Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The present study checks the preventive and therapeutic effect of the liquid emulsion of a mixture of essential oils of oregano (Origanum vulgare) and rosemary (Rosmarinus officinalis) in the control of the neonatal diarrhea in sucking piglets of same (therapeutic) diarrheic litter and without clinical demonstration of diarrhea (preventively), born of females with varied orders of childbirth, these animals were divided in two groups, being that the first group of the essential therapeutic oils (GOET) was divided in three treatments: the treatment controls that contained 63 piglets that received 2mL kg 1 of physiologic solution; the treatment with essential oils of oregano (60 % of carvacrol and 15 % thymol and 2 % of essential oil of rosemary) with 148 piglets, which received the liquid emulsion to 5 % in the dose of 2mL kg 1 and the treatment with antibiotic (enrofloxacin) that contained 124 piglets that received to dosage of 1mL 5kg-1 of lively weight of the antimicrobial one. The second group of the preventive essential oils (GOEP) was divided in two treatments: the control treatment that contained 661 piglets that received 1 mL kg 1 physiologic solution and the treatment of essential oils of oregano (60 % of carvacrol and 15 % thymol) and 2 % of essential oil of rosemary that had 672 animals that received the liquid emulsion to 5 % in the dose of 1mL kg 1. The treatments of the group GOET were administered orally during three consecutive days from the confirmation of the demonstration of the clinical signs of diarrhea and for the treatments of the GOEP were administered orally the target medicines from the confirmation of non demonstration of the clinical signs of diarrhea, during the first and third thing days of pig s life. The experimental delineation was of blocks at random for the GOET and for GOEP was composed by two simple treatments, being that for the parameters cure rate and mortality rate of piglets (rates analyzed inside both groups), each animal represented a repetition. The results allowed to conclude that the antibiotic enrofloxacin can be a total substituted by mixture of essential oils for the GOET, on the other hand for the GOEP the results define that the mixture of essential oils with rosemary and oregano s base do not prevent the neonatal diarrheas of the piglets. / O presente estudo avalia a substituição do antibiótico enrofloxacina e o efeito preventivo e terapêutico da emulsão líquida de uma mescla de óleos essenciais de orégano (Origanum vulgare) e alecrim (Rosmarinus officinalis) no controle da diarréia neonatal em leitões lactantes. Estes são provenientes de leitegadas diarréicas (terapêutico) ou sem manifestação clinica de diarréias (preventivo), nascidos de fêmeas com ordens de partos variadas. Estes animais foram divididos em dois grupos, sendo que o primeiro grupo dos óleos essenciais terapêuticos (GOET) foi dividido em três tratamentos: o tratamento controle que continha 63 leitões que receberam 2 mL kg-1 de solução fisiológica; o tratamento com óleos essenciais de orégano (60% de carvacrol e 15% timol e 2% de óleo essencial de alecrim) com 148 leitões, os quais receberam a emulsão líquida a 5% na dose de 2mL kg-1 e o tratamento com antibiótico enrofloxacina que continha 124 leitões que receberam a dosagem de 1 mL 5kg-1 de peso vivo do antimicrobiano. O segundo grupo dos óleos essenciais preventivos (GOEP) foi dividido em dois tratamentos: o tratamento controle que continha 661 leitões que receberam 1 mL kg-1 solução fisiológica e o tratamento com óleos essenciais de orégano (60% de carvacrol e 15% timol) e 2% de óleo essencial de alecrim que era composto por 672 animais que receberam a emulsão líquida a 5% na dose de 1 mL kg-1.. Os tratamentos do grupo GOET foram administrados via oral durante três dias consecutivos a partir da confirmação da manifestação dos sinais clínicos de diarréia e para os tratamentos do GOEP foram administrados oralmente os medicamentos alvos a partir da confirmação da não manifestação dos sinais clínicos de diarréia, durante o primeiro e terceiro dias de vida do suíno. O delineamento experimental foi de blocos ao acaso para o GOET e para GOEP foi composto por dois tratamentos simples, sendo que para os parâmetros de índice de cura e índice de mortalidade de leitões (índices analisados dentro dos dois grupos), cada animal representou uma repetição. Os resultados permitiram concluir que o antibiótico enrofloxacina pode ser totalmente substituído por mescla de óleos essenciais para o GOET, já para o GOEP os resultados definem que a mescla de óleos essenciais a base de orégano e alecrim não previnem as diarréias neonatais dos leitões.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Rosmarinus officinalis L., popularly known as rosemary, is a medicinal and aromatic species used in the cosmetics and food processing industries, with interest for the herbal industry. This study aimed to determine the biomass production and volatile oil during the growth of rosemary plants cultivated under different salinity (nutritious solution concentrations) and different nitrogen (N) concentrations, and to evaluate the effect of the extracts and volatile oil of the species on the Allium cepa L. cell cycle and genetic material, as well as knowing the phenolic compounds present in the extracts and the chemical composition of volatile oil. The work was divided into two stages: the first aimed evaluating the production of biomass and volatile oil during the rosemary plants growth cultivated under different nutritious solution concentrations, as well as the effect of oil and extracts on the A. cepa cell cycle and genetic material, besides determining the chemical composition of the oil and the phenolic compounds present in the extracts; the second stage aimed evaluating biomass and volatile oil production during the rosemary plants growth cultivated under different N concentrations, as well as the effect of oil and the extracts on the A. cepa cell cycle and genetic material, and determining the chemical composition of the oil and the phenolic compounds present in the extracts as well. The rosemary leaves were collected and analyzed for dry matter, volatile oil extraction and preparation of aqueous extracts by infusion at 5, 20, and 50 g.L-1 concentrations for plants cultivated under different salinities, and 5 and 50 g.L-1 concentrations for plants cultivated at different N concentrations. The volatile oil was analyzed by Gas Chromatography to knowledge of its constitution, and the aqueous extracts were analyzed by High Performance Liquid Chromatography for phenolic compounds determination. The oil and extracts effect was evaluated on the A. cepa cell cycle and genetic material. The completely randomized design was used, and the biomass production and volatile oil yield data were subjected to analysis of variance with polynomial regression and the remaining data were compared by the Scott-Knott test at the 5% level of probability. The nutritious solution concentration, the N concentration and the plant age interfere into the biomass production, and consequently into the rosemary volatile oil yield. However, do not alter the oil and extracts effect on the A. cepa cell cycle and genetic material. The volatile oil and extracts inhibited cell division in all concentrations, and this effect is dose-dependent. In addition, extracts did not show genotoxicity, in spite of oil concentration of 10% being genotoxic. The majority compounds camphor, 1.8 cineole, α-pinene, β-myrcene and verbenone, beyond of β-pinene compounds only controlling the oil crop plants in the concentrations of 8:05, 13:05 and 15:55 mmol.L-1. The phenolic compounds present in the extracts were rosmarinic acid, chlorogenic acid, caffeic acid, carnosic acid, kaempferol, quercetin and rutin. / Rosmarinus officinalis L., popularmente conhecida como alecrim, é uma espécie medicinal e aromática utilizada na indústria de cosméticos e agroalimentícia, com interesse para a indústria de fitoterápicos. Objetivou-se determinar a produção de fitomassa, óleo volátil, composição química do óleo e compostos fenólicos presentes nos extratos, durante o crescimento de plantas de alecrim cultivadas com diferentes salinidades (concentrações de solução nutritiva) e diferentes doses de nitrogênio (N), além de avaliar o efeito dos extratos e óleo volátil da espécie sobre o ciclo celular e material genético de Allium cepa L.. Foram coletadas as folhas de alecrim para determinação da fitomassa, extração de óleo volátil e preparo de extratos aquosos por infusão. Avaliou-se a produção de fitomassa e óleo volátil durante o crescimento de plantas de alecrim cultivadas com diferentes salinidades (1.0, 2.0, 3.0, 4.0 e 5.0 dS.m-1) e em diferentes doses de N (5.55, 8.05, 10.55, 13.05 e 15.55 mmol.L-1), além de avaliar o efeito do óleo (3 e 10%) e dos extratos (nas concentrações de 5, 20, e 50 g L-1 para as plantas cultivadas em diferentes salinidades e nas concentrações de 5 e 50 g L-1 a partir das plantas cultivadas em diferentes doses de N), sobre o ciclo celular e material genético de A. cepa. O óleo volátil foi submetido à análise por Cromatografia Gasosa para conhecimento de sua constituição, e os extratos aquosos à análise por Cromatografia Líquida de Alta Eficiência para a determinação dos compostos fenólicos. As plantas cultivadas em diferentes salinidades foram coletadas aos 100 e aos 160 dias após o plantio (DAP) e as cultivadas em diferentes doses de N aos 90, 180, 270 e 365 DAP. Foi utilizado o delineamento experimental inteiramente casualizado, os dados de produção de fitomassa e rendimento de óleo volátil foram submetidos à análise de variância com regressão polinomial e os demais dados foram comparados pelo teste Scott-Knott (p<0,05). A salinidade, a dose de N e a idade fisiológica da planta interferem na produção de fitomassa e consequentemente no rendimento de óleo volátil de alecrim e não alteram o efeito do óleo e dos extratos sobre o ciclo celular e material genético de A. cepa. O óleo volátil e os extratos inibiram a divisão celular em todas as concentrações estudadas e esse efeito é dose dependente, mas sem apresentar genotoxicidade, exceto na concentração de 10% quando o óleo foi genotóxico. Os compostos majoritários do óleo foram cânfora, 1.8 cienol, α-pineno, β-myrceno e verbenona, além dos compostos β-pineno majoritários apenas no óleo de plantas cultivadas nas concentrações de 8.05, 13.05 e 15.55 mmol.L-1. Os compostos fenólicos presentes nos extratos foram ácido rosmarínico, ácido clorogênico, ácido cafeico, ácido carnósico, canferol, quercetina e rutina.

Made available in DSpace on 2016-05-02T13:55:34Z (GMT). No. of bitstreams: 1 Dissertacao Lucimari Romana Dipe de Faria.pdf: 314985 bytes, checksum: e1af9bd84f61c9e4f4ae004b8e226bcb (MD5) Previous issue date: 2005-03-15 / Coordenacao de Aperfeicoamento de Pessoal de Nïvel Superior / The Rosmarinus officinalis L (rosemary) is widely used in popular medicine The objective of this paper was o study the essential oil of this species (ROEO) on inflammatory and hyperalgesic models in vivo in order to detect the possible involvement of its active principles on the inflammatory and algesic responses and analyze its ulcerogenic potential The following experimental models were used rat paw edema granuloma test vascular permeability ear erythema by croton oil writhing test in mouse formalin test hot plate and gastric ulcer by stress The yield of the essential oil extracted by hydrodestilation of the aerial parts from Rosmarinus officinalis was 1.4% With the oral administration in rats and mice of the doses (50 100 200 500 1000 2000 and 5000 mg/kg) it was not possible to determine the median lethal dose (LD50) With the oral doses of 50 100 200 and 300 mg/kg on the rat paw edema the ROEO produced dose-dependent effect of which the lineal correlation coefficient was r = 0.9986 (y = -13.93 + 0.30) and the median effective dose (ED50) was 212 mg/kg In the writhing test in mice the oral doses of ROEO 10 50 100 200 300 and 400 mg/kg produced dose-dependent effect with a correlation coefficient of r = 0.93095 (y = 30.54 + 0.073) and ED50 of 261 mg/kg The daily administration of ROEO (300 mg/kg, orally) for 6 days inhibited the granulomatous tissue formation in 59% (p <0.001 Student t test) In the ear erythema by croton oil the ROEO in the oral dose of 300 mg/kg provoked the significant inhibitory effect of 77% (p <0.05) In the vascular permeability test the oral treatment with ROEO antagonized the response to histamine thus inhibiting the vascular permeability in 50% (p <0.05, Student t test) The oral treatment with ROEO (300 mg/kg) inhibited phase I of the hyperalgesic process by formalin in 40% (p <0.05 Student t test) while phase II was inhibited in 48% In the hot plate test ROEO (300 mg/kg, orally) produced more latency times than basal times the difference however was not significant In the gastric ulcer trial the ROEO-treated group inhibited in 44% the number of gastric lesions by stress when compared with the control group The results suggest that ROEO shows antiinflammatory activity in both acute and chronic inflammatory processes exhibits analgesic activity and provokes little harmful interference on the gastric mucosa / O Rosmarinus officinalis L (alecrim) é muito utilizado na medicina popular Este trabalho teve como objetivo estudar o óleo essencial desta espécie (OERO) em modelos inflamatórios e hiperalgésicos in vivo a fim de detectar o possível envolvimento de seus princípios ativos na resposta inflamatória algésica e analisar o potencial ulcerogênico Para tanto foram utilizados os seguintes modelos experimentais edema de pata de rato teste do granuloma permeabilidade vascular eritema de orelha por óleo de croton teste de contorções em camundongos teste da formalina placa quente e úlcera gástrica por estresse O rendimento do óleo essencial extraído por hidrodestilação das partes aéreas de Rosmarinus officinalis foi de 1,4% Em ratos e camundongos com a administração das doses (50 100 200 500 1000 2000 e 5000 mg/kg) por via oral (v o) não foi possível determinar a dose letal média (DL50) Com as doses de 50 100 200 e 300 mg/kg v o sobre o edema de pata de rato o OERO produziu efeito dose-dependente cujo coeficiente de correlação linear obtido foi r = 0.9986 (y = -13,93 vezes + 0,30) e a DE50 determinada foi de 212 mg/kg No teste de contorções em camundongos as doses do OERO de 10 50 100 200 300 e 400 mg/kg v o produziram efeito dose-dependente com coeficiente de correlação de r = 0,93095 (y = 30,54 vezes + 0,073), e DE50 de 261 mg/kg A administração diária de 300 mg/kg v o do OERO durante 6 dias, inibiu a formação do tecido granulomatoso em 59% (p < 0,001, teste t de Student) No eritema de orelha por óleo de croton o OERO na dose de 300 mg/kg v o provocou efeito inibitório significativo de 77% (p < 0.05) No teste de permeabilidade vascular o tratamento por via oral com o OERO antagonizou a resposta à histamina inibindo a permeabilidade vascular em 50% (p < 0.05 teste t de Student) O tratamento com o OERO (300 mg/kg v o) produziu inibição de 40% (p < 0.05, teste t de Student) da fase I do processo hiperalgésico induzido por formalina enquanto a inibição da fase II foi de 48% No teste de placa quente o OERO (300 mg/kg v o) produziu tempos de latência maiores que os tempos basais Entretanto tal diferença não foi significativa No ensaio de úlcera gástrica o grupo tratado com o OERO inibiu em 44% o número de lesões gástricas por estresse quando comparado ao grupo controle A partir dos resultados obtidos sugere-se que o OERO apresenta atividade antiinflamatória sobre os processos inflamatórios agudo e crônico e atividade analgésica Entretanto provoca pouca interferência danosa sobre a mucosa gástrica

Šio darbo tikslas - pagaminti kvapiųjų rozmarinų etanolines ištraukas (tinktūras, skystuosius ekstraktus) ir įvertinti jų kokybę. Darbo uždaviniai:  Remiantis literatūros duomenimis parinkti žaliavos ir ekstrahento santykį bei ištraukų gamybos metodus.  Įvertinti pagamintų ištraukų kokybę, nustatant sauso likučio ir fenolinių junginių kiekį.  Įvertinti technologinių parametrų daromą įtaką veikliųjų medžiagų išsiskyrimui iš žaliavos.  Įvertinti ištraukų antioksidacinį aktyvumą. Bendras fenolinių junginių kiekis rozmarinų tinktūrose ir ekstraktuose nustatytas spektrofotometriniu metodu (spektrofotometras UNICAM Helios α UV, Didžioji Britanija) pagal rozmarino rūgšties standartą. Antioksidacinis aktyvumas nustatytas spektrofotometriškai, panaudojant DPPH• laisvojo radikalo sujungimo metodą. Sausasis rozmarinų etanolinių ištraukų likutis nustatytas drėgnomačiu MLS 50-3HA160. Maišymo įtaka išsiekstrahavusių fenolinių junginių kiekybei nustatyta, naudojant magnetinę maišyklę. Tiriant etanolio koncentracijos įtaką išsiekstrahavusių veikliųjų junginių kiekybei, nustatyta, kad optimalus ekstrahentas yra 50 % etanolis. Tačiau antioksidacinio aktyvumo priklausomybė nuo etanolio koncentracijos yra nežymi. Rezultatai parodė, kad rozmarinų tinktūrų ir skystųjų ekstraktų antioksidacinis aktyvumas nepriklauso nuo vaisto formos. Ištirta, kad sauso likučio ir fenolinių junginių kiekis yra didesni skystuose ekstraktuose lyginant su rozmarinų tinktūromis. Nustatyta, kad brinkinimo laikas... [toliau žr. visą tekstą] / The aim of this research is to make ethanol extracts (tinctures and liquid extracts) of Rosmarinus officinalis and evaluate their quality. The tasks of this research are:  According to the literature, to select proportion of the substance and extractive solvent and select the method of production of ethanol extracts.  To evaluate the quality of produced extracts by fixing the dry residual and phenol compounds.  To evaluate an impact of technologic parameters for the quantity of active compounds.  To evaluate an antioxidative activity of ethanol extracts. The total amount of phenol compounds in tinctures and extracts of Rosmarinus officinalis is estimated spectrophotometrically (spektrophotometer UNICAM Helios α UV, Great Britain). Rosmarinic acid was used as a standart. Antioxidative activity (AA) was determined spectrophotometrically by free radical DPPH• method. The dry residual of ethanol extracts was determined by hygrometer MLS 50-3HA160. The impact of stirring on the quantity of seperated phenol compounds was evaluated by magnetic stirrer. Searching the impact of ethanol concentration on the quantity of seperated active compounds, it‘s evaluated that optimal extractive solvent is 50 % ethanol. However AA dependence on ethanol concentration is low. The findings confirm that AA of tinctures and extracts of Rosmarinus officinalis doesn‘t depend on drug form. It‘s evaluated that the quantity of dry residual and phenol compounds in liquid extracts is more than in... [to full text]

Made available in DSpace on 2014-06-11T19:23:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-05-06Bitstream added on 2014-06-13T19:09:25Z : No. of bitstreams: 1 cordeiro_chg_me_arafcf.pdf: 668360 bytes, checksum: 6006843241d7fb47bcd17a5594004e3c (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / A utilização de produtos naturais na medicina popular é milenar e persiste até os dias atuais. Sua abrangência é vasta e engloba fins variados, inclusive em relação à saúde bucal. O controle do biofilme bacteriano dentro das diversas especialidades odontológicas é de grande importância, pois aponta para a prevenção de várias afecções bucais. Quando a remoção mecânica do biofilme é deficiente, podem ocorrer lesões de tecido duro (cáries) e/ou mole (doenças periodontais). A eficácia de plantas medicinais, tanto em dentifrícios como em enxagüatórios bucais, tem sido investigada. Os resultados têm sugerido que os componentes de origem vegetal podem ser empregados como apoio à terapia das doenças periodontais e como profilaxia de rotina. Este trabalho teve como objetivos avaliar a ação biológica de formulações para higiene bucal, contendo, em associação, extratos hidroalcoólicos das espécies vegetais Nasturtium officinale (agrião), Rosmarinus officinalis (alecrim), Tabebuia impetiginosa (ipê roxo), Achillea millefolium (mil-folhas) e Plantago major (tanchagem). Trabalhos anteriores sugerem possível efeito antiinflamatório e alteração da qualidade do biofilme dento-bacteriano, devido as substâncias ativas desses vegetais. Após os estudos de pré-formulação e obtenção do gel dentifrício e do enxagüatório bucal, foram realizadas avaliações da atividade antibacteriana (frente à Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Enterococcus faecalis e Pseudomonas aeruginosa), atividade antiinflamatória e imunológica, para os extratos vegetais e para as formulações. E foram ainda realizados estudos clínicos iniciais, considerando-se os parâmetros odontológicos para a avaliação do Índice de placa, Índice Gengival, Recessão Gengival e... . / The use of natural resources in popular medicine is a millenary practice and it persists until nowadays. This habit is embracing and wide and it conglomerates multiple purposes, included those related to oral health. It’s very important to control bacterial film in many odontologic specialties, because this control helps to prevent several oral diseases. When the mechanical removal of the bacterial film is inefficient, there can occur hard tissue lesions (carious lesions) and/or soft tissue lesions (periodontal diseases) The effectiveness of medicinal herbs in dentifrices, as well as in mouthwashs have been investigated. The results have suggested that the herbs components can be utilized as a support in the periodontal diseases therapies and in the routine prophylaxis. It was the purpose of the present study to evaluate the biological action of various formulations to bucal hygiene, containing, in association, hidroalcoholic extracts from the following species: Nasturtium officinale, Rosmarinus officinalis, Tabebuia impetiginosa, Achillea millefolium and Plantago major. Previous works suggested that there are possible anti-inflammatory effects and an alteration of the bacterial dental film quality, due to the active substances of these herbs. After studying a previous formulation and obtaining the dentifrice and the mouthrinse, it was evaluated the bacterial activity (related to Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa), the antiinflammatory and the immunological activities, for the herbs extracts and for the formulations... (Complete abstract, click electronic address below).

Submitted by JONÂTAS RAFAEL DE OLIVEIRA null (jroliveira16@hotmail.com) on 2016-04-06T20:01:18Z No. of bitstreams: 1 #TESE COMPLETA.pdf: 2320841 bytes, checksum: da9a41fcb44d79b7879a708f1c2de87e (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-04-08T16:37:41Z (GMT) No. of bitstreams: 1 oliveira_jr_dr_sjc.pdf: 2320841 bytes, checksum: da9a41fcb44d79b7879a708f1c2de87e (MD5) / Made available in DSpace on 2016-04-08T16:37:41Z (GMT). No. of bitstreams: 1 oliveira_jr_dr_sjc.pdf: 2320841 bytes, checksum: da9a41fcb44d79b7879a708f1c2de87e (MD5) Previous issue date: 2016-02-12 / Não recebi financiamento / No presente estudo foram avaliadas algumas atividades biológicas dos extratos de R. officinalis L. (alecrim) e T. vulgaris L. (tomilho), verificando: I. Atividade antimicrobiana sobre biofilmes monomicrobianos de Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans e Pseudomonas aeruginosa e associações de C. albicans com cada uma destas bactérias, após determinação da concentração inibitória mínima (CIM) e concentração microbicida mínima (CMM) sobre culturas planctônicas destes micro-organismos; II. Viabilidade celular sobre macrófagos de camundongo (RAW 264.7), fibroblastos gengivais humanos (FMM-1), linhagem tumoral de carcinoma mamário (MCF-7) e linhagem tumoral de carcinoma cervical (HeLa); III. Atividade anti-inflamatória sobre RAW 264.7 estimulada por lipopolissacarídeo (LPS) de Escherichia coli; e IV. Atividade genotóxica. Após verificação da capacidade antimicrobiana dos extratos, por método de microdiluição em caldo, as concentrações mais efetivas foram aplicadas nos demais testes. Sobre os biofilmes mono e polimicrobianos, formados por 48 h em poços de placa de microtitulação, a ação dos extratos foi analisada após exposição de 5 min, com quantificação de UFC/mL e analise de sua viabilidade. Da mesma forma, sobre as linhagens celulares (RAW 264.7, FMM-1, MCF-7 e HeLa), cultivadas por 24 h em placas de microtitulação, os extratos foram analisados com os ensaios de MTT, vermelho neutro (VN) e cristal violeta (CV). Foi analisado o efeito anti-inflamatório dos extratos com ensaio imunoenzimático ELISA, pela quantificação de citocinas pró-inflamatórias (TNF-α e IL-1β). A genotoxicidade dos extratos foi verificada pela frequência de micronúcleos (MN) formados em 1000 células. Os resultados foram analisados por T-Test ou ANOVA e Tukey Test (p ≤ 0,05). Em relação ao extrato de R. officinalis L. foi observada redução significativa de todos os biofilmes após exposição de 5 min, bem como, redução de sua viabilidade. Quanto às células, foi observado viabilidade celular acima de 50% a ≤ 50 mg/mL. O nível de IL-1β não se alterou com as concentrações do extrato, entretanto quando RAW 264.7 foi estimulada por LPS, houve inibição da produção desta citocina. Em relação ao nível de TNF-α, houve modulação da síntese desta citocina, sem e com LPS. Foi constatado também ausência de genotoxicidade, uma vez que, a frequência de MN foi semelhante ou menor ao grupo controle. Em relação ao extrato de T. vulgaris L., todos os biofilmes mono e polimicrobianos apresentaram reduções significativas em comparação ao grupo controle. A viabilidade dos biofilmes também foi reduzida. Sobre todas as linhagens celulares foi observada viabilidade celular acima de 50%. Efeito imunomodulador foi observado na síntese de IL-1β e TNF-α. A formação de MN foi semelhante ou inferior ao grupo controle, em todas as linhagens. Com isso, foi concluído que ambos os extratos foram efetivos sobre biofilmes mono e polimicrobianos, proporcionaram viabilidade celular para as culturas acima de 50%, apresentaram efeito imunomodulador e não foram genotóxicos. / This study performed some biological activities of R. officinalis L. (rosemary) and T. vulgaris L. (thyme) extracts, such as: I. Antimicrobial activity on monomicrobial biofilms of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Pseudomonas aeruginosa and C. albicans association with each of these bacteria, after determination of the minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) on planktonic cultures of these microorganisms; II. Cell viability on mouse macrophages (RAW 264.7), human gingival fibroblasts (FMM-1), human breast carcinoma cells (MCF-7) and cervical carcinoma cells (HeLa); III. Anti-inflammatory activity on lipopolysaccharide (LPS)-stimulated RAW 264.7; and IV. Genotoxic activity. After checking the antimicrobial capacity of the extracts by broth microdilution method, the most effective concentrations were applied in the other tests. On mono- and polymicrobial biofilms formed by 48 h in microtitre plate wells, the action of the extracts was analyzed after 5 min exposure, with counting of CFU/mL and analysis of their viability. Likewise, in the cell lines (RAW 264.7, FMM-1, MCF-7 and HeLa cells) cultured for 24 h in microtitre plates, the extracts were analyzed by MTT, neutral red (NR) and crystal violet (CV) assays. By ELISA assay, anti-inflammatory effect of the extracts was analyzed with quantification of proinflammatory cytokines (TNF-α and IL-1β). The genotoxicity of the extracts was verified by the micronuclei frequency (MN) formed in 1000 cells. The results were analyzed by T-Test or ANOVA and Tukey test (p ≤ 0.05). Regarding the R. officinalis L. extract was observed significant reduction of all biofilms after 5 min of exposure, as well as reduction their viability. Additionally, cell viability above 50% to ≤ 50 mg/mL was observed on lineages. The IL-1β level did not change with the concentrations of the extract, but when RAW 264.7 was stimulated by LPS, there was inhibition of the production of this cytokine. In relation to the level of TNF-α, was modulation of the synthesis of this cytokine with and without LPS. It was also observed lack of genotoxicity, since the frequency of MN was similar or less than the control group. Regarding to T. vulgaris L. extract, all mono and polymicrobial showed significant reductions compared to the control group. Their viability was also reduced. On all cell lines was observed cell viability above 50%. Immunomodulatory effect was observed in IL-1β and TNF-α synthesis. The MN formation was similar or lower than the control group in all cell lines. Thus, it was concluded that both extracts were effective on mono- and polymicrobial biofilms, promoted cell viability above 50% for cultures, presented immunomodulatory effect and were non-genotoxic.

Orientadores : Jose Francisco Hofling, Mary Anny Foglio / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-08T15:40:59Z (GMT). No. of bitstreams: 1 Mardegan_RitadeCassia_D.pdf: 1623661 bytes, checksum: 6efd1058d06bb35d4bbde2640edda2e8 (MD5) Previous issue date: 2007 / Resumo: O uso de extratos vegetais com fins medicinais é uma das mais antigas formas de prática medicinal da humanidade. Visto que nos últimos anos, a freqüência das infecções fúngicas sistêmicas, principalmente as oportunistas invasivas, têm crescido drasticamente o objetivo deste estudo foi: 1 - testar a atividade dos extratos (diclorometanico e metanólico) de Rosmarinus officinalis, Mentha piperita, Casearia sylvestris, Arctium lappa, Arrabidaea chica e Tabebuia avellanedae para determinar o potencial antifúngico em cepas padrão de Cândida spp (C. albicans, C. dubliniensis, C. parapsilosis, C. tropicalis e C. krusei) e em amostras clinicas de Candida albicans através da determinação da Concentração Inibitória Mínima (CIM) e; 2 - avaliar a capacidade desses extratos em inibirem a atividade das proteinases produzidas por C. albicans, um dos mais importantes fatores de virulência dessa espécie. Foram incluídos nos testes para analisar a inibição da atividade proteolítica os inibidores de protease (Amprenavir e Ritonavir) e a Pepstatina A. Nossos resultados demonstraram CIMs variadas em relação a sensibilidade de cada cepa. O extrato metanólico de Arrabidaea chica foi o mais efetivo na inibição do crescimente de várias espécies de Candida, inclusive em relação aos isolados clinicos de C. albicans, seguido dos extratos diclorometanicos de Arctium lappa e Mentha piperita. Nos testes de inibição das proteinases de Candida albicans os que se destacaram foram os extratos diclorometanico de Arrabidaea chica, Casearia sylvestris e Mentha piperita. Desta forma concluímos que o extrato metanólico de A. chica e os extratos diclorometanicos de A. lappa, M. piperita e C. sylvestris obtiveram relevante atividade antifúngica in vitro contra uma variedade de espécies de Candida sendo promissores como agente antifúngico. / Abstract: The use of plants as medicinal substances is one of the most old medicine practices of the humanity. In the last years, the frequency of systemic yeast infection, mainly the opportunistic, has been increased. The overall aims of the present study were: 1 - to test the extract activity (dichloromethane and methanolic) from Rosmarinus officinalis, Mentha piperita, Casearia sylvestris, Arctium lappa, Arrabidaea chica and Tabebuia avellanedae to determine the antifungal activity using strains of Candida spp (C. albicans, C. dubliniensis, C. parapsilosis, C. tropicalis e C. krusei) and in clinical isolates of C. albicans by determination of minimum inhibitory concentration (MIC) and 2 - evaluate the capacity of the extracts to inhibit the activity of proteinases produced by C. albicans, one of the most important virulence factor of these specie. It has been included as a control to evaluate the inhibition of proteolitic activity Amprenavir e Ritonavir and Pepstatina A. Our results showed a variety of MIC patterns according to the strain sensibility. The methanolic extract of Arrabidaea chica was the most effective in inhibiting the growth of a variety of Candida species, including the clinical isolates of C. albicans, followed by dichloromethane extract of Arctium lappa and Mentha piperita. In relation of the proteinases inhibitory tests from C. albicans the most effective extracts tested were dichloromethane of Arrabidaea chica, Casearia sylvestris and Mentha piperita. In conclusion the methanolic extract from A. chica and dichloromethane extracts from A. lappa, M. piperita e C. sylvestris have relevant antifungal activity against a range of Candida specie in vitro and are promising antifungal agents. / Doutorado / Microbiologia e Imunologia / Mestre em Biologia Buco-Dental

Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts.

El estudio del impacto de los eventos extremos de sequía sobre los ecosistemas secos mediterráneos es fundamental para desarrollar herramientas de gestión adaptativa frente al cambio climático. La complejidad en la respuesta de los ecosistemas y las dificultades experimentales para llevar a cabo estudios de campo sobre eventos extremos de sequía, son los principales obstáculos en este campo de investigación. Es necesario desarrollar metodologías que permitan estudiar la relación de los ecosistemas con la disponibilidad hídrica y parametrizar los eventos de sequía extrema para poder resolver las inconsistencias surgidas hasta el momento en este ámbito. En la presente tesis se han estudiado las respuestas de los ecosistemas a distintos niveles ecológicos, desde el sistema suelo planta hasta los procesos ecosistémicos, frente a cambios en la disponibilidad hídrica durante eventos extremos de sequía. Para ello, se han realizado dos experimentos manipulativos de lluvia y se ha analizado un evento de sequía extrema natural en seis comunidades vegetales dominadas por arbustos del Sureste de la Península Ibérica, en la Comunidad Valenciana. Los resultados de estos estudios, mostrados a lo largo de seis capítulos en esta tesis, indican que la sequía extrema, y los cambios en la disponibilidad hídrica que ella produce, afectan a estos ecosistemas a todos los niveles ecológicos estudiados. Además, apuntan a la relevancia del patrón de precipitaciones y de la dinámica de la humedad del suelo como principales responsables de la respuesta de los ecosistemas. La respuesta de los ecosistemas a la sequía extrema fue mayor a niveles ecológicos superiores, siendo la reducción en la tasa de descomposición de hojarasca y la acumulación de combustible seco las principales consecuencias tanto a nivel experimental como natural. La sequía extrema también afectó a la fenología y esfuerzo reproductivo de la vegetación, así como a su crecimiento y procesos productivos. Finalmente se observaron cambios en la dinámica de nutrientes edáficos asociados al aumento de intensidad de la sequía. Estos resultados nos permitirán avanzar en la comprensión de las relaciones autoecológicas de la vegetación. La metodología desarrollada en este estudio ha demostrado simular satisfactoriamente las condiciones y efectos de la sequía extrema, mostrándose por tanto como una herramienta válida para realiza este tipo de estudios. / Tesis financiada por una beca predoctoral FPU-UA (2015). La parte experimental de esta tesis ha sido financiada por los proyectos CASCADE [GA 283068] (CEAM), SURVIVE-2 [CGL2015-69773-C2-2-P] y ALTERACLIM [CGL2015-69773-C2-1-P].

Les objectifs de cette thèse ont été de contribuer à l'évaluation des niveaux de contamination en éléments traces métalliques et métalloïdes (ETMM) des sols du site de l'ancienne fonderie de plomb de l'Escalette (sud-est de Marseille), d'en approcher les processus de transport et transfert dans l'environnement en particulier leurs mobilités dans le sol et leurs transferts aux plantes. La contribution à l'établissement d'une cartographie de la contamination sur un périmètre plus vaste que celui de l'usine (zone des Calanques) a permis de mettre clairement en évidence l'impact de cette dernière. Sur le site industriel, les formes et niveaux de contamination des sols diffèrent d'une zone à l'autre. Les sols formés en surface des crassiers se caractérisent par leurs fortes teneurs en Cu, Zn et Pb, dont les fractions mobilisables sont notables (2 à 30%), l'influence des scories est nette. Les sols prélevés à proximité de la cheminée présentent les plus fortes contaminations en Pb, Sb et As, avec de 10 à 30% de fractions mobilisables. Cette contamination est liée aux émissions anciennes via la cheminée rampante mais aussi à la dégradation de cette dernière. Le potentiel de libération des éléments As et Sb des sols de ces 2 zones conduirait à leurs stockages dans des décharges pour déchets dangereux. Les essais de solubilisation en milieu acide ont mis en évidence des risques pour la santé par ingestion concernant Pb et As. Les zones en proche périphérie de l'usine sont moins impactées mais sont néanmoins considérées comme fortement contaminées. G. alypum et R. officinalis, présentes sur le site, sont tolérantes aux ETMM et peuvent les accumuler. / The overall objective of this thesis was to contribute to the assessment of the level of contamination by trace metals and metalloids (TMM) on the site of Escalette (a former lead smelter in southeastern Marseille). The study of TMM transfer and transport processes on this site, taking into account their mobility in the soil and their transfer to plants, was a specific objective. The mapping of TMM contamination in an extended area around the factory site (on Calanques zone) has clearly highlighted the impact of the former industry. So, on the Escalette site, forms and levels of soil contamination differ from one area to another. Soils formed on the surface of the slag deposits are characterized by high levels of Cu, Zn and Pb, whose mobilizable fractions are significant (2-30%), showing a clear influence of slag. Soils collected near a chimney creeping uphill the slope, present the highest contamination levels in Pb, Sb and As, with from 10 to 30 % of mobilizable fractions. This contamination is linked to emissions through the damaged chimney but also to the degradation of this latter. The potential release of As and Sb from the soils of these two areas requires that these soils must be disposed of in landfills for hazardous wastes and their solubilization in acid medium have highlighted the health risks by ingestion for Pb and As. On areas in near periphery of the smelter, soils are less affected but must be still considered heavily contaminated. G. alypum and R. officinalis were observed on the site. They are tolerant and can accumulate TMM. Concentrations in roots were significantly correlated with the concentrations in soils, except for Cu.