A cárie dentária é uma doença bucal de alta prevalência e impactante em países em desenvolvimento. É causada pela presença de biofilme dentário rico em bactérias acidogênicas e acidúricas, como Streptococcus mutans e Lactobacillus casei. Neste sentido, a fitoterapia tem sido aplicada na odontologia devido ao seu conhecido efeito antimicrobiano, tendo potencial para prevenir doenças como a cárie dentária. Portanto, o presente estudo tem como objetivo testar o potencial antimicrobiano de extratos bruto e etanólico das folhas de Myracrodruon urundeuva (M. urundeuva.) e Qualea grandiflora (Q. grandiflora) sobre S.mutans e L. casei. Para tal, determinaram-se a Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM). A CIM foi definida como a menor concentração do agente antimicrobiano capaz de inibir 100% o crescimento microbiano (absorbância) em relação aos controles negativos. Para a CBM, alíquotas foram removidas dos poços que não apresentaram nenhuma absorbância (viabilidade, concentrações CIM) e semeadas em placas de ágar BHI, incubadas por 24 h a 37°C em estufa de CO2 5%. A CBM foi determinada considerando a menor concentração dos extratos capaz de impedir o crescimento bacteriano visível. Cepas de S. mutans (ATCC 21175) e L. casei (ATTC 334) foram ativadas em BHI e caldo Rogosa, respectivamente. A CIM foi determinada pela técnica de diluição em microplacas de 96 poços (100 l de extrato + 80 l BHI/Rogosa + 20 l da bactéria diluída em BHI/Rogosa equivalente a 5x105 UFC/mL), as quais foram incubadas por 24 h (S. mutans) e 48 h (L. casei) a 37°C em estufa de CO2 5%. Os extratos de M. urundeuva e Q. grandiflora inicialmente foram diluídos em BHI/Rogosa variando as concentrações entre 2 mg/ml a 0,00012207 mg/ml e os mesmos extratos diluídos em álcool foram avaliados nas concentrações entre 20 mg/ml a 0,00244 mg/ml para S. mutans e L. casei. Não foi possível determinar a CIM e a CBM para os extratos diluídos no BHI/Rogosa. Foram utilizados como controle positivo a clorexidina e como controles negativos BHI/Rogosa com e sem álcool a 5%. As CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e clorexidina contra S. mutans foram 2,5 mg/ml (2,5 mg/ml), 5,0 mg/ml (--) e 0,00468 mg/ml (0,00937 mg/ml), respectivamente. Em relação ao L. casei, as CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e da clorexidina foram 0,156 mg/ml (0,312 mg/ml), 0,156 0,625 mg/ml (0,312 0,625mg/ml) e 0,00468 mg/ml (0,3 mg/ml), respectivamente. Como conclusão, nosso estudo mostrou que L. casei (ATTC 334) é mais susceptível aos extratos que S. mutans (ATCC 21175) e o extrato M. urundeuva apresenta melhor efeito antimicrobiano que a Q. grandiflora em S. mutans (ATCC 21175), porém os dois extratos apresentam efeito similar sobre L. casei (ATTC 334) e ambos foram inferiores à CHX. / Dental caries is an oral disease of high prevalence and impact in developing countries. It is caused by the presence of a dental biofilm rich in acidogenic and aciduric bacteria, such as Streptococcus mutans and Lactobacillus casei. Accordingly, phytotherapy has been applied in dentistry due to its known antimicrobial effect, having potential to prevent diseases such as dental caries. Therefore, the present study aims to test the antimicrobial potential of crude and ethanolic extracts of Myracrodruon urundeuva (M. urundeuva) and Qualea grandiflora (Q. grandiflora) leaves on S. mutans and L. casei. For this, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. MIC was defined as the lowest concentration of the antimicrobial agent capable of inhibiting 100% the microbial growth in comparison to the negative controls. For MBC, aliquots were removed from the wells that did not show any absorbance (viability, concentrations than MIC) and seeded on BHI agar plates, incubated for 24 h at 37°C and 5% CO2. The MBC was determined considering the lowest concentration of extracts capable of preventing visible bacterial growth. Strains of S. mutans (ATCC 21175) and L. casei (ATTC 334) were activated in BHI and Rogosa broth. MIC was determined by the dilution technique in 96-wells microplates (100 l of extract + 80 l BHI/ Rogosa + 20 l of bacterium diluted in BHI/Rogosa equivalent to 5x105 CFU/ml), which were incubated for 24 h (S. mutans) and 48 h (L. casei) at 37°C and 5% CO2. The extracts were firstly diluted in BHI/Rogosa varying the concentrations between 2 mg/mL and 0.00012207 mg/ml; the same extracts were also diluted in alcohol at concentrations ranging from 20 mg/ml to 0.00244 mg/ml and both tested against S. mutans and L. casei. It was not possible to determine the MIC and MBC for the extracts diluted in BHI/Rogosa. Chlorhexidine was used as positive control, while BHI/Rogosa with or without 5% alcohol were used as negative controls. The MICs (MBCs) of M. urundeuva and Q. grandiflora diluted in alcohol, and clorexidine against S. mutans were 2.5 mg/ml (2.5 mg/ml), 5.0 mg/ml (--) and 0.00468 mg/ml (0.00937 mg/ml), respectively. In respect to L. casei, the MICs (MBCs) of M. urundeuva and Q. grandiflora, diluted in alcohol, and chlorhexidine were 0.156 mg/ml (0.312 mg/ml), 0.156 0.625 mg/ml (0.312 0.625 mg/ml) and 0.00468 mg/ml (0.3 mg/ml), respectively. In conclusion, our study showed that L. casei (ATTC 334) is more susceptible than S. mutans (ATCC 21175) to the extracts and the extract of M. urundeuva has a better antimicrobial effect than Q. grandiflora against S. mutans (ATCC 21175), but both extracts have similar effect on L. casei (ATTC 334) and they were inferior to CHX.

A mastite bovina é considerada a doença que causa os maiores prejuízos à produção leiteira, reduzindo em quantidade e qualidade o leite e os derivados lácteos. Especialmente na mastite clínica, ocorre aumento no risco de resíduos de antimicrobianos no leite, portanto além do prejuízo diretamente relacionado ao processo inflamatório, acrescenta-se o custo com medicamentos, aumento do labor da mão-de-obra e tempo de descarte do leite após tratamento, até a total eliminação os resíduos de antibióticos utilizados. A expansão dos sistemas de produção pecuária orgânica aumenta a necessidade de utilizar métodos diferentes daqueles conhecidos convencionalmente.A homeopatia é uma terapêutica que tem sido cada vez mais utilizada em animais de produção com resultados bastante satisfatórios. No Brasil a produção orgânica é regulamentada pela Lei No10.831 de 23 de dezembro de 2003 e lá há orientação para o uso de medicamentos homeopáticos, assim como fitoterapia e acupuntura na terapêutica animal, em lugar dos medicamentos convencionais, que têm seu uso muito restrito e em certos casos proibido. Para realização deste trabalho foi feita uma inoculação intramamária experimental com estirpes de Staphylococcus aureus em 36 quartos mamários de 18 vacas mestiças de Holandesas e Gir pertencentes ao Campo Experimental de Coronel Pacheco/MG da Embrapa/CNPGL, com o objetivo de comparar o tratamento de animais acometidos com a utilização de medicamentos homeopáticos (Phytolacca decandra 6CH, Calcarea carbonica 6CH e Silicea terra 6CH) e com antibiótico (Cefoperazone Sódico), usando como parâmetros para este estudo: os sinais clínicos, a prova de CMT, as contagens de células somáticas , tanto eletrônicas quanto ópticas e culturas microbiológicas, além da avaliação do custo dos dois tratamentos. No presente estudo não houve diferença estatisticamente significante quanto à intensidade do processo inflamatório avaliados pelo CMT e contagens de células somáticas, quanto ao número de unidades formadoras de colônias isoladas do leite das glândulas mamárias inoculadas e entre período de convalescença dos dois tratamentos. O custo de aquisição dos medicamentos para o tratamento de mastite aguda utilizando homeopatia foi muito inferior ao mesmo tratamento realizado com antibiótico intramamário / The bovine mastitis is considered the illness that causes most losses in milk production, reducing the quantity and quality of milk and dairy products. Especially in clinical mastitis there is an increase on the risk of antimicrobials in milk. Therefore, beyond the losses directly related with the inflammatory process, there is also the expense with drugs, an increase in the animal handling and a discard of milk after the antibiotics treatment, up until the totally elimination of the drug residues. The expansion of organic animal production systems increases the necessity of using different methods of those already conventionally known. Homeopathy is a therapeutically method that has been more used in livestock with satisfactory results. In Brazil, organic production is regulated by the law number 10.831 of December 23rd of 2003, where there is an orientation for using homeopathic medicines as well as phytotherapy and acupuncture in animal health instead of conventional medicines which are restricted in many cases and sometimes even forbidden. In order to develop this research it was done an experimentally intramammary inoculation with a strain of Staphylococcus aureus in 36 mammary gland quarters of 18 cows cross-bred with Holstein and Gir from the Experimental Fields of Coronel Pacheco/MG of Embrapa/CNPGL. The purpose was to compare the treatment of the inoculated animals with homeopathy (Phytolacca decandra 6CH, Calcarea carbonica 6CH and Silicea terra 6CH) and antibiotic (Sodic Cefoperazone). The parameter used for this study were clinical signs, CMT, counting of somatic cells electronically in as much as using the optical method and microbiological cultures. It was also estimated the costs of both treatments. At the present study there was no statistical difference in intensity of the inflammatory process evaluated by the CMT and somatic cell counts, by the number of CFU isolated from milk of the inoculated mammary glands and between the times of convalescence in both treatments. The cost of acquisition of medicines for the treatment of acute mastitis using homeopathy was very lower when compared with the same treatment done with intramammary antibiotics

A Cynara scolymus, popularmente conhecida como alcachofra, é uma planta hepatoprotetora com função colerética e colagoga. O extrato de C.scolymus foi recentemente liberado para a confecção de medicamentos fitoterápicos sem a necessidade de testes toxicológicos específicos, por ser uma planta utilizada há muito tempo e ter eficácia comprovada. Devido à falta de estudos sobre os efeitos dessa planta e por ela ser muito utilizada, o presente estudo visou avaliar os possíveis efeitos imunotóxicos, teratogênicos e de toxicidade geral do extrato seco da Cynara scolymus. O extrato seco da planta foi administrado a ratos Wistar adultos, machos e fêmeas, nas doses de 1, 2 e 4g/Kg, por gavagem, durante 28 dias, de acordo com o protocolo de testes de toxicidade da OECD Organization for Economic Co-operation and Development, para avaliação de toxicidade geral e imunotoxicidade. Foi avaliado o ganho de peso, consumo de alimento e água, os órgãos linfóides e a fenotipagem de suas células; a atividade de fagócitos, as respostas imunes humoral e celular e a atividade proliferativa de linfócitos também foram avaliados. No estudo da teratogenicidade, ratas prenhes receberam as mesmas doses do extrato seco de C.scolymus do 6° ao 19° dia de gestação. Avaliou-se o tamanho, peso, viabilidade e malformações externas dos fetos, bem como procedeu-se à análise óssea e visceral. O presente estudo revelou que o extrato de alcachofra não promoveu toxicidade geral nos ratos adultos tratados subagudamente; por outro lado, o extrato seco de C.scolymus promoveu aumento no peso relativo e celularidade do baço, aumento no peso relativo dos rins, diminuição na celularidade do timo e diminuição no DTH Delayed-type hypersensivity . Apesar disso a análise destes dados em conjunto não permite sugerir efeito imunotóxico dessa planta. O estudo da teratogenicidade mostrou que fêmeas gestantes tratadas com a C.scolymus apresentaram diminuição no ganho de peso; além disso, foi observada diminuição do peso uterino e fetal, menor comprimento fetal e queda do número de fetos vivos por ninhada. Assim, pode-se sugerir que embora a alcachofra não apresente toxicidade geral, nem efeito imunotóxico, não deve ser utilizado durante a gestação, tendo em vista que foi observada toxicidade fetal. / Cynara scolymus, commonly known as artichoke, is popular used as hepatoprotectant, choleretic and colagogue agent. The C.scolymus extrat was recently approved by Brazilian regulatory agency, for production of phytotherapeutic drugs, without toxicological tests, because it has been considered both harmless and effective. Due to lack of toxicological studies and its widely usage, the aim of this study is evaluate the immune system and the teratogenic effects as well as the general toxicity of C. scolymus extracts. Therefore, the extract was administered to adult male and female Wistar rats by gavage at 1, 2 and 4g/Kg doses for 28 days, according to the OECD (Organization for Economic Co-operation and Development) protocols. Body weight gain, food and water consumption, lymphoid organs and their cell phenotype were evaluated as well as the phagocyte analysis, humoural and cellular immune responses, and lymphocyte proliferation. In teratogenic study, pregnant rats received the same doses of C.scolymus extract from the 6th to the 19th gestational day. Fetuses size, weight, viability and external malformations were evaluated and subjected to bone and visceral analysis. The present study showed that C. scolymus extracts did not promote sub-acute toxicity in adult rats; however, the relative spleen weight and it cellularity were increased just as the kidney weight, but the thymus cellularity and DTH (Delayed-type hypersensivity) decreased. None of those results represent immunotoxic effect. The teratogenic study showed a decrease of body weight gain, uterus weight, fetal weight and number of fetuses, in practically all doses of the extract. Thus, these results showed that the C. scolymus extract is not toxic or immunotoxic but should not be used in pregnancy, due to fetal toxicity.

Orientador: José Anchieta Gomes Neto / Banca: Mercedes de Moraes / Banca: Edenir Rodrigues Pereira Filho / Banca: Márcia Andreia Mesquita Silva da Veiga / Banca: Cassiana Seimi Nomura / Resumo: O uso de fitoterápicos tem crescido consideravelmente nos últimos anos. Entretanto, o consumo de produtos derivados de plantas medicinais não está isento de riscos. Além do princípio ativo algumas espécies vegetais podem apresentar em sua composição química substâncias tóxicas (orgânicas ou inorgânicas) que podem causar sérios danos à saúde quando ingeridas. Neste sentido, o presente trabalho teve como objetivo o desenvolvimento de métodos analíticos visando a determinação de As, Cd, Co, Pb, Se e Sn em plantas medicinais empregando a espectrometria de absorção atômica de fonte contínua com alta resolução em forno de grafite com amostragem direta de sólidos (SS HR-CS GFAAS). O emprego de modificadores químicos permitiu calibração em meio aquoso para determinação dos analitos. A utilização de uma etapa de pirólise extra assistida por ar a 600 °C permitiu eliminar resíduos carbonáceos e eliminar os sinais de fundo. Para determinação de As, Co, Se e Sn foram observadas interferências espectrais causadas por moléculas diatômicas de PO e SiO. Nestes casos, o emprego do algoritmo dos mínimos quadrados (LSBC) corrigiu o fundo; entretanto, para a determinação de As observou-se interferências espectrais provocadas por linhas atômicas de Al que somente foram corrigidas utilizando algoritmo de correção de fundo interativa (IBC). Os limites de detecção foram 28,7 ng g-1 (As), 0,15 ng g-1 (Cd), 3,2 ng g-1 (Co), 7,3 ng g-1 (Pb), 40,1 ng g-1 (Se) e 24,5 ng g-1 (Sn). A exatidão dos métodos foi avaliada através da análise de materiais de referência certificados de plantas, cujos valores foram concordantes com os certificados ao nível de 95 % (test t). Os estudos de homogeneidade foram conduzidos visando determinar as massas mínimas e a homogeneidade após moagem com moinho criogênico e moinho de facas. A melhor precisão e exatidão foi obtida a partir da análise de massas maiores que 0,2 mg de amostra... / Abstract: Phytotherapy has increased considerably during the last years. However, besides active compounds, products derived from medicinal plants may contain hazardous organic compounds or toxic metals. Thus, the goal of this work was to develop methods to determine As, Cd, Co, Pb, Se and Sn in medicinal plants by high-resolution continuum source atomic absorption spectrometry using solid sampling. The use of chemical modifiers allowed the determination of the analytes of interest using calibration with aqueous standard solutions. An additional air-assisted pyrolysis step at 600 °C was useful to remove carbonaceous residues and eliminate background signal. Spectral interferences due to structured background caused by diatomic molecules (PO and SiO) were observed on the analytical lines of As, Cd, Co and Sn. In this case, interferences were removed using least squares background correction algorithm (LSBC). In addition, interactive correction background algorithm (IBC) was used to remove spectral interferences caused by Al lines on the analytical line of As. Limits of detection were 28.7 ng g-1 (As), 0.15 ng g-1 (Cd), 3.2 ng g-1 (Co), 7.3 ng g-1 (Pb), 40.1 ng g-1 (Se) and 24.5 ng g-1 (Sn). Micro-homogeneity studies were performed after using knife or cryogenic mills in order to establish minimal sample mass and homogeneity. Most accurate and precise results were obtained for sample masses above 0.2 mg for samples ground using knife mill with 30 mesh sieve or cryogenic mill. Accuracy and precision were evaluated by analyzing plant certified reference materials and results were in agreement with certified values at a 95% confidence level (t test). The proposed methods were applied for the determination of analytes in eighteen medicinal plant samples. The concentration ranged from 3.2 to 287.2 ng g-1 (Cd), 14.7 to 1195.7 ng g-1 (Co), 124.2 to 1946.1 ng g-1 (Pb) and 142.8 to 342.6 ng g-1 (Sn). Results were in agreement at a 95% confidence level... / Doutor

Sur près de 70 millions de personnes vivant avec l’épilepsie (PVE) dans le monde, 150 000 vivent au Pérou et 52 000 au Laos, avec un déficit de traitement (proportion de PVE qui ne reçoit pas de médicaments antiépileptiques adaptés), respectivement de 75% et 97%. Notre objectif principal était de caractériser l’utilisation des plantes médicinales dans la prise en charge de l’épilepsie. Les objectifs spécifiques visaient à : (i) faire l’état des lieux des connaissances sur les plantes médicinales utilisées comme antiépileptiques dans les pays d’Asie, d’Amérique Latine et d’Afrique, (ii) déterminer la place de la phytothérapie dans l’itinéraire thérapeutique des PVE (iii) évaluer l’efficacité pharmacologique d’extraits de plantes utilisées pour soigner l’épilepsie dans deux communautés rurales du nord du Pérou. D’abord, nous avons utilisé une approche bibliographique exhaustive de la littérature spécialisée. L’approche de terrain au Pérou et au Laos a combiné des outils épidémiologiques et ethnopharmacologiques. La troisième partie de ce travail était une étude pilote ayant pour objectif d’évaluer la faisabilité et la pertinence de réaliser des tests pharmacologiques de l’efficacité de plantes sur différents modèles d’épilepsie. Nous avons identifié 106 publications portant sur l’utilisation de plantes dans l’épilepsie(études réalisées entre 1982 et 2017). Sur le terrain, le pourcentage de recours aux remèdes à base de plantes était de 66,7% au Pérou et de 71,1% au Laos. Au total, 66 espèces végétales ont été citées par les patients et tradipraticiens au Pérou et 87 au Laos. Un effet antiépileptique potentiel avec les extraits de Valeriana officinalis et Melissa officinalis a été observé. La place de la médecine traditionnelle est telle dans l’itinéraire thérapeutique d’un patient épileptique qu’il serait intéressant qu’elle soit partie intégrante du système de soins. / Of the 70 million people living with epilepsy (PWE) worldwide, 150,000 live in Peru and 52,000in Laos, where the epilepsy treatment gap are respectively 75% and 97%.Our main objective was to characterize the use of medicinal plants in the management ofepilepsy. The specific objectives were: (i) to review the current state of knowledge onmedicinal plants used as antiepileptics in Asian, Latin American and African countries, (ii) to determine the place of phytotherapy in the therapeutic itinerary of PWE, (iii) to evaluate the pharmacological efficacy of plant extracts used to treat epilepsy in two rural communities in northern Peru.We first used a comprehensive bibliographic approach of the specialized literature. The field approach in Peru and Laos combined epidemiological and ethnopharmacological tools. The third part of this work was a pilot study to assess the feasibility and relevance of conducting pharmacological tests of plant efficacy in epilepsy models.We identified 106 publications on the use of plants in epilepsy between 1982 and 2017. Thepercentage of herbal remedies used was 66.7% in Peru and 71.1% in Laos. A total of 66 plant species were mentioned by patients and traditional healers in Peru and 87 in Laos. A potential antiepileptic effect with extracts of Valeriana officinalis and Melissa officinalis has been observed. The place of traditional medicine is such that it would be interesting if it were an integral part of the health care system.

Systematische Übersichtsarbeiten klinischer Studien spielen bei der Beurteilung der Wirksamkeit prophylaktischer und therapeutischer Interventionen eine zentrale Rolle. Übersichtsarbeiten (im folgenden synonym mit der Bezeichnung Review gebraucht) werden dann als systematisch bezeichnet, wenn sie vordefinierte und transparente Methoden bzgl. Literatursuche, Studienselektion und Studienbewertung verwenden. Systematische Übersichtsarbeiten, bei denen die einbezogenen Studien (Primärstudien) in einer integrierenden statistischen Analyse wie eine einzige große Studie ausgewertet (gepoolt) werden, bezeichnet man als Meta-Analysen. Die vorliegende Monographie präsentiert 1. zwei größere eigene systematische Übersichtsarbeiten als Anwendungsbeispiele (Kapitel 2), 2. drei empirisch-methodische Untersuchungen zum Thema Studienqualität und deren Auswirkung auf das Ergebnis (Kapitel 3) und 3. eine Analyse der vorliegenden systematischen Übersichtsarbeiten in den Bereichen Akupunktur, Phytotherapie und Homöopathie (Kapitel 4). Sie versteht sich als Beitrag zur methodischen Diskussion und Entwicklung von systematischen Übersichtsarbeiten. Im ersten Anwendungsbeispiel wurde untersucht, ob Propranolol, Metoprolol und Flunarizin bei der prophylaktischen Behandlung der Migräne wirksamer sind als Placebo, ob die einzelnen Substanzen ähnlich wirksam sind bzw. wie wirksam sie im Vergleich zu anderen Medikamenten sind. Eingeschlossen wurden randomisierte klinische Studien an erwachsenen Migränepatienten mit mindestens 4 Wochen Dauer, in denen eine oder mehrere der genannten Substanzen mit Placebo oder einem anderen Medikament verglichen wurden. Die Literatursuche erfolgte in Medline und der Cochrane Library sowie über das Durchsuchen von Literaturverzeichnissen. Informationen zu Patienten, Interventionen und Ergebnissen wurden mit Hilfe eines vorgetesteten Formulars extrahiert. Die Qualität der Studien wurde mit drei Skalen beurteilt. Soweit möglich erfolgten Effektstärkeberechnungen für Therapieerfolg, Attackenhäufigkeit und Zahl der Patienten mit Nebenwirkungen. Eine quantitative Meta-Analyse wurde aufgrund der inkonsistenten und häufig inadäquaten Ergebnispräsentation in den Primärstudien nicht durchgeführt. Insgesamt entsprachen 57 Studien zu Propranolol (mit 25 Vergleichen vs. Placebo und 46 Vergleichen mit anderen Medikamenten), 16 zu Metoprolol (mit 4 Vergleichen vs. Placebo und 17 vs. andere) und 40 zu Flunarizin (mit 10 Vergleichen vs. Placebo und 36 vs. andere) den Einschlusskriterien. Die Qualität der Studien war in vielen Fällen unbefriedigend. Hauptproblem war die unzureichende Beschreibung von Studienabbrüchen und -ausschlüssen bzw. deren Berücksichtigung in der Analyse. Alle drei Substanzen waren in Bezug auf Therapieerfolg und Attackenzahl Placebo überlegen. Langzeiteffekte über 6 Monate hinaus und nach Absetzen können nicht sicher beurteilt werden. Mit wenigen Ausnahmen ergaben sich in Vergleichen mit anderen Substanzen keine signifikanten Unterschiede; die meisten Studien hatten jedoch zu geringe Fallzahlen, um potenziell relevante Unterschiede ausschließen zu können. Insgesamt kann die Wirksamkeit von Propranolol, Metoprolol und Flunarizin in der Migräneprophylaxe aufgrund der Konsistenz der Ergebnisse trotz der häufig unbefriedigenden Studienqualität als belegt angesehen werden. Im zweiten Anwendungsbeispiel wurde untersucht, ob Hypericumextrakte (Hypericum perforatum, Johanniskraut) bei der Behandlung von Depressionen (a) wirksamer sind als Placebo, (b) ähnlich wirksam sind wie synthetische Antidepressiva und (c) nebenwirkungsärmer sind als synthetische Antidepressiva. Eingeschlossen wurden randomisierte Studien an depressiven Patienten mit einer Therapiedauer von mindestens 4 Wochen, in denen ein Hypericummonoextrakt mit Placebo oder einem anderen Antidepressivum verglichen wurde. Die Literatursuche erfolgte in verschiedenen Datenbanken, über das Prüfen von Literaturverzeichnissen und über Kontakte mit Experten und Herstellern. Hauptzielkriterium für die Beurteilung der Wirksamkeit war der Responderanteil (meist definiert als ein Rückgang um 50% oder auf einen Wert von max. 10 auf der Hamilton Rating Scale for Depression). 33 Studien (23 Vergleiche mit Placebo, 12 mit anderen Antidepressiva) entsprachen den Einschlusskriterien. Im Vergleich zu Placebo erwiesen sich die Hypericumextrakte als deutlich überlegen, allerdings war der Effekt in neueren Studien und in Studien mit schwerer depressiven Patienten geringer ausgeprägt. Im Vergleich zu älteren und neueren Antidepressiva waren Hypericumextrakte ähnlich effektiv; Nebenwirkungen waren jedoch signifikant seltener. Aufgrund der vorliegenden Daten kann die Wirksamkeit von Hypericumextrakten bei leichten bis mittleren Depressionen als nachgewiesen gelten. Die Heterogenität in den Ergebnissen der einzelnen Studien deutet jedoch darauf hin, dass verschiedene Faktoren (möglicherweise Extraktwahl, Dosierung, Patientenklientel) die Effektivität beeinflussen. Die erste der drei empirisch-methodischen Analysen in Kapitel 3 untersucht die methodische Qualität randomisierter Studien zu Akupunktur, Phytotherapie und Homöopathie. Insgesamt 207 randomisierte Studien wurden einbezogen; diese waren im Rahmen von 5 systematischen Übersichtsarbeiten gesammelt und bewertet worden. Die Beurteilung der methodischen Qualität erfolgte mithilfe einer validierten Skala (Jadadscore) und Einzelitems. Die methodische Qualität der bewerteten Studien war in hohem Maße variabel, die Mehrzahl der Studien hatte relevante Schwächen. Hauptprobleme waren die Beschreibung der Verblindung der Gruppenzuteilung sowie von Studienabbrüchen und -ausschlüssen. Phytotherapiestudien hatten im Mittel bessere Qualität als Homöopathie- und Akupunkturstudien. Publikationen in einer Medline-gelisteten Zeitschrift, in jüngerer Vergangenheit und in englischer Sprache hatten im Mittel bessere Qualität als ältere, nicht in gelisteten Zeitschriften und in anderen Sprachen veröffentlichte Arbeiten. Die Qualitätsscores der komplementärmedizinischen Studien waren im Schnitt ähnlich wie diejenigen von Studien zu konventionellen Interventionen, die in ähnlichen Analysen bewertet worden waren. In der zweiten empirisch-methodischen Untersuchung wurde geprüft, ob ein Zusammenhang zwischen methodischer Qualität und Studienergebnis in 89 placebokontrollierten Studien zur Homöopathie besteht. Die Beurteilung der methodischen Qualität erfolgte mit zwei Skalen und Einzelkriterien. Ein potenzieller Zusammenhang zwischen methodischer Qualität wurde mit drei Methoden untersucht: Prüfung eines Zusammenhangs zwischen einzelnen Qualitätskomponenten und Studienergebnis (Komponentenanalyse), Vergleich von Studien, die eine bestimmte Minimumpunktzahl auf den Qualitätsskalen erreichten bzw. nicht erreichten (Mindestscoreanalyse), und konsekutiver Einschluss von Studien in die Meta-Analyse entsprechend den erreichten Qualitätsscores (kumulative Meta-Analyse). Mit allen drei Methoden ergab sich, dass bessere Studien weniger optimistische Ergebnisse erbracht hatten. In der dritten empirisch-methodischen Untersuchung wurde am Beispiel der klinischen Studien zu Akupunktur bei Kopfschmerzen die zuletzt angesprochene Thematik auch auf nichtkontrollierte Studien ausgeweitet. Es wurde geprüft, inwieweit sich randomisierte und nichtrandomisierte Studien in Bezug auf Patienten, Interventionen, design-unabhängige Qualitätsaspekte und Ergebnisse unterscheiden. 59 Studien (24 randomisierte und 35 nichtrandomisierte) konnten in die Analyse einbezogen werden. Randomisierte und nichtrandomisierte Studien unterschieden sich bzgl. Patienten und Interventionen zum Teil deutlich. Nichtrandomisierte Studien hatten im Schnitt schlechtere Qualität, jedoch gab es unter den randomisierten Studien sehr schlechte und unter den nichtrandomisierten auch gute Studien. Ein Randomisationsdesign und bessere Qualität waren mit weniger positiven Ergebnissen assoziiert. Die nichtrandomisierten Studien nutzten potenzielle Vorteile (bzgl. Fallzahl, Langzeitbeobachtung, Repräsentativität und Untersuchung prognostischer Faktoren) kaum. Der Einbezug nichtrandomisierter Studien in einen systematischen Review zur Evidenz für die Wirksamkeit der Akupunktur bei Kopfschmerzen hätte daher die Schlussfolgerungen kaum beeinflusst. In Kapitel 4 wurden die bis Frühjahr 2001 verfügbaren systematischen Übersichtsarbeiten zu Akupunktur, Phytotherapie und Homöopathie zusammengestellt. Die Suche erfolgte primär über die Datenbank des Bereichs Komplementärmedizin der Cochrane Collaboration und in Medline. Um als systematisch klassifiziert zu werden, mußte ein Review mindestens eines der folgenden drei Kriterien erfüllen: explizite Beschreibung von Literatursuche und Einschlusskriterien und/oder formale Beurteilung der methodischen Qualität der eingeschlossenen Studien und/oder Beschreibung und Durchführung einer quantitativen Meta-Analyse. 39 Reviews zur Akupunktur, 58 zur Phytotherapie und 18 zur Homöopathie entsprachen den Einschlusskriterien. Positive Schlussfolgerungen waren im Bereich Phytotherapie am häufigsten, bei der Akupunktur am seltensten. Die Qualität der Reviews war variabel, viele hatten jedoch relevante methodische Schwächen. Siebzehn Fragestellungen wurden von mehr als einer systematischen Übersicht bearbeitet. Die Zahl der eingeschlossenen Primärstudien variierte bei 10 von 17 Vergleichen um mehr als 50%. Unterschiede in den Einschlusskriterien und der Literatursuche erschienen als wahrscheinlichste Ursache für die Diskrepanzen, Unterschiede in der methodischen Qualität der Reviews schienen dagegen eine geringe Rolle zu spielen. Die Schlussfolgerungen waren trotz der Diskrepanzen häufig ähnlich. Die vorgelegten Anwendungsbeispiele, empirisch-methodischen Untersuchungen und die Zusammenstellung zu Akupunktur, Phytotherapie und Homöopathie demonstrieren Stärken und Schwächen von systematischen Reviews. Trotz der zu erwartenden methodischen Weiterentwicklungen werden derartige Übersichtsarbeiten auch in Zukunft mit Sorgfalt und Zurückhaltung interpretiert werden müssen. / Systematic reviews of randomized controlled trials have a central role in the assessment of the effectiveness of health care interventions. Reviews are considered systematic if they describe and use adequate methods for literature search, study selection, quality assessment and summary of study results. Meta-analyses are systematic reviews if the results of the included primary studies are pooled to calculate an overall effect size measure. This monograph reports 1) two major own systematic reviews as examples (chapter 2); 2) three empirical studies on the issue of study quality and its effect on outcome (chapter 3); and 3) an analysis of the available systematic reviews on acupuncture, herbal medicines, and homeopathy (chapter 4). The reported work aims to contribute to the further development of the methodology of systematic reviews. The first systematic review in chapter 2 investigates whether there is evidence that the prophylactic treatment of migraine with propranolol, metoprolol and flunarizine is more effective than placebo and how these drugs compare to each other and to other drugs for migraine prophylaxis. To be included studies had to be randomized trials of at least 4 weeks duration in adult migraine patients comparing one of the three drugs with placebo or another drug treatment. Studies were identified through the database Medline, the Cochrane Library, and screening of bibliographies. Information on patients, interventions, methods and results was extracted in a standardized manner. Methodological quality was assessed with three checklists. As far as possible effect size estimates were calculated for single trials for the outcomes response, frequency of attacks, and number of patients with side effects. A pooled effect size estimate was not calculated due to the inconsistent and often insufficient presentation of results in the primary studies. A total of 57 trials on propranolol (with 25 comparisons vs. placebo and 46 comparisons vs. other drugs), 16 on metoprolol (4 vs. placebo and 17 vs. other drugs), and 40 on flunarizine (10 vs. placebo and 36 vs. other drugs) met the inclusion criteria. The majority of trials had relevant methodological shortcomings. The major problems were the description of drop-outs and withdrawals and the lack of intention to treat analyses. All three drugs have been shown to be superior to placebo in respect to response rates and frequency of attacks. There is insuccifient data to assess long-term effects and duration of the treatment effects after stopping prophylaxis. Apart from few exceptions there were no statistically significant differences in effectiveness in comparisons with other drugs. However, most trials had insufficient power to rule out potentially relevant differences. Overall, despite considerable methodological shortcomings the short-term effectiveness of propranolol, metoprolol and flunarizine for migraine prophylaxis has been proven beyond reasonable doubt. The second systematic review in chapter 2 investigates whether extracts from St. John s wort (Hypericum perforatum L.) are a) more effective than placebo, b) similarly effective as synthetic drugs, and c) have less side effects than synthetic drugs in the treatment of patients suffering from depression. Randomized trials in depressed patients were included if treatment with a hypericum mono-preparation for at least 4 weeks was compared with placebo or a synthetic antidepressant. Trials were identified from a variety of databases, by screening bibliographies, and through contacts with manufacturers and experts in the field. Main outcomes measure to assess effectiveness was the proportion of responders (mostly defined as patients with a 50% reduction vs. baseline or less than 10 points on the Hamilton Rating Scale for Depression). 33 trials (with 23 comparisons with placebo, and 12 with synthetic antidepressants) met the inclusion criteria. Hypericum extracts were markedly superior to placebo; however, the effect was smaller in more recent trials and in trials with patients suffering from more severe depression. The available trials indicate that hypericum extracts are similarly effective as both older and newer synthetic antidepressants in patients with mild and moderate depression and have less side effects. The heterogeneity of the observed effects suggests that a number of variables (mainly patient characteristics and quality of trials) influences outcomes. The first of the three empirical studies on review methodology in chapter 3 investigates the quality of randomized trials of acupuncture, herbal medicines, and homeopathy. A total of 207 studies originally collected for 5 systematic reviews were included in the analyses. Methodological quality was assessed with the validated scale by Jadad et al. and a number of single quality criteria. The quality of the included trials was highly variable but the majority of trials had relevant shortcomings. Major problems were the description of concealment of randomisation, and of drop-outs and withdrawals. In average, trials on herbal remdies had better quality than acupuncture and homeopathy trials. Publication in a Medline-listed journal, in more recent years and in English language was associated with better quality ratings. Compared to a sample of studies form conventional medicine from a similar analysis the average quality of trials in the three complementary medicine areas was very similar. The second study on methodology investigates whether there is an association between methodological quality and results in 89 placebo-controlled trials on homeopathy. Methodological quality was assessed with two score systems and single quality criteria. A potential association of quality and outcome was investigated with three approaches: 1) Testing whether the fulfillment of single criteria had impact on the pooled effect estimate (component analysis); 2) whether results of trials scoring a minimal number of points were different from those of trials with lower scores (minimal score analysis); 3) entering studies consecutively into the meta-analysis depending on the number of score points achieved (cumulative meta-analysis). With all three methods trials with better methodological quality were shown to yield less optimistic results. In the third study on methodology it was investigated whether randomized and non-randomized trials of acupuncture for chronic headache differ in regard to patients, interventions, design-independent quality aspects and results. 59 studies (24 randomized and 35 non-randomized) met the inclusion criteria. Randomized and non-randomized differed significantly regarding patient characteristics and interventions. In average, non-randomized studies had lower quality; however, there were also good quality non-randomized studies and randomized trials with very low quality. Studies with random assignment and better quality in general yielded less optimistic results. The non-randomized studies rarely took advantage of the possible strengths of such designs (in respect to sample size, long-term observation, representative sampling and investigation of prognostic factors). In the case of acupuncture for chronic headache the inclusion of non-randomized studies would not have altered the conclusions of a systematic review on effectiveness. In chapter 4 the systematic reviews on acupuncture, herbal medicines and homeopathy available until spring 2001 have been summarized. Eligible reviews were identified mainly through searches in the database of the Cochrane Collaboration s Complementary Medicine Field and in Medline. To be included a review had to meet at least one of the following criteria: explicit description of literature search and inclusion criteria; formal assessment of the methodological quality of the included primary studies; performance of a quantitative meta-analysis. 39 reviews on acupuncture, 58 on herbal medicines and 18 on homeopathy met the inclusion criteria. Positive conclusions were most frequent in herbal medicine, and rare in acupuncture. The quality of reviews was variable, but many had relevant shortcomings. 17 topics were adressed by more than one systematic reviews. In 10 of these 17 topics the number of included primary studies varied by more than 50%. Differencs in selection criteria and the literature search were the most likely cause for discrepancies. Differences in methodological quality had little impact. Conclusions were often similar despite discrepanices in methods and results. The examples and empirical studies on methodology presented in this monograph demonstrate the strengths and limitations of systematic reviews. Although it can be expected that the methods of systematic reviews will be improved in the future this kind of research will still have to be interpreted with great caution.

Nesta tese, objetivou-se avaliar a ação antibacteriana in vitro dos extratos das plantas medicinais mais utilizadas no município de Foz do Iguaçu e região,sobre Pseudomonas aeruginosa, Klebsiella pneumoniae e Staphylococcus aureus. As plantas mais utilizadas no município de Foz do Iguaçu e região, que são: Mikania glomerata (Guaco), Mentha piperita (Hortelã), Maytenus officinalis (Espinheira-santa) e Aloe vera (Babosa) foram preparadas como fitoterápicos e cedidas pelo Horto de plantas medicinais da hidroelétrica Itaipu Binacional. Para esta avaliação foram preparados extratos hidroalcóolico a 70% por percolação e extratos aquosos por infusão destas plantas. Os extratos hidroalcóolicos foram levados a rota-evaporação e os aquosos ao liofilizador. Também foi extraído o óleo essencial da hortelã pimenta empregando o método de destilação por arraste a vapor d\'água em aparelho de Clevenger seguido da cromatografia por camada delgada. A atividade antibacteriana dos extratos foi testada sobre as linhagens padrão de Pseudomonas aeruginosaATCC (27853), Staphylococcus aureusATCC (25923) e Klebsiella pneumoniaeATCC (13883). A determinação das concentrações inibitórias mínimas (CIM)e das concentrações bactericidas mínimas (CBM)dos extratos vegetais sobre as linhagens bacterianas foram determinadas pelo método de microdiluição em microplaca segundo a metodologia preconizada pelo Clinical and Laboratory Standards Institute (CLSI). Entre os extratos hidroalcoólicos e aquosos das plantas testados, somente o extrato hidroalcoólico de Guaco (Mikania glomerata) apresentou atividade antibacteriana positiva sobre Staphylococcus aureus ATCC 25923, na concentração de 200 µg. Também foi observada atividade antibacteriana do óleo essencial da Hortelã pimenta (Mentha piperita) sobre Staphylococcus aureus ATCC 25923, com CIM de 1000 µg/mL, Klebsiella pneumoniaeATCC 13883 com CIM de 2000 µg/mL e CBM > 2000 µg/mL e Pseudomonas aeruginosa ATCC 27853 com CIM e CBM de 2000 µg/mL.Os fitoterápicos mais prescritos nas unidades básicas de saúde do município de Foz do Iguaçu são o Guaco e a Espinheira Santa. Considerando os resultados dos esperimentos que apontam a atividade antibacteriana dos extratos das plantas medicinais mais utilizadas em Foz do Iguaçu e região, pode-se indicar que os estratos de Guaco, Hortelã-pimenta, Espinheira Santa e Babosa, bem como óleo essencial de hortelã-pimeta prometem uma altermativa possível no tratamento de doenças infecciosas / In this thesis, the objective was to evaluate the in vitro antibacterial action of extracts of medicinal plants most used in the municipality of Foz do Iguaçu and region, on Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus. The most used plants in the municipality of Foz do Iguaçu and region, which are: Mikania glomerata (Guaco), Mentha piperita (Hortelã pimenta), Maytenus officinalis (Espinheira-santa) and Aloe vera (Babosa) were prepared as phytotherapics and yielded by Horto de plants of the Itaipu Binacional hydroelectric plant. For this evaluation 70% hydroalcoholic extracts were prepared by percolation and aqueous extracts by infusion of these plants. The hydroalcoholic extracts were rotoevaporated and the aqueous extracts were added to the lyophilizer. The peppermint essential oil was also extracted using the water vapor distillation method in a Clevenger apparatus followed by thin layer chromatography. The antibacterial activity of the extracts was tested on the standard strains of Pseudomonas aeruginosa ATCC (27853), Staphylococcus aureus ATCC (25923) and Klebsiella pneumoniae ATCC (13883). The determination of minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (CBM) of the plant extracts on the bacterial strains were determined by the microplate microdilution method according to the methodology recommended by the Clinical and Laboratory Standards Institute (CLSI). Among the hydroalcoholic and aqueous extracts of the plants tested, only the hydroalcoholic extract of Guaco (Mikania glomerata) showed positive antibacterial activity on Staphylococcus aureus ATCC 25923, in the concentrations of 400 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg and 200 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg. The other hydroalcoholic and aqueous extracts presented bacteriostatic activity. Antimicrobial activity of peppermint (Mentha piperita) essential oil on Staphylococcus aureus ATCC 25923, with MIC of 1000 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg / mL, Klebsiella pneumoniae ATCC 13883 with MIC of 2000 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg / mL and CBM >= 2000 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg / mL and Pseudomonas aeruginosa ATCC 27853 with MIC and CBM of 2000 ?g and 200 ?g. The other hydroalcoholic and aqueous extracts presented bacteriostaticg / ml. The most prescribed herbal medicines in the basic health units of the municipality of Foz do Iguaçu are Guaco and Espinheira Santa. Considering the results of the experiments that point out the antibacterial and bacteriostatic activity of the most used medicinal plants in Foz do Iguaçu, it is possible to indicate that the extracts of Guaco, Peppermint, Espinheira Santa and Babosa as well as peppermint essential oil can serve as an alternative in the treatment of infectious diseases

Submitted by PPG Odontologia (odontologia-pg@pucrs.br) on 2018-06-27T15:29:59Z No. of bitstreams: 1 JULIANE_DE_QUADROS_DE_BORTOLLI_DIS.pdf: 1560215 bytes, checksum: 631d814b124322abfc0f1aaf6c260aa3 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-07-04T13:31:02Z (GMT) No. of bitstreams: 1 JULIANE_DE_QUADROS_DE_BORTOLLI_DIS.pdf: 1560215 bytes, checksum: 631d814b124322abfc0f1aaf6c260aa3 (MD5) / Made available in DSpace on 2018-07-04T13:41:52Z (GMT). No. of bitstreams: 1 JULIANE_DE_QUADROS_DE_BORTOLLI_DIS.pdf: 1560215 bytes, checksum: 631d814b124322abfc0f1aaf6c260aa3 (MD5) Previous issue date: 2018-03-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The oral mucosa is a frequent site for traumatic ulcers, which can be caused by mechanical or chemical agents. There is no a gold standard treatment for the management of these lesions, which can be painful and have a negative impact on quality of life. Although studies suggest the use of chamomile and curcumin in the treatment of ulcerated lesions of the oral mucosa, there is no a consensus as to its efficacy in the management of these lesions. The literature has shown that both phytotherapics modulate the levels of inflammatory cytokines such as TNF-? and IL-1?. Therefore, in the first article of this dissertation, an integrative literature review was conducted investigating the efficacy of topical interventions with chamomile and curcumin in the management of pain and in the healing of ulcerated lesions of the oral mucosa. Few controlled clinical trials have been found and they suggest a beneficial effect of chamomile and curcumin in the healing process and in the reduction of symptoms in patients with oral mucositis, recurrent aphthous ulceration and oral lichen planus. Studies in animal demonstrated a reduction in the inflammatory wound healing phase, increased deposition of collagen fibers and epithelization. The second article of this dissertation presents an experimental study, which was carried out to evaluate the effect of the 10% chamomile extract and 2% curcumin in the repair of traumatic ulcers on tongue of rats. Thirty-nine Wistar rats were divided into three groups: control, chamomile extract and curcumin. A 5-mm-diameter excisional wound was made in the center of ventral tongue. The substances were applied as a gel twice daily for five days. The animals were euthanized 7 days after the lesions were made. In the macroscopic evaluation, there was no significant difference among chamomile, curcumin and control groups regarding to area of residual ulcer (P= 0.20). Histological analysis also showed no difference among the groups regarding newly formed epithelium area (P=0.97) or intensity of inflammatory infiltrate (P=0.72). Considering the experimental model, the findings of the present study showed that 10% chamomile extract and 2% curcumin did not accelerate the repair of traumatic oral ulcers or reduced the intensity of the inflammatory infiltrate, contradicting previous findings in the literature. / A mucosa bucal ? um s?tio frequente para a ocorr?ncia de ?lceras traum?ticas, as quais podem ser causadas por agentes mec?nicos ou qu?micos. At? o momento, n?o existe um tratamento padr?o-ouro para o manejo dessas les?es, as quais podem ser dolorosas e ter impacto negativo na qualidade de vida dos pacientes. Apesar de estudos sugerirem a utiliza??o da camomila e da curcumina no tratamento de les?es ulceradas da mucosa bucal, n?o h? consenso sobre a sua efic?cia no manejo dessas les?es. A literatura tem demonstrado que ambos os fitoter?picos modulam os n?veis de citocinas pr?-inflamat?rias como TNF-? e IL-1?. No primeiro artigo desta disserta??o foi realizada uma revis?o integrativa da literatura investigando a efic?cia das interven??es t?picas com camomila e curcumina no manejo da dor e na cicatriza??o de les?es ulceradas da mucosa bucal. Poucos ensaios cl?nicos controlados foram encontrados e estes sugerem um efeito ben?fico da camomila e da curcumina no processo cicatricial e na redu??o da sintomatologia em pacientes com mucosite oral, ulcera??o aftosa recorrente e l?quen plano oral. Estudos em modelos animais investigando os efeitos desses fitoter?picos em ?lceras traum?ticas bucais demonstraram redu??o da fase inflamat?ria do processo de cicatriza??o, maior deposi??o de fibras col?genas e epiteliza??o. O segundo artigo desta disserta??o apresenta um estudo experimental, que foi realizado com o objetivo de avaliar o efeito do extrato de camomila 10% e curcumina 2% no reparo de ?lceras traum?ticas em l?ngua de ratos. Trinta e nove ratos Wistar foram distribu?dos em tr?s grupos: controle, extrato de camomila e curcumina. Uma ?lcera de 5 mm de di?metro foi confeccionada no ventre lingual de cada animal. As subst?ncias foram aplicadas na forma de gel, duas vezes ao dia, durante cinco dias. Os animais foram eutanasiados sete dias ap?s a confec??o das les?es. Na avalia??o macrosc?pica, n?o houve diferen?a entre os grupos camomila, curcumina e controle quanto ? ?rea de ?lcera remanescente (P=0.20). A an?lise histol?gica tamb?m n?o demonstrou diferen?a entre os grupos quanto ? ?rea de tecido epitelial neoformado (P=0.97) ou intensidade do infiltrado inflamat?rio (P=0.72). Considerando o modelo experimental empregado, os achados do presente estudo demonstraram que extrato de camomila 10% e a curcumina 2% n?o aceleraram o reparo de ?lceras mecanicamente induzidas na mucosa bucal nem reduziram a intensidade do infiltrado inflamat?rio, contrariando achados pr?vios na literatura.

A mastite é o processo inflamatório da glândula mamária que constitui uma doença comum no setor pecuário leiteiro, evidenciado principalmente durante o período de lactação de vacas leiteiras e, cujos principais agentes etiológicos são o Staphylococcus aureus, Streptococcus sp. e espécies de coliformes. Os protocolos de combate à mastite preconizam o uso de antibióticos, o que pode trazer riscos à saúde dos animais e humana. Dependendo do agente patogênico, esta estratégia é considerada frequentemente ineficiente e de alto custo, o que leva pesquisadores a buscarem alternativas à antibioticoterapia, testando princípios ativos homeopáticos, fitoterápicos e micro-organismos probióticos, que apresentam potencial como alternativa promissora para a terapia quimioterápica. As formulações contendo bactérias probióticas podem representar uma alternativa de baixo custo para o combate à mastite, com a vantagem de não estimular a resistência entre os patógenos. Desta forma, este estudo baseou-se na avaliação in vitro do potencial antimicrobiano de cepas de Lactobacillus frente a agentes patógenos causadores de mastite, bem como na avaliação in vivo da inocuidade de formulações contendo probióticos em vacas em lactação. Para tanto, 6 cepas de Lactobacillus, na forma isolada e na forma de pool foram avaliadas quanto à capacidade inibitória sobre Streptococcus agalactiae (cepas 1 e 2) por diferentes técnicas de difusão em ágar e co-cultura em meio líquido. A inibição pela técnica de co-cultura foi avaliada mediante determinação de células viáveis das cepas patogênicas por reação em cadeia da polimerase quantitativa em tempo real (RT-qPCR). De acordo com os resultados observados no estudo de inibição in vitro, foram preparadas formulações probióticas utilizando dois veículos diferentes, sendo uma emulsão (Formulação A) e outra utilizando um veículo de um medicamento comercial (Formulação B) adicionadas de células de Lactobacillus na forma isolada e na forma de pool. Estas formulações foram avaliadas quanto à inocuidade em 17 vacas separadas em grupos sob regime de 1 ordenha/dia (A e B1) e 2 ordenhas/dia (B2), mediante a aplicação das formulações A e B contendo 109 ou 106 células/dose. Ao longo do cada tratamento, foram avaliados parâmetros fisiológicos como temperatura do úbere, presença de hiperemia, inchaço e manifestação de dor, contagem de células somáticas (CCS), California Mastitis Test (CMT), bem como condutividade elétrica e aspecto do leite e avaliação microbiológica. O estudo de inibição em co-cultura mostrou que as cepas de Lactobacillus avaliadas inibiram o crescimento das cepas patogênicas, em níveis superiores a 75% após 72 horas de ensaio. Após 24 horas, os maiores níveis de inibição sobre S. agalactiae cepa 1 foram exercidos pelas cepas L. acidophilus ATCC 4356, L. fermentum ATCC 9338, com índices de inibição de 99,79% e 99,91%, respectivamente. Nas mesmas condições a inibição de S. agalactiae cepa 2 por L. acidophilus ATCC 4356 e L. delbrueckii UFV H2B20 foi de 86,7% e 79,5%, respectivamente. Estes resultados indicam o potencial de inibição das cepas de Lactobacillus estudadas sobre S. agalactiae causadora de mastite. Estudos de inocuidade demonstraram que as formulações probióticas (A e B) induziram reação inflamatória no úbere dos animais, confirmado pelo aumento significativo da condutividade elétrica do leite (CEL), CCS (p < 0,05), bem como do escore de CMT. As formulações testadas no presente estudo não se mostraram adequadas ao emprego terapêutico na cura da mastite, uma vez que no teste de inocuidade provocaram processos inflamatórios nas glândulas mamárias dos animais testados. / Bovine mastitis is a common disease in dairy farms, usually observed during lactation period in dairy cattle, whose main etiological agents are Staphylococcus aureus, Streptococcus sp. and coliform species. Current protocols of mastitis therapy include the use of antibiotics, which may represent serious risks for both animal and human health. Depending on the pathogen specie, this strategy is often considered ineffective and highly expensive, what stimulates scientists to search for alternative treatments, including homeopathic or phytotherapic drugs or probiotic microorganisms, which may be a good alternative to chemotherapeutic drugs. Formulations containing probiotic bacteria may represent a low-cost alternative for mastitis treatment. The present study aimed to assess in vitro ability of Lactobacillus strains to inhibit mastitis pathogens growth, as well as in vivo assessment of the safety of formulations containing probiotics in cows with predisposition or affected by mastitis. Thus, six Lactobacillus strains were evaluated on their inhibitory ability over Streptococcus agalactiae (strains 1 and 2) by techniques of agar diffusion and co-culture in liquid medium. The effectiveness of the co-culture was evaluated by enumerating viable pathogen cells through quantitative polymerase chain reaction (qPCR). According to the results of in vitro study, two formulations were prepared based on a probiotic emulsion (formula A) and a commercial matrix (formula B) containing Lactobacillus cells (isolated or pool). Safety tests for the formulations were performed in 17 cows, which were separated into two groups under 1 milking/day (A and B1) and another group submitted to 2 milkings/day (B2), through the application of preparation A or preparation B containing 109 or 106 cells/dose. In each treatment, every cow was assessed on physiological parameters including temperature, presence of hyperemia, swelling and pain, somatic cell count (SCC), electrical conductivity and aspect of milk (color and clots/lumps formation) and microbiological evaluation. The results of the inhibition studies in co-culture showed that all Lactobacillus strains inhibited the growth of pathogenic strains, in at least 75% after 72 h of test. The highest levels of inhibition of S. agalactiae strain 1 were performed by strains L. acidophilus ATCC 4356 and L. fermentum ATCC 9338, at level of 99.79% and 96.70% after 24h, respectively. For the same period, the inhibition of S. agalactiae strain 2 by L. acidophilus ATCC 4356 and L delbrueckii UFV H2B20 was 86.7 % and 79.5%, respectively. These results showed the potential of Lactobacillus strains studied concerning to the inhibition of mastitis caused by Streptococcus agalactiae. Safety studies showed that both formulations evaluated induced inflammatory response in the udder of the animals, what was confirmed by significant raise on electric conductivity and SCC (p < 0,05) and CMT scores on milk. All formulations were considered inappropriate to be used in mastitis therapeutics, once they failed at safety tests, inducing mastitis on tested animals.

O homem sempre buscou nas plantas uma alternativa no tratamento para resolver suas patologias. As plantas medicinaisvêm sendo utilizados para fins terapêuticos e em alguns casos sem conhecer o princípio ativo e a toxicidade destes, o Anacardium occidentale L (caju) planta com pseudofruto suculento, é característico da região tropical do Brasil. O Anacardium microcarpum D (cajui) presente na região dos cerrados da Amazônia e das regiões Nordeste, Centro-Oeste e Sudeste brasileiro. O metabolismo secundário das folhas, cascas e pseudofruto dessas plantas produz compostos fenólicos,com atividades anti-inflamatória, antioxidante e antimicrobiana. Os objetivos deste estudo foi realizar a prospecção fitoquímica,avaliar a atividade antimicrobiana, antioxidante e anti-inflamatória, in vivo e in vitrode extratos das folhas do Anacardium occidentaleL (caju)e Anacardium microcarpumD (cajui). Os resultados estão divididos em três artigos, sendo composto poruma revisão sistemática das ações antimicrobiana e antioxidantedo caju (Anacardium occidentale), do cajuí (Anacardium microcarpum)e dopequí (Caryocar brasiliense), edois artigos originais, um com ensaios antioxidante e antimicrobiano in vitroe outro com ensaios do controle metabólico, estresse oxidativo e hepatoxicidadein vivo. Artigo 1. Antioxidant and antimicrobial activities of crude extracts and fractions of cashew (Anacardium occidentale L.), cajui (Anacardium microcarpum) and pequi (Caryocar brasiliense C). A systematic review. Elaborou-se uma revisão sistemática dos trabalhos que apresentassem resultados de ensaios com extratos das partes das espécies vegetais acima mencionadas. Foram analisadas as bases de dados Pubmed/Medline, Biblioteca virtual em Saúde (Lilacs e Scielo) e Science Direct.De um total de 425 artigos, 32 foram incluídos, sendo19 estudos com Anacardiumoccidentale,3 com Anacardiummicrocarpum e 10 comCaryocar brasiliense.A técnica para avaliação da capacidade antioxidante in vitro mais utilizada foi de DPPH e da atividade microbiológica foi a concentração inibitória mínima e difusão em discos. A maioria dos ensaios que foram descritos demonstraram resultados potencialmente positivos às ações antioxidantes e antimicrobianas quando da utilização das espécies estudadas. Os resultados trazem importantes dados e perspectivas para a utilização de produtos naturais que possam contribuir para o tratamento de várias enfermidades, destacando-se principalmente as doenças e agravos não transmissíveis. Artigo 2. Caracterização fitoquímica e análise das atividades antimicrobiana e antioxidante de folhas de Anacardium occidentale L. e Anacardium microcarpum D., in vitro. A partir das folhas secas trituradas foram preparados extratos brutos etanólicos de A. occidentale L e A. microcarpum D. Realizou-se a composição centesimal da matéria seca, para determinar a quantidade de lipídios, proteínas, cinzas, umidade e carboidratos totais. A cromatografia de camada delgada foi a técnica de escolha para a prospecção fitoquímica. Por espectrofotometria e regressão linear foram quantificados os fenólicos e flavonoides totais dos extratos. A Cromatografia Líquida de Alta Permance (HPLC) foi utilizada para verificar a presença de ácido anacárdico. No ensaio antioxidante utilizaram-se as metodologias de captura de radicais estáveis DPPH (2,2-difenil-1-picrilidrazila)e a atividade antioxidante por meio da captura do radical 2,2´- azino-bis (3-etilbenzotiazolina-6-ácido sulfônico)-ABTS. Para os testes microbiológicos utilizou-se a concentração inibitória mínima em microplacas de 96 poços a partir de cepas multirresistentes de origem hospitalar.Os extratos etanólicos de Anacardium occidentaleL e AnacardiummicrocarpumDapresentaram captura de radicais DPPH e ABTS semelhantes e foram encontrados 2,32 e 1,64 mg EAG (equivalentes de ácido gálico) de fenólicos totais e valores de flavonóides totais de 0,34 e 0,29 mg ER (equivalentes de rutina), respectivamente.Na prospecção fitoquímicados dois extratos foi verificada a presença deflavonoides, taninos, saponinas, óleos essenciais e triterpenos. O A. microcarpumDfoio extrato que apresentou a menor concentração inibitória mínima, com valor de 0,78 mg/mLem cepas de S.aureus. Os extratos etanólicos das folhasapresentaram capacidade microbicida e antioxidante in vitro, o que sugereserem potenciais produtos para aelaboração de um fitoterápico. Artigo 3. Atividade antioxidante, análises bioquímicas e histopatológica de folhas de A. occidentale L. e A. microcarpum D. em camundongos knockout para interleucina 10. Para os ensaios foram utilizados camundongos knockout para interleucina 10, divididos em seis grupos: controle (G1) apenas água e ração; Anacardium occidentale (G2) água, ração e extrato; Anacardium microcarpum (G3) água, ração e extrato;paracetamol (G4) indução com paracetamol por oito dias; paracetamol+Anacardium occidentale (G5), tratados após injúria; paracetamol+Anacardium microcarpum (G6), tratados após injúria. Após serem anestesiadosfoi coletado o sangue retroorbital para contagem de leucócitos e para análises bioquímicas da alanina aminotransferase (ALT), aspartato transferase (AST), gama glutamil transferase (GGT), fosfatase alcalina (FAO), colesterol total e triglicerídeos. O macerado do fígado foi utilizado para as análises do estresse oxidativo a partir dos ensaios de malondialdeído (MDA), superóxido dismutase (SOD), catalase (CAT), glutationa e proteína carbonilada. Foi realizada a histopatologia do fígado, a partir da análise da presença ou ausência de alterações do núcleo; congestão sinusóide; esteatose; infiltrado inflamatório, colestase e vacuolização do citoplasma.A análise doestresse oxidativo em macerado de fígado apresentou melhores resultados nos camundongos tratados com Anacardium microcarpumD, na maioria das enzimas. Os camundongos tratados com os dois extratos, sem injúria por paracetamol e tratados com A. microcarpum após injúria, apresentaram resultados significativos para SOD e CAT, demonstrando eficiência na eliminação de espécies reativas de oxigênio. Destaca-se também o resultado da proteína carbonilada com redução efetiva nos grupos tratados com os dois extratos após injúria com paracetamol. Na análise histopatológica do fígado os camundongos tratados com os dois extratos apresentaram redução no infiltrado inflamatório e na colestase, demonstrando diminuição no processo inflamatório após a injúria pelo paracetamol. Os resultados confirmam a hipótese de atividade antioxidante das folhas do Anacardium occidentale L e do Anacardium microcarpum D, o que sugerea utilização das folhas para desenvolver um produto fitoterápico. / Mankind has always sought in plants alternative treatments to pathologies, and the knowledge and use of them, for many traditional communities, may be the only therapeutic option. Medicinal plants have been used for therapeutic purposes, in some cases without so much as the knowledge of their active ingredients and toxicity. On the other hand, there are several known actions performed by plant derivatives, being some: antiviral, antispasmodic, analgesic, antimicrobial, healing, expectorant, relaxing, antiseptic, respiratory, larvicidal, vermifuge and anti-inflammatory action. The increase of multiresistant bacteria stimulates interest in the search for new therapeutic alternatives with antimicrobial power. This has caused an emergence of research in the area, especially from plant resources. The Anacardium occidentale L (cashew) plant with a succulent pseudofruit, is characteristic of the tropical region of Brazil, and has pharmacological activities proven by the anti-inflammatory, astringent and antitumoral effects. Anacardium microcarpum (cajuí), present in the Cerrado region of the Amazon and in the Northeast, Center-West and Southeast regions of Brazil, presents a small pseudofruit and in its secondary metabolism produces phenolics, compounds with anti-inflammatory, antioxidant and antimicrobial activities. The aims of this study were to evaluate the antimicrobial activity of leaf extracts of Anacardium occidentale (cashew) and Anacardium microcarpum (cajuí) against multiresistant bacteria, and its antioxidant actions in vitro and in IL-10 knockout mice. The results were divided into three articles, which are composed of a review of the antimicrobial and antioxidant actions of pequi (Caryocar brasiliense), cashew (Anacardium occidentale) and cajuí (Anacardium microcarpum) and two original articles, one with antioxidant and antimicrobial in vitro and another with in vivo oxidative stress assays. Article 1. Antioxidant and antimicrobial activities of crude extracts and fractions of cashew (Anacardium occidentale L.), cajuí (Anacardium microcarpum) and pequi (Caryocar brasiliense C). A systematic review. A systematic review of the works that presented results of tests with extracts of the parts of the vegetal species mentioned above was elaborated. The databases Pubmed / Medline, Virtual Health Library (Lilacs and Scielo) and Science Direct were analyzed. Of a total of 425 articles, 32 were included, being 19 studies with Anacardium occidentale, 3 with Anacardium microcarpum and 10 with Caryocar brasiliense. The most used in vitro antioxidant capacity evaluation technique was DPPH and the most used microbiological capacity evaluation technique was the minimum inhibitory concentration and disk diffusion. Many articles were not found due to the limitations of the database search. Most of the tests described demonstrated potentially positive results to the antioxidant and antimicrobial actions when using the species studied. The results provide important data and perspectives for the use of natural products that can contribute to the treatment of various diseases, especially non-communicable diseases (NCD). Article 2. Phytochemical characterization, antimicrobial and antioxidant activities of Anacardium occidentale L and Anacardium microcarpum leaves, in vitro. Ethanolic extracts of A. occidentale L and A. microcarpum were prepared from crunched dry leaves for use in the trials. The DPPH and ABTS methodologies were used to evaluate the antioxidant power in vitro. Thin-layer chromatography was the technique of choice for phytochemical prospecting and HPLC (high performance liquid chromatography) to verify the presence of anacardic acids. The phenolics and total flavonoids of the extracts were quantified by spectrophotometry and linear regression. For the microbiological tests, the minimum inhibitory concentration in 96 microplates from hospital originated multiresistant strains was used. The ethanolic extracts of Anacardium occidentale and Anacardium microcarpum showed capture of similar DPPH and ABTS radicals and 2.34 and 1.64 mg GAE (galic acid equivalents) of total phenolics and total flavonoid values of 0.34 and 0, 29 mg RE (routine equivalents), respectively. The phytochemical prospection showed the presence of flavonoids, tannins, saponins, essential oils and triterpenes. The presence of the secondary flavonoid metabolites and tannins may favor the digestion of nutrients, the organic functioning of the body, activate the antioxidant capacity and modify the synthesis of eicosanoids, with anti-inflammatory response. A. microcarpum was the extract with the lowest minimum inhibitory concentration, with a value of 0.78 mg / mL in S. aureus strains. The ethanolic extracts of the leaves presented microbicidal and antioxidant capacity in vitro, which leads us to believe that they are potential products for the elaboration of a phytotherapeutic medicine. However, more studies are fundamental to uncover the gaps still existing in the knowledge about these plants. Article 3. Antioxidant activity of Anacardium occidentale D. and Anacardium microcarpum L. leaves in interleukin-10 knockout mice. For the tests, interleukin-10 knockout mice were used, divided into six groups (control, with injury and treated): control; paracetamol; paracetamol + Anacardium occidentale; paracetamol + Anacardium microcarpum; Anacardium occidentale and Anacardium microcarpum. Retroorbital blood was collected for leukocyte count and serum was separated for analysis of the biochemical profile of alanine aminotransferase (ALT), aspartate transferase (AST), gamma glutamyl transferase (GGT), alkaline phosphatase (FA), total cholesterol and triglycerides. Liver maceration was used for the analysis of oxidative stress from the malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione and carbonylated protein assays. The histopathological analysis of the liver was performed, observing the changes in the tissue in each group, analyzing the presence or absence of changes in the cytoplasm and nucleus; steatosis; inflammatory infiltrate and cholestasis. Oxidative stress in liver maceration showed better results in mice treated with Anacardium microcarpum in most of the enzymes. The result of the carbonylated protein with a value of 33.17 nmol / mg of the paracetamol group and 13.03 nmol/mg was also highlighted. The mice treated with the two extracts, without paracetamol injury and treated with A. microcarpum after injury, presented significant results for SOD and CAT, demonstrating efficiency in the elimination of reactive oxygen species. In the histopathological analysis of the liver, the mice treated with the two extracts presented reduction in the inflammatory infiltrate and cholestasis, demonstrating a decrease in the inflammatory process after the injury by paracetamol. The results confirm the hypothesis of antioxidant activity of the leaves of Anacardium occidentale and Anacardium microcarpum, which suggests, from more detailed studies, the use of leaves to develop a phytotherapeutic agent.