Estudo fitoquímico e avaliaçao das atividades alelopáticas e antimicrobianas das partes aéreas de Pterocaulon lorentzii Malme (Asteraceae)

Orientador : Obdulio Gomes MiguelCo-orientadora : Marilis Dallarmi MiguelDissertaçao (mestrado) - Universidade Federal do Paraná, Setor de Ciencias da Saúde, Programa de Pós-Graduaçao em Ciencias Farmaceuticas. Defesa: Curitiba, 2008Inclui bibliografiaÁrea de concentraçao: Insumos, medicamentos e correlato

Estudo das atividades biológicas in vitro da 5-(2,3-Dihidroxi-3-Metilbutilox)-6,7- Metilenedioxicumarina isolada de Pterocaulon lorentzii Malme E Pterocaulon alopecuroides DC. com ênfase na atividade anti-helmíntica em Nematoides gastrintestinais de ovinos

Orientador: Prof. Dr. Obdulio Gomes MiguelOrientadora: Profª Drª Marilis Dallarmi MiguelTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa: Curitiba, 06/09/213Bibliografia: f. 95-108Área de concentração: Insumos, medicamentos e correlatosResumo: Infecções gastrintestinais causadas por nematoides constituem um dos principais fatores limitantes para a ovinocultura no Brasil e em todo o mundo. A saúde dos rebanhos depende de um controle antiparasitário eficiente e a utilização de plantas medicinais como alternativa fitoterápica na medicina veterinária vêm ganhando espaço e se mostrando próspera. Tendo em vista os aspectos sociais, econômicos e ambientais este trabalho tem por objetivo estudar numa abordagem multidisciplinar a efetividade dos extratos, frações e da 5-(2,3-dihidroxi-3-metilbutiloxi)-6,7-metilenedioxicumarina isolada das espécies Pterocaulon alopecuroides DC. e Pterocaulon lorentzii Malme. Como foco buscou-se a efetividade destas espécies em ensaios antiparasitários in vitro contra nematoides gastrintestinais de ovinos além dos seus potenciais antioxidantes e da toxicidade. As partes aéreas secas e estabilizadas de ambas espécies foram submetidas a extração com etanol e particionadas com solventes com gradiente de polaridade crescente hexano-clorofórmio-acetato de etila em aparelho de soxhlet modificado. A partir da cromatografia líquida em coluna da fração clorofórmio e elucidação estrutural por RMN 1H e 13C foi identificada a cumarina 5-(2,3-dihidroxi-3- metilbutiloxi)-6,7-metilenedioxicumarina. Os extratos brutos, as frações e a cumarina foram submetidos a bioensaios in vitro para avaliação de toxicidade, atividade antioxidante, e atividade anti-helmíntica. Na avaliação da ecotoxicidade frente ao microcrustáceo Artemia salina as amostras testadas apresentaram atividade apenas em concentrações superiores a 1000?g/mL, consideradas não tóxicas neste ensaio. Da mesma forma, na avaliação da atividade hemolítica, tanto no teste em sangue de carneiro como em ágar sangue os extratos, as frações e a cumarina não apresentaram atividade. O potencial antioxidante foi avaliado por três metodologias. No teste de inibição do complexo fosfomolibdênio destacou-se a fração acetato de etila de P. alopecuroides que apresentou 36,4% de atividade em relação à vitamina C e 137,7% em relação à rutina. A fração acetato de etila de P. lorentzii também apresentou maior atividade em relação às demais amostras testadas ARR(%) de 101,7% em relação à vitamina C. No método de reação com o radical DPPH as frações acetato de etila de ambas espécies mostraram as melhores atividades, EC50 10,74 ?g/mL para P. alopecuroides e EC50 7,63 ?g/mL para P. lorentzii, em relação ao controle vitamina C (EC50 2,48 ?g/mL) e rutina (EC50 9,43 ?g/mL). As demais amostras, extrato bruto, fração hexano, fração clorofórmio e cumarina não apresentaram atividade significativa. No teste de reação ao ácido tiobarbitúrico (TBARS) as amostras mais expressivas foram os extratos brutos das duas espécies em relação ao controle positivo BHT cujo IA% 0,169 ± 0,0087. O extrato bruto de P. alopecuroides apresentou IA% 0,419 ± 0,0517 e o extrato bruto de P. lorentzii IA% 0,213 ± 0,0094. Já a 5-(2,3-dihidroxi-3-metilbutiloxi)-6,7- metilenedioxicumarina apresentou IA% 0,462 ± 0,0184. Na avaliação da atividade antihelmíntica pelo método da eclodibilidade de ovos de nematoides a cumarina apresentou atividade muito expressiva na inibição do desenvolvimento e eclosão dos ovos com EC50 3,2 ?g/mL, seguido das frações clorofórmio 17,2 ?g/mL (P. alopecuroides) e 9,4 ?g/mL (P. lorentzii) enquanto que as demais frações e extratos apresentaram atividades menores. Da mesma forma a cumarina apresentou maior atividade na inibição da migração de larvas de nematóides em ágar com EC50 3,1 ?g/mL e as frações clorofórmio 104,1?g/mL (P. alopecuroides) e 64,7 ?g/mL (P. lorentzii). Os resultados obtidos sugerem a aplicação dos extratos e frações de P. alopecuroides e P. lorentzii e a 5-(2,3-dihidroxi-3-metilbutiloxi)-6,7-metilenedioxicumarina no desenvolvimento de preparações farmacêuticas de uso veterinário.Abstract: Gastrointestinal infection caused by nematodes are the main limiting factor for sheep breeding in Brazil and around the world. Livestock healthiness depends on an efficient anthelmintic control and the use of medicinal plants as an herbal alternative in veterinary medicine is gaining space and showing prosperous. Considering the social, economic and environmental scope this work aims to study in a multidisciplinary approach, the effectiveness of extracts, fractions and 5-(2,3-dihydroxy-3-methylbutyloxy)-6,7- methylenedioxicoumarin isolated from Pterocaulon alopecuroides DC. and Pterocaulon lorenztii Malme species. This study was focused on the effectiveness of these two species in vitro anthelmintic assays against sheep gastrointestinal nematodes as well as their antioxidant and toxicity potential. The dried and stabilized aerial parts of both species were subjected to extraction with ethanol and partitioned with solvents of increasing polarity gradient hexane-chloroform-ethyl acetate in modified soxhlet apparatus. From the liquid column chromatography of chloroform fraction an the structure elucidation by NMR 1H and NMR 13H the 5-(2,3-dihydroxy-3-methylbutyloxy)- 6,7-methylenedioxicoumarin was identified. The crude extracts, fractions and the coumarin were submitted to in vitro bioassays to evaluate toxicity, antioxidant activity, and anthelmintic activity. In the ecotoxicity assessment against Artemia salina tested samples showed activity only at concentrations over 1000?g/mL considered non-toxic in this assay. Likewise, in the hemolytic activity assay test in sheep blood and blood agar, extracts, fractions and the coumarin showed no activity. The scavenging activity was evaluated by three methods. In the phosphomolybdenium complex inhibition test the ethyl acetate fraction of P. alopecuroides showed 36.4% activity compared to vitamin C and 137.7% compared to rutin. The ethyl acetate fraction of P. lorentzii also showed higher activity in comparison to other samples tested with ARR (%) 101.7% compared to vitamin C. In DPPH reaction methods ethyl acetate fractions of both species showed the best activities, EC50 10.74 mg/mL for P. alopecuroides and EC50 7.63 mg/mL for P. lorentzii compared to the control Vitamin C (EC50 2.48g/mL) and rutin (EC50 9.43 g/mL). The remaining samples, crude extract, hexane fraction, chloroform fraction and the coumarin showed no significant activity. The crude extracts of the two species showed the most significant results in thiobarbituric acid reaction test (TBARS) in relation to the positive control BHT (AI 0.169 ± 0.0087%). The crude extract of P. alopecuroides presented IA 0.419% ± 0.0517 and the crude extract of P. lorentzii IA 0.213% ± 0.0094. The 5-(2,3-dihydroxy-3-methylbutyloxy)-6,7-methylenedioxicoumarin showed0020cIA 0.462% ± 0.0184. In the evaluation of anthelmintic activity by the egg hatch test (EHT) the coumarin showed very significant activity in inhibiting sheep nematoid egg development hatching with EC50 3.2 mg/mL, followed by chloroform fractions 17.2?g/mL (P. alopecuroides) and 9.4 mg/mL (P. lorentzii) while the other fractions and extracts showed lower activity. As well as in the EHT the same coumarin showed higher effectiveness in inhibiting nematode larvae migration in agar with a EC50 3.1 g/mL followed by the chloroform fractions 104.1g/mL (P. alopecuroides) and 64.7 mg/mL (P. lorentzii). The obtained results suggest the application of extracts and fractions of P. alopecuroides and P. lorentzii and 5-(2,3-dihydroxy-3-methylbutyloxy)-6,7- methylenedioxicoumarin in the development of pharmaceutical preparations for veterinary use

Differential, histochemical and immunohistochemical changes in rat hepatocytes after isoflurane or sevoflurane exposure.

Differential, histochemical and immunohistochemical changes were observed in hepatocytes from immediately to 7 days after isoflurane or sevoflurane exposure (at H 0 to on Day 7) to study the process of development and recovery in anesthetic-induced hepatic injury. A total of 570 7-week-old male Sprague-Dawley rats with or without phenobarbital treatment were exposed to isoflurane or sevoflurane in 100%, 21%, or 10% oxygen, or to 10% oxygen alone for 2h. In phenobarbital-treated rats, hepatocytes both with and without anesthetic exposure markedly changed in 10% oxygen at H 0. Glycogen and ribosomal ribonucleic acid (rRNA) disappeared at H 0 and at H 6, respectively, and at H 6, AST levels in the blood rose. From H 6 to Day 1, necrosis developed more markedly and widely in zone 3 hepatocytes exposed to anesthetics in 10% oxygen than in those exposed to oxygen alone. All degenerated tissues had returned to normal levels by day 7. Recovery of the hepatolobular structure may be attributed to rearrangement of remaining hepatocytes in the portal vein area. Both the disappearance of glycogen and rRNA and the increase in blood AST levels after exposure to isoflurane or sevoflurane are considered to be factors contributing to the induction of necrosis around the central vein. The grade of isoflurane-induced hepatic injury was found to be significantly higher than that of sevoflurane.</p

Pediatric eosinophilic gastroenteritis

Eosinophilic gastrointestinal disorders are chronic inflammatory diseases in which eosinophils highly infiltrate into gastrointestinal tissue, resulting in gastrointestinal dysfunction. Here, we report a case of pediatric eosinophilic gastroenteritis (EGE). A 7-year-old boy with multiple food allergies (cow milk, hen's egg, fish, shellfish, and chicken) was admitted to our hospital because of continuous abdominal pain and vomiting. His soy allergy had been diagnosed to have oral tolerance based on an oral food challenge at the age of 6 years. He was diagnosed with EGE based on biopsy findings showing eosinophilic infiltration (>20 eosinophils per high-power field) into the gastrointestinal mucosa. A diet eliminating soy, wheat, beef, pork, rice, and sesame in addition to the food that had already been eliminated and oral corticosteroids improved his symptoms and peripheral eosinophilia. A relapse of both abdominal pain and peripheral eosinophilia after the reintroduction of soy or pork identified them as foods causative of EGE. This report highlights the utility of elimination diets in improving EGE symptoms and the subsequent reintroduction of offending foods in identifying causative foods. Furthermore, EGE onset should be considered when introducing potentially allergic food in the management of food allergy

Comparative Study on Epstein-Barr Virus-Positive Mucocutaneous Ulcer and Methotrexate-Associated Lymphoproliferative Disorders Developed in the Oral Mucosa: A Case Series of 10 Patients and Literature Review

Methotrexate-associated lymphoproliferative disorder (MTX-LPD) is an iatrogenic immunodeficiency-associated lymphoproliferative disorder that occurs mainly with MTX use. This disorder has been associated with Epstein-Barr virus (EBV) infection. In 2017, the WHO newly defined the disease concept of EBV-positive mucocutaneous ulcer (EBV-MCU) as a good-prognosis EBV-related disease. Here, we report 10 cases of MTX-LPD or EBV-MCU in the oral mucosa. This retrospective, observational study was conducted with MTX-LPD or EBV-MCU in the oral mucosa patients who visited us during the nine year period from 2012 to 2021. We gathered the basic information, underlying disease, histopathological evaluation, treatment and prognosis for the subjects. All were being treated with MTX for rheumatoid arthritis. EBV infection was positive in all cases by immunohistochemistry. A complete or partial response was obtained in all cases with the withdrawal of MTX. Our results suggests that the most common risk factor for developing EBV-MCU is the use of immunosuppressive drugs. The most common site of onset is the oral mucosa, which may be attributed to the mode of EBV infection and the high incidence of chronic irritation of the oral mucosa. A small number of patients had been diagnosed with MTX-LPD, but we consider that these cases were EBV-MCU based on our study

Comparison of Anti-Inflammatory Analgesics for Mechanical Stress-induced Inflammation in a Human Synovial Sarcoma Cell Line

Osteoarthritis is a complicated clinical condition affected by age, mechanical stress, cartilage hypertrophy, cytokines, and genetic predisposition. In this study, we compared the effects of various anti-inflammatory analgesics on mechanical stress-induced inflammation in a synovial sarcoma cell line （SW982 cells）. SW982 cells exposed to mechanical stress by shaking with hydroxyapatite-simulating bone chips were treated with acetaminophen, ketoprofen, triamcinolone acetonide, celecoxib, or neurotrophin for 48hr. The expression of integrin α5β1 receptor, observed in fibroblasts and synovium, was evaluated. Levels of the transcription factor, nuclear factor-κB, the inflammatory cytokine, tumor necrosis factor-α, the proteolytic enzyme, matrix metalloproteinase-3, and prostaglandin E2, which is associated with pain and arachidonate cascade product levels, were measured by ELISA. The expression of integrin α5β1 was significantly increased by mechanical stress. Activation of nuclear factor-κB by mechanical stress was significantly suppressed by celecoxib only. Mechanical stress-induced increases in tumor necrosis factor-α and matrix metalloproteinase-3 levels were significantly suppressed by acetaminophen, triamcinolone acetonide, and neurotrophin. The mechanical stress-induced increase in prostaglandin E2 levels was significantly suppressed by acetaminophen, ketoprofen, and celecoxib. SW982 exposed to mechanical stress is proposed as a model for arthritis, and indeed, the expression of integrin α5β1, a membrane receptor protein that binds to fibronectin and the extracellular matrix, and is involved in cell proliferation, differentiation, and neovascularization in osteoarthritis, was significantly upregulated. Following evaluation using this model, acetaminophen was found to possess anti-inflammatory, analgesic, and joint-destruction suppression properties. This drug may, therefore, have applications in the treatment of mechanical stress-induced inflammation

Dispersion-Flocculation Behavior of Fine Lead Particles in an Organic Solvent

Methyl ethyl ketone (MEK) is a good solvent for polyvinyl chloride (PVC) and it has been proposed for use in PVC recycling. In the recycling process, fine particles of 3PbOÁPbSO 4 ÁH 2 O, used as a thermal stabilizer in PVC products, are dispersed and not dissolved in the solvent. To establish methods for removing of 3PbOÁPbSO 4 ÁH 2 O particles from the solvent, factors affecting the dispersion-flocculation behavior of the particles in MEK were investigated. The zeta potential and particle distribution of 3PbOÁPbSO 4 ÁH 2 O particles in MEK solutions containing known amounts of H 2 O were measured. Above 5 vol%H 2 O in MEK solutions, the zeta potential of 3PbOÁPbSO 4 ÁH 2 O particles approached zero and the flocculation of particles was achieved. In addition, it was found that Pb 2þ and Cl À affect the zeta potential of the particles. These results indicate that the dispersion-flocculation behavior of lead particles can be influenced by the concentration of H 2 O, Pb 2þ , and Cl À in MEK

Prediction of acid mine drainage formation and zinc migration in the tailings dam of a closed mine, and possible countermeasures

Acid mine drainage (AMD), the very acidic and highly contaminated leachate generated in closed/abandoned mines, is commonly managed by neutralization to raise the pH and precipitate most of the heavy metals. Although effective, this approach does not generate any product of economic value, so it is very costly and unsustainable in the long-term. Unfortunately, there are currently no effective alternatives to neutralization, and one way to improve the sustainability of this process is to reduce the volume of AMD generated and/or the concentration of heavy metals. The tailings dam investigated in this study is located in northern Hokkaido, Japan. Detailed characterization of borehole core samples showed that even after almost 40 years of exposure to the environment, the tailings still contain pyrite (FeS2) and substantial amounts of copper (Cu) and zinc (Zn). Reactive-transport modeling using Visual MODFLOW predicted that AMD quality would likely continue to deteriorate with time and that treatment should be continued for at least 1,000 years. The model also predicted that a barrier with low permeability installed downstream of the tailings dam or ground sealing techniques for recharge reduction could lower the volume of AMD and concentration of Zn from the site