Octyl gallate, a food additive with potential beneficial properties to treat Helicobacter pylori infection.

Helicobacter pylori infection is marked by intense production of reactive oxygen species (ROS) through the activation of neutrophils that are constantly attracted to the infected gastric mucosa. Here, gallic acid and its alkyl esters were evaluated as compounds able to act as antimicrobial agents and inhibitors of ROS released by H. pylori-activated neutrophils simultaneously. We found that the higher hydrophobicity caused by esterification of gallic acid led to a significant increase in its ability as a cytotoxic agent against H. pylori, a scavenger of ROS and an inhibitor of NADPH oxidase in neutrophils. Octyl gallate, a widely used food additive, showed the highest antimicrobial activity against H. pylori, with a minimum inhibitory concentration (MIC) value of 125 µg mL-1, whereas gallic acid had a MIC value higher than 1000 µg mL-1. The production of superoxide anion radicals was almost 100% abolished by the addition of 10 µM (2.82 µg mL-1) octyl gallate, whereas gallic acid inhibited around 20%. A similar tendency was also found when measuring the production of hypochlorous acid. The protective effect of the esters was cytochemically confirmed. In conclusion, this study showed that hydrophobicity is a crucial factor to obtain a significant anti-ROS and anti-H. pylori activity. Finally, it highlights octyl gallate, a food additive widely used in the food industry, as a promising molecule in the treatment of H. pylori infection.

Alkyl caffeates as anti-Helicobacter pylori and scavenger of oxidants produced by neutrophils.

Helicobacter pylori pathogenic action involves the colonization of the gastrointestinal tract and a large production of reactive oxygen species (ROS) by the neutrophils attracted to the site of infection. The aim of this study was to evaluate caffeic acid and its alkyl esters as inhibitors of the release of ROS by Helicobacter pylori activated neutrophils and their bactericidal effect. The increased hydrophobicity caused by esterification had direct consequence in their efficiency as bactericidal agents against H. pylori and inhibitors of the production of ROS by neutrophils. The minimum inhibitory concentration (MIC) decreased from higher than 1000 µg/mL (caffeic acid) to 250 µg/mL to butyl and heptyl caffeate. The release of total ROS, superoxide anion and hypochlorous acid by activated neutrophils was also significantly decreased and the esters were more efficient than the acid precursor. In conclusion, the alkyl esters of caffeic acid have two properties that are complementary for the treatment of H. pylori infections: bactericidal activity and inhibitory effect upon generation of ROS by neutrophils. Hence, we propose that these easily synthesized and non-expensive substances should be applied to in vivo experimental models of H. pylori induced gastric infections.

Comparison of Brazilian Plants Used to Treat Gastritis on the Oxidative Burst of Helicobacter pylori-Stimulated Neutrophil.

Ten Brazilian medicinal plants used to treat gastritis and ulcers were carefully selected on the basis of ethnopharmacological importance and antiulcerogenic activity previously described. The antioxidant activity of the methanolic extracts was determined in analysis conditions that simulate a real biological activity on inhibition of the oxidative burst induced in neutrophils using Helicobacter pylori as activator, by a luminol-amplified chemiluminescence assay. The extracts, at low concentration (5 µ g/mL), exhibited a large variation in inhibitory effects of H. pylori-induced oxidative burst ranging from 48% inhibition to inactive, but all extracts, excluding Byrsonima intermedia, had inhibitory activity over 80% at the concentration of 100 µ g/mL. The total suppressive antioxidant capacity measured as the effective concentration, which represents the extract concentration producing 50% inhibition of the chemiluminescence induced by H. pylori, varies from 27.2 to 56.8 µ g/mL and was in the following order: Qualea parviflora > Qualea multiflora > Alchornea triplinervia > Qualea grandiflora > Anacardium humile > Davilla elliptica > Mouriri pusa > Byrsonima basiloba > Alchornea glandulosa > Byrsonima intermedia. The main groups of compounds in tested extracts are presented. Differences in the phytochemical profile, quantitatively and qualitatively, of these plants can explain and justify their protective effect on the gastric mucosa caused by the neutrophil-generated ROS that occurs when H. pylori displays its evasion mechanisms.

Inhibition of nitric oxide and tumour necrosis factor-α production in peritoneal macrophages by Aspergillus nidulans melanin.

The naturally occurring pigment, melanin is found in organisms of all phylogenetic kingdoms, including fungi, and exhibits a wide range of biological activities. Our objective was to investigate the effects of melanin extracted from the fungus Aspergillus nidulans on the production of the pro-inflammatory mediators nitric oxide (NO) and tumour necrosis factor-α (TNF-α) in peritoneal macrophages and on the viability of McCoy mouse fibroblasts. The results showed that A. nidulans melanin did not stimulate NO production in macrophages, but it inhibited the NO production in lipopolysaccharide (LPS)-stimulated macrophages by approximately 82%. Similarly, A. nidulans melanin inhibited LPS-stimulated TNF-α production by 52% and showed a slight stimulatory effect on TNF-α production in macrophages. In addition, the toxicity of A. nidulans melanin to McCoy cells was much lesser (IC50=373.5±2.4 µg/mL) than that of known agents such as cisplatin (IC50=41.2 µg/mL). The viability of peritoneal macrophages was greater than 90% at the highest melanin concentration tested (100 µg/mL). Thus, the combination of low cytotoxicity and marked inhibition of TNF-α and NO production suggests that A. nidulans melanin has potential as an anti-inflammatory agent and may be used in the future for development of new drugs with therapeutic utility.

Anti-Helicobacter pylori activity and oxidative burst inhibition by the naphthoquinone 5-methoxy-3,4-dehydroxanthomegnin from Paepalanthus latipes

Helicobacter pylori is a bacterium recognized as the major cause of chronic gastritis and peptic ulcers. Infection by H. pylori induces inflammatory responses and pathological changes in the gastric microenvironment. The host Keywords: immune cells (especially neutrophils) release inflammatory mediators and large 5-methoxy-3,4-dehydroxanthomegnin amounts of reactive oxygen species (ROS), which are associated with an increased Helicobacter pyloririsk of developing gastric cancer. In this study, we evaluated the anti-H. pylori and oxidative burst antioxidantactivitiesofa1,4-naphthoquinone-5-methoxy-3,4-dehydroxanthomegnin. Paepalanthus latipes The antimicrobial activity was assessed using a spectrophotometric microdilution technique, and antioxidant activity was assessed by noting the effect of 5-methoxy3,4-dehydroxanthomegnin on the neutrophil oxidative burst using luminol-and lucigenin-amplified chemiluminescence. The results showed that 5-methoxy-3,4dehydroxanthomegnin is a potent anti-H. pylori compound (MIC 64 µg/mL and MBC 128 µg/mL) and a strong antioxidant. 5-Methoxy-3,4-dehydroxanthomegnin decreased luminol- and lucigenin-amplified chemiluminescence, with ED50 values of 1.58±0.09 µg/mL and 5.4±0.15 µg/mL, respectively, reflecting an inhibitory effect on the oxidative burst. These results indicate that 5-methoxy-3,4-dehydroxanthomegnin is a promising compound for the prevention and treatment of diseases caused by H. pylori infection, such as gastritis, peptic ulceration, and gastric cancer, because reactive oxygen intermediates are involved in the pathogenesis of gastric mucosal injury induced by H. pylori infections.

Antitumor and immunomodulatory effects of the naphthoquinone 5-methoxy-3,4-dehydroxanthomegnin

Large number of quinones has been associated with antitumor, antibacterial, antimalarial and antifungal activities. In this work we describe the effect of the naphthoquinone, 5-methoxy-3,4-dehydroxanthomegnin, on murine tumor cells (LP07 and LM2) and its immunomodulatory effect on nitric oxide (NO) production on LPS-stimulated macrophages. The results have shown that 5-methoxy-3,4-dehydroxanthomegnin was a significant inhibitor of LPS-stimulated NO generation from macrophage (inhibition percentage ranged from 97.4 to 98.9 percent) and a strong cytotoxic agent against both tumor cells LP07 and LM2 (CI50 6.2±0.36 µM and 74.6±1.9 µM, respectively). These results indicate that the 5-methoxy-3,4-dehydroxanthomegnin may show promising activity in the treatment of murine breast and lung cancer by immunomodulatory and antiproliferative activities.

The triad Indole-3-Acetic Acid Ethyl Ester/Esterase/Horseradish Peroxidase as a new cytotoxic Prodrug/Enzyme combination

The antibody-directed enzyme prodrug therapy (ADEPT) is a means of restricting the action of toxic drugs to the tumor site. The enzyme/prodrug pair horseradish peroxidase (HRP)/indole-3-acetic acid (IAA) has been studied as a combination with potential application in ADEPT strategies. In this combination, the non-toxic plant hormone IAA is activated to cytotoxic species by the catalytic action of HRP. Objective: We studied the use of the ethyl ester of IAA as a new prodrug that could be activated by two enzymes, HRP and esterase. Methods: The oxidation of IAA and its ethyl ester, catalyzed by HRP, was monitored by the consumption of dioxygen and liquid chromatography. The cytotoxicity of IAA and its ethyl ester in combination with HRP and esterase was assessed using the lineage McCoy cells through the trypan blue and neutral red assays. Results: We found that HRP was not able to catalyze the oxidation of IAA-ethyl ester in the absence of an additional esterase. Hence, the potential cytotoxicity of the IAA-ethyl ester could be controlled by sequential treatment with esterase, to liberate the carboxyl group, and HRP, for oxidation and generation of cytotoxic species. We present evidence for the potential application of the combination IAA-ethyl ester/esterase/horseradish peroxidase as a new ADEPT, GDEPT or related strategy. Conclusions: We suggest that this technique could provide more selectivity in the generation of cytotoxic drugs at tumor sites.

Indoor Air quality related to occupancy at an air-conditioned public building

To characterize the influence of occupancy on the indoor air quality, a public office building with air-conditioning system was selected for this study. The indoor parameters included total bacteria count, total fungal count, temperature, relative humidity, and carbon dioxide concentration. The number of occupants, which varied throughout the day, was recorded in each sample. The samples were taken before the beginning of the working day and during 3 h, at an interval of 30 min between each sampling, and continued for five working days during a week. Correlation analysis demonstrated that occupancy rates were positively correlated with airborne bacteria, CO2, and temperature. No significant association between the number of occupants and fungus was observed. The results of this study provided information on the variability of indoor air parameters during the time-varying occupancy over the course of the day in at air-conditioned buildings where occupancy was quite dynamic.

Com o objetivo de caracterizar a influência da ocupação na qualidade do ar interior, um edifício público com sistema de ar condicionado foi selecionado. As variáveis ambientais consideradas incluíram contagem total de bactérias e fungos, temperatura, umidade relativa e concentração de dióxido de carbono. O número de ocupantes, que variou durante todo o dia, foi estimado em cada amostragem. As amostras foram coletadas antes do início do expediente de trabalho e durante 3 horas, em intervalos de 30 minutos, por 5 dias úteis consecutivos. A análise de correlação demonstrou que a taxa de ocupação foi correlacionada positivamente com a concentração de bactérias, dióxido de carbono e temperatura. Nenhuma associação significativa foi observada entre o número de ocupantes e concentração de fungos. Os resultados deste estudo fornecem informações quanto à variabilidade nos parâmetros do ar interior no decorrer do dia em um edifício onde a ocupação é dinâmica.

Effect of subinhibitory concentration of chlorhexidine on Streptococcus agalactiae virulence factor expression.

The aim of the present study was to investigate the effect of chlorhexidine at subinhibitory concentration (50% minimal inhibitory concentration (MIC)) on the growth, cytolysin expression and phagocytosis of Streptococcus agalactiae ATCC 13813. Bacterial growth with and without chlorhexidine treatment was monitored by turbidity measurements, and exocytolysins were estimated by neutral red uptake assay by the McCoy cell line. The phagocytic process was evaluated using luminol-enhanced chemiluminescence to follow the respiratory burst of polymorphonuclear neutrophils exposed to bacteria. Chlorhexidine-treated culture did not exhibit a detectable decrease in cell growth, and no statistically significant reduction in the respiratory burst of polymorphonuclear neutrophils was observed. However, growth in the presence of chlorhexidine resulted in a significant reduction of S. agalactiae exocytolysins. Although 50% MIC of chlorhexidine did not interfere with S. agalactiae growth and phagocytosis, the knowledge that this concentration was still able to alter some bacterial virulence parameters may be useful in its therapeutic applications.

Mutagenic and cytotoxic effect of planifolin: a naphthopyranone dimer isolated from Paepalanthus planifolius.

A naphthopyranone dimer, named planifolin, was isolated from a methylene chloride extract of the capitula of Paepalanthus planifolius Koern. The molecule (C(31)H(26)O(10)) appeared to be made up of two monomeric portions, semi-vioxanthin and paepalantine (an isocoumarin), linked by an ether bond, and it may possess several kinds of biological activity that can be related to its polyphenolic structure. Short-term tests that detect genetic damage can afford the information needed to evaluate carcinogenic risks of chemicals to humans. The Ames test, recommended for testing the mutagenicity of chemical compounds with potential pharmacological application, was used in the present study. The mutagenic activity was evaluated in Salmonella typhimurium strains TA100, TA98, TA102 and TA97a and the cytotoxic effect in McCoy cells. The in vitro cytotoxicity of planifolin to McCoy cells, tested in microculture with neutral red, showed a significant cytotoxic index (CI(50)) of 12.83 microg/mL. Planifolin showed mutagenic activity for TA100, TA98 and TA97a. The results indicate that this new naphthopyranone dimer causes mutations by substitution and by addition and deletion of bases in the sequence of DNA. Moreover, its mutagenic potential was increased by metabolic activation.

A new cytotoxic naphthoquinone from Paepalanthus latipes.

Quinones constitute an important class of naturally occurring compounds. They are found in plants, fungi and bacteria. Large number of quinones has been associated with antitumor, antibacterial, antimalarial and antifungal activities. In this work we describe the isolation, structure determination and the cytotoxic index of a new 1,4-naphthoquinone isolated from the capitula of Paepalanthus latipes.

Effect of the isocoumarin paepalantine on the luminol and lucigenin amplified chemiluminescence of rat neutrophils.

Paepalantine (9,10-dihydroxy-5,7-dimethoxy-1H-naphto(2,3c)pyran-1-one), a natural isocoumarin isolated from the capitula of Paepalanthus bromelioides (Eriocaulaceae), was assessed for its effect on the respiratory burst (zymosan-stimulated luminol-enhanced chemiluminescence and PMA-stimulated lucigenin-enhanced chemiluminescence) of polymorphonuclear neutrophils in vitro. Special attention was devoted to establishing the IC(50) for neutrophils. Paepalantine was able to decrease luminol and lucigenin chemiluminescence, reflecting an inhibitory effect on the respiratory burst, with an ED(50) of 0.44+/-0.05 and 0.84+/-0.15 microg/ml, respectively. A cell-free system was performed with paepalantine on myeloperoxidase/H(2)O(2) and myeloperoxidase/H(2)O(2)/Cl(-) systems. Paepalantine inhibited luminol oxidation in both systems. This inhibition was related to the interaction of paepalantine-myeloperoxidase and its scavenger effect on HOCl.

Avaliação citológica da mucosa nasal de portadores de staphyloccus aureus / Staphylococcus aureus' carriers nasal mucosal cytologic evaluation

Voluntários (32) que atuam profissionalmente na recepção de Postos de Saúde da cidade de Araraquara participaram desse estudo. Para a coleta do lavado nasal, 5 mL de solução salina estéril foram instilados em cada narina e recolhidos em recipiente estéril. O fluído recuperado foi centrifugado e o sedimento ressuspendido em solução fisiológica (1,5 mL). Uma alíquota foi cultivada em agar sangue de carneiro (5 porcento) objetivando o isolamento de S.aureus. As cepas isoladas foram submetidas ao teste de susceptibilidade a alguns antimicrobianos. A contagem total de células presente foi realizada em câmara de Neubauer e a citologia diferencial através de esfregaços corados pelo método de Grunwald-Giemsa. Os resultados demonstraram que portadores do microrganismo apresentam neutrófilos polimorfonucleares em quantidade significativamente aumentada em relação aos não portadores, indicando resposta celular aguda. S. aureus é o agente mais comum de infecções piogênicas e apresentam vários componentes de superfície que contribuem para a quimiotaxia de leucócitos, justificando assim o aumento desse tipo celular.

Genotoxicity of Brosimum gaudichaudii measured by the Salmonella/microsome assay and chromosomal aberrations in CHO cells.

The root bark of Brosimum gaudichaudii Trécul (Moraceae) is popularly used for treatment of vitiligo. In the present study the mutagenic activity of the aqueous and methanolic extract as well as of the n-butanolic fraction of this medicinal plant were evaluated using Salmonella typhimurium assays, TA100, TA98, TA102 and TA97a strains, while the clastogenic effect in Chinese hamster ovary (CHO) cells in the G(1)/S, S and G(2)/S phases of the cell cycle. The results showed mutagenic activity of the aqueous extract against TA102 in the presence of S9, and of methanolic extract, with and without metabolic activation. TA100 mutagenicity was only observed for the methanolic extract in the absence of S9. The n-butanolic fraction did not present mutagenic activity. In CHO cells only the methanolic extract induced a significant increase of chromosomal aberrations in the G(1)/S and S phases, whereas a decrease in the mitotic index was observed in the G(1)/S and G(2)/S phases. No clastogenicity was observed for the aqueous extract. The furocoumarins (psoralen and bergapten) presented in the extracts might contribute to the mutagenicity. The lower activity of the aqueous extract was probably due to the presence of smaller amount of furocoumarins compared to the methanolic extract.

Bioaerossóis em ambientes do prédio tradicional da Faculdade de Ciências Farmacêuticas - UNESP / Microbial bioaerosols in environments of the Traditional Building at the School of Pharmaceutical Sciences

Este trabalho objetivou conhecer a concentraçäo microbiana e a influência de alguns fatores ambientais na dispersäo de bioaerossóis em cômodos específicos, de uso restrito e público, do Prédio Tradicional da Faculdade de Ciências Farmacêuticas, onde situa o Departamento de Análises Clínicas, Farmácia Escola e Unidade Auxiliar. As amostragens foram realizadas utilizando-se o MAS-100, durante 2 períodos do ano, em 15 cômodos, sendo 5 laboratórios didáticos, 5 laboratórios de rotina e 5 salas de atendimento ao público. Os ambientes foram analisados em relaçäo ao tipo de climatizaçäo (natural ou artificial) e ao número de ocupantes. O tipo de climatizaçäo demonstrou näo influenciar no número de unidades formadoras de colônia por metro cúbico de ar dos diferentes ambientes. Diferenças significativas foram observadas em ambientes onde circulavam menos de 9 e 9 ou mais pessoas, independentemente do tipo de atividade desenvolvida. O fato de os próprios ocupantes serem as fontes mais prováveis de bioaerossóis em ambientes interiores estimula a instalaçäo de equipamentos eficientes que promovam a filtraçäo e renovaçäo do ar, diluindo partículas originadas dentro deles, principalmente naqueles onde há maior fluxo de indivíduos.

Citotoxicidade in vitro de soluçöes de digluconato de clorexidina preparadas em farmácias de manipulaçäo / In vitro cytotoxicity of mouthrinses of chlorhexidine digluconate prepared in manipulation pharmacies

O digluconato de clorexidina, agente antimicrobiano frequentemente utilizado na forma de soluçäo de bochecho a 0,12 por cento, tem demostrado ser efetivo na reduçäo da placa subgengival e gengivites. Estudos clínicos, contudo, relatam que soluçöes de clorexidina podem retardar o processo de cicatrizaçäo bem como causar lesöes descamativas. O presente trabalho avaliou in vitro o efeito citotóxico de soluçöes de bochecho, contendo, segundo os fabricantes, 0,12 por cento de digluconato de clorexidina, manipuladas e comercializadas na cidade de Araraquara, sobre células McCoy, dando ênfase ao agente responsável pela toxicidade. Os volumes citotóxicos (VC50) foram determinados pela técnica do MTT-tetrazólio.

Microbiota vaginal durante o ciclo menstrual / Vaginal microbiota during the menstrual cycle

Com o objetivo de detectar variaçöes da microbiota vaginal, pH e número de leucócitos polimorfonucleares nas diferentes fases do ciclo menstrual, dez mulheres assintomáticas foram amostradas por três semanas consecutivas e avaliadas hormonalmente através de estendidos citológicos corados pelo método de Papanicolaou. Lactobacilos anaeróbios facultativos predominaram nas amostras colhidas das fases pós-menstrual, ovulatória e pré-menstrual em 90 por cento das mulheres amostradas. Näo houve alteraçöes significativas da microbiota e pH vaginal. O número médio de leucócitos foi maior nos espécimes colhidos na fase pré-menstrual. Flutuaçöes hormonais, durante o ciclo menstrual, näo alteram significativamente a microbiota vaginal; entretanto, a presença de um número apreciável de leucócitos näo deve ser utilizada como critério para a triagem de pacientes portadoras de vaginite em espécimens colhidos na fase pós-menstrual.

Chlamydia trachomatis, ureaplasma urealyticum e outros patógenos uretrais em homens sexualmente ativos / Chlamydia trachomatis, ureaplasma urealyticum and others urethral pathogens in sexual active men

Sessenta e nove homens sexualmente ativos foram examinados quanto a presença de Chlamydiz trachomatis, Ureaplasma urealyticum e outros patógenos uretrais. Foram encontrados os seguintes microorganismos: C. trachomatis (24,6 por cento), U. urelyticum (19,1 por cento), Streptococcus agalactiae (15,9 por cento), Gardnerella vaginallis (1,4 por cento) e Candida sp (1,4 por cento). Outro objetivo do presente estudo foi relacionar a presença desses microorganismos aos sintomas referidos pelo paciente. C. Trachomatis e U. urealyticum foram isolados com maior freqüência em pacientes sintomáticos, S. agalactiae foi isolado, predominantemente, de pacientes assitomáticos. Semelhantemente aos sintomas subjetivos, os esfregaços corados pelo método Gram possuem valor limitado no diagnóstico das uretrites. Este estudo vem demonstrar a importância da identificação completa dos microorganismos isolados em material uretral para o controle das doenças sexualmente transmissíveis.

Adesäo de Mycobacterium tuberculosis, M. avium e de M. fortuitum em células McCoy / Adherence capacity of M. tuberculosis, M. avium and M. fortuitum on McCoy cells line

A capacidade de adesäo do M. tuberculosis, M. avium e M. fortuitum às células da linhagem McCoy foi analisada, em intervalos de tempo predeterminados. Foram estabalecidas para o ensaio concentraçäo de 10(5) células/ml de meio de Eagle, suspensäo inoculante das micobactérias na fase logarítima e incubaçäo sob agitaçäo de 60 rpm para se evitar adesäo inespecífica. Verificou-se adesäo mais rápida e eficiente do M. fortuitum às células McCoy em relaçäo às outras duas micobactérias

Comparaçäo do Papanicolau e cultura em células para diagnóstico de infecçöes cervicais causadas por Chlamydia trachomatis / Comparison of Papanicolaou smear and cell culture in the diagnosis of Chlamydia trachomatis cervical infections

Neste estudo, comparamos a técnica do Papanicolaou e cultura em células para a detecçäo de infecçöes cervicais causadas por Chlamydia trachomatis. A taxa de isolamento foi de 15,7 porcento. Utilizando como critério a presença de células metaplásicas contendo corpúsculos de inclusäo intracitoplasmáticos, 3 das 11 pacientes (27,3 porcento) com cultura positiva tiveram os esfregaços corados pelo Papanicolaou interpretados como sugestivos de infecçäo clamidiana e 10 das 59 mulheres (16,9 porcento) com cultura negativa apresentaram no Papanicolaou critérios morfológicos satisfatórios para o diagnóstico da infecçäo (sensibilidade de 27,3 porcento e especificidade de 83,1 porcento). Esses dados confirmam os obtidos por outros autores que a técnica de Papanicolaou em esfregaços cervicais é de baixa sensibilidade para o diagnóstico ou triagem de infecçöes clamidianas e näo deve ser utilizada com essa finalidade