Defensin c-thionin from Capsicum chinense improves butyrate cytotoxicity on human colon adenocarcinoma cell line Caco-2

BACKGROUND: Butyrate is a histone deacetylase inhibitor that induces apoptosis and inhibits cell proliferation of colorectal cancer cells. To improve its anticancer activity, butyrate has been evaluated mixed with drugs and different molecules. Plant antimicrobial peptides are attractive anticancer alternative molecules because they show selective cytotoxic activity against different cancer cell lines. In this work, we explore if the plant defensin c-thionin (Capsicum chinense) can improve butyrate activity on Caco-2 cell line and we also determined the mechanism of death activated. RESULTS: The combined treatment of c-thionin (3.5 mM) and butyrate (50 mM) showed higher cytotoxicity on Caco-2 cells with respect to single treatments. Also, the combined treatment reduced cell proliferation and exhibited a higher rate of apoptosis than single treatments. Combined treatment induced caspases 8 and 9 activation to an extent comparable with that of butyrate while c-thionin did not activate caspases. Additionally, reactive oxygen species generation preceded the onset of apoptosis, and superoxide anion production was higher in cells treated with the combined treatment. CONCLUSIONS: The c-thionin from Habanero chili pepper improved the butyrate cytotoxicity on Caco-2 cells. This effect occurred through apoptosis induction associated with reactive oxygen species production. Therefore, the combination of butyrate with cytotoxic antimicrobial peptides could be an attractive strategy for cancer therapy.

Sodium butyrate in post-hatch feed for broiler chicks / [Uso de butirato de sódio em rações pós-eclosão para pintos de corte]

This study was designed to determine adequate levels for sodium butyrate inclusion in pre-hatching and pre-starter feed in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, each comprising five replicates of ten birds each. Five dietetic levels of sodium butyrate (control, 0.050, 0.075, 0.100 and 0.200%) were used in the pre-starter feed offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated after 24 hours of feed access and at 7 days of age. A metabolic trial was performed when the chicks were seven to ten days of age. Supplementation of 0.1% sodium butyrate increased the development of broiler chicks' intestinal villi at seven days of age but was not able to improve performance in the pre-starter phase. Supplementation with sodium butyrate in a diet offered in the transportation box does not improve broiler performance in the pre-starter phase. Supplementation of up to 0.16% is recommended, in order to improve the metabolizability of the ether extract for broilers at seven days of age.(AU)

Objetivou-se, com este estudo, encontrar níveis adequados de inclusão de butirato de sódio em ração pós-eclosão e pré-inicial, buscando minimizar os efeitos negativos do jejum de pintos de corte. Os animais foram distribuídos em delineamento inteiramente ao acaso, com cinco tratamentos e cinco repetições de 10 aves cada. Foram utilizadas cinco suplementações de butirato de sódio (controle; 0,05; 0,075; 0,100 e 0,200%) na ração pré-inicial, fornecidas aos pintos na caixa de transporte e durante a fase pré-inicial. Após 24 horas de acesso ao alimento e aos sete dias de idade, foram avaliados desempenho, retração do saco vitelino, concentração de glicose plasmática, peso e histomorfometria do intestino delgado. De sete a 10 dias de idade, realizou-se ensaio de metabolizabilidade. A suplementação de 0,10% de butirato de sódio aumentou o desenvolvimento das vilosidades intestinais dos pintos de corte aos sete dias de idade, mas não foi capaz de melhorar o desempenho na fase pré-inicial. A suplementação com butirato de sódio em dieta oferecida na caixa de transporte não melhora o desempenho dos frangos na fase pré-inicial. Recomenda-se suplementação de até 0,16%, a fim de melhorar a metabolizabilidade do extrato etéreo para frangos de corte aos sete dias de idade.(AU)

Enzymatic synthesis optimization of isoamyl butyrate from fusel oil

Many food, cosmetic and pharmaceutical industries have increased their interest in short-chain esters due to their flavor properties. From the industrial standpoint, enzyme reactions are the most economical strategy to reach green products with neither toxicity nor damage to human health. Isoamyl butyrate (pear flavor) was synthesized by isoamyl alcohol (a byproduct of alcohol production) and butyric acid with the use of the immobilized lipase Lipozyme TL IM and hexane as solvents. Reaction variables (temperature, butyric acid concentration, isoamyl alcohol:butyric acid molar ratio and enzyme concentration) were investigated in ester conversion (%), concentration (mol L-1) and productivity (mmol ester g-1 mixture . h), by applying a sequential strategy of the Fractional Factorial Design (FFD) and the Central Composite Rotatable Design (CCRD). High isoamyl butyrate conversion of 95.8% was achieved at 24 hours. At 3 hours, the highest isoamyl butyrate concentration (1.64 mol L-1) and productivity (0.19 mmol ester g-1 mixture . h) were obtained under different reaction conditions. Due to high specificity and selectivity of lipases, process parameters of this study and their interaction with the Lipozyme TL IM are fundamental to understand and optimize the system so as to achieve maximum yield to scale up. Results show that fusel oil may be recycled by the green chemistry process proposed by this study.

Butirato de sodio como agente coadyuvante de la acción radioprotectora de la amifostina / Sodium butyrate as a coadyuvant agent of the radioprotecting action of amifostine / Butirato de sódio como agente coadjuvante da ação radioprotetora da amifostina

Entre los escasos radioprotectores en uso, la amifostina resulta eficaz para reducir la toxicidad aguda inducida por la radiación ionizante. Sin embargo, presenta efectos tóxicos importantes que impiden su uso repetido o en dosis altas. Es necesario entonces desarrollar radioprotectores menos tóxicos, por sí mismos o como coadyuvantes de la amifostina en dosis bajas. Se expusieron ratas Sprague-Dawley a una dosis de rayos X de 6 Gy (cuerpo entero). Se ensayó el butirato de sodio como mitigante luego de una dosis baja de amifostina previa a la irradiación. A distintos tiempos después de la irradiación se realizó el recuento de eritrocitos, leucocitos y la fórmula leucocitaria. Los efectos genotóxicos se evaluaron en leucocitos de sangre mediante el ensayo Cometa. Se realizaron también estudios de supervivencia a 60 días y la evaluación histológica del duodeno e intestino grueso. El efecto del tratamiento resultó moderadamente protector respecto de la recuperación de los valores normales de eritrocitos, leucocitos y la fórmula leucocitaria en los animales sobrevivientes en ambos sexos, así como de los epitelios intestinales y el ADN de los leucocitos. También aumentó significativamente la sobrevida a 60 días. La radioprotección con amifostina en una dosis baja seguida de una mitigación con butirato fue claramente significativa.

Among the few radioprotectors in use, amifostine is effective in reducing the acute toxicity induced by ionizing radiation. However, it has important toxic effects that prevent its repeated use or in high doses. It is necessary then to develop less toxic radioprotectors, by themselves or as adjuvants of amifostine in low doses. Sprague-Dawley rats were exposed to an X-ray dose of 6 Gy (whole body). Sodium butyrate was tested as a mitigant after a low dose of amifostine prior to irradiation. At different times after the irradiation, the erythrocytes, leukocytes and the leukocyte formula were counted. Genotoxic effects were evaluated in blood leukocytes by the Comet assay. Sixty-day survival studies and histological evaluation of the duodenum and large intestine were also performed. The effect of the treatment was moderately protective with respect to the recovery of the normal values of erythrocytes, leukocytes and the leukocyte formula in the surviving animals in both sexes as well as for the intestinal epithelia and leukocytes DNA. It also significantly increased the 60-day survival. The radioprotection with amifostine in a low dose followed by mitigation with butyrate was clearly significant.

Entre os poucos radioprotetores em uso, a amifostina é eficaz na redução da toxicidade aguda induzida pela radiação ionizante. No entanto, tem importantes efeitos tóxicos que impedem seu uso repetido ou em altas doses. É necessário, então, desenvolver radioprotetores menos tóxicos, isoladamente ou como coadjuvantes da amifostina em baixas doses. Ratos Sprague-Dawley foram expostos a uma dose de raios X de 6 Gy (corpo inteiro). O butirato de sódio foi testado como mitigante após uma dose baixa de amifostina antes da irradiação. Em diferentes momentos após a irradiação, os eritrócitos, leucócitos e a fórmula de leucócitos foram contados. Os efeitos genotóxicos foram avaliados em leucócitos de sangue pelo ensaio Cometa. Estudos de sobrevida de 60 dias e avaliação histológica do duodeno e do intestino grosso também foram realizados. O efeito do tratamento resultou moderadamente protetor em relação à recuperação de valores normais de eritrócitos, leucócitos e fórmula leucocitária nos animais sobreviventes em ambos os sexos, bem como protegeu epitélios intestinais e o DNA dos leucócitos. Também aumentou significativamente a sobrevida para 60 dias. A radioproteção com amifostina em baixa dose seguida de uma mitigação com butirato foi claramente significativa.

Dermatite de contato por corticoide em pacientes com dermatite atópica / Corticosteroid contact dermatitis in patients with atopic dermatitis

Objetivo: Avaliar a sensibilização a corticoides tópicos e substâncias do teste de contato padrão e cosméticos em pacientes com dermatite atópica (DA) no Serviço de Alergia e Imunologia do HSPE-SP. Método: Estudo retrospectivo com análise de prontuário de pacientes com DA, classificados de acordo com o SCORing Atopic Dermatitis (SCORAD) em leve, moderada e grave, que foram atendidos no ambulatório de Alergia e Imunologia do HSPE-SP, submetidos a teste de contato com baterias padrão, cosméticos, corticoides, incluindo furoato de mometasona. Resultados: Após análise estatística dos dados de 51 pacientes portadores de DA, foi identificada maior prevalência no gênero feminino (73%). Pacientes com DA moderada/grave apresentaram maior positividade (62%) para pelo menos uma substância. Foram mais propensos a positivar para teste de contato, pacientes maiores de 18 anos. As substâncias que foram mais positivas na bateria padrão foram: sulfato de níquel (33%), neomicina (10%), e bicromato de potássio e cloreto de cobalto (8% cada). O sulfato de níquel foi mais positivo no gênero feminino. Três (5,9%) pacientes apresentaram positividade para teste de contato com bateria de corticoides, sendo positivas substâncias betametasona 1%, budesonida 0,01% e butirato de hidrocortisona 1%, e todos eram portadores de DA leve. Foi identificada relação entre positividade para bateria de corticoides e sulfato de níquel. Conclusão: Os testes de contato foram mais positivos em adultos. Houve maior sensibilidade para o sulfato de níquel no gênero feminino. Sensibilidade importante à neomicina na DA moderada/grave. Pacientes com alergia de contato por corticoides podem apresentar alergia a sulfato de níquel. Esse trabalho chama atenção para a porcentagem importante de pacientes com DA acometidos por alergia de contato por corticoides, sendo esse tipo alergia um problema emergente e que tem sido cada vez mais relatado na última década; porém, ainda são escassos os estudos envolvendo esse assunto.

Objective: To evaluate sensitization to topical corticosteroids and standard patch testing substances in patients with atopic dermatitis (AD) at the HSPE-SP Allergy and Immunology Department. Methods: This retrospective study assessed medical records of patients with AD, whose severity was classified as mild, moderate or severe according to SCORing Atopic Dermatitis (SCORAD). They were seen at the HSPE-SP Allergy and Immunology outpatient clinic and underwent a patch test with the following series: standard, cosmetics, and corticosteroids, including mometasone furoate. Results: After statistical analysis of data from 51 patients with AD, a higher prevalence was identified in female participants (73%). Patients with moderate-to-severe AD had more positive results (62%) for at least one substance. Patients older than 18 years were more likely to be positive in the patch test. The substances that were most frequently positive in the standard series were nickel sulfate (33%), neomycin (10%) and potassium dichromate and cobalt chloride (8% each). Positive nickel sulfate was more common in female participants. Three (5.9%) patients were positive for corticosteroids (1% betamethasone, 0.01% budesonide and 1% hydrocortisone butyrate) and all had mild AD. A relationship between positivity for corticosteroid series and nickel sulfate was identified. Conclusion: Patch tests were more frequently positive in adults. There was higher sensitivity to nickel sulfate in female patients. There was important sensitivity to neomycin in patients with moderate-to-severe AD. Patients with corticosteroid contact allergy may present with allergy to nickel sulfate. This paper draws attention to the important percentage of patients with AD affected by corticosteroid contact allergy, which has become an emerging problem that has been increasingly reported in the past decade. However, there are still few studies addressing this topic.

Caracterização do efeito anti-neoplásico do butirato em duas linhagens celulares de rato derivadas de tumores hepáticos com diferente ní­vel de diferenciação / Characterization of the anti-neoplastic effect of butyrate in two mouse cell lines derived from hepatic tumors with different levels of differentiation

O carcinoma hepatocelular (HCC) é uma neoplasia primária com mau prognóstico e alta taxa de recorrência. Estudos recentes demostram que o HCC pode ser classificado em três subtipos segundo o perfil molecular. Destes subtipos, o HCC pouco diferenciado apresenta pior prognostico. Neste sentido, torna-se de particular interesse o estudo de compostos com efeitos diferenciadores e citotóxicos nas células destas neoplasias pouco diferenciadas. O butirato, um ácido graxo de cadeia curta produzido pela fermentação microbiana da fibra alimentar no intestino, tem demonstrado atividade anti-neoplásica e capacidade moduladora da diferenciação celular em diversos tipos celulares, incluindo linhagens de HCC humano e células progenitoras hepáticas. Assim, objetivou-se neste estudo, caracterizar o efeito do butirato de sódio (NaBu) em duas linhagens de células neoplásicas de rato: uma pouco diferenciada (GP7TB) e a outra, uma linhagem derivada de um HCC diferenciado (JM-1). A linhagem GP7TB mostrou maior resistência ao NaBu (ED50= 7,7 mM) do que as células JM-1 (ED50= 5,2 mM). A redução na viabilidade celular após 72 h de tratamento com NaBu esteve relacionada com a diminuição na proliferação celular e no caso das células GP7TB, de um aumento na apoptose. O tratamento com NaBu induziu alterações morfológicas nas duas linhagens celulares, porém apenas nas células do tipo GP7TB, essas alterações sugerem um processo de diferenciação/transdiferenciação celular. O aumento na expressão de genes envolvidos no controle da pluripotência de células tronco, assim como de alguns marcadores de células tronco, sugere que o NaBu induziu uma reprogramação profunda das células GP7TB. Por outro lado, a redução na expressão de genes relacionados com migração e plasticidade celular assim como de proliferação celular apontam que estas células diminuíram seu potencial invasivo e a capacidade de autorenovação. Embora sejam necessárias análises adicionais para confirmar o efeito observado nos perfis de expressão gênica, os resultados deste estudo sugerem que o NaBu apresenta efeito antineoplásico por meio da redução da proliferação, aumento da apoptose e modulação da expressão de genes associados com a transição epitéliomesenquimal em células com características tronco tumorais

Hepatocellular carcinoma (HCC) is a primary neoplasia with poor prognosis and high recurrence rate. Recent evidence suggests that HCC can be classified in three different subtypes based on their molecular profile. Among these subtypes, the poorlydifferentiated HCC has the worst prognosis. Therefore, the study of compounds with pro-differentiating and cytotoxic effects on poorly-differentiated neoplastic cells represents a matter of primary concern. Butyrate which is a short-chain fatty acid produced by microbial fermentation in the intestine, has demonstrated anti-neoplastic activity and pro-differentiating potential in several cell types, including, human HCC cell lines and liver progenitor cells. In this study, we aimed to characterize the effect of sodium butyrate (NaBu) on two neoplastic cell lines derived from rats: a poorlydifferentiated cell line (GP7TB) and, a cell line derived from a well-differentiated HCC. GP7TB showed increased resistance to NaBu treatment (ED50= 7.7 mM) compared to JM-1 (ED50= 5.2 mM). The reduction in cell viability observed after 72 h of treatment was explained by a reduction in cell proliferation and, in the case of GP7TB, by increased levels of apoptosis. The NaBu treatment induced morphological alterations in both cell lines. However, only in the case of GP7TB cells, the alterations suggested a differentiation/transdifferentiation process. The up-regulation of genes involved in pluripotency and genes expressing stem cell markers indicated that NaBu triggered a deep reprogramming of GP7TB cells. Besides, a down-regulation in the expression of genes related with cell migration and plasticity suggested that these cells reduced their invasive potential and their self-renewal capacity. Additional analyses are necessary to confirm the observed effect on gene expression profiles. However, the results of this study suggest that NaBu exert anti-neoplastic effects through apoptosis, reduction of cell proliferation and downregulation of genes associated with epithelial-mesenchymal transition of cancer stem-like cells

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

Background: Biohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated. Results: The methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15­20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P b 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P b 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield. Conclusion: Preventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.

Combination therapy with butyrate and docosahexaenoic acid for keloid fibrogenesis: an in vitro study

Abstract: Background: A single, effective therapeutic regimen for keloids has not been established yet, and the development of novel therapeutic approaches is expected. Butyrate, a short-chain fatty acid, and docosahexaenoic acid (DHA), a ω-3 polyunsaturated fatty acid, play multiple anti-inflammatory and anticancer roles via their respective mechanisms of action. Objective: In this study, we evaluated the antifibrogenic effects of their single and combined use on keloid fibroblasts. Methods: Keloid fibroblasts were treated with butyrate (0-16 mM) and/or DHA (0-100 µM) for 48 or 96 h. Results: Butyrate inhibited cell proliferation, and α-smooth muscle actin (α-SMA) and type III collagen expressions, with inhibition of the transforming growth factor (TGF)-β1 and TGF-β type I receptor expressions and increased prostaglandin E2 with upregulation of cyclooxygenase-1 expression with induction of histone acetylation. DHA inhibited α-SMA, type III collagen, and TGF-β type I receptor expressions. Then, the butyrate/DHA combination augmented the antifibrogenic effects, resulting in additional inhibition of α-SMA, type I and III collagen expressions, with strong disruption of stress fiber and apoptosis induction. Moreover, the butyrate/DHA combination inhibited the cyclooxygenase-2 expression, suggesting stronger anti-inflammatory effect than each monotherapy. Study limitations: Activation in keloid tissue is affected not only by fibroblasts but also by epithelial cells and immune cells. Evaluation of the effects by butyrate and DHA in these cells or in an in vivo study is required. Conclusion: This study demonstrated that butyrate and docosahexaenoic acid have antifibrogenic effects on keloid fibroblasts and that these may exert therapeutic effects for keloid.

Bacteria in combination with fertilizers promote root and shoot growth of maize in saline-sodic soil

Salinity is the leading abiotic stress hampering maize (Zea mays L.) growth throughout the world, especially in Pakistan. During salinity stress, the endogenous ethylene level in plants increases, which retards proper root growth and consequent shoot growth of the plants. However, certain bacteria contain the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which converts 1-aminocyclopropane-1-carboxylic acid (an immediate precursor of ethylene biosynthesis in higher plants) into ammonia and α-ketobutyrate instead of ethylene. In the present study, two Pseudomonas bacterial strains containing ACC-deaminase were tested separately and in combinations with mineral fertilizers to determine their potential to minimize/undo the effects of salinity on maize plants grown under saline-sodic field conditions. The data recorded at 30, 50 and 70 days after sowing revealed that both the Pseudomonas bacterial strains improved root and shoot length, root and shoot fresh weight, and root and shoot dry weight up to 34, 43, 35, 71, 55 and 68%, respectively, when applied without chemical fertilizers: these parameter were enhanced up to 108, 95, 100, 131, 100 and 198%, respectively, when the strains were applied along with chemical fertilizers. It can be concluded that ACC-deaminase Pseudomonas bacterial strains applied alone and in conjunction with mineral fertilizers improved the root and shoot growth of maize seedlings grown in saline-sodic soil.

Entre o campo e o laboratório: a dinâmica de produção de conhecimento no Ambulatório de Menopausa do Caism/Unicamp / Between the country and the laboratory: the dynamics of the production of knowledge at the Menopause Outpatients Clinic at Caism, Unicamp

Este estudo investiga as práticas de produção de conhecimento sobre a menopausa no Caism/Unicamp, centro de referência para políticas públicas em saúde da mulher. Foram realizadas observações de consultas ginecológicas, entrevistas com mulheres e médicos e observação de reuniões de apoio psicológico, buscando identificar os discursos que circulam no lugar e o processo de alistamento de diferentes atores para que os conhecimentos ali produzidos alcancem credibilidade e “viajem” além dos limites do hospital-escola, tornando-se “universais”. A análise baseia-se nos “estudos localistas”, alinhados aos estudos sociais de ciência e tecnologia.

This study investigates the practices involved in the production of knowledge about menopause at Caism, Unicamp, a reference center for public policies for women’s health. Gynecological appointments and psychological support meetings were observed, and women and doctors were interviewed in order to identify what discourse circulates there and how different actors are brought in to ensure that the knowledge produced attains credibility and “travels” beyond the boundaries of the teaching hospital to become “universal”. The analysis is based on localized studies aligned with social studies of science and technology.

Endophytic fungi producing of esterases: evaluation in vitro of the enzymatic activity using pH indicator

A sensitive and efficient colorimetric method was optimized for detection of esterase enzymes produced by endophytic fungi for development of High-Throughput Screening (HTS). The fungi were isolated and obtained previously from plant species of Cerrado and Atlantic Forest located in areas of environmental preservation in the State of Sao Paulo / Brazil, as part of the project "Chemical and biological prospecting endophytic fungi associated to plant species of Cerrado and Atlantic Forest". The compounds ethyl butyrate, ethyl acetate and methyl propionate were used as standards esters which were hydrolyzed by extracellular enzyme from endophytic fungi (EC. 3.1.1.1 -carboxylesterases) for production of carboxylic acids. Thus, the reduction of the pH increases the protonated indicator concentration (bromothymol blue), changing the color of the reaction medium (from blue to yellow), that can be observed and measured by spectrophotometry at 616 nm. The methodology with acid-base indicator was performed on 13 microorganisms, aiming Periconia atropurpurea asapotential source of esterase for biotransformation of short chain esters. The results also evidenced that this methodology showed to be efficient, fast, cheap, having low consumption of reagents and easy development, and can be applied to screen carboxylic-ester hydrolases in a large number of microorganisms.

Novel organic solvent-tolerant esterase isolated by metagenomics: insights into the lipase/esterase classification / Nueva esterasa tolerante a los solventes orgánicos aislada por metagenómica: ideas sobre la clasificación de las esterasas/lipasas

In order to isolate novel organic solvent-tolerant (OST) lipases, a metagenomic library was built using DNA derived from a temperate forest soil sample. A two-step activity-based screening allowed the isolation of a lipolytic clone active in the presence of organic solvents. Sequencing of the plasmid pRBest recovered from the positive clone revealed the presence of a putative lipase/esterase encoding gene. The deduced amino acid sequence (RBest1) contains the conserved lipolytic enzyme signature and is related to the previously described OST lipase from Lysinibacillus sphaericus 205y, which is the sole studied prokaryotic enzyme belonging to the 4.4 a/ß hydrolase subgroup (abH04.04). Both in vivo and in vitro studies of the substrate specificity of RBest1, using triacylglycerols or nitrophenyl-esters, respectively, revealed that the enzyme is highly specific for butyrate (C4) compounds, behaving as an esterase rather than a lipase. The RBest1 esterase was purified and biochemically characterized. The optimal esterase activity was observed at pH 6.5 and at temperatures ranging from 38 to 45 °C. Enzymatic activity, determined by hydrolysis of p-nitrophenyl esters, was found to be affected by the presence of different miscible and non-miscible organic solvents, and salts. Noteworthy, RBest1 remains significantly active at high ionic strength. These findings suggest that RBest1 possesses the ability of OST enzymes to molecular adaptation in the presence of organic compounds and resistance of halophilic proteins.

Con el fin de aislar nuevas variantes de lipasas tolerantes a solventes organicos (OST), se construyo una libreria metagenomica a partir de ADN obtenido de una muestra de suelo de bosque templado. A traves de un monitoreo en dos etapas, basado en la deteccion de actividades, se aislo un clon con actividad lipolitica en presencia de solventes organicos. La secuenciacion del plasmido pRBest recuperado del clon positivo revelo la presencia de un gen codificante de una hipotetica lipasa/esterasa. La secuencia deducida de amino acidos (RBest1) contiene los motivos conservados de enzimas lipoliticas y esta relacionada con la lipasa OST previamente descrita de Lysinibacillus sphaericus 205y, que es la unica enzima procariota estudiada perteneciente al subgrupo 4.4 de a/ß hidrolasas (abH4.04). Estudios in vivo e in vitro sobre la especificidad de sustratos de RBest1, utilizando triacil-gliceroles o p-nitrofenil-esteres, respectivamente, revelaron que la enzima es altamente especifica para compuestos butiricos (C4), comportandose como una esterasa y no como una lipasa. La esterasa RBest1 fue purificada y caracterizada bioquimicamente. La actividad optima de esterasa fue observada a pH 6,5 y las temperaturas optimas fueron entre 38 y 45 °C. Se establecio que la actividad enzimatica, determinada por hidrolisis de p-nitrofenil esteres, es afectada en presencia de diferentes solventes organicos miscibles y no miscibles, y tambien sales. Notoriamente, RBest1 permanece significativamente activa a elevadas fuerzas ionicas. Estos hallazgos sugieren que RBest1 posee la capacidad de las enzimas OST de la adaptacion molecular en presencia de compuestos organicos, asi como la resistencia de las proteinas halofilas.

Efficacy of the dietary histone deacetylase inhibitor butyrate alone or in combination with vitamin A against proliferation of MCF-7 human breast cancer cells

The combined treatment with histone deacetylase inhibitors (HDACi) and retinoids has been suggested as a potential epigenetic strategy for the control of cancer. In the present study, we investigated the effects of treatment with butyrate, a dietary HDACi, combined with vitamin A on MCF-7 human breast cancer cells. Cell proliferation was evaluated by the crystal violet staining method. MCF-7 cells were plated at 5 x 10(4) cells/mL and treated with butyrate (1 mM) alone or combined with vitamin A (10 µM) for 24 to 120 h. Cell proliferation inhibition was 34, 10 and 46% following treatment with butyrate, vitamin A and their combination, respectively, suggesting that vitamin A potentiated the inhibitory activities of butyrate. Furthermore, exposure to this short-chain fatty acid increased the level of histone H3K9 acetylation by 9.5-fold (Western blot), but not of H4K16, and increased the expression levels of p21WAF1 by 2.7-fold (Western blot) and of RARβ by 2.0-fold (quantitative real-time PCR). Our data show that RARβ may represent a molecular target for butyrate in breast cancer cells. Due to its effectiveness as a dietary HDACi, butyrate should be considered for use in combinatorial strategies with more active retinoids, especially in breast cancers in which RARβ is epigenetically altered.

Efeitos do butirato nos níveis de peroxidação lipídica em células da mucosa cólica sem trânsito fecal: estudo experimental em ratos / Effects of butyrate on levels of lipid peroxidation in cells of the colonic mucosa without fecal stream: experimental study in rats

Os ácidos graxos de cadeia curta (AGCC) representam o principal substrato energético para células da mucosa cólica. A derivação intestinal, reduzindo suprimento de AGCC, responsabiliza-se pela colite de exclusão (CE). Aplicação retal de butirato tem sido eficaz no tratamento da doença. Então, o objetivo deste estudo foi avaliar os níveis de lipoperoxidação na mucosa cólica, após aplicação de butirato, em modelo de CE. Vinte seis ratos Wistar foram submetidos à colostomia proximal e fístula mucosa distal. Os animais foram divididos em dois grupos segundo sacrifício ser realizado em duas ou quatro semanas. Cada grupo foi subdividido em dois subgrups segundo intervenção com soro fisiológico ou butirato. O diagnóstico de CE foi estabelecido por estudo histopatológico e os níveis de lipoperoxidação pelos níveis de malondialdeído (MDA). Utilizaram-se os testes de Mann-Whitney e Kruskal-Wallis (significantes quando p<0,05). Após duas semanas, os níveis de MDA foram menores nos segmentos sem trânsito nos animais irrigados com butirato (p=0,006); porém, após quatro semanas foram semelhantes (p=0,08). No cólon sem trânsito irrigado com butirato, os níveis de MDA aumentaram com o tempo de exclusão (p=0,02); enquanto no cólon com trânsito não se modificaram (p=0,86). O butirato reduz os níveis de MDA na mucosa cólica sem trânsito fecal, após duas semanas de derivação; entretanto, a irrigação isolada não é capaz de reduzir os níveis de lipoperoxidação das células mucosas com o progredir do tempo de exclusão intestinal.

The short-chain fatty acids (SCFA) are the main energy substrate for the cells of the colonic mucosa. Diversion of the fecal stream reducing the supply of SCFA is responsible for diversion colitis (DC). Rectal application of butyrate has been demonstrated effective in the treatment of the disease. So the aim of this study was to evaluate the levels of lipid peroxidation in the colon mucosa after application of butyrate in model of DC. Twenty-six rats were submitted to proximal colostomy and distal mucous fistula. The animals were divided into two groups according sacrifice carried out in two or four weeks. Each group was divided into two subgroups according to intervention with saline solution or butyrate. The diagnosis of colitis was established by histopathology and the levels of lipid peroxidation by tissue levels of malondialdehyde (MDA). We used the Mann-Whitney and Kruskal-Wallis, establishing a significance level of 5 percent (significant with p<0.05). After two weeks, the levels of MDA were lower in the segments without fecal stream of animals irrigated with butyrate (p=0.006), but after four weeks were similar (p=0.08). In the colon without fecal stream irrigated with butyrate, MDA levels increased with the time (p=0.02), while in segments with fecal stream MDA not changed. (p=0.86). Butyrate reduces the levels of MDA in the colonic mucosa without fecal stream, after two weeks of derivation. However, the irrigation with the substance is not able to reduce the lipid peroxidation of mucosal cells with increasing time of intestinal exclusion.

Sodium butyrate does not decrease the evolution of precancerous lesions in rats / Butirato de sódio não diminui a evolução de lesões pré-neoplásicas em ratos

PURPOSE: To evaluate the preventive effect of sodium butyrate in the appearance of aberrant crypt foci (ACF) in rats after induction with the carcinogen 1,2-dimethylhydrazine (DMH). METHODS: Forty Wistar rats were separated into four groups (n=10) distributed as follows: control 1, control 2, butyrate 1 and butyrate 2. The groups control 1 and butyrate 1 remained under experimentation for 4 weeks, while the groups control 2 and butyrate 2 remained for 8 weeks. In the first four weeks, the animals of the control groups received water ad libitum and the animals of the butyrate groups received a sodium butyrate solution (3.4 percent) ad libitum. Injections of the drug 1,2-dimethylhydrazine were applied during the two first weeks of the experiment in all the animals, concurrently with the application of sodium butyrate. The large intestine of the animals was removed, for the analysis of the ACF and of the content of polyamines. The animal feces were collected for the analysis of the SCFA profile. RESULTS: The spermidine presented a higher concentration in the group butyrate 2 in comparison to the group control 2. There was a significant difference in the concentration value (µmol/mL) of acetate in comparison to the groups control 2 and butyrate 2. CONCLUSION: The use of sodium butyrate together with the induction of colorectal cancer was not effective in the prevention of the disease progression.

OBJETIVO: Avaliar o efeito preventivo do butirato de sódio no surgimento de focos de cripta aberrante (FCA) em ratos após a indução com o carcinógeno 1,2-dimetilhidrazina. MÉTODOS: Quarenta ratos foram divididos em quatro grupos, com dez animais em cada. Os grupos controle 1 e butirato 1 ficaram em experimentação por 4 semanas e os grupos controle 2 e butirato 2 por oito semanas. Nas primeiras quatro semanas, os animais dos grupos controle receberam água ad libitum e os animais dos grupos butirato receberam solução de butirato de sódio (3,4 por cento) ad libitum. Em todos os animais foram aplicadas quatro injeções subcutâneas da droga 1,2-dimetilhidrazina nas duas primeiras semanas, concomitante a administração do butirato de sódio. Foi retirado o intestino grosso dos animais, para análise dos FCA e do teor de poliaminas. As fezes dos animais foram recolhidas para análise do perfil de AGCC. RESULTADOS: A espermidina apresentou maior concentração no grupo butirato 2 em relação ao grupo controle 2. Foi encontrada diferença significativa no valor da concentração de acetato quando comparado os grupos controle 2 e butirato 2. CONCLUSÃO: A utilização do butirato de sódio concomitante à indução do câncer colorretal não se mostrou efetiva na prevenção da progressão da doença.

Efeito dos ácidos naftaleno acético e indolilbutírico no enraizamento de estacas de jambolão [Syzygium cumini (L. ) Skeels] / Effect of naphthaleneacetic acid and indolebutyric acid on rooting of jambul [Syzygium cumini (L. ) Skeels] cuttings

O jambolão propaga-se normalmente por sementes o que acarreta variabilidade nas plantas descendentes e um problema quando o objetivo é a formação de pomar comercial. O desenvolvimento de protocolo de propagação vegetativa por meio da estaquia possibilitaria a reprodução de todas as características da planta matriz, uniformidade nas populações e facilidade de propagação. O presente trabalho teve por objetivo avaliar o efeito dos ácidos naftaleno acético (ANA) e indolilbutírico (AIB) no enraizamento de estacas de jambolão. Estacas da região mediana dos ramos foram confeccionadas com 12 cm de comprimento, cortadas em bisel na base e reto acima da última gema axilar, mantendo-se um par de folhas reduzidas à metade. As bases das estacas foram imersas por 10 segundos em soluções aquosas contendo ANA ou AIB nas concentrações de 0, 500, 1.000 e 1.500 mg L-1. Para o plantio foram utilizadas bandejas plásticas contendo areia de granulometria média. As estacas foram mantidas em casa-de-vegetação com nebulização intermitente e após 120 dias do plantio, foram avaliadas as variáveis: porcentagem de estacas enraizadas, com calos, vivas (não enraizadas e sem calos) e mortas, comprimento das três maiores raízes (cm) e número de raízes formadas por estaca. Os melhores resultados de enraizamento foram verificados com 1.000 mg L-1 para ambos os fitorreguladores testados. A porcentagem de enraizamento foi ligeiramente superior com a utilização de ANA quando comparada ao AIB.

Jambul usually propagates by seeds, which causes variability in the descendant plants and represents a problem in the formation of commercial orchards. The development of a protocol for vegetative propagation by cuttings would enable the reproduction of all features of the Mother plant, uniformity in populations and easy propagation. The aim of this work was to evaluate the effect of naphthaleneacetic acid (NAA) and indolebutyric acid (IBA) on rooting of jambul cuttings. Twelve-cm-long cuttings from the median region of branches were prepared through bevel cut in the base and right cut above the last axillary bud, keeping one pair of halved leaves. Cutting bases were immersed for 10s in aqueous solutions containing NAA or IBA at 0, 500, 1000 and 1500 mg L-1 concentrations. Plastic trays containing medium sand were used in the planting. The cuttings were kept in a greenhouse under intermittent nebulization and, at 120 days after planting, the following variables were evaluated: percentage of rooted, with calluses, alive (not-rooted and without calluses) and dead cuttings; length of the three largest roots (cm); and number of roots per cutting. The best rooting was observed by using 1000 mg L-1 of both tested plant growth regulators. Rooting percentage was slightly higher under NAA relative to IBA.

Estaquia de Ginkgo biloba L. utilizando três substratos / Ginkgo biloba L. cutting using three substrates

Ginkgo biloba é arbórea, decídua, cuja folhagem se torna amarelada no outono antes da queda das folhas, o que a torna valorizada em jardinagem. A estaquia é um método de propagação vegetativa baseado na capacidade das células em retomarem o processo de divisão celular, formando raízes em estacas destacadas de ramos provenientes de plantas matrizes. O presente trabalho teve como objetivos verificar a influência de diferentes substratos, assim como, a aplicação da auxina sintética o ácido indol butírico (AIB) no enraizamento de estacas de Ginkgo biloba. No inverno de 2005, ramos foram coletados e transportados até o Laboratório de Macropropagação, onde foram confeccionadas estacas sem folhas, com 10-12 cm de comprimento. Os tratamentos com regulador vegetal (T) foram T1- 0 mg L-1 AIB em solução; T2- 4000 mg L-1 AIB em solução; T3- 8000 mg L-1 AIB em solução; T4- 0 mg kg-1 AIB em talco; T5- 4000 mg kg-1 AIB em talco e T6- 8000 mg kg-1 AIB em talco. Para cada tratamento foram utilizados três diferentes substratos (S), S1- areia, S2- fibra de casca de coco (coxim) e S3- casca de arroz carbonizada. Após 120 dias da instalação, foram avaliadas as porcentagens de estacas enraizadas, vivas, com calos e mortas; o número de raízes por estaca e o comprimento das três maiores raízes por estaca. Os melhores resultados no enraizamento foram obtidos com estacas tratadas com 4000 e 8000 mg kg-1 AIB em talco, utilizando o coxim como substrato (45,00 e 46,25 por cento de enraizamento, respectivamente).

Ginkgo biloba is an arboreal and deciduous species, the foliage of which becomes yellowish in the autumn, before leaf drop, increasing its value for gardening. Cutting is a method of vegetative propagation based on the capacity of cells to recover the cell division process, originating roots in cuttings detached from branches of stock plants. This study aimed to verify the influence of different substrates, as well as the application of the synthetic auxin indole-3-butyric acid (IBA) in Ginkgo biloba cutting rooting. In the winter of 2005, branches were collected and sent to the Macropropagation Lab, where cuttings of 10-12cm length were made without leaves. The treatments with plant growth regulator (T) were T1- 0 mg L-1 IBA solution, T2- 4000 mg L-1 IBA solution, T3- 8000 mg L-1 IBA solution, T4- 0 mg kg-1 IBA in talc, T5- 4000 mg kg-1 IBA in talc, T6- 8000 mg kg-1 IBA in talc. Each treatment was planted in three substrates (S), S1- sand, S2- coir and S3- carbonized rice hull. After 120 days, the percentages of cuttings that were rooted, alive, with callus and dead were evaluated, besides the number of roots per cutting and the length of the three highest roots per cutting. The best results regarding rooting were obtained for cuttings treated with 4000 and 8000 mg kg-1 IBA in talc, by using coir as substrate (45.00 and 46.25 percent rooting, respectively).

Oral administration of sodium butyrate reduces chemically-induced preneoplastic lesions in experimental carcinogenesis

OBJECTIVE: The objective was to assess the effects of oral administration of sodium butyrate on colon carcinogenesis. METHODS: Carcinogenesis in adult male Wistar rats was induced with 1.2-dimethylhydrazine injections at a dose of 40mg/kg of body weight. A solution of sodium butyrate (3.4 percent) was given ad libitum for 4 weeks (butyrate group, n=16) instead of water (control group, n=9). Rats were killed 17 weeks after 1.2-dimethylhydrazine administration. Aberrant crypt foci and expression of the messenger ribonucleic acid (mRNA) of cyclins D1 and E were quantified in the colon. Alterations in the fatty acid profile of the colon, liver, intra-abdominal fat and feces were also analyzed. RESULTS: A significant decrease in aberrant crypt foci was found in the group taking butyrate. No differences were found between the groups in the mRNA expression of cyclins D1 and E. Nevertheless, butyrate intake decreased the content of stearic and oleic acids in the intra-abdominal fat and docosahexaenoic acid in the liver. Moreover, these rats presented higher percentages of linoleic acid in the intra-abdominal fat than control rats. CONCLUSION: The data indicate that butyrate use in rats reduced preneoplastic lesions and changes in the intra-abdominal fat and fatty acid profile of the liver, commonly found in colon carcinogenesis.

OBJETIVO: Avaliar o efeito da administração oral de butirato de sódio na carcinogênese do cólon. MÉTODOS: A carcinogênese em ratos Wistar foi induzida com injeções de 1,2-dimetilhidrazina na dose de 40mg/kg de peso corporal. A solução de butirato de sódio (3,4 por cento) foi oferecida ad libitum por 4 semanas (grupo butirato, n=16), em substituição à água (grupo controle, n=9). Os ratos foram sacrificados na 17ª semana após tratamento com a 1,2-dimetilhidrazina. Focos de criptas aberrantes e a expressão dos genes para o ácido ribonucléico mensageiro (RNAm) das ciclinas D1 e E foram quantificadas no cólon. Alterações no perfil de ácidos graxos do cólon, no fígado, na gordura intra-abdominal e nas fezes foram analisadas. RESULTADOS: Uma significante diminuição nos focos de criptas aberrantes foi encontrada no grupo que recebeu butirato. Nenhuma diferença foi encontrada na expressão do RNAm das ciclinas D1 e E entre os grupos. Contudo, a ingestão de butirato diminuiu a quantidade de ácido esteárico e ácido oléico na gordura intra-abdominal e do ácido docosahexanóico no fígado. Além disso, o grupo butirato apresentou maior percentual de ácido linoléico na gordura intra-abdominal, comparado com os ratos do grupo controle. CONCLUSÃO: Os dados indicam que o uso de butirato reduz lesões pré-neoplásicas em ratos e diminui as alterações no perfil de ácidos graxos da gordura intra-abdominal e do fígado, comumente encontradas na carcinogênese colônica.

Avaliação das condições reacionais para a síntese enzimática do butirato de butila empregando lipase de Candida rugosa / Evaluation of the reaction conditions for the enzymatic synthesis of the butyl butyrate using lipase from Candida rugosa

Lipase de Candida rugosa na forma livre foi usada na síntese de butirato de butila pela esterificação direta de n-butanol com ácido butírico. Um planejamento fatorial completo 24 foi empregado para determinar a influência da razão molar do álcool para ácido (1:0,5­2,5), concentração de agente dessecante (0­20 por cento), concentração de enzima (40­80 mg) e temperatura de incubação (30­60 C) no rendimento de esterificação. A concentração de agente dessecante foi o fator mais significativo na esterificação, sendo sua influência negativa. O progresso da esterificação foi favorecido para substratos contendo ácido em excesso, mesmo em baixa concentrações de enzima (40 mg), sendo a conversão de 50 por cento...

Estudo do efeito do leite fermentado contendo Lactobacillus casei Shirota na microbiota intestinal de crianças sob terapia antimicrobiana / Study of the effect of fermented milk contend Lactobacillus casei Shirote in the intestinal microflora of children under antimicrobial therapy

O tratamento antimicrobiano pode destruir o equilíbrio da microbiota gastrintestinal, podendo induzir sintomas clínicos, principalmente a diarréia. A influência de Lactobacillus casei Shirota sobre a microbiota intestinal foi avaliada em um estudo prospectivo, randomizado, duplo-cego e controlado. Sessenta e três crianças hospitalizadas, com idade de 2-14 anos, sob tratamento com antibióticos ß-lactâmicos foram randomizadas para receber o leite fermentado por L. casei Shirota, `10 POT. 8-9ï UFC/mL, ou o placebo durante o tratamento antimicrobiano. As amostras de fezes foram colhidas antes da administração do leite fermentado, durante o tratamento antibiótico, e uma semana após o término do tratamento com o antimicrobiano e a ingestão do leite fermentado...