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Abstract: The herbivory activity of the bordered patch larvae (Chlosyne lacinia, Lepidoptera) on leaves of a Brazilian population of Tithonia diversifolia and the antifeedant potential of its leaf rinse extract were investigated. The caterpillars fed only on the adaxial face, where the density of glandular trichomes is very low, and avoided the abaxial face, which contains high levels of trichomes. Deterrent activity against the larvae was observed in leaf discs treated with leaf rinse extract at concentrations of 1–5% of fresh leaf weight. High-performance liquid chromatography (HPLC) analysis indicated that sesquiterpene lactones are the main constituents of the glandular trichomes. Dichloromethane rinse extracts of the leaves and inflorescences were chemically investigated, and 16 compounds were isolated and identified: 14 sesquiterpene lactones, a flavonoid and a diterpenoid. In this study, five sesquiterpene lactones are described for the first time in the genus, including two lactones, one of which has an unusual seco-guaianolide skeleton. Our findings indicate that the caterpillars avoid the sesquiterpene-lactone-rich glandular trichomes, and provide evidence for the antifeedant activity of the dichloromethane leaf rinse extract. In addition, a study of the seasonal variation of the main constituents from the leaf surface throughout a year demonstrated that a very low qualitative but a very high quantitative variation occurs. The highest level of the main metabolite tagitinin C was observed between September and October and the lowest was from March to June, the later corresponding to the period of highest infestation by the larvae. [Copyright &y& Elsevier]

Background/Objectives: Brazilian red propolis has attracted attention for its pharmacological properties. However, signs of toxicity were recently observed in long-term studies using the hydroalcoholic extract of red propolis (RPHE), likely due to polyprenylated benzophenones. This study aimed to develop a benzophenone-free red propolis extract (BFRP) and validate an HPLC-PDA method to quantify its main constituents: isoliquiritigenin, vestitol, neovestitol, medicarpine, and 7-O-methylvestitol. Methods: BFRP's toxicity was assessed in zebrafish larvae through a vibrational startle response assay (VSRA) and morphological analysis. Genotoxicity was evaluated using the micronucleus test in rodents, and the extract's effects on chemically induced preneoplastic lesions in rat colon were studied. An HPLC-PDA method was used to quantify BFRP's main compounds. Results: BFRP primarily contained vestitol (128.24 ± 1.01 μg/mL) along with isoliquiritigenin, medicarpin, neovestitol, and 7-O-methylvestitol. Zebrafish larvae exposed to 40 µg/mL of BFRP exhibited toxicity, higher than the 10 µg/mL for RPHE, though no morphological differences were found. Fluorescent staining in the notochord, branchial arches, and mouth was observed in larvae treated with both BFRP and RPHE. No genotoxic or cytotoxic effects were observed up to 2000 mg/kg in rodents, with no impact on hepatotoxicity or nephrotoxicity markers. Chemoprevention studies showed a 41.6% reduction in preneoplastic lesions in rats treated with 6 mg/kg of BFRP. Conclusions: These findings indicate that BFRP is a safe, effective propolis-based extract with potential applications for human health, demonstrating reduced toxicity and chemopreventive properties. [ABSTRACT FROM AUTHOR]

Background/Objectives: Leishmaniasis, a neglected disease caused by Leishmania spp. including L. amazonensis, urgently requires new treatments. Polyalthic acid (PA), a natural diterpene from Copaifera spp., has previously demonstrated significant antiparasitic potential. This study evaluated the leishmanicidal effects of polyalthic acid (PA), alone and with amphotericin B (AmpB), on L. amazonensis promastigote and amastigote forms. Results: PA showed significant activity against promastigotes, with 50% effective concentration (EC50) values of 2.01 μM at 24 h and an EC50 of 3.22 μM against amastigotes after 48 h. The PA and AmpB combination exhibited a synergistic effect on both forms without inducing cytotoxicity or hemolysis. Morphological changes in promastigotes, including vacuole formation and cell rounding, were more pronounced with the combination. Conclusions: These findings suggest that PA and AmpB together could form a promising new treatment strategy against Leishmania infections, offering enhanced efficacy without added toxicity. [ABSTRACT FROM AUTHOR]

The technologies used to produce the different dosage forms of propolis can selectively affect the original propolis compounds and their biological activities. The most common type of propolis extract is hydroethanolic. However, there is considerable demand for ethanol-free propolis presentations, including stable powder forms. Three propolis extract formulations were developed and investigated for chemical composition and antioxidant and antimicrobial activity: polar propolis fraction (PPF), soluble propolis dry extract (PSDE), and microencapsulated propolis extract (MPE). The different technologies used to produce the extracts affected their physical appearance, chemical profile, and biological activity. PPF was found to contain mainly caffeic and p-Coumaric acid, while PSDE and MPE showed a chemical fingerprint closer to the original green propolis hydroalcoholic extract used. MPE, a fine powder (40% propolis in gum Arabic), was readily dispersible in water, and had less intense flavor, taste, and color than PSDE. PSDE, a fine powder (80% propolis) in maltodextrin as a carrier, was perfectly water-soluble and could be used in liquid formulations; it is transparent and has a strong bitter taste. PPF, a purified solid with large amounts of caffeic and p-Coumaric acids, had the highest antioxidant and antimicrobial activity, and therefore merits further study. PSDE and MPE had antioxidant and antimicrobial properties and could be used in products tailored to specific needs. [ABSTRACT FROM AUTHOR]

Rheumatoid arthritis is a systemic inflammatory autoimmune disease with prevalence estimated at 0.5% to 1% of the population. As one of the treatment routes of rheumatoid arthritis is based on the use of nonsteroidal anti-inflammatory drugs, the use of natural products with anti-inflammatory potential becomes relevant. Brown propolis has several biological properties, including immunomodulatory and anti-inflammatory effects. Thus, the present study evaluated the therapeutic efficacy of the crude extract of brown propolis from Araucaria sp. through experimental models of analgesia, anti-inflammatory activity, and rheumatoid arthritis. Hyperalgesia was evaluated by mechanical and thermal sensitivity. Anti-inflammatory activity was evaluated by plantar volume, cell migration and NF-kB expression in carrageenan-induced paw oedema. In collagen-induced rheumatoid arthritis, it was evaluated by mechanical and thermal nociception on the plantar surface, and mechanical nociception in the femorotibial and caudal joints, evaluation of plantar volume, radiography, weight gain and biochemical profile. The results demonstrated that the oral administration of brown propolis can modulate the course of rheumatoid arthritis, and it can inhibit pain through the modulation of mechanical sensitivity. The anti-arthritic effect of propolis may be due to its anti-inflammatory capacity, which includes inhibiting oedema formation, cell migration, and NF-kB expression, as well as preserving joint space and normalizing urea levels. This was an animal model study. Therefore, brown propolis should be evaluated in studies of human RA to determine efficacy. [ABSTRACT FROM AUTHOR]

Helicobacter pylori is a Gram-negative, microaerophilic, curved-rod, flagellated bacterium commonly found in the stomach mucosa and associated with different gastrointestinal diseases. With high levels of prevalence worldwide, it has developed resistance to the antibiotics used in its therapy. Brazilian red propolis has been studied due to its biological properties, and in the literature, it has shown promising antibacterial activities. The aim of this study was to evaluate anti-H. pylori from the crude hydroalcoholic extract of Brazilian red propolis (CHEBRP). For this, in vitro determination of the minimum inhibitory and bactericidal concentration (MIC/MBC) and synergistic activity and in vivo, microbiological, and histopathological analyses using Wistar rats were carried out using CHEBRP against H. pylori strains (ATCC 46523 and clinical isolate). CHEBRP presented MIC/MBC of 50 and 100 μg/mL against H. pylori strains (ATCC 43526 and clinical isolate, respectively) and tetracycline MIC/MBC of 0.74 µg/mL. The association of CHEBRP with tetracycline had an indifferent effect. In the stomach mucosa of rats, all treatments performed significantly decreased the number of H. pylori, and a concentration of 300 mg/kg was able to modulate the inflammatory response in the tissue. Therefore, CHEBRP showed promising anti-H. pylori in in vitro and in vivo assays. [ABSTRACT FROM AUTHOR]

As surgical site infection can prolong the patient's hospital stay, increase morbidity and mortality rates and medical and hospital expenses, justifying the search for new effective and low-cost antiseptic drugs is justifiable. Thus, the objective of the present study was to analyze, by direct clinical and microbiological examination, the topical antiseptic effect of a solution based on polyhexamethylene hydrochloride guanidine (PHMGH) at 0.5% on the intact skin surface of sheep and cats, aiming at its use in operative field (previous and definitive antisepsis), compared to 0.5% alcoholic chlorhexidine and association with this. The topical solutions did not cause any skin damage, regardless of species. The PHMGH demonstrated an antiseptic effect equivalent to chlorhexidine, however, the association between PHMGH and alcoholic chlorhexidine was more effective compared to isolated products. It is concluded that the topical products tested were harmless to the skin surface; PHMGH can be a preventive and less expensive option in skin antisepsis and that the association between PHMGH and alcoholic chlorhexidine was more effective in microbiological control when compared to the isolated products tested, however, more research will be essential to investigate the potentiation of these, as well as the performance of PHMGH in the presence of body fluids. [ABSTRACT FROM AUTHOR]

The genus Absidia is widely used in the biotransformation of different classes of natural products. This study evaluates the ability of the Absidia coerulea 3A9 marine derived strain isolated from the ascidian Distaplia stilyfera to perform biotransformations by conducting assays with (–)-cubebin, as substrate. The experiment was optimized using the experimental design proposed by Plackett–Burman for seven factors and eight experiments, to establish the biotransformation conditions that would allow maximum production of biotransformed dibenzylbutyrolactone (–)-hinokinin. An analytical method based on Reverse-Phase-High Performance Liquid Chromatography (RP-HPLC) was developed to quantify the fungal biotransformation product. The factor that influenced the (–)-hinokinin peak area the most positively was the percentage of seawater (%seawater) given that its %relative standard deviation (%RSD) showed a 32.92% deviation from the real value. [ABSTRACT FROM AUTHOR]

Propolis comprises a complex resinous product composed of plant's parts or exudates, pollen, bee wax, and enzymes. Brazilian brown propolis from Araucaria sp displays several biological activities. Considering the lack of validated analytical methods for its analysis, we are reporting the development of a validated high‐performance liquid chromatography with photodiode array detector method to analyze Araucaria brown propolis. The crude propolis were extracted and chromatographed, furnishing six main diterpenes. The isolated standards were used to draw the analytical curves, allowing the studies of selectivity, precision, accuracy, recovery, robustness, the determination of limits of detection and limits of quantification. The mobile phase consisted of 0.1% acetic acid in water and acetonitrile, using an octadecylsilane column, 1 mL/min flow rate and detection at 200 or 241 nm. Relative standard deviation values obtained for intra‐day and inter‐day precision were lower than 4% for all diterpenes. From the five parameters for robustness, wavelength detection and flow rate were the critical ones. Limits of detection and quantification ranged from 0.808 to 10.359 μg/mL and from 2.448 to 31.392 μg/mL, respectively. The recoveries were between 105.03 and 108.13%, with relative standard deviation values around 5.0%. The developed method is precise, sensitive, and reliable for analyzing Araucaria brown propolis. [ABSTRACT FROM AUTHOR]

Baccharin is one of the major compounds found in Brazilian green propolis and its botanical source, Baccharis dracunculifolia. Considering the biological effects of propolis and B. dracunculifolia, this study aims to evaluate the analgesic and anti-inflammatory potential of baccharin. The neurodepressor potential was performed by the open field test, analgesia by mechanical stimulation with Dynamic Plantar Aesthesiometer, and by thermal stimulation with Hargreaves apparatus. In addition, the anti-inflammatory potential was achieved by the paw edema assay, histopathological evaluation, and NF-kB expression. Doses of 2.5, 5, and 10 mg/kg of baccharin were evaluated. After euthanasia, plantar tissue was collected and prepared for histology. As a result, analgesic activity was observed at a dose of 10 mg/kg of baccharin in thermal stimulation under an inflammatory process and anti-inflammatory potential at a dose of 5 mg/kg of baccharin from the second hour in the paw edema test. A decrease in cellular infiltrate and down-modulation of NF-kB, besides the reduction of edema in the histopathology was observed. There was no evidence of kidney and liver toxicity and neurodepressive potential at the doses tested. Thus, baccharin has a promising anti-inflammatory effect possibly associated with antiedematogenic activity by inhibiting mediators such as prostaglandins, inhibiting the migration of polymorphonuclear cells, and modulating NF-kB expression. [Display omitted] [ABSTRACT FROM AUTHOR]

Helicobacter pylori is often related to severe clinical manifestations of infections and may increase an individual's chance of developing stomach cancer. We are reporting the in vitro anti-H. pylori effects of several Copaifera spp. derivatives such as oleoresins, hydroalcoholic crude extracts from the leaves, and isolated compounds, by determining their Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values. It was obtained promising MIC (MBC) values, ranging from 6.25 (12.5) to 100 (400) μg/mL, from 12.5 (25) to 100 (100) μg/mL, and from 3.12 (3.12) to 100 (100) μg/mL respectively, for Copaifera spp. oleoresins, leave crude hydroalcoholic extracts and isolated compounds. Therefore, all Copaifera spp. derivatives displayed promising antibacterial activities against H. pylori. [ABSTRACT FROM AUTHOR]

Invasion of periodontal tissues by Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans can be associated with aggressive forms of periodontitis. Oleoresins from different copaifera species and their compounds display various pharmacological properties. The present study evaluates the antibacterial and antivirulence activity of oleoresins obtained from different copaifera species and of ten isolated compounds against two causative agents of periodontitis. The following assays were performed: determination of the minimum inhibitory concentration (MIC), determination of the minimum bactericidal concentration (MBC), and determination of the antibiofilm activity by inhibition of biofilm formation and biofilm eradication tests. The antivirulence activity was assessed by hemagglutination, P. gingivalis Arg-X and Lis-X cysteine protease inhibition assay, and A. actinomycetemcomitans leukotoxin inhibition assay. The MIC and MBC of the oleoresins and isolated compounds 1, 2, and 3 ranged from 1.59 to 50 μg/mL against P. gingivalis (ATCC 33277) and clinical isolates and from 6.25 to 400 μg/mL against A. actinomycetemcomitans (ATCC 43717) and clinical isolates. About the antibiofilm activity, the oleoresins and isolated compounds 1, 2, and 3 inhibited biofilm formation by at least 50% and eradicated pre-formed P. gingivalis and A. actinomycetemcomitans biofilms in the monospecies and multispecies modes. A promising activity concerning cysteine protease and leucotoxin inhibition was also evident. In addition, molecular docking analysis was performed. The investigated oleoresins and their compounds may play an important role in the search for novel sources of agents that can act against periodontal pathogens. [ABSTRACT FROM AUTHOR]

Objective: Evaluation of the healing and toxicological effects of Copaifera duckei Dwyer oleoresin (CDO). Materials and Methods: Rodents with skin lesions were divided into nine groups, including daily treatments with 1, 3 and 10% CDO, collagenase, antibiotic ointment and control groups, for 14 days. Results: Treatment with 10% CDO reduced skin edema and hyperplasia, demonstrating anti-inflammatory effect of the oil. Reduction in the wound area was observed, indicating the healing effect of CDO. Histopathological analysis showed increases in angiogenesis and re-epithelialization in animals treated with the highest concentration. On the other hand, no alterations in ulcerations, inflammatory infiltrate, hemorrhage, congestion, degeneration, percentage of collagen fibers, number of cells stained with antimacrophage migration inhibitory factor, or density of area stained with anti-collagen I and III were found. Toxicogenetic analysis revealed no differences in micronucleus frequencies or in the ratio of polychromatic erythrocytes to total erythrocytes between treated and negative control, demonstrating the absence of genotoxicity and cytotoxicity, respectively. There was no difference in levels of liver enzymes among groups, indicating the absence of hepatotoxicity. Conclusion: Formulations of CDO exerted beneficial effects on the stages of cutaneous wound healing and are promising options for the treatment of wounds. [ABSTRACT FROM AUTHOR]

• Diterpenes are a source of potential leading compounds to treat schistosomiasis. • Derivatives were obtained by fungal transformation with Aspergillus and Cunninghamella. • Three precursors and ten diterpene derivatives were evaluated against Schistosoma mansoni. • 7α-acetoxy- ent -kaur-16-en-19-oic acid (KA5) and ent -hardwickiic acid (HA) were the most active ones. • KA5 and HA are potential candidates for schistosomicidal drug discovery. Schistosomiasis continues to be a huge challenge for researchers, pharmaceutical companies, and governments in developing countries. Diterpene compounds are good source for the development of novel potential leading compounds to treat schistosomiasis. We are reporting herein the schistosomicidal activity of ent -kaurenoic acid, ent -copalic acid, ent -hardwickiic acid, isolated from oleoresins of Copaifera spp, and of their derivatives obtained by fungal transformation with strains of Aspergillus fumigatus , A. terreus , A. phoenicis and A. ochraceus , and of Cunninghamella echinulata e C. elegans. The in vitro antiparasitical assays were performed using adult worm pairs of Schistosoma mansoni for the evaluation of the worm pairing, egg production, and eggs development. Ten kaurane, labdane and clerodane-type diterpenes were obtained by fungal transformation and 7α-acetoxy- ent -kaur-16-en-19-oic acid and ent -hardwickiic acid were the most active ones by causing mortality of 100 % of the parasites within 24 h (concentrations of 100.0 and 200.0 μM) and displaying respective IC 50 values for 24, 48 and 72 h of 56.7, 37.6 and 29.2 μM and 29.6, 30.8 and 25.7 μM. Additionally, 7α-acetoxy- ent -kaur-16-en-19-oic acid and ent -hardwickiic acid highly reduced the number of laid eggs at 6.25 and 12.5 μM, respectively. These diterpenes should be further investigated as potential candidates for antiparasitic drug discovery. [ABSTRACT FROM AUTHOR]

The rise in antifungal resistance and side effects of conventional treatments drive the search for innovative therapies like Photodynamic Therapy (PDT). This study explored the efficacy of PDT mediated by gutiferone, an isolated compound from red propolis, for candidiasis treatment. Multiple evaluation methods were employed, including determining the minimum inhibitory concentration (MIC) via broth microdilution, quantifying biomass using crystal violet detachment, and cell counting through total plate count. PDT mediated by gutiferone was also assessed in five groups of mice, followed by histopathological examination and agar plating of lingual tissue samples. Among the seven Candida species tested, gutiferone displayed efficacy against C. albicans, C. glabrata , and C. tropicalis , with MIC values of 1000 μg/mL. In C. tropicalis biofilms, exposure to gutiferone led to a reduction of 1.61 Log10 CFU/mL. PDT mediated by gutiferone achieved an average reduction of 3.68 Log10 CFU/mL in C. tropicalis biofilm cells, underscoring its potent fungicidal activity. Histopathological analysis revealed fungal structures, such as pseudohyphae and hyphae, in infected groups (G2) and irradiated mice. In contrast, groups treated with gutiferone or subjected to gutiferone-assisted PDT (G5) exhibited only few blastoconidia. Furthermore, CFU/mL assessments in lingual tissue post-treatment demonstrated a significantly lower count (0.30 Log10 CFU/mL) in the G5 group compared to G2 (2.43 Log10 CFU/mL). These findings highlight the potential of PDT mediated by gutiferone as a promising alternative for managing denture stomatitis. Future research and clinical investigations offer the promise of validating its clinical applicability and improving outcomes in the treatment of oral candidiasis. • Antifungal resistance drives Photodynamic Therapy exploration. • Gutiferone's efficacy against Candida species. • Potent fungicidal activity in Candida tropicalis. • Histopathological improvement in group treated simultaneously with photodynamic therapy and gutiferone. • Gutiferone-assisted Photodynamic Therapy for oral candidiasis. [ABSTRACT FROM AUTHOR]

The search for new antibacterial agents and a better comprehension of substances with antimicrobial behavior is mandatory nowadays due to the serious public health problem of infection diseases. In the present work, 30 diterpenes were studied, with 2 natural derivatives, named ent-16-kauren-19-oic acid and ent-pimara-8(14),15-dien-19-oic acid, and 28 semi-synthetic derivatives. The natural diterpenes were isolated from Mikania glomerata and Viguiera arenaria, respectively. All diterpenes were submitted to antimicrobial assays against six different Gram-positive microorganisms to better understand the structure–activity relationship of antimicrobial diterpenes. The semi-synthetic derivatives were all obtained from the two natural derivatives by structural modifications, mainly esterification reactions. Both natural derivatives, together with the derivative ent-8(14)-pimaren-19-oic acid, displayed the most relevant antibacterial activities, with minimal inhibitory concentration (MIC) values that were less than 10 μg mL–1 for most pathogens; thus, they were considered promising antimicrobial agents. Moreover, in light of the hypothesis of Urzúa and colleagues, several considerations about the structure–activity relationship of antimicrobial diterpenes could be stated. [ABSTRACT FROM AUTHOR]

Abstract Copaifera is a genus of large trees found in Brazil, mainly in Amazon forest, but also in Atlantic forest and cerrado biomes. It has also been found in other countries in South America. In Africa, it is found mainly in Congo, Cameroon, Guinea and Angola. Its oleoresin has been used in folk medicine in the treatment of numerous healthy disorders, such as urinary, respiratory, skin and inflammatory diseases, for which there are several studies corroborating its ethnopharmacological uses. It is also extensively employed in the pharmaceutical and cosmetic industries in the development of ointments, pills, soaps, perfumes, among others. Copaifera oleoresin contains mainly diterpenes, such as: kaurenoic acid, kaurenol, copalic acid, agathic acid, hardwiickic acid, polyalthic acid, and sesquiterpenes, comprising β -caryophyllene, caryophyllene oxide, α-copaene, α–humulene, γ-muurolene and β -bisabolol, among other compounds. On the other hand, Copaifera leaves contain mainly phenolic compounds, such as flavonoids and methylated galloylquinic acid derivatives. Therefore, considering the economic importance of Copaifera oleoresin, its ethnopharmacological uses, the need to develop new pharmaceuticals for the treatment of many diseases, as well as the pharmacological potential of the compounds found in Copaifera spp., it was undertaken a review covering mostly the last two decades on the distribution, chemistry, pharmacology, quality control and safety of Copaifera species. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Copaifera species are used in folk medicine for a wide variety of pharmacological properties. This paper reports the cytotoxic and genotoxic analyses of oleoresins and leaves extracts of Copaifera species: C. duckei , C. multijuga , C. paupera , C. pubiflora, C. reticulata and C. trapezifolia . Materials and methods In vitro assays were performed using Chinese hamster lung fibroblasts (V79 cells). The clonogenic efficiency and cytokinesis-block micronucleus assays were employed for the cytotoxicity and genotoxicity assessment, respectively. The mouse bone marrow micronucleus test was used for in vivo studies. Results The cytotoxicity results using the clonogenic efficiency assay showed IC 50 values ranging from 9.8 to 99.2 µg/mL for oleoresins and 66.4–721.5 for leaves extracts. However, no cytotoxic effect was observed in the in vivo studies. Additionally, the treatments with oleoresins and leaves extracts did not significantly increase the frequency of micronuclei in both in vitro and in vivo mammalian cells. The UPLC-MS/MS and CG/MS analyses of Copaifera oleoresins allowed the identification of 10 acid diterpenes and 11 major volatile sesquiterpenes. Leaves are rich in phenolic compounds including two flavonoid heterosides and 16 galloylquinic acid derivatives. Conclusions The oleoresins and leaves extracts of studied Copaifera species were not cytotoxic in vivo, as well as not genotoxic in both in vitro and vivo assays, under the experimental conditions used. Therefore, the obtained results should be sufficient to demonstrate the absence of significant genotoxic risk of these Copaifera products for human use in the evaluated concentrations range. [ABSTRACT FROM AUTHOR]

Dehydroabietic acid (DHA) is one of the main constituents of the resin that have antiprotozoal activity against Leishmania spp., but the leishmanicidal mechanism is unknown. The objective of the study was to investigate in vitro the leishmanicidal activity of the natural compound DHA against intracellular and extracellular forms of L. amazonensis and the mechanism of action involved. The antileishmanial activity of DHA was evaluated in vitro against promastigote forms of L. amazonensis by counting in Neubauer chamber. The morphological changes were observed by scanning electron microscopy and cell death mechanism by fluorescence assay using 2′,7′-dichlorofluorescein diacetate probe (H 2 DCFDA), tetramethylrhodamine ethyl ester (TMRE), annexin-V and propidium iodide (PI). The antiamastigote effect was observed by counting the number of amastigotes per macrophage and percentage of infected cells. In addition, reactive oxygen species (ROS) production, nitric oxide (NO), cytokines, free iron and total iron-binding capacity (TIBC), expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and ferritin were evaluated. DHA inhibited the proliferation of promastigotes at all times tested. The compound (IC50, 40 ± 0.1458 μg/mL) altered the morphology of the promastigote forms, caused mitochondrial depolarization, induced ROS production, increased phosphatidylserine exposure and caused loss of plasma membrane integrity. DHA also reduced the number of amastigotes and the percentage of infected macrophages by increasing ROS production, free iron and TIBC, and also caused downregulation of Nrf2 and ferritin expression. DHA was effective in the elimination of L. amazonensis both in its promastigote forms by apoptosis-like mechanisms and intracellular form by ROS production. [ABSTRACT FROM AUTHOR]

In view of the biological activities and growing therapeutic interest in oleoresin obtained from Copaifera multijuga, this study aimed to determine the genotoxic and antigenotoxic potential of this oleoresin (CMO) and its chemical marker, diterpene (-)-copalic acid (CA). The micronucleus (MN) assay in V79 cell cultures and the Ames test were used for in vitro analyses, as well as MN and comet assays in Swiss mice for in vivo analyses. The in vitro genotoxicity/mutagenicity results showed that either CMO (30,60, or 120 µg/ml-MN assay; 0.39-3.12 mg/plate-Ames test) or CA (2.42; 4.84, or 9.7 µg/ml-MN assay; 0.39-3.12 mg/plate-Ames test) did not induce a significant effect on the frequency of MN and number of revertants, demonstrating an absence of genotoxic and mutagenic activities, respectively, in vitro. In contrast, these natural products significantly reduced the frequency of MN induced by methyl metha- nesulfonate (MMS), and exerted a marked inhibitory effect against indirect-acting mutagens in the Ames test. In the in vivo test system, animals treated with CMO (6.25 mg/kg b.w.) exhibited a significant decrease in rate of MN occurrence compared to those treated only with MMS. An antigenotoxic effect of CA was noted in the MN test (1 and 2 mg/kg b.w.) and the comet assay (0.5 mg/kg b.w.). Data suggest that the chemical marker of the genus Copaifera, CA, may partially be responsible for the observed chemopreventive effect attributed to CMO exposure. [ABSTRACT FROM AUTHOR]

Background: Patients submitted to hemodialysis therapy are more susceptible to infection, especially to infection by Gram-positive bacteria. Various research works have attempted to discover new antimicrobial agents from plant extracts and other natural products. Methods: The present study aimed to assess the antibacterial activities of Copaifera duckei, C. reticulata, and C. oblongifolia oleoresins; sodium hypochlorite; and peracetic acid against clinical and environmental isolates recovered from a Hemodialysis Unit. The Minimum Inhibitory Concentration and the Fractionated Inhibitory Concentration Index were determined; the ability of the tested compounds/extracts to inhibit biofilm formation was evaluated by calculating the MICB50 and IC50. Results: C. duckei was the most efficient among the assayed Copaifera species, and its oleoresin was more effective than peracetic acid and sodium hypochlorite. Copaifera oleoresins and disinfectants did not act synergistically at any of the tested combinations. Certain of C. duckei oleoresin, peracetic acid, and sodium hypochlorite concentrations inhibited biofilm formation and eradicated 50% of the biofilm population. Conclusion: C. duckei oleoresin is a potential candidate for disinfectant formulations. Based on these results and given the high incidence of multi-resistant bacteria in hemodialysis patients, it is imperative that new potential antibacterial agents like C. duckei oleoresin, which is active against Staphylococcus, be included in disinfectant formulations. [ABSTRACT FROM AUTHOR]

Copaiba oil is widely used in medicine, but there are no reports regarding its application in ophthalmology. Therefore, the objective of this study was to evaluate the clinical, histopathological and toxicogenetic effects of eye drops containing 0.1 and 0.5% of Copaifera multijuga Hayne oil on superficial corneal ulcers induced with alkali in the left eye of rats. For histological analysis, the percent reduction in ulcers and thickness of the corneal epithelium and stroma were evaluated 48 and 72 h after ulcer induction. Additionally, neovascularization and polymorphonuclear infiltration were classified in the stroma. The bone marrow micronucleus test was used for toxicogenetic assessment. None of the animals exhibited clinical signs of immediate ocular discomfort after instillation and the eye drops were harmless to the ocular surface. There was a significant difference in percent ulcer reduction and corneal stroma thickness between animals treated with the C. multijuga eye drops and untreated animals with corneal injury and the negative control, respectively, suggesting a healing effect of the oleoresin. Analysis of the thickness of the corneal epithelium at the two time points showed that the eye drops formulated did not significantly reduce the damage caused by alkali. The same was observed for the treatments with the reference drugs. No difference in stromal neovascularization or inflammatory infiltration was observed between the treated groups. The toxicogenetic results revealed the absence of cytotoxicity and genotoxicity of the treatments. In conclusion, the C. multijuga eye drops did not cause damage to the ocular surface under the present experimental conditions and corneal epithelization was similar to the conventional treatments. These results indicate that eye drops containing C. multijuga oleoresin are a promising option for the treatment of superficial keratitis. [ABSTRACT FROM AUTHOR]

This paper is the first report on the in vitro effects of licochalcone A, a chalcone isolated from Glycyrrhiza inflate Batalin (Leguminosae), on Schistosoma mansoni adult worms. In vitro , licochalcone A afforded lethal concentrations for 50% of parasites (LC 50 ) of 9.12 ± 1.1 and 9.52 ± 0.9 μM against female and male adult worms, respectively, at 24 h. Additionally, the compound reduced the total number of S. mansoni eggs and affected the development of eggs produced by S. mansoni adult worms. Together, the results achieved after 24 h showed that licochalcone A was 55.7- and 53.3-fold more toxic to S. mansoni female and male adult worms than to Chinese hamster ovary fibroblasts cells, respectively. Treatment with licochalcone A elicited drastic changes in the tegument of S. mansoni adult worms, as well as mitochondrial alteration and chromatin condensation. Licochalcone A also increased the superoxide anion level and decreased the superoxide dismutase activity in S. mansoni adult worms. Overall, our results indicated that licochalcone A displays in vitro schistosomicidal activity. This effect may result from increased production of reactive oxygen species (ROS) induced by the action of licochalcone A. The resulting ROS could act on the S. mansoni tegument and membranes and help induce the death of S. mansoni adult worms. [ABSTRACT FROM AUTHOR]

Species of Copaifera genus (Fabaceae – Caesalpinoiodidaeae) produces an important commercial oleoresin that displays many medicinal properties. Copaifera oleoresins (COR) are composed mainly of a mixture of diterpenes and sequiterpenes, and the main reported acid diterpenes for this genus are kaurenoic, copalic, hardwickiic and polyaltic acids. An ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method was developed and validated for identification and quantification of nine acid diterpenes. The developed method was applied in the analyses of 10 authentic COR samples collected in the North and Southeast of Brazil and six commercial COR samples. Samples preparation consisted of simple dilution of oleoresins in methanol followed by filtration. Validation parameters of the method for nine acid diterpenes were satisfactory: selectivity/specificity was defined by retention time and MS/MS analyses for each analyte; generally all analytical curves presented r 2 > 0.99, Lack-of-fit test not significant and RSD < 20% for all concentration levels; limit of detection and limit of quantification were on the scale of nanogram per milliliter; inter- and intra-day precision and accuracy were adequate. Regarding the robustness, the method was sensible to small deliberate variations of temperature and additives to the mobile phase, such as formic acid and ammonium hydroxide. Results of 16 analyzed samples of COR showed qualitative and quantitative differences of acid diterpenes among all samples. The diterpenes ent -kaurenoic acid 1 , ent -polyalthic acid 3 , ent -copalic acid 5 and, ent -3- β -acetoxy copalic acid 9 were found with more frequency in COR analyzed samples. Additionally, the content of the acid diterpenes found in 16 Copaifera oleoresin samples was analyzed by Principal Component Analysis (PCA), suggesting a botanical origin for the commercial samples. The developed UPLC method was shown to be reliable for the analysis of acid diterpenes in commercial Copaifera oleoresins. [ABSTRACT FROM AUTHOR]

The goal of our study was to evaluate the effect of kaurenoic acid, obtained from copaiba oil resin, in gastric cancer (GC) and a normal mucosa of stomach (MNP01) cell lines. The compound was tested at concentrations of 2.5, 5, 10, 30 and 60 μg/mL. Comet and micronucleus assays were used to access its potential genotoxicity in vitro . Moreover, we evaluated the effect of kaurenoic acid in cell cycle progression and in the transcription of genes involved in the control of the cell cycle: MYC , CCND1 , BCL2 , CASP3 , ATM , CHK2 and TP53 . Kaurenoic acid induced an increase on cell DNA damage or micronucleus frequencies on GC cell lines in a dose-dependent manner. The GC and MNP01 cell lines entering DNA synthesis and mitosis decreased significantly with kaurenoic acid treatment, and had an increased growth phase compared with non-treated cells. The treatment induced apoptosis (or necrosis) even at a concentration of 2.5 μg/mL in relation to non-treated cells. GC cell lines presented reduced MYC , CCND1 , BCL2 and CASP3 transcription while ATM , CHK2 and TP53 increased in transcription in relation to non-treated cells, especially at a concentration above 10 μg/mL. The gene transcription in the MNP01 (non-treated non-cancer cell line) was designated as a calibrator for all the GC cell lines. In conclusion, our results showed that kaurenoic acid obtained from Copaifera induces DNA damage and increases the micronuclei frequency in a dose-dependent manner in GC cells, with a significant genotoxicity observed above the concentration of 5 μg/mL. Moreover, this compound seems to be able to induce cell cycle arrest and apoptosis in GC cells. [ABSTRACT FROM AUTHOR]

The diterpene kaurenoic acid (KA) has vasorelaxant, antimicrobial, anti-tumoural and anti-leishmanial effects. Semi-synthetic derivatives were obtained to achieve more satisfactory responses. The assessment of genotoxicity is part of the toxicological evaluation of therapeutic compound candidates. The present study investigated the cytotoxicity and genotoxicity of KA and its semi-synthetic derivatives methoxy kaurenoic acid (MKA) and kaurenol (KRN) using the CHO-K1 cell line. The cytotoxicity evaluation demonstrated that treatments with 200 and 400 μM KA reduced cellular proliferation to 36.5 and 4.43%, respectively, and that 100 and 200 μM KA reduced the survival fraction (SF) to 48.1 and 5.5%, respectively. MKA and KRN at concentrations of 400 μM reduced proliferation to 81 and 86.8%, respectively, while 100 and 200 μM KRN reduced the SF to 50%, and 200 μM MKA reduced the SF to 74%. No genotoxicity was observed for KA or MKA. However, 100 μM KRN increased the DNA damage index, as detected by comet assay, although a micronucleus assay did not confirm these data. The results demonstrated that KA and its semi-synthetic derivative MKA were not genotoxic when tested at noncytotoxic concentrations, but KRN was genotoxic at the highest concentration that was tested, as demonstrated by the comet assay. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the antibacterial activity of crude Brazilian red propolis (BRP) extract against anaerobic bacteria involved in primary endodontic infection. Additionally, we evaluate the cell viability and free radical production of human dental pulp fibroblasts (HDPF) in direct contact with mineral trioxide aggregate (MTA) and BRP. The Minimum Inhibitory Concentration, Minimum Bactericidal Concentration (MIC, MBC) and Minimum Inhibitory Concentration of Biofilm (MICB 50) of BRP against anaerobic endodontic pathogens were determined. HDPF were exposed to BRP10 (10 μg/mL), BRP50 (50 μg/mL), MTA extract (1:1, 1:2, 1:4 e 1:8), dimethyl sulfoxide 0.5% (DMSO), and cell culture medium (DMEM). The groups were tested for cell viability (MTT assay), and free radical production (reactive oxygen species - ROS, DCFH-DA probe and nitric oxide – NO, Griess reagent). The one-way ANOVA and Tukey's tests were employed at a significance level of 5%. MIC/MBC values of BRP performed antibacterial activity for Parvimonas micra (6.25/6.25 µg/mL), Fusobacterium nucleatum (25/25 µg/mL), Prevotella melaninogenica (50/100 µg/mL), Prevotella nigrescens (50/100 µg/mL), Prevotella intermedia (50/100 µg/mL), and Porphyromonas gingivalis (50/200 µg/mL). The MICB 50 values ranged from 1.56 to 50 μg/mL. BRP and MTA stimulated cell viability, emphasizing BRP10 (p = 0.007). Furthermore, it was observed that MTA 1:1, MTA 1:2, and BRP50 slightly increased ROS (p < 0.001) and NO production (p = 0.008, p = 0.007, and p < 0.001 respectively) compared to DMEM group. BRP exhibits good antibacterial activity against endodontic pathogens, and both BRP and MTA promote the viability of HDPF without increasing NO and ROS production. • The cytotoxicity findings demonstrated that BRP and MTA stimulated cell viability in all tested concentrations. • HDPF production of NO and ROS was maintained at basal levels in direct contact with MTA and BRP. • Higher cell viability values were found for BRP10, as well as it did not increase free radicals production. • BRP is a strong candidate for future studies aiming the design of new antimicrobial formulations in VPTs. [ABSTRACT FROM AUTHOR]

Leishmania amazonensis (L. amazonensis) infection can cause severe local and diffuse injuries in humans, a condition clinically known as American cutaneous leishmaniasis (ACL). Currently, the therapeutic approach for ACL is based on Glucantime, which shows high toxicity and poor effectiveness. Therefore, ACL remains a neglected disease with limited options for treatment. Herein, the in vitro antiprotozoal effect and mechanisms of the diterpene kaurenoic acid [ent-kaur-16-en-19-oic acid] (KA) against L. amazonensis were investigated. KA exhibited a direct antileishmanial effect on L. amazonensis promastigotes. Importantly, KA also reduced the intracellular number of amastigote forms and percentage of infected peritoneal macrophages of BALB/c mice. Mechanistically, KA treatment reestablished the production of nitric oxide (NO) in a constitutive NO synthase- (cNOS-) dependent manner, subverting the NO-depleting escape mechanism of L. amazonensis. Furthermore, KA induced increased production of IL-1β and expression of the inflammasome-activating component NLRP12. These findings demonstrate the leishmanicidal capability of KA against L. amazonensis in macrophage culture by triggering a NLRP12/IL-1β/cNOS/NO mechanism. [ABSTRACT FROM AUTHOR]

Copaifera spp oleoresins have been used in folk medicine for centuries; nevertheless, its immunomodulatory action has not been investigated. Thus, the goal of this study was to characterize different oleoresins and to verify their action on human monocytes regarding pro- and anti-inflammatory cytokine production (TNF-α and IL-10, respectively). The chemical composition of Brazilian Copaifera reticulata , Copaifera duckey and Copaifera multijuga oleoresins was analyzed by HPLC-MS. Cell viability was assessed by MTT method after incubation of cells with Copaifera spp. Noncytotoxic concentrations of oleoresins were incubated with human monocytes from healthy donors, and cytokine production was determined by ELISA. HPLC-MS analysis for terpenes allowed the identification of six diterpene acids and one sesquiterpene acid. Oleoresins exerted no cytotoxic effects on human monocytes. All oleoresins had a similar profile: LPS-induced TNF-α production was maintained by oleoresins, while a significant inhibitory action on IL-10 production was seen. Copaifera oleoresins seemed to exert an activator profile on human monocytes without affecting cell viability. Such effect may be due to the presence of either diterpene or sesquiterpene acids; however, further studies are necessary to determine the involvement of such compounds in Copaifera immunomodulatory effects. [ABSTRACT FROM AUTHOR]

Abstract: The pimarane-type diterpene, pimaradienoic acid (PA), is known for its diverse biological properties such as antimicrobial, anti-inflammatory and trypanocidal. A preliminary study was undertaken to investigate in vitro the free radical-scavenging potential of PA. In addition, the genotoxic potential of PA and its ability to modulate genotoxicity induced by doxorubicin (DXR) and methyl methanesulfonate (MMS) were studied in Chinese hamster lung fibroblasts (V79 cells) and in male Swiss mice using the comet and micronucleus assays. The DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay showed that PA exerted no antioxidant activity when compared to quercetin. The colony-forming assay using V79 cells showed that PA was cytotoxic at concentrations >5.0μg/mL. Therefore, concentrations of 0.625, 1.25, 2.5, and 5.0μg/mL were used for evaluation of the genotoxic and antigenotoxic potential of PA in V79 cells. For genotoxic and antigenotoxic assessment in Swiss mice, three PA doses were tested (20, 40, and 80mg/kg body weight) based on the solubility limit of the diterpene in dimethylsulfoxide and water. The in vitro results demonstrated that PA induced DNA damage at concentrations of 2.5 and 5.0μg/mL in the comet assay. However, no genotoxic effect was observed in the micronucleus test using V79 cells. In the in vivo evaluation of genotoxicity, a significant increase in the frequency of DNA damage was observed in hepatocytes of animals treated with the highest PA dose (80mg/kg) when compared to the control group, but this difference was not seen in the micronucleus test. Furthermore, PA significantly reduced the frequency of DXR- and MMS-induced micronuclei and extent of DNA damage in in vitro and in vivo test systems. [Copyright &y& Elsevier]

Copaifera species folkloric names are "copaíbas, copaibeiras, copaívas or oil stick", which are widely used in Brazilian folk medicine. Among all ethnopharmacological applications described for Copaifera spp oleoresins, their anti-inflammatory effect stands out. However, the knowledge of anti-inflammatory and antinociceptive properties of Copaifera pubiflora Benth is scarce. To investigate the cytotoxic, anti-inflammatory, and antinociceptive activities of C. pubiflora oleoresin (CPO), and its major compound ent -hardwickiic acid (HA). The phosphatase assay was used to evaluate the cytotoxicity of CPO and HA in three different cell lines. CPO and HA doses of 1, 3, and 10 mg/kg were employed in the biological assays. The assessment of motor activity was performed using open-field and rotarod tests. Anti-inflammatory activity of CPO and HA was assessed through luciferase assay, measurement of INF- γ , IL-1 β , IL-6, IL-10, and TNF- α in a multi-spot system with the immortalized cell line THP-1, zymosan-induced arthritis, and carrageenan-induced paw edema. Acetic acid-induced abdominal writhing and formalin tests were undertaken to evaluate the antinociceptive potential of CPO and HA. In addition, the evaluation using carrageenan was performed to investigate the effect of CPO in pain intensity to a mechanical stimulus (mechanical hyperalgesia), using the von Frey filaments. A tail-flick test was used to evaluate possible central CPO and HA actions. In the cytotoxicity evaluation, CPO and HA were not cytotoxic to the cell lines tested. CPO and HA (10 mg/kg) did not affect animals' locomotor capacity in both open-field and rotarod tests. In the luciferase assay, CPO and HA significantly reduced luciferase activity (p < 0.05). This reduction indicates a decrease in NF- κ B activity. HA and CPO decreased INF- γ , IL-1 β , IL-6, IL-10, and TNF- α at 24 and 72 h in the multi-spot system. In zymosan-induced arthritis, CPO and HA decreased the number of neutrophils in the joint of arthritic mice and the number of total leukocytes (p < 0.05). In experimental arthritis HA significantly decreased joint swelling (p < 0.05). CPO and HA also increased the mechanical threshold during experimental arthritis. HA and CPO significantly inhibited the carrageenan-induced paw edema, being the doses of 10 mg/kg the most effective, registering maximum inhibitions of 58 ± 8% and 76 ± 6% respectively, p < 0.05. CPO and HA reduced the nociceptive behavior in both phases of formalin at all tested doses. The highest doses tested displayed inhibitions of 87 ± 1% and 72 ± 4%, respectively, p < 0.001, in the first phase, and 87 ± 1% and 81 ± 2%, respectively, p < 0.001, in the second phase. Oral treatment of CPO and HA (1, 3, 10 mg/kg) significantly reduced the nociceptive response in acetic acid-induced abdominal writhings, and the 10 mg/kg dose was the most effective with maximum inhibitions of 86 ± 2% and 82 ± 1%, respectively, p < 0.001. Both HA and CPO significantly decreased the intensity of mechanical inflammatory hyper-nociception on carrageenan-induced hyperalgesia at all tested doses, and 10 mg/kg was the most effective dose with maximum inhibitions of 73 ± 5% and 74 ± 7%, respectively, p < 0.05.CPO increased the tail-flick latencies in mice, and concomitant administration of naloxone partially reduced its effect. CPO and HA may inhibit the production of inflammatory cytokines by suppressing the NF- κ B signaling pathway, resulting in anti-inflammatory and antinociceptive activities. Image 1 [ABSTRACT FROM AUTHOR]

• A RP-HPLC analytical method is reliable to analyze chemical markers of Brazilian red propolis. • Vestitol, medicarpin, and neovestitol are major compounds on Brazilian red propolis samples. • Rainy season increase phenolic compounds concentration on Brazilian red propolis. • Brazilian red propolis chemical markers are present D. ecastaphyllum stems and resin. Propolis is a natural product produced from the interaction between bees and plants. Brazilian red propolis results from Apis mellifera collection of resins from two plant species, being Dalbergia ecastaphyllum (L.) Taub, Fabaceae, the primary botanical source, containing isoflavonoids and other characteristic phenolic compounds. Several biological activities of Brazilian red propolis and their isolated compounds have been described in the literature. However, to our knowledge, there are no validated analytical methods for the analysis and standardization of products derived from this type of propolis reported in the literature. We developed a reverse-phase high-performance liquid chromatography analytical method for the detection and quantification of nine red propolis chemical markers: liquiritigenin, calycosin, isoliquiritigenin , formononetin, vestitol, neovestitol, medicarpin, biochanin A, and 7- O -methylvestitol, present in Brazilian red propolis extracts and D. ecastaphyllum. The developed method was also applied to the analyses of D. ecastaphyllum samples and seasonal analysis of Brazilian red propolis. Good detection response, linearity, precision, and robustness were obtained by the method, being reliable for the quality control of Brazilian red propolis extracts, raw propolis, plant material, and their derived products. The red propolis chemical markers were present in D. ecastaphyllum stems at lower concentrations. The seasonal analysis of Brazilian red propolis extract showed higher phenolic compound concentration on periods of the rainy season with higher humidity and lower solar radiation. [ABSTRACT FROM AUTHOR]

[Display omitted] The synthetic polyhexamethylene guanidine hydrochloride (PHMGH) polymer presents antifungal and antimicrobial activities in vitro. However, in vivo reports regarding its antiseptic and healing activity are scarce in the scientific literature. Thus, the present study aimed to evaluate the antimicrobial and healing effects, as well as toxicological parameters, of a topical solution containing 0.5% PHMGH (Akwaton®) in the treatment of superficial skin wounds experimentally induced on the dorsum of rodents. In addition, non-clinical safety studies were also conducted for use in human health, such as acute oral toxicity and genotoxicity tests. Animals did clinically not present dermatitis. After two days of topical treatment, PHMGH showed a significant antiseptic effect compared to the untreated group, reducing the number of colony-forming units by 72%, reaching 100% on the fourth day of treatment. The animals treated with PHMGH showed a significant area reduction of the skin lesions in relation to the untreated group, indicating a healing effect of the polymer. Moreover, PHMGH treatment led to a significant increase in fibroblasts when compared to the untreated group, revealing its healing action. No significant differences were observed between the biochemical indicators of hepatoxicity and nephrotoxicity, nor genotoxicity between the PHMGH-treated and the negative control groups. The results of acute oral toxicity showed that PHMGH at 5% presents a lethal dose 50% greater than the 2000 mg/kg. At a concentration of 5%, PHMGH did not show genotoxicity nor cytotoxicity at doses up to 1500 mg/kg through the micronucleus assay in mice. Therefore, 0.5% PHMGH showed an antimicrobial and healing effect, with no toxicity, and could be a promising adjunct in the microbial control of healing wounds. [ABSTRACT FROM AUTHOR]

• Dental caries and endodontic infections are the most common oral infectious diseases. • The C. pubiflora oleoresin and ent -hardwickiic acid display promising activity. • C. pubiflora oleoresin and ent -hardwiickic acid damaged the bacterial membrane. The microorganisms that constitute the oral microbiome can cause oral diseases, including dental caries and endodontic infections. The use of natural products could help to overcome bacterial resistance to the antimicrobials that are currently employed in clinical therapy. This study assessed the antimicrobial activity of the Copaifera pubiflora oleoresin and of the compounds isolated from this resin against oral bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays provided values ranging from 6.25 to > 400 μg/mL for the C. pubiflora oleoresin and its isolated compounds. The fractional inhibitory concentration index (FICI) assay showed that the oleoresin and chlorhexidine did not act synergistically. All the tested bacterial strains formed biofilms. MICB 50 determination revealed inhibitory action: values varied from 3.12–25 μg/mL for the oleoresin, and from 0.78 to 25 μg/mL for the ent -hardwickiic acid. Concerning biofilm eradication, the C. pubiflora oleoresin and hardwickiic acid eradicated 99.9 % of some bacterial biofilms. Acid resistance determination showed that S. mutans was resistant to acid in the presence of the oleoresin and ent -hardwickiic acid at pH 4.0, 4.5, and 5.0 at all the tested concentrations. Analysis of DNA/RNA and protein release by the cell membrane demonstrated that the oleoresin and hardwiickic acid damaged the bacterial membrane irreversibly, which affected membrane integrity. Therefore, the C. pubiflora oleoresin and ent -hardwickiic acid have potential antibacterial effect and can be used as new therapeutic alternatives to treat oral diseases such as dental caries and endodontic infections. [ABSTRACT FROM AUTHOR]

Being aware of the remarkable antimicrobial potential of S. officinalis L., we aimed to evaluate the antimicrobial activity of the S. officinalis dichloromethane crude extract (SOD), dichloromethane-soluble fractions (SODH and SODD), SODD subfractions (SODD1 and SODD2), and pure substances (manool, salvigenin, and viridiflorol) against periodontopathogens. This bioassay-guided study comprises five antimicrobial tests—determination of the Minimum Inhibitory Concentration (MIC), determination of the Minimum Bactericidal Concentration (MBC), determination of the antibiofilm activity, construction of the Time-kill curve (determination of Bactericidal Kinetics), and determination of the Fractional Inhibitory Concentration Index—on six clinical bacterial isolates and three standard bacterial strains involved in periodontal disease. SOD has moderate activity against most of the tested bacteria, whereas SODD1, SODH1, SODH3, and manool afford the lowest results. The Porphyromonas gingivalis (ATTC and clinical isolate) biofilm is considerably resistant to all the samples. In association with chlorhexidine gluconate, only SODH1 exerts additive action against P. gingivalis (clinical isolate). Therefore, SODH1 and manool are promising antibacterial agents and may provide therapeutic solutions for periodontal infections. • S. officinalis crude extract had moderate action against all the bacteria tested. • Subfractions the bioguided study (SODD1, SODH1, SODH3), and manool compound proved to be promising antimicrobial agents. • The secondary metabolites of S. officinalis may provide therapeutic solutions for periodontal infections. [ABSTRACT FROM AUTHOR]