Estudo fitoquímico, morfoanatômico, atividades biológicas e propriedades antioxidantes da espécie Myrcia hatschbachii D. Legrand, Myrtaceae

Orientadora : Profª Drª Marilis Dallarmi MiguelCoorientador : Prof. Dr. Obdulio Gomes MiguelDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa : Curitiba, 21/02/2018Inclui referências : p. 128-138Resumo : A especie Myrcia hatschbachii D. Legrand (Myrtaceae) e nativa e endemica do Brasil, tendo ocorrência confirmada nos estados do Sul. O objetivo deste trabalho foi realizar o estudo fitoquímico, morfoanatômico e avaliar as propriedades antioxidantes e atividades biológicas, visto ser apenas uma das muitas especies da biodiversidade brasileira sem respaldo cientifico documentado na literatura. O material vegetal foi coletado em Curitiba, Paraná, Brasil e identificado por comparação no Museu Botânico Municipal. O estudo morfoanatômico foi realizado por microscopia optica e microscopia eletrônica de varredura, além da realização de testes histoquímicos. A folha apresentou estômatos paracíticos e nervura central com formato côncavo-convexo. Houve a presença de tricomas tectores, drusas de oxalato de cálcio e compostos fenólicos nas folhas, pecíolos e galhos. No screening fitoquímico foi observada a presença de compostos fenólicos e esteroides e/ou triterpenos. O óleo essencial foi extraído por Clevenger, resultou em um rendimento de 0,17% e seus compostos majoritários foram trans-Calameneno, E-Cariofileno e Espatulenol. A extração das folhas e galhos foi realizada por Soxhlet e apos o fracionamento, resultaram-se as frações hexano, clorofórmio, acetato de etila e remanescente. A partir da fração folha acetato de etila foi isolado e identificado o acido gálico. Os testes antioxidantes por formação do complexo fosfomolibdenio e redução do radical DPPHo apresentaram significativa atividade, com destaque para as frações folha e galho acetato de etila. No ensaio de toxicidade in vitro frente a Artemia salina, o óleo essencial foi considerado moderadamente toxico. Na alelopatia, houve inibição do crescimento do hipocótilo e radícula nos extratos brutos e frações de folhas e galhos, além do óleo essencial. Obteve-se atividade hemolitica na faixa de 50% na concentração de 1000 yg/mL da fração folha hexano e óleo essencial. Considerando a presença do constituinte acido gálico, as propriedades antioxidante e alelopática demonstradas pelos extratos brutos e frações, bem como a toxicidade do óleo essencial, estudos de aplicações diversas podem ser aprofundados posteriormente para o enriquecimento da pesquisa desta espécie. Palavras-chaves: Óleo essencial. Acido gálico. Toxicidade. Alelopatia. Hemólise.Abstract : The species Myrcia hatschbachii D. Legrand (Myrtaceae) is native and endemic to Brazil, having confirmed occurrence in the Southern States. The aim of this research was to realize phytochemical, morphological and anatomical studies, and evaluate antioxidant properties and biological activities, as this is only one of many species that compose brazilian biodiversity without any scientific study documented in literature. The plant material was collected in Curitiba, Parana, Brazil and identified by comparison in Museu Botanico Municipal (Municipal Botanical Museum). The morphological and anatomical studies were performed by using optical and electronic microscopy, on top of histochemical tests. The leaf presented paracitic stomata and central nervure with concave-convex format. Tector trichomes, calcium oxalate druses and phenolic compounds were present on leaves, stems and branches. By using phytochemical screening, it was observed that phenolic compounds and steroids and/or triterpenes were present. The essential oil was extracted by using Clevenger apparatus, resulting in yield of 0.17% and its main compounds were trans- Calamenene, E-Caryophylene and Spathulenol. Extractions from leaves and branches were obtained via Soxhlet and after fractioning, resulted in hexane, chloroform, ethyl acetate and remaining hydroalcoholic fractions. From the leaf ethyl acetate fraction was isolated and identified gallic acid. The antioxidant capacity by formation of phosphomolybdenum complex and DPPHo radical scavenging method presented significant activity, with special emphasis in leaf and branch ethyl acetate fraction. In the test for in vitro toxicity towards Artemia salina, the essential oil was considered moderately toxic. In the allelopathic study, there was growth inhibition in hypocotyl and radicle in crude extracts and fractions from leaves and branches, and also essential oil. Hemolytic activity was found in the 50% range at 1000 yg/mL of leaf hexane fraction and essential oil. Considering the presence of the gallic acid, the antioxidant and allelopathic properties demonstrated by the crude extracts and fractions, as well as the essential oil toxicity, several applications studies can be further developed to enrich the research of this species. Keywords: Essential Oil. Gallic acid. Toxicity. Allelopathy. Hemolisis

Contribution to the identification of the species Myrcia hatschbachii D. Legrand (Myrtaceae): anatomical and histochemical analyses

The genus Myrcia is used in folk medicine to treat diabetes. The plants used in folk medicine require morphological and anatomical references to attest to its authenticity. This is the first report of the microscopic study of Myrcia hatschbachii. In this sense, the aim of the present study was to describe the anatomical characteristics, in order to contribute to the species' identification. For anatomical and surface analyses free-hand sections were prepared and observed using optical microscopy, simultaneously some materials were processed and observed under scanning electron microscopy. In addition, histochemical tests were performed. The anatomical features described here correspond with previously reported features found in other species of the genus Myrcia. Furthermore, the following anatomical markers were observed in Myrcia hatschbachii: druses in the spongy parenchyma, concave-convex midrib, heart-shaped petiole, oval-shaped stem, C-shaped vascular bundle in the midrib and open arc shape with invaginated ends in the petiole; phenolic compounds in the phloem and lignified cells in the pith. The results of anatomical and histochemical analyses provide micromorphological and microchemical features that can help in the taxonomy and proper identification of the species

Chemical composition, phytotoxic potential, biological activities and antioxidant properties of Myrcia hatschbachii D. Legrand essential oil

Myrcia hatschbachii D. Legrand (Myrtaceae) is a native and endemic species from Brazil. This study investigated the essential oil composition, phytotoxic potential, in vitro toxicity, antioxidant properties and antibacterial activity of species. Chromatographic analysis of essential oil identified trans-calamanene, (E)-caryophyllene and spathulenol as major components. Antioxidant capacity was determined by the DPPH• scavenging method and phosphomolybdenum complex formation assay. Antibacterial activity was evaluated using the Minimal Inhibitory Concentration test, demonstrating that the essential oil was active toward Enterococcus faecalis and Staphylococcus aureus. Phytotoxic activity of essential oil was analyzed by testing interference on germination and growth of Lactuca sativa, demonstrating significant inhibition of the hypocotyls and radicles of seeds. Preliminary toxicity studies were determined using Artemia salina, resulting in an LC50 of 409.92 µg/mL, and through hemolytic activity. The results of the phytotoxic activity point to a possible application for Myrcia hatschbachii in the development of natural herbicides and the in vitro toxicity assays suggests the performance of antitumoral activity tests, having in mind the prospection of antineoplastic drugs

Composition, antioxidant properties, and biological activities of the essential oil extracted from Ocotea diospyrifolia (Meisn.) Mez.

The article has aimed to characterize the essential oil extracted from Ocotea diospyrifolia (Meisn.) Mez. leaves, in terms of its chemical composition and antioxidant, hemolytic, and phytotoxic potentials, as well as its toxicity against Artemia salina. The major constituents identified by CG-MS were δ-elemene, spathulenol, and β-atlantol. When screened for potential biological activities, the essential oil presented low toxicity against Artemia salina, and a capacity of lysing red blood cells. Also, the evaluation of its in vitro antioxidant activity, using the phosphomolybdenum method, showed better results when compared to butylhydroxytoluene (BHT) and rutin. In conclusion, the results obtained showed certain in vitro toxicity, leading to an interesting target for cytotoxicity evaluations of carcinoma cells

Infecção por múltiplos tipos de Papilomavirus humano em mulheres jovens sexualmente ativas

Modelo do estudo: transversal. Objetivo do estudo: estimar a frequência e a distribuição dos tipos de Papilomavírus humano (HPV) em mulheres jovens sexualmente ativas. Metodologia: foram coletadas amostras do canal vaginal e endocérvice de 158 mulheres, com idade entre 18 e 35 anos, por meio da técnica de autocoleta. O DNA-HPV foi extraído e amplificado pela técnica de PCR end point utilizando os primers consensus PGMY09/11. A genotipagem foi executada pela técnica de PCR tipo específico (TSPCR) e por análise dos fragmentos obtidos com o uso de enzimas de restrição (RFLP). Resultados: DNA-HPV foi detectado em 23% das amostras, 92% foram genótipos de alto risco oncogênico (HR), sendo prevalente o HPV45, seguido do HPV16 e 31. Infecções por múltiplos tipos de HPV foram detectadas em 35% das amostras, demonstrando infecção por mais de dois tipos em 22,2% das genotipadas. Maior frequência do DNA-HPV foi observada entre mulheres com idade ≤ 25 anos. Conclusão: Este estudo demostrou alta frequência da infecção pelo HPV em mulheres jovens sexualmente ativas, assim como uma alta prevalência de infecção múltipla com tipos de alto risco oncogênicoStudy design: cross. Study objective: estimate the frequency and distribution types of human papillomavirus (HPV) in sexually active young women. Methods: samples of vaginal and endocervical canal of 158 women, aged between 18 and 35 years were collected through self-collection technique. The HPV DNA was extracted and amplified by PCR end point using the consensus primers PGMY09/11 and genotyping was performed by PCR specific type (TS-PCR) and by analysis of the fragments obtained from the use of restriction enzymes (RFLP). Results: HPV DNA was detected in 23% of participants, 92% were high-risk HPV genotypes (HR), being the prevalent HPV45, followed by HPV16 and 31 infections with multiple HPV types were detected in 35% of samples, indicating infection by more than two types in 22.2% of the genotyped. Higher frequency of HPV DNA was detected in women ≤ 25 old. Conclusion: this demonstrated a high frequency of HPV infection in sexually active young women, as well as a high prevalence of infection with multiple types of high oncogenic ris

Estudo fitoquímico, morfoanatômico, atividades biológicas e propriedades antioxidantes da espécie Myrcia hatschbachii D. Legrand, Myrtaceae

Orientadora : Profª Drª Marilis Dallarmi MiguelCoorientador : Prof. Dr. Obdulio Gomes MiguelDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa : Curitiba, 21/02/2018Inclui referências : p. 128-138Resumo : A especie Myrcia hatschbachii D. Legrand (Myrtaceae) e nativa e endemica do Brasil, tendo ocorrência confirmada nos estados do Sul. O objetivo deste trabalho foi realizar o estudo fitoquímico, morfoanatômico e avaliar as propriedades antioxidantes e atividades biológicas, visto ser apenas uma das muitas especies da biodiversidade brasileira sem respaldo cientifico documentado na literatura. O material vegetal foi coletado em Curitiba, Paraná, Brasil e identificado por comparação no Museu Botânico Municipal. O estudo morfoanatômico foi realizado por microscopia optica e microscopia eletrônica de varredura, além da realização de testes histoquímicos. A folha apresentou estômatos paracíticos e nervura central com formato côncavo-convexo. Houve a presença de tricomas tectores, drusas de oxalato de cálcio e compostos fenólicos nas folhas, pecíolos e galhos. No screening fitoquímico foi observada a presença de compostos fenólicos e esteroides e/ou triterpenos. O óleo essencial foi extraído por Clevenger, resultou em um rendimento de 0,17% e seus compostos majoritários foram trans-Calameneno, E-Cariofileno e Espatulenol. A extração das folhas e galhos foi realizada por Soxhlet e apos o fracionamento, resultaram-se as frações hexano, clorofórmio, acetato de etila e remanescente. A partir da fração folha acetato de etila foi isolado e identificado o acido gálico. Os testes antioxidantes por formação do complexo fosfomolibdenio e redução do radical DPPHo apresentaram significativa atividade, com destaque para as frações folha e galho acetato de etila. No ensaio de toxicidade in vitro frente a Artemia salina, o óleo essencial foi considerado moderadamente toxico. Na alelopatia, houve inibição do crescimento do hipocótilo e radícula nos extratos brutos e frações de folhas e galhos, além do óleo essencial. Obteve-se atividade hemolitica na faixa de 50% na concentração de 1000 yg/mL da fração folha hexano e óleo essencial. Considerando a presença do constituinte acido gálico, as propriedades antioxidante e alelopática demonstradas pelos extratos brutos e frações, bem como a toxicidade do óleo essencial, estudos de aplicações diversas podem ser aprofundados posteriormente para o enriquecimento da pesquisa desta espécie. Palavras-chaves: Óleo essencial. Acido gálico. Toxicidade. Alelopatia. Hemólise.Abstract : The species Myrcia hatschbachii D. Legrand (Myrtaceae) is native and endemic to Brazil, having confirmed occurrence in the Southern States. The aim of this research was to realize phytochemical, morphological and anatomical studies, and evaluate antioxidant properties and biological activities, as this is only one of many species that compose brazilian biodiversity without any scientific study documented in literature. The plant material was collected in Curitiba, Parana, Brazil and identified by comparison in Museu Botanico Municipal (Municipal Botanical Museum). The morphological and anatomical studies were performed by using optical and electronic microscopy, on top of histochemical tests. The leaf presented paracitic stomata and central nervure with concave-convex format. Tector trichomes, calcium oxalate druses and phenolic compounds were present on leaves, stems and branches. By using phytochemical screening, it was observed that phenolic compounds and steroids and/or triterpenes were present. The essential oil was extracted by using Clevenger apparatus, resulting in yield of 0.17% and its main compounds were trans- Calamenene, E-Caryophylene and Spathulenol. Extractions from leaves and branches were obtained via Soxhlet and after fractioning, resulted in hexane, chloroform, ethyl acetate and remaining hydroalcoholic fractions. From the leaf ethyl acetate fraction was isolated and identified gallic acid. The antioxidant capacity by formation of phosphomolybdenum complex and DPPHo radical scavenging method presented significant activity, with special emphasis in leaf and branch ethyl acetate fraction. In the test for in vitro toxicity towards Artemia salina, the essential oil was considered moderately toxic. In the allelopathic study, there was growth inhibition in hypocotyl and radicle in crude extracts and fractions from leaves and branches, and also essential oil. Hemolytic activity was found in the 50% range at 1000 yg/mL of leaf hexane fraction and essential oil. Considering the presence of the gallic acid, the antioxidant and allelopathic properties demonstrated by the crude extracts and fractions, as well as the essential oil toxicity, several applications studies can be further developed to enrich the research of this species. Keywords: Essential Oil. Gallic acid. Toxicity. Allelopathy. Hemolisis

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer