Chemical composition and anti-inflammatory activity of the essential oils of Piper gaudichaudianum and Piper mikanianum.

ETHNOPHARMACOLOGICAL RELEVANCE: Inflammation is the body's normal protective response to injury and is stimulated by pathogens, toxic compounds, damaged cells or radiation, promoting healing and restoring homeostasis to the injured tissue. Leaves of Piper gaudichaudianum Kunth, known as "pariparoba" are widely used in folk medicine for the relief of toothache, while the fresh roots are used as anti-inflammatory and to treat liver disorders. For P. mikanianum (Kunth) Steud is known as "aguaxima", is widely used in the treatment of inflammation, rheumatism and ulcer, with its roots being used for stomach disorders and as a diaphoretic in intermittent fevers. AIM OF THE STUDY: Therefore, this work aims to chemically characterize the essential oil of Piper gaudichaudianum and Piper mikanianum, as well as the evaluation of neutrophil antichemotactic activity of both essential oils in order to complement the information of its traditional use, taking the leaves as plant material and, with that, corroborating its use in folk medicine for the treatment of inflammatory diseases. MATERIALS AND METHODS: The essential oil from leaves of both Piper species were obtained from crushed fresh samples, by hydrodistillation using a Clevenger type-apparatus for 4 h. The yield determination was performed as vol/wt (v/w) and in triplicate. The amount of essential oil obtained was quantified in mL. The identification and quantification of the compounds was performed using gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The in vitro anti-inflammatory activity was evaluated using the model of modified Boyden chamber. In this test the essential oils were tested for their ability to inhibit leukocyte chemotaxis stimulated by Escherichia coli lipopolysaccharide. RESULTS: The chemical composition of the essential oils revealed the identification of 26 constituents for P. gaudichaudianum being the sesquiterpenes ß-selinene (14.0%) and viridiflorene (10.5%) the main compounds, followed by caryophyllene oxide (9.3%) and (E)-nerolidol (9.0%). For P. mikanianum essential oil, ß-myrcene (17.2%) and bicyclogermacrene (26.3%) were the major components in the monoterpenes and sesquiterpene fractions, respectively. The essential oils were also tested for their ability to inhibit neutrophil chemotaxis in vitro when stimulated by Escherichia coli lipopolysaccharide. Both essential oils showed antichemotactic effect with reduction in migration of 0-72.2% for P. gaudichaudianum and 8.6-100% for P. mikanianum to same concentrations, suggesting a response to acute inflammatory processes. CONCLUSIONS: Since up to date there is no report of this biological activities by this mechanism (antichemotactic assay) for essential oils this species. These results showed that the essential oils of P. gaudichaudianum and P. mikanianum have a great capacity to inhibit neutrophil chemotaxis in an inflammatory process, in a dose-dependent way, suggesting anti-inflammatory potential, by preventing its accumulation at the injury site with the possibility of tissue damage. Findings of these studies support the traditional use of these species in the treatment of inflammatory processes.

Polyphenolic Composition and in Vitro Antihypertensive and Anti-Inflammatory Effects of Cuphea lindmaniana and Cuphea urbaniana.

The present study investigates the chemical composition, anti-inflammatory, and antihypertensive activities, in vitro, from extracts of Cuphea lindmaniana and Cuphea urbaniana leaves. The extraction was performed ultrasound-assisted, and UHPLC/MS analysis was in positive mode ionization. The anti-inflammatory activity of the extracts and miquelianin were assayed at concentrations 0.001-10 µg/mL by chemotaxis on rat polymorphonuclear neutrophils. The antihypertensive activity was performed by angiotensin-converting enzyme (ACE) inhibition. From the nineteen proposed compounds, six of them are described for the first time in this genus. The extracts displayed antichemotactic effect with a reduction of 100 % of the neutrophil migration, in vitro, in most concentrations. The ACE-inhibition presented results ranging from 19.58 to 22.82 %. In conclusion, C. lindmaniana and C. urbaniana extracts contain a rich diversity of flavonoids and display in vitro anti-inflammatory and antihypertensive potential. Thus, this study could serve as a scientific baseline for further investigation, on developmental novel products with therapeutic actions.

Cuphea spp.: antichemotactic study for a potential anti-inflammatory drug.

Cuphea genus (Lythraceae) popularly known in Brazil as "sete-sangrias", it's described as antimicrobial, anti-inflammatory, diuretic and antihypertensive mainly. Investigating the chemotactic ability plays an important role in the identification of new anti-inflammatory agents. Thus, this research aims to assay the antichemotactic activity of hydroethanolic extracts of C. calophylla, C. carthagenensis, C. glutinosa, and C. racemosa as well as the compounds miquelianin and myricitrin. The antichemotactic activity of the hydroethanolic extracts, miquelianin, and myricitrin were assayed at concentrations 0.001 to 10 µg/mL in the lipopolysaccharide-induced chemotaxis on rat polymorphonuclear neutrophils. All the assayed samples displayed antichemotactic activity with reduction of the neutrophil migration in the range of 4.46-100%, and an IC50 value in the range of 0.30-1.24 µg/mL. Thus, this study demonstrates that the extracts hydroethanolic of Cuphea species, miquelianin, and myricitrin display a significant antichemotactic activity. Therefore, in future studies, extracts from Cuphea spp. could be used as anti-inflammatory drugs.

Polyphenols composition from leaves of Cuphea spp. and inhibitor potential, in vitro, of angiotensin I-converting enzyme (ACE).

ETHNOPHARMACOLOGICAL RELEVANCE: Cuphea is the largest genus of the Lythraceae family. It is popularly known as "sete-sangrias" in Brazil used in folk medicine as a diuretic, antipyretic, anti-inflammatory, laxative and antihypertensive agent. The raw material of Cuphea has shown promising results in the production of fitotherapics, which are chemically characterized by quercetin core flavonoids. AIMS OF THE STUDY: Present work aims to investigate the chemical composition of Cuphea calophylla, Cuphea carthagenensis, Cuphea glutinosa and Cuphea racemosa by UHPLC-MS using ESI-Q-TOF, and also to investigate the inhibition of angiotensin-converting enzyme (ACE) in vitro. MATERIALS AND METHODS: Leaves extraction was conducted by an ultrasound-assisted system under the following conditions: 40% ethanol, particle size ≤180 µm, plant:solvent ratio 1:20 (w/v) for 30 min. The leaf extracts were analyzed by UHPLC-MS positive mode ionization. For the inhibition of ACE, the leaf extracts used were obtained from different Cuphea species collected from several regions of Rio Grande do Sul (Brazil). RESULTS: In total 26 polyphenolic compounds were proposed, which were mostly derived from quercetin, myricetin, and kaempferol. Of these compounds, ten are described in the genus for the first time. The ACE-inhibiting activities are presented in descending order: miquelianin (32.41%), C. glutinosa 1 (31.66%), C. glutinosa 5 (26.32%) and C. carthagenensis 1 (26.12%). CONCLUSION: The obtained results suggest that the ACE-inhibiting potential may be increased by the interactions among the different phytoconstituents present in the crude extract. These results corroborate with the popular usage of Cuphea genus as diuretic and antihypertensive agents in folk medicine.

Chemical Constituents and Pharmacology properties of Aristolochia triangularis: a south brazilian highly-consumed botanical with multiple bioactivities.

Aristolochia triangularis Cham., is one of the most frequently used medicinal plant in Southern Brazil. Preparations containing the leaves and/or stems are traditionally used as anti-inflammatory, diuretic, as well as antidote against snakebites. This study screened A. triangularis extracts, fractions and isolated compounds for different bioactivities. A weak antiproliferative activity against human lung cancer cell line (A549) was observed only for chloroform fraction obtained from stems (CFstems - CC50: 2.93 µg/mL). Also, a moderate antimicrobial activity against Staphylococcus aureus was detected just for chloroform fraction obtained from leaves (CFleaves -13-16 mm inhibition zone). Additionally, two semi-purified fractions (CFstems-4 and CFleaves-4) selectively inhibited HSV-1 replication (IC50 values of 0.40 and 2.61 µg/mL, respectively), while only CFleaves showed promising results against Leishmania amazonensis. Fractionation of extracts resulted in the isolation of one neolignan (-) cubebin and one lignan (+) galbacin. However, these compounds are not responsible for the in vitro bioactivities herein detected. The presence of aristolochic acid I and aristolochic acid II in the crude ethanol extract of stems (CEEstems) and leaves (CEEleaves) was also investigated. The HPLC analysis of these extracts did not display any peak with retention time or UV spectra comparable to aristolochic acids I and II.

Antichemotactic and Antifungal Action of the Essential Oils from Cryptocarya aschersoniana, Schinus terebinthifolia, and Cinnamomum amoenum.

The purpose of this work was to determine the chemical composition and evaluate the antichemotactic, antioxidant, and antifungal activities of the essential oil obtained from the species Cryptocarya aschersoniana Mez, Cinnamomum amoenum (Ness & Mart.) Kosterm., and Schinus terebinthifolia Raddi, as well as the combination of C. aschersoniana essential oil and terbinafine against isolates of dermatophytes. Allo-aromadendrene, bicyclogermacrene, and germacrene B were identified as major compounds in essential oils. The essential oil of C. aschersoniana shown 100 % inhibitory effect on leukocyte migration at the concentration of 10 µg/mL while S. terebinthifolia oil presented 80.1 % inhibitory effect at the same concentration. Only S. terebinthifolia oil possessed free-radical-scavenging activity which indicates its antioxidant capacity. The essential oils were also tested against fungal isolates of dermatophyte species (Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis and Microsporum gypseum), resulting in MIC ranging from 125 µg/mL to over 500 µg/mL. C. aschersoniana oil combined with terbinafine resulted in an additive interaction effect. In this case, the essential oil may act as a complement to conventional therapy for the topical treatment of superficial fungal infections, mainly because it is associated with an anti-inflammatory effect.

Leishmanicidal and antichemotactic activities of icetexanes from Salvia uliginosa Benth.

BACKGROUND: Several species of Salvia are used as medicinal plants around the world. Biological activities of isolated compounds have been described, being diterpenes frequently responsible for the effects. PURPOSE: Isolation of diterpenes from Salvia uliginosa Benth. and evaluation of the antichemotactic and leishmanicidal activities of the isolated compounds. STUDY DESIGN: To isolate diterpenes from S. uliginosa and evaluate their antichemotactic and leishmanicidal activities in vitro. METHODS: The exudate of S. uliginosa was obtained by rapidly dipping the aerial parts in dichloromethane. The compounds were isolated by repeated column chromatography over silica gel. The effects on L. amazonensis growth, survival, DNA degradation, ROS generation, as well as the antichemotactic activity and cytotoxicity of the compounds towards human erythrocytes and macrophages were evaluated. RESULTS: A novel icetexane diterpene, isoicetexone (IsoICT) along with the known diterpenes icetexone (ICT), and 7-acetoxy-6,7-dihydroicetexone were isolated from the dichloromethane surface exudate of S. uliginosa. The structures were elucidated using NMR and MS experiments, and by comparison with previously reported data. IsoICT and ICT at low concentrations caused completely inhibition of neutrophils migration in vitro. In addition, IsoICT and ICT showed high leishmanicidal activity against L. amazonensis, induced ROS production in parasites and presented low cytotoxicity against macrophages and human erythrocytes, and moderate to high selectivity index. CONCLUSION: These data indicated that IsoICT and ICT exhibit potent antichemotactic and leishmanicidal effects. Further studies are needed in order to evaluate the in vivo activities as well as the toxicity of the compounds.

Toxicological profile and acetylcholinesterase inhibitory potential of Palicourea deflexa, a source of ß-carboline alkaloids.

Palicourea genus is chemically and taxonomically close to Psychotria genus, a well-known source of neuroactive alkaloids. It has been previously reported the pharmacological potential of these alkaloids in some targets related to the neurodegenerative process. In this context, this study was carried out in order to evaluate the toxic effects and acetylcholinesterase (AChE) inhibitory potential of Palicourea deflexa fraction of total alkaloids (FTA). P. deflexa FTA was analyzed by means of HPLC-DAD and HRMS-ESI. We performed toxicological screening through Fish Embryo Toxicity (FET) test using zebrafish embryo and abnormal developmental phenotypes were recorded daily. For AChE inhibition, zebrafish brains were used as enzymatic source and formation of thiolate dianion of 5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB) was used to monitor acetylthiocholine hydrolysis. Lineaweaver-Burk double reciprocal plots were used to indicate mode of inhibition. Chemical analysis of the P. deflexa FTA allowed the identification of the main compound as harman-3-carboxylic acid. This fraction was evaluated in vivo for its toxicological effect. The zebrafish embryo test indicated that the FTA has a lethal concentration of 50% (LC50)=72.18µg/mL. Further, the FTA was evaluated for its AChE inhibitory profile, demonstrating an inhibitory concentration of 50% (IC50) of 50.65µg/mL. Lineaweaver-Burk double reciprocal plots indicated a mixed mode of inhibition. It is reported for the first time the toxicological and pharmacological profile of the alkaloid fraction of Palicourea deflexa in zebrafish models.

Dimeric acylphloroglucinols from Hypericum austrobrasiliense exhibiting antinociceptive activity in mice.

Three dimeric acylphloroglucinols, austrobrasilol A, austrobrasilol B and isoaustrobrasilol B were isolated from the flowers of Hypericum austrobrasiliense (Hypericaceae, section Trigynobrathys). Their structures were elucidated using mass spectrometry and NMR experiments (1D and 2D), and by comparison with previously reported data for other dimeric acylphloroglucinols isolated from Hypericum and Elaphoglossum genera. The three compounds were orally administered in mice at equimolar doses to uliginosin B (15mg/kg, p.o.) displaying antinociceptive activity in the hot-plate test. The compounds did not induce motor impairment in the rotarod apparatus.

Chemical analysis and in vitro antiviral and antifungal activities of essential oils from Glechon spathulata and Glechon marifolia.

CONTEXT: Glechon spathulata Benth. and Glechon marifolia Benth. (Lamiaceae, Mentheae) are aromatic plants used in traditional medicine for the treatment of viral infections. OBJECTIVE: The chemical composition and antiviral and antifungal activities of Glechon spathulata and Glechon marifolia essential oils were investigated. MATERIALS AND METHODS: The oils were obtained by hydrodistillation and analyzed by GC-FID and GC-MS. Anti-herpes virus (HSV-1) activity was examined in Vero cells by yield reduction assay, in doses of 0.0095% v/v and 0.039% v/v, for G. spathulata and G. marifolia oil, respectively. Antifungal activity was carried by the broth microdilution method, in oil concentrations that ranged from 5.2 to 500 µg/mL. RESULTS: ß-Caryophyllene (14.2% and 32.2% for G. spathulata and G. marifolia, respectively) and bicyclogermacrene (17.1% and 16.5%, respectively) were the major components of both oils. At noncytotoxic concentrations of the essential oils, the viral titer was reduced by up to 2 log10 for KOS and VR-733 strains. The antifungal activity was observed against Trichophyton rubrum (MIC 10-83 µg/mL) and Epidermophyton floccosum (MIC 83-500 µg/mL). The oil of G. spathulata exhibited activity against the three strains tested (KOS, VR733, and 29-R), whereas G. marifolia oil was active against two strains, KOS and VR733. DISCUSSION AND CONCLUSION: The chemical composition for G. spathulata and G. marifolia essential oils is very similar. The oil of G. spathulata can be promising as a new antifungal agent against dermatophytes. The findings add important information to the biological activity of Glechon species essential oils, specifically its antiviral and antidermatophytic properties.

Palynological analysis of a late Holocene core from Santo Antônio da Patrulha, Rio Grande do Sul, Southern Brazil

A sedimentar core collected at Santo Antônio da Patrulha, Rio Grande do Sul State, southmost Brazil, was submitted to pollen analysis to provide the vegetational history of this region, and the paleoecological and paleoclimatic changes. A total of 98 taxa of palynomorphs was identified from 35 subsamples. Three radiocarbonic datings were obtained along a section of 115 cm depth, including the basal age of 4730 ± 50 yr BP. Pollen diagrams and cluster analysis were performed based on palynomorphs frequencies, demonstrating five distinct phases (SAP-I to SAP-V), which reflected different paleoecological conditions. The predominance of plants associated with grasslands in the phase SAP-I suggests warm and dry climate conditions. A gradual increasing of humidity conditions was observed mainly from the beginning of the phase SAP-III, when the vegetation set a mosaic of grasslands and Atlantic rainforest. Furthermore, the presence of some forest taxa ( Acacia-type, Daphnopsis racemosa, Erythrina-type and Parapiptadenia rigida-type), from the phase SAP-IV, is interpreted as an influence of the seasonal semideciduous forest in the study region. From the phase SAP-V (ca. 4000 yrs BP), the vegetation became similar to the modern one (extant Atlantic rainforest Biome), especially after 2000 yrs BP (calibrated age).

Um testemunho de sondagem coletado em Santo Antônio da Patrulha, Rio Grande do Sul, Brasil, foi submetido para análise polínica a fim de revelar a história vegetacional e mudanças paleoecológicas e paleoclimáticas. Um total de 98 táxons foi identificado a partir de 35 subamostras. Três datações radiocarbônicas foram obtidas ao longo de uma seção de 115 cm de profundidade, incluindo a idade basal de 4730 ± 50 anos AP. Diagramas polínicos e análises de agrupamentos foramrealizadas com base nas freqüências dos palinomorfos, demonstrando cinco fases distintas (SAP-I a SAP-V), as quais refletiram diferentes condições paleoecológicas. A predominância de plantas relacionadas à vegetação campestre na fase SAP-I sugere condições climáticas quentes e secas. Um gradual aumento nas condições de umidade foi observado principalmente no início da fase SAP-III, quando a vegetação conformou um mosaico de Campos e Floresta Atlântica. Além disso, a presença de certos táxons florestais (tipo- Acacia, Daphnopsis racemosa, tipo Erythrina e tipo Parapiptadenia rigida), a partir da fase SAP-IV, é interpretada como influência da Floresta Estacional Semidecidual na região de estudo. A partir da fase SAP-V (ca. 4000 anos AP) a vegetação tornou-se similar à moderna (atual Bioma da Floresta Atlântica), especialmente após 2000 anos AP (idade calibrada).

Palynological analysis of a late Holocene core from Santo Antônio da Patrulha, Rio Grande do Sul, Southern Brazil.

A sedimentar core collected at Santo Antônio da Patrulha, Rio Grande do Sul State, southmost Brazil, was submitted to pollen analysis to provide the vegetational history of this region, and the paleoecological and paleoclimatic changes. A total of 98 taxa of palynomorphs was identified from 35 subsamples. Three radiocarbonic datings were obtained along a section of 115 cm depth, including the basal age of 4730 ± 50 yr BP. Pollen diagrams and cluster analysis were performed based on palynomorphs frequencies, demonstrating five distinct phases (SAP-I to SAP-V), which reflected different paleoecological conditions. The predominance of plants associated with grasslands in the phase SAP-I suggests warm and dry climate conditions. A gradual increasing of humidity conditions was observed mainly from the beginning of the phase SAP-III, when the vegetation set a mosaic of grasslands and Atlantic rainforest. Furthermore, the presence of some forest taxa ( Acacia-type, Daphnopsis racemosa, Erythrina-type and Parapiptadenia rigida-type), from the phase SAP-IV, is interpreted as an influence of the seasonal semideciduous forest in the study region. From the phase SAP-V (ca. 4000 yrs BP), the vegetation became similar to the modern one (extant Atlantic rainforest Biome), especially after 2000 yrs BP (calibrated age).

Acaricidal properties of extracts from the aerial parts of Hypericum polyanthemum on the cattle tick Boophilus microplus.

Laboratory tests were carried out on larvae and adults of the cattle tick Boophilus microplus to determine the toxicity of n-hexane and crude methanolic extracts of Hypericum polyanthemum (Guttiferae) using the larval immersion test (LIT) and adult immersion test (AIT). In the AIT, the effectiveness of treatment against engorged females was assessed by measuring egg production: the n-hexane extract was found to have a small effect on the egg laying at the highest concentration (19.2% of egg-laying inhibition) whilst the crude methanolic extract did not affect the egg production. For the LIT the n-hexane extracts were highly toxic to the larvae at all the concentrations (100% of mortality). The crude methanolic extract was also toxic to the larvae at higher concentrations killing 100, 96.7, 84.7 and 52.7% at the concentrations of 50, 25, 12.5, and 6.25 mg/ml, respectively, 48 h after the immersion of the acarus.

Benzophenones from Hypericum carinatum.

Two new benzophenones were isolated from the leaves of Hypericum carinatum. Their structures were established on the basis of 2D NMR spectroscopic analyses and mass spectrometry as cariphenone A (6-benzoyl-5,7-dihydroxy-2,2,8-trimethyl-2H-chromene) (1) and cariphenone B (8-benzoyl-5,7-dihydroxy-2,2,6-trimethyl-2H-chromene) (2). Five known compounds, the phloroglucinol derivative uliginosin B (3), 1-eicosanol, sitosterol, stigmasterol, and campesterol, were also characterized. Compounds 1-3 were evaluated for their total antioxidant capacity through a total radical-trapping parameter assay. Only compound 1 showed moderate antioxidant activity, exhibiting inhibition of chemiluminescence similar to that of quercetin at the same concentration.

Screening of Brazilian plants for the presence of peroxides

Chloroform or dichloromethane extracts of 357 southern Brazilian plant species were tested for the presence of peroxides by thin-layer chromatography, using the spray reagent from Huber & Fröhlke. >From the species tested, 71 (20 por cento) showed positive results and most of them (56 por cento) are Asteraceae species. The species tested were mainly from Asteraceae, but 55 more families were screened, in a total of 77 genera surveyed...