Estudo morfo-anatômico e fitoquímico de Ocotea puberula (Rich.) Nees, família Lauraceae, da região dos Campos Gerais, Ponta Grossa, Paraná

Orientador : Tomoe NakashimaCoorientadores : Yedo Alquini e Jocélia Lago JansenDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências BiológicasResumo: Ocotea puberula (Rich.) Nees é uma espécie arbórea nativa, que ocorre por toda América tropical e subtropical, em quase todas as formações florestais. É conhecida como guaicá, canela-guaicá e canela-sebo, sendo utilizada por comunidades indígenas no tratamento de tumores e afecções da pele. O objetivo deste trabalho é avaliar a anatomia da folha, caule e casca de O. puberula e estudar a constituição química da espécie. A coleta do material botânico foi realizada no Parque Municipal da Boca da Ronda, no Município de Ponta Grossa, Estado do Paraná. Para os ensaios botânicos, foram preparadas lâminas permanentes e semipermanentes, utilizando-se azul de toluidina e fucsina básica mais azul de astra para a coloração. Nos ensaios fitoquímicos, realizou-se a marcha sistemática fitoquímica, a extração do óleo essencial com o aparelho de Clevenger e a cromatografia capilar de alta resolução acoplada ao detector de massas do óleo essencial das cascas de O. puberula. Folha, caule e casca de O. puberula apresentaram características anatômicas inerentes à família Lauraceae. A anatomia foliar revelou a ocorrência de estômatos paracíticos e tricomas tectores unicelulares na face abaxial, mesofilo dorsiventral, nervura mediana biconvexa, com sistema vascular constituído de feixe único do tipo colateral em arco aberto. O caule apresentou-se em estrutura secundária incipiente. Na casca verificou-se a periderme, o córtex formado por células parenquimáticas de paredes delgadas, o periciclo descontínuo, o sistema floemático percorrido por raios parenquimáticos com 2-3 fileiras de células e as fibras liberianas agrupadas em pequenos conjuntos. Na estrutura anatômica do limbo foliar, da nervura mediana, do pecíolo, do caule e da casca foram observados idioblastos formados por células secretoras de substâncias lipofílicas e mucilaginosas. Os ensaios fitoquímicos demonstraram a presença de alcalóides, taninos, aminogrupos, esteróides e/ou triterpenos, ácidos fixos e glicosídeos saponínicos. O rendimento percentual de óleo essencial nas cascas foi de 0,15% (v/p). A análise por GC/MS-MSD revelou a presença do espatulenol (15,73%), 2-beta-pineno (9,70%), biciclogermacreno (9,38%), germacreno D (6,62%), alfa-pineno (6,26%), trans-muurolol (5,72%), limoneno (5,50%), nerolidol (4,42%), calareno (3,64%) e delta-cadineno (3,08%). Os resultados obtidos são pontos de partida para a realização de estudos farmacológicos e microbiológicos com a espécie em questão.Abstract: Ocotea puberula (Rich.) Nees is a native arboreal species. It occurs all over tropical and subtropical America, in almost all the forest formations. It is known as guaicá, canela-guaicá and canela-sebo, being used by indigenous communities to treat skin diseases and tumors. This work aims at evaluating the anatomy of the O. puberula's leaf, stem and bark and studying chemical composition of the species. The botanical material was collected from the Municipal Park of Boca da Ronda, in Ponta Grossa, State of Paraná. Permanent and semi permanent slides were prepared for the botanical assays. The stain was made using toluidine blue and fuchsin basic plus astra blue. Phytochemical studies involved the chemical screening, the essential oil extraction and the gas-liquid chromatographic/mass detector analysis of the bark essential oil of O. puberula. Leaf, stem and bark of O. puberula presented inherent anatomical characteristics to the Lauraceae. The leaf anatomy showed paracytic stomata and unicellular trichomes on the abaxial surface, dorsiventral mesophyll, biconvex midrib constituted by a collateral bundle in open arch. The stem presented incipient secondary structure. The bark demonstrated periderm, cortex formed by thin-walled parenchyma cells, discontinuous pericycle, phloem system crossed by a 2-3 cell-parenchyma rays and small groups of phloem fibers. Oil and mucilage cells were found in leaf, stem and bark anatomical structures. Phytochemical assays demonstrated the presence of alkaloids, tannins, aminogroups, steroids and/or triterpenoids, fixed acids and saponin glycosides. Barks yielded 0.15% (v/w) of essential oil. The GC/MS-MSD analysis showed spathulenol (15.73%), 2-beta-pinene (9.70%) bicyclogermacrene (9.38%), germacrene D (6.62%), alpha-pinene (6.26%), traws-muurolol (5.72%), limonene (5.50%), nerolidol (4.42%), calarene (3.64%) and delta-cadinene (3.08%). The obtained results are starting points for pharmacological and microbiological studies

A simple HPLC method for the determination of halcinonide in lipid nanoparticles: development, validation, encapsulation efficiency, and in vitro drug permeation

Halcinonide is a high-potency topical glucocorticoid used for skin inflammation treatments that presents toxic systemic effects. A simple and quick analytical method to quantify the amount of halcinonide encapsulated into lipid nanoparticles, such as polymeric lipid-core nanoparticles and solid lipid nanoparticles, was developed and validated regarding the drug's encapsulation efficiency and in vitro permeation. The development and validation of the analytical method were carried out using the high performance liquid chromatography with the UV detection at 239 nm. The validation parameters were specificity, linearity, precision and accuracy, limits of detection and quantitation, and robustness. The method presented an isocratic flow rate of 1.0 mL.min-1, a mobile phase methanol:water (85:15 v/v), and a retention time of 4.21 min. The method was validated according to international and national regulations. The halcinonide encapsulation efficiency in nanoparticles was greater than 99% and the in vitro drug permeation study showed that less than 9% of the drug permeated through the membrane, indicating a nanoparticle reservoir effect, which can reduce the halcinonide's toxic systemic effects. These studies demonstrated the applicability of the developed and validated analytical method to quantify halcinonide in lipid nanoparticles

DEVELOPMENT AND VALIDATION OF AN HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE DETERMINATION OF 17-Β ESTRADIOL IN POLYMERIC NANOPARTICLES

Objective: A simple high-performace liquid chromatography method was developed and validated to determine 17-β estradiol in poly (ε-caprolactone) nanocapsules. Methods: The chromatographic conditions were as follows: C18 GL column with a mobile phase of acetonitrile:water (92:8 v/v) at flow rate of 1.5 mL/min with detection at 280 nm. The evaluated parameters were specificity, linearity, limits of detection and quantification, precision, accuracy, and robustness. Results: The method was specific and linear (r=0.9982). The limits of detection and quantification were 5.78 μg.mL-1 and 17.54 μg.mL-1, respectively. Suitable accurancy and robustness were obtained. The stability assay showed that pH variation occured after 120 days of storage, and no changes were observed regarding the size and polydispersion parameters. The applicability of the method was evaluated by determining the encapsulation efficiency of the E2 nanocapsules after 120 days of storage. The results showed values >99%. Conclusion: The results demonstrated the applicability of the developed and validated analytical method

Correlation between the care dependence scale of ICU patients and their epidemiological profile

Introducción: La Unidad de Cuidados Intensivos (UCI) es un ambiente donde los pacientes críticos deben ser tratados por un equip multidisciplinario. Es de suma importancia reconocer el perfil clínico epidemiológico para evaluar individualmente los pacientes. Objetivo: Analizar el perfil epidemiológico de los pacientes ingresados en una UCI de un Hospital Universitario y su relación con la escala de Fugulin. Método: Estudio retrospectivo, descriptivo, con enfoque cuantitativo, que realizó un análisis del perfil epidemiológico, desenlaces y variables asociadas a la morbimortalidad, a través de relatos de pacientes hospitalizados de marzo a agosto de 2020. Resultados: Se observó que la mayoría de estos pacientes hospitalizados era del sexo masculino, remitidos por el Servicio de Atención Médica de Urgencias, se les diagnosticó con mayor frecuencia: insuficiencia respiratoria aguda, sepsis e insuficiencia renal aguda, con un desenlace prevalente en muertes, habiéndose observado correlación de la escala de Fugulin con la mortalidad y puntuaciones de gravedad de estos pacientes. Conclusión: Ante la complejidad en el cuidado del paciente crítico, el estudio demuestra que la escala de Fugulin puede ser una alternativa en la práctica clínica, relacionando la necesidad de cuidado con la gravedad y mortalidad de los pacientes en uma UCI.Introduction: The Intensive Care Unit (ICU) is an environment where critical patients must be treated by a multidisciplinary team. What becomes extremely important to recognize the clinical epidemiological profile to evaluate individually. Objective: To analyze the epidemiological profile of patients admitted to an ICU of a University Hospital and its relationship with the Fugulin scale. Method: This is a retrospective, descriptive study with a quantitative approach, which carried out an analysis of the epidemiological profile, outcomes and variables associated with morbidity and mortality, through reports of patients hospitalized from March to August 2020. Results: It was observed that most of these hospitalized patients were male, referred by the Emergency Medical Care Service, were diagnosed more frequently: acute respiratory failure, sepsis and acute renal failure, with a prevalent outcome in deaths, having been observed correlation of the Fugulin scale with the mortality and severity scores of these patients. Conclusion: In view of the complexity in the care of critically ill patients, the study demonstrates that the Fugulin scale can be an alternative in clinical practice, relating the need for care with severity and mortality to patients in an ICU

Correlación entre la Escala de dependencia de cuidados de pacientes ingresados en UCI y su perfil epidemiológico

Introduction: The Intensive Care Unit (ICU) is an environment where critical patients must be treated by a multidisciplinary team. What becomes extremely important to recognize the clinical epidemiological profile to evaluate individually. Objective: To analyze the epidemiological profile of patients admitted to an ICU of a University Hospital and its relationship with the Fugulin scale. Method: This is a retrospective, descriptive study with a quantitative approach, which carried out an analysis of the epidemiological profile, outcomes and variables associated with morbidity and mortality, through reports of patients hospitalized from March to August 2020. Results: It was observed that most of these hospitalized patients were male, referred by the Emergency Medical Care Service, were diagnosed more frequently: acute respiratory failure, sepsis and acute renal failure, with a prevalent outcome in deaths, having been observed correlation of the Fugulin scale with the mortality and severity scores of these patients. Conclusion: In view of the complexity in the care of critically ill patients, the study demonstrates that the Fugulin scale can be an alternative in clinical practice, relating the need for care with severity and mortality to patients in an ICU.Introducción: La Unidad de Cuidados Intensivos (UCI) es un ambiente donde los pacientes críticos deben ser tratados por un equip multidisciplinario. Es de suma importancia reconocer el perfil clínico epidemiológico para evaluar individualmente los pacientes. Objetivo: Analizar el perfil epidemiológico de los pacientes ingresados en una UCI de un Hospital Universitario y su relación con la escala de Fugulin. Método: Estudio retrospectivo, descriptivo, con enfoque cuantitativo, que realizó un análisis del perfil epidemiológico, desenlaces y variables asociadas a la morbimortalidad, a través de relatos de pacientes hospitalizados de marzo a agosto de 2020. Resultados: Se observó que la mayoría de estos pacientes hospitalizados era del sexo masculino, remitidos por el Servicio de Atención Médica de Urgencias, se les diagnosticó con mayor frecuencia: insuficiencia respiratoria aguda, sepsis e insuficiencia renal aguda, con un desenlace prevalente en muertes, habiéndose observado correlación de la escala de Fugulin con la mortalidad y puntuaciones de gravedad de estos pacientes. Conclusión: Ante la complejidad en el cuidado del paciente crítico, el estudio demuestra que la escala de Fugulin puede ser una alternativa en la práctica clínica, relacionando la necesidad de cuidado con la gravedad y mortalidad de los pacientes en uma UCI

A SIMPLE AND FAST RP-UHPLC-PDA METHOD FOR DETERMINATION OF ISRADIPINE FROM POLYMERIC NANOCAPSULES

A simple and fast analytical method of ultra-high performance liquid chromatography (UHPLC) was developed and validated in order to assay isradipine in poly(ε-caprolactone) (PCL)/polyethylene glycol (PEG) nanocapsules. Experiments were performed by UHPLC on a C18 chromatographic column at 25°C using a mobile phase composed by methanol and water (85:15 v/v) with a flow rate of 0.5 mL.min−1 and UV detection at 327nm for achieving a total run time of 1.5 min. The UHPLC method was validated according to the guidelines set on The International Conference on Harmonisation. It proved to be selective, linear (r = 0.99962), precise (RSD < 4.1%), and accurate (recovery rates between 95.24 and 96.53%) at the range from 10 to 40 µg.mL−1. The performance was robust when slight changes in the flow rate, wavelength of detection, and mobile phase composition were tested. It was successfully applied to quantify isradipine from nanoparticulate polymeric systems, showing high loading efficiency rates, greater than 98.55%. These results provided an experimental basis to use this method for quantifying isradipine with reliable results, besides being very fast, easy to perform and cheaper

Evaluation of cutaneous wound healing effect of associated zinc oxide (ZnO) and estrogen (E2) nanoparticles / Avaliação da cicatrização de feridas cutâneas tratadas com a associação de nanopartículas de óxido de zinco (ZnO) e estrógeno (E2)

O objetivo deste estudo foi avaliar o efeito de nanopartículas de estrógeno (EN) e óxido de zinco (ZnON) em feridas cutâneas de ratas ooforectomizadas. O ensaio animal foi realizado com 90 ratas Wistar divididas igualmente em 6 grupos: controle ooforectomizado (CO), grupo tratado com nanopartículas sem o fármaco (PCLN), grupos tratados com ZnOC 2.5 + EC 0.06; ZnON 2.5 + EN 0.06; ZnOC 5.0 + EC 0.12 and ZnON 5.0 + EN 0.12. Foi realizada análise da retração da lesão, parâmetros hematológicos, quantificação de infiltrado inflamatório e fibras colágeno tipo I e III. Não foi possível observar diferença estatística na taxa de retração da lesão, no entanto a análise hematológica de linfócitos e neutrófilos apresentou baixo efeito sistêmico nos grupos tratados com as nanopartículas associadas. A quantificação de células inflamatórias mostrou que houve uma melhor modulação da inflamação no grupo tratado com ZnON 5.0 + EN 0.12. Esses resultados mostraram que o tratamento tópico com a associação de nanopartículas de estrógeno e óxido de zinco pode modular o processo inflamatório, resultando em uma melhor progressão da cicatrização

Surface morphology and in vitro leachability of soft liners modified by the incorporation of antifungals for denture stomatitis treatment

Objective: To evaluate the surface morphology and in vitro leachability of temporary soft linings modified by the incorporation of antifungals in minimum inhibitory concentrations (MIC) for Candida albicans biofilm. Methodology: Specimens of soft lining materials Softone and Trusoft were made without (control) or with the addition of nystatin (Ny), miconazole (Mc), ketoconazole (Ke), chlorhexidine diacetate (Chx), or itraconazole (It) at their MIC for C. albicans biofilm. The surface analyses were performed using Confocal laser scanning microscopy after 24 h, 7 days, or 14 days of immersion in distilled water at 37ºC. In vitro leachability of Chx or Ny from the modified materials was also measured using Ultraviolet visible spectroscopy for up to 14 days of immersion in distilled water at 37ºC. Data (μg/mL) were submitted to ANOVA 1-factor/Bonferroni (α=0.05). Results: Softone had a more irregular surface than Trusoft. Morphological changes were noted in both materials with increasing immersion time, particularly, in those containing drugs. Groups containing Chx and It presented extremely porous and irregular surfaces. Both materials had biexponential release kinetics. Softone leached a higher concentration of the antifungals than Trusoft (p=0.004), and chlorhexidine was released at a higher concentration than nystatin (p<0.001). Conclusions: The surface of the soft lining materials changed more significantly with the addition of Chx or It. Softone released a higher concentration of drugs than Trusoft did, guiding the future treatment of denture stomatitis