Atividades Biológicas De Xilana De Sabugo De Milho

The corn cob is an agricultural by-product still little used, this in part due to the low knowledge of the biotechnological potential of their molecules. Xylan from corn cobs (XSM) is a polysaccharide present in greater quantity in the structure of plant and its biotechnology potential is little known. This study aimed to the extraction, chemical characterization and evaluation of biological activities of xylan from corn cobs. To this end, corncobs were cleaned, cut, dried and crushed, resulting in flour. This was subjected to a methodology that combines the use of alkaline conditions with waves of ultrasound. After methanol precipitation, centrifugation and drying was obtained a yield of 40% (g/g flour). Chemical analysis indicated a high percentage of polysaccharides in the sample (60%) and low contamination by protein (0.4%) and phenolic compounds (> 0.01%). Analysis of monosaccharide composition indicated the presence of xylose:glucose:arabinose:galactose:mannose:glucuronic acid in a molar ratio 50:20:15:10:2.5:2.5. The presence of xylan in the sample was confirmed by nuclear magnetic resonance (¹H and ¹³C) and infrared spectroscopy (IR). Tests were conducted to evaluate the antioxidant potential of XSM. This showed a total antioxidant capacity of 48.45 EAA/g sample. However, did not show scavenging activity of superoxide and hydroxyl radical and also reducing power. But, showing a high capacity chelating iron ions with 70% with about 2 mg/mL. The ability to XSM to influence cell proliferation in culture was also evaluated. This polymer did not influence the proliferation of normal fibroblast cells (3T3), however, decreased the rate of proliferation of tumor cells (HeLa) in a dose-dependent, reaching an inhibition of about 50% with a concentration around 2 mg/mL. Analyzing proteins related to cell death, by immunoblotting, XSM increases the amount of Bax, Bcl-2 decrease, increase cytochrome c and AIF, and reduce pro-caspase-3, indicating the induction of cell death induced apoptosis dependent and independent of caspase. XSM did not show anticoagulant activity in the PT test. However, the test of activated partial thromboplastin time (aPTT), XSM increased clotting time at about 5 times with 600 μg of sample compared with the negative control. The presence of sulfate on the XSM was discarded by agarose gel electrophoresis and IR. After carboxyl-reduction of XSM the anticoagulant activity decreased dramatically. The data of this study demonstrate that XSM has potential as antioxidant, antiproliferative and anticoagulant compound. Future studies to characterize these activities of XSM will help to increase knowledge about this molecule extracted from corn and allow their use in functional foods, pharmaceuticals and chemical industries.

Desenvolvimento de uma formulação cólon específica visando o tratamento da colite ulcerativa

Micro and nanoparticulate systems as drug delivery carriers have achieved successful therapeutic use by enhancing efficacy and reducing toxicity of potent drugs. The improvement of pharmaceutical grade polymers has allowed the development of such therapeutic systems. Microencapsulation is a process in which very thin coatings of inert natural or synthetic polymeric materials are deposited around microsized particles of solids or around droplets. Products thus formed are known as microparticles. Xylan is a natural polymer abundantly found in nature. It is the most common hemicellulose, representing more than 60% of the polysaccharides existing in the cell walls of corn cobs, and is normally degraded by the bacterial enzymes present in the colon of the human body. Therefore, this polymer is an eligible material to produce colon-specific drug carriers. The aim of this study was to evaluate the technological potential of xylan for the development of colon delivery systems for the treatment of inflammatory bowel diseases. First, coacervation was evaluated as a feasible method to produce xylan microcapsules. Afterwards, interfacial cross-linking polymerization was studied as a method to produce microcapsules with hydrophilic core. Additionally, magnetic xylan-coated microcapsules were prepared in order to investigate the ability of producing gastroresistant systems. Besides, the influence of the external phase composition on the production and mean diameter of microcapsules produced by interfacial cross-linking polymerization was investigated. Also, technological properties of xylan were determined in order to predict its possible application in other pharmaceutical dosage forms

Novos sistemas de liberação de fármacos à base de xilana

The aim of this work was to perform the extraction and characterization of xylan from corn cobs and prepare xylan-based microcapsules. For that purpose, an alkaline extraction of xylan was carried out followed by the polymer characterization regarding its technological properties, such as angle of repose, Hausner factor, density, compressibility and compactability. Also, a low-cost and rapid analytical procedure to identify xylan by means of infrared spectroscopy was studied. Xylan was characterized as a yellowish fine powder with low density and poor flow properties. After the extraction and characterization of the polymer, xylan-based microcapsules were prepared by means of interfacial crosslinking polymerization and their characterization was performed in order to obtain gastroresistant multiparticulate systems. This work involved the most suitable parameters of the preparation of microcapsules as well as the study of the process, scale-up methodology and biological analysis. Magnetic nanoparticles were used as a model system to be encapsulated by the xylan microcapsules. According to the results, xylan-based microcapsules were shown to be resistant to several conditions found along the gastrointestinal tract and they were able to avoid the early degradation of the magnetic nanoparticles

Nanotechnological delivery systems for the oral administration of active molecules: Polymeric microparticles and microemulsions applied to anti-inflammatory and anti-infectious drugs

This thesis was devoted to the development of innovative oral delivery systems for two different molecules. In the first part, microparticles (MPs) based on xylan and Eudragit® S- 100 were produced and used to encapsulate 5-aminosalicylic acid for colon delivery. Xylan was extracted from corn cobs and characterized in terms of its physicochemical, rheological and toxicological properties. The polymeric MPs were prepared by interfacial cross-linking polymerization and spray-drying and characterized for their morphology, mean size and distribution, thermal stability, crystallinity, entrapment efficiency and in vitro drug release. MPs with suitable physical characteristics and satisfactory yields were prepared by both methods, although the spray-dried systems showed higher thermal stability. In general, spraydried MPs would be preferable systems due to their thermal stability and absence of toxic agents used in their preparation. However, drug loading and release need to be optimized. In the second part of this thesis, oil-in-water microemulsions (O/W MEs) based on mediumchain triglycerides were formulated as drug carriers and solubility enhancers for amphotericin B (AmB). Phase diagrams were constructed using surfactant blends with hydrophiliclipophilic balance values between 9.7 and 14.4. The drug-free and drug-loaded MEs presented spherical non-aggregated droplets around 80 and 120 nm, respectively, and a low polydispersity index. The incorporation of AmB was high and depended on the volume fraction of the disperse phase. These MEs did not reduce the viability of J774.A1 macrophage-like cells for concentrations up to 25 μg/mL of AmB. Therefore, O/W MEs based on propylene glycol esters of caprylic acid may be considered as suitable delivery systems for AmB

Micropartículas poliméricas à base de xilana e Eudragit® S-100 contendo mesalazina visando à liberação cólon-específica

Colon-specific drug delivery systems have attracted increasing attention from the pharmaceutical industry due to their ability of treating intestinal bowel diseases (IBD), which represent a public health problem in several countries. In spite of being considered a quite effective molecule for the treatment of IBD, mesalazine (5-ASA) is rapidly absorbed in the upper gastrointestinal tract and its systemic absorption leads to risks of adverse effects. The aim of this work was to develop a microparticulate system based on xylan and Eudragit® S- 100 (ES100) for colon-specific delivery of 5-ASA and evaluate the interaction between the polymers present in the systems. Additionaly, the physicochemical and rheological properties of xylan were also evaluated. Initially, xylan was extracted from corn cobs and characterized regarding the yield and rheological properties. Afterwards, 10 formulations were prepared in different xylan and ES100 weight ratios by spray-drying the polymer solutions in 0.6N NaOH and phosphate buffer pH 7.4. In addition, 3 formulations consisting of xylan microcapsules were produced by interfacial cross-linking polymerization and coated by ES100 by means of spray-drying in different polymer weight ratios of xylan and ES100. The microparticles were characterized regarding yield, morphology, homogeneity, visual aspect, crystallinity and thermal behavior. The polymer interaction was investigated by infrared spectroscopy. The extracted xylan was presented as a very fine and yellowish powder, with mean particle size smaller than 40μm. Regarding the rheological properties of xylan, they demonstrated that this polymer has a poor flow, low density and high cohesiveness. The microparticles obtained were shown to be spherical and aggregates could not be observed. They were found to present amorphous structure and have a very high thermal stability. The yield varied according to the polymer ratios. Moreover, it was confirmed that the interaction between xylan and ES100 occurs only by means of physical aggregation

Requirement for cobalamin by Salmonella enterica serovars Typhimurium, Pullorum, Gallinarum and Enteritidis during infection in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A defective mutant of Salmonella enterica Serovar Gallinarum in cobalamin biosynthesis is avirulent in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of Salmonella Enteritidis and Salmonella Gallinarum in birds by using live vaccine candidate containing attenuated Salmonella Gallinarum mutant strain

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da composição química do substrato no cultivo de Pleurotus florida

Com o objetivo de avaliar a influência da composição química do substrato no cultivo do cogumelo comestível Pleurotus florida, seis resíduos agrícolas foram testados: palha de arroz, palha de feijão, palha de trigo, folha de bananeira, palha de sorgo e sabugo de milho. O substrato palha de feijão apresentou resultados semelhantes para a produção (189,8 g kg-1), eficiência biológica (89,2%) e número de cogumelos (12) à palha de arroz, substrato utilizado tradicionalmente no cultivo de Pleurotus em escala comercial. Não foi possível atribuir apenas a um fator químico as altas produções e eficiências biológicas observadas em palha de arroz e palha de feijão e muito menos para a baixa produção em palha de sorgo (77,8 g kg-1) e sabugo de milho (53,2 g g-1). No geral, substratos com relação C/N em torno de 45 (N = 1,0%), maior conteúdo de cinzas, acrescido de altos teores de P, K, Ca, Mg and Mn foram os melhores para o cultivo de P. florida.

Predição da digestibilidade da matéria seca de rações em coelhos em crescimento

O experimento foi realizado com o objetivo de predizer a digestibilidade da matéria seca (MS) de rações para coelhos em crescimento mediante equações baseadas no conceito de entidade nutritiva ideal. Foram usados 100 coelhos do Grupo Genético Botucatu, com 42 dias de idade. Testaram-se 16 rações contendo alimentos volumosos (papel, sabugo de milho, bagaço de cana ou maravalha de pínus), mais milho, farelo de soja, óleo de soja, caulim e sal comum. Os volumosos foram incluídos em quatro níveis, para se obter 22, 28, 34 ou 40% de fibra em detergente neutro (FDN) e 16% de proteína bruta (PB). O experimento foi realizado em duas fases, em delineamento experimental de blocos ao acaso. Observou-se efeito linear positivo da FDN sobre o consumo de MS. As médias de digestibilidade da MS variaram entre 60,10 e 80,78%. Houve efeito linear negativo da FDN, bem como interação do teor de FDN × volumoso, sobre a digestibilidade aparente da MS. Ao contrário dos componentes fibrosos, a proteína bruta PB e o conteúdo celular comportaram-se como entidade nutritiva ideal. Para predizer a digestibilidade da MS, testaram-se três modelos e o melhor baseou-se no teor de conteúdo celular, com digestibilidade verdadeira fixada em 1; outro no teor de FDN, cuja digestibilidade foi predita mediante uma equação empírica; e o terceiro, na perda endógena de conteúdo celular, considerada proporcional ao consumo de MS. A principal dificuldade na formulação de modelos para predizer a digestibilidade aparente da MS em coelhos é estimar com precisão a digestibilidade da FDN e a perda endógena de conteúdo celular. É necessário avaliar em estudos futuros o uso de maiores níveis de fibra nas rações. Para rações com níveis de fibra situados na amplitude deste estudo, recomenda-se usar o modelo 3, que inclui, além da fibra em detergente neutro, a fibra em detergente ácido e a lignina.

Infecção mista pelo Sugarcane mosaic virus e Maize rayado fino virus provoca danos na cultura do milho no estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do capim-elefante (Pennisetum purpureum Schum.) cultivar taiwan A-148, ensilado com diferentes técnicas de redução de umidade

O objetivo deste trabalho foi avaliar o efeito de diferentes técnicas de redução de umidade sobre a composição química, digestibilidade in vitro da matéria seca (DIVMS) e degradação ruminal da matéria seca da silagem de capim-elefante (Pennisetum purpureum Schum.) cv. Taiwan A-148. Os tratamentos (A, Controle; B, 20,0; C, 30,0; D, 40,0% de sabugo de milho; E, emurchecimento por 12 horas; F, emurchecimento por 24 horas; e G, esmagamento + emurchecimento por 24 horas) foram distribuídos em delineamento inteiramente casualizado com três repetições. Os tratamentos da silagem B e E não foram eficientes em reduzir a umidade excessiva da forragem. O emurchecimento por 24 horas aumentou o teor de MS, sem influir no teor de PB e na DIVMS. Os tratamentos C e D favoreceram o desenvolvimento da população de clostrídeos, aumentaram a concentração de N-NH3 e reduziram a concentração de ácido lático e a DIVMS das silagens. O tratamento G aumentou o teor de MS e reduziu a concentração de N-NH3 e DIVMS. A adiçao de sabugo de milho (tratamentos B, C e D) reduziu a degradação ruminal da MS das silagens. O sabugo de milho reduziu a umidade, mas apresentou efeitos negativos na qualidade da silagem, enquanto o esmagamento e/ou emurchecimento foram procedimentos que mostraram bons resultados em relação à conservação do material ensilado.

Avaliação da silagem de grãos úmidos de milho

This work had the objective of evaluating the effects of different percentages of corn (Zea mays L.) cob on the quality of the silage of high-moisture corn grains. The following treatments were studied: percentages of corn cob in the silage (0, 5, 10, 15 and 20%); period of sampling after opening the silos (zero, two, four and six days). The factorial arrangement 5 × 4 was studied according to a completely randomized block design with three replications. The variables studied were not affected by the sampling period. The buffering capacity and the pH were not affected by the cob, while the percent soluble carbohydrates and amoniacal nitrogen increasing until 1.2 and 1.89 unit percent, respectively. Increasing the amount of corn cobs reduced the contents of DM (from 63.9% to 58.6%), CP (from 10.0% to 7.3%), EE (from 4.87% to 3.92%) and the values of DMIVD (from 90.5% to 79.1%) in the silages and increasing the contents of acid detergente fiber (ADF) (from 3.3% to 12.9%) and neutral detergente fiber (NDF) (from 15.16% to 26.1%). The values of brute energy (BE) were not affected (P>0.01) by the cob corn in the silage.

Participation of genes involved in the process of anaerobic respiration of infection in chickens by salmonella typhimurium

Intestinal pathogens are exposed to various stress conditions during their infectious cycle. Anaerobiosis, one of such hostile condition, is offered by the host within gut and intestinal lumen, where survival, multiplication and entry into intestinal epithelial cells are priority for the invasion of the pathogen. The fumarate reductase (frdABCD), dimethyl sulfoxide (DMSO)-trimethylamine N-oxide (TMAO) reductase (dmsABC), and nitrate reductase (narGHIJ) operons in Salmonella Typhimurium (STM) encode enzymes involved in anaerobic respiration to the electron acceptors fumarate, DMSO, TMAO, and nitrate, respectively. They are regulated in response to nitrate and oxygen availability and changes in cell growth rate. Vitamin B12 (cobalamin) is synthesized by Salmonella Typhimurium only under anaerobic growth conditions used as a cofactor in four known reactions. The deletion of cobS and cbiA genes prevent any form of cobalamin production. In the present study we evaluate the infection of birds by mutants of STM, with the anaerobic respiratory system committed by mutations in the genes: narG, napA, cobS, cbiA, frdA, dmsA, and torC. Virulence was assessed by oral inoculation of groups of one-day-old broilers with 0.1 mL of culture contained 10 8 colony forming units (CFU)/mL or diluted at 10 -3 and 10 -2 of strains mutants of Salmonella Typhimurium. Clinical signs and mortality were recorded over a period of 21 days. In general, the symptoms of chickens infected with the mutant strains were similar to those presenting by control birds. Except for STMNalr cbiA, all showed reduced capacity to cause mortality in comparison with the original strain. The mortality of group of chickens infected with STMNal r △narG, STMNal r △frdA, STMNal r △dmsA and STMNal r △cobS△cbiA showed significant decrease in mortality compared to control group (p<0.05).

Experimental infection of commercial layers with wild or attenuated Salmonella Gallinarum mutant strains: Anatomic pathology, total blood cell count and serum protein levels

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)