ABSTRACT
Bioactive fatty acids present in goat milk have the ability to reduce the risks of coronary heart disease in humans, and condensed tannins (CT) can modulate the polyunsaturated fatty acids (PUFA) biohydrogenation process in the rumen and consequently increase the levels of these fatty acids. Thus, the objective was to evaluate the inclusion of CT in the diet for Canindé, Repartida, and Saanen goats to increase the level of bioactive fatty acids in milk. Twenty-two lactating does of three genetic groups, six Canindé, eight Repartida, and eight Saanen, were randomly assigned in a 3 × 2 factorial arrangement of three genetic groups and two diets (control and with 50 g CT/kg DM). The inclusion of CT in the diet did not change (P > 0.05) nutrient intake and performance. However, the inclusion of CT promoted an increase (P < 0.05) in C14:1; cis-9; C18:2n6; C18:3n6; C18:3n3; PUFA; and long-chain fatty acids and reduction (P < 0.05) of C11; C12; C14; ω6/ω3; and atherogenicity index in milk fat. Thus, it is recommended to include CT in the diet for Canindé, Repartida, and Saanen goats to increase the level of bioactive fatty acids in milk. The inclusion of the tannins of Acacia mearnsii promoted an increase in C14:1; cis-9; C18:2n6; C18:3n6; C18:3n3; polyunsaturated fatty acid; and long-chain fatty acids and reduction of C11; C12; C14; ω6/ω3; and atherogenicity index in milk fat.
Subject(s)
Milk , Proanthocyanidins , Animals , Female , Animal Feed/analysis , Diet/veterinary , Fatty Acids , Goats , Lactation , RumenABSTRACT
Este artigo tem como objetivo discutir algumas das maneiras como o conceito heideggeriano de "ser para a morte" é decisivo para a tarefa da ontologia fundamental delineada por Heidegger em Ser e tempo. Partindo da assunção de que o tempo é o horizonte da compreensão do ser em geral, o modo originário de doação do ser deverá ser compreendido temporalmente. À medida que a temporalidade do ser-aí é primeiro desentranhada por meio da análise do ser para a morte como sua possibilidade mais própria, não será suficiente o esclarecimento das estruturas da existência na forma de uma estrutura conceitual do tempo; porém, o modo da doação de ser deverá efetivar-se na morte. A tarefa da ontologia fundamental, ou seja, a pergunta por como podemos em geral saber não poderá mais ser resolvida nem na relação do ente finito com o infinito transcendente, nem como manifestação infinita da vontade, mas deverá resolver-se a partir da finitude mais radical.
This paper aims to discuss some of the ways in which the heideggerian concept of "being towards death" is decisive for the task of the fundamental ontology as proposed by Heidegger in Being and Time. From the assumption that time is the horizon of the understanding of Being in general, the original mode of donation of being must be temporally understood. Insofar as the temporality of Dasein is first revealed through the analysis of being towards death as its ownmost possibility, it should not be enough to clarify the structures of existence in the form of a conceptual structure of time; the mode of donation of Being should be brought about in death. The task of the fundamental ontology, i. e., the question of how we can in general know, cannot be resolved anymore in the relationship between the finite being and the infinite transcendent, nor as infinite manifestation of will, though it should be resolved from the most radical finitude.
ABSTRACT
A ricina é uma proteína bastante tóxica presente nas sementes de mamona que impossibilita o uso da torta de mamona "in natura", como ração. A torta de mamona destoxificada necessita ainda de métodos de análise que garantam a ausência de traços dessa proteína. Objetivou-se, neste trabalho, produzir e avaliar a sensibilidade e especificidade de anticorpos policlonais anti-ricina, para serem empregados como possíveis componentes de métodos sorológicos na detecção de ricina em torta de mamona destoxificada. Foram avaliadas três doses da proteína: 400, 180 e 100 µg cada uma dividida em duas aplicações em coelhos. A primeira dose foi injetada no animal no início do experimento e a segunda após 21 dias. O método de ELISA indicou que as duas doses menores (100 e 180 µg) induziram respostas imunológicas primária e secundária com produção de anticorpos específicos. Enquanto a dose maior (400 µg) de ricina apresentou uma resposta primária com elevação dos títulos de anticorpos, seguida de uma supressão da resposta. Esse perfil é sugestivo de tolerância imunológica. Pela técnica de Western blotting verificou-se que os anticorpos policlonais produzidos são bastante específicos para a ricina, no entanto, por detectarem ricina na forma nativa e desnaturada não são recomendados para o monitoramento de ricina em torta de mamona destoxificada por tratamento térmico.
Ricin is a very toxic protein found in castor bean plants, making it impossible to use natural castor cake as animal food. The detoxificated castor cake needs to be analyzed by methods that ensure the absence of traces of this protein. This work had the objective to produce and to evaluate the sensitivity and specificity of anti-ricin polyclonal antibodies, to be employed as component of sorologic methods as the ELISA in the detection of ricin in detoxificated castor cake. Three doses of protein, 400, 180 and 100 µg were evaluated each one injected twice into rabbit, with one half in the begin of the experiment and the other half after 21 days of immunization. The ELISA method indicated that the lower doses (100 e 180 µg) induced primary and secondary immunological response with production of specific antibodies, while the higher dose of ricin (400 µg) showed a primary response with increase of the antibody titre, followed of immunological suppression. This profile suggests immunological tolerance. By Western blotting technique it was verified that polyclonal antibodies are too specific to ricin, however, they detected ricin in native and denaturated form and are not recommended for the monitoring of ricin in detoxificated castor bean cake by heat treatment.
ABSTRACT
Frutalin is an alpha-D-galactose-binding lectin expressed in breadfruit seeds. Its isolation from plant is time-consuming and results in a heterogeneous mixture of different lectin isoforms. In order to improve and facilitate the availability of the breadfruit lectin, we cloned an optimised codifying frutalin mature sequence into the pPICZalphaA expression vector. This expression vector, designed for protein expression in the methylotrophic yeast Pichia pastoris, contains the Saccharomyces alpha-factor preprosequence to direct recombinant proteins into the secretory pathway. Soluble recombinant frutalin was detected in the culture supernatants and recognised by native frutalin antibody. Approximately 18-20 mg of recombinant lectin per litre medium was obtained from a typical small scale methanol-induced culture purified by size-exclusion chromatography. SDS-PAGE and Edman degradation analysis revealed that frutalin was expressed as a single chain protein since the four amino-acid linker peptide "T-S-S-N", which connects alpha and beta chains, was not cleaved. In addition, incomplete processing of the signal sequence resulted in recombinant frutalin with one Glu-Ala N-terminal repeat derived from the alpha-factor prosequence. Endoglycosidase treatment and SDS-PAGE analysis revealed that the recombinant frutalin was partly N-glycosylated. Further characterisation of the recombinant lectin revealed that it specifically binds to the monosaccharide Me-alpha-galactose presenting, nevertheless, lesser affinity than the native frutalin. Recombinant frutalin eluted from a size-exclusion chromatography column with a molecular mass of about 62-64 kDa, suggesting a tetrameric structure, however it did not agglutinate rabbit erythrocytes as native frutalin does. This work shows that the galactose-binding jacalin-related lectins four amino-acid linker peptide "T-S-S-N" does not undergo any proteolytic cleavage in the yeast P. pastoris and also that linker cleavage might not be essential for lectin sugar specificity.
