Resumo: O presente estudo foi realizado com o objetivo avaliar as características morfogênicas e o acúmulo de forragem do dossel de Brachiaria brizantha cv. Marandu em função de diferentes ofertas de forragem, em regime de desfolha intermitente, ao longo da estação chuvosa. O experimento foi realizado no setor de forragicultura da FCAV, no período chuvoso de 2005/2006. O delineamento experimental foi em blocos ao acaso, utilizando o esquema de parcelas subdivididas, com três repetições. Nas parcelas foi avaliado o efeito das ofertas de forragem (4, 7, 10 e 13 % de OF) e na subparcela o efeito dos ciclos de pastejo (CP). Foram avaliadas as seguintes variáveis: altura do dossel comprimido, massa de forragem, dinâmica de acúmulo de forragem e características morfogênicas e estruturais. Observou-se aumento da altura do dossel comprimido com acréscimo da OF. Da mesma forma foi verificado acréscimo na massa de forragem. Constatou-se aumento da massa de forragem, pós pastejo, com a sucessão dos CP, exceto para OF de 4 %, que se manteve constante (3181 kg/ha) ao longo dos CP. Apenas a taxa de alongamento de colmo, das características morfogênicas sofreu alteração em função da OF, sendo que, esta variável aumentou linearmente com a OF. A densidade de perfilhos vegetativos aumentou linearmente com a intensidade de pastejo, no entanto, a densidade de perfilhos reprodutivos e totais ajustou-se ao modelo quadrático. Observou-se tendência de maior acúmulo de forragem nas OF intermediárias (7 e 10 %). Sendo assim, concluise que OF ao redor de 8 % parece ser a mais indicada para o manejo de pastagem de capim - Marandu em pastejo rotacionado. / Abstract: The present study aimed to evaluate the morphogenetic characteristics and the herbage accumulation of Brachiaria brizantha cv. Marandu swards in function different herbage allowance, managed on intermittent stocking during the summer season. The experiment was carried out at Departamento de Zootecnia, FCAV/UNESP, in the summer period of 2005/2006 year. The experimental design was the complete randomized block, with split plot scheme, and three replications. In the plot the effect of the herbage allowance was evaluated (4, 7, 10 and 13% of HA) and in the split plot the effect of the grazing cycles (GC). There were evaluated the following variables: height of the compressed sward, forage mass, forage accumulation dynamics and morphogenetic and structural characteristics. An Increase of the compressed sward height was observed with increment of HA. Thus, the increase in the herbage mass was verified. An increase herbage mass after grazing was verified with the GC succession, except for 4% of HA, which kept constant (3181 kg/ha) along GC. The stem elongation rate, was the only morphogenetic characteristics altered in function of HA and, this variable increased lineally with HA. The vegetative tiller density increased lineally with the grazing intensity. However, reproductive and total density of tillers was adjusted to the quadratic model. There was a tendency of larger forage accumulation in intermediate HA (7 and 10%). As conclusion the HA around 8% showed the most suitable for grassland management with Marandu-grassin rotational stocking. / Orientador: Ana Claudia Ruggieri / Coorientador: Ricardo Andrade Reis / Banca: Carlos Guilherme Silveira Pedreira / Banca: Patrícia Menezes Santos / Mestre

Orientador: Ana Cláudia Ruggieri / Banca: Carlos Augusto de Miranda Gomide / Banca: Ricardo Andrade Reis / Resumo: O uso de pastagens é a alternativa mais econômica para a alimentação de ruminantes. É de suma importância o conhecimento das características morfogênicas e estruturais dos pastos, uma vez que as mesmas determinam o índice de área foliar do relvado, refletindo diretamente no pastejo pelos animais. O experimento foi conduzido no período de janeiro/2010 a junho/2010, com o objetivo de avaliar as características morfogênicas, estruturais e a dinâmica do perfilhamento do capim tanzânia em diferentes índices de área foliar residual (IAFr), sob pastejo rotacionado por cabras Anglonubiano. Os tratamentos consistiram de três IAF residual (0,8; 1,6 e 2,4). O delineamento experimental utilizado foi o de blocos completos casualizados, em arranjo de parcelas subdividas no tempo, com seis repetições e um total de 18 unidades experimentais. Para determinação do período de descanso foi adotado o critério de 95% de interceptação luminosa (condição de pré-pastejo). Foram avaliadas, no pré e pós pastejo, as alturas do dossel, IL, IAF, massa de forragem e as características morfogênicas e estruturais dos pastos. A altura média, IL e IAF dos dosséis no pré pastejo decresceram com a diminuição do IAFr. No pós pastejo, os IAFr 0,8 e 2,4 apresentaram maior e menor valor de IL, de 35,93 e 62,50%, respectivamente. No pré-pastejo os valores dos componentes de massa dos pastos mantidos sob IAFr 2,4 foram superiores (P<0,10), quando comparados aos pastos sob IAFr 0,8. Houve um aumento nos valores de massa verde seca (8696,85 kg/MS/ha) e massa seca total (12082,00 kg/MS/ha), conforme o ciclo de pastejo, devido principalmente à contribuição da massa seca de colmo (MSC). O período que compreendeu o último ciclo de pastejo teve a TApF 32% menor quando comparada aos primeiro e segundo ciclos de pastejo. O último ciclo de pastejo (Abr/Mai) apresentou TAlF inferior comparadas aos ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The use of pastures is the most economical way to feed ruminants. It is of paramount importance the knowledge of the morphogenetic and structural characteristics of the pasture, since they determine the leaf area index of the sward, is directly reflected by the grazing animals. The experiment was carried from janeiro2010 to June/2010, in order to evaluate the morphogenesis, structure and dynamics of tillering of Guinea grass in different residual LAI under rotational grazing by goats Anglo Nubian. The treatments consisted of three residual rLAI (0.8, 1.6 and 2.4). The experimental design was a complete randomized block, with a split plot arrangement, with six replication and a total of 18 plots. For the determination of rest period has been adopted the criterion of 95% light interception (under pre-grazing). There were evaluated before and after grazing, the sward heights, IL, LAI, herbage mass and the morphogenetic and structural characteristics of the pastures. The average height, canopy LAI and IL pre grazing decreased with decreasing rLAI. In the post grazing, the rLAI 0.8 and 2.4 showed higher and lower LI of 35.93 and 62.50% respectively. In the pre-grazing values of the components of the mass of swards under 2.4 rLAI were higher (P<0,10) when compared to pastures under rLAI 0.8. There was an increase in the values of green dry mass (8696,85 kg / DM / ha) and total dry mass (12082,00 kg / DM / ha) as the grazing cycle, mainly due to the contribution of the dry mass of stem (DMS). The period that included the last period of grazing had LAI 32% lower when compared to the first and second grazing cycles. The last grazing cycle (Apr / May) had markedly lower compared to previous cycles (Mar / Apr). The lifespan of leaves (VPD) was affected by treatments (rLAI) and the grazing season (P<0.10). The rLAI 2.4 resulted in longer life compared to rLAI 0.8 and 1.6. No significant rLAI ... (Complete abstract click electronic access below) / Mestre

O objetivo do atual trabalho, foi correlacionar os eventos celulares e teciduais com a expressão das proteínas VEGF, BMP-7, RANKL e OPG durante a osteogênese ectópica e ortotópica, induzida pela matriz óssea (MO) e dentinária (MD) alogênica. Matrizes alogênicas desmineralizada em HCl a 0,6N, obtidas de fêmur e incisivo de ratos, fori implantada entre as fáscias musculares da coxa e em defeito trans-ósseo de 8mm de diâmetro nos ossos parietais. As análises radiográfica e histomorfométrica da neoformação óssea e, a imunohistoquímica e o western blotting para as proteínas VEGF, BMP, RANKL e OPG, mostraram que: a) o volume da região do enxerto nos sítios ortotópicos reduziu 19% em 42 dias; b) em ambos tipos de enxerto e locais de implantação, ocorreu formação de tecido cartilaginoso e ósseo; c) nos sítios intramusculares, a reabsorção da matriz alogênica e a remodelação do tecido cartilaginoso, ósseo e medular foi mais acelerado, em relação a implantação ortotópico; d) o aumento na densidade de volume dos vasos sanguíneos e no número de osteoblastos/osteócitos e osteoclastos ocorreu simultaneamente e estava associado à maior reabsorção da matriz alogênica e à formação do tecido medular (hematopoiético); e) as proteínas VEGF, BMP-7, RANKL, OPG foram expressas em condrócitos, osteoblastos ativos, osteócitos recém aprisionados na matriz e em células estromais próximas aos osteoblastos ou às áreas da matriz alogênica reabsorvida; e f) a expressão das proteínas VEGF, BMP-7, RANKL e OPG foi maior no grupo MO. O pico de expressão dessas proteínas ocorreu nos períodos de 14 aos 21 dias no grupo da MO e 21 e 28 dias no grupo da MD. Concluímos que, a capacidade osteoindutora da matriz alogênica desmineralizada está relacionado a origem da matriz e ao sítio de implantação e que, as proteínas VEGF, BMP-7, RANKL e OPG estão associadas a maior reabsorção da matriz implantada, promovendo uma rápida e contínua liberação dos morfógenos contidos em seu interior que, induzem temporal e espacialmente a formação óssea/medular. / The aim of the present work was to correlate the cellular and tissue events with the expression of VEGF, BMP-7, RANKL and OPG during ectopic and orthotopic osteogenesis, induced by bone and dentin allogeneic matrix. Allogenic matrices obtained from femur and incisor of rats and demineralized in 0.6 N HCl were implanted into a intramuscular pocket and a 8mm-diameter bone defect in the skull. The radiographic and histomorphometric analysis of new bone formation, and immunohistochemistry and western blotting for VEGF, BMP, RANKL and OPG proteins, showed that: a) the total volume of the graft region in orthotopic site decreased 19% at 42 days b) in both graft types and implantation sites occurred formation of cartilaginous and bone tissues, c) in intramuscular sites, the resorption of allogenic matrix and remodeling of the new formed cartilage and bone were faster, in relation to orthotopic implantation sites; d) the increase in the volume density of blood vessels and in the number of osteoblasts/osteocytes and osteoclasts occurred simultaneously and was associated with greater reabsorption of the allogenic matrix and hematopoietic bone marrow formation; e) VEGF, BMP-7, RANKL, OPG proteins were expressed in chondrocytes, active osteoblasts, newly osteocytes confined and stromal cells located near the osteoblasts or in the surface of the reabsorbed matrix; and f) the VEGF, BMP-7, RANKL and OPG expression was higher in MO grafts than in the MD. The peak of expression of these proteins each occurred at 14 and 21 days in MO and 21 and 28 days in MD. We concluded that, the osteoinductive capacity of allogeneic demineralized matrix is related to matrix origin and implantation site and that the VEGF, BMP-7, RANKL and OPG proteins are associated with greater reabsorption of the implanted matrix, promoting rapid and continuous matrix-release morphogens that induces spatially and temporally the bone and bone marrow formation.

Para testar a hipótese de que a desmineralização in situ das superfícies de contato enxerto-leito, e a forma como o enxerto é estabilizado ao leito, podem influenciar os mecanismos envolvidos na consolidação do enxerto, fragmentos ósseos de 10 mm de diâmetro foram removidos das metáfises proximais tibiais de 36 coelhos (Oryctolagus Cuniculus) e transplantados para uma área adjacente. Na tíbia esquerda dos animais, as superfícies de contato do enxerto e do leito hospedeiro foram desmineralizadas com ácido cítrico pH 1,0 por 3 minutos antes dos enxertos serem fixados ao leito. Na tíbia direita, o transplante do bloco ósseo não foi precedido de desmineralização. Metade dos enxertos foi imobilizada sobre o leito pela superposição de uma membrana não reabsorvível de politetrafluoretileno colada com cianoacrilato ao leito à distância da interface enxerto-leito. A outra metade dos enxertos foi fixada por um parafuso de titânio no centro do enxerto. Assim, foram formados 4 grupos de estudo: membrana (M), membrana + ácido (MA), parafuso (P) e parafuso + ácido (PA). Três animais de cada grupo forneceram espécimes para análise microscópica quantitativa e qualitativa aos 15, 30 e 45 dias de pós-operatório. A análise qualitativa demonstrou que não houve formação óssea na interface em nenhum espécime aos 15 dias e que nos demais períodos, em todos os grupos, a quantidade de tecido ósseo neoformado na interface e seu estágio de maturação aumentaram com o tempo. Ambos os métodos de fixação empregados foram eficientes em manter os enxertos em posição, porém a membrana promoveu menor reabsorção da estrutura do enxerto. A análise quantitativa computadorizada revelou que, aos 30 dias, os grupos MA e PA apresentaram maior área de formação óssea na interface (71,34 ± 12,03%; 56,74 ± 2,15% respectivamente) em relação aos grupos M e P (51,75 ± 11,02%; 43,95 ± 4,05% respectivamente) e superfícies de consolidação óssea mais extensas (93,41 ± 5,95%; 93,73 ± 4,96% respectivamente) do que os grupos sem tratamento ácido (73,49 ± 7,7%; 73,77 ± 11,77% respectivamente para M e P), sendo essas diferenças estatisticamente significantes (p<0,05). Aos 45 dias de pós-operatório, os grupos MA e PA (71,18 ± 8,9%; 58,97 ± 3,97% respectivamente) apresentaram resultados superiores aos grupos M e P (59,78 ± 11,28%; 46,08 ± 3,53% respectivamente) em relação à área de neoformação óssea na interface, porém essa diferença não foi significativa. Concluiu-se que a desmineralização ácida das superfícies contactantes nos enxertos ósseos autógenos em bloco na tíbia de coelhos promoveu a osteogênese na interface enxerto-leito e acelerou a consolidação dos enxertos. Além disso, quando o tratamento ácido foi associado ao uso de membrana como método de fixação, a consolidação e grau de reabsorção óssea foram otimizados. / In order to test the hypothesis that the demineralization \"in situ\" of contacting surfaces of bone graft/bone bed and the fixation method used for graft stabilization can influence the mechanisms involved in the consolidation of the graft, bone fragments of 10 mm in diameter were removed from the proximal tibial metaphysis of thirty-six male rabbits (Oryctolagus Cuniculus) and transplanted to an adjacent area. In the left tibia of the animals, the contacting surfaces of the graft and host bed were demineralized with citric acid pH 1.0 for 3 minutes before the grafts were fixed to the receptor bed. In the right tibia, the bone block transplantation was not preceded by demineralization. Half of the grafts were immobilized on the bone bed by a nonresorbable polytetrafluoroethylene membrane glued with cyanoacrylate adhesive to the host bed distant from the bone graft-bone bed interface. The other half of the grafts were fixed by a titanium screw in the center of the graft. Thus, four groups were formed: membrane (M), membrane + acid (MA), screw (P) and screw + acid (PA). Three animals from each group provided specimens for quantitative and qualitative microscopic analysis at 15, 30 and 45 days postoperatively. Qualitative analysis showed no significant bone formation at the interface in any specimen of the groups at 15 days and on the other periods in all groups, the amount of newly formed bone at the interface as well as the stage of bone maturation increased with time. Both fixation methods were effective in maintaining the graft in position, but the membrane resulted in less resorption of the graft. Quantitative analysis, performed by means of a computer program for image analysis, showed that at day 30, groups MA and PA, showed greater area of bone formation at the interface (71.34 ± 12.03%; 56.74 ± 2 15%) than groups M and P (51.75 ± 11.02%, 43.95 ± 4.05%) and more osseointegrated bone surfaces (93.41 ± 5.95%, 93.73 ± 4.96%) than those without acid treatment (73.49 ± 7.7%, 73.77 ± 11.77%), and these differences were statistically significant. At 45 days postoperatively MA and PA groups (71.18 ± 8.9%, 58.97 ± 3.97%) showed better results than the M and P groups (59.78 ± 11.28%, 46 , 08 ± 3.53%) compared to the area of new bone formation at the interface and osseointegrated surfaces, but these differences were not significant. It was concluded that the acid demineralization of contacting surfaces in autogenous onlay bone grafts in the tibia of rabbits promotes osteogenesis in bone graft-host bed interface and accelerates the consolidation of the grafts. Furthermore, the association of this surface treatment to the use of membrane/cyanoacrylate fixation method, optimizes the results regarding consolidation and degree of bone graft resorption.

Kiso H, Takahashi K, Saito K, Togo Y, Tsukamoto H, et al. (2014) Interactions between BMP-7 and USAG-1 (Uterine Sensitization-Associated Gene-1) Regulate Supernumerary Organ Formations. PLoS ONE 9(5): e96938. / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18545号 / 医博第3938号 / 新制||医||1006(附属図書館) / 31445 / 京都大学大学院医学研究科医学専攻 / (主査)教授 斎藤 通紀, 教授 戸口田 淳也, 教授 妻木 範行 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

There are approximately 600,000 cases of delayed or aberrant fracture healing in people each year, with a small subset of these fractures experiencing disunion. Dietary phosphate deficiency has been shown to impair oxidative phosphorylation and decrease BMP-2 mediated chondrogenic differentiation during fracture healing. Prior studies using pre-committed chondro-progenitor ATDC5 cell line grown in phosphate deficient media showed that energy consumption was linked to protein production and collagen hydroxylation but inversely related to matrix mineralization. The goal of this study was to further define the relationship between energy consumption and BMP-2 mediated stem cell chondrogenic differentiation and further examine how dietary phosphate, and promotion of collagen hydroxylation via ascorbate availability effected these processes. C3H10T1/2 murine cells, a multi-potential cell line, were expanded in pre-differentiation growth medium (DMEM with 10% FBS and 1% Pen/Strep). Once cells reached 60% confluence (day 0), they were grown in differentiating media (α-MEM with 5% FBS and 1X insulin-transferrin-selenium) containing either 100% (1mM) or 25% (0.25mM) inorganic phosphate (Pi), ± 200ng/mL BMP-2(BMP), and ±0.2 mM L-ascorbic acid (AA). In total, there were 8 groups with varying combinations of these three substances. Intracellular lipid, total DNA, protein, and hydroxyproline (HP) content were examined. Chondrocyte gene expression (Col2a1, Acan, ColXa1) and adipocyte gene expression (Pparg, Plin1, Ucp1) were measured to check for cell lineage commitment and specific differentiation of the C3H10T1/2. All measurements were acquired at day 8. The +BMP differentiation media groups contained significantly less DNA content and more protein content than the –BMP differentiation media groups (both p<0.0001). There was also a significant interaction between phosphate and ascorbic acid treatment (p=0.0296), with 25% Pi +AA groups producing significantly more protein than 100% Pi +AA groups. Hydroxyproline production was not different in 100% Pi or 25% Pi conditions (p=0.2951). AA presence in culture media led to greater HP production than culture media lacking AA (p=0.0035) There was a trend of an interaction between phosphate content and AA availability (p=0.0744). 100% Pi ±AA groups produced significantly different amounts of HP while 25% Pi ±AA groups did not produce significantly different amount of HP. Col2a1, Acan, and ColXa1 expression were all increased in +BMP groups. Ascorbic acid treatment groups expressed significantly more Col2a1and Acan than –AA groups. 100% Pi media led to greater Acan expression over 25% Pi groups (p=0.0009), whereas 25% Pi media trended to lead to greater ColXa1 expression over 100% Pi groups (p=0.0734). Pparg and Plin1 expression were increased in the 25% Pi condition. There were no significant differences in expression of Ucp1. C3H10T1/2 cells were significantly affected by phosphate concentration, BMP-2 treatment, and ascorbic acid supplementation. Phosphate deficiency hindered maturation of early chondrocytes into proliferating chondrocytes while also promoting MSC differentiation into the adipocyte cell lineage. Hypertrophic chondrocyte expression was decreased in phosphate deficient media, which may coincide with increased protein production observed in low phosphate conditions. BMP-2 promoted chondrogenesis which resulted in increased protein production. Whereas, lack of ascorbic acid in cell culture media led to decreased hydroxyproline production.

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13031号 / 論医博第2113号 / 新制||医||1016(附属図書館) / 32989 / (主査)教授 妻木 範行, 教授 安達 泰治, 教授 開 祐司 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

Over the past 5 years, the use of titanium implants as temporary anchorage devices (TADs) has become an important tool in clinical orthodontic practices. The use of TADs have provided orthodontists a way of moving teeth against fixed objects rather than against the surrounding teeth, which tend to counteract desired motion. At present, viable attachment of TADs involves direct insertion through gingival tissue and piercing of the bone. Surface modifications such as sandblasted and acid-etched treatment or bone morphogenetic protein surface treatment, however, can be applied to the TADs to promote enhanced osseointegration, thereby allowing the TADs to serve as stable anchors while avoiding bone puncture. In this study, a comparison was made between sandblasted/acid-etched TADs and sandblasted/acid-etched/recombinant human bone morphogenetic protein-2 (rhBMP-2) treated TADs to determine whether rhBMP-2 promotes enhanced osseointegration. A total of 10 rats (4 controls and 6 treated with rhBMP-2) were used in the study, with 1 TAD placed on the skull of each rat. At the end of 6 weeks, the animals were euthanized by carbon dioxide asphyxiation, and bone blocks, each containing a TAD, were prepared for histological examination and biomechanical characterization. The results of this study showed that TADs treated with rhBMP-2 had greater bone formation at the bone-implant interface and an increase in total implant stability.

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