From Leaves to Reproductive Organs: Chemodiversity and Chemophenetics of Essential Oils as Important Tools to Evaluate Piper mollicomum Kunth Chemical Ecology Relevance in the Neotropics.

Piper mollicomum Kunth (Piperaceae) plays a vital role in the preservation of the Brazilian Atlantic Forest by contributing to the regeneration of deforested areas. Recent scientific investigations have analyzed the chemical constituents and seasonal dynamics of essential oils (EO) from various Piper L. species, highlighting the need to elucidate their chemical-ecological interactions. This study aims to expand the chemical-ecological knowledge of this important taxon in neotropical forests, using P. mollicomum as a model. The methodologies employed include the collection of plant material, EO extraction by hydrodistillation, analysis of EO by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detector (GC-FID), recording the frequency of visits by potential pollinators and microclimatic variables, and by conducting calculations of chemodiversity and chemophenetic indices. Chemical analyses indicated that the diversity of EO and environmental factors are linked to the activities of potential pollinators. In the Tijuca Forest, P. mollicomum revealed significant interactions between its volatile constituents and microclimatic variables, showing that the chemodiversity of the leaves and reproductive organs correlates with pollinator visitation. Additionally, a notable difference in chemical evenness was observed between these vegetative structures. The chemophenetic indices by Ramos and Moreira also revealed correlations with chemical diversity.

Spatio-Temporal Variations of Volatile Metabolites as an Eco-Physiological Response of a Native Species in the Tropical Forest.

This study evaluates the essential oil (EO) composition of Piper rivinoides Kunth, a shrub native to the Brazilian tropical rainforest, across different plant parts and developmental phases. The aim was to explore the chemical diversity of EO and its reflection in the plant's ecological interactions and adaptations. Plant organs (roots, stems, branches, and leaves) at different developmental phases were subjected to hydrodistillation followed by chemical analysis using Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography-Flame Ionization Detector (GC-FID). The results revealed a relevant variation in EO yield and composition among different plant parts and developmental phases. Leaves showed the highest yield and chemical diversity, with α-pinene and ß-pinene as major constituents, while roots and stems were characterized by a predominance of arylpropanoids, particularly apiol. The chemical diversity in leaves increased with plant maturity, indicating a dynamic adaptation to environmental interactions. The study underscores the importance of considering the ontogeny of plant parts in understanding the ecological roles and potential applications of P. rivinoides in medicine and agriculture. The findings contribute to the overall knowledge of Piperaceae chemodiversity and ecological adaptations, offering insights into the plant's interaction with its environment and its potential uses based on chemical composition.

Insecticidal activity of essential oils from Piper aduncum against Ctenocephalides felis felis: a promising approach for flea control.

Piper aduncum L., a Brazilian medicinal plant, is known for its bioactive properties, including repellent and insecticidal effects. This study investigated the insecticidal potential of essential oils (EOs) from P. aduncum, collected during the dry and rainy seasons, against fleas (Ctenocephalides felis felis Bouché, 1835) in egg and adult stages. The EOs were obtained by hydrodistillation using a modified Clevenger apparatus for 2 h. Qualitative and quantitative analysis were performed via gas chromatography. The findings revealed that dillapiole was the predominant substance in both EOs, accounting for 77.6% (rainy) and 85.5% (dry) of the EOs. These EOs exhibited high efficacy against the parasite C. felis felis, resulting in 100% egg mortality at a concentration of 100 µg/mL and 100% mortality for adult fleas starting from 1,000 µg/mL. Dillapiole standard was also effective but at a relatively high concentration. This finding suggested that EOs from P. aduncum exhibit cytotoxicity against these pests and might hold potential for commercial production, offering practical applications for such bioprospecting. This study uniquely revealed that the EOs from P. aduncum, which is rich in dillapiole, demonstrated pulicidal activity against the parasite C. felis felis, particularly in inhibiting the hatching of the eggs of these parasites.

UHPLC-HRMS/MS Chemical Fingerprinting of the Bioactive Partition from Cultivated Piper aduncum L.

Piper aduncum L. is widely distributed in tropical regions and the ethnobotanical uses of this species encompass medicinal applications for the treatment of respiratory, antimicrobial, and gynecological diseases. Chemical studies reveal a diverse array of secondary metabolites, including terpenes, flavonoids, and prenylated compounds. Extracts from P. aduncum have shown antibacterial, antifungal, and larvicidal activities. Our study explores the activity of extracts and partitions against Mycobacterium tuberculosis H37Rv, as well as the chemical diversity of the bioactive partition. This marks the first investigation of the bioactive partition of P. aduncum from agroecological cultivation. The ethyl acetate partition from the ethanolic leaf extract (PAEPL) was found to be the most active. PAEPL was subjected to column chromatography using Sephadex LH-20 and the obtained fractions were analyzed using UHPLC-HRMS/MS. The MS/MS data from the fractions were submitted to the online GNPS platform for the generation of the molecular network, which displayed 1714 nodes and 167 clusters. Compounds were identified via manual inspection and different libraries, allowing the annotation of 83 compounds, including flavonoids, benzoic acid derivatives, glycosides, free fatty acids, and glycerol-esterified fatty acids. This study provides the first chemical fingerprint of an antimycobacterial sample from P. aduncum cultivated in an agroecological system.

Toxicological effects of aqueous extract of Genipa americana L. leaves on adult zebrafish (Danio rerio): Chemical profile, histopathological effects and lack of genotoxicity.

Genipa americana is a native plant of Brazil with potential applications in folk medicine. Whereas most of the phytochemical and pharmacological studies on this plant have focused on its fruits, the crude extracts of its leaves contain chemical metabolites that may have toxicity to organisms, which have yet to be investigated. This study aimed to determine the main groups of secondary metabolites in the aqueous extract of the leaves of G. americana by phytochemistry and qualitative HPLC, and to evaluate the possible toxicological effects and histopathological changes caused by this extract in zebrafish (Danio rerio) adults, through micronucleus test, nuclear abnormalities and histopathological analyses of gills and liver. While three metabolites of high intensity (phenolic compounds, flavonoids and triterpenes) were found in the phytochemical evaluation, the HPLC showed results compatible with flavonoids and iridoids, all belonging to common classes for this species and the Rubiaceae family. The acute toxicity test did not induce mortality or genotoxicity in zebrafish, but after exposure for 96 h, it was possible to observe injuries to the fish gill tissue, such as lamellar fusion, vasodilation and telangiectasia; in the liver, necrosis was visualized at 40 mg/L, and at higher concentrations (80 and 100 mg/L) induced sinusoidal widening was identified. In conclusion, the results demonstrated the toxic potential of this plant for aquatic species.

Bioproducts from Passiflora cincinnata Seeds: The Brazilian Caatinga Passion Fruit.

The present work aimed to obtain bioproducts from Passiflora cincinnata seeds, the Brazilian Caatinga passion fruit, as well as to determine their physical, chemical and biological properties. The seeds were pressed in a continuous press to obtain the oil, which showed an oxidative stability of 5.37 h and a fatty profile rich in linoleic acid. The defatted seeds were evaluated for the recovery of antioxidant compounds by a central rotation experimental design, varying temperature (32-74 °C), ethanol (13-97%) and solid-liquid ratio (1:10-1:60 m/v). The best operational condition (74 °C, 58% ethanol, 1:48) yielded an extract composed mainly of lignans, which showed antioxidant capacity and antimicrobial activity against Gram-positive and Gram-negative bacteria. The microencapsulation of linoleic acid-rich oil through spray drying has proven to be an effective method for protecting the oil. Furthermore, the addition of the antioxidant extract to the formulation increased the oxidative stability of the product to 30% (6.97 h), compared to microencapsulated oil without the addition of the antioxidant extract (5.27 h). The microparticles also exhibited favorable technological characteristics, such as low hygroscopicity and high water solubility. Thus, it was possible to obtain three bioproducts from the Brazilian Caatinga passion fruit seeds: the oil rich in linoleic acid (an essential fatty acid), antioxidant extract from the defatted seeds and the oil microparticles added from the antioxidant extract.

Development of a microemulsion loaded with epoxy-α-lapachone against Leishmania (Leishmania) amazonensis murine infection.

Epoxy-α-lapachone (ELAP), an oxirane-functionalized molecule synthesized from naturally occurring lapachol, has shown promising activity against murine infection with Leishmania (Leishmania) amazonensis. Herein, we report the successful development of oil-in-water-type (o/w) microemulsions (ME) loaded with ELAP (ELAP-ME) using Capmul MCM, Labrasol, and PEG 400. Stability studies revealed that ELAP-ME (100 µg/mL of ELAP), which was comprised of globule size smaller than 120.4 ± 7.7 nm, displayed a good stability profile over 73 days. ELAP-ME had an effect in BALB/c mice infected with L. (L.) amazonensis, causing reductions in paw lesions after two weeks of treatment (∼2-fold) when compared to untreated animals. Furthermore, there was also a reduction in the parasite load both in the footpad (60.3%) and in the lymph nodes (31.5%). Based on these findings, ELAP-ME emerges as a promising treatment for tegumentar leishmaniasis.

Is There Enough Knowledge to Standardize a Cannabis sativa L. Medicinal Oil Preparation with a High Content of Cannabinoids?

Introduction: Cannabis sativa L. medicinal oils are good therapeutic options due to their wide spectrum of pharmacological applications and the easy adjustment of individual doses. The lack of standardization of methodology in the preparation of medicinal oil using the Cannabis crude extract results in elevated variability of cannabinoid concentration in the final product. The elevated variability impairs the understanding of beneficial and adverse effects related to dose-response pharmacological activities. Objective: This study aimed to conduct a review on the current methods of Cannabis oil preparation present in the literature, to demonstrate the most appropriate methodologies to ensure a product with high content of cannabinoids and terpenes. Results: The decarboxylation stage is essential for the conversion of acid cannabinoids into neutral cannabinoids, which are substances with the highest bioavailability. Lower temperatures for longer periods of time instead of high temperatures in less time are highly recommended to ensure that all the acidic cannabinoids have passed through decarboxylation. For the guarantee of a high terpene content, the separate addition of essential oil to the fixed oil prepared from the crude extract should be considered. Ultrasound-assisted extraction is one of the best performing methodologies because it is cheaper than other techniques, such as supercritical fluid extraction, besides that, ultrasound extraction is effective in short extraction times and uses small amounts of solvent when compared with other techniques. Conclusion: Although the literature about the methods of preparation of Cannabis medicinal oil is scarce, it is possible to standardize an optimized, low-cost, and effective Cannabis extractive methodology from the results found in the literature; however, this will depend on new research for methodological validation.

Bauhinia forficata Link Infusions: Chemical and Bioactivity of Volatile and Non-Volatile Fractions.

This study aimed to evaluate Bauhinia forficata infusions prepared using samples available in Rio de Janeiro, Brazil. As such, infusions at 5% (w/v) of different brands and batches commercialized in the city (CS1, CS2, CS3, and CS4) and samples of plant material botanically identified (BS) were evaluated to determine their total phenolic and flavonoid contents (TPC and TFC), antioxidant capacity (ABTS•+, DPPH•, and FRAP assays), phytochemical profile, volatile compounds, and inhibitory effects against the α-amylase enzyme. The results showed that infusions prepared using BS samples had lower TPC, TFC and antioxidant potential than the commercial samples (p < 0.05). The batch averages presented high standard deviations mainly for the commercial samples, corroborating sample heterogeneity. Sample volatile fractions were mainly composed of terpenes (40 compounds identified). In the non-volatile fraction, 20 compounds were identified, with emphasis on the CS3 sample, which comprised most of the compounds, mainly flavonoid derivatives. PCA analysis demonstrated more chemical diversity in non-volatile than volatile compounds. The samples also inhibited the α-amylase enzyme (IC50 value: 0.235−0.801 mg RE/mL). Despite the differences observed in this work, B. forficata is recognized as a source of bioactive compounds that can increase the intake of antioxidant compounds by the population.

Umbu Fruit Peel as Source of Antioxidant, Antimicrobial and α-Amylase Inhibitor Compounds.

Herein, the extraction of bioactive compounds from umbu fruit peel was optimized using thermal-assisted solid-liquid extraction. In parallel, antioxidant, antimicrobial, and inhibitory effects against α-amylase of optimized extract were also evaluated. The combination of operational conditions including the temperature (32-74 °C), ethanol concentration (13-97%), and solid/liquid ratio (1:10-1:60; w/v) was employed using a rotational central composite design for optimization. The extracts were evaluated for total phenolic compounds (TPC), total flavonoid compounds (TFC) and antioxidant capacity by ABTS•+, DPPH• and FRAP assays. The bioactive profile of the optimized extract was obtained by ultra-performance liquid chromatography coupled to quadrupole/time-of-flight mass spectrometry in electrospray ionization in both negative and positive modes. The statistically evaluated results showed that the optimal operational conditions for the recovery of bioactive compounds from umbu fruit peel included 74 °C, 37% ethanol, and a solid-liquid ratio of 1:38. Under these conditions, the obtained values were 1985 mg GAE/100 g, 1364 mg RE/100 g, 122 µmol TE/g, 174 µmol/TE g and 468 µmol Fe2+/g for TPC, TFC, ABTS•+, DPPH•, and FRAP assays, respectively. In addition, the optimized extract was effective against Gram-positive and Gram-negative bacteria (MBC ranged from 0.060 to 0.24 mg GAE/mL), as well as it was effective to inhibit α-amylase (IC50 value of 0.076 mg GAE/mL). The optimized extract showed to be mainly constituted by phenolic acids and flavonoids.

Apoptotic effect of ß-pinene on oral squamous cell carcinoma as one of the major compounds from essential oil of medicinal plant Piper rivinoides Kunth.

Oral squamous cell carcinoma (OSCC) is the most common type of head and neck malignancy. Research on essential oils (EOs) has shown important cytotoxic and anti-tumor properties, among others. Piperaceae species are rich in EOs and here we highlight Piper rivinoides Kunth. We investigated the crude EOs from P. rivinoides, their pure major constituents and an enriched fraction with the main EO compounds (EF) as cytotoxic and selective OSCC agents. EOs presented as main compounds (-)-α-pinene, (-)-ß-pinene and limonene. EOs showed an IC50 lower than all isolated compounds, except for (-)-ß-pinene in OSCC cells. The (-)-ß-pinene induced cell death with apoptotic characteristics. Commercial standards showed greater selectivity than EOs, and (-)-ß-pinene was the most selective among them. EF showed higher selectivity compared to crude EOs and carboplatin, turning it into a good candidate as an anticancer fraction. These results are important for the possible development of new treatments for OSCC.

Advanced Chemophenetic Analysis of Essential Oil from Leaves of Piper gaudichaudianum Kunth (Piperaceae) Using a New Reduction-Oxidation Index to Explore Seasonal and Circadian Rhythms.

The aromatic species Piper gaudichaudianum Kunth (Piperaceae) is widely used in Brazil for medicinal and ritualistic applications. In the current study, chemophenetic patterns were realized across season and circadian rhythm based on the chemical profile of essential oils (EOs) from leaves. Hydrodistilled essential oils were analyzed by GC-MS and GC-FID, and a new calculation of metabolite oxidation level, averaged for each individual molecule component of the EO, was used to explore the patterns of metabolism/biosynthesis. This new index used an intermediate calculation, the 'weighted average redox standard' (SRO), to enable a value for mixtures of metabolites to be generated, the 'general mixture redox index' (GMOR). The indices were subjected to a proof-of-concept approach by making comparison to outcomes from multivariate analyses, i.e., PCA and HCA. Chemical analysis demonstrated that the essential oils were dominated by sesquiterpenes, constructed of 15 classes of compound (C-skeletons), and 4 C-skeletons were recognized in the monoterpene group, giving a total of 19. The variation of chemical profiles was distinct at different phenological stages, but stronger chemical variation was evident between day and night as compared to season. Furthermore, due to comprehensive sampling across different regions, nine chemotypes were recognized, including those previously reported. The SRO and GMRO indices demonstrate that phenological variation of chemistry is mainly an outcome of redox fluctuations in terpene biosynthesis, changing from day to night. These indices also corroborate that chemical diversity is increased with oxidative metabolism. Lastly, the current study demonstrates pronounced phenotypic plasticity in P. gaudichaudianum, which makes it a suitable candidate to help further our understanding of chemophenetics and chemical ecology.

Antileishmanial Activity of Flavones-Rich Fraction From Arrabidaea chica Verlot (Bignoniaceae).

Acknowledging the need of identifying new compounds for the treatment of leishmaniasis, this study aimed to evaluate, from in vitro trials, the activity of flavones from Arrabidaea chica against L. amazonensis. The chromatographic profiles of the hydroethanolic extract and a flavone-rich fraction (ACFF) from A. chica were determined by high-performance liquid chromatography coupled with a diode-array UV-Vis detector (HPLC-DAD-UV) and electrospray ionization mass spectrometry in tandem (LC-ESI-MS-MS). The flavones luteolin (1) and apigenin (2), isolated from chromatographic techniques and identified by Nuclear Magnetic Resonance of 1H and 13C, were also quantified in ACFF, showing 190.7 mg/g and apigenin 12.4 mg/g, respectively. The other flavones were identified by comparing their spectroscopic data with those of the literature. The in vitro activity was assayed against promastigotes and intramacrophagic amastigote forms of L. amazonensis. Cytotoxicity tests were performed with peritoneal macrophages of BALB/c mice. Nitrite quantification was performed with Griess reagent. Ultrastructural investigations were obtained by transmission electron microscopy. Anti-Leishmania assays indicated that the IC50 values for ACFF, apigenin, and luteolin were obtained at 40.42 ± 0.10 and 31.51 ± 1.13 µg/mL against promastigotes, respectively. ACFF and luteolin have concentration-dependent cytotoxicity. ACFF and luteolin also inhibited the intra-macrophagic parasite (IC50 3.575 ± 1.13 and 11.78 ± 1.24 µg/mL, respectively), with a selectivity index of 11.44 for ACFF. Promastigotes exposed to ACFF and luteolin exhibited ultrastructural changes, such as intense cytoplasm vacuolization and mitochondrial swelling. These findings data evidence the antileishmanial action of flavone-rich fractions of A. chica against L. amazonensis, encouraging further studies.

Essential Oils from Piper lhotzkyanum Kunth Leaves from Brazilian Atlantic Forest: Chemical Composition and Stability in Different Storage Conditions.

This work aimed to evaluate the impact of different storage conditions and light and temperature exposures on the visual aspect and chemical composition of the essential oil (EO) of Piper lhotzkyanum Kunth, obtained from leaves by hydrodistillation from a region of high altitude. For this purpose, aliquots of the EO were stored for up to 90 days (a) under a refrigerator condition of 5 ± 3°C, (b) under a long-term (LT) condition of 30 ± 2°C and 75 ± 5% relative humidity (RH) and an accelerated condition (AS) of 40 ± 2°C and 75 ± 5% RH, and (c) in a photostability test achieved in amber and colorless glass vials. The changes were monitored on days 0 (control), 60, and 90 for the refrigerator, LT, and AS conditions. All EO chemical analyses were assessed by GC-FID and GC-MS for quantification and identification, respectively. It is reported, for the first time, that the EO of P. lhotzkyanum is rich in the sesquiterpenes ß-elemene and α-zingiberene. No significant changes in the EO was observed, revealing a minimal impact of temperature on the sample at the different storage conditions. However, there was a change in the content of α-zingiberene to bicyclogermacrene after exposure to light. The visual appearance of the samples was altered for all test conditions except the refrigerator condition. These results can potentially contribute to the product development of a bioactive EO from leaves of P. lhotzkyanum, a sesquiterpene rich natural material.

The Influence of Anthocyanidin Profile on Antileishmanial Activity of Arrabidaea chica Morphotypes.

Arrabidaea chica Verlot (crajiru) is a plant used in folk medicine as an astringent, anti-inflammatory, wound healing and to treat fungal and viral diseases such as measles chickenpox and herpes. Arrabidaea chica has several morphotypes recognized but little is known about its chemical variability. In the present study the anthocyanidin profile of A. chica morphotypes collected in two seasons (summer and winter) have been examined and their activity against Leishmania infection compared. High-performance liquid chromatography coupled to a diode-array detector (HPLC-DAD-UV) and by tandem mass spectrometry with electrospray ionization (ESI-MS/MS) were used for anthocyanidin separation and identification. Antileishmanial activity was measured against promastigote forms of Leishmania amazonensis. Multivariate analysis, principal component analysis (PCA) and Pearson's correlation were performed to classify morphotypes accordingly to their anthocyanidin profile. The presence of 6,7,3',4'-tetrahydroxy-5-methoxyflavylium (3'-hydroxy-carajurone) (1), carajurone (2), 6,7,3'-trihydroxy-5,4'-dimethoxy-flavylium (3'-hydroxy-carajurin) (3) and carajurin (4), and three unidentified anthocyanidins were detected. Two different groups were recognized: group I containing 3'-hydroxy-carajurone; and group II with high content of carajurin. Among anthocyanidins identified in the extracts, only carajurin showed significant statistical correlation (p = 0.030) with activity against L. amazonensis. Carajurin could thus be considered as a pharmacological marker for the antileishmanial potential of the species.

Treatment of anal fistula with Baccharis dracunculifolia extract. Experimental study in rats. / Tratamento da fístula anal com extrato de Baccharis dracunculifolia. Estudo experimental em ratos.

OBJECTIVE: to evaluate the efficacy of Baccharis dracunculifolia extract in the treatment of anal fistulas in rats. METHODS: twenty male Wistar rats were submitted to anal fistula and, after 30 days, were divided into three groups: Control Group, with five animals; Carbopol Group, with five animals; and Baccharis dracunculifolia Group, with ten animals. In the Control Group, no treatment was performed. In the Carbopol Group, a daily infusion of Carbopol was performed for 30 days. In the Baccharis dracunculifolia Group, a daily infusion of Carbopol plus Baccharis dracunculifolia extract was performed for 30 days. Specimens were taken for histological analysis after euthanasia. RESULTS: there was no complete closure of the fistulous tract in any of the animals. The mean area of the remaining tract was of 847.2µm2, 565.6µm2 and 372.7µm2, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.001). The mean of the inflammatory process score was of 2.4, 2.4, and 2.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.285), while the mean values of vascular congestion were of 1.6, 1.4, and 1.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.031). CONCLUSION: Baccharis dracunculifolia extract was able to reduce the lumen of the fistulous tracts and the degree of vascular congestion, without, however, reducing the local inflammatory process or totally closing the fistulous tracts.

OBJETIVO: avaliar a eficácia do extrato de Baccharis dracunculifolia no tratamento de fístulas anais em ratos. MÉTODOS: vinte ratos Wistar machos foram submetidos à confecção de fístula anal e, após 30 dias, foram distribuídos em três grupos: Grupo Controle, com cinco animais; Grupo Carbopol, com cinco animais; e Grupo Baccharis dracunculifolia, com dez animais. No Grupo Controle, não se realizou nenhum tratamento. No Grupo Carbopol, realizou-se infusão diária de carbopol, e no Grupo Baccharis dracunculifolia, infusão de extrato de Baccharis dracunculifolia com carbopol, ambos por 30 dias. Foram retirados espécimes para análise histológica após a eutanásia. RESULTADOS: não houve fechamento completo do trajeto fistuloso em nenhum dos animais. A média da área do trajeto resultante foi de 847,2µm2, 565,6µm2 e 372,7µm2, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,001). A média do escore de processo inflamatório foi de 2,4, 2,4 e 2,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,285), enquanto a média de congestão vascular foi de 1,6, 1,4 e 1,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,031). CONCLUSÃO: o extrato de Baccharis dracunculifolia foi capaz de reduzir o lúmen dos trajetos fistulosos e a congestão vascular, sem reduzir, no entanto, o processo inflamatório local ou fechar totalmente os trajetos fistulosos.

Tratamento da fístula anal com extrato de Baccharis dracunculifolia. Estudo experimental em ratos / Treatment of anal fistula with Baccharis dracunculifolia extract. Experimental study in rats

RESUMO Objetivo: avaliar a eficácia do extrato de Baccharis dracunculifolia no tratamento de fístulas anais em ratos. Métodos: vinte ratos Wistar machos foram submetidos à confecção de fístula anal e, após 30 dias, foram distribuídos em três grupos: Grupo Controle, com cinco animais; Grupo Carbopol, com cinco animais; e Grupo Baccharis dracunculifolia, com dez animais. No Grupo Controle, não se realizou nenhum tratamento. No Grupo Carbopol, realizou-se infusão diária de carbopol, e no Grupo Baccharis dracunculifolia, infusão de extrato de Baccharis dracunculifolia com carbopol, ambos por 30 dias. Foram retirados espécimes para análise histológica após a eutanásia. Resultados: não houve fechamento completo do trajeto fistuloso em nenhum dos animais. A média da área do trajeto resultante foi de 847,2µm2, 565,6µm2 e 372,7µm2, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,001). A média do escore de processo inflamatório foi de 2,4, 2,4 e 2,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,285), enquanto a média de congestão vascular foi de 1,6, 1,4 e 1,1, respectivamente, nos Grupos Controle, Carbopol e Baccharis dracunculifolia (p=0,031). Conclusão: o extrato de Baccharis dracunculifolia foi capaz de reduzir o lúmen dos trajetos fistulosos e a congestão vascular, sem reduzir, no entanto, o processo inflamatório local ou fechar totalmente os trajetos fistulosos.

ABSTRACT Objective: to evaluate the efficacy of Baccharis dracunculifolia extract in the treatment of anal fistulas in rats. Methods: twenty male Wistar rats were submitted to anal fistula and, after 30 days, were divided into three groups: Control Group, with five animals; Carbopol Group, with five animals; and Baccharis dracunculifolia Group, with ten animals. In the Control Group, no treatment was performed. In the Carbopol Group, a daily infusion of Carbopol was performed for 30 days. In the Baccharis dracunculifolia Group, a daily infusion of Carbopol plus Baccharis dracunculifolia extract was performed for 30 days. Specimens were taken for histological analysis after euthanasia. Results: there was no complete closure of the fistulous tract in any of the animals. The mean area of the remaining tract was of 847.2µm2, 565.6µm2 and 372.7µm2, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.001). The mean of the inflammatory process score was of 2.4, 2.4, and 2.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.285), while the mean values of vascular congestion were of 1.6, 1.4, and 1.1, in the Control Group, Carbopol Group, and Baccharis dracunculifolia Group, respectively, (p=0.031). Conclusion: Baccharis dracunculifolia extract was able to reduce the lumen of the fistulous tracts and the degree of vascular congestion, without, however, reducing the local inflammatory process or totally closing the fistulous tracts.

The effect of Sebastiania hispida gel on wound model infected by methicillin resistant Staphylococcus aureus.

Medicinal plants are an alternative for the treatment of infected wounds. The objective of this study was to evaluate the efficacy of Sebastiania hispida in an animal model with a wound infected by Staphylococcus aureus. The crude ethanol extract (ExtEtOH) of S. hispida underwent phytochemical analysis, quantification of metabolites and antibacterial activity analysis performed using S. aureus. Wistar rats were used to test healing activity, and the groups evaluated comprised gels of ExtEtOH at the concentrations 0.2 and 2% compared with control groups. Animals were inoculated with the bacteria S. aureus resistant to methicillin. The treatment periods were of 3 and 21 days. Macroscopic and microscopic analysis were conducted and data were submitted to analysis of variance (p < 0.05). Phytochemical and quantification analysis indicated that phenolic compounds and flavonoids are the major constituents, followed by tyterpenes. ExtEtOH 0.2% was the most effective gel against the growth of strains of S. aureus. Histological wound and regression analysis showed that ExtEtOH gels (0.2% and 2%) were similar and effective in promoting wound healing. In the quantification of collagen fibers, the animals from all groups showed a high amount of thick collagen fibers. Thus, ExtEtOH gels based on the shoots of S. hispida can be used for the treatment of infected wounds as a complementary therapy for infected wound closure and further assays are required with other means. The healing effectiveness may be due to the high content of phenolics, flavonoids and triterpenes.

Identification of a Critical Window for Ganciclovir-Induced Disruption of Testicular Development in Rats.

Ganciclovir (GCV) has been implicated in the development of testicular alterations. Exposure on gestational day (GD) 10 in rats induced permanent effects, including focal reduction or absence of germ cells (Sertoli cell-only tubules). Because the timing of exposure can be critical for testicular effects, we exposed rat dams to 300 mg/kg GCV (3 100 mg/kg subcutaneous injections) on GD10, 14 and 19, when germ cells have high rates of migration, proliferation and are mitotically quiescent, respectively. Males exposed to GCV in utero on GD10 and 14 were evaluated for androgenization markers, serum and fecal androgens, and testicular histomorphometry at adulthood. Double-labeling immunofluorescence for DAZL and Ki67 were used to assess gonocytes number and the proliferative activity of germ and somatic cells in fetal testes on GD15 and 20, ie, 24 h after GCV exposure. Adult rats exposed on GD14 showed delayed puberty onset, despite normal androgen levels. Also, there was a 50% reduction in testicular weight and about 30% of seminiferous tubules lacking germ cells. Effects on GD10 animals were less pronounced. In the fetal testis, the number of gonocytes was reduced by 50% in rats exposed on GD14, but normal in GD19 fetuses. GCV also reduced Sertoli cell proliferation immunolabeling in GD19 fetuses and Sertoli cell number in adults. In conclusion, GCV toxicity on germ cells seems to be linked to their proliferation rate and GD14 is a critical window in rats, when GCV exposure causes an acute massive loss of germ cells that persists until adulthood.

Isolation of a larvicidal compound from Piper solmsianum C.DC. (Piperaceae).

The Aedes aegypti mosquito is one of the major vectors of arboviruses. These diseases have re-emerged and the insecticides used nowadays are toxic to mammals and environment and have only been effective in the short-term. In this context, natural products are an alternative. The genus Piper has many active compounds against arthropods, including neolignans. The present study evaluated the larvicidal potential of the n-hexanic extract of Piper solmsianum and eupomatenoid-6, identified by GC-MS and NMR techniques, from this extract against Ae. aegypti. The crude extract (100 µg/mL) killed 80% and 98.3% of larvae in the first and third day, respectively. Eupomatenoid-6 exhibited LD50 of 19.33 µM and LD90 of 28.68 µM and was then assayed in human fibroblast cells (MRC5), showing an IC50 of 39.30 µM with estimated LD50 of 42.26 mmol/kg. Our results indicate eupomatenoid-6 as a potent insecticide with relatively low toxicity for mammals.