Effect of Casearia sylvestris on the obliteration of dentinal tubules and the control of dental sensitivity / Efecto de Casearia sylvestris sobre la obliteración de los túbulos dentinarios y el control de la sensibilidad dental

The present study evaluated the efficacy of the mineralizing action of Casearia sylvestris ethanolic extract on bovine dentin blocks in its pure form and in dental paste, through scanning electron microscopy. The dentin blocks were immersed in artificial saliva and incubat ed at 37°C for 7 days. Subsequently, six groups were treated with different test substances and analysed qualitatively and quantitatively at 30 and 60 days. The tests used were Kruskal - Wallis and Dunn's. Shapiro - Wilk and ANOVA. The qualitative analysis at 30 days showed a difference between the groups treated with ethanolic extract and toothpaste. Quantitatively, at 30 days, treatment with ethanolic extract of Casearia showed a greater number of open dentinal tubules. At 60 days, the difference persisted on ly for the blocks treated with toothpaste. The results obtained indicated that there is a positive relationship between the use of Casearia sylvestris and obliteration of dentinal tubules

El presente estudio evaluó la eficacia de la acción mineralizante del extracto etanólico de Casearia sylvestris sobre bloques de dentina bovina en su forma pura y en pasta dental, mediante microscopía electrónica de barrido. Los bloques de dentina se sumergieron en saliva artificial y se incubaron a 37°C durante 7 días. Posteriormente, se trataron seis grupos con diferentes sustancias de ensayo y se analizaron cualitativa y cuantitativamente a los 30 y 60 días. Las p ruebas utilizadas fueron Kruskal - Wallis y Dunn's. Shapiro - Wilk y ANOVA. El análisis cualitativo a los 30 días mostró una diferencia entre los grupos tratados con extracto etanólico y pasta dentífrica. Cuantitativamente, a los 30 días, el tratamiento con ex tracto etanólico de Casearia mostró un mayor número de túbulos dentinarios abiertos. A los 60 días, la diferencia persistió sólo para los bloques tratados con pasta dentífrica. Los resultados obtenidos indicaron que existe una relación positiva entre el us o de Casearia sylvestris y la obliteración de los túbulos dentinarios

Macro- and Microelements in the Branches of some Salix Genus Species in the Flora of Ukraine

Determination of the mineral composition is important for further study and standardization of the dried branchesof the species from the Salix (willow) family genus and extracts from the plant material. The study aimed todetermine the qualitative and quantitative content of macro- and microelements in some representatives of theSalix spp. growing in Ukraine in comparison with soils and extracts of plant material. The elemental compositionof the branches of Salix cinerea L., S. incana Schrank, S. cаprea L., S. sachalinensis F. Schmidt, S. acutifolia L.,S. fragilis L., S. caspica Pall., S. rosmarinifolia L., and S. myrsinifolia Salisb was determined using atomicemission spectrometry with photographic registration for the first time. The method used allowed us to determinethe presence of 5 macro- (К, Na, Ca, Р, Mg) and 14 microelements (Fe, Si, Мn, Al, Sr, Zn, Ni, Mo, Cu, Co, Pb,Cd, As, Hg), and the content of 14 elements were quantified in the dried branches of the Salix spp. plants. Theability to accumulate and a relatively high content of micro- and macroelements, as well as low content of toxicmicroelements (Co, Pb, Cd, As, Hg) in willow branches, allow considering them as a promising but safe sourceof biologically available microelements. There were typically not correlations between concentrations of microand macroelements in plant material and soils. The maximum levels of macroelements and microelements werefound in Salix species collected in the Kharkiv region, and in the Zakarpatye region, respectively. The elementalcomposition in the Salix sachalinensis F. Schmidt was quite different if compared with all others.

Chemical constituents from Casearia spp. (Flacourtiaceae/Salicaceae sensu lato)

ABSTRACT Chemical investigation of the leaves of Casearia gossypiosperma Briq., Salicaceae, led to the isolation of two known flavonoids, (+)-taxifolin and quercetin, the leaves of Casearia decandra Jacq. have afforded hydroquinone, the leaves of Casearia rupestris Eichler and Casearia lasiophylla Eichler have afforded a diterpene, (E)-phytol, and the leaves of C. rupestris and Casearia obliqua Spreng. have afforded sitosterol. The twigs of Casearia lasiophylla Eichler led to the isolation of two compounds (+)-pinoresinol, and N-trans-feruloyltyramine, and the twigs of C. obliqua have afforded N-trans-feruloyltyramine, N-trans-cumaroyltyramine, and cinamic acid. This is the first report of the compounds (+)-taxifolin, quercetin, hydroquinone, (+)-pinoresinol and N-trans-cumaroyltyramine from the Casearia genus.

Anatomical aspects, chemical analysis and cytotoxic effect of the essential oil from leaves of Casearia arborea (Salicaceae) / Aspectos anatómicos, análisis químico y efecto citotóxico del aceite esencial de Casearia arborea (Salicaceae)

The genus Casearia (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 200 species. In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro; including Casearia arborea (Rich.) Urb. Essential oil was obtained from the fresh leaves by hydrodistillation and analyzed by GC-MS and GC-FID. The cytotoxic effect was determined by WST-1 assay. Chemical analysis of the essential oil revealed a very diversified (n = 37 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (90.2%). These sesquiterpenes included byciclogermacrene (18.7%), germacrene D (12.1%) and α-humulene (11.5%). In addition, the essential oil demonstrated cytotoxic effects against A549 tumor cells in the concentration of 4 µg/mL (EC50) (p < 0.05).

El género Casearia (Salicáceas) se encuentra en las regiones tropicales y sub-tropicales del planeta y comprende alrededor de 200 especies. En Brasil existen 48 especies, 12 de las cuales fueron registradas en el Estado de Río de Janeiro incluyendo Casearia arborea (Rich.) Urb. El aceite esencial fue extraído de hojas frescas por hidrodestilación y analizado por GC-MS y GC-FID. El efecto citotóxico fue determinado por ensayo WST-1. Las cavidades secretorias fueron ocasionalmente encontradas tanto en la lámina foliar como en el pecíolo. El análisis químico del aceite esencial reveló una muy diversa fracción volátil (n = 37 compuestos) formada principalmente por sesquiterpenos no oxigenados (90,2%). Estos sesquiterpenos incluyen biciclogermacreno (18,7%), germacreno D (12,1%) y α-humuleno (11,5%). Además, el aceite esencial demostró efectos citotóxicos contra las células tumorales A549 en una concentración de 4µg/mL (EC50) (p < 0.05).

Casinga-cheirosa organic extract impairment over Balb-c male mice behavioral phenotype

ABSTRACT Laetia suaveolens (Poepp.) Benth., Salicaceae, popularly known as "casinga-cheirosa", "caferana", or "laranjinha", is native to Brazil but not endemic to this country. A crude organic extract was obtained from the leaves and stem and intraperitoneally administered in male Balb-c mice. Its behavioral effects were evaluated in the open field and elevated plus maze in a two-stage experiment that assessed ten different parameters related to behavior as locomotion, emotionality, and anxiety. In the first stage of the experiment, intraperitoneal the crude organic extract administration dose-dependently impaired locomotion and emotionality 30–120 min after administration. A significant decrease in defecation was observed, which was related to emotionality. No alterations in the elevated plus maze were found; thus, this apparatus was not used in the next stage of the experiment. In the second stage, the previously determined non-lethal dose of 0.1563 g/kg was intraperitoneally administered, which impaired locomotion and rearing frequency and increased immobility time. Necropsy revealed smooth intestine hemorrhage. Rutin, leucoside, nicotiflorin, guaijaverin, and astragalin were isolated from the crude organic extract. This is the first time that these compounds have been identified in L. suaveolens. In conclusion, the crude organic extract impaired locomotion and emotionality and caused hemorrhage in male Balb-c mice, indicating that its consumption can be harmful to humans and animals. The present results provide a basis for further studies on the pharmacology, toxicology, and natural product chemistry of the crude organic extract.

A new chromone glycoside from leaves of Salix matsudana / 中草药

Objective: To study the chemical constituents from the leaves of Salix matsudana. Methods: The chemical constitunents were isolated by various column chromatographic methods. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analyses. Results: Six compounds were named as 5-β-D-glucosyloxy-7-hydroxychromone (1), 5, 7-dihydroxychromone (2), (2-hydroxyphenzyl)-O-β-D-glucopyranoside (3), vanillic acid-4-O-β-D-glucoside (4), cyclo (val-pro) (5), and gallic acid (6). Conclusion: Compound 1 is a new compound named matsudoside A, compounds 2-6 are obtained from this plant for the first time.

Avaliação da atividade antibacteriana das cascas das raízes de Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato) / Validación de la actividad antibacteriana de corteza de raíces de Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato) / Validation of the antibacterial activity of root bark of Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato)

Introdução: Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato), espécie conhecida popularmente como sucará, açucará ou espinho-de-judeu é uma árvore de pequeno porte, que se desenvolve em regiões de vegetação de mata ciliar. Objetivos: avaliar o potencial antibacteriano de frações e substância isolada das cascas das raízes de Xylosma ciliatifolia sobre Staphylococcus aureus, Staphylococcus epidermidis, Salmonella typhimurium e Escherichia coli. Métodos: o material vegetal seco foi extraído em etanol 96 % em aparelho de Soxhlet com aquecimento. O extrato etanólico obtido foi submetido à partição líquido-líquido em hexano, clorofórmio e acetato de etila e, após, concentrados em banho-maria, obtendo-se, respectivamente, as frações hexano, clorofórmio e acetato de etila. Por cromatografia em coluna da fração hexano obteve-se o composto ugandensidial. A atividade antibacteriana foi avaliada pelo ensaio da difusão em disco e pela medida da concentração inibitória mínima. Resultados: a atividade antibacteriana foi evidenciada nas frações hexano e clorofórmio sobre as bactérias: Staphylococcus aureus e Staphylococcus epidermidis. O composto ugandensidial mostrou-se ainda mais bioativo sobre as cepas Staphylococcus aureus e Staphylococcus epidermidis com inibição do crescimento bacteriano na concentração de 62,5 µg/mL. Conclusão: os ensaios antibacterianos realizados neste trabalho com as cascas das raízes de Xylosma ciliatifolia foram capazes de inibir o crescimento de duas cepas gram positivas. Estes resultados contribuem para o conhecimento fitoquímico dessa espécie vegetal que ainda não apresenta dados em literaturas científicas.

Introducción: Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato), especie conocida en la población como sucará, acucará o espinho-de-judeu es un árbol de pequeño porte, que se desarrolla en las regiones de vegetación de ribera. Objetivos: evaluar la actividad antibacteriana de las fracciones y sustancias aisladas de la corteza de raíces de Xylosma ciliatifolia sobre Staphylococcus aureus, Staphylococcus epidermidis, Salmonella typhimurium y Escherichia coli. Métodos: el material seco se extrajo en etanol 96 % en un aparato de Soxhlet con calentamiento. El extracto etanólico obtenido se sometió a partición líquido-líquido en hexano, cloroformo y acetato de etilo; después se concentró en un baño de agua, produciendo, respectivamente, las fracciones hexano, cloroformo y acetato de etilo. Por cromatografía de columna de la fracción hexano se obtuvo el compuesto ugandensidial. La actividad antibacteriana se evaluó mediante la prueba de la difusión en disco y mediante la medición de la concentración inhibitoria mínima. Resultados: la actividad antibacteriana se observó en las fracciones hexano y cloroformo sobre las bacterias: Staphylococcus aureus y Staphylococcus epidermidis. El compuesto ugandensidial resultó aún más bioactivo sobre las bacterias Staphylococcus aureus y Staphylococcus epidermidis, con inhibición del crecimiento bacteriano en la concentración de 62,5 µg/mL. Conclusiones: los ensayos antibacterianos realizados en este estudio con corteza de raíces de Xylosma ciliatifolia fueron capaces de inhibir el crecimiento de ambas cepas grampositivas. Estos resultados contribuyen al conocimiento fitoquímico de esa especie vegetal que aún no presenta datos en la literatura científica.

Introduction: Xylosma ciliatifolia (Clos) Eichler (Flacourtiaceae/Salicaceae sensu lato), a species known by the general public as sucara, acucara or espinho-de-judeu, is a small tree growing in riverside areas. Objectives: evaluate the antibacterial activity of fractions and substances isolated from root bark of Xylosma ciliatifolia against Staphylococcus aureus, Staphylococcus epidermis, Salmonella typhimurium and Escherichia coli. Methods: the dry material was extracted into 96% ethanol in a Soxhlet device with heating. The ethanolic extract thus obtained was subjected to liquid-liquid partition in hexane, chloroform and ethyl acetate. It was then concentrated in a water bath to obtain hexane, chloroform and ethyl acetate fractions, respectively. The ugandensidial compound was obtained by column chromatography of the hexane fraction. Antibacterial activity was evaluated by the disc diffusion test and by measurement of the minimum inhibitory concentration. Results: antibacterial activity against Staphylococcus aureus and Staphylococcus epidermis was observed in the hexane and chloroform fractions. The ugandensidial compound was found to be even more bioactive against the bacteria Staphylococcus aureus and Staphylococcus epidermis, with inhibition of bacterial growth in the 62.5 µg/mL concentration. Conclusions: the antibacterial assays conducted in this study with root bark of Xylosma ciliatifolia revealed inhibition in the growth of both gram-positive strains. These results provide phytochemical information about this plant species, for which there are no data in the scientific literature.

Chemical composition and cytotoxic activity of the essential oil from the leaves of Casearia lasiophylla

The essential oil obtained by hydrodistillation from fresh leaves of Casearia lasiophylla Eichler, Salicaceae, was analyzed by gas capillary (GC/FID and GC/MS). The cytotoxicity of the leaves essential oil was tested in vitro againstU251 (glioma), UACC-62 (melanoma), MCF-7 (breast), NC1-ADR/RES (ovarian-resistant), NCI-H460 (lung), PC03 (prostate), OVCAR-3 (ovarian), HT-29 (colon) and K562 (leukemia) human cancer cells and against VERO (no cancer cell). The yield of oil was 0.02 percent. Fifty two compounds were identified, representing 87.1 percent of the total of the oil. The main components were identified as germacrene D (18.6 percent), β-caryophyllene (14.7 percent), δ-cadinene (6.2 percent), and α-cadinol (5.4 percent). The oil exhibited antiproliferative activity against all cell lines (TGI<100 µg/mL), with exception of NCI-H460 cell line (TGI 191.31 µg/mL). The highest activity was observed against UACC-62 (TGI 7.30 µg/mL), and K562 (TGI 7.56 µg/mL) cell lines. The observed activity could be related to high content of germacrene D and β-caryophyllene, compounds known as cytotoxic.

Identificação de diterpenos clerodânicos em diferentes órgãos de Casearia sylvestris Swartz / Identification of clerodane diterpenes in different organs of Casearia sylvestris Swartz

Casearia sylvestris Swartz (Salicaceae) é uma espécie vegetal arbórea ou arbustiva, disseminada pelo território brasileiro e bastante utilizada em nossa medicina popular. Diversos diterpenos clerodânicos bioativos típicos do gênero Casearia foram isolados desta espécie (ex. casearinas e casearvestrinas). O objetivo principal deste trabalho foi avaliar a presença de diterpenos clerodânicos em diferentes órgãos de C. sylvestris, utilizando técnicas analíticas cromatográficas e espectroscópicas. Os extratos dos diferentes órgãos foram analisados por cromatografia em camada delgada, cromatografia líquida de alta eficiência com detector de arranjo de diodos e ressonância magnética nuclear de 1H. Nas análises cromatográficas foram utilizados diterpenos clerodânicos isolados de C. sylvestris como padrões, incluindo rel-19S-acetóxi- 18R- butanoilóxi- 18,19- epóxi -6S -hidróxi -2R- (2-metilbutanoilóxi) -5S, 8R, 9R, 10S -cleroda-3,13(16),14-trieno, isolado do caule pela primeira vez. Foram obtidos perfis fitoquímicos dos órgãos avaliados. Os resultados indicaram a presença de diterpenos clerodânicos em toda planta, prevalecendo em maior número nas folhas. Além disso, os resultados também indicam que em caules, flores e raízes há predomínio de diterpenos clerodânicos com padrão diênico diferente daqueles obtidos de folhas de C. sylvestris.

Casearia sylvestris Swartz (Salicaceae) is a tree or shrub distributed widely in Brazil, where it is used in popular medicine. Several bioactive clerodane diterpenes typical of Casearia have been isolated from this species (e.g. casearins and casearvestrins). The main objective of this study was to identify clerodane diterpenes in various organs of C. sylvestris, using chromatographic and spectroscopic analytical techniques. The extracts of the different plant parts were analyzed by thin layer chromatography, high performance liquid chromatography with diode array detector and 1H nuclear magnetic resonance. In the chromatographic analysis, clerodane diterpenes isolated from C. sylvestris were used as standards, including rel-19Sacetóxi- 18R- butanoilóxi-18,19- epóxi -6S -hidróxi -2R- (2-metilbutanoilóxi) -5S, 8R, 9R, 10S -cleroda-3,13(16),14- triene, isolated for the first time from the stems. Phytochemical profiles of the organs were produced, which indicated the presence of clerodane diterpenes in all parts of the plant, notably in the leaves. The results also suggest that the main clerodane diterpenes in the stems, flowers and roots had conjugated double-bond patterns that differed from those found in the leaves.

Alelopatia de Joanesia princeps Vell. e Casearia sylvestris Sw. sobre espécies cultivadas / Allelopathy of Joanesia princeps Vell. and Casearia sylvestris Sw. on the cultivated species

Metabólitos secundários produzidos em algumas plantas podem provocar alterações no desenvolvimento de outras plantas ou até mesmo de outros organismos. O objetivo do trabalho foi avaliar o efeito alelopático de extratos aquosos de folhas de Joanesia princeps e Casearia sylvestris na germinação e no desenvolvimento inicial de plântulas de Brassica oleracea cv. capitata, Lactuca sativa cv. grand rapids e Lycopersicum esculentum. Foram testadas seis concentrações de cada extrato (10, 30, 50, 70, 90 e 100 por cento) além do controle água destilada. Os extratos aquosos de J. princeps e C. sylvestris reduziram e, ou inibiram significativamente o percentual de geminação das sementes, o crescimento inicial da parte aérea e do sistema radicular de todas as espécies cultivadas e causaram severas anormalidades nas plântulas. Os resultados indicam a existência de potencial alelopático de J. princeps e C. sylvestris.

Secondary metabolites produced in some plant species can provoke development changes in other plants or even in other organisms. The objective of this work was to identify the possible allelopathic effects of the aqueous extracts of Joanesia princeps and Casearia sylvestris leaves in the germination and initial growth of Brassica oleracea cv. capitata, Lactuca sativa cv. grand rapids and Lycopersicum esculentum seedlings. Six concentrations of each extract (10, 30, 50, 70, 90 and 100 percent) were tested besides distilled water control. The aqueous extracts of J. princeps and C. sylvestris significantly reduced the seed germination percentage, initial growth of the aerial section and root system of all cultivated species in which they caused severe seedlings damage. The results indicate existence of allelopathic potential in J. princeps and C. sylvestris.

Protease inhibition activity of extracts from Salicaceae species from Brazilian Cerrado and Atlantic Rain Forest and of an enriched fraction of clerodane diterpenes (casearins) / Atividade biológica de uma fração enriquecida de diterpenos clerodânicos (casearinas) e de extratos de espécies de Salicaceae do Cerrado e Mata Atlântica em um ensaio de inibição de proteases

Considering the traditional use of Casearia sylvestris Sw., Salicaceae, to threat gastric injuries and the pre-clinical studies showing its efficacy we aimed to screen other species to explore the biological activity of some species of this family. For this, we used a protease inhibition assay as a model for searching gastric anti-ulcer plant extracts. The ethanolic and aqueous extracts from branches and leafs of C. gossypiosperma, C. decandra and C. rupestris showed high percentage inhibition of pepsin, approximately 50 percent, with 1 μg/mL concentration. Curiously, C. obliquoa and Flacourtia ramontchi did not inhibit pepsin, but its most apolar extract showed inhibitory activity in the subtilisin assay. The enriched fraction of clerodane diterpenes inhibited the activity (42.75 percent) of pepsin with 1 ug/mL, but it did not inhibit subtilisin (23.76 percent). The results obtained with apolar and polar extracts from branches and leaves of some species of Salicaceae showed a different pattern of inhibition of two proteases, the aspartic pepsin and the serinic subtilisin, related with different biological activities. The results with the enriched fraction of clerodane diterpenes suggests that the activity observed with the C. sylvestris may be related with the presence of these substances in the crude extract.

Considerando o uso popular de Casearia sylvestris Sw., Salicaceae, para o tratamento de problemas gástricos e resultados pré-clínicos que mostraram potencial atividade anti-ulcerogênica, foi realizado um screening farmacológico para avaliar a atividade biológica de outras espécies de Salicaceae. Para isso, foi utilizado um ensaio de inibição de proteases como um modelo farmacológico molecular para screening de extratos com atividade anti-ulcerogênica. Os extratos etanólico e aquoso dos galhos e folhas de C. gossypiosperma, C. decandra e C. rupestris mostraram inibição da atividade da pepsina em aproximadamente 50 por cento com a concentração de 1 μg/mL. Curiosamente, C. obliquoa e Flacourtia ramontchi não apresentaram atividade sobre a pepsina, mas seus extratos mais apolares mostraram atividade inibitória sobre a subtilisina. A fração enriquecida de diterpenos clerodânicos mostrou atividade inibitória (42,75 por cento) sobre a pepsina com a concentração de 1 μg/mL, mas não sobre a subtilisina (23,76 por cento). Os resultados obtidos com os extratos e folhas das espécies testadas mostraram um padrão de atividade diferente sobre os dois tipos de proteases, a pepsina e a subtilisina, as quais estão relacionadas com diferentes tipos de atividades biológicas. Ainda mais, os resultados com a fração enriquecida de diterpenos clerodânicos sugerem que estas substâncias podem estar relacionadas com a atividade do extrato bruto de C. sylvestris.

Estudo de toxicologia clínica de um fitoterápico contendo Passiflora incarnata L., Crataegus oxyacantha L., Salix alba L. em voluntários saudáveis / Clinical toxicology study of an herbal medicine with Passiflora incarnata L., Crataegus oxyacantha L., Salix alba L. in healthy volunteers

O Pasalix® é um produto fitoterápico contendo uma associação de três plantas medicinais: Passiflora incarnata, Salix alba e Crataegus oxyacantha. Sua principal indicação é para o tratamento da ansiedade e insônia. O objetivo desse estudo foi avaliar a toxicologia clínica dessa formulação em voluntários saudáveis. Para isso realizou-se um ensaio clínico não aleatório, aberto, com 24 voluntários sadios do sexo masculino, que receberam ambulatorialmente dois (2) comprimidos revestidos do fitoterápico duas vezes ao dia, durante 28 dias ininterruptos. Os voluntários foram incluídos no estudo somente quando considerados saudáveis após avaliação clínica, exame físico e exames laboratoriais que antecederam o estudo. A avaliação laboratorial incluiu análise hematológica, bioquímica e sorológica. A avaliação clínica e laboratorial foi repetida após a 1ª, 2ª, 3ª e 4ª semanas de tratamento e 7 dias após a última administração. O Pasalix® foi bem tolerado pelos 24 voluntários não apresentando eventos adversos graves. Os exames clínicos, eletrocardiográficos e laboratoriais efetuados antes, durante e após o ensaio não evidenciaram sinais de toxicidade nos diversos órgãos e sistemas avaliados, confirmando a segurança da preparação para utilização em ensaios de eficácia terapêutica.

Pasalix® is an herbal medicine containing a combination of three medicinal plants: Passiflora incarnata, Salix alba and Crataegus oxyacantha. Its main indication is to treat anxiety and insomnia. The purpose of this study was to evaluate the clinical toxicology of that formulation in healthy volunteers. For this, a non-randomized open clinical trial was conducted with 24 healthy male volunteers, who received two (2) coated tablets of the herbal medicine twice a day for 28 uninterrupted days. The volunteers were included in the study only when considered healthy after clinical assessment, physical examination and laboratory tests which preceded the study. The laboratory tests included: hematological, biochemical and serological analysis. The clinical and laboratory evaluation was repeated after the 1st, 2nd, 3rd and 4th weeks of the treatment and 7 days after the last administration. Pasalix® was well tolerated by the 24 volunteers, and it has showed no serious adverse events. The clinical, laboratory, and electrocardiographic data assessed before, during and after the test showed no signals of toxicity in various organs and systems evaluated, confirming the safety of the preparation for use in trials of therapeutic efficacy.