoparticles as UA carriers can be an interesting strategy for the treatment of leishmaniasis.

Bearing in mind the several medicinal properties of Mentha genus, this work aimed to evaluate the anti-proliferative potential of the ethanolic extract (EE) and fractions from M. aquatica L aerial parts. Using the anti-proliferative protocol developed by the NCI/USA, four fractions (F2 – F4 and F6) obtained from EE showed promising anti-proliferative profile against a panel of human tumor and non-tumor cell lines. After 24-h exposure, F2 (0.25 µg/mL) showed potent and irreversible anti-proliferative effect without inducing cell cycle arrest in both NCI-H460 and MCF-7 cells, without (anti) estrogenic activity. These effects were lost after storage of F2 diluted in dimethyl sulfoxide at −80 °C during 2 weeks. Analysis by gas chromatography coupled to mass detection evidenced some chemical changes induced by F2 storage in solution. The present study demonstrated the anti-proliferative effect of M. aquatica. Further studies are necessary to determine better storage conditions to enhance F2 stability.

Fridericia chica (Bonpl.) L.G. Lohmann (synonym Arrabidaea chica Verlot) is widely used in Brazilian folk medicine. Considering overcoming pitfalls of scaling up production of plant extracts, herein the effects of N2 atmosphere for extract spray-drying process is reported. Samples were monitored by in vitro antioxidant activity and microbiological evaluation. The drying atmosphere influenced 3-deoxyanthocyanines content when using air as atomizing gas, decreasing carajurin (37.5%) content with concomitant increase in luteolin yield (24.1%). Both drying processes preserved the pharmacological activity. In the cell migration test with HaCaT cells, the extract dried under air flow (5 μg/mL) promoted wound closure by 78% (12 hours) whereas the extract dried using N2 flow promoted 49% (12 hours), with 98% closure (12 hours) for the positive control. The antimicrobial evaluation for Staphylococcus aureus did not differ within drying atmospheres, with MIC (minimum inhibitory concentration) at 0.39 mg/mL. Therefore, the drying process reported herein did not interfere with the biological activity’s outcome., The authors A.L.T.G.R.and M.A.F thank CNPq for research productivity fellowship. The authors also thank the Chemical, Biological, and Agricultural Pluridisciplinary Research Center (CPQBA/Unicamp) for the laboratory infrastructure. Sao Paulo Research Foundation (FAPESP). This work was supported by the Coordination for the Improvement of Higher Education Personnel (CAPES, Brazil, Financing Code 001) and the National Council for Scientific and Technological Development (CNPq, Brazil, grant numbers # 132448/2016-5, # 132207/2017-6, # 429463/2018-9).

Malaria affects more than 200 million people annually around the world, killing a child every 2 min. Artemether (ART) and lumefantrine (LUM) are the gold standard choice to treat uncomplicated Plasmodium falciparum malaria; however, they are hydrophobic compounds with low oral bioavailability. Microneedle (MN) arrays consist of micron-sized needles on one side of a supporting base and have the ability to bypass the skin's stratum corneum barrier in a minimally invasive way, creating temporary channels through which drugs can diffuse, including those with poor water solubility. Herein, we report the development of dissolving MNs (DMNs) containing ART (MN-ART) and LUM (MN-LUM) as an alternative treatment regimen for malaria in low-resource settings. To incorporate the drugs into the MNs, nanosuspensions (NSs) for both molecules were developed separately to enhance drug solubility. The NSs were freeze-dried and the powder form was incorporated directly in an aqueous polymeric blend with poly-vinyl-pyrrolidone for MN-ART and a sodium hyaluronate hydrogel for MN-LUM. The in vivo bioavailability studies were performed using a MN reapplication scheme (1 × a day for 3 days), illustrating that an extended-release profile was achieved for both drugs when MNs were applied intradermally, and when compared to conventional oral treatment. The ART-LUM oral treatment was used as a positive control. For antimalarial activity, studies with animals infected with 106 Plasmodium yoelii 17XNL (12 days) were also conducted using female C57BL/6JUnib mice, demonstrating a 99.5% reduction in parasitemia by day 12 post-infection. By abolishing the infection, MN-ART and MN-LUM may serve as a promising controlled intradermal delivery device for antimalarial drugs to be explored in endemic areas.

Dental caries and periodontal disease are the most prevalent of the biofilm-dependent diseases. With numerous side effects on the use of chlorhexidine, the search for new safe therapeutic alternatives for microorganisms involved with these diseases increases every day. This study aimed to evaluate the antimicrobial activity and cytotoxicity of extracts made from the bark of Schinopsis brasiliensis Engl. against five oral microorganisms and analyze their phytochemical and thermal degradation profile. The liquid-liquid partition was performed with hexane, chloroform and ethyl acetate. The identification and quantification of the chemical marker was done. Antimicrobial activity was evaluated based on the minimum inhibitory concentration. The cytotoxicity was analyzed based on the hemolysing potential of the samples. The thermal degradation profile was performed by two different methods. Gallic acid was identified as the main compound of the samples and showed the highest amount in the chloroform fraction. All samples were able to inhibit the growth of the microorganisms tested and showed no cytotoxicity. The ethanol extract absorbs less heat than the fractions. All samples exhibited exothermic peak consistent with degradation of gallic acid. Based on the results, the samples used are potential candidates for use in dental formulations for biofilm control.

Made available in DSpace on 2021-06-25T11:46:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-20 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Federal Goiano Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundacao de Amaparo a Pesquisa do Estado de Goias (FAPEG, Brazil) Artemisinin extracted from Artemisia annua L. plants has a range of properties that qualifies it to treat several diseases, such as malaria and cancer. However, it has short half-life, which requires making continuous use of it, which has motivated the association of artemisinin (ART) with polymeric nanoparticles to increase its therapeutic efficiency. However, the ecotoxicological safety of this association has been questioned, given the scarcity of studies in this area. Thus, in this work the toxicity of Poly (epsilon-Caprolactone) nanocapsules added with ART (ART-NANO) in zebrafish (Danio rerio), embryos and adults was studied. Different endpoints were analyzed in organisms exposed to ART-NANO, including those predictive of embryotoxicity and histopatoxicity. Embryotoxicity was analyzed based on Organization for Economic Co-operation and Development (OECD) test guideline (236) for fish embryo acute toxicity applied to zebrafish (Danio rerio) at 96 hpf under five nominal logarithmic concentrations (0.125 to 2.0 mg/L). Our results demonstrate, mainly, that fertilized eggs presented increased coagulation, lack of heart rate, vitelline sac displacement and lack of somite formation. On the other hand, adult individuals (exposed to the same concentrations and evaluated after 24 and 96 h of exposure) have shown increased pericarditis. Therefore, the treatment based on ART, poly (epsilon-caprolactone) nanocapsules and on their combination at different concentrations have shown toxic effects on zebrafish embryos and adult individuals. (C) 2020 Elsevier B.V. All rights reserved. Univ Sao Paulo, Biomed Sci Inst, Pharmacol Dept, Sao Paulo, SP, Brazil Fed Univ Rio Janeiro UFRJ, Biodivers & Sustainabil Inst NUPEM, Integrated Translat Biosci Lab LIBT, Macae, RJ, Brazil Univ Fed Sao Carlos, Dept Genet & Evolut, Sao Carlos, SP, Brazil Univ Estadual Campinas, Pharmaceut Sci Sch, Campinas, SP, Brazil Univ Fed Minas Gerais, Belo Horizonte, MG, Brazil Sao Paulo State Univ, Jaboticabal, SP, Brazil Univ Fed Alfenas, Motric Sci Inst, Alfenas, MG, Brazil Livestock & Supply, Minist Agr, Sao Carlos, SP, Brazil Sao Paulo State Univ, Botucatu, SP, Brazil Goiano Fed Inst, Biol Sci Dept, Urutai, Go, Brazil Sao Paulo State Univ, Jaboticabal, SP, Brazil Sao Paulo State Univ, Botucatu, SP, Brazil CNPq: 426531/2018-3 Instituto Federal Goiano: 23219.000865.2020-78 CNPq: 307743/2018-7

Made available in DSpace on 2021-06-25T11:04:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-01 The guided tissue regeneration (GTR) technique can be applied in dentistry and other medical specializations, such as orthopedics. In modern dentistry, GTR has been used in periodontics and implantology to treat periodontal defects, to reconstruct lost, damaged and atrophied bone tissue in dental implant procedures, and to preserve alveolar bases after tooth extraction. In order to create and improve new therapies and to develop new biomaterials that restore, improve and prevent aggravation of compromised tissue function, poly (ϵ-caprolactone) (PCL) polymer membranes were obtained by the electrospinning process and were associated with two plant extracts: Pterodon pubescens Benth (P. pubescens) and Arrabidaea chica Verlot (A. chica) which are characterized by their pharmacological activities of anti-inflammatory and healing actions, respectively. Fiber morphology was analyzed using scanning electron microscopy (SEM), where fiber average diameter was measured from SEM images. Contact angle measurements were performed in order to evaluate the hydrophilicity of electrospun membranes containing vegetal extract. High-performance liquid chromatography was used to evaluate the ability to release active ingredients. Cytotoxicity and cell proliferation assays were performed in vitro on NIH-3T3 cells for 1, 3 and 7 d. Electrospun PCL membranes associated with plant extracts P. pubescens and/or A. chica presented a controlled release profile of the active compounds induced fibroblast formation, suggesting that they are promising and suitable for applications in GTR. Department of Manufacturing and Materials Engineering School of Mechanical Engineering University of Campinas UNICAMP School of Pharmacy Queen's University Belfast Graduate School of Bioscience and Technology of Bioactive Products Biology Institute University at Campinas Faculdade de Ciências Médicas Universidade Estadual de Campinas UNICAMP Faculty of Pharmaceutical Science University at Campinas Instituto de Biociencias Univ Estadual Paulista UNESP Instituto de Biociencias Univ Estadual Paulista UNESP

This study evaluated water sorption, solubility and compound release of 2.5% Arrabidaea chica polyphenolic extract (Ac) incorporated into the primer of a total-etch adhesive system; and dentin metalloproteinases inhibition when using Ac as dentin pretreatment in aqueous solution or incorporated either into acid or adhesive system primer. Adhesive system discs (Adper Scotchbond Multipurpose/3 M Oral Care/SB) with and without Ac were prepared (n = 10) for water sorption, solubility and compound release. Concentration of extract released in water was evaluated at 1, 3, 7, 14, 28, 36 days immersion. Dentin-resin interfaces were prepared for zymography analysis (control (SB); dentin pretreatment with water; dentin pretreatment with 2.5% Ac aqueous solution; Ac incorporated into phosphoric acid; Ac incorporated into SB primer) and assessed for in situ zymography tests after 24-h and 6-month storage. Primer incorporated with Ac showed lower water sorption (p = 0.0102) and solubility (p = 0.0065) than non-incorporated. Ac release was observed at third and seventh days (p = 0.0031). There was no significant difference between treatments (p = 0.4298) either immediately or after 6 months regarding relative fluorescence unit means among groups, and these means decreased after 6 months (p

The present study evaluated whether a natural dietary additive, dried Artemisia annua leaves, may be useful to control Rhipicephalus (Boophilus) microplus on naturally infested cattle. Twenty heifers of the Canchim breed, weighing around 250 kg, were divided into two equally sized experimental groups: 1) control animals and 2) animals receiving 200 g/day of dried A. annua leaves for two months. Before treatment began, the animals were homogeneously distributed in control and treatment groups based on their pre-treatment weight and tick infestation level. Counts of engorged female ticks then occurred weekly during the two-month experimental period. We also monitored cattle weight gain and packed cell volume (PCV). Artemisinin (0.96%) was quantified in the plant material by high-performance liquid chromatography with refractive index detector (HPLC-IR). No statistical differences between the control and treatment groups were observed for engorged female counts (log averages of 1.3 ticks and 1.4 ticks per animal, respectively), daily cattle weight gain (0.910 kg and 0.888 kg, respectively) or PCV (33.5% and 33.0%, respectively). We conclude that the oral supplementation of cattle feed with dried A. annuna leaves did not control natural infestation of R. (B.) microplus. The hypothesis of artemisinin’s action on cattle ticks by ingestion through the animals’ blood was not confirmed at the evaluated dose.

Arrabidaea chica, popularly known as “crajiru”, has three major pigments of the 3-deoxyanthocyanin type. Four types of extracts and two conditions of fractionation were used in order to obtain carajurin-rich extract fractions. In the first and second cases, the extracts were obtained with supercritical carbon dioxide (scCO2) from leaves and, from a dry hydroalcoholic extract (SDHAE), in the third the extract was obtained from the ethanolic extraction of a SDHAE, and in the fourth a reconstituted dry hydroalcoholic extract was used. Extract fractionation was carried out by stepwise pressure reduction in four separators in series, and in the third and fourth cases scCO2 was added in the first separator at a CO2/extract ratio of 91:9 (w/w). From the extraction of the leaves, extract fractions containing concentrated carajurin were obtained. From the ethanolic extraction of the SDHAE obtained some fractions concentrated in anthocyanins and fractionation of the reconstituted hydroalcoholic extract was inefficient.

Pterodon pubescens Benth is a Brazilian native species, used in folk medicine for chronic pain relief. The anti-inflammatory, antinociceptive, and antiproliferative activities attributed to the species have a straight relationship with vouacapans (furanditerpenes) and geranylgeraniol (acyclic diterpenes), identified in P. pubescens extracts. Nevertheless, scarce reports are found on product development and stability studies containing this species. This gap prompted the stability outcome evaluation of carbomer-940 gel-based formulations containing P. pubescens microencapsulated extract compared to non-encapsulated extract reported herein. Seeds were extracted with dichloromethane providing 46.6% (w/w) crude extract yield. Phytochemical markers were monitored by High Pressure Liquid Chromatography with Diode Array Detector (HPLC-DAD) and Gas Chromatography coupled to Mass Detection (GC-MS) using the vouacapans 1:1 ratio isomers 6α-hydroxy-7β-acetoxy-vouacapan-17β-oate methyl ester and 6α-acetoxy-7β-hydroxy-vouacapan-17β-oate methyl ester for quantification and identification purposes. Spray-dried microcapsules were produced with cashew gum as wall material with structure confirmed by scanning electron microscopy (SEM) and size distribution by light scattering parameters. The particles presented a bimodal size distribution ranging from 0.2 to 500 μm, vouacapans content and encapsulation efficiency of 55.6% and 44.4% respectively, with an overall spray-drying yield of 82.6% (w/w). The formulation containing microencapsulated extract did not achieve an acceptable outcome under test condition, with further studies needed for optimization. Nevertheless, the gel containing 2.5% (w/v) of non-encapsulated extract stored at 25 °C protected from light presented the best results within 180 days in stability study. P. pubescens incorporation maintained rheological properties with better performance when exposed to light during storage time compared to the gel base formulation. The formulation delivered vouacapans throughout a synthetic membrane. This formulation is the first step to achieve a novel topical gel-based product containing P. pubescens beneficial activities.

Herein, the antiproliferative potential of the essential oil obtained from fresh leaves of Croton campestris against human tumour cell lines was investigated for the first time. Furthermore, the essential oil obtained by hydrodistillation had the composition determined by gas chromatography–mass spectrometry (GC/MS). Ten major components were identified that comprised 91.59% of the total content, with 23.8% consisting of (Z)-caryophyllene and 16.08% consisting of γ-elemene as main components. The cytotoxic activity was observed mainly for breast (MCF-7) and colon (HT-29) human tumour cell lines, with GI50 (50% growth inhibition) concentration of 8.61 and 9.94 μg/mL, respectively. The results of this study showed that the essential oil obtained from Croton campestris A.St.-Hil. represents a potential source for the search of new antitumour agents.

Baccharis dracunculifolia is a plant native to Brazil, rich in phenolic compounds such as 3,5-diprenyl-4-hydroxycinnamic acid (artepillin C), a derivative of cinnamic acid, and 4-methoxy-3,5,7-trihydroxyflavone (kaempferide), a flavonoid. This study evaluated the performance of supercritical carbon dioxide (scCO 2 ) as solvent for selective fractionation by precipitation from a supercritical extract of B. dracunculifolia using scCO 2 pressure reduction in a sequence of separators to obtain different extract fractions in relation to its composition. The extracts and fractions were analyzed in terms of overall yield, antioxidant activity and concentration of protocatechuic acid, caffeic acid, sinapic acid, chrysin, p-coumaric acid, kaempferide and artepillin C. High concentrations were observed of the two major compounds (kaempferide and artepillin C) in the extract fractions of the first two separators. However, the aqueous extract obtained in the third step presented better antioxidant activity when compared the other fractions. Also obtained was a volatile fraction composed mainly of sesquiterpenes.

Artemisia annua L. is a well-known source of artemisinin, an antimalarial drug. This compound has been used in traditional medicine to treat malaria for many years. The anthelmintic property of A. annua has also been attributed to artemisinin, hence the belief that artemisinin could be useful as an alternative anthelmintic to control Haemonchus contortus in livestock. The present study focused on evaluating the anthelmintic activity of A. annua against H. contortus by oral administration on infected sheep in a controlled clinical trial. The trial was conducted using 24 male sheep of the Santa Ines breed, with average weight of 20 kg, artificially infected with H. contortus (Embrapa 2010 isolate). The animals were kept in metabolic cages and randomly divided into four homogeneous groups with six animals each, being: (T1) negative control (untreated), (T2) positive control: 10 mg/kg BW of levamisole phosphate, (T3) fed 10% A. annua (0.2% BW), (T4) fed 20% A. annua (0.4% BW). EPG counts were conducted on days -3, -2, -1, 0 (treatment day), 3, 7, 10, 14, 17, 21, 24 and 28 post-treatment. After extraction of the artemisinin in plant material, it was quantified in triplicate and analyzed by high-performance liquid chromatography with infrared detection. The average efficacy in the positive control treated with levamisole was 89%. Moderate anthelmintic efficacy against H. contortus was observed in the groups fed 0.2 and 0.4% BW A. annua for 30 days (32.9 and 14.8%, respectively), mainly from day 21 post treatment (47.1 and 25.2%, respectively). At the highest dose, the animals avoided eating due to the compound’s bitter taste. It is concluded that A. annua presents moderate anthelmintic activity in sheep in both doses. The lowest dose was accepted by animals and seems to have practical use. Oral administration was safe and after further in vivo trials could be introduced in organic farms in tropical countries. Key words: Artemisinin, sesquiterpenoids, gastrointestinal nematodes, control, veterinary ethnopharmacology.

This study evaluated the phytochemical characterization of Bixa orellana (BO extract) unsaponifiable extract and resulting fractions (F fraction - FF, Geranyl fraction – GF and R fraction- RF) obtained as by-products of an industrial process investigating in vitro antiproliferative activities in human tumoral cells. The main compounds identified by GC-MS for BO extract were Geranylgeraniol (61.51%); for FF: Geranylgeraniol (70.23%); for GF: Geranylgeraniol (78.92%) and for RF: β-cubebene (27.75%). Quantifications of geranylgeraniol by GC-FID presented the percentage content: BO 27.52%; FF 38.52%; GF 51.44% and RF 1.81%. BO extract showed a significant antiproliferative activity, with GI50 up to 4 µg/mL. All fractions had a remarkably similar antiproliferative activity profile (GI50 27-47 µg/mL). Data reported herein showed an important cytostatic effect for BO extract, nevertheless this activity is not attributed exclusively to geranylgeraniol. In conclusion, this by-product becomes of great value, being a potential candidate for development of new anti-tumor ingredients.

The combination of chitosan (C) with alginate (A) has been explored for the production of dressings due to the positive results on wound healing. CA films can show a dense or porous flexible structure, with characteristics tunable for different applications. Porosity and flexibility can be achieved, respectively, by the addition of surfactants such as Kolliphor® P188 (P) and silicone-based compounds as Silpuran® 2130 A/B (S). Furthermore, composite matrices of these polysaccharides have potential applications as devices for releasing bioactive compounds to skin lesions. The purpose of this study was to evaluate the physicochemical and biological characteristics of flexible dense and porous CA membranes incorporating the standardized extract of Arrabidaea chica Verlot (A. chica), and also to analyze the release mechanism of the extract from different membrane formulations. The results show that the inclusion of P in the formulation allows obtaining porous matrices, promotes greater homogeneity of the mixture of the silicone gel with the suspension of polysaccharides, and increases the swelling of the polymer matrix. All formulations presented high stability, reaching a maximum mass loss of 18% after seven days. The formulations with S showed the best performance in terms of flexibility and strain at break. The presence of A. chica standardized extract did not affect negatively the characteristics of the membranes. Incorporation efficiencies of the bioactive compound above 87% were achieved, and the addition of P and S to the membrane formulation changed the release of the A. chica extract kinetics. In addition, the developed formulations did not significantly affect Vero cells proliferation.

The aim of this study was to develop a phytocosmetic sunscreen emulsion with antioxidant effect, containing a blend of flavonoid-enriched plant extracts. In vitro sun protection factor, antioxidant activity, skin irritation, photostability, cutaneous permeation, and retention of flavonoids were evaluated. Thermodynamically stable emulsions were obtained and tested for sensorial analysis after loading the blend of extracts. The selected emulsion was stable when stored at low temperatures (5 C), for which after 120 days the concentration of quercetin and rutin were above their limit of quantification, i.e., 2.8 &plusmn, 0.39 &micro, g/mL and 30.39 &plusmn, g/mL, respectively. Spreadability, low rupture strength and adhesiveness were shown to be similar to a conventional topical product. Higher brittleness, pseudo-plastic, and viscoelastic behaviors were also recorded for the developed phytocosmetic sunscreen. The product presented a critical wavelength of 387.0 nm and ultraviolet rays A and B (UVA/UVB) rate of 0.78, confirming that the developed formulation shows capacity for UVA/UVB protection, protecting skin against damages caused by Ultraviolet (UV) radiation. Rutin was shown to permeate the skin barrier and was also quantified in the stratum corneum (3.27 &plusmn, 1.92 &micro, g/mL) by tape stripping and retention test (114.68 &plusmn, 8.70 &micro, g/mL). The developed flavonoid-enriched phytocosmetic was shown to be non-irritant to skin by an in vitro assay. Our results confirm the antioxidant activity, sun protection, and physical properties of the developed phytocosmetic for topical application.

Made available in DSpace on 2020-12-12T02:44:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The aim of this paper was to evaluate the physical properties and the long-term bond strength of a 2.5% polyphenol-enriched extract of Arrabidaea chica (AC) incorporated into both the phosphoric acid and the primer of a three-step total-etch adhesive, or into an aqueous solution as a dentin pretreatment. Fifty dentin surfaces received the treatments (n = 10): CON (control) – application of the three-step adhesive system (Adper Scotchbond Multipurpose, 3M ESPE); WAT – distilled water used as a pretreatment after dentin etching and before application of the adhesive system; ACPA - AC incorporated into the phosphoric acid; ACW - dentin pre-treatment with AC incorporated into an aqueous solution after etching; ACP - AC incorporated into the primer. Microtensile bond strength tests were performed after 24 h, 6 and 12 months of storage. Slices from the resin-dentin interface were obtained for scanning electron microscopy analysis of the hybrid layer. Degree of conversion of AC incorporated into the primer was evaluated. The particle size, polydispersity index and zeta potential of all the solutions prepared by incorporating AC (phosphoric acid, primer and distilled water) were measured by dynamic light scattering, which brought about changes after incorporation. Degree of conversion of the primer was not affected after incorporating AC. ACP showed lower microtensile bond strength values than the other groups. Bond strength decreased after 6 months of storage, stabilizing at the 12-month evaluation. Therefore, use of AC incorporated into the primer led to lower bond strength values, since AC modified the physical properties (particle size, polydispersity index and zeta potential) of the primer, but did not change the degree of conversion. Application of AC as a dentin pretreatment did not affect bond strength or the micromorphological characteristics of the hybrid layer. São Leopoldo Mandic Research Institute, Rua José Rocha Junqueira 13, Bairro Swift Faculty of Pharmaceutical Sciences University of Campinas, P.O. Box 6029 Institute of Science and Technology of São José dos Campos UNESP-University of the State of São Paulo, Avenida Engenheiro Francisco José Longo, 777, Jardim São Dimas Institute of Science and Technology of São José dos Campos UNESP-University of the State of São Paulo, Avenida Engenheiro Francisco José Longo, 777, Jardim São Dimas FAPESP: 2016/04343-8 FAPESP: 2017/08082-7 CNPq: 303367/21018-0

This study evaluated the pharmacological effect of the association of crude extract from the fruits of Pterodon pubescens (Pp) with the essential oil of Cordia verbenacea (Cv) in antinociception and anti-inflammatory experimental models. The effective doses of each extract and the combinations used in the associations of extracts were defined by acetic acid-induced writhing test. The separate extracts were also evaluated on formalin test. Interaction between extracts was assessed by isobologram method. The effects of different concentrations of associations (A50, A100 and A200) were evaluated on formalin test, tail flick and hot plate. The anti-inflammatory activity was evaluated in paw edema induced by carrageenan and PGE2, carrageenan-induced peritonitis and mechanical allodynia induced by Complete Freund's Adjuvant (CFA). The associations were markedly synergistic, as assessed using isobolographic analyses. On formalin and on acetic acid-induced writhing tests, associations demonstrated greater efficacy when compared to extracts separately. In paw edema models, significant reductions of edema were observed. On mechanical allodynia induced by CFA, associations were effective at acute phase with pronounced effect at chronic phase. The associations were not effective in hot plate, tail flick and carrageenan-induced peritonitis tests. Phytochemical analysis by HPLC-DAD and FID showed important concentrations of α-Humulene, trans-Caryophyllene, geranylgeraniol, isomers 6α-hydroxy-7β-acetoxy-vouacapan-17βoate methyl ester and 6α-acetoxy-7β-hydroxy-vouacapan-17β-oate methyl ester (compounds m/z 404) and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester (m/z 362). These findings demonstrate that the associations promote synergistic antinociceptive effect and important anti-inflammatory activities, especially on chronic inflammation conditions, at lower doses than the separate crude extracts, without demonstrating side effects, probably acting in different pharmacological receptors. The inhibition of inflammation suggests a relationship with inflammatory mediators and PGE2 pathway and might be exploited to achieve greater anti-inflammatory efficacy, being considered as a potential phytotherapy.

Produção de extratos vegetais padronizados e partículas contendo extrato encapsulada em micro e/ou nanopartículas, preparo de formulações com avaliação dos parâmetros de estabilidade e estudo da capacidade cicatrizante.

Solubility data of some phenolic compounds of industrial interest, such as protocatechuic acid (3,4-dihydroxybenzoic acid), chrysin (5,7-dihydroxyflavone) and sinapic acid (3,5-dimethoxy-4-hydroxycinnamic acid) in supercritical carbon dioxide were determined at pressures of 200, 300 and 400 bar and temperatures of 40, 50 and 60 °C. Experimental data were correlated with semi-empirical Chrastil model and thermodynamic modeling using the Peng–Robinson equation of state with classical mixing rule. The critical properties, acentric factor, and vapor pressure of these compounds were estimated by group contribution methods. The solubility values followed a similar tendency, with higher solubility with increasing temperature and pressure, and no crossover pressure in the range studied. However, a crossover pressure around 150 bar was observed when extrapolating the solubility data by the Chrastil equation, which is in agreement with values reported in the literature for phenolic compounds, thereby indicating the consistency of the solubility experimental data.

Varronia curassavica Jacq. is a native medicinal plant from Brazil, its anti-inflammatory, analgesic and healing properties are mainly due to the presence of α-humulene and β-cariophyllene on the essential oil extracted from the leaves. The study aimed to evaluate parameters related to the production, harvest and extraction and their effects on the quality/quantity of the V. curassavica essential oil under field conditions. For the drying experiment it was used natural drying in greenhouse and an artificial one with forced ventilation using the temperature of 40 °C until constant weight with six replications per treatment. For the harvest daytime experiment, five harvest different times were evaluated (6 a.m., 9 a.m., 12 p.m., 3 p.m., 6 p.m.) with five replications per treatment. In both experiments it was used the completely randomized design. The effect of two natural elicitors on the sesquiterpene content of V. curassavica was also evaluated, where field grown plants received the application of acibenzolar-S-methyl (500 mg/L), 1,6 β- d -glucan obtained from Lasiodiplodia theobromae (50 mg/L) and distilled water as a control, where gas exchange rate, terpene enzymes such as phenylalanine ammonia-lyase, superoxide dismutase, guaiacol peroxidase and catalase activity and essential oil content in leaves were measured. The exhaustion time for the extraction of the V. curassavica essential oil by hydrodistillation was determined for whole and ground leaves. To evaluate and compare the extraction techniques performance, it was used a conventional and a green technology, it was evaluated the essential oil yield and α-humulene content in the volatile oil. Finally, a study of economic viability of the implantation of an agroindustry to obtain V. curassavica essential oil was performed using Expanded Multi-index Methodology (EMIM). The essential oil yielding was determined through hydrodistillation, using a Clevenger device, and the essential oil quality was measure by Gas Chromatography. There was not statistical difference between the drying methods tested regarding the essential oil production but the highest values of α-humulene and β-caryophyllene were obtained when artificial drying at 40 °C was adopted. It is possible to indicate a harvesting daytime around 11–12 a.m., resulting not only in higher yield of essential oil but also a higher content of the desired anti-inflammatory compounds. The acibenzolar-S-methyl reduced significantly the net carbon assimilation rate and the intercellular CO2 concentration, while1,6 β- d -glucan reduced significantly only the intercellular CO2 concentration. The essential oil analysis reveals that elicitation was not efficient to increase the yield of V. curassavica essential oil or to change relative rate among the oil chemical constituents, especially α-humulene and β-caryophyllene. The determined exhaustion time was 1.5 h for both treatments, and the conventional extraction technology (steam distillation) is the best option for extraction of the essential oil. At last, according to the economic study, it is recommended the implementation of the investment project.

In the context of the cancer-inflammation relationship and the use of natural products as potential antitumor and anti-inflammatory agents, the alkaloid-enriched fraction of Boehmeriacaudata (BcAEF) aerial parts was evaluated. In vitro antiproliferative studies with human tumor cell lines showed high activity at low concentrations. Further investigation on NCI-H460 cells showed an irreversible effect on cell proliferation, with cell cycle arrest at G2/M phase and programmed cell death induction. Molecular docking studies of four alkaloids identified in BcAEF with colchicine&rsquo, s binding site on &beta, tubulin were performed, suggesting (&minus, )-C (15R)-hydroxycryptopleurine as the main inductor of the observed mitotic death. In vivo studies showed that BcAEF was able to reduce Ehrlich tumor volume progression by 30 to 40%. Checking myeloperoxidase activity, BcAEF reduced neutrophils migration towards the tumor. The in vivo anti-inflammatory activity was evaluated by chemically induced edema models. In croton oil-induced ear edema and carrageenan (CG)-induced paw edema models, BcAEF reduced edema around 70 to 80% together with inhibition of activation and/or migration of neutrophils to the inflammatory area. All together the results presented herein show BcAEF as a potent antitumor agent combining antiproliferative and anti-inflammatory properties, which could be further explored in (pre)clinical studies.

The aim of this study was to validate a HPLC method for the assay of flavonoids in extracts obtained from natural sources, i.e., from Dirmophandra mollis Benth, Ginkgo biloba L., Ruta graveolens L., and Vitis vin&iacute, fera L. The potential sun protecting effect, antioxidant activity, and cell viability of the extracts were also determined. Individual extracts (obtained from each individual species) and a mixed extract (containing the four extracts) were analyzed by the validated HPLC method for the identification of flavonoids and quantification of rutin and quercetin. An in vitro cell viability study was carried out using the neutral red method. The in vitro sun protection factor was determined by spectral transmittance and in vitro antioxidant efficacy was evaluated against DPPH, ABTS, and AAPH radicals. The HPLC method used for the identification and quantification of flavonoids in extracts exhibited linearity, precision, accuracy, and robustness. Detection and quantification limits were, respectively, 2.881 &plusmn, 0.9 &mu, g&middot, mL&minus, 1 and 0.864 &plusmn, 1 for quercetin, and 30.09 &plusmn, 1 &mu, 1 and 9.027 &plusmn, 1.1 &mu, 1 for rutin. All extracts did not affect cell viability at the evaluated concentration range and exhibited a sun protection effect and antioxidant activity. Among the evaluated extracts, Ginkgo biloba L. and the mixed extract depicted the most expressive antioxidant activity. The mixed extract exhibited sunscreen protection against ultraviolet A (UVA) and ultraviolet B (UVB) and a critical wavelength of 372.7 &plusmn, 0.1. Our results translate the enhanced flavonoids&rsquo, composition of the mixed extract, which may be a potential alternative over sunscreens and antioxidants in pharmaceutic/cosmetic formulations.

To evaluate the effects of the bed geometry on the scale-up of fixed bed extractors and on the cost of supercritical extracts from Baccharis dracunculifolia leaves (local name in Brazil: alecrim-do-campo), supercritical CO2 (scCO2) extraction curves at 50 °C and 300 bar at different heights (H) and bed diameter (D), at H/D ratio ranging from 1.86 to 5.2 were obtained. The global yield in all experiments was around 4.6%, and the scale-up criterion adopted was maintain constant the solvent flux. Five phenolic compounds of interest, caffeic acid, p-coumaric acid, trans-cinnamic acid, artepillin C, and kaempferide were quantified by high-performance liquid chromatography. The fractions exhibited high kaempferide levels, which varied from 29.07 to 46.47 mg kaempferide per gram of extract. The optimal manufacturing cost has been estimated at US $ 171.12/kg at 80 min, which shows the possibility of obtaining B. dracunculifolia extracts at an industrial level.

The aim of this study was to evaluate in vitro the antifungal, antibiofilm and antiproliferative activities of the extract from the leaves of Guapira graciliflora Mart. The phytochemical characterization of the extract was performed using electrospray ionization mass spectrometry (ESI-MS). The antimicrobial activity of the extract and its fractions was evaluated using the broth microdilution method against species of Candida. The inhibition of C. albicans biofilm was evaluated based on the number of colony-forming units (CFU) and metabolic activity (MTT). The antiproliferative activity of the extract and its fraction was evaluated in the presence of human tumor and non-tumor cells, and the cytotoxicity of the extract was determined on the RAW 264.7 macrophage line – both using the sulforhodamine B method. The phytochemical characterization indicated the presence of the flavonoids rutin and kaempferol. The extract and the methanol fraction exhibited moderate antifungal activity against C. albicans, C. krusei, and C. glabrata, and strong activity against C. dubliniensis. In the biofilms at 24 and 48 hours, the concentration of 12500 µg/mL of the extract was the most effective at reducing the number of CFU s/mL (44.4% and 42.9%, respectively) and the metabolic activity of C. albicans cells (34.6% and 52%, respectively). The extract and its fractions had no antiproliferative effect on the tumor lines tested, with mean activity (log GI50) equal to or greater than 1.71 µg/mL. Macrophage cell viability remained higher than 80% for concentrations of the extract of up to 62.5 µg/mL. G. graciliflora has flavonoids in its chemical composition and demonstrates potential antifungal and antibiofilm activity, with no evidence of a significant change in the viability of human tumor and non-tumor cell lines.

Pterodon pubescens Benth., popularly known as "sucupira", is traditionally used as pain healing agent for many inflammatory diseases. The present study evaluated the in vivo antinociceptive and anti-inflammatory properties of sucupira's fruit dicloromethane extract (Pp) compared to the aqueous extract (Ppa) traditionally used in folk medicine. Extracts' chemical characterization was performed by gas chromatography coupled to mass spectra (GC-MS) detection. The standardized extracts were evaluated using antinociceptive and anti-inflammatory experimental models with mice. The results reported herein allowed establishing a relationship between the popular use of Pterodon pubescens fruit for pain relief and the activity of two major compounds isolated from this species which demonstrated antinociceptive activity. The experimental models corroborate activity of aqueous extract antinociceptive and anti-inflammatory activity, with lower potency compared to dichloromethane extract. Nevertheless the resulting data corroborates sucupira's folk use for pain relief.

There is no effective natural alternative control for gastrointestinal nematodes (GIN) of small ruminants, with Haemonchus contortus being the most economically important GIN. Despite frequent reports of multidrug-resistant GIN, there is no new commercial anthelmintic to substitute failing ones. Although trematocidal activity of artemisinin analogs has been reported in sheep, neither artemisinin nor its plant source (Artemisia annua) has been evaluated for anthelmintic activity in ruminants. This study evaluated the anthelmintic activity of A. annua crude extracts in vitro and compared the most effective extract with artemisinin in sheep naturally infected with H. contortus. A. annua leaves extracted with water, aqueous 0.1% sodium bicarbonate, dichloromethane, and ethanol were evaluated in vitro by the egg hatch test (EHT) and with the bicarbonate extract only for the larval development test (LDT) using H. contortus. The A. annua water, sodium bicarbonate (SBE), ethanol, and dichloromethane extracts tested in vitro contained 0.3, 0.6, 4.4, and 9.8% of artemisinin, respectively. The sodium bicarbonate extract resulted in the lowest LC99 in the EHT (1.27 μg/mL) and in a LC99 of 23.8 μg/mL in the LDT. Following in vitro results, the SBE (2 g/kg body weight (BW)) and artemisinin (100 mg/kg BW) were evaluated as a single oral dose in naturally infected Santa Inês sheep. Speciation from stool cultures established that 84-91% of GIN were H. contortus, 8.4-15.6 % were Trichostrongylus sp., and 0.3-0.7% were Oesophagostomum sp. Packed-cell volume and eggs per gram (EPG) of feces were used to test treatment efficacy. The SBE tested in vivo contained no artemisinin, but had a high antioxidant capacity of 2,295 μmol of Trolox equivalents/g. Sheep dosed with artemisinin had maximum feces concentrations 24 h after treatment (126.5 μg/g artemisinin), which sharply decreased at 36 h. By day 15, only levamisole-treated sheep had a significant decrease of 97% in EPG. Artemisinin-treated and SBE-treated sheep had nonsignificant EPG reductions of 28 and 19%, respectively, while sheep in infected/untreated group had an average EPG increase of 95%. Sheep treated with artemisinin and A. annua SBE maintained blood hematocrits throughout the experiment, while untreated/infected controls had a significant reduction in hematocrit. This is the first time oral dose of artemisinin and an aqueous extract of A. annua are evaluated as anthelmintic in sheep. Although oral dose of artemisinin and SBE, at single doses, were ineffective natural anthelmintics, artemisinin analogs with better bioavailability than artemisinin should be tested in vivo, through different routes and in multiple doses. The maintenance of hematocrit provided by artemisinin and A. annua extract and the high antioxidant capacity of the latter suggest that they could be combined with commercial anthelmintics to improve the well-being of infected animals and to evaluate potential synergism.

Phenolic compounds from Arrabidaea chica Verlot leaves, besides conferring staining properties to their extracts, also have various biological activities including anti-inflammatory and wound-healing properties. To evaluate new possibilities for obtaining extracts with differentiated yield and composition, sequential extractions in fixed bed were performed at 40 and 50 °C, and 300 and 400 bar, using as extracting solvents pure supercritical carbon dioxide (scCO 2 ) in a first step, acidified ethanol in a second step and acidified water in a third extraction step. Four flavonoids of interest were investigated in the extracts, one of them being flavone (luteolin), and three anthocyanin compounds of type 3-desoxyanthocyanidins which were quantified by high performance liquid chromatography (HPLC). The extraction curves, the global yields and the concentration and yield of the compounds under study were evaluated. The results indicated that the cumulative total yields in the three steps ranged from 22% to 27% in all conditions of temperature and pressure, with the highest global yield at 50 °C and 300 bar. Although the lowest extraction yield was obtained using pure scCO 2 , this step was highly selective, since only carajurin in its aglycone form was extracted among the compounds of interest and this was confirmed by analysis of MS/MS.

O presente estudo objetivou estimar a variabilidade genética existente entre caracteres agronômicos, fisiológicos e fitoquímicos em variedades de A. annua. O delineamento experimental foi inteiramente casualizado e os tratamentos foram as variedades Artemis, 2/39x5x3M, e 2/39x1V de A. annua, submetidas a avaliações agronômicas, fisiológicas e fitoquímicas. Para a realização das estimativas de distância genética foram geradas matrizes de dissimilaridade utilizando a distância Euclidiana e os métodos de agrupamento de Tocher e UPGMA. Além disso, avaliou-se a importância relativa dos caracteres para divergência genética pelo método de Singh. As análises foram realizadas pelo software Genes e os dendrogramas obtidos pelo NTSYS. A presença de variabilidade genética dentro das variedades permitiu a identificação de acessos dissimilares e com média elevada para as características estudadas. O número de ramificações, concentração intracelular de CO2, e o rendimento de óleo essencial foram os caracteres que mais contribuíram para a dissimilaridade genética de A. annua. Os acessos B24, C5 e C32 foram os mais promissores dentro das variedades e devem ser conservados para futuras hibridações, sendo que as hibridações mais promissoras na obtenção de populações segregantes desejadas são B24 x C5, B24 x C32 e C5 x C32. This study aimed to estimate the genetic variability among agronomic, physiological, and phytochemical characters in varieties of A. annua. The experimental design was completely randomized and the treatments were the varieties Artemis, 2/39x5x3M and 2/39x1V of A. annua, subject to agronomic, physiological and phytochemical evaluations. To estimate the genetic distances, dissimilarity matrices were generated using the Euclidean distance and the Tocher and UPGMA grouping methods. Moreover, we evaluated the relative importance of the characters for genetic divergence through the method of Singh. The analyses were performed in the Genes software and the dendrograms were obtained from the NTSYS program. The presence of genetic variability within the varieties allowed the identification of dissimilar accessions with high average for all traits. The number of branches, intracellular concentration of CO2 and oil yield were the traits that contributed most to the genetic dissimilarity of A. annua. The accessions B24, C5 and C32 were the most promising within the populations and must be conserved for future crossings, and the most promising crosses to obtain the desired segregant populations were B24 x C5, B24 x C32 and C5 x C32.