Effects of hot and cold smoking processes on organoleptic properties, yield and composition of matrinxa fillet

Foi avaliado o efeito do processo de defumação a quente (45-90ºC/5 horas) e a frio (27-45ºC/10 horas) nas propriedades organolépticas, no rendimento e na composição dos filés de matrinxã (Brycon cephalus). Não houve diferença significativa no rendimento de filés defumados e não-defumados. As perdas no processo de defumação foram significativamente maiores para defumação a quente (19,37%) em comparação à defumação a frio (17,08%). O processo de defumação reduziu a umidade (in natura = 72,91%; defumado a quente = 58,51%; e defumado a frio = 59,68%) e aumentou os teores de proteína bruta, lipídios e cinzas. Houve diferença significativa somente nos teores de proteína no defumado a quente (28,07%) e defumado a frio (27,14%). O processo a frio resultou em melhor aparência e cor de filé, enquanto o processo a quente melhorou o sabor, o teor de sal e a aparência geral. O aroma e a textura não diferiram significativamente entre os processos. O processo de defumação a quente melhora as propriedades organolépticas e os níveis de proteína do filé de matrinxã.

Desempenho de juvenis de tilápia-do-nilo alimentados com rações contendo complexo enzimático

Avaliou-se o efeito da inclusão de um complexo enzimático em dietas para tilápias-do-nilo (Oreochromis niloticus) sobre o desempenho, a composição química da carcaça e a qualidade da água. Foram utilizados 200 alevinos revertidos (4,57 ± 1,24 g), distribuídos em delineamento inteiramente casualizado em 20 tanques de 500 litros, com quatro tratamentos e cinco repetições, considerando a unidade experimental uma caixa com dez peixes. Os peixes foram alimentados com dietas contendo 0; 0,033; 0,066 ou 0,099% de complexo enzimático. As dietas foram processadas na forma peletizada e fornecidas quatro vezes ao dia, às 8, 11, 14 e 17 h. Os valores médios de pH, condutividade elétrica, oxigênio dissolvido, temperatura, fósforo total, amônia e nitrato da água de cultivo não foram influenciados pela dieta. A inclusão do complexo enzimático na dieta não afetou o ganho de peso, as taxas de sobrevivência e de crescimento específico, mas influenciou o consumo de ração e a conversão alimentar, cujos valores foram maiores nos peixes alimentados com a dieta com 0,066% de complexo enzimático. Não foram observadas diferenças nos teores de matéria seca, umidade, proteína bruta, matéria mineral, cálcio e fósforo na carcaça dos peixes, no entanto, o teor de extrato etéreo reduziu de forma linear com o aumento do nível de complexo enzimático. A utilização de complexo enzimático (amilase, protease, celulase, lipase, pectinase, xilanase, β-glucanase e fitase) no nível de 0,066% em dietas para juvenis de tilápia-do-nilo piora a conversão alimentar, mas não influencia o desempenho e a composição corporal dos peixes.

COMPOSITION OF FRESHLY HARVESTED BRAZILIAN ROYAL JELLY - IDENTIFICATION OF CARBOHYDRATES FROM THE SUGAR FRACTION

Samples of Brazilian royal jelly from Africanized Apis mellifera were analysed in order to determine the gross composition: crude moisture ranged from 67.80% to 69.40%, crude protein from 15.80% to 16.70%, crude lipid from 2.90% to 3.98% and-total sugars from 11.40% to 11.50%. The sugar fraction was investigated and revealed the presence of the following compounds identified by their retention time during HPLC analysis: ribose, fructose, glucose, sucrose, mannose, trehalose, erythritol, adonitol and mannitol.

Evaluation of physicochemical and glycaemic properties of commercial plant-based milk substitutes

The market for plant-based dairy-type products is growing as consumers replace bovine milk in their diet, for medical reasons or as a lifestyle choice. A screening of 17 different commercial plant-based milk substitutes based on different cereals, nuts and legumes was performed, including the evaluation of physicochemical and glycaemic properties. Half of the analysed samples had low or no protein contents (<0.5 %). Only samples based on soya showed considerable high protein contents, matching the value of cow’s milk (3.7 %). An in-vitro method was used to predict the glycaemic index. In general, the glycaemic index values ranged from 47 for bovine milk to 64 (almond-based) and up to 100 for rice-based samples. Most of the plant-based milk substitutes were highly unstable with separation rates up to 54.39 %/h. This study demonstrated that nutritional and physicochemical properties of plant-based milk substitutes are strongly dependent on the plant source, processing and fortification. Most products showed low nutritional qualities. Therefore, consumer awareness is important when plant-based milk substitutes are used as an alternative to cow’s milk in the diet.

Spatio-temporal patterns of herbage availability and livestock movements: A cross-border analysis in the Chinese-Mongolian Altay

Due to increasing population and the recent implementation of policies to intensify the use of land and water resources, the transhumant pastoral systems in the Chinese-Mongolian Altay-Dzungarian region are rapidly changing, leading to modifications of herd size, herd composition and spatial distribution of livestock grazing. This may have major consequences for the supply and quality of rangeland biomass. Despite similar topographic settings, the socio-political framework for Chinese and Mongolian pastoralists differs significantly, leading to differences in rangeland utilization. To substantiate these claims, the long-distance transhumance routes, frequency of pasture changes, daily grazing itineraries and size of pastures were recorded by means of GPS tracking of cattle and goats on 1,535 (China) and 1,396 (Mongolia) observation days. The status quo of the main seasonal pastures was captured by measuring the herbage offer and its nutritive value in 869 sampling spots. In the Altay-Dzungarian region, small ruminant herds covered up to 412 km (Mongolia) and grazed on up to nine pastures per year (China). In Mongolia, the herds’ average duration of stay at an individual pasture was longer than in China, particularly in spring and autumn. Herbage allowance at the onset of a grazing period (kg dry matter per sheep unit and day) ranged from 34/17 to 91/95 (China/Mongolia). Comparing crude protein and phosphorous concentrations of herbage, in China, the highest concentrations were measured for spring and summer pastures, whereas in Mongolia, the highest concentrations were determined for autumn and winter pastures. Based on our data, we conclude that regulation of animal numbers and access to pastures seemingly maintained pasture productivity in China, especially at high altitudes. However, this policy may prohibit flexible adaptation to sudden environmental constraints. In contrast, high stocking densities and grazing of pastures before flowering of herbaceous plants negatively affected rangeland productivity in Mongolia, especially for spring and summer pastures.

Annual crop rotation of tropical pastures with no-till soil as affectedby lime surface application.

Soil acidity and low natural fertility are the main limiting factors for grain production in tropical regionssuch as the Brazilian Cerrado. The application of lime to the surface of no-till soil can improve plant nutrition, dry matter production, crop yields and revenue. The present study, conducted at the Lageado Experimental Farm in Botucatu, State of São Paulo, Brazil, is part of an ongoing research project initi-ated in 2002 to evaluate the long-term effects of the surface application of lime on the soil?s chemical attributes, nutrition and kernel/grain yield of peanut (Arachis hypogaea), white oat (Avena sativa L.) and maize (Zea mays L.) inter cropped with palisade grass (Urochloa brizantha cv. Marandu), as well as the forage dry matter yield of palisade grass in winter/spring, its crude protein concentration, estimated meat production, and revenue in a tropical region with a dry winter during four growing seasons. The experiment was designed in randomized blocks with four replications. The treatments consisted of four rates of lime application (0, 1000, 2000 and 4000 kg ha−1), performed in November 2004. The surface application of limestone to the studied tropical no-till soil was efficient in reducing soil acidity from the surface down to a depth of 0.60 m and resulted in greater availability of P and K at the soil surface. Ca and Mg availability in the soil also increased with the lime application rate, up to a depth of 0.60 m. Nutrient absorption was enhanced with liming, especially regarding the nutrient uptake of K, Ca and Mg by plants.Significant increases in the yield components and kernel/grain yields of peanut, white oat and maize were obtained through the surface application of limestone. The lime rates estimated to achieve the maximum grain yield, especially in white oat and maize, were very close to the rates necessary to increase the base saturation of a soil sample collected at a depth of 0?0.20 m to 70%, indicating that the surface liming of 2000 kg ha−1is effective for the studied tropical no-till soil. This lime rate also increases the forage dry matter yield, crude protein concentration and estimated meat production during winter/spring in the maize-palisade grass inter cropping, provides the highest total and mean net profit during the four growing seasons, and can improve the long-term sustainability of tropical agriculture in the Brazilian Cerrado.

Composição botânica e qualidade do pasto selecionado por ovelhas em caatinga raleada e enriquecida.

Resumo: O presente estudo foi conduzido com o objetivo de determinar a composição botânica e a qualidade da dieta selecionada por ovelhas, através da técnica de micro-histologia fecal, em caatinga raleada e enriquecida com capim massai (Panicum maximum cv. Massai), recebendo diferentes quantidades de concentrado (0; 200; 350 e 500 g de concentrado por dia), e em diferentes períodos do ano (águas, transição água-seca e seca). Foram estimados também o consumo e digestibilidade dos nutrientes, bem como a degradabilidade de espécies forrageiras ingeridas pelas ovelhas. Os experimentos foram realizados na Fazenda Crioula do Meio, pertencente a Embrapa Caprinos e Ovinos em Sobral, CE no período de março a novembro de 2013. No Experimento 1, para a determinação da composição botânica e qualidade da dieta selecionada, foram utilizadas dezesseis ovelhas Somalis brasileira, gestantes, multíparas e peso médio de 30,58+2,48 kg. O acompanhamento da ingestão do pasto pelas ovelhas foi feito em três períodos (águas, transição água-seca e seca), referentes aos meses de abril, junho e agosto de 2013, respectivamente. Amostras das plantas foram coletadas para o preparo das lâminas de referência, e posterior identificação e caracterização dos descritores epidérmicos. O mesmo foi feito para as fezes coletadas nas ovelhas. Com base na proporção de cada espécie identificada nas lâminas fecais que compuseram a dieta, e na composição química das forrageiras identificadas, foi possível determinar a qualidade da dieta ingerida. De 76 espécies observadas no pasto, 33 foram identificadas na dieta das ovelhas, destacando as espécies sabiá (Mimosa caesalpiniaefolia), centrosema (Centrosema sp.), ervanço (Alternanthera brasiliana), massai (Panicum maximum cv Massai) e paco-paco (Wissadula rostrata) como as mais selecionadas pelos animais ao longo dos períodos, chegando a compor mais de 50% da dieta selecionada. Com a chegada do período seco, espécies indesejáveis como o marmeleiro (Croton sonderianus) e o mofumbo (Combretum lepreosum), também fizeram parte das plantas selecionadas. Quanto ao valor nutritivo da dieta selecionada, os animais selecionaram uma dieta com valor nutritivo superior ao amostrado no pasto. No Experimento 2, na mesma condição do experimento anterior, trinta e duas ovelhas Somalis brasileira foram utilizadas para determinação do consumo e digestibilidade dos nutrientes, realizado em três ensaios (abril - terço final de gestação; junho - lactação e agosto - desmame). Para predição do consumo, o indicador LIPE® foi utilizado. Pesagens quinzenais foram realizadas para avaliação do desempenho das ovelhas e dos cordeiros nascidos. O concentrado oferecido favoreceu a maior ingestão e digestibilidade da MS e PB, com efeito substitutivo em relação ao consumo de pasto (P<0,05). Para o período seco, menores consumos foram observados em relação aos períodos das águas e de transição água-seca (P<0,05). Maiores consumos e digestibilidades dos constituintes fibrosos foram observados para as ovelhas não suplementados (P<0,05). Na avaliação do desempenho, a suplementação oferecida determinou os maiores pesos verificados durante a lactação, ao desmame e para os pesos ao nascer e ao desmame dos cordeiros (P<0,05). No Experimento 3, dois ovinos Morada Nova foram utilizados para determinação da degradabilidade da matéria seca (MS), proteína bruta (PB) e fibra em detergente neutro (FDN) de cinco das forrageiras selecionadas pelas ovelhas no Experimento 1: M. caesalpiniaefolia, A. brasiliana, P. maximum cv. Massai, jurema-preta (Mimosa tenuiflora), C. leprosum, nos tempos 0, 6, 24, 48, 72 e 96 horas de incubação. Para cada forrageira, foram determinadas equações para o desaparecimento da MS, PB e FDN. Também foi feito o fracionamento da proteína em suas porções degradáveis e não degradáveis no rúmen. Foi observado maior desaparecimento da MS, PB e FDN, além dos melhores níveis de proteína efetivamente degradada no rúmen para A. brasiliana, seguido pelo P. maximum cv. Massai e M. caesalpiniaefolia. Com as informações obtidas, conclui-se que a micro-histologia fecal apresenta-se como uma técnica viável para avaliações da composição botânica da dieta selecionada por ovinos na caatinga. Ovelhas na caatinga possuem uma grande habilidade de selecionar a dieta, modificando-a ao longo das fases fenológicas, sempre na tentativa de estabelecer uma dieta com melhor valor nutritivo. Forrageiras como A. brasiliana, M. caesalpiniaefolia e o P. maximum cv. Massai, podem ser consideradas um interessante recurso alimentar, em virtude de seu valor nutricional e aproveitamento por ovelhas criadas na caatinga. Abstract: This study was conducted in order to determine the botanical composition and diet quality selected by sheep through fecal micro-histological technique, in thinned and enriched caatinga with Massai grass (Panicum maximum cv Massai.), receiving different amounts of concentrate (0; 200; 350 and 500 g of concentrate per day) at different periods (wet, transition wet-dry and dry). Were also estimated the intake, digestibility, as well as the degradability of forage species eaten by sheep. The experiments were performed in the "Fazenda Crioula do Meio", owned by Embrapa Goats and Sheep, in Sobral, Ceará State, Brazil, from march to november 2013. In Trial 1, sixteen female, pregnant, multiparous, with average body weight of 30,58+2,48 kg Somalis brasileira breed sheep were used to determine the botanical composition and the quality of the selected diet. The monitoring of pasture intake of sheep were conducted in three phenological periods of the caatinga's pasture (wet season, transition wet-dry and dry season). Plant samples were collected for the preparation of the reference slides, with subsequent identification and characterization of epidermal descriptors. The same was done for the feces collected in sheep. Considering the proportion of each species identified in fecal slides which composed the diet, and the chemical composition of forage identified, it was possible to determine the quality of the selected diet. From 76 species observed in the pasture, 33 species was identified in the sheep selected diet, emphasizing the Sabiá (Mimosa caesalpiniaefolia), centrosema (Centrosema sp.), ervanço (Alternanthera brasiliana), massai (Panicum maximum cv Massai) e paco-paco (Wissadula rostrata) as the most selected species by sheep during the study, composing more than 50% of the selected diet. In the Dry Season, undesirable species like marmeleiro (Croton sonderianus) and mofumbo (Combretum leprosum), were also constituent of the diet. Regarding to the nutritional value of selected diet, the sheep selected a diet with more protein than the sampled in the pasture. In the final late gestation, the sheep without supplementation ate a diet above 16% of CP, higher than the selected diet by treatments 350 and 500 g of concentrate per day (P<0.05). Were also observed to the non supplemented sheep, less fiber content intake (P<0.05). In the Trial 2, in the same condition of the first Trial, thirty two Somalis brasileira female sheep were used to determine the intake and digestibility, conducted in three assays (April - third late pregnancy; June - lactation and August - weaning). To predict the intake, the marker LIPE was used. Sheep and lambs were weighted every two weeks to performance evaluation. The concentrate offered to sheep favored to higher intake and digestibility of DM and CP than non supplemented sheep, with inverse relationship to the pasture intake (P<0.05). For the Dry Season, lower intake were observed than for Wet Season and Transition Wet-Dry (P<0.05). Higher intakes and digestibility of the fiber constituents were verified to non supplemented sheep (P<0.05). To performance evaluation, the offered supplementation determined the higher weights observed during lactation and weaning of sheep, and to birth and weaning weights of lambs (P<0.05). In Trial 3, two male sheep were used to determine the degradability of dry matter (DM), crude protein (CP) and neutral detergent fiber (NDF) of five forages selected by sheep in Trial 1: M. caesalpiniaefolia, A. brasiliana, P. maximum cv. Massai, jurema-preta (Mimosa tenuiflora) and C. lepreosum, at zero, 6, 24, 48, 72 and 96 hours of incubation. For each forage, were determined equations for the disappearance of DM, CP and NDF. It was also realized the protein fractions in their degradable and non-degradable in the rumen parts. Was detected a higher disappearance of DM, CP and NDF, as also better proportion of the rumen degradable protein to A. brasiliana, followed by P. maximum cv. Massai and M. caesalpiniaefolia. With the information obtained, it is concluded that the fecal micro histological technique presents as a viable technique to evaluate the selected diet by sheep in caatinga's pasture. On this pasture, the sheep are skilled to select the diet, changing during the phenological phases, trying to form a diet of better nutritive value. Forages as A. brasiliana, P. maximum cv. Massai and M. caesalpiniaefolia, can be considered an interesting food source to ewes kept in the caatinga.

Forage peanut (Arachis spp.) genetic evaluation and selection.

Forage peanut improvement for use in grass?legume mixtures is expected to have a great impact on the sustainability of Brazilian livestock production. Eighteen cloned Arachis spp. ecotypes were evaluated under clipping in a Brazilian Cerrado region and results analysed using a mixed model methodology. The objective was to estimate genetic and phenotypic parameters and to select the best ecotypes based on selection index applied on their predicted genotypic value. The traits of total dry-matter (DM) and leaf DM yield presented moderate (0_30 < h2g < 0_50) to high (>0_50) broad-sense heritability, in contrast to the low genetic variability in nutritional quality-associated traits. Ecotypes of Arachis spp. contained average crude protein concentrations of 224 g kg _1 DM in leaves and 138 g kg _1 DM in stems, supporting the potential role of these species to overcome the low protein content in Cerrado pastures. The correlations between yield traits and traits associated with low nutritional value in leaves were consistently significant and positive. Genetic correlations among all the yield traits evaluated during the rainy or dry seasons were significant and positive. The ecotypes were ranked based on selection index. The next step is to validate long-term selection of grass?Arachis in combination with pastures under competition and adjusted grazing in the Cerrado region.

Uso da espectroscopia de reflectância do infravermelho próximo (NIRS) para previsão da composição bromatológica de vagens de algaroba e palma forrageira.

Espécies forrageiras adaptadas às condições semiáridas são uma alternativa para reduzir os impactos negativos na cadeia produtiva de ruminantes da região Nordeste brasileira devido à sazonalidade na oferta de forragem, além de reduzir custo com o fornecimento de alimentos concentrados. Dentre as espécies, a vagem de algaroba (Prosopis juliflora SW D.C.) e palma forrageira (Opuntia e Nopalea) ganham destaque por tolerarem o déficit hídrico e produzirem em períodos onde a oferta de forragem está reduzida, além de apresentam bom valor nutricional e serem bem aceitas pelos animais. Porém, devido à variação na sua composição, seu uso na alimentação animal exige o conhecimento profundo da sua composição para a elaboração de dietas balanceadas. No entanto, devido ao custo e tempo para análise, os produtores não fazem uso da prática de análise da composição químico-bromatológica dos alimentos. Por isto, a espectroscopia de reflectância no infravermelho próximo (NIRS) representa uma importante alternativa aos métodos tradicionais. Objetivou-se com este estudo desenvolver e validar modelos de predição da composição bromatológica de vagem de algaroba e palma forrageira baseados em espectroscopia NIRS, escaneadas em dois modelos de equipamentos e com diferentes processamentos da amostra. Foram coletadas amostras de vagem de algaroba nos estados do Ceará, Bahia, Paraíba e Pernambuco, e amostras de palma forrageira nos estados do Ceará, Paraíba e Pernambuco, frescas (in natura) ou pré-secas e moídas. Para obtenção dos espectros utilizaram-se dois equipamentos NIR, Perten DA 7250 e FOSS 5000. Inicialmente os alimentos foram escaneados in natura em aparelho do modelo Perten, e, com o auxílio do software The Unscrambler 10.2 foi selecionado um grupo de amostras para o banco de calibração. As amostras selecionadas foram secas e moídas, e escaneadas novamente em equipamentos Perten e FOSS. Os valores dos parâmetros de referência foram obtidos por meio de metodologias tradicionalmente aplicadas em laboratório de nutrição animal para matéria seca (MS), matéria mineral (MM), matéria orgânica (MO), proteína bruta (PB), estrato etéreo (EE), fibra solúvel em detergente neutro (FDN), fibra solúvel em detergente ácido (FDA), hemicelulose (HEM) e digestibilidade in vitro da matéria seca (DIVMS). O desempenho dos modelos foi avaliado de acordo com os erros médios de calibração (RMSEC) e validação (RMSECV), coeficiente de determinação (R2 ) e da relação de desempenho de desvio dos modelos (RPD). A análise exploratória dos dados, por meio de tratamentos espectrais e análise de componentes principais (PCA), demonstraram que os bancos de dados eram similares entre si, dando segurança de desenvolver os modelos com todas as amostras selecionadas em um único modelo para cada alimento, algaroba e palma. Na avaliação dos resultados de referência, observou-se que a variação dos resultados para cada parâmetro corroboraram com os descritos na literatura. No desempenho dos modelos, aqueles desenvolvidos com pré-processamento da amostra (pré-secagem e moagem) se mostraram mais robustos do que aqueles construídos com amostras in natura. O aparelho NIRS Perten apresentou desempenho semelhante ao equipamento FOSS, apesar desse último cobrir uma faixa espectral maior e com intervalos de leituras menores. A técnica NIR, associada ao método de calibração multivariada de regressão por meio de quadrados mínimos (PLS), mostrou-se confiável para prever a composição químico-bromatológica de vagem de algaroba e da palma forrageira. Abstract: Forage species adapted to semi-arid conditions are an alternative to reduce the negative impacts in the feed supply for ruminants in the Brazilian Northeast region, due to seasonality in forage availability, as well as in the reducing of cost by providing concentrated feedstuffs. Among the species, mesquite pods (Prosopis juliflora SW DC) and spineless cactus (Opuntia and Nopalea) are highlighted for tolerating the drought and producion in periods where the forage is scarce, and have high nutritional value and also are well accepted by the animals. However, its use in animal diets requires a knowledge about its composition to prepare balanced diets. However, farmers usually do not use feed composition analysis, because their high cost and time-consuming. Thus, the Near Infrared Reflectance Spectroscopy in the (NIRS) is an important alternative to traditional methods. The objective of this study to develop and validate predictive models of the chemical composition of mesquite pods and spineless cactus-based NIRS spectroscopy, scanned in two different spectrometers and sample processing. Mesquite pods samples were collected in the states of Ceará, Bahia, Paraiba and Pernambuco, and samples of forage cactus in the states of Ceará, Paraíba and Pernambuco. In order to obtain the spectra, it was used two NIR equipment: Perten DA 7250 and FOSS 5000. sSpectra of samples were initially obtained fresh (as received) using Perten instrument, and with The Unscrambler software 10.2, a group of subsamples was selected to model development, keeping out redundant ones. The selected samples were dried and ground, and scanned again in both Perten and FOSS instruments. The values of the reference analysis were obtained by methods traditionally applied in animal nutrition laboratory to dry matter (DM), mineral matter (MM), organic matter (OM), crude protein (CP), ether extract (EE), soluble neutral detergent fiber (NDF), soluble acid detergent fiber (ADF), hemicellulose ( HEM) and in vitro digestibility of dry matter (DIVDM). The performance of the models was evaluated according to the Root Mean Square Error of Calibration (RMSEC) and cross-validation (RMSECV), coefficient of determination (R2 ) and the deviation of Ratio of performance Deviation of the models (RPD). Exploratory data analysis through spectral treatments and principal component analysis (PCA), showed that the databases were similar to each other, and may be treated asa single model for each feed - mesquite pods and cactus. Evaluating the reference results, it was observed that the variation were similar to those reported in the literature. Comparing the preprocessing of samples, the performance ofthose developed with preprocessing (dried and ground) of the sample were more robust than those built with fresh samples. The NIRS Perten device performance similar to FOSS equipment, although the latter cover a larger spectral range and with lower readings intervals. NIR technology associate do multivariate techniques is reliable to predict the bromatological composition of mesquite pods and cactus.

Substitution of crude cell wall for neutral detergent fibre in the equations of the Cornell Net Carbohydrate and Protein System that predict carbohydrate fractions: application to sunflower (Helianthus annulus L.)

Prediction of carbohydrate fractions using equations from the Cornell Net Carbohydrate and Protein System (CNCPS) is a valuable tool to assess the nutritional value of forages. In this paper these carbohydrate fractions were predicted using data from three sunflower (Helianthus annuus L.) cultivars, fresh or as silage. The CNCPS equations for fractions B(2) and C include measurement of ash and protein-free neutral detergent fibre (NDF) as one of their components. However, NDF lacks pectin and other non-starch polysaccharides that are found in the cell wall (CW) matrix, so this work compared the use of a crude CW preparation instead of NDF in the CNCPS equations. There were no differences in the estimates of fractions B, and C when CW replaced NDF; however there were differences in fractions A and B2. Some of the CNCPS equations could be simplified when using CW instead of NDF Notably, lignin could be expressed as a proportion of DM, rather than on the basis of ash and protein-free NDF, when predicting CNCPS fraction C. The CNCPS fraction B(1) (starch + pectin) values were lower than pectin determined through wet chemistty. This finding, along with the results obtained by the substitution of CW for NDF in the CNCPS equations, suggests that pectin was not part of fraction B(1) but present in fraction A. We suggest that pectin and other non-starch polysaccharides that are dissolved by the neutral detergent solution be allocated to a specific fraction (B2) and that another fraction (B(3)) be adopted for the digestible cell wall carbohydrates.

Adjuvant requirement for successful immunization with recombinant derivatives of Plasmodium vivax merozoite surface protein-1 delivered via the intranasal route

Recently, we generated two bacterial recombinant proteins expressing 89 amino acids of the C-terminal domain of the Plasmodium vivax merozoite surface protein-1 and the hexa-histidine tag (His6MSP1(19)). One of these recombinant proteins contained also the amino acid sequence of the universal pan allelic T-cell epitope (His6MSP1(19)-PADRE). In the present study, we evaluated the immunogenic properties of these antigens when administered via the intra-nasal route in the presence of distinct adjuvant formulations. We found that C57BL/6 mice immunized with either recombinant proteins in the presence of the adjuvants cholera toxin (CT) or the Escherichia coli heat labile toxin (LT) developed high and long lasting titers of specific serum antibodies. The induced immune responses reached maximum levels after three immunizing doses with a prevailing IgG1 subclass response. In contrast, mice immunized by intranasal route with His6MSP1(19)-PADRE in the presence of the synthetic oligonucleotides adjuvant CpG ODN 1826 developed lower antibody titers but when combined to CT, CpG addition resulted in enhanced IgG responses characterized by lower IgG1 levels. Considering the limitations of antigens formulations that can be used in humans, mucosal adjuvants can be a reliable alternative for the development of new strategies of immunization using recombinant proteins of P. vivax.

Requirement of mitogen-activated protein kinase kinase 3 (MKK3) for tumor necrosis factor-induced cytokine expression

The p38 mitogen-activated protein kinase is activated by treatment of cells with cytokines and by exposure to environmental stress. The effects of these stimuli on p38 MAP kinase are mediated by the MAP kinase kinases (MKKs) MKK3, MKK4, and MKK6. We have examined the function of the p38 MAP kinase signaling pathway by investigating the effect of targeted disruption of the Mkk3 gene. Here we report that Mkk3 gene disruption caused a selective defect in the response of fibroblasts to the proinflammatory cytokine tumor necrosis factor, including reduced p38 MAP kinase activation and cytokine expression. These data demonstrate that the MKK3 protein kinase is a critical component of a tumor necrosis factor-stimulated signaling pathway that causes increased expression of inflammatory cytokines.

Multiple pathways to bypass the enhancer requirement of sigma 54 RNA polymerase: Roles for DNA and protein determinants

Sigma 54 is a required factor for bacterial RNA polymerase to respond to enhancers and directs a mechanism that is a hybrid between bacterial and eukaryotic transcription. Three pathways were found that bypass the enhancer requirement in vitro. These rely on either deletion of the sigma 54 N terminus or destruction of the DNA consensus −12 promoter recognition element or altering solution conditions to favor transient DNA melting. Each of these allows unstable heparin-sensitive pre-initiation complexes to form that can be driven to transcribe in the absence of both enhancer protein and ATP β–γ hydrolysis. These disparate pathways are proposed to have a common basis in that multiple N-terminal contacts may mediate the interactions between the polymerase and the DNA region where melting originates. The results raise possibilities for common features of open complex formation by different RNA polymerases.

Assembly of a Novel Cartilage Matrix Protein Filamentous Network: Molecular Basis of Differential Requirement of von Willebrand Factor A Domains

Cartilage matrix protein (CMP) is the prototype of the newly discovered matrilin family, all of which contain von Willebrand factor A domains. Although the function of matrilins remain unclear, we have shown that, in primary chondrocyte cultures, CMP (matrilin-1) forms a filamentous network, which is made up of two types of filaments, a collagen-dependent one and a collagen-independent one. In this study, we demonstrate that the collagen-independent CMP filaments are enriched in pericellular compartments, extending directly from chondrocyte membranes. Their morphology can be distinguished from that of collagen filaments by immunogold electron microscopy, and mimicked by that of self-assembled purified CMP. The assembly of CMP filaments can occur from transfection of a wild-type CMP transgene alone in skin fibroblasts, which do not produce endogenous CMP. Conversely, assembly of endogenous CMP filaments by chondrocytes can be inhibited specifically by dominant negative CMP transgenes. The two A domains within CMP serve essential but different functions during network formation. Deletion of the A2 domain converts the trimeric CMP into a mixture of monomers, dimers, and trimers, whereas deletion of the A1 domain does not affect the trimeric configuration. This suggests that the A2 domain modulates multimerization of CMP. Absence of either A domain from CMP abolishes its ability to form collagen-independent filaments. In particular, Asp22 in A1 and Asp255 in A2 are essential; double point mutation of these residues disrupts CMP network formation. These residues are part of the metal ion–dependent adhesion sites, thus a metal ion–dependent adhesion site–mediated adhesion mechanism may be applicable to matrilin assembly. Taken together, our data suggest that CMP is a bridging molecule that connects matrix components in cartilage to form an integrated matrix network.