Avaliação da incorporação de enxerto osseo homogeno fresco congelado em humanos: análise histologica e imuno-histoquímica

Allogeneic, fresh-frozen bone has been used in order to replace bone autografts. However, its osteoinduction and osteoconduction properties are not well-defined in the scientific literature. This work aimed to evaluate samples of homogenous bone grafts in humans by qualitative histological and immunohistochemical analysis. For this, ten pre-selected patients underwent surgical augmentation of bone defects. The homogenous fresh frozen block bone graft was stabilized and fixed by bicortical screws. After six months, the reopening procedure was performed for installation of osseointegrated implants. At this time surgical bone graft samples were removed by means of drill trephine. The samples were fixed in 10% formalin, processed with decalcified paraffin, and stained with hematoxylin and eosin. Immunohistochemistry was performed for the expression of Caspase 3 enzyme. The slides were brought to light microscopy for qualitative histology and immunohistochemistry. The results showed non-vital bone tissue, with few areas of deposition of new bone formation on the amorphous matrix, presence of chronic inflammatory infiltrate with areas of osteomyelitis, and expressive immunolabeling of Caspase 3. Given the methods employed and the results it was concluded that the allograft fresh-frozen block is not incorporated into the recipient bed after a healing period of six months.

Uso de aptâmeros na sexagem de sêmen bovino

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Utilização do choque osmótico na avaliação da viabilidade de sêmen criopreservado de ovinos

This study evaluated the use of hipoosmotic swelling test (HOST) with deionized water (0 mOsmol), as a method of post thaw ram semen evaluation and correlate their findings with different techniques of semen evaluation. Therefore, twenty semen samples of 20 different adult rams were assessed as for kinetic sperm parameters through computerized system (IVOS 12, Hamiton Thorn Biosciences, Beverly, MA, EUA) and subjective analysis. The sperm membranes viability was carried out by the association of fluorescent probes (propidium iodide, JC-1 and FITC-PSA). The structural integrity of the plasma membrane was also studied through supravital test with eosin and the functional integrity of membrane evaluated by doing the hipoosmotic swelling test with deionized water (0 mOsmol), in the following proportions: One part of semen for 10 (HOST 10), 50 (HOST 50) and 100 (HOST 100) parts of water. After semen dilution in the different proportions it was fixed in formalin-buffered saline and analyzed with regard to percentage of HOST reactive sperm (bent/coiled). The percentage of reaction obtained for HOST 10 (33,1%); HOST 50 (32,8%) and HOST 100 (31,8%) did not differ significantly. HOST 10 presented positive correlation with the plasma membrane integrity by the EOS (r = 0,80; p < 0,05). Positive correlations between HOST 50 and HOST 100 with sperm subpopulation with membrane integrity by fluorescence were observed (r = 0,83 and r = 0,85; p < 0,01). The findings suggest that the HOST with deionized water can provide additional information for post thawing ram sperm viability evaluation.

Utilização de sistemas de refrigeração de sêmen equino na estabilização das amostras seminais previamente à congelação

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Termografia digital por infravermelho do escroto e qualidade do sêmen em touros nelore (bos taurus indicus)

Aimed to study the relationship between scrotal temperature on semen quality by use of infrared thermography in Nelore bulls bred extensively. There were scrotal thermography and semen collection. Climatic data were collected by means of globe thermometer. The thermograms of the scrotum were obtained by digital infrared thermography. We carried out ejaculates through electroejaculation for analysis of quantitative and qualitative characteristics. For the data, we used analysis of variance and subsequently applied the Tukey test at 5%. (p <0.05) between animals for sperm vigor, sperm motility between animals and between crops, sperm concentration, total sperm and total viable sperm between groups, between harvests for the scrotum temperatures and climate data. It was concluded that the surface temperatures of the scrotum and climatic factors influence the quality of semen. Thermography is recommended as a complementary test in the evaluation of reproductive bulls.

Correlação da circunferência escrotal, citologia testicular e parâmetros do sêmen do epidídimo de touros

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

Estudo sobre a redução do estresse oxidativo em sêmen equino a partir da adição de quercetina nos diluentes de refrigeração e congelação

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

Efeito da centrifugação e filtragem do sêmen bovino sobre a criopreservação espermática

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo sobre a redução do estresse oxidativo em sêmen equino a partir da adição de quercetina nos diluentes de refrigeração e congelação

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

Efeito da centrifugação e filtragem do sêmen bovino sobre a criopreservação espermática

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

O reflexo da nutrição na qualidade do sêmen em reprodutores suínos-suplementação com vitamina A e selênio orgânico

Os organizadores autorizam a reprodução total ou parcial deste trabalho, para qualquer meio convencional ou eletrônico, para fins de estudo e pesquisa, desde que citada a fonte.

Congelación y conservación del semen en la especie caprina mediante la utilización de ultracongeladores de -152ºC: Tasa de fertilidad tras inseminación con semen congelado por diferentes protocolos de crioconservación

[ES] El análisis de los distintos parámetros evaluados para determinar la calidad seminal de siete machos cabríos de raza Majorera, demuestra que la congelación del semen mediante ultracongeladores de -152 ºc es igual de efectiva que la congelación mediante nitrógeno líquido, técnica que se ha venido usando desde hace ya muchos años. se observa cómo la calidad seminal in vitro tras la descongelación no varía en los diferentes períodos de congelación testados (1, 30 y 180 días), así mismo, el porcentaje de gestaciones obtenido a partir de la inseminación artificial en 40 hembras, es similar en ambos protocolos de congelación para muestras congeladas durante un período de un mes

Efecto de la adición de ingredientes funcionales en el comportamiento reológico y la textura de puré de patata (Cv. Kennebec) fresco y congelado

El interés creciente en encontrar alimentos precocinados congelados que se asemejen a productos naturales, capaces de superar un procesado con el menor daño, ha generado un aumento en el estudio de nuevos productos en este campo de la investigación. Las características de cada matriz alimentaria, la composición y estructura de los ingredientes, así como el efecto de las interacciones entre ellos, modifica la textura, estructura y las propiedades físicas y sensoriales del alimento, así como su aceptación por el consumidor. En este contexto, la investigación realizada en esta tesis doctoral se ha llevado a cabo en puré de patata considerado como una matriz alimentaria semisólida y se ha centrado en analizar los efectos de la concentración y modificación de la composición en las propiedades reológicas y de textura, en las propiedades físico-químicas y estructurales, así como en los atributos sensoriales de los purés de patata cuando a estos se le añaden diferentes ingredientes funcionales como fibra de guisante, inulina, aceite de oliva, aislado de proteína de soja, ácidos grasos omega 3 y/o sus mezclas. Para ello, se han realizado cuatro estudios donde se determinan las propiedades reológicas mediante ensayos dinámicos oscilatorios y en estado estacionario, los parámetros instrumentales de textura mediante ensayos de extrusión inversa y de penetración cónica, además de los cambios estructurales a través de cromatografía iónica con detector de pulsos amperométrico, cromatografía de gases con detector de ionización de llama y microscopía electrónica de barrido. Conjuntamente, se han evaluado los atributos sensoriales de los diferentes purés generando los descriptores que mejor definen la calidad sensorial del producto, utilizando un panel de jueces entrenados y valorándose la aceptación global de los nuevos productos mediante un panel de consumidores. En un primer estudio, el puré de patata natural congelado elaborado con crioprotectores se enriqueció con fibra dietética insoluble (fibra de guisante), fibra dietética soluble (inulina) y sus mezclas. La fibra de guisante influyó significativa y negativamente en la textura del puré de patata, percibiéndose en el producto un incremento de la dureza y de la arenosidad, mientras que la inulina produjo un ablandamiento del sistema. En un segundo estudio, el puré de patata natural fresco y congelado/descongelado elaborado con y sin crioprotectores, se enriqueció con fibra dietética soluble (inulina), aceite de oliva virgen extra y sus mezclas. La adición de estos dos ingredientes generó un ablandamiento de la matriz del sistema, produciéndose, sin embargo, un efecto sinérgico entre ambos ingredientes funcionales. La inulina tuvo un efecto más significativo en la viscosidad aparente del producto, mientras que el aceite de oliva virgen extra afectó más significativamente a la pseudoplasticidad, al índice de consistencia y a la viscosidad plástica del mismo. El proceso de congelación y descongelación utilizado favoreció la reducción del tamaño de las partículas de inulina haciéndolas imperceptibles al paladar, obteniéndose productos más cremosos y con mayor aceptabilidad global que sus homólogos frescos. En un tercer estudio, el puré de patata natural fresco y congelado/descongelado elaborado con crioprotectores se enriqueció con mezclas de fibra dietética soluble (inulina) y aislado de proteína de soja. Los resultados demostraron que el ciclo de congelación y descongelación realizado no afecta el grado de polimerización de la inulina. La estructura química de la inulina tampoco se vio afectada por la incorporación de la soja. El proceso de congelación/descongelación, así como la adición de concentraciones altas de inulina y bajas de aislado de proteína de soja, favorecen la disminución de la contribución de la componente viscosa en las propiedades viscoelásticas del puré de patata. La cremosidad fue el único atributo sensorial que presentó una correlación lineal significativa entre las puntuaciones otorgadas por panelistas entrenados y no entrenados. Por último, se elaboró un puré de patata natural fresco y congelado/descongelado optimizado con crioprotectores y enriquecido con la suma de ácido docosahexaenoico (DHA, C22:6 n-3) y ácido eicosapentaenoico (EPA, C20:5 n-3) y con ácido α-linolénico (ALA, C18:3 n-3) microencapsulados. El ciclo de congelación y descongelación no afectó al perfil de ácidos grasos del puré de patata. La adición de omega 3 procedente de aceites de lino y pescado microencapsulados mejora los indicadores nutricionales que definen la calidad de la grasa, obteniéndose un producto más saludable. ABSTRACT The growing interest in finding frozen precooked products that are like a natural product and capable of withstanding initial processing with minimum damage and remaining stable during preservation and reheating prior to consumption has generated an increase in studies of new products in this field of research. The characteristics of each food matrix, the composition and structure of the ingredients and the effect of interactions between them alter the texture, structure and physical and sensory properties of the food product and its acceptance by the consumer. In this context, the research conducted in this doctoral thesis was carried out on mashed potato, considered as a semi-solid food matrix, and focused on analysing the effects of concentration and modification of the composition of the mashed potato matrix on the rheological and textural properties, physicochemical and structural properties and sensory attributes of mashed potato when various functional ingredients are added to it, such as pea fibre, inulin, olive oil, soy protein isolate, omega 3 fatty acids and/or mixtures of these ingredients. Four studies were conducted for this purpose. Rheological properties were determined by oscillatory dynamic tests and stationary state tests, and instrumental texture parameters by backward extrusion and cone penetration tests. Structural changes were studied by ion chromatography with pulsed amperometric detector, gas chromatography with flame ionisation detector and scanning electron microscopy. The sensory attributes of the various mashed potato mixtures were evaluated by generating the descriptors that best defined the sensory quality of the products and using a panel of trained judges, and overall acceptance of the new products was evaluated by a panel of consumers. In the first study, frozen natural mashed potato incorporating cryoprotectants was enriched with insoluble dietary fibre (pea fibre), soluble dietary fibre (inulin) and mixtures of the two. Pea fibre had a significant negative influence on the texture of the mashed potato, producing an increase in hardness and granularity, whereas inulin produced a softening of the system. In the second study, fresh and frozen/thawed natural mashed potato prepared with and without cryoprotectants was enriched with soluble dietary fibre (inulin), extra virgin olive oil and mixtures of the two. The addition of these two ingredients generated softening of the matrix of the system, but a synergic effect between the two functional ingredients was produced. Inulin had a more significant effect on the apparent viscosity of the product, whereas extra virgin olive oil had a more significant effect on its pseudoplasticity, consistency index and plastic viscosity. The freezing and thawing process that was used contributed to a reduction in the size of the inulin particles, making them imperceptible to the palate and producing creamier products with greater overall acceptability than their fresh equivalents. In the third study, the fresh and frozen/thawed natural mashed potato incorporating cryoprotectants was enriched with mixtures of soluble dietary fibre (inulin) and soy protein isolate. The results showed that the freezing and thawing process that was performed did not affect the degree of polymerisation of the inulin. The chemical structure of the inulin was also not affected by the incorporation of soy. The freezing and thawing process and the addition of high concentrations of inulin and low concentrations of soy protein isolate favoured a decrease in the contribution of the viscous component to the viscoelastic properties of the mashed potato. Creaminess was the only sensory attribute that presented a significant linear correlation between the scores given by trained and untrained panellists. Lastly, fresh and frozen/thawed natural mashed potato optimised with cryoprotectants was prepared and enriched with the sum of docosahexaenoic acid (DHA, C22:6 n-3) and eicosapentaenoic acid (EPA, C20:5 n-3) and with α-linolenic acid (ALA, C18:3 n-3), microencapsulated. The freezing and thawing process did not affect the fatty acid profile of the mashed potato. The addition of omega 3 obtained from microencapsulated linseed and fish oils improved the nutritional indicators that define the quality of the fat, producing a healthier product.