Impact of an educational hands-on project on the antimicrobial, antitumor and anti-inflammatory properties of plants on Portuguese students' awareness, knowledge, and competences

Promoting environmental and health education is crucial to allow students to make conscious decisions based on scientific criteria. The study is based on the outcomes of an Educational Project implemented with Portuguese students and consisted of several activities, exploring pre-existent Scientific Gardens at the School, aiming to investigate the antibacterial, antitumor and anti-inflammatory properties of plant extracts, with posterior incorporation in soaps and creams. A logo and a webpage were also created. The effectiveness of the project was assessed via the application of a questionnaire (pre- and post-test) and observations of the participants in terms of engagement and interaction with all individuals involved in the project. This project increased the knowledge about autochthonous plants and the potential medical properties of the corresponding plant extracts and increased the awareness about the correct design of scientific experiments and the importance of the use of experimental models of disease. The students regarded their experiences as exciting and valuable and believed that the project helped to improve their understanding and increase their interest in these subjects and in science in general. This study emphasizes the importance of raising students’ awareness on the valorization of autochthonous plants and exploitation of their medicinal properties.

Evaluation of antinociceptive and anti-inflammatory activities of the topical preparation of Cipura paludosa (Iridaceae)

Alho do mato (Cipura paludosa, Iridaceae) is a medicinal plant found in the Amazon rain forest, North of Brazil. It has been used to treat algic, inflammatory and infectious processes. The aim of this study was to evaluate the anti-inflammatory and antinociceptive action of the crude Cipura paludosa ethanolic extract at concentrations ranging between 2.0 and 4.0% in Oil and Water cream formulations for topical use. The physical-chemical stability of the formulations was monitored over a six-month period with the use of accelerated stability tests. In order to evaluate the anti-inflammatory and antinociceptive activities, we used a paw edema test induced by carrageenan and a formalin test, respectively. The paw edema test showed that there was a statistical difference in the control group in relation to the treatments. The formalin test did not confirm antinociceptive action of the treatments with the extract in the early phase of the test. However, statistical difference was confirmed for the treatments in relation to the control in the late phase. The antinociceptive and anti-inflammatory activities of Cipura paludosa preparations, as demonstrated in the results, at least partially support the ethno-medical uses of this plant.

The contribution of phenolic acids to the anti-inflammatory activity of mushrooms: Screening in phenolic extracts, individual parent molecules and synthesized glucuronated and methylated derivatives

In the present study, the ethanolic extracts of fourteen edible mushrooms were investigated for their anti-inflammatory potential in LPS (lipopolysaccharide) activated RAW 264.7 macrophages. Furthermore the extracts were chemically characterized in terms of phenolic acids and related compounds. The identified molecules (p-hydroxybenzoic, p-coumaric and cinnamic acids) and their glucuronated and methylated derivatives obtained by chemical synthesis were also evaluated for the same bioactivity, in order to establish structure-activity relationships and to comprehend the effects of in vivo metabolism reactions in the activity of the compounds. The extracts of Pleurotus ostreatus, Macrolepiota procera, Boletus impolitus and Agaricus bisporus revealed the strongest anti-inflammatory potential (EC50 values 96 ± 1 to 190 ± 6 µg/mL, and also the highest concentration of cinnamic acid (656 to 156 µg/g), which was also the individual compound with the highest anti-inflammatory activity. The derivatives of p-coumaric acid revealed the strongest properties, specially the derivative methylated in the carboxylic group (CoA-M1) that exhibited similar activity to the one showed by dexamethaxone used as anti-inflammatory standard; by contrast, the derivatives of p-hydroxybenzoic revealed the lowest inhibition of NO production. All in all, whereas the conjugation reactions change the chemical structure of phenolic acids and may increase or decrease their activity, the glucuronated and methylated derivatives of the studied compounds are still displaying anti-inflammatory activity.

Barramento delgado anti carbonatação

Dissertação de mestrado integrado em Engenharia Civil

MET is required for the recruitment of anti-tumoural neutrophils

Mutations or amplification of the MET proto-oncogene are involved in the pathogenesis of several tumours, which rely on the constitutive engagement of this pathway for their growth and survival. However, MET is expressed not only by cancer cells but also by tumour-associated stromal cells, although its precise role in this compartment is not well characterized. Here we show that MET is required for neutrophil chemoattraction and cytotoxicity in response to its ligand hepatocyte growth factor (HGF). Met deletion in mouse neutrophils enhances tumour growth and metastasis. This phenotype correlates with reduced neutrophil infiltration to both the primary tumour and metastatic sites. Similarly, Met is necessary for neutrophil transudation during colitis, skin rash or peritonitis. Mechanistically, Met is induced by tumour-derived tumour necrosis factor (TNF)-a or other inflammatory stimuli in both mouse and human neutrophils. This induction is instrumental for neutrophil transmigration across an activated endothelium and for inducible nitric oxide synthase production upon HGF stimulation. Consequently, HGF/MET-dependent nitric oxide release by neutrophils promotes cancer cell killing, which abates tumour growth and metastasis. After systemic administration of a MET kinase inhibitor, we prove that the therapeutic benefit of MET targeting in cancer cells is partly countered by the pro-tumoural effect arising from MET blockade in neutrophils. Our work identifies an unprecedented role of MET in neutrophils, suggests a potential 'Achilles' heel' of MET-targeted therapies in cancer, and supports the rationale for evaluating anti-MET drugs in certain inflammatory diseases.

Tratamento anti-hipertensivo. Prescrição e custo de medicamentos. Pesquisa em hospital terciário

OBJETIVO: Verificar os medicamentos anti-hipertensivos mais utilizados por pacientes que procuram atendimento em hospital público terciário, avaliando o impacto das diretrizes de atendimento (consensos) e custo de aquisição. MÉTODOS: Foram selecionados 141 pacientes (101 do sexo feminino) de 40 a 72 (média 53,3) anos, que procuraram de forma espontânea, atendimento em hospital terciário, com diagnóstico prévio de hipertensão arterial feito por médico e ausência de queixas relacionadas ao aparelho cardiovascular. RESULTADOS: Verificou-se que 75,9% (n=107) estavam em uso diário de anti-hipertensivos, sendo 60,7% (n=86) em monoterapia e os demais em terapia mista. Os medicamentos mais empregados em monoterapia eram: tiazídicos, metildopa, inibidores da ECA, bloqueadores de canal de cálcio e betabloqueadores. A combinação com tiazídicos (26,3% do total) seguiu a mesma preferência. O segundo medicamento mais prescrito, metildopa, era o de maior custo. Metade dos pacientes adquiriu os medicamentos por compra direta. CONCLUSÃO: Observou-se maior utilização de anti-hipertensivos de alto custo, conduta discordante das principais diretrizes das sociedades médicas, sobretudo do V-JNC, que preconizou tiazídicos e betabloqueadores, como anti-hipertensivos de primeira escolha em hipertensos sem complicações ou condições associadas.

Structural and enzymatic characterization of ABgp46, a novel phage endolysin with broad anti-Gram-negative bacterial activity

The present study demonstrates the antibacterial potential of a phage endolysin against Gram-negative pathogens, particularly against multidrug resistant strains of Acinetobacter baumannii. We have cloned, heterologously expressed and characterized a novel endolysin (ABgp46) from Acinetobacter phage vb_AbaP_CEB1 and tested its antibacterial activity against several multidrug-resistant A. baumannii strains. LC-MS revealed that ABgp46 is an N-acetylmuramidase, that is also active over a broad pH range (4.0-10.0) and temperatures up to 50°C. Interestingly, ABgp46 has intrinsic and specific anti-A. baumannii activity, reducing multidrug resistant strains by up to 2 logs within 2 hours. By combining ABgp46 with several organic acids that act as outer membrane permeabilizing agents, it is possible to increase and broaden antibacterial activity to include other Gram-negative bacterial pathogens. In the presence of citric and malic acid, ABgp46 reduces A. baumannii below the detection limit (> 5 log) and more than 4 logs P. aeruginosa and Salmonella Typhimurium strains. Overall, this globular endolysin exhibits a broad and high activity against Gram-negative pathogens, that can be enhanced in presence of citric and malic acid, and be used in human and veterinary medicine.

Anti-inflammatory and antinociceptive activities of Bauhinia monandra leaf lectin

A galactose-specific lectin from Bauhinia monandra leaves (BmoLL) have been purified through ammonium sulphate fractionation followed by guar gel affinity chromatography column. This study aimed to evaluate the potential anti-inflammatory and antinociceptive activity of pure BmoLL in mice. Anti-inflammatory activity was evaluated by 1% carrageenan-induced inflammation in mice treated with BmoLL. Acetic acid-induced abdominal writhing and hot plate methods evaluated antinociceptive activity. BmoLL significantly inhibited the carrageenan-induced paw edema by 47% (30 mg/kg) and 60.5% (60 mg/kg); acetylsalicylic acid (ASA, 100 mg/kg) showed inhibition of 70.5%, in comparison to controls. Leukocyte migration, an immune response to the inflammation process, was significantly reduced in presence of BmoLL; in mice treated with \ASA\ the decrease in leukocyte migration was similar to 15 mg/kg of the lectin. BmoLL at doses of 15, 30 and 60 mg/kg significantly reduced the number of animal contortions by 43.1, 50.1 and 71.3%, respectively.BmoLL leukocyte migration was significantly reduced; in mice treated with \ASA\ the decrease in leukocyte migration was similar to 15 mg/kg of the lectin. BmoLL at doses of 15, 30 and 60 mg/kg significantly reduced the number of animal contortions by 43.1, 50.1 and 71.3%, respectively. The lectin (30 and 60 mg/kg) showed a significant effect in the hot plate assay. BmoLL anti-inflammatory and antinociceptive effects were dose-dependent. The search for new and natural compounds, with minimal side effects, to control pain and inflammation, is constantly increasing. BmoLL has great potential as a natural anti-inflamatory product that can be explored for pharmacological purposes.

Kawasaki disease: involution of giant coronary aneurysms after prolonged anti-inflammatory treatment

The patient is a 5-year-old male with Kawasaki disease, whose involution of giant aneurysms of the left coronary arteries was surprising after a prolonged period of treatment, which lasted 80 uninterrupted days and comprised anti-inflammatory drugs associated with anticoagulation agents. The distal diameters of the anterior interventricular, the diagonal, and the circumflex arteries normalized by the end of the treatment. A residual giant aneurysm localized at the beginning of the anterior interventricular artery did not cause ischemia. Continuation of the medication for a prolonged period was recommended.

Anticuerpos anti-glicoesfingolípidos

Los glicoesfingolípidos (GSL) son moléculas anfipáticas, componentes de la membrana plasmática. La porción hidrofóbica, formada por un aminoalcohol de cadena larga N-acilado (ceramida), se inserta en la bicapa lipídica; mientras que la porción hidrofílica, muy variable y expuesta al medio extracelular, está compuesta por uno o varios azúcares. Como grupo son moléculas ubicuas, aunque existe especificidad en el contenido de GSL de ciertos tipos celulares y de tejidos. Estas características hicieron que estos compuestos fueran considerados como moléculas potencialmente claves en el reconocimiento celular. GSL han sido implicados en procesos de interacción célula-célula y de reconocimiento celular por toxinas y anticuerpos. GSL portando determinadas estructuras en el oligosacárido han sido identificados como antígenos de grupo sanguíneo y antígenos específicos de tumores. Más recientemente, GSL han sido propuestos como antígenos en neuropatías autoinmunes asociadas y no asociadas a gamopatías. Anticuerpos anti-GSL dirigidos contra heteroantígenos, tales como el antígeno de Forsmann y GSL de insectos o contra autoantígenos, como gangliósidos, son parte del repertorio normal de anticuerpos en el ser humano. Objetivos Generales: Nuestro laboratorio tiene como línea de investigación el estudio de anticuerpos anti-GSL en procesos patológicos. Estamos interesados tanto en aspectos básicos, como caracterización, origen y rol patológico en la enfermedad, como en aplicados: diagnóstico y monitoreo de terapias.

AIDS na infância: acometimento cardíaco com e sem a terapia anti-retroviral tríplice combinada

OBJETIVO: Descrever a prevalência de alterações cardíacas ao ecocardiograma em crianças com AIDS acompanhadas em serviço de referência aos 18±6 meses do diagnóstico confirmado de AIDS. MÉTODOS: Estudo transversal, com corte aos 18±6 meses do diagnóstico de AIDS. Incluídas 93 crianças com diagnóstico confirmado de AIDS por transmissão vertical, sem doença maligna, que, na avaliação cardiológica, realizaram ecocardiograma (eco). De forma exploratória avaliaram-se as alterações cardíacas nos pacientes sem uso (G1) e com uso (G2) de terapia combinada anti-retroviral. RESULTADOS: Quando do diagnóstico de AIDS, as crianças tinham em média 3,07 anos e 50,50% eram do sexo feminino. Esquema de terapia combinado com anti-retrovirais foi utilizado por 47 pacientes (G2). O acometimento cardíaco esteve presente em 40 crianças (43,00%). A presença de disfunção ventricular esquerda (G1:39,10%;G2:10,60%) e o aumento isolado de ventrículo esquerdo (G1:6,60%;G2:14,90%) foram os achados mais freqüentes. Observou-se associação significativa entre os grupos sem e com terapia anti-retroviral combinada quanto à presença de disfunção ventricular esquerda (RP=3,42; [1,41-8,26]; p =0,02) e de desnutrição (RP=1,79; [1,00-3,20]; p=0,04). CONCLUSÃO: O acometimento cardíaco foi freqüente nas crianças com AIDS, sendo a disfunção ventricular esquerda a alteração mais observada ao ecocardiograma. Houve diferença estatisticamente significativa entre os grupos com e sem tratamento tríplice combinado quanto à presença de disfunção ventricular esquerda e de desnutrição.

Neuropatías asociadas a anticuerpos anti-GM1. Anti-GM1 antibodies associated neuropathies.

Durante varios años estudiamos los anticuerpos presentes en pacientes con neuropatías e individuos normales. Recientemente hemos implementado un modelo animal de neuropatía inducido por inmunización con GM1, y nos proponemos confirmar en este modelo las observaciones realizadas en pacientes. Se caracterizará la respuesta inmune humoral durante la inducción de la enfermedad, y la posible existencia de un mecanismo regulatorio mediado por anticuerpos bloqueantes de anticuerpos anti-GM1. Además, el modelo nos permitirá ensayar diferentes estrategias terapéutica tales como plasmaféresis específica, bloqueo de anticuerpos por antígenos solubles e inducción de anticuerpos bloqueantes.

Tecnología de igy anti-escherichia coli aplicada mediante nanotubos sobre cultivos de enterocitos porcinos. Anti- escherichia coli igy technology applied through nanotubes on porcine enterocyte cultures.

La diarrea neonatal representa uno de los problemas sanitarios de mayor relevancia en las primeras semanas de vida del cerdo. Provoca importantes pérdidas económicas por morbilidad y mortalidad. El cultivo de enterocitos primarios representa una herramienta valiosa para el estudio de patologías causadas por agentes infecciosos que afectan la integridad del epitelio intestinal. La producción de anticuerpos extraídos a partir de la yema de huevo de gallinas inmunizadas (IgY), es una tecnología innovadora, que ha mostrado ser protectiva contra diarreas causadas por agentes víricos y bacterianos. La nanotecnología permite mejorar la eficiencia en la administración de distintas drogas. Los nanotubos de carbono han ganado una enorme popularidad por sus propiedades y aplicaciones únicas. La investigación sobre los aspectos toxicológicos de estas nanopartículas es escasa. Una vez dentro de la célula, las nanopartículas pueden inducir estrés oxidativo intracelular por perturbar el equilibrio oxidativo. Las hipótesis de trabajo es: La administración de IgY anti-Escherichia coli a través de nanotubos protegerá in vitro e in vivo a los enterocitos de una infección por E. coli previniendo la diarrea neonatal porcina. Los objetivos del trabajo son: Evaluar la protección por un anticuerpo aviario IgY anti-E. coli aplicado mediante nanotubos de carbono a cultivo de enterocitos porcinos primarios sometidos a una post-infección con E. coli; Analizar los efectos secundarios de los nanotubos con IgY anti-E coli en la citotoxicidad, el balance oxidativo y la apoptosis de los enterocitos porcinos cultivados in vitro y Evaluar la acción terapeútica de la IgY anti-E coli aplicada a porcinos y efectos secundarios de la administración con nanotubos. Se implementará un diseño experimental in vitro con diferentes grupos de cultivos con nanotubos, con IgY anti-E. coli e inespecifica y con exposición a E. coli. Se realizará cultivo de enterocitos porcinos primarios con una técnica de disgregación enzimática con colagenasa según protocolo de Bader et al. (2000). Se evaluará la viabilidad por la prueba de azul tripan. Para la obtención del anticuerpo anti-E. coli aviario se aplicarán un total de 3 dosis de E. coli (109 UFC/ml de adyuvante) a gallinas Legorhn en condiciones fisiológicas. Se recolectarán los huevos diariamente. Se purificará la IgY según método de Polson et al. (1985) utilizando PEG 6000. La concentración de IgY se medirá por ELISA de alta sensibilidad. La IgY será incorporada a nanotubos según protocolo de Acevedo et al. 2006. Para analizar los posibles efectos secundarios de los nanotubos se evaluará: 1. Citotoxicidad por técnica de MTT 2. Estrés oxidativo por técnica de TBARS y 3. Apoptosis por técnica de TUNEL.Además, se implementará un diseño experimental in vivo para probar la acción terapeútica de este nutraceútico aplicados a lechones destetados y los efectos secundarios de la administración con nanotubos. Se realizará un cultivo de enterocitos de lechones que previamente fueron tratados con la IgY anti-E. coli administrada mediante nanotubos y efectuarán las técnicas descriptas anteriormente. Los resultados esperados son: Elaboración de un Ac aviario IgY anti-E. coli para prevenir infección de enterocitos, Profundización en el conocimiento acerca de los efectos citotóxicos de los nanotubos de carbono multilamelares, Generación de tratamiento alternativo para enfermedades entéricas porcinas.