RESUMO

During human pregnancy, leukocytes that infiltrate the maternal-fetal interface play a major role in establishing a delicate balance between immune tolerance and functional response and setting the inflammatory process that leads to labor. Here we describe two methods for isolating immune cells from the chorioamniotic membranes (decidua parietalis) and placental blood (decidua basalis) that combine gentle enzymatic digestion, magnetic cell sorting, and density gradient. Isolated leukocytes can be immunophenotypified by flow cytometry, and both isolation methods are compatible with downstream cellular and molecular applications, such as cell culture, transcriptome, and proteome analyses.

Assuntos

RESUMO

Multifunctional peptides, capable of acting on different body systems through multiple mechanisms of action, offer many advantages over monofunctional peptides, including lower adverse side effects and costs. Erythrina edulis (pajuro) is a legume with a large number of high-quality proteins, of which their potential as a source of antioxidant peptides has been recently reported. In this study, the behavior of these proteins under a sequential enzymatic hydrolysis with digestive and microbial enzymes was investigated by evaluating the multi-functionality of the hydrolyzates. The albumin hydrolyzates obtained after the action of pepsin, pancreatin, and Alcalase showed antioxidant, angiotensin-converting enzyme (ACE), α-amylase, α-glucosidase, and dipeptidyl peptidase (DPP)-IV inhibitory activities. The radical scavenging properties of the hydrolyzate could be responsible for the potent protective effects observed in FeSO4-induced neuroblastoma cells. The findings support the role of pajuro protein as an ingredient of functional foods or nutraceuticals for health promotion and the prevention of oxidative stress, hypertension, and metabolic alteration-associated chronic diseases.

RESUMO

BACKGROUND: Horses are hindgut fermenters, and it is therefore important to determine the postgastric nutritive value of their feedstuffs and diets. Moreover, it has been demonstrated in other animal species that the fermentation of diets results in different values than those expected from pure ingredients. Therefore, the general objective of this work is to evaluate the gas production (GP) and volatile fatty acid (VFA) concentration, as well as the associative effects, of mixtures of different forages and concentrated foods, which are representative of the traditional diets of high-performance horses. METHODS: An in vitro gas production experiment was conducted to assess the fermentation of two forages and three concentrates that are typical in horse diets. The combination of 70% of forage and 30% concentrates was also assessed to determine potential associative effects. RESULTS: Concentrates and grains produced higher GP and VFA than forages when evaluated alone. When experimental diets were incubated, GP parameters and VFA concentrations of forage-concentrate mixtures had unexpected differences from the values expected from the fermentation of pure ingredients, suggesting the occurrence of associative effects. CONCLUSIONS: Our results indicate that there is a need to evaluate the fermentation of diets, rather than predicting from the values of pure ingredients.

RESUMO

Objetivo: Analizar genéticamente la resistencia a Isoniacida en cepas de Mycobacterium tuberculosis mediante PCR-RFLP de la región S315T del gen katG. Materiales y métodos: El estudio se realizó a partir de cultivos positivos de Mycobacterium tuberculosis receptados en el Centro de Referencia Nacional de Micobacterias, durante el período 2013 ­ 2014. El ADN extraído fue cuantificado y evaluada su pureza, por espectrofotometría. Para determinar el polimorfismo en la región 315 del gen katG a partir de un producto de amplificación de 630 pb se realizó digestiones con las enzimas de restricción MspI y SatI. Resultados: Del total de 498 cepas analizadas, 215 cepas presentaron características fenotípicas de resistencia a isoniacida (32,6 % monorresistencia, 19,5 % MDR y 47,9 % polirresistencia), 283 cepas eran sensibles. 251 cepas correspondieron a pacientes vírgenes al tratamiento (VT); 174 fueron pacientes antes tratados (AT) y 73 fueron pacientes se encontraban con tratamiento (CT). La mayoría de los casos provenía de la provincia del Guayas (77,2 %). La PCR-RFLP-SatI presentó alto porcentaje de sensibilidad (98,6 %) y especificidad (98,2 %), mientras que con la enzima MspI el porcentaje de sensibilidad fue 88,8 % y 7,4 % de especificidad. Conclusión: La PCR-RFLP-SatI demostró ser específica y económica para la detección de resistencia a isoniacida, proporcionando resultados de forma rápida, la aplicación de esta técnica como apoyo para el diagnóstico permitiría al paciente acceder a un tratamiento más oportuno.

Objective: Genetically analyze the Isoniazid resistance in Mycobacterium tuberculosis cultures by PCR-RFLP of the S315T region of the atG. Materials and methods: The study was carried out in positive cultures of Mycobacterium tuberculosis, received at the National Reference Center of Mycobacteria, during the period 2013-2014. The DNA extracted was quantified and its purity was evaluated by spectrophotometry. To determine the polymorphism in the 315 region of the at G gene, digestions were made with the restriction enzymes MspI and SatI from a 630 bp amplification product. Results: Of 498 culture strains analyzed, 215 strains showed phenotypic characteristics of resistance to Isoniazid (32,6 % monoresistance, 19,5 % MDR and 47,9 % polyresistance) and 283 strains were sensitive. 251 strains corresponded to virgin patients to treatment (VT); 174 were patients before treated (AT) and 73 were patients treated (CT). The majority of cases came from the province of Guayas (77,2 %). The PCR-RFLP- SatI presented a high percentage of sensitivity (98,6 %) and specificity (98,2 %), while with the MspI enzyme the sensitivity percentage was 88,8 % and 7,4 % specificity. Conclusion: The PCR-RFLP SatI proved to be specific and economical for the detection of resistance to isoniazid, providing results quickly, the application of this technique as a support for the diagnosis would allow the patient to access a more timely treatment.

Assuntos

RESUMO

SUMMARY This study analyse ruminal degradation the technique of the nylon bags of dry matter (DM) and crude protein (CP) and the intestinal digestibility of rumen undegraded protein (RUP) by the method of the three stages of raw and roasted soybeans at different temperatures with and without The in situ ruminal degradation were weighed five grams of natural matter in nylon bags incubated 2; 4;8; 16; 24 and 48 hours. Zero time was made the same procedure, except ruminal incubation. The residue of treatment formed a composite sample to determine the DM and CP. The intestinal digestibility was in situ incubation for 16 hours, using the technique of the three stages. The degradability of DM at a passage rate of 5 % / hour, for raw soybean (RS), was 71.94 % and roasted between 52.23 % and 68.78 %. After 16 hours of incubation(RUP) ranged from 32.12 to 67.72% and the intestinal digestibility of 73.21% to 86.02 %. The lowest degradation of DM and CP was in the roasted soybeans at 145ºC for one minute with steeping (STC1). The in vitro intestinal digestibility of raw grains was higher and differed from the toasted, except for the ones toasted at 115 ºC for four minutes with steeping. For STC1, was obtained the lowest protein degradation of 67.72 % RUP, which 52.33 %, more when compare to RS. The toast of soybeans at 145 ºC (STC1) contributed to a lower ruminal degradability of crude protein.

RESUMO Objetivou-se estudar o efeito de diferentes tratamentos térmicos, tempos e procedimentos na degradação ruminal de grãos de soja crus e tostados e sua ação na digestão intestinal da proteína não degradada no rúmen (PNDR) pelo método dos três estágios. Para a degradação ruminal in situ foram pesados cinco gramas de matéria natural em sacos de náilon incubados durante 2; 4; 8; 16; 24 e 48 horas. No tempo zero foi efetuado o mesmo procedimento, excetuando a incubação ruminal. Os resíduos de cada tratamento formaram uma amostra composta para determinar a matéria seca (MS) e proteína bruta (PB). Para a digestibilidade intestinal realizou-se a incubação in situ por 16 horas, usando a técnica dos três estágios. A degradabilidade efetiva da MS com taxa de passagem de 5%/hora para a soja crua (SC) foi de 71,94% e tostada entre 52,23% a 68,78%. Após 16 horas de incubação a PNDR variou de 32,12 a 67,72% e a digestibilidade intestinal de 73,21% a 86,02%. A menor degradação da MS e PB foi da soja tostada a 145°C durante um minuto com steeping (STC1). A digestibilidade intestinal in vitro dos grãos crus foi superior e diferiu dos tostados, exceto a soja tostada a 115°C durante quatro minutos com steeping. A menor degradação proteica foi obtida da STC1de 67,72% da PNDR,52,33% a mais do que à SC. A tostagem dos grãos de soja a 145°C(STC1)contribuiu para uma menor degradabilidade ruminal da proteína bruta.

RESUMO

Objetivou-se estudar o efeito de diferentes tratamentos térmicos, tempos e procedimentos na degradação ruminal de grãos de soja crus e tostados e sua ação na digestão intestinal da proteína não degradada no rúmen (PNDR) pelo método dos três estágios. Para a degradação ruminal in situ foram pesados cinco gramas de matéria natural em sacos de náilon incubados durante 2; 4; 8; 16; 24 e 48 horas. No tempo zero foi efetuado o mesmo procedimento, excetuando a incubação ruminal. Os resíduos de cada tratamento formaram uma amostra composta para determinar a matéria seca (MS) e proteína bruta (PB). Para a digestibilidade intestinal realizou-se a incubação in situ por 16 horas, usando a técnica dos três estágios. A degradabilidade efetiva da MS com taxa de passagem de 5%/hora para a soja crua (SC) foi de 71,94% e tostada entre 52,23% a 68,78%. Após 16 horas de incubação a PNDR variou de 32,12 a 67,72% e a digestibilidade intestinal de 73,21% a 86,02%. A menor degradação da MS e PB foi da soja tostada a 145oC durante um minuto com steeping (STC1). A digestibilidade intestinal in vitro dos grãos crus foi superior e diferiu dos tostados, exceto a soja tostada a 115oC durante quatro minutos com steeping. A menor degradação proteica foi obtida da STC1de 67,72% da PNDR,52,33% a mais do que à SC. A tostagem dos grãos de soja a 145oC(STC1)contribuiu para uma menor degradabilidade ruminal

This study analyse ruminal degradation the technique of the nylon bags of dry matter ( DM) and crude protein (CP ) and the intestinal digestibility of rumen undegraded protein (RUP) by the method of the three stages of raw and roasted soybeans at different temperatures with and without The in situ ruminal degradation were weighed five grams of natural matter in nylon bags incubated 2; 4;8; 16; 24 and 48 hours. Zero time was made the same procedure, except ruminal incubation. The residue of treatment formed a composite sample to determine the DM and CP. The intestinal digestibility was in situ incubation for 16 hours, using the technique of the three stages. The degradability of DM at a passage rate of 5 % / hour, for raw soybean (RS), was 71.94 % and roasted between 52.23 % and 68.78 %. After 16 hours of incubation(RUP) ranged from 32.12 to 67.72% and the intestinal digestibility of 73.21% to 86.02 %. The lowest degradation of DM and CP was in the roasted soybeans at 145ºC for one minute with steeping (STC1). The in vitro intestinal digestibility of raw grains was higher and differed from the toasted, except for the ones toasted at 115 ºC for four minutes with steeping. For STC1, was obtained the lowest protein degradation of 67.72 % RUP, which 52.33 %, more when compare to RS. The toast of soybeans at 145 ºC (STC1) contributed to a lower ruminal degradability of crude protein.

Assuntos

RESUMO

SUMMARY This study analyse ruminal degradation the technique of the nylon bags of dry matter (DM) and crude protein (CP) and the intestinal digestibility of rumen undegraded protein (RUP) by the method of the three stages of raw and roasted soybeans at different temperatures with and without The in situ ruminal degradation were weighed five grams of natural matter in nylon bags incubated 2; 4;8; 16; 24 and 48 hours. Zero time was made the same procedure, except ruminal incubation. The residue of treatment formed a composite sample to determine the DM and CP. The intestinal digestibility was in situ incubation for 16 hours, using the technique of the three stages. The degradability of DM at a passage rate of 5 % / hour, for raw soybean (RS), was 71.94 % and roasted between 52.23 % and 68.78 %. After 16 hours of incubation(RUP) ranged from 32.12 to 67.72% and the intestinal digestibility of 73.21% to 86.02 %. The lowest degradation of DM and CP was in the roasted soybeans at 145ºC for one minute with steeping (STC1). The in vitro intestinal digestibility of raw grains was higher and differed from the toasted, except for the ones toasted at 115 ºC for four minutes with steeping. For STC1, was obtained the lowest protein degradation of 67.72 % RUP, which 52.33 %, more when compare to RS. The toast of soybeans at 145 ºC (STC1) contributed to a lower ruminal degradability of crude protein.

RESUMO Objetivou-se estudar o efeito de diferentes tratamentos térmicos, tempos e procedimentos na degradação ruminal de grãos de soja crus e tostados e sua ação na digestão intestinal da proteína não degradada no rúmen (PNDR) pelo método dos três estágios. Para a degradação ruminal in situ foram pesados cinco gramas de matéria natural em sacos de náilon incubados durante 2; 4; 8; 16; 24 e 48 horas. No tempo zero foi efetuado o mesmo procedimento, excetuando a incubação ruminal. Os resíduos de cada tratamento formaram uma amostra composta para determinar a matéria seca (MS) e proteína bruta (PB). Para a digestibilidade intestinal realizou-se a incubação in situ por 16 horas, usando a técnica dos três estágios. A degradabilidade efetiva da MS com taxa de passagem de 5%/hora para a soja crua (SC) foi de 71,94% e tostada entre 52,23% a 68,78%. Após 16 horas de incubação a PNDR variou de 32,12 a 67,72% e a digestibilidade intestinal de 73,21% a 86,02%. A menor degradação da MS e PB foi da soja tostada a 145°C durante um minuto com steeping (STC1). A digestibilidade intestinal in vitro dos grãos crus foi superior e diferiu dos tostados, exceto a soja tostada a 115°C durante quatro minutos com steeping. A menor degradação proteica foi obtida da STC1de 67,72% da PNDR,52,33% a mais do que à SC. A tostagem dos grãos de soja a 145°C(STC1)contribuiu para uma menor degradabilidade ruminal da proteína bruta.

RESUMO

Objetivou-se estudar o efeito de diferentes tratamentos térmicos, tempos e procedimentos na degradação ruminal de grãos de soja crus e tostados e sua ação na digestão intestinal da proteína não degradada no rúmen (PNDR) pelo método dos três estágios. Para a degradação ruminal in situ foram pesados cinco gramas de matéria natural em sacos de náilon incubados durante 2; 4; 8; 16; 24 e 48 horas. No tempo zero foi efetuado o mesmo procedimento, excetuando a incubação ruminal. Os resíduos de cada tratamento formaram uma amostra composta para determinar a matéria seca (MS) e proteína bruta (PB). Para a digestibilidade intestinal realizou-se a incubação in situ por 16 horas, usando a técnica dos três estágios. A degradabilidade efetiva da MS com taxa de passagem de 5%/hora para a soja crua (SC) foi de 71,94% e tostada entre 52,23% a 68,78%. Após 16 horas de incubação a PNDR variou de 32,12 a 67,72% e a digestibilidade intestinal de 73,21% a 86,02%. A menor degradação da MS e PB foi da soja tostada a 145oC durante um minuto com steeping (STC1). A digestibilidade intestinal in vitro dos grãos crus foi superior e diferiu dos tostados, exceto a soja tostada a 115oC durante quatro minutos com steeping. A menor degradação proteica foi obtida da STC1de 67,72% da PNDR,52,33% a mais do que à SC. A tostagem dos grãos de soja a 145oC(STC1)contribuiu para uma menor degradabilidade ruminal(AU)

This study analyse ruminal degradation the technique of the nylon bags of dry matter ( DM) and crude protein (CP ) and the intestinal digestibility of rumen undegraded protein (RUP) by the method of the three stages of raw and roasted soybeans at different temperatures with and without The in situ ruminal degradation were weighed five grams of natural matter in nylon bags incubated 2; 4;8; 16; 24 and 48 hours. Zero time was made the same procedure, except ruminal incubation. The residue of treatment formed a composite sample to determine the DM and CP. The intestinal digestibility was in situ incubation for 16 hours, using the technique of the three stages. The degradability of DM at a passage rate of 5 % / hour, for raw soybean (RS), was 71.94 % and roasted between 52.23 % and 68.78 %. After 16 hours of incubation(RUP) ranged from 32.12 to 67.72% and the intestinal digestibility of 73.21% to 86.02 %. The lowest degradation of DM and CP was in the roasted soybeans at 145ºC for one minute with steeping (STC1). The in vitro intestinal digestibility of raw grains was higher and differed from the toasted, except for the ones toasted at 115 ºC for four minutes with steeping. For STC1, was obtained the lowest protein degradation of 67.72 % RUP, which 52.33 %, more when compare to RS. The toast of soybeans at 145 ºC (STC1) contributed to a lower ruminal degradability of crude protein.(AU)

Assuntos

RESUMO

Free films were obtained by the solvent casting method from retrograded starch-pectin dispersions at different polymer proportions and concentrations with and without plasticizer. Film forming dispersions were characterized according to their hardness, birefringence and rheological properties. The polymer dispersions showed a predominantly viscous behavior (G″>G') and the absence of plasticizers lead to building of stronger structures, while the occurrence of Maltese crosses in the retrograded dispersions indicates the occurrence of a crystalline organization. Analyses of the films included mechanical properties, thickness, superficial and cross sectional morphology, water vapor permeability, liquid uptake ability, X-ray diffractometry, in vitro dissolution and enzymatic digestion. The high resistant starch content (65.8-96.8%) assured the resistance of materials against enzymatic digestion by pancreatin. Changes in the X-ray diffraction patterns indicated a more organized and crystalline structure of free films in relation to isolated polymers. Increasing of pectin proportion and pH values favored the dissolution and liquid uptake of films. Films prepared with lower polymer concentration presented better barrier function (WVP and mechanical properties).

Assuntos

RESUMO

A obtenção de protoplastos de fungos, utilizando-se enzimas degradadoras de parede celular, tem sido o método mais utilizado em processos de transformação genética. Foram testados dois tipos de estruturas fúngicas (micélio e conídios), diferentes concentrações enzimáticas (5, 10, 20 mg), estabilizadores osmóticos (NaCl 0,7 mol.L-1 pH 5,7; (NH4)2SO4 1,2 mol.L-1 pH 5,8; KCl 0,7 mol.L-1 pH 5,8; MgSO4 0,7 mol.L-1 pH 5,5; Sacarose 0,5 mol.L-1 pH 5,7; SorbitoL 0,6 mol.L-1 pH 5,7) e seis tempos de exposição dos protoplastos ao sistema lítico, para estabelecer condições otimizadas de obtenção e regeneração de protoplastos de Colletotrichum gloeosporioides, agente relacionado a mancha manteigosa em cafeeiros. Protoplastos de C. gloeosporiodes foram obtidos em maior quantidade quando o micélio foi exposto durante 4 horas, com 10 mg.mL-1 de Lysing Enzime em KCl 0,7 mol.L-1 que se apresentou como melhor estabilizador osmótico, com frequência de regeneração de 11,64%.

The isolation and regeneration of protoplasts from fungal cells, using several cell wall degrading enzymes, has been the most common method to prepare competent cells for genetic studies of filamentous fungi. In this work two types of fungal structures, different enzyme concentrations (5, 10, 20 mg), osmotic stabilizers (NaCl 0,7 mol.L-1 pH 5,7; (NH4)2SO4 1,2 mol.L-1 pH 5,8; KCl 0,7 mol.L-1 pH 5,8; MgSO4 0,7 mol.L-1 pH 5,5; Sacarose 0,5 mol.L-1 pH 5,7; Sorbitol 0,6 mol.L-1 pH 5,7), and six exposure times to establish optimum conditions of isolation and regeneration of protoplasts of Colletotrichum gloeosporioides, agent of blister spot on coffee, were tested. Protoplast of C. gloeosporioides were obtained in greater quantities when the mycelium was exposed for 5 hours with 15mg.mL-1 of Lysing Enzyme in 0.7 mol.L -1 of KCl as osmotic stabilizer, presenting a regeneration rate of 11.64%.

Assuntos