Annexin A1 Mimetic Peptide and Piperlongumine: Anti-Inflammatory Profiles in Endotoxin-Induced Uveitis.

Uveitis is one of the main causes of blindness worldwide, and therapeutic alternatives are worthy of study. We investigated the effects of piperlongumine (PL) and/or annexin A1 (AnxA1) mimetic peptide Ac2-26 on endotoxin-induced uveitis (EIU). Rats were inoculated with lipopolysaccharide (LPS) and intraperitoneally treated with Ac2-26 (200 µg), PL (200 and 400 µg), or Ac2-26 + PL after 15 min. Then, 24 h after LPS inoculation, leukocytes in aqueous humor, mononuclear cells, AnxA1, formyl peptide receptor (fpr)1, fpr2, and cyclooxygenase (COX)-2 were evaluated in the ocular tissues, along with inflammatory mediators in the blood and macerated supernatant. Decreased leukocyte influx, levels of inflammatory mediators, and COX-2 expression confirmed the anti-inflammatory actions of the peptide and pointed to the protective effects of PL at higher dosage. However, when PL and Ac2-26 were administered in combination, the inflammatory potential was lost. AnxA1 expression was elevated among groups treated with PL or Ac2-26 + PL but reduced after treatment with Ac2-26. Fpr2 expression was increased only in untreated EIU and Ac2-26 groups. The interaction between Ac2-26 and PL negatively affected the anti-inflammatory action of Ac2-26 or PL. We emphasize that the anti-inflammatory effects of PL can be used as a therapeutic strategy to protect against uveitis.

Pharmacological and Therapeutic Potential of Myristicin: A Literature Review.

Natural products have been used by humanity for many centuries to treat various illnesses and with the advancement of technology, it became possible to isolate the substances responsible for the beneficial effects of these products, as well as to understand their mechanisms. In this context, myristicin, a substance of natural origin, has shown several promising activities in a large number of in vitro and in vivo studies carried out. This molecule is found in plants such as nutmeg, parsley, carrots, peppers, and several species endemic to the Asian continent. The purpose of this review article is to discuss data published in the last 10 years at Pubmed, Lilacs and Scielo databases, reporting beneficial effects, toxicity and promising data of myristicin for its future use in medicine. From 94 articles found in the literature, 68 were included. Exclusion criteria took into account articles whose tested extracts did not have myristicin as one of the major compounds.

mRNAs biomarker related to the control of proliferation and cell death in HepG2/C3A spheroid and monolayer cultures treated with piperlongumine

Background: Cell culture (spheroid and 2D monolayer cultures) is an essential tool in drug discovery. Piperlongumine (PLN), a naturally occurring alkaloid present in the long pepper (Piper longum), has been implicated in the regulation of GSTP1 activity. In vitro treatment of cancer cells with PLN increases ROS (reactive oxygen species) levels and induces cell death, but its molecular mode of action has not been entirely elucidated. Methods: In this study, we correlated the antiproliferative effects (2D and 3D cultures) of PLN (CAS 20069­09-4, Sigma-Aldrich) with morphological and molecular analyses in HepG2/C3A cell line. We performed assays for cytotoxicity (MTT), comet assays for genotoxicity, induction of apoptosis, analysis of the cell cycle phase, and analysis of the membrane integrity by flow cytometry. Relative expression of mRNA of genes related to proliferation, apoptosis, cell cycle control, metabolism of xenobiotics, and reticulum endoplasmic stress. Results: PLN reduced the cell proliferation by the cell cycle arrest in G2/M. Changes in the mRNA expression for CDKN1A (4.9x) and CCNA2 (0.5x) of cell cycle control genes were observed. Cell death occurred due to apoptosis, which may have been induced by increased expression of proapoptotic mRNAs (BAK1, 3.1x; BBC3, 2.4x), and by an increase in 9 and 3/7 active caspases. PLN induced cellular injury by ROS generation and DNA damage. DNA damage induced MDM2 signaling (3.0x) associated with the appearance of the monastral spindle in mitosis. Genes associated with ROS degradation also showed increased mRNA expression (GSR, 2.0x; SOD1, 2.1x). PLN induce endoplasmic reticulum stress with the increase in the mRNA expression of ERN1 (4.5x) and HSPA14 (2.2x). The xenobiotic metabolism showed increased mRNA expression for CYP1A2 (2.2x) and CYP3A4 (3.4x). In addition to 2D culture, PLN treatment also inhibited the growth of 3D culture (spheroids). Conclusion: Thus, the findings of our study show that several gene expression biomarkers (mRNAs) and monastral spindle formation indicated the many pathways of damage induced by PLN treatment that contributes to its antiproliferative effects

A screening of the MMV Pathogen Box® reveals new potential antifungal drugs against the etiologic agents of chromoblastomycosis.

Chromoblastomycosis (CBM) is a chronic subcutaneous mycosis caused by traumatic implantation of many species of black fungi. Due to the refractoriness of some cases and common recurrence of CBM, a more effective and less time-consuming treatment is mandatory. The aim of this study was to identify compounds with in vitro antifungal activity in the Pathogen Box® compound collection against different CBM agents. Synergism of these compounds with drugs currently used to treat CBM was also assessed. An initial screening of the drugs present in this collection at 1 µM was performed with a Fonsecaea pedrosoi clinical strain according to the EUCAST protocol. The compounds with activity against this fungus were also tested against other seven etiologic agents of CBM (Cladophialophora carrionii, Phialophora verrucosa, Exophiala jeanselmei, Exophiala dermatitidis, Fonsecaea monophora, Fonsecaea nubica, and Rhinocladiella similis) at concentrations ranging from 0.039 to 10 µM. The analysis of potential synergism of these compounds with itraconazole and terbinafine was performed by the checkerboard method. Eight compounds inhibited more than 60% of the F. pedrosoi growth: difenoconazole, bitertanol, iodoquinol, azoxystrobin, MMV688179, MMV021013, trifloxystrobin, and auranofin. Iodoquinol produced the lowest MIC values (1.25-2.5 µM) and MMV688179 showed MICs that were higher than all compounds tested (5 - >10 µM). When auranofin and itraconazole were tested in combination, a synergistic interaction (FICI = 0.37) was observed against the C. carrionii isolate. Toxicity analysis revealed that MMV021013 showed high selectivity indices (SI ≥ 10) against the fungi tested. In summary, auranofin, iodoquinol, and MMV021013 were identified as promising compounds to be tested in CBM models of infection.

Effect of piperlongumine during exposure to cigarette smoke reduces inflammation and lung injury.

Chronic obstructive pulmonary disease (COPD) is related to smoking and anti-inflammatory therapy is indicated. Among the mediators with anti-inflammatory properties, we highlight piperlongumine (PL), an alkaloid/amide of Piper longum. Here we evaluated the PL administration on an experimental model of respiratory inflammation resulting from exposure to cigarette smoke. Male Balb/c mice were exposed to burning of 10 commercial cigarettes, 2x/day, for five weeks on specific equipment. PL efficacy was evaluated in control, exposed to smoke without treatment and PL treated (2.0 mg/kg, 3x/week) groups. Animals were weighed and plethysmographic analyses performed at the end of the exposure protocol. Inflammatory cells were evaluated in the bronchoalveolar lavage (BAL) and hemoglobin and glucose in the blood. Lung fragments were processed for histopathological studies and AnxA1, COX-2, NF-kB and neutrophil elastase expressions. Plethysmography revealed that PL maintained pulmonary frequency, volume and ventilation parameters similar to controls, with respiratory volume reduction compared to untreated animals. Final weight was reduced in both exposed groups. PL decreased hemoglobin concentration, attenuated the reduction of glucose levels and reduced influx of lymphocytes, neutrophils and macrophages in BAL. Histopathologically occured infiltration of inflammatory cells, increase of the interalveolar septa and intra-alveolar spaces in untreated animals. But, PL administration recovered lung tissues and, immunohistochemically, promoted increased expression of AnxA1 and reduction of COX-2, NF-kB and neutrophil elastase. Together the results indicate that PL attenuates systemic and pulmonary inflammatory changes, partially by modulating the expression the endogenous AnxA1, and may represent a promising therapy in preventing the inflammation induced by cigarette smoke.

Immune Modulation by Inhibitors of the HO System.

The heme oxygenase (HO) system involves three isoforms of this enzyme, HO-1, HO-2, and HO-3. The three of them display the same catalytic activity, oxidating the heme group to produce biliverdin, ferrous iron, and carbon monoxide (CO). HO-1 is the isoform most widely studied in proinflammatory diseases because treatments that overexpress this enzyme promote the generation of anti-inflammatory products. However, neonatal jaundice (hyperbilirubinemia) derived from HO overexpression led to the development of inhibitors, such as those based on metaloproto- and meso-porphyrins inhibitors with competitive activity. Further, non-competitive inhibitors have also been identified, such as synthetic and natural imidazole-dioxolane-based, small synthetic molecules, inhibitors of the enzyme regulation pathway, and genetic engineering using iRNA or CRISPR cas9. Despite most of the applications of the HO inhibitors being related to metabolic diseases, the beneficial effects of these molecules in immune-mediated diseases have also emerged. Different medical implications, including cancer, Alzheimer´s disease, and infections, are discussed in this article and as to how the selective inhibition of HO isoforms may contribute to the treatment of these ailments.

In vitro anthelmintic activity of the crude hydroalcoholic extract of Piper cubeba fruits and isolated natural products against gastrointestinal nematodes in sheep.

This study describes the in vitro anthelmintic activity of a hydroalcoholic extract from the fruit of Piper cubeba and its major isolated components against the eggs and larvae of gastrointestinal nematodes obtained from naturally-infected ovines. In vitro anthelmintic activity was evaluated using the egg hatch test (EHT), larval development test (LDT) and L3 migration inhibition test (LMT). The extract showed ovicidal and larvicidal activity, with an EC50 of 200 µg/mL and 83.00 µg/mL in the EHT and LDT, respectively. The extract inhibited 100% of larval migration at the lowest tested concentration (95 µg/mL). The crude extract was purified using successive silica gel chromatographic columns, which revealed the lignans hinokinin, cubebin and dihydrocubebin as the major compounds that were present, which were then used in in vitro tests. Cubebin, dihydrocubebin and hinokinin showed higher activity than the crude extract, with an EC50 for ovicidal activity of 150.00 µg/mL, 186.70 µg/mL and 68.38 µg/mL, respectively. In the LDT, cubebin presented an EC50 of 14.89 µg/mL and dihydrocubebin of 30.75 µg/mL. Hinokinin inhibited 100% the larval development at all concentrations evaluated. In the LMT, dihydrocubebin inhibited 100% the larval migration in all concentrations evaluated while cubebin and hinokinin showed EC50 values of 0.89 µg/mL and 0.34 µg/mL, respectively. P. cubeba extract is rich in several classes of active compounds, but here we demonstrate that the described anthelmintic activity may be related to the presence of these lignans, which are present in larger concentrations than other components of the extract. Our results demonstrate for first time the anthelmintic activity against gastrointestinal nematodes in sheep for this class of special metabolites that are present in P. cubeba fruit. However, future detailed studies are needed to evaluate the effectiveness of P. cubeba fruits extract and active lignans in in vivo tests.

Piperlongumine Induces Apoptosis in Colorectal Cancer HCT 116 Cells Independent of Bax, p21 and p53 Status.

BACKGROUND/AIM: Colorectal cancer is a common type of cancer with reported resistance to treatment, in most cases due to loss of function of apoptotic and cell-cycle proteins. Piperlongumine (PPLGM) is a natural alkaloid isolated from Piper species, with promising anti-cancer properties. This study investigated whether PPLGM is able to induce cell death in colorectal carcinoma HCT 116 cells expressing wild-type or deficient in Bax, p21 or p53. MATERIALS AND METHODS: PPLGM was extracted from roots of Piper tuberculatum. Cell viability was determined by reduction of 3-(4,5-dimethilthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and clonogenic assay. Cell death was evaluated by acridine orange/ethidium bromide staining and flow cytometry. Plasmid cleavage activity and circular dichroism DNA interaction were also analyzed. RESULTS: PPLGM induced selective cell death in all cell lines (IC50 range from 10.7 to 13.9 µM) with an increase in the number of late apoptotic cells and different profiles in cell-cycle distribution. Plasmid DNA analysis showed that PPLGM does not interact directly with DNA. CONCLUSION: This paper suggests that PPLGM may be a promising candidate in colorectal cancer therapy.

Leishmanicidal Effects of Piperlongumine (Piplartine) and Its Putative Metabolites.

Piperlongumine is an amide alkaloid found in Piperaceae species that shows a broad spectrum of biological properties, including antitumor and antiparasitic activities. Herein, the leishmanicidal effect of piperlongumine and its derivatives produced by a biomimetic model using metalloporphyrins was investigated. The results showed that IC50 values of piperlongumine in promastigote forms of Leishmania infantum and Leishmania amazonensis were 7.9 and 3.3 µM, respectively. The IC50 value of piperlongumine in the intracellular amastigote form of L. amazonensis was 0.4 µM, with a selectivity index of 25. The piperlongumine biomimetic derivatives, Ma and Mb, also showed leishmanicidal effects. We also carried out an in vitro metabolic degradation study showing that Ma is the most stable piperlongumine derivative in rat liver microsome incubations. The results presented here indicate that piperlongumine is a potential leishmanicidal candidate and support the biomimetic approach for development of new antileishmanial derivatives.

Effect of the Parafunctional Occlusal Loading and Crown Height on Stress Distribution

The aim of this study was to assess, by the three-dimensional finite element method, the influence of crown-to-implant ratio and parafunctional occlusal loading on stress distribution in single external hexagon implant-supported prosthesis. Computer-aided design software was used to confection three models. Each model was composed of a block bone and an external hexagon implant (5x10.0 mm) with screw-retained implant prostheses, varying the height crown: 10, 12.5 and 15 mm. Finite element analysis software was used to generate the finite element mesh and to establish the loading and boundary conditions. Normal (200 N axial and 100 N oblique load) and parafunctional forces (1,000 N axial and 500 N oblique load) were applied. The results were visualized by von Mises and maximum principal stress. In comparison with the normal occlusal force, the parafunctional occlusal force induced an increase in stress concentration and magnitude on implant (platform and first threads) and screw (neck). The cortical bone showed the highest tensile stress under parafunctional force (oblique load). The stress concentration increased as the crown height increased. It was concluded that: increasing the C/I increased stress concentration in both implant components and cortical bone; parafunctional loading increased between 4-5 times the value of stresses in bone tissue compared with functional loading; the type of loading variation factor is more influential than the crown-to-implant factor.

O objetivo deste estudo foi avaliar, através do método dos elementos finitos tridimensionais, a influência do carregamento oclusal parafuncional e da altura da coroa na distribuição das tensões em próteses unitárias implantossuportadas de hexágono externo. Foram confeccionados três modelos com o auxílio de programas de desenho assistido. Cada modelo foi composto por um bloco ósseo da região molar mandibular, por um implante de tipo hexágono externo (5x10,0 mm) e por coroa com diferentes alturas: 10, 12,5 e 15 mm. Os modelos foram exportados para o programa de elementos finitos NEiNastran 9.0, para geração das malhas e estabelecer as condições de contorno. Aplicou-se uma carga funcional (200 N axial e 100 N oblíqua), bem como uma carga parafuncional (1.000 N axial e 500 N oblíqua). Os resultados foram visualizados por meio de mapas de Tensão de von Mises e mapas de Tensão Máxima Principal. O carregamento parafuncional induziu um aumento da área de distribuição e da magnitude das tensões no implante (plataforma e primeiras roscas) e parafuso (pescoço) em comparação com o carregamento funcional. A cortical óssea apresentou maiores áreas de tensão por tração sob carregamento parafuncional oblíquo. A concentração de tensões aumentou à medida que aumentou a altura da coroa. O aumento da altura da coroa induziu um aumento na concentração de tensões, tanto nos componentes do implante, quanto na cortical óssea; o carregamento parafuncional induziu um aumento entre 4-5 vezes da magnitude das tensões no tecido ósseo; o tipo de carregamento apresenta-se como um fator de variação mais influente do que a proporção coroa/implante.

Piperlongumine inhibits NF-κB activity and attenuates aggressive growth characteristics of prostate cancer cells.

BACKGROUND: Elevated NF-κB activity has been previously demonstrated in prostate cancer cell lines as hormone-independent or metastatic characteristics develop. We look at the effects of piperlongumine (PL), a biologically active alkaloid/amide present in piper longum plant, on the NF-κB pathway in androgen-independent prostate cancer cells. METHODS: NF-κB activity was evaluated using Luciferase reporter assays and Western blot analysis of p50 and p65 nuclear translocation. IL-6, IL-8, and MMP-9 levels were assessed using ELISA. Cellular adhesion and invasiveness properties of prostate cancer cells treated with PL were also assessed. RESULTS: NF-κB DNA-binding activity was directly down-regulated with increasing concentrations of PL, along with decreased nuclear translocation of p50 and p65 subunits. Expression of IL-6, IL-8, MMP-9, and ICAM-1 was attenuated, and a decrease of cell-to-matrix adhesion and invasiveness properties of prostate cancer cells were observed. CONCLUSIONS: PL-mediated inhibition of NF-κB activity decreases aggressive growth characteristics of prostate cancer cells in vitro.

The natural compounds piperovatine and piperlonguminine induce autophagic cell death on Trypanosoma cruzi.

The currently available treatments for Chagas disease show limited therapeutic potential and are associated with serious side effects. Our group has been attempting to find alternative drugs isolated from natural products as a potential source of pharmacological agents against Trypanosoma cruzi. Here, we demonstrate the antitrypanosomal activity of the amides piperovatine and piperlonguminine isolated from Piper ovatum against epimastigotes and intracellular amastigotes. We also investigated the mechanisms of action of these compounds on extracellular amastigote and epimastigote forms of T. cruzi. These amides showed low toxicity to LLCMK(2) mammalian cells. By using transmission and scanning electron microscopy, we observed that the compounds caused severe alterations in T. cruzi. These alterations were mainly located in plasma membrane and mitochondria. Furthermore, the study of treated parasites labeled with Rh123, PI and MDC corroborate with our TEM data. These mitochondrial dysfunctions induced by the amides might trigger biochemical alterations that lead to cell death. Altogether, our data evidence a possible autophagic process.

Novel cytotoxic isolated from Jamaican Hyptis verticillata jacq induces apoptosis and overcomes multidrug resistance.

BACKGROUND: Adult T-cell leukemia is an aggressive hematological malignancy with a poor clinical prognosis, and a rapid resistance to chemotherapy is rapid. MATERIALS AND METHODS: Cytotoxicity assay-directed fractionation identified a novel lignan-related agent, 4-methoxy-9-(3,4,5-trimethoxyphenyl)-8, 9 - dihydrofuro[3',4':6,7]naphtho[2,3-d][1,3]dioxol-6(5H)-one (4-MTDND) from the Jamaican plant Hyptis verticillata jacq, and its effects on apoptosis, cell cycle and drug resistance were elucidated. RESULTS: The novel agent, 4-MTDND, exhibited cytotoxicity against myriad cancer types, with a wide therapeutic index of 30- to 40-fold, promoted G(2)/M arrest and up-regulated expression of pro-apoptotic proteins p53 and BAX, as well as enhanced activation of caspase-3, caspase-9 and poly (ADP ribose) polymerase, consistent with apoptosis induction. Multidrug-resistant cancer cells were as susceptible to 4-MTDND as their non-resistant control counterparts, with 4-MTDND having greater efficacy compared to standard chemotherapy agents etoposide and mitoxantrone. CONCLUSION: The novel cytotoxic agent 4-MTDND induces G(2)/M arrest and apoptosis in cancer cells possibly due to direct DNA damage or interference with topoisomerase II.

[Behaviour azole fungicide and fluconazole in Cryptococcus neoformans clinical and environmental isolates]. / Comportamento de amostras ambientais e clínicas de Cryptococcus neoformans frente a fungicidas de uso agronômico e ao fluconazol.

The activity of azole fungicides for agronomical use (epoxiconazole, difenoconazole and cyproconazole) was evaluated in comparison with the therapeutic antifungal agent fluconazole, on 23 environmental samples of Cryptococcus neoformans var neoformans isolated from pigeon feces that were collected from farms with agricultural practices using azole compounds, and on 11 clinical samples isolated from patients with cryptococcosis. Sensitivity tests were performed using the agar dilution technique. The minimum inhibitory concentration capable of inhibiting 50% of the environmental isolates (MIC 50) was 6.00 microg/ml to epoxiconazole, 1.00 microg/ml for difenoconazole, 2.00 microg/ml for cyproconazole and 64.00 microg/ml for fluconazole. Among the clinical isolates the MIC 50 values were 2.00 microg/ml, 0.38 microg/ml, 1.00 microg/ml and 16.00 microg/ml for epoxiconazole, difenoconazole, cyproconazole and fluconazole, respectively. The MIC 50 values for environmental isolates were greater than the MIC 50 values for clinical isolates. In our study, in relation to the same antifungal agent, the environmental samples presented significantly different behaviour in relation to the clinical samples (p<0.05). Differences in the MIC values (p<0.05) presented by fluconazole and the other antifungal agents for agronomical use, both in the environmental isolates and in the clinical isolates, were also observed.

Comportamento de amostras ambientais e clínicas de Cryptococcus neoformans frente a fungicidas de uso agronômico e ao fluconazol / Behaviour azole fungicide and fluconazole in Cryptococcus neoformans clinical and environmental isolates

Avaliou-se a atividade de fungicidas azólicos de uso agronômico (epoxiconazol, difenoconazol e ciproconazol) em comparação ao antifúngico de uso terapêutico fluconazol sobre 23 amostras ambientais de Cryptococcus neoformans var neoformans isoladas de fezes de pombos, as quais foram coletadas em fazendas com práticas agrícolas empregando compostos azólicos e 11 amostras clínicas isoladas de pacientes portadores de criptococose. Os testes de sensibilidade foram realizados pela técnica de diluição em agar. A concentração inibitória mínima capaz de inibir 50 por cento dos isolados ambientais (CIM 50) foi de 6,0µg/mL para epoxiconazol, 1,0µg/mL para difenoconazol, 2,0µg/mL para ciproconazol e 64,0µg/mL para fluconazol. Entre os isolados clínicos os valores de CIM 50 foram 2,0µg/mL, 0,38µg/mL, 1,0µg/mL e 16,0µg/mL para epoxiconazol, difenoconazol, ciproconazol e fluconazol, respectivamente. Os valores de CIM 50 em relação aos isolados de origem ambiental foram maiores do que os valores para os isolados de origem clínica. Em nosso estudo, frente ao mesmo antifúngico, as amostras ambientais apresentaram comportamento significativamente diferente em relação às amostras clínicas (p < 0,05). Diferenças (p<0,05) também foram observadas entre os valores de concentração inibitória apresentados pelo fluconazol e os outros antifúngicos de uso agronômico tanto no grupo dos isolados ambientais quanto clínicos.

The activity of azole fungicides for agronomical use (epoxiconazole, difenoconazole and cyproconazole) was evaluated in comparison with the therapeutic antifungal agent fluconazole, on 23 environmental samples of Cryptococcus neoformans var neoformans isolated from pigeon feces that were collected from farms with agricultural practices using azole compounds, and on 11 clinical samples isolated from patients with cryptococcosis. Sensitivity tests were performed using the agar dilution technique. The minimum inhibitory concentration capable of inhibiting 50 percent of the environmental isolates (MIC 50) was 6.00µg/ml to epoxiconazole, 1.00µg/ml for difenoconazole, 2.00µg/ml for cyproconazole and 64.00µg/ml for fluconazole. Among the clinical isolates the MIC 50 values were 2.00µg/ml, 0.38µg/ml, 1.00µg/ml and 16.00µg/ml for epoxiconazole, difenoconazole, cyproconazole and fluconazole, respectively. The MIC 50 values for environmental isolates were greater than the MIC 50 values for clinical isolates. In our study, in relation to the same antifungal agent, the environmental samples presented significantly different behaviour in relation to the clinical samples (p<0.05). Differences in the MIC values (p<0.05) presented by fluconazole and the other antifungal agents for agronomical use, both in the environmental isolates and in the clinical isolates, were also observed.

Effect of leaf essential oil from Piper solmsianum C.DC. in mice behaviour.

The essential oil from Piper solmsianum leaves and its major compound (sarisan) were tested to verify their influences upon mice behaviour. The essential oil was obtained by hydrodistillation in a modified Clevenger extractor and analysed by GC/ MS. This analysis revealed in the oil the presence of monoterpenes, sesquiterpenes and of arylpropanoids. The compound sarisan, a myristicin analogue, was isolated from the oil to perform the pharmacological tests. Emulsions of the oil and of sarisan (5.0 and 10.0% v/v) were used in the tests. Pentobarbital (30 mg/ kg s.c.) or diazepam (2.5 mg/ kg s.c.) were tested as standard drugs to verify depressant or anxiolytic effects, respectively. Both essential oil and sarisan showed to have exciting and depressant effects in the tested animals.

Effect of leaf essential oil from Piper solmsianum C.DC. in mice behaviour

The essential oil from Piper solmsianum leaves and its major compound (sarisan) were tested to verify their influences upon mice behaviour. The essential oil was obtained by hydrodistillation in a modified Clevenger extractor and analysed by GC/ MS. This analysis revealed in the oil the presence of monoterpenes, sesquiterpenes and of arylpropanoids. The compound sarisan, a myristicin analogue, was isolated from the oil to perform the pharmacological tests. Emulsions of the oil and of sarisan (5.0 and 10.0 percent v/v) were used in the tests. Pentobarbital (30 mg/ kg s.c.) or diazepam (2.5 mg/ kg s.c.) were tested as standard drugs to verify depressant or anxiolytic effects, respectively. Both essential oil and sarisan showed to have exciting and depressant effects in the tested animals

Asaricin, the main component of Ocotea opifera Mart. essential oil.

Investigation of the volatile fraction from the stem bark of Ocotea opifera Mart. led to the isolation and characterization of asaricin, a phenolic derivative with antifungal and insecticidal activity, as the main component, which is described for the first time for the genus Ocotea. The structure has been established by a study of its mono- and bidimensional NMR spectra and mass spectrometry.

[Effects of Cuban analog (SWR-104SA), a new histamine H2 receptor antagonist, on gastric acid secretion and experimental ulcer formation].

The antagonism of histamine H2-receptor by SWR-104SA (1'-bromo-N-[3-[3-(1-piperidinylmethyl) phenoxy] propyl]-spiro [1,3-dioxolane-2,9'-pentacyclo-[4.3.0.0.(2,5)0.(3,8) 0.(4,7)]nonane]-4'-carboxamide monooxalate) was estimated using the isolated guinea-pig atrium and gastric acid secretion in rats. The concentration-response curves for the positive chronotropic effect of histamine on the atrium were displaced to the right in parallel without change in the maximum response by SWR-104SA and roxatidine acetate hydrochloride (roxatidine). The pA2 values of SWA-104SA and roxatidine acetate hydrochloride were 7.27 and 7.38, respectively. The slopes of the regression line of log (DR-1) against log SWR-104SA and roxatidine concentration were 1.00 and 0.92, respectively. There was no significant difference between the two compounds with respect to the histamine H2-receptor antagonism and/or binding manner in vitro. In the rat gastric fistula model stimulated by histamine, however, antisecretory potency of SWR-104SA was 3 times less than that of roxatidine. SWR-104SA given p.o. prevented the formation of gastric lesion induced by HCl-ethanol and indomethacin dose-dependently, roxatidine also prevented its formation by HCl-ethanol, but failed to prevent that by indomethacine. These antiulcer activities of SWR-104SA were shown at the lesser doses of antisecretory activity. On the other hand, roxatidine did not prevent the ulcer formation at the same dose level of antisecretory activity. These results indicate that the antiulcer effect of SWR-104SA is not caused by the antisecretory action alone. In addition, the mucosal protective activity of SWR-104SA for HCl-ethanol induced gastric lesion was independent of endogenous prostaglandins. Moreover SWR-104SA had inhibitory effects on indomethacin-induced gastric hypermotility in rats. These actions may partly explain the gastric protection of this compound and additional mechanisms such as mucosal blood flow could be involved in the antiulcer efficacy. Consequently, it appears that SWR-104SA is a new antiulcer drug that exerts a potent cytoprotective effect in addition to its gastric antisecretory activity.

Enkephalinase inhibition facilitates sexual behavior in the male rat but does not produce conditioned place preference.

The effects of two enkephalinase inhibitors, SCH 34826 and phospho-leu-phe, on male rat sexual behavior and conditioned place preference were evaluated. SCH 34826, administered intraperitoneally, reduced the ejaculation latency to both the first and second ejaculation at a dose of 30 mg/kg. This dose also reduced the first postejaculatory interval. No other effect was obtained with this drug. Phospho-leu-phe, administered intracerebroventricularly, increased mount and intromission latency at doses of 50 and 100 micrograms. A dose of 25 micrograms reduced the latency to the first ejaculation as well as the number of preejaculatory intromissions. The postejaculatory interval was also reduced at this dose. SCH 34826, 100 and 30 mg/kg, and phospho-leu-phe, 25 micrograms, had no effect in the conditioned place preference procedure. These observations seem to suggest that there is no functionally relevant tonic release of enkephalins. Therefore, the effects obtained on sexual behavior may indicate that enkephalins are released before and during the course of sexual activity. The function of such a release could be to facilitate ejaculatory mechanisms in the way found in the present studies. Previous work has shown that ejaculation-induced reward is opioid dependent, further supporting the hypothesis of opioid release during sexual activity. Taken together, these data suggest an important role for opioids, probably enkephalins, in the physiological control of sexual behavior.