As pastagens ocupam uma grande extensão territorial no Brasil constituindo a principal fonte de alimento para os ruminantes. Entretanto, seu manejo, ainda é frequentemente realizado de forma extensiva e extrativista, tornando a atividade pouco competitiva. O estudo de estratégias de manejo do pastejo sob lotação contínua contribui para a diversificação e intensificação dos sistemas de produção de ruminantes baseados em pastagens. O objetivo foi descrever e explicar diferenças no potencial produtivo e nas características qualitativas, através de atributos estruturais e químico-bromatológicos da Brachiaria híbrida (Brachiaria ssp.) Convert HD 364 em resposta ao manejo do pastejo sob lotação contínua mimetizada e taxa de lotação variável. O experimento foi conduzido em Piracicaba, no verão de 2013. Foram testadas três intensidades de desfolhação determinadas por alturas de manejo de dossel (10, 25 e 40 cm) em combinação com dois ritmos de crescimento (50 e 250 kg N ha-1 ano-1) num arranjo fatorial (3 x 2) e delineamento em blocos completos casualizados. Variáveis-resposta medidas incluíram massa, acúmulo e taxa de acúmulo de forragem, IAF, composição morfológica e valor nutritivo (Proteína bruta - PB, Fibra insolúvel em detergente neutro- FDN, Digestibilidade in vitro da matéria orgânica - DIVMO) da forragem. Os maiores valores de acúmulo e taxa de acúmulo foram registrados para os dosséis mantidos a 40 cm de altura (12650 kg MS ha-1ano-1 e 110 kg MS ha-1 dia-1, respectivamente) e para dosséis adubados com 250 kg N ha-1(13974 kg MS ha-1ano-1 e 126 kg MS ha-1dia-1, respectivamente). Esse mesmo tratamento apresentou os maiores valores de IAFs (8,1). A massa de forragem não sofreu influência do N, mas respondeu ao aumento da altura e foi maior para os pastos mantidos a 40 cm (14650 kg MS ha-1). Mais N (250 kg ha-1) resultou em maior proporção de folhas (26 %). Maior proporção de colmo (40%) ocorreu no dosséis de 40 cm e adubados com 50 kg N ha-1 e proporções de material morto foram maiores (49%) nos tratamentos de 10 cm de altura e 50 kg de N ha-1. Dosséis de 10 cm adubados com 50 kg N ha-1, embora com menor produtividade, resultaram em maior DIVMO (670 g kg-1) e PB (153 g kg-1 e 165 g kg-1, para 10 cm e 250 kg N ha-1, respectivamente). O teor de FDN foi maior para 40 cm (545g kg-1) e com 50 kg N ha-1(550 g kg-1). A adubação nitrogenada promove ganhos em produtividade e resulta em forragem com melhor valor nutritivo, enquanto que incrementos em altura do dossel promovem maior acúmulo total e taxa média de acúmulo de forragem, até o ponto em que ocorre estabilidade da produção (25 cm), sendo assim o tratamento que apresentou melhor eficiência produtiva aliada a um bom valor nutritivo foi o com 25 cm de altura do dossel e 250 kg ha-1 de N. / Pastures occupy a large area in Brazil and are key to the country\'s livestock industry, which is almost exclusively forage-based. Grazing management, however, is largely empirical in the country, and technical guidelines for managing specific grasses are either scarce or poorly adopted, which hinders the competitiveness of the activity. Establishing sound management guidelines for key grasses can improve animal output and profitability of the livestock enterprise. The objective was to describe and explain differences in the productive potential and the qualitative traits of Convert HD 364 brachiariagrass through the characterization of agronomic and sward structural attributes as well as forage nutritive in response to three grazing intensities and under two growth rates under continuous stocking and variable stocking rate in a mob-grazing experimental protocol. The study was conducted in Piracicaba, in summer of 2013. Experimental paddocks received treatments corresponding to all possible combinations among three sward heights (10, 25, and 45 cm) and two growth rates generated by two N rates (50 and 250 kg N ha-1) split-applied monthly during the experimental period. The experimental design was a randomized complete block with a factorial arrangement. Response variables included forage accumulation and accumulation rate, LAI, plant-part composition of forage on offer, as well as plant-part composition and nutritive value (crude protein - CP, neutral detergent insoluble fiber - NDF digestibility in vitro organic matter - IVDMD) of forage. The highest values of forage accumulation and accumulation rate were recorded for swards kept at 40 cm (12650 kg DM ha-1 yr-1 and 110 kg ha-1 d-1, respectively) and for swards fertilized with 250 kg N ha-1 (13974 kg DM ha-1 yr-1 and 126 kg DM ha-1 day-1, respectively). This same combination resulted the highest LAI (8.1). Forage mass was affected by sward height but not by N rates being higher for 40-cm (14650 kg DM ha-1). Treatments with higher N (250 kg ha-1) resulted in forage with a higher proportion of leaf (26%). Higher percentages of stem (40%) were measured in swards grazed at 40 cm and fertilized with 50 kg N ha-1 while the proportion of dead material was higher (49%) in 10-cm swards receiving 50 kg N ha-1 of N. Swards kept at 10 cm and receiving 50 kg N ha-1, although less productive, resulted in forage with higher IVOMD (670 g kg-1). Forage in 10-cm swards had 153 g kg-1 crude protein and that from swards receiving 250 kg N ha-1 had 165 g kg-1. Neutral detergent fiber concentration was higher in forage from 40-cm swards (545 g kg-1) and from those receiving 50 kg N ha-1 (550 g kg-1), which is also associated with the maturity of the tissues at the time of harvest. Nitrogen fertilization promotes gains in productivity and results in forage with better nutritive value, whereas increases in canopy height promote higher total accumulation and rate of herbage accumulation, up to 25 cm. The treatment that combines high productivity with high nutritive value is 25 cm canopy height and 250 kg ha-1.

No description available.

Animal models have been recognized for the valuable roles they serve in both animal and human medicine. Dogs share many of the same naturally occurring tumors as humans including osteosarcoma, lymphoma, and mammary tumors. In addition, dogs share the same environment as humans, have a shorter lifespan, and often have a quicker progression of disease, making them an attractive model of human disease. Platelets are small anucleate cell fragments that have essential roles in hemostasis, angiogenesis, and wound healing, and, more recently recognized, roles in development, survival, growth, and metastasis of various cancers. Their roles in angiogenesis has proven to be both directly and indirectly linked to tumor growth due to the angiogenic roles they play in the development of tumor blood supply. Being able to study the interactions and mechanisms between platelets and tumor cells at the protein level, through proteomics, would allow great insight into the effects of platelets on tumor cell behavior as well as potential biomarker identification and therapeutic development. The objective of this research is to integrate the roles of canine platelet proteins into a better understanding of the effects and interactions that platelets have with different tumor cells while utilizing the canine model of neoplasms commonly affecting their human counterparts. The first study in this research describes an efficient technique developed for the purification of canine platelets from clinically relevant volumes of whole blood with high platelet recovery and minimal contamination from other blood cells. The second study describes a non-electrophoretic detergent fractionation technique used for the digestion of canine platelet samples for proteomic analysis as well as description of the proteomic findings for the normal canine platelet. Lastly, the third study describes the proteomic analysis of proteins differentially expressed by canine osteosarcoma and mammary tumor cells following incubation with canine platelet lysate in vitro. Overall, findings of this research support the canine model of human cancers and provide comprehensive information regarding canine platelet proteomics as well as novel efficient techniques that aid the future of canine platelet-tumor cell interaction research

This study investigated the effects of the detergent phosphorus ban implemented on January 1, 1988 in Virginia, on the treatment streams of the York River Wastewater Treatment Plant, a biological nutrient removal (BNR) system. Evaluation of the available historical data indicated that the influent phosphorus load entering the plant in the post-ban period had decreased 27% compared to the pre-ban period. The influent phosphorus concentration had decreased 29% from the pre- to the post-ban period. No definitive conclusion could be reached concerning the effects of the influent phosphorus decrease on the treatment efficiency due to operational changes which occurred at the time of the ban implementation. The combination of operational changes and the phosphorus ban resulted in 54% and 59% decreases in the effluent phosphorus load and concentration, respectively. Measurements and modelling of the anaerobic digester contents indicated that several minor changes had occurred in the digester which may be attributable to the phosphorus ban, but the equilibria of the digester regarding phosphorus compounds had not been dramatically altered. This was primarily due to improved operation of the clarifiers and sludge thickeners, which delivered Similar phosphorus loads to the digester in the pre- and post-ban periods. / Master of Science

Increased salinity in freshwater systems – also called the Freshwater Salinization Syndrome (FSS) – can have far-ranging implications for the natural and built environment, agriculture, and public health at large. Such risks are clearly on display in the Occoquan Reservoir – a drinking water source for roughly one million people in the northern Virginia/ National Capital Region. Sodium concentrations in the Occoquan Reservoir are approaching levels that can affect taste and health. The Reservoir is also noteworthy as a flagship example of indirect potable reuse, which further adds complexity to understanding the sources of rising levels of sodium and other types of salinity. To help understand the role residential discharges might play in salinization of the Occoquan Reservoir, a suite of laundry detergent products was identified based upon survey data collected in the northern Virginia region. The ionic compositions of these products were then characterized using ion chromatography and inductively coupled plasma-mass spectrometry to quantify select ionic and elemental analytes. Sodium, chloride, and sulfate were consistently found in appreciable amounts. To comparatively characterize the laundry detergents, principal component analysis was employed to identify clusters of similar products. The physical formulation of the products was identified as a marker for their content, with dry formulations (free-flowing and encapsulated powders) being more enriched in sodium and sulfate. This result was corroborated by comparing nonparametric bootstrap intervals for individual analytes. The study's findings suggest an opportunity wherein consumer choice can play a role in mediating residential salt inputs in receiving bodies such as the Occoquan Reservoir. / Master of Science / Many streams, rivers, and other freshwater systems have become increasingly salty in recent decades. A rise in salinity can be problematic, stressing aquatic life, corroding pipes, and even enhancing the release of more pollutants into the water. This phenomenon, called Freshwater Salinization Syndrome, can threaten such systems' ability to serve as sources of drinking water, as is the case for the Occoquan Reservoir in northern Virginia. Serving roughly one million people, the Reservoir is notable for being one of the first in the country to purposely incorporate highly treated wastewater upstream of a drinking water supply. Despite the Reservoir's prominence, the reasons behind its rising salt levels are not well understood. This study sought to understand the role that individual residences could play when household products travel down the drain and are ultimately discharged into the watershed. Laundry detergents are potentially high-salt products. A survey of northern Virginian's laundry habits was conducted to understand local tastes and preferences. Informed by the survey, a suite of laundry detergents was chemically characterized to measure salt and element concentrations. The detergents were found to have notable amounts of sodium, chloride, and sulfate in particular, with sodium being the most abundant analyte in every detergent. However, not all detergents were equally salty; statistical tools revealed that dry formulations (such as powdered and powder-filled pak detergents) contributed more sodium and sulfate, among other things. This study's findings suggest that laundry detergents could be contributing to Freshwater Salinization Syndrome in the Occoquan Reservoir, and that local consumers' choice of detergents could make a difference.

INTRODUÇÃO: A limpeza prévia de endoscópios rígidos (ER) seguida de desinfecção de nível intermediário com álcool etílico a 70% p/v após o exame de endoscopia nasal é uma prática adotada em muitos serviços de otorrinolaringologia. A literatura atual, no entanto, recomenda a esterilização ou desinfecção de alto nível como o método de descontaminação mais aceito para produtos para saúde classificados como semicríticos. No entanto, há que se fazer distinção entre equipamentos de alta complexidade e sua invasividade como os endoscópios flexíveis com lumens longos e estreitos utilizados na endoscopia digestiva, daqueles de conformação simples sem lumens de baixa invasividade como os endoscópios rígidos utilizados em otorrinolaringologia. OBJETIVO: Avaliar a segurança da desinfecção de nível intermediário com álcool etílico 70% p/v, após limpeza prévia dos endoscópios rígidos utilizados em procedimentos clínicos de endoscopia nasal considerando a carga microbiológica recuperada após o uso. MÉTODO: Imediatamente após a realização do exame, uma gaze úmida foi utilizada para o arraste da carga biológica do endoscópio rígido, gerando as amostras do Controle Positivo e, após a aplicação do POP, um novo arraste para constituir as amostras do Grupo Experimental. Estas gazes foram inicialmente submetidas à sonicação e agitação imersas em soro fisiológico e em seguida a solução foi submetida a uma técnica de extração de carga microbiológica por filtragem por meio de uma Membrana de Celulose de 0,22um de poro que foi, em seguida, semeada nos meios de ágar Sangue, Chocolate, Sabouraud, Löwenstein-Jensen e Tioglicolato. Estes meios ficaram incubados em estufa a 37ºC ± 2ºC e avaliados, no máximo, até por 60 dias conforme o perfil de crescimento dos diferentes microrganismos de interesse; foram analisados de maneira quantitativa e qualitativa para identificação e classificação dos micro-organismos recuperados após as semeaduras. RESULTADO: Os resultados da análise estatística evidenciaram diferença significativa entre Controle Positivo e Grupo Experimental quando comparados em relação à presença de Streptococcus coagulase negativa (p < 0,001), Bacillus spp (p < 0,001) e Staphylococcus aureus (p=0,001). No Controle Positivo, foram encontradas presença desses micro-organismos respectivamente na seguinte frequência: 63,2%, 28,9% e 28,9%, enquanto que, no Grupo Experimental, não foi houve recuperação microbiana alguma. CONCLUSÃO: Os resultados desta pesquisa demonstram a eficiência, na prática diária, da desinfecção de nível intermediário dos endoscópios utilizados na otorrinolaringologia por meio da fricção com álcool etílico 70% p/v por 90 segundos, com protocolo de limpeza prévia / INTRODUCTION: Prior cleaning of rigid endoscopes (REs) followed by intermediate-level disinfection with 70% ethyl alcohol (w/v) after nasal endoscopy is a common practice in many otolaryngology services. Current literature, in turn, recommends high-level sterilization or disinfection as the most accepted decontamination method for health products classified as semi-critical. However, it is necessary to distinguish highly complex equipment according to their invasiveness, e.g., flexible endoscopes with long and narrow lumens used in digestive endoscopy and those with a simple conformation without lumens of low invasiveness, such as rigid endoscopes used in otorhinolaryngology. OBJECTIVE: To evaluate the safety of intermediate-level disinfection with 70% ethyl alcohol (w/v) after cleaning of REs used in clinical procedures of nasal endoscopy considering the microbiological load recovered after use. METHOD: Immediately after the test, a wet gauze was used to drag the biological load from the RE, generating positive control samples; after applying POP, dragging was carried out again to generate samples of the experimental group. These gasses were initially subjected to sonication and shaking while immersed in physiological saline; the solution was then subjected to the microbiological loading technique by filtration through a 0.22-um pore cellulose membrane and then cultivated on blood, chocolate, Sabouraud, Löwenstein-Jensen, and thioglycolate agar media. These media were incubated at 37ºC ± 2ºC and evaluated for up to 60 days, according to the growth profile of the different microorganisms of interest. A quantitative and qualitative analysis was performed for the identification and classification of microorganisms recovered after cultivation. RESULTS: The results of statistical analysis showed a significant difference between the positive control and experimental groups for the presence of coagulase-negative Streptococcus (p < 0.001), Bacillus spp (p < 0.001), and Staphylococcus aureus (p=0.001). In the positive control group, these microorganisms were found in the following proportions: 63.2%, 28.9%, and 28.9%, respectively, whereas in the experimental group, no microorganisms were recovered. CONCLUSION: The results of this study demonstrate the efficiency of the daily practice of intermediate-level disinfection of endoscopes used in otorhinolaryngology by means of treatment with 70% ethyl alcohol (w/v) for 90 seconds, using a previous cleaning protocol

Estudos in situ comumente são conduzidos com pequeno número de animais, existindo poucos trabalhos enfocando o impacto da variabilidade animal sobre seus resultados. O objetivo deste estudo foi explorar o efeito da variabilidade animal sobre as taxas fracionais de degradação (kd) de MS, FDN, FDA e CEL de forragens. Foi conduzido experimento utilizando trinta novilhos Nelore portadores de cânula ruminal (372 ± 16,02 kg P.V.). Dieta 1, todos os animais receberam dieta contendo 100% feno de Coast-cross + sal mineral, ambos ad libitum. Na Dieta 2, todos os animais receberam dieta com 88% de bagaço de cana in natura, 8% de farelo de soja, 2,8% de premix mineral e 1,2% de uréia. Em ambos os períodos foram incubadas amostras de feno de Coast-cross (Fc) (7,31% PB; 78,24% FDN; 40,92% FDA), bagaço de cana-de-açuçar (BAG) (3,04% PB; 90,31% FDN; 69,36% FDA) e feno Tifton-85 (Ft) (13,32% PB; 73,27% FDN; 39,29% FDA) em sacos de poliamida (10 cm x 20 cm; 30 ± 10 &mu;m porosidade) nos tempos 0, 12, 24, 48, 72 e 144 h. Os sacos foram suspensos no saco ventral do rúmen em ordem reversa e removidos simultaneamente no tempo zero. Os teores de MS, FDN, FDA e lignina nos resíduos foram determinados por química líquida e a CEL foi calculada por subtração do teor de lignina da fração do FDA. A cinética de degradação ruminal de MS, FDN, FDA e CEL foram calculadas de acordo com o modelo não linear Y(t) = a + b (1 - e(-kd × t) ); onde t >= 0. As atividades de comportamento ingestivo (ingestão, ruminação, ingestão de água e ócio) foram registradas em intervalos de 10 minutos durante 24 horas. Análise de Componentes Principais foi utilizada para determinar a correlação entre substratos; Análise Fatorial para caracterizar a população de animais (P<0,05); Análise de Agrupamento e Contrastes Multivariados para compor grupos dentro da população (P<0,01) e Análise de Correlação Canônica para determinar correlação entre kd e comportamento ingestivo (P<0,01). Simulação entre número de animais e coeficiente de variação (CV) foi realizada na tentativa de estimar a quantidade adequada de repetições para estes estudos. As médias de kd para MS, FDN, FDA e CEL foram: 3,26; 3,29; 3,44 e 3,54 para Fc; 3,74; 3,88; 4,07 e 4,49 para Ft; e 2,67; 2,68; 2,58 e 2,65 para BAG respectivamente. Foram compostos três níveis de kd para Fc e Ft (alto, médio e baixo) e quatro para BAG (alto, médio alto, médio baixo e baixo). O CV é adequado para mensuração da precisão do ensaio, sendo mais indicado o índice de variação (IV). Em ambas as fases, para todos os substratos, não houve correlação entre kd e comportamento ingestivo. As diferentes correlações de kd entre substratos para as duas fases, demonstra a influência da dieta ofertada sobre o kd. O estudo da variabilidade e o IV indicam que o número de animais é dependente do substrato avaliado, porém o estudo da variabilidade demonstra que os indivíduos que compõe este número são distintos entre dietas. / In situ studies are general carried out with a few number of animals and there are few trials that inquire the impact of animal variability on these results. The aim of this study was to investigate the effect of animal population variability on fractional disappearance rate (kd) of DM, NDF, ADF and cellulose (CEL). Experiment using thirty ruminally canullated Nellore bulls (372 ± 16.02 kg BW) was carried over. In phase 1, all animals received 100% Coast-cross hay with mineral mix ad libitum. In period 2, all animals received a diet containing 88% sugarcane bagasse, 8% soybean meal, 2.8% mineral mix and 1.2% urea (DM basis). In both trials, representative samples of Coast-cross hay (CCH - 7.31% CP, 78.24% NDF, 40.92% ADF), sugarcane bagasse (BAG - 3.04% CP, 90.31% NDF, 69.36% ADF) and Tifton-85 hay (TH - 13.32% CP, 73.27% NDF, 39.29% ADF) were incubated in polyamide-bags (10 cm × 20 cm; 30 ± 10 &mu;m pore size) for 0, 12, 24, 48, 72, and 144 h. Bags were suspended in the rumen ventral sac before at feeding in reverse order and removed simultaneously at time zero. Residual DM, NDF, ADF, and lignin were determined by wet chemistry. Cellulose (CEL) was calculated as ADF minus lignin. Ruminal degradation kinetics of DM, NDF, ADF, and CEL were calculated using a nonlinear model Y(t) = a + b (1 - e(-kd × t) ); where t >= 0. Feeding behavior parameters (eating, ruminating, water drinking and idling) were recorded every 10 minutes during 24 h. Principal component analysis was used to determine correlation of variables among substrates. Factorial Analysis was used to characterize animal population (P<0.05), Cluster with Multivariate Contrasts (P<0.01) to analyze differences in kd groups and Multivariate Canonical correlation analysis (P<0.05) to correlate kd with feeding behavior parameters. A simulation with number of animals was done against coefficient of variation (CV) to determinate optimum number of animals in an in situ assay. The kd (average) of DM, NDF, ADF, and CEL were: 3.26, 3.29, 3.44 and 3.54 for CCH; 3.74, 3.88, 4.07 and 4.49 for TH; and 2.67, 2.68, 2.58 and 2.65 for BAG, respectively. There was no correlation among substrates in both periods. Were found three groups of kd for CCH and TH (high, medium and low) and four groups for BAG (high, medium-high, medium, and low). CV is not adequate to measure experimental precision, instead it, variation index is indicated (VI). In both incubations phases, there was no relationship between feeding behavior and kd, for all incubated forages sources. The kd correlations among substrates between two incubation phases were different, indicating influence of the diet on kd. Variability investigation and VI parameter indicate that the number of animals necessary in these trials is affected by the substrate incubated. However, the variability investigation suggests that probably different animals compose this number.

Estudos in situ comumente são conduzidos com pequeno número de animais, existindo poucos trabalhos enfocando o impacto da variabilidade animal sobre seus resultados. O objetivo deste estudo foi explorar o efeito da variabilidade animal sobre as taxas fracionais de degradação (kd) de MS, FDN, FDA e CEL de forragens. Foi conduzido experimento utilizando trinta novilhos Nelore portadores de cânula ruminal (372 ± 16,02 kg P.V.). Dieta 1, todos os animais receberam dieta contendo 100% feno de Coast-cross + sal mineral, ambos ad libitum. Na Dieta 2, todos os animais receberam dieta com 88% de bagaço de cana in natura, 8% de farelo de soja, 2,8% de premix mineral e 1,2% de uréia. Em ambos os períodos foram incubadas amostras de feno de Coast-cross (Fc) (7,31% PB; 78,24% FDN; 40,92% FDA), bagaço de cana-de-açuçar (BAG) (3,04% PB; 90,31% FDN; 69,36% FDA) e feno Tifton-85 (Ft) (13,32% PB; 73,27% FDN; 39,29% FDA) em sacos de poliamida (10 cm x 20 cm; 30 ± 10 &mu;m porosidade) nos tempos 0, 12, 24, 48, 72 e 144 h. Os sacos foram suspensos no saco ventral do rúmen em ordem reversa e removidos simultaneamente no tempo zero. Os teores de MS, FDN, FDA e lignina nos resíduos foram determinados por química líquida e a CEL foi calculada por subtração do teor de lignina da fração do FDA. A cinética de degradação ruminal de MS, FDN, FDA e CEL foram calculadas de acordo com o modelo não linear Y(t) = a + b (1 - e(-kd × t) ); onde t >= 0. As atividades de comportamento ingestivo (ingestão, ruminação, ingestão de água e ócio) foram registradas em intervalos de 10 minutos durante 24 horas. Análise de Componentes Principais foi utilizada para determinar a correlação entre substratos; Análise Fatorial para caracterizar a população de animais (P<0,05); Análise de Agrupamento e Contrastes Multivariados para compor grupos dentro da população (P<0,01) e Análise de Correlação Canônica para determinar correlação entre kd e comportamento ingestivo (P<0,01). Simulação entre número de animais e coeficiente de variação (CV) foi realizada na tentativa de estimar a quantidade adequada de repetições para estes estudos. As médias de kd para MS, FDN, FDA e CEL foram: 3,26; 3,29; 3,44 e 3,54 para Fc; 3,74; 3,88; 4,07 e 4,49 para Ft; e 2,67; 2,68; 2,58 e 2,65 para BAG respectivamente. Foram compostos três níveis de kd para Fc e Ft (alto, médio e baixo) e quatro para BAG (alto, médio alto, médio baixo e baixo). O CV é adequado para mensuração da precisão do ensaio, sendo mais indicado o índice de variação (IV). Em ambas as fases, para todos os substratos, não houve correlação entre kd e comportamento ingestivo. As diferentes correlações de kd entre substratos para as duas fases, demonstra a influência da dieta ofertada sobre o kd. O estudo da variabilidade e o IV indicam que o número de animais é dependente do substrato avaliado, porém o estudo da variabilidade demonstra que os indivíduos que compõe este número são distintos entre dietas. / In situ studies are general carried out with a few number of animals and there are few trials that inquire the impact of animal variability on these results. The aim of this study was to investigate the effect of animal population variability on fractional disappearance rate (kd) of DM, NDF, ADF and cellulose (CEL). Experiment using thirty ruminally canullated Nellore bulls (372 ± 16.02 kg BW) was carried over. In phase 1, all animals received 100% Coast-cross hay with mineral mix ad libitum. In period 2, all animals received a diet containing 88% sugarcane bagasse, 8% soybean meal, 2.8% mineral mix and 1.2% urea (DM basis). In both trials, representative samples of Coast-cross hay (CCH - 7.31% CP, 78.24% NDF, 40.92% ADF), sugarcane bagasse (BAG - 3.04% CP, 90.31% NDF, 69.36% ADF) and Tifton-85 hay (TH - 13.32% CP, 73.27% NDF, 39.29% ADF) were incubated in polyamide-bags (10 cm × 20 cm; 30 ± 10 &mu;m pore size) for 0, 12, 24, 48, 72, and 144 h. Bags were suspended in the rumen ventral sac before at feeding in reverse order and removed simultaneously at time zero. Residual DM, NDF, ADF, and lignin were determined by wet chemistry. Cellulose (CEL) was calculated as ADF minus lignin. Ruminal degradation kinetics of DM, NDF, ADF, and CEL were calculated using a nonlinear model Y(t) = a + b (1 - e(-kd × t) ); where t >= 0. Feeding behavior parameters (eating, ruminating, water drinking and idling) were recorded every 10 minutes during 24 h. Principal component analysis was used to determine correlation of variables among substrates. Factorial Analysis was used to characterize animal population (P<0.05), Cluster with Multivariate Contrasts (P<0.01) to analyze differences in kd groups and Multivariate Canonical correlation analysis (P<0.05) to correlate kd with feeding behavior parameters. A simulation with number of animals was done against coefficient of variation (CV) to determinate optimum number of animals in an in situ assay. The kd (average) of DM, NDF, ADF, and CEL were: 3.26, 3.29, 3.44 and 3.54 for CCH; 3.74, 3.88, 4.07 and 4.49 for TH; and 2.67, 2.68, 2.58 and 2.65 for BAG, respectively. There was no correlation among substrates in both periods. Were found three groups of kd for CCH and TH (high, medium and low) and four groups for BAG (high, medium-high, medium, and low). CV is not adequate to measure experimental precision, instead it, variation index is indicated (VI). In both incubations phases, there was no relationship between feeding behavior and kd, for all incubated forages sources. The kd correlations among substrates between two incubation phases were different, indicating influence of the diet on kd. Variability investigation and VI parameter indicate that the number of animals necessary in these trials is affected by the substrate incubated. However, the variability investigation suggests that probably different animals compose this number.

Garantir la qualité des produits sanguins est crucial. Les lysats plaquettaires (LP) riches en facteurs de croissance (FC) s’imposent comme le complément idéal pour l’expansion ex vivo des cellules souches, et comme produit thérapeutique pour la régénération cellulaire. L’intérêt est croissant pour les microparticules (MPs) extracellulaires, mais l’expression de phosphatidylsérine à leur surface peut induire des effets thrombotiques et inflammatoires. L’autre risque transfusionnel, la transmission de virus, dont le virus de l’hépatite C (VHC), est maîtrisable par traitements de réduction virale par solvant/détergent (S/D), chauffage, ou nanofiltration. Nous avons étudié des technologies de sécurisation des produits sanguins: (a) élimination des MPs par nanofiltration sur filtres de 75 nm et (b) traitements S/D, chauffage à 56°C ou nanofiltration pour inactiver ou éliminer le VHC. Les informations ont été utilisées pour développer des LP utiles en médecine régénérative. L’un d’eux destiné à la neurorégénération, a été préparé en émettant l’hypothèse qu’un lysat de culot plaquettaire (LCP) enrichi en facteurs neurotrophiques et dépourvu de protéines plasmatiques se montrerait efficace contre les maladies neurodégénératives. Nos résultats montrent que la nanofiltration sur des filtres de 75 nm préserve la composition en protéines plasmatiques, et le pouvoir hémostatique. La nanofiltration retire les MPs et évite, in vitro, la génération de thrombine. Par ailleurs le traitement S/D à 31°C pour 30 minutes élimine le pouvoir infectieux du VHC. Pris globalement les traitements de nanofiltration et S/D apparaissent donc comme des méthodes de choix pour l’amélioration de la sécurité du plasma vis à vis de risques thrombogènes et infectieux. Nous avons ensuite préparé un LCP appauvri en protéines plasmatiques (dont le fibrinogène) et enrichi en un mélange pléiotrope physiologique de FC destiné à l’administration cérébrale. Les analyses par ELISA et par protéomique ont montré qu’un chauffage de 56°C pour 30 min réduisait le contenu en protéines et modifiait favorablement la composition relative en facteurs neurotrophiques. Par ailleurs le chauffage améliore l’action neuroprotectrice et, associé aux traitements S/D et de nanofiltration, contribue à l’inactivation du VHC. Ce LCP exerce une neuroprotection élevée dans des modèles de la maladie de Parkinson (MP) tout à la fois (a) in vitro (cellules LUHMES différentiées en neurones dopaminergiques et exposées au MPP+) et (b) in vivo (souris intoxiquées par MPTP). L’expression de la tyrosine hydroxylase (TH) dans la Substantia nigra pars compacta montre que l’administration intracérébroventriculaire (ICV) ou intranasale (i.n.) apparait comme une option thérapeutique possible des maladies neuro-dégénératives. Les études cellulaires In vitro sur LUHMES et NSC34 ont montré que l’inhibition spécifique des voies signalétiques relayées par AkT et ERK altère l’activité neuroprotectrice du LC. Des événements neuro-inflammatoires pouvant aggraver l’évolution des maladies neurodégénératives, nous avons vérifié que le LCP n’induit pas de marqueurs inflammatoires (COX-2, iNOS) chez des cellules microgliales BV2, et pouvait même diminuer celle de COX-2 après exposition à des lipopolysaccharides. De plus, nous avons identifié que le LCP contenait 1.7 x 1012 MP/mL d’une taille moyenne de 160 nm. Isolées, ces MPs pourraient exercer un rôle neuroprotecteur des cellules LUHMES exposées à des agents neurotoxiques. En conclusion, nos résultats montrent la faisabilité technique à préparer des lysats plaquettaires viro-inactivés pour des usages dans le domaine de la médecine régénérative, y compris comme agent neuroprotecteur du système nerveux central. / Ensuring quality and safety of blood products is crucial. Platelet lysates (PL) rich in growth factors (GFs) have emerged as ideal clinical-grade supplement for ex vivo expansion of mesenchymal stromal cells, and as therapeutic product promote cellular regeneration. Interest for platelet extracellular microparticles (MPs) is growing but expression of phosphatidylserine on their surface may cause thrombotic and inflammatory side effects. Another transfusional risk, transmission of viruses, including hepatitis C virus (HCV), can be fully controlled by dedicated viral reduction methods as solvent/detergent (S/D) or heat treatments, or nanofiltration. We have evaluated technologies to secure therapeutic blood products: (a) removal of MPs by 75nm-nanofiltration and (b) inactivation/removal of HCV by S/D or 56°C heat treatments, or nanofiltration. Data have been used to develop LP of interest for regenerative medicine. In particular, one, targeting neuroregenerative applications, has been prepared based on the hypothesis that a platelet pellet lysate (PPL) enriched in multiple neurotrophic growth factors and depleted of plasma proteins could exert potent neuroprotective actions in neurodegenerative disease models. Our data show that 75 nm-plasma nanofiltration preserved plasma protein biochemical profile, and hemostatic power. Nanofiltration removes MPs and avoids in vitro the generation of thrombin. In addition, the S/D treatment at 31°C for 30 minutes fully inactivates HCV infectivity. Therefore, altogether, nanofiltration and S/D emerge as choice procedures to improve the safety of plasma for thrombogenic and infectious risks. We have then prepared a PPL depleted of plasma proteins (in particular fibrinogen), and rich in a physiological pleiotropic mixture of neurotrophins for brain administration. ELISA and proteomics studies revealed that the heat-treatment at 56°C for 30 min decreased the protein content and favorably modified the relative composition in neurotrophic factors. Heat-treatment improved the neuroprotective activity and, together with S/D and nanofiltration contributed to HCV inactivation. This PPL exerted strong neuroprotective effects in Parkinson’s disease (PD) models (a) in vitro, using LUHMES cells exposed to MPP+ neurotoxin, and (b) in vivo, in mice intoxicated by MPTP neurotoxin. Expression of tyrosine hydroxylase (TH) in the Substantia nigra pars compacta indicated that brain delivery by intracerebroventricular (ICV) or intranasal (i.n.) administration may be a therapeutic option for disease-modifying strategies of neurodegenerative diseases. In vitro studies in LUHMES and NSC34 cells showed that specific inhibition of signal transduction pathways through AkT and ERK influenced PPL neuroprotective function. Since neuro-inflammation detrimentally affects neurodegenerative disorders, we verified that the PPL did not stimulate the release of inflammatory markers (e.g. COX-2, iNOS) by BV2 microglial cells in culture, and could even restrict COX-2 expression when cells were exposed to LPS. In addition, the PPL was found to contain 1.7 x 1012 MP/mL with a mean size of 160 nm. These MPs may exert neuroprotective activity on LUHMES cells exposed to neurotoxins. Altogether, our data demonstrate the technical feasibility of developing virally-safe customized platelet lysate preparations with specific applications for cell therapy and regenerative medicine, in particular as neuroprotective agents of the central nervous system.

When plants are exposed to multiple environmental stress factors, one form of stress can affect the response to another stress. This study used seedlings of a new cultivar of wheat(Triticum aestivum L. cv. 1862), grown under factorial combinations of two levels of ultraviolet-B (UV-B)radiation, two salinity regimes and two levels of iron treatment in chelator-buffered nutrient solutions in a growth chamber. A number of morphological and physiological measurements were made. The accumulation of chlorophyll, UVabsorbing compounds and proline in shoots, as well as phytosiderophores (PSs) in root exudates were measured. Feed value measurements included crude protein, water-soluble carbohydrates, acid detergent fibre and Fe in shoots and roots. After 21 days of stress exposure, results showed that Fe deficiency and NaCl stress generally decreased plant growth and function as well as nutritive value, but increased plant biochemical protection traits such as proline accumulation (16.3 fold under salinity stress) and release of PSs (2.4 fold under Fe deficiency). Interestingly, UV-B radiation affected belowground parameters, inducing a 47% reduction in PS release, together with decreasing root DM by 9% and Fe concentration in roots by 7%. When Fe deficiency and NaCl stress were combined, the results showed a decrease in PS release by 3.5 fold compared to unstressed plants. UV-B radiation synergistically increased UV-absorbing compound levels in combination with Fe deficiency, compared to plants grown under optimal Fe levels. This stress combination also resulted in a cumulative effect by decreasing Fe concentration in shoots and roots. However, salt stress did not interact with UV-B radiation for any of the traits measured. In addition, some three-way interactions were noted, with the Fe x NaCl x UV-B stress combination slightly decreasing PS release and resulting in a cumulative effect by decreasing Fe concentration in roots. In conclusion, this study found that aboveground stress factors such as UV-B can affect important aspects of belowground plant function, and that Fe deficiency can interact with UV-B and salinity stress in modifying plant responses to either stress alone.