Influência de diferentes extratos aquosos de plantas medicinais no desenvolvimento de Colletotrichum gloeosporioides e de Fusarium moniliforme / Influence of different aqueous extracts of medicinal plants in the development of Colletotrichum gloeosporioides and Fusarium moniliforme

O objetivo deste trabalho foi avaliar a influência dos extratos aquosos das plantas medicinais alecrim, alho, cravo-da-índia, sálvia, capim-limão, orégano ou pimenta-do-reino no desenvolvimento in vitro de Colletotrichum gloeosporioides e de Fusarium moniliforme. Os extratos foram obtidos pela infusão de 60 g de cada planta medicinal em 200 mL de água fervente. Cada extrato aquoso foi fracionado em concentrações de 0, 5, 10 e 20% (p:v) e incorporado ao meio de cultivo BDA (batata-dextrose-ágar) antes da esterilização em autoclave. Posteriormente, um disco de 8 mm de diâmetro de micélio fúngico de cada patógeno foi transferido para o centro de placas de Petri. Após 24, 48 e 96 horas de incubação em câmara de crescimento a 22 ± 2 ºC e fotoperíodo de 12 horas avaliou-se o crescimento micelial de F. moniliforme e de C. gloesporioides. No último período de incubação, também se quantificou o número de conídios de cada fungo. Para o teste de germinação adicionou-se nas cavidades de placas de teste Elisa, uma alíquota de 40 µL de cada extrato nas concentrações de 0, 5, 10 e 20%, e outra alíquota, da suspensão de conídios de cada patógeno. Após 24 horas a 22 ± 2 ºC, no escuro, a germinação dos conídios foi paralisada com a adição de 20 µL de lactofenol; avaliou-se então a porcentagem de germinação de conídios. Os experimentos foram conduzidos no delineamento inteiramente casualizado em esquema fatorial 7 x 4 (extratos de plantas medicinais x concentrações) com quatro repetições. Para ambos os patógenos o extrato aquoso de alho e cravo-da-índia apresentaram maior ou total inibição do crescimento micelial, respectivamente, quando comparado com os demais extratos. Para C. gloeosporioides, o extrato de cravo-da-índia apresentou menor número de conídios em todas as concentrações testadas, e para o extrato de alho a 20%, também não foi observada a germinação de conídios. O extrato de alho foi eficiente em reduzir o número e a germinação dos conídios de F. moniliforme na concentração de 20%. Os extratos de alecrim, cravo-da-índia, orégano e pimenta-do-reino, nas maiores concentrações, tiveram efeito positivo na redução da produção de conídios deste mesmo fungo.

The objective of this study was to evaluate the influence of aqueous extracts of the medicinal plants rosemary, garlic, clove, sage, lemongrass, oregano and black pepper in the in vitro development of Colletotrichum gloeosporioides and Fusarium moniliforme. The extracts were obtained by infusing 60 g of each medicinal plant in 200 mL of boiling water. Each aqueous extract was fractionated in the concentrations of 0, 5, 10 and 20% (w:v) and incorporated into the PDA (potato dextrose agar) culture medium before sterilization by autoclaving. Later, an 8 mm diameter disc of each pathogen mycelium was transferred to the center of the Petri dishes. After 24, 48 and 96 hours of incubation in a growth chamber at 22 ± 2 ºC and a photoperiod of 12 hours, we evaluated the mycelial growth of F. moniliforme, and C. gloesporioides. In the last period of incubation, we quantified the production of conidia of each fungus. For the germination test, we added, into the wells of an ELISA test plates, a 40 µL aliquot of each extract at the concentrations of 0, 5, 10 and 20% and another aliquot of a suspension of conidia of each pathogen. After 24 hours at 22 ± 2 ºC in the dark, the germination of the fungi was stopped with the addition of 20 µL of lactophenol. Then, we evaluated the germination of conidia. The experiments followed a completely randomized 7 x 4 factorial design (medicinal plants x concentrations) with four replications. For both pathogens, the aqueous extract of garlic and clove showed a greater or total inhibition of the mycelial growth, when compared to the other extracts. For the C. gloeosporioides, the clove extract showed a lower number of conidia at all concentrations tested, and the garlic extract at 20% showed not conidial germination. The garlic extract was efficient to reduce the conidial number and germination of F. moniliforme at 20%. Extracts of rosemary, clove, oregano and black pepper, in the highest concentrations, had positive effect in reducing the production of spores of the same fungus.

Protocatechuic acid alkyl esters: hydrophobicity as a determinant factor for inhibition of NADPH oxidase.

This study presents the increased efficiency of NADPH oxidase inhibition produced by esterification of protocatechuic acid (P0). Alkyl esters bearing chain lengths of 4 (P4), 7 (P7) and 10 (P10) carbons were synthesized and their oxidation potential, hydrophobicity, antiradical activity, inhibition of superoxide anion (O2°(-)), and the abilities to affect hypochlorous acid (HOCl) production by leukocytes and inhibit myeloperoxidase (MPO) chlorinating activity were studied. The increased hydrophobicity (logP, 0.81-4.82) of the esters was not correlated with a significant alteration in their oxidation potential (0.222-0.298 V). However, except for P10, the esters were ~ 2-fold more effective than the acid precursor for the scavenging of DPPH and peroxyl radicals. The esters were strong inhibitors of O2°(-) released by activated neutrophils (PMNs) and peripheral blood mononuclear cells (PBMCs). A correlation was found between the carbon chain length and the relative inhibitory potency. P7, the most active ester, was ~ 10-fold more efficient as NADPH oxidase inhibitor than apocynin. The esters strongly inhibited the release of HOCl by PMNs, which was a consequence of the inhibition of NADPH oxidase activity in these cells. In conclusion, as effective inhibitors of NADPH oxidase, the esters of protocatechuic acid are promising drugs for treatment of chronic inflammatory diseases. Moreover, this is the first demonstration that, besides the redox active moiety, the hydrophobicity can also be a determinant factor for the design of NADPH oxidase inhibitors.

Extrato de alho no controle in vitro e in vivo da antracnose da videira / Garlic extract in the in vitro and in vivo control of anthracnose of grapevine

O objetivo deste trabalho foi avaliar a eficiência in vitro e in vivo do extrato bruto de alho no controle da antracnose da videira (Elsinoe ampelina). No primeiro experimento in vitro, adicionaram-se doses de 0, 5, 10, 15, 20, 25 ou 30 mL L-1 de extrato bruto de alho em meio batata-dextrose-ágar (BDA) antes da esterilização em autoclave e em meio fundente. Após 3, 5, 7 e 9 dias de incubação a 24 ± 2ºC e fotoperíodo de 16 horas, mensurou-se o crescimento micelial de E. ampelina. No segundo experimento in vitro, repetiu-se a metodologia de adição de extrato em meio fundente acrescentando 2,5 mL L-1 de óleo vegetal e uma testemunha absoluta somente com BDA. Avaliação da germinação de E. ampelina foi realizada após duas horas e após quatro horas de incubação a 24ºC e luz constante. O delineamento experimental utilizado para os experimentos in vitro foi inteiramente casualizado, com quatro repetições e parcela experimental constituída por uma placa de Petri. No experimento a campo, logo após a poda da videira cv. Isabel pulverizou-se semanalmente, sobre as folhas da planta, as doses de extrato bruto de alho acrescida do óleo, exceto na testemunha absoluta (sem tratamento). A partir dos primeiros sintomas da antracnose da videira, avaliou-se a severidade que foi expressa em área abaixo da curva do progresso da doença (AACPD). O delineamento foi em blocos ao acaso com cinco repetições. Constatou-se que o extrato bruto de alho reduziu o crescimento micelial do patógeno, principalmente ao adicioná-lo em meio de cultura, antes da esterilização, quando expressou o máximo potencial antifúngico. Ao adicionar o óleo vegetal às doses de extrato, constatou-se inibição total nas doses de 25 ou 30 mL L-1. Este efeito aditivo entre esses compostos também foi constato no teste de germinação de E. ampelina. Em condições de campo, o extrato bruto de alho reduziu a AACPD em 83,59% na dose de 25 mL L-1.

The aim of this study was to evaluate the in vitro and in vivo efficacy of the crude extract of garlic in controlling anthracnose of grapevine (Elsinoe ampelina). In the first experiment in vitro, 0, 5, 10, 15, 20, 25 or 30 mL L-1 crude extract of garlic were added to potato-dextrose-agar medium (PDA) before autoclaving for sterilization and to melting media. After 3, 5, 7 and 9 days of incubation at 24 ± 2ºC and photoperiod of 16 hours, the mycelial growth of E. ampelina was measured. In the second experiment in vitro, we repeated the methodology of adding the extract to the melting medium plus 2.5 mL-1 of vegetable oil and an absolute control with PDA only. After two and four hours of incubation at 24ºC and constant light, germination of E. ampelina was assessed. The experimental design for in vitro experiments was completely randomized with four replicates and the experimental plot consisted of a Petri dish. In the field experiment, after pruning of the grapevine cv. Isabel, the levels of crude extract of garlic plus oil were weekly sprayed on the leaves of the plant, except on the absolute control (no treatment). From the first symptoms of anthracnose of grapevine, the severity that was expressed as area under the disease progress curve (AUDPC) was evaluated. The design was in randomized blocks with five replicates. The crude extract of garlic reduced the mycelial growth of the pathogen, especially when added to the culture medium prior to sterilization, when it expressed its maximal antifungal potential. When the vegetable oil was added to the extract levels, there was complete inhibition at the levels 25 or 30 mL L-1. This additive effect of these compounds was also noted for the germination test of E. ampelina. Under field conditions, the crude extract of garlic reduced the AUDPC by 83.59% at 25 mL L-1.

Persistence of Helicobacter pylori in the oral cavity after systemic eradication therapy.

AIM: The present study aimed to evaluate if the oral cavity of chronic periodontitis patients can harbor Helicobacter pylori after systemic eradication therapy. MATERIALS AND METHODS: Samples of 30 patients (15 with gingivitis and 15 with chronic periodontitis) positive for H. pylori in the stomach were evaluated. Samples were collected 3 months after triple systemic antibiotic therapy from saliva, microbiota from the dorsum of the tongue, supra- and sub-gingival plaque as well as gastric biopsies. DNA of each sample was extracted by the boiling method and used as a template in polymerase chain reaction with the primers JW22/23. RESULTS: Eighteen patients (60%) harboured H. pylori in their mouths. Five patients (16.6%) were positive in saliva, two (6.6%) on the dorsum of the tongue, nine (30%) in supra-gingival plaque, 14 (46.6%) in sub-gingival plaque and three (10%) in the stomach. There was no statistically significant difference between study groups. CONCLUSION: Eradication of H. pylori after therapy was more effective for the stomach than for the mouth (p<0.001). Mouths of patients with gingivitis or with chronic periodontitis, who are positive for H. pylori in their stomachs, may be considered as reservoirs of these bacteria.

Recorrência da infecção por H. pylori na Unifesp / Recurring H. pylori infection at Unifesp

Informações quanto à recorrência da infecção por H. Pylori (Hp) são restritas. A erradicação da bacteriaresulta na cura da doença ulcerosa péptica (DUP) na ausência de antiinflamatório. a DUP, todavia, recidiva com a reinfecção. Quanto maior a falha na erradicação maior a recorrencia, que envolve reinfecção e recrudescência. Objetivo: Avaliar o índice de recorrência do Hp ao longo do período pós-erradicação e confrontar os resultados do serviço com os observados em outros centros. Casuística e método: estudo retrospectivo longitudinal de pacientes com DUP infectados pelo Hp, atendidos de março de 1993 a agosto de 2003 no Ambulatório de Dispepsia da Disciplina e que receberam integralmente esquema para erradicação da bactéria. Foi feita reavaliação clínica, controle endoscópico com pesquisa de Hp, aos três , seis e 12 meses da erradicação e a partir de então, anualmente, por até oito anos. Para análise dos dados empregou-se o teste do qui-quadrado. Resultados: A 422 pacientes forneceram-se 609 tratamentos. Ao longo de oito anos, houve 633,4 pacientes-ano, o que denota o grande abandono de acompanhamento. A taxa de sucesso em três meses, seis meses e em um ano do tratamento foi de 70,3por cento,61,9por cento e de 54,2por cento, respectivamente. Considerando esquema com a associação do IBP, amoxilina e claritromicina (esquema mais eficaz dentre os utilizados) e somente a erradicação pela primeira vez, a taxa de sucesso foi de 85,3por cento, 81,0por cento, e de 74,3 em três meses, seis meses e em um ano após a erradicação, respectivamente. Conclusões:1) a taxa de sucesso de erradicação do Hp depende do intervalo de tempo em que se define o controle de cura, sendo o de três meses o mais adequado em relação ao de um mês; 2) há melhor resposta terapêutica na primeira tentativa de erradicação com esquema IBP, amoxicilina e claritromicina; 3) a positivação da pesquisa de Hp ocorre principalmente no primeiro ano após a erradicação; 4) quanto menor a falha terapêutica no primeiro ano, menores as taxas de recorrência ao longo dos anos; 5) de forma geral, a recorrência da infecção por Hp é de 9,6por cento ao ano (média ao longo de oito anos); considerando-se a associação IBP, amoxicilina e claritromicina, a média cai para 6,0por cento no mesmo período

Prevalence of Helicobacter pylori detected by polymerase chain reaction in the oral cavity of periodontitis patients.

Helicobacter pylori is an important gastrointestinal pathogen associated with gastritis, peptic ulcers, and an increased risk of gastric carcinoma. The oral cavity has been indicated as a possible H. pylori reservoir, and may therefore be involved in the reinfection of the stomach which sometimes follows treatment of H. pylori infection. The objective of the present study was to evaluate the prevalence of H. pylori as detected by polymerase chain reaction (PCR) in the oral cavity of periodontitis patients testing positive for this bacterium in the stomach. Thirty adult patients with alterations of the superior digestive tract, testing urease positive after endoscopy and biopsy, were selected. A full-mouth periodontal examination was performed in every patient and the subjects were allocated to two groups: gingivitis (15 patients) and chronic periodontitis (15 patients). Plaque and saliva samples collected from each patient were stored in 0.5 ml of TE buffer. DNA was extracted from the samples by the boiling method and was evaluated for the presence of H. pylori using the PCR method. JW 22/23 primers were used. The DNA of ATCC H. pylori 43629 (positive control) and water (negative control) were used for controlling the reactions. Of the 30 evaluated patients, 13 (43.3%) harbored H. pylori in the mouth. The bacterium was not found on the dorsum of the tongue of any patient, but was found in saliva in three patients (10%), in the supragingival plaque in six patients (20%), and in the subgingival plaque in eight patients (26.6%). The presence of H. pylori was similar in the gingivitis and chronic periodontitis groups. In conclusion, a high percentage of patients harbored H. pylori in their mouth. The bacterium was detected in saliva, supragingival and subgingival plaque, suggesting that these sites may be considered reservoirs for H. pylori in urease-positive patients.

Evolution of the nutritional support pattern in pediatric intensive care.

OBJECTIVES: To evaluate patterns of usage and monitoring of nutritional support in a Pediatric ICU of a teaching hospital and the role of an education program in nutritional support given throughout the resident physician training. DESIGN: In a historical cohort study, records from children who received nutritional support during the year 1992 were analyzed. Thereafter a continuing education program in Nutritional Support was conveyed to the residents. In a second phase of the study, the same parameters were reevaluated in children who received nutritional support throughout the year 1995. SETTING: Pediatric Intensive Care Unit of Department of Pediatrics, Escola Paulista de Medicina. PATIENTS: All the children who were given nutritional support during a period of five days or more. Based on this criteria 37 children were selected for the first phase of this study, and 35 for the second one. INTERVENTION: The education program included theoretical lectures about basic themes of nutritional support and journal article reading sessions. It was given to successive groups of residents on a weekly schedule. MEASUREMENTS: Daily records of fluid, protein, caloric and micronutrient supply, nutritional assessment and metabolic monitoring. RESULTS: In the first phase of the study, an exclusively parenteral route was utilized for 80.5%, and a digestive route 19.5% of the time period. Nutritional assessment was performed on 3 children; no patient had the nutritional goals set. The nitrogen to nonprotein calories ratio and the vitamin supply were inadequate, whilst the supply of trace elements was adequate except for zinc. Nutritional monitoring was performed on almost all patients but without uniformity. In the second phase, the exclusive parenteral route was used for 69.7% and the digestive route for 30.3% of the time period; no significant increase in the use of the digestive route was detected. The nonprotein calories to nitrogen ratio and micronutrient supply were adequate. The frequency of nutritional assessment increased, but deficiency in nutritional monitoring and infrequent enteral feeding were still detected. CONCLUSION: There were deficiencies in the implementation of nutritional support, which were partially corrected in the second phase of the study by the training of the residents. Reinforcement of the education program, which should be applied to the whole medical staff, and the organization of a multidisciplinary team in charge of coordinating the provision of nutritional support are suggested.

Suporte nutricional-metabólico em UTI pediátrica / Nutricional and metabolic support in the pediatric ICU

O suporte nutricional tem sido considerado um recurso terapêutico importante no tratamento de crianças hospitalizadas e gravemente doentes. Com o conhecimento do padrao de suporte nutricional utilizado pela unidade de internaçao hospitalar, pode-se detectar falhas e propor medidas para a sua otimizaçao. OBJETIVO. Avaliar o padrao terapêutico e de monitorizaçao do suporte metabólico em uma Unidade de Terapia Intensiva Pediátrica e propor medidas mínimas para melhorar a sua qualidade. PACIENTES E MÉTODO. Foram analisados os prontuários de 37 crianças admitidas na unidade durante um ano e que receberam suporte nutricional por um período igual ou superior a 15 dias. RESULTADOS. Em um período de 425 dias de suporte nutricional, a via parenteral exclusiva foi utilizada em 8O,5 por cento do tempo e a via digestiva (sonda nasoenteral ou oral), em 19,5 por cento do tempo. A avaliaçao nutricional prévia foi realizada em apenas três pacientes (8,1 por cento). Apenas 29,7 por cento das crianças tiveram suas necessidades energéticas satisfeitas, e este objetivo foi alcançado apenas nas que estavam recebendo nutriçao enteral. A relaçao nitrogênio/calorias nao-protéicas variou de 1:8O a 1:250. A oferta de oligoelementos por via parenteral foi adequada, exceçao feita ao zinco. A administraçao das vitaminas A, B2, E, C, pantenol e niacina esteve aquém da recomendada; biotina, vitamina B12, e folado nao foram administrados. A monitorizaçao nutricional incluiu o peso corpóreo e as dosagens de albumina sérica e de triglicérides, e foi realizada na maior parte dos pacientes, embora de modo eventual e nao uniforme. Conclusao. Houve falhas na implementaçao do suporte nutricional, a saber: inadequaçao da oferta protéica e de micronutrientes, monitoraçao nutricional e metabólica deficiente e uso pouco freqüente de nutriçao por via enteral. Sugere-se uma rotina mínima para a monitorizaçao de pacientes em nutriçao parenteral e enteral e a organizaçao de uma equipe multidisciplinar encarregada da coordenaçao do suporte nutricional.

[Nutritional and metabolic support in the pediatric ICU]. / Suporte nutricional-metabólico em UTI pediátrica.

UNLABELLED: Nutritional support has been considered an important part of the treatment of critically ill patients. The information about the current clinical pattern of nutrition support in hospitals may provide a basis for future modification and improvement of its prescription and use. OBJECTIVES: 1) To evaluate patterns of usage and monitoring of nutritional support in critically ill children; 2) To recommend policies aiming at the improvement of the nutritional support quality. PATIENTS AND METHOD: Records of 37 patients receiving nutritional support throughout one year were reviewed. RESULTS: From a total of 425 days of therapy, the single parenteral route was utilized in 80.50% the digestive route (tube feeding or oral route) in 19.5% of this time. A previous nutritional assessment was performed in 3 children; no patient had the nutrition goals set. The nitrogen to nonprotein calories ratio ranged between 1:80 and 1:250. Only 29.7% of the patients had their estimated caloric needs supplied and this goal was achieved only in those patients who were on enteral tube feeding. Patients did not achieved their goals for vitamins. The supply of oligoelements was adequate except zinc. Nutritional monitoring parameters including weight, serum albumin and serum triglycerides were performed in almost all the patients but without uniformity. CONCLUSION: There was a lack in the implementation of nutritional support. Inadequacy of protein and micronutrients supply, irregular nutritional monitoring and infrequent enteral feeding were detected. A minimal standard for nutritional and metabolic monitoring and the organization of a multidisciplinary team in charge of coordinating the providing of nutritional support are suggested.