RESUMO
A validated analytical procedure is here described for the quality control of the protein fraction of purified bovine colostrum used in food supplements. The proposed procedure starts with 1D and 2D-gel electrophoresis. The sample is then separated into two fractions by protein G affinity chromatography: the IgG enriched and the IgG depleted fraction (IgG-d). A size exclusion chromatography coupled to UV is then applied to the IgG and IgG-d fractions for the quantitative analysis of IgG and IgM, respectively. The IgG-d fraction is then analysed by HPLC-MS analysis for the quantitative analysis of ß-lactoglobulins and α-lactoalbumin. The next step is to quantitatively measure a set of bioactive proteins selected from the bovine colostrum data bank on the basis of their claimed health benefits. The enzymatic activities of lactoperoxidase and xanthine dehydrogenase/oxidase are then tested as an index of protein functionality.
Assuntos
Colostro/química , Suplementos Nutricionais/análise , Animais , Bovinos , Cromatografia de Afinidade , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Colostro/enzimologia , Colostro/metabolismo , Eletroforese em Gel Bidimensional , Ensaios Enzimáticos , Feminino , Imunoglobulina G/análise , Imunoglobulina M/análise , Lactoperoxidase/análise , Lactoperoxidase/metabolismo , Espectrometria de Massas , Gravidez , Controle de Qualidade , Xantina Desidrogenase/análise , Xantina Desidrogenase/metabolismoRESUMO
Bovine colostrum (BC), the initial milk secreted by the mammary gland immediately after parturition, is widely used for several health applications. We here propose an off-target method based on proteomic analysis to explain at molecular level the potential health benefits of BC. The method is based on the set-up of an exhaustive protein data bank of bovine colostrum, including the minor protein components, followed by a bioinformatic functional analysis. The proteomic approach based on ProteoMiner technology combined to a highly selective affinity chromatography approach for the immunoglobulins depletion, identified 1786 proteins (medium confidence; 634 when setting high confidence), which were then clustered on the basis of their biological function. Protein networks were then created on the basis of the biological functions or health claims as input. A set of 93 proteins involved in the wound healing process was identified. Such an approach also permits the exploration of novel biological functions of BC by searching in the database the presence of proteins characterized by innovative functions. In conclusion an advanced approach based on an in depth proteomic analysis is reported which permits an explanation of the wound healing effect of bovine colostrum at molecular level and allows the search of novel potential beneficial effects.
Assuntos
Cromatografia de Afinidade/métodos , Colostro/metabolismo , Nanotecnologia/métodos , Proteoma/metabolismo , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Bovinos , Cromatografia Líquida de Alta Pressão/métodos , Colostro/química , Feminino , Leite/química , Leite/metabolismo , Gravidez , Proteoma/químicaRESUMO
The nutrient foramina of the femur and fibula are located in the middle third of the diaphysis of these bones, on the other hand in the tibia it's in the proximal third of its diaphysis. With the objective of complementing the information delivered by other authors, we investigated the location, the number of diaphysis foramines, the distance or position of the foramines in relation to the length of the bone and the proximal epiphysis and the diameter of the bone at the height of the foramen. For that purpose, we used 140 dry bones (50 femora, 50 tibiae and 40 fibulae) of adult subjects of both sexes from the Anatomy Museum Faculty of Medicine, Universidad de La Frontera, Temuco, Chile. The nutrient foramen of the femur was located in the linea áspera in the 72,5 percent (36,25 percent in the interstice, 8,75 in the lateral lip of the linea áspera and 27,5 in the medial lip of the linea áspera), and in the 21,25 percent in the medial surface and in the 6,25 percent in the lateral surface of the diaphysis of the bone. The nutrient foramen of the tibia was found under the soleal line in a 94,33 percent at an average distance of 117.8mm from the intercondylar eminence to the nutrient foramen (in the superior third of the bone), in the soleal line of the bone in 3,77 percent and in the lateral border in 1,88 percent. In the fibula the nutrient foramen was found in the 68 percent in the posterior surface; 14 percent in the medial surface; 12 percent in the interosseous border; 4 percent in the lateral surface and 2 percent in the posterior border. This data could be useful as reference for surgical procedures of the lower limb.
Los forámenes nutricios del fémur y de la fíbula se localizan en el tercio medio de la diáfisis de estos huesos, mientras que, en la tibia se encuentra en el tercio proximal de su diáfisis. Con el objetivo de complementar la información dada por otros autores, estudiamos, la ubicación, número de forámenes, distancia o posición de los forámenes en relación a la longitud del hueso y a la epífisis proximal y el diámetro del hueso a la altura del foramen. Para ello, utilizamos 140 huesos secos (50 fémur, 50 tibias y 40 fíbulas), de individuos adultos, de ambos sexos, procedentes del Museo de Anatomía, Facultad de Medicina, Universidad de La Frontera, Temuco, Chile. El foramen nutricio del fémur se localizaba en la línea áspera en el 72,5 por ciento (36,25 por ciento en el intersticio, 8,75 en el labio lateral de la línea áspera y 27,5 en el labio labio medial de la línea áspera), en el 21,25 por ciento en la cara medial y en el 6,25 por ciento en la cara lateral de la diáfisis del hueso. El foramen nutricio de la tibia fue encontrado bajo la línea del músculo soleo en un 94,33 por ciento, a una distancia promedio de 117,8 mm desde la eminencia intercondílea al foramen nutricio (en el tercio superior del hueso), en la línea del músculo soleo en 3,77 por ciento y en el margen lateral en 1,88 por ciento. En la fíbula el foramen nutricio fue encontrado en el 68 por ciento en la cara posterior; 14 por ciento en la cara medial; 12 por ciento en el margen interóseo; 4 por ciento en la cara lateral y 2 por ciento en el margen posterior. Estos datos sirven como referencia para procedimientos quirúrgicos del miembro inferior.