Association between glycemic index and glycemic load and the risk of incident coronary heart disease among whites and African Americans with and without type 2 diabetes: The Atherosclerosis Risk in Communities study

Several studies have examined the association between high glycemic index (GI) and glycemic load (GL) diets and the risk for coronary heart disease (CHD). However, most of these studies were conducted primarily on white populations. The primary aim of this study was to examine whether high GI and GL diets are associated with increased risk for developing CHD in whites and African Americans, non-diabetics and diabetics, and within stratifications of body mass index (BMI) and hypertension (HTN). Baseline and 17-year follow-up data from ARIC (Atherosclerosis Risk in Communities) study was used. The study population (13,051) consisted of 74% whites, 26% African Americans, 89% non-diabetics, 11% diabetics, 43% male, 57% female aged 44 to 66 years at baseline. Data from the ARIC food frequency questionnaire at baseline were analyzed to provide GI and GL indices for each subject. Increases of 25 and 30 units for GI and GL respectively were used to describe relationships on incident CHD risk. Adjusted hazard ratios for propensity score with 95% confidence intervals (CI) were used to assess associations. During 17 years of follow-up (1987 to 2004), 1,683 cases of CHD was recorded. Glycemic index was associated with 2.12 fold (95% CI: 1.05, 4.30) increased incident CHD risk for all African Americans and GL was associated with 1.14 fold (95% CI: 1.04, 1.25) increased CHD risk for all whites. In addition, GL was also an important CHD risk factor for white non-diabetics (HR=1.59; 95% CI: 1.33, 1.90). Furthermore, within stratum of BMI 23.0 to 29.9 in non-diabetics, GI was associated with an increased hazard ratio of 11.99 (95% CI: 2.31, 62.18) for CHD in African Americans, and GL was associated with 1.23 fold (1.08, 1.39) increased CHD risk in whites. Body mass index modified the effect of GI and GL on CHD risk in all whites and white non-diabetics. For HTN, both systolic blood pressure and diastolic blood pressure modified the effect on GI and GL on CHD risk in all whites and African Americans, white and African American non-diabetics, and white diabetics. Further studies should examine other factors that could influence the effects of GI and GL on CHD risk, including dietary factors, physical activity, and diet-gene interactions. ^

The eukaryotic cellular stress response: Biochemical and genetic analyses in Saccharomyces cerevisiae

All cells must have the ability to deal with a variety of environmental stresses. Failure to correctly adapt to and/or protect against adverse stress conditions can lead to cell death. In humans, stress response defects have been linked to a number of neurodegenerative diseases and cancer, underscoring the importance of developing a fundamental understanding of the eukaryotic stress response.^ In an effort to characterize cellular response to high temperature stress, I identified and described one member of a novel gene family— RTR1. I show that the RTR1 gene and its protein product genetically and biochemically interact with core subunits of the RNA polymerase II enzyme. Appropriately, loss of RTR1 results in defective transcription from multiple promoters. These data provide evidence that Rtr1, which is essential under stress conditions, acts as a key regulator of transcription.^ In addition to transcriptional regulation, cells deal with many stressors by inducing molecular chaperones. Molecular chaperones are ubiquitous in all living cells and bind unfolded or damaged proteins and catalyze refolding or degradation. Hsp90 is a unique chaperone because it targets specific clients—typically signaling proteins—for maturation. While it has been shown that Sse1, the yeast Hsp110, is a critical regulator of the Hsp90 chaperone cycle, this work describes the molecular basis for that regulation. I show that Sse1 modulates Hsp90 function through regulation of Hsp70 nucleotide exchange. Further, Hsp110-type nucleotide exchange factors (NEFs) appear to have a specific role in modulating Hsp90 function in this manner. Finally, in addition to Hsp110, the eukaryotic cytosol contains two other types of Hsp70 NEF: Snl1 (BAG-domain protein) and Fes1 (HspBP1-like protein). I investigated the cellular roles of these NEFs to better understand the reason that eukaryotic cells contain three distinct protein families that perform the same biochemical function. I show that while cytsolic Hsp70 NEFs have some degree of functional overlap, they also exhibit striking divergence. Taken together, the work presented in this dissertation provides a more detailed understanding of the eukaryotic stress response. ^

Assessing the Environmental Protection Agency's guidelines for cumulative risk assessment: Are they suitable for state agencies and EPA regional offices?

In the field of health risk analysis, cumulative risk assessment (CRA) is a necessary, although undeniably more complex approach to understanding the mixture of stressors, whether chemical or psychosocial, that exist in our environment, in all the pathways through which the chemicals may evolve—air, soil, or water, as well as the accumulation of these exposures over time. Related, or attached to the developing awareness of scientists understanding this mix of combined health effects is the burgeoning of the environmental justice movement, in which educated community advocates and even affected community members have called attention to evidence of a higher pollution burden in minority and/or lower SES communities. The intention of this paper is to 1) examine the development and understanding of CRA, primarily by the U.S. Environmental Protection Agency; 2) to assess several states agencies and some EPA regional offices' interpretation of CRA, again based primarily on EPA guidance, and 3) to analyze how CRA might be refined in its implementation—giving some cues as to how the EPA may more effectively interact with communities interested in CRA.^

Anelastic strain recovery and elastic properties of ODP Leg 123 oceanic basalt samples

A knowledge of rock stress is fundamental for improving our understanding of oceanic crustal mechanisms and lithospheric dynamic processes. However, direct measurements of stress in the deep oceans, and in particular stress magnitudes, have proved to be technically difficult. Anelastic strain recovery measurements were conducted on 15 basalt core samples from Sites 765 and 766 during Leg 123. Three sets of experiments were performed: anelastic strain recovery monitoring, dynamic elastic property measurements, and thermal azimuthal anisotropy observations. In addition, a range of other tests and observations were recorded to characterize each of the samples. One common feature of the experimental results and observations is that apparently no consistent orientation trend exists, either between the different measurements on each core sample or between the same sets of measurements on the various core samples. However, some evidence of correspondence between velocity anisotropy and anelastic strain recovery exists, but this is not consistent for all the core samples investigated. Thermal azimuthal anisotropy observations, although showing no conclusive correlations with the other results, were of significant interest in that they clearly exhibited anisotropic behavior. The apparent reproducibility of this behavior may point toward the possibility of rocks that retain a "memory" of their stress history, which could be exploited to derive stress orientations from archived core. Anelastic strain recovery is a relatively new technique. Because use of the method has extended to a wider range of rock types, the literature has begun to include examples of rocks that contracted with time. Strong circumstantial evidence exists to suggest that core-sample contractions result from the slow diffusion of pore fluids from a preexisting microcrack structure that permits the rock to deflate at a greater rate than the expansion caused by anelastic strain recovery. Both expansions and contractions of the Leg 123 cores were observed. The basalt cores have clearly been intersected by an abundance of preexisting fractures, some of which pass right through the samples, but many are intercepted or terminate within the rock matrix. Thus, the behavior of the core samples will be influenced not only by the properties of the rock matrix between the fractures, but also by how these macro- and micro-scale fractures mutually interact. The strain-recovery curves recorded during Leg 123 for each of the 15 basalt core samples may reflect the result of two competing time dependent processes: anelastic strain recovery and pore pressure recovery. Were these the only two processes to influence the gauge responses, then one might expect that given the additional information required, established theoretical models might be used to determine consistent stress orientations and reliable stress magnitudes. However, superimposed upon these competing processes is their respective interaction with the preexisting fractures that intersect each core. Evidence from our experiments and observations suggests that these fractures have a dominating influence on the characteristics of the recovery curves and that their effects are complex.

More Is Not Always Better: An Experimental Individual-Level Validation of the Randomized Response Technique and the Crosswise Model

Social desirability and the fear of sanctions can deter survey respondents from responding truthfully to sensitive questions. Self-reports on norm breaking behavior such as shoplifting, non-voting, or tax evasion may therefore be subject to considerable misreporting. To mitigate such misreporting, various indirect techniques for asking sensitive questions, such as the randomized response technique (RRT), have been proposed in the literature. In our study, we evaluate the viability of several variants of the RRT, including the recently proposed crosswise-model RRT, by comparing respondents’ self-reports on cheating in dice games to actual cheating behavior, thereby distinguishing between false negatives (underreporting) and false positives (overreporting). The study has been implemented as an online survey on Amazon Mechanical Turk (N = 6,505). Our results indicate that the forced-response RRT and the unrelated-question RRT, as implemented in our survey, fail to reduce the level of misreporting compared to conventional direct questioning. For the crosswise-model RRT, we do observe a reduction of false negatives (that is, an increase in the proportion of cheaters who admit having cheated). At the same time, however, there is an increase in false positives (that is, an increase in non-cheaters who falsely admit having cheated). Overall, our findings suggest that none of the implemented sensitive questions techniques substantially outperforms direct questioning. Furthermore, our study demonstrates the importance of distinguishing false negatives and false positives when evaluating the validity of sensitive question techniques.

Influence of nutritional and management practices on productive performance and egg quality of brown egg-laying hens = Influencia de factores nutricionales y de manejo sobre la productividad y calidad del huevo en gallinas ponedoras rubias

El objetivo general de esta Tesis Doctoral fue estudiar la influencia de diversos factores nutricionales y de manejo sobre la productividad y la calidad del huevo en gallinas ponedoras comerciales rubias. Los factores estudiados fueron: 1) Cereal principal y tipo de grasa en la dieta; 2) Nivel de proteína bruta y grasa en la dieta; 3) Nivel energético de la dieta; 4) Peso vivo al inicio del período de puesta. En el experimento 1, la influencia del cereal principal en la dieta y el tipo de grasa suplementada en la dieta sobre los parámetros productivos y la calidad del huevo fue estudiado en 756 gallinas rubias de la estirpe Lohmann desde la sem 22 hasta las 54 de vida. El experimento se realizó mediante un diseño completamente al azar con 9 tratamientos ordenados factorialmente, con 3 cereales bases (maíz, trigo blando y cebada) y 3 tipos de grasa que variaban en su contenido en ácido linoléico (aceite de soja, oleína vegetal mezcla y manteca). Todas las dietas satisfacian las recomendaciones nutricionales para gallinas ponedoras rubias según el NRC (1994) y FEDNA (2008). La unidad experimental fue la jaula para todas las variables. Cada tratamiento fue replicado 4 veces, y la unidad experimental estuvo formada por 21 gallinas alojadas en grupos de 7. Las dietas fueron formuladas con un contenido nutritivo similar, excepto para el ácido linoléico, que varió en función del tipo de cereal y grasa utilizado. Así, dependiendo de la combinación de estos elementos el contenido de este ácido graso varió desde un 0.8% (dieta trigo-manteca) a un 3.4% (dieta maíz-aceite de soja). Este rango de ácido linoléico permitió estimar el nivel mínimo de este nutriente en el pienso que permite maximizar el peso del huevo. Los parámetros productivos y la calidad del huevo se controlaron cada 28 días y el peso de las aves se midió individualmente al inicio y al final del experimento con el objetivo de estudiar la variación en el peso vivo de los animales. No se observaron interacciones entre el tipo de cereal y grasa en la dieta para ninguna de las variables productivas estudiadas. Los tratamientos experimentales no afectaron a las principales variables productivas (porcentaje de puesta, peso del huevo y masa de huevo). Sin embargo, la ganancia de peso fue mayor en gallinas alimentadas con maíz o trigo que las gallinas alimentadas con cebada (243 vs. 238 vs. 202 g, respectivamente; P< 0.05). En el mismo sentido, las gallinas alimentadas con manteca obtuvieron una mayor ganancia de peso que las gallinas alimentadas con aceite de soja u oleína vegetal (251 vs. 221 vs. 210 g, respectivamente; P< 0.05). En cuanto a las variables estudiadas en relación con la calidad del huevo, ninguna de las variables estudiadas se vio afectada por el tratamiento experimental, salvo la pigmentación de la yema. Así, las gallinas alimentadas con maíz como cereal principal obtuvieron una mayor puntuación en relación con la escala de color que las gallinas alimentadas con trigo y con cebada (9.0 vs. 8.3 vs. 8.3, respectivamente; P< 0.001). La pigmentación de la yema también se vio afectada por el tipo de grasa en la dieta, así, las gallinas alimentadas con manteca obtuvieron una mayor puntuación de color en relación con la escala de color que las gallinas alimentadas con aceite de soja u oleína vegetal (8.9 vs. 8.5 vs. 8.2, respectivamente; P< 0.001). La influencia del contenido en ácido linoléico respecto al peso de huevo y masa de huevo fue mayor a medida que el contenido de dicho ácido graso se redujo en la dieta. Así, la influencia de la dieta en los radios peso de huevo/g linoléico ingerido y masa de huevo/g linoléico ingerido fue significativamente mayor a medida que el contenido en dicho ácido graso disminuyo en la dieta (P< 0.001). Los resultados del ensayo indican que las gallinas ponedoras rubias no necesitan más de un 1.0% de ácido linoléico en la dieta para maximizar la producción y el tamaño del huevo. Además, se pudo concluir que los 3 cereales y las 3 grasas utilizadas pueden sustituirse en la dieta sin ningún perjuicio productivo o referente a la calidad del huevo siempre que los requerimientos de los animales sean cubiertos. En el experimento 2, la influencia del nivel de proteína bruta y el contenido de grasa de la dieta sobre los parámetros productivos y la calidad del huevo fue estudiado en 672 gallinas ponedoras rubias de la estirpe Lohmann entre las sem 22 y 50 de vida. El experimento fue conducido mediante un diseño completamente al azar con 8 tratamientos ordenados factorialmente con 4 dietas y 2 pesos vivos distintos al inicio de puesta (1592 vs. 1860g). Tres de esas dietas diferían en el contenido de proteína bruta (16.5%, 17.5% y 18.5%) y tenían un contenido en grasa añadida de 1.8%. La cuarta dieta tenía el nivel proteico más elevado (18.5%) pero fue suplementada con 3.6% de grasa añadida en vez de 1.8%. Cada tratamiento fue replicado 4 veces y la unidad experimental consistió en 21 gallinas alojadas dentro de grupos de 7 animales en 3 jaulas contiguas. Todas las dietas fueron isocalóricas (2750 kcal EMAn/kg) y cubrieron las recomendaciones en aminoácidos para gallinas ponedoras rubias (Arg, Ile, Lys, Met, Thr, Trp, TSAA y Val) según el NRC (1994) y FEDNA (2008). Los efectos de los tratamientos sobre las variables productivas y la calidad de huevo fueron estudiados cada 28 días. La dieta no afecto a ninguna de las variables productivas estudiadas a lo largo del período productivo. Sin embargo, el peso inicial origino que las gallinas pesadas consumieran más (120.6 vs. 113.9 g; P< 0.001), obtuvieran un porcentaje de puesta mayor (92.5 vs. 89.8%; P< 0.01) y un peso del huevo mayor (64.9 vs. 62.4 g; P< 0.001) que las gallinas ligeras. El peso inicial de las gallinas no afecto al IC por kg de huevo ni a la mortalidad, sin embargo, la ganancia de peso fue mayor (289 vs. 233 g; P< 0.01) y el IC por docena de huevos fue mejor (1.52 vs. 1.57; P< 0.01) en las gallinas ligeras que en las gallinas pesadas. En cuanto a la calidad del huevo, la dieta no influyó sobre ninguna de las variables estudiadas. Los resultados del ensayo muestran que las gallinas ponedoras rubias, independientemente de su peso vivo al inicio de la puesta, no necesitan una cantidad de proteína bruta superior a 16.5% para maximizar la producción, asegurando que las dietas cubren los requerimientos en AA indispensables. Asimismo, se puedo concluir que las gallinas con un peso más elevado al inicio de puesta producen más masa de huevo que las gallinas con un peso más bajo debido a que las primeras producen más cantidad de huevos y más pesados. Sin embargo, ambos grupos de peso obtuvieron el mismo IC por kg de huevo y las gallinas más livianas en peso obtuvieron un mejor IC por docena de huevo que las pesadas. En el experimento 3 la influencia de la concentración energética sobre los parámetros productivos y la calidad del huevo fue estudiada en 520 gallinas ponedoras rubias de la estirpe Hy-Line en el período 24-59 sem de vida. Se utilizaron 8 tratamientos ordenados factorialmente con 4 dietas que variaron en el contenido energético (2650, 2750, 2850 y 2950 kcal EMAn/kg) y 2 pesos vivos distintos al inicio del período de puesta (1733 vs. 1606g). Cada tratamiento fue replicado 5 veces y la unidad experimental consistió en una jaula con 13 aves. Todas las dietas se diseñaron para que tuvieran una concentración nutritiva similar por unidad energética. Las variables productivas y de calidad de huevo se estudiaron mediante controles cada 28 días desde el inicio del experimento. No se observaron interacciones entre el nivel energético y el peso inicial del ave para ninguna de las variables estudiadas. Un incremento en la concentración energética de la dieta incrementó la producción de huevos (88.8 % vs. 91.2 % vs. 92.7 % vs. 90.5 %), masa de huevo (56.1 g/d vs. 58.1 g/d vs. 58.8 g/d vs. 58.1 g/d), y eficiencia energética (5.42 vs. 5.39 vs. 5.38 vs. 5.58 kcal EMA/g huevo) de forma lineal y cuadrática (P< 0.05) y afectó significativamente a la ganancia de peso (255 g vs. 300 g vs. 325 g vs. 359 g; P<0.05) . Sin embargo, un incremento en la concentración energética provocó un descenso lineal en el consumo de los animales (115 g vs. 114 g vs. 111 g vs. 110 g; P< 0.001) y un descenso lineal y cuadrático en el IC por kg de huevo (2.05 vs. 1.96 vs. 1.89 vs. 1.89; P< 0.01). En cuanto a la calidad del huevo, un incremento en el contenido energético de la dieta provocó una reducción en la calidad del albumen de forma lineal en forma de reducción de Unidades Haugh (88.4 vs. 87.8 vs. 86.3 vs. 84.7; P< 0.001), asimismo el incremento de energía redujo de forma lineal la proporción relativa de cáscara en el huevo (9.7 vs. 9.6 vs. 9.6 vs. 9.5; P< 0.001). Sin embargo, el incremento energético propició un incremento lineal en la pigmentación de la yema del huevo (7.4 vs. 7.4 vs. 7.6 vs. 7.9; P< 0.001). El peso vivo al inicio de la prueba afecto a las variables productivas y a la calidad del huevo. Así, los huevos procedentes de gallinas pesadas al inicio de puesta tuvieron una mayor proporción de yema (25.7 % vs. 25.3 %; P< 0.001) y menor de albumen (64.7 vs. 65.0; P< 0.01) y cáscara (9.5 vs. 9.6; P< 0.05) respecto de los huevos procedentes de gallinas ligeras. Consecuentemente, el ratio yema:albumen fue mayor (0.40 vs. 0.39; P< 0.001) para las gallinas pesadas. Según los resultados del experimento se pudo concluir que las actuales gallinas ponedoras rubias responden con incrementos en la producción y en la masa del huevo a incrementos en la concentración energética hasta un límite que se sitúa en 2850 kcal EMAn/kg. Asimismo, los resultados obtenidos entre los 2 grupos de peso al inicio de puesta demostraron que las gallinas pesadas al inicio de puesta tienen un mayor consumo y producen huevos más pesados, con el consecuente aumento de la masa del huevo respecto de gallinas más ligeras. Sin embargo, el IC por kg de huevo fue el mismo en ambos grupos de gallinas y el IC por docena de huevo fue mejor en las gallinas ligeras. Asimismo, la eficiencia energética fue mejor en las gallinas ligeras. Abstract The general aim of this PhD Thesis was to study the influence of different nutritional factors and management on the productivity and egg quality of comercial Brown laying hens. The factor studied were: 1) The effect of the main cereal and type of fat of the diet; 2) The effect of crude protein and fat content of the diet; 3) The effect of energy concentration of the diet; 4) The effect of initial body weight of the hens at the onset of lay period. In experiment 1, the influence of the main cereal and type of supplemental fat in the diet on productive performance and egg quality of the eggs was studied in 756 Lohmann brown-egg laying hens from 22 to 54 wk of age. The experiment was conducted as a completely randomized design with 9 treatments arranged factorially with 3 cereals (dented corn, soft wheat, and barley) and 3 types of fat (soy oil, acidulated vegetable soapstocks, and lard). Each treatment was replicated 4 times (21 hens per replicate). All diets were formulated according to NRC (1994) and FEDNA (2008) to have similar nutrient content except for linoleic acid that ranged from 0.8 (wheat-lard diet) to 3.4% (corn-soy bean oil) depending on the combination of cereal and fat source used. This approach will allow to estimate the minimum level of linoleic acid in the diets that maximizes egg weight. Productive performance and egg quality traits were recorded every 28 d and BW of the hens was measured individually at the beginning and at the end of the experiment. No significant interactions between main factors were detected for any of the variables studied. Egg production, egg weight, and egg mass were not affected by dietary treatment. Body weight gain was higher (243 vs. 238 vs. 202 g; P<0.05) for hens fed corn or wheat than for hens fed barley and also for hens fed lard than for hens fed soy oil or acidulated vegetable soapstocks (251 vs. 221 vs. 210 g; P< 0.05). Egg quality was not influenced by dietary treatment except for yolk color that was greater (9.0 vs. 8.3 vs. 8.3; P< 0.001) for hens fed corn than for hens fed wheat or barley and for hens fed lard than for hens fed soy oil or acidulated vegetable soapstocks (8.9 vs. 8.5 vs. 8.2, respectivamente; P< 0.001). The influence of linoleic acid on egg weight and egg mass was higher when the fatty acid was reduced in the diet. Thus, the influence of the diet in egg weight/g linoleic acid intake and egg mass/g linolec acid intake was higher when the amount of this fatty acid decreased in the diet (P< 0.001). It is concluded that brown egg laying hens do not need more than 1.0% of linoleic acid in the diet (1.16 g/hen/d) to maximize egg production and egg size. The 3 cereals and the 3 fat sources tested can replace each other in the diet provided that the linoleic acid requirements to maximize egg size are met. In experiment 2, the influence of CP and fat content of the diet on performance and egg quality traits was studied in 672 Lohmann brown egg-laying hens from 22 to 50 wk of age. The experiment was conducted as a completely randomized design with 8 treatments arranged factorially with 4 diets and 2 initial BW of the hens (1,592 vs. 1,860 g). Three of these diets differed in the CP content (16.5, 17.5, and 18.5%) and included 1.8% added fat. The fourth diet had also 18.5% CP but was supplemented with 3.6% fat instead of 1.8% fat. Each treatment was replicated 4 times and the experimental unit consisted of 21 hens allocated in groups of 7 in 3 adjacent cages. All diets were isocaloric (2,750 kcal AME/kg) and met the recommendations of brown egg-laying hens for digestible Arg, Ile, Lys, Met, Thr, Trp, TSAA, and Val. Productive performance and egg quality were recorded by replicate every 28-d. For the entire experimental period, diet did not affect any of the productive performance traits studied but the heavier hens had higher ADFI (120.6 vs. 113.9g; P< 0.001), egg production (92.5 vs. 89.8%; P< 0.01), and egg weight (64.9 vs. 62.4g; P< 0.001) than the lighter hens. Initial BW did not affect feed conversion per kilogram of eggs or hen mortality but BW gain was higher (289 vs. 233g; P< 0.01) and FCR per dozen of eggs was better (1.52 vs. 1.57; P< 0.01) for the lighter than for the heavier hens. None of the egg quality variables studied was affected by dietary treatment or initial BW of the hens. It is concluded that brown egg-laying hens, irrespective of their initial BW, do not need more than 16.5% CP to maximize egg production provided that the diet meet the requirements for key indispensable amino acids. Heavier hens produce more eggs that are larger than lighter hens but feed efficiency per kilogram of eggs is not affected. In experiment 3, the influence of AMEn concentration of the diet on productive performance and egg quality traits was studied in 520 Hy-Line brown egg-laying hens differing in initial BW from 24 to 59 wks of age. There were 8 treatments arranged factorially with 4 diets varying in energy content (2,650, 2,750, 2,850, and 2,950 kcal AMEn/kg) and 2 initial BW of the hens (1,733 vs. 1,606 g). Each treatment was replicated 5 times (13 hens per replicate) and all diets had similar nutrient content per unit of energy. No interactions between energy content of the diet and initial BW of the hens were detected for any trait. An increase in energy concentration of the diet increased (linear, P< 0.05; quadratic P< 0.05) egg production (88.8 % vs. 91.2 % vs. 92.7 % vs. 90.5 %), egg mass (56.1 g/d vs. 58.1 g/d vs. 58.8 g/d vs. 58.1 g/d), energy efficiency (5.42 vs. 5.39 vs. 5.38 vs. 5.58 kcal AMEn/g of egg), and BW gain (255 g vs. 300 g vs. 325 g vs. 359 g; P<0.05) but decreased ADFI (115 g vs. 114 g vs. 111 g vs. 110 g; P< linear, P< 0.001) and FCR per kg of eggs (2.05 vs. 1.96 vs. 1.89 vs. 1.89; linear, P< 0.01; quadratic P< 0.01). An increase in energy content of the diet reduced Haugh units (88.4 vs. 87.8 vs. 86.3 vs. 84.7; P< 0.01) and the proportion of shell in the egg (9.7 vs. 9.6 vs. 9.6 vs. 9.5; P< 0.001). Feed intake (114.6 vs. 111.1 g/hen per day), AMEn intake (321 vs. 311 kcal/hen per day), egg weight (64.2 vs. 63.0 g), and egg mass (58.5 vs. 57.0 g) were higher for the heavier than for the lighter hens (P<0.01) but FCR per kg of eggs and energy efficiency were not affected. Eggs from the heavier hens had higher proportion of yolk (25.7 % vs. 25.3 %; P< 0.001) and lower of albumen (64.7 vs. 65.0; P< 0.01) and shell (9.5 vs. 9.6; P< 0.05) than eggs from the lighter hens. Consequently, the yolk to albumen ratio was higher (0.40 vs. 0.39; P< 0.001) for the heavier hens. It is concluded that brown egg-laying hens respond with increases in egg production and egg mass, to increases in AMEn concentration of the diet up to 2,850 kcal/kg. Heavy hens had higher feed intake and produced heavier eggs and more egg mass than light hens. However, energy efficiency was better for the lighter hens.

Quick time planning using "s" curves and cost based durations.

ABSTRACT: The comparison of the different bids in the tender for a project, with the traditional contract system based on unit rates open to and re-measurement, requires analysis tools that are able to discriminate proposals having a similar overall economic impact, but that might show a very different behaviour during the execution of the works. RESUMEN: La estimación rápida de costes en fases iniciales del proyecto por métodos paramétricos y referencias estadísticas es un tema bien estudiado, divulgado y aplicado en el sector de la construcción. Sin embargo, existe poca literatura técnica sobre sistemas de predimensionado de tiempos, que permitan realizar rápidamente una planificación con un grado de aproximación razonable. Este texto reúne dos aspectos ya conocidos, pero hasta ahora independientes, y una aportación propia:  -La estimación del plazo final por referencias estadísticas (BCIS, 2000)  - La estimación del reparto del coste total a lo largo de la ejecución mediante curvas "S" (diversos autores)  La estimación de la duración de la ejecución de las actividades en función de su coste. El conjunto de estas tres técnicas, aplicadas a un proyecto, permite obtener una planificación con el suficiente grado de detalle y fiabilidad para tomar decisiones en fases iniciales del proyecto.

Risk of Leaching in Soils Amended by Compost and digestate from Municipal Solid Waste

New European directives have proposed the direct application of compost and digestate produced from municipal solid wastes as organic matter sources in agricultural soils. Therefore information about phosphorus leaching from these residues when they are applied to the soil is increasingly mportant. Leaching experiments were conducted to determine the P mobility in compost and digestate mixtures, supplying equivalent amounts to 100 kg P ha?1 to three different types of soils. The tests were performed in accordance with CEN/TS 14405:2004 analyzing the maximum dissolved reactive P and the kinetic rate in the leachate. P biowaste fractionation indicated that digestate has a higher level of available P than compost has. In contrast, P losses in leaching experiments with soil-compost mixtureswere higher than in soil-digestate mixtures. For bothwastes, therewas no correlation between disolved reactive P lost and the water soluble P.The interaction between soil and waste, the long experimentation time, and the volume of leachate obtained caused the waste?s wettability to become an influential parameter in P leaching behavior. The overall conclusion is that kinetic data analysis provides valuable information concerning the sorption mechanism that can be used for predicting the large-scale behavior of soil systems.

New systems of management to improve the health and welfare of rabbits in farm conditions = Nuevos sistemas de cría y manejo para mejorar la salud y bienestar de conejos de granja

El objetivo general de esta Tesis Doctoral fue evaluar nuevos sistemas de alojamiento y cría de conejos de granja, estudiando tanto parámetros comportamentales (experimento 1) como productivos y reproductivos (experimento 3). Además, se evaluaron diferentes técnicas de muestreo con el fin de optimizar el tiempo empleado para el estudio del comportamiento animal (experimento 2). En el experimento 1, se estudió el comportamiento de conejas alojadas en dos tipos de jaulas (TJ), convencionales vs. alternativas con una plataforma elevada, en distintos estados fisiológicos (EF), lactantes y gestantes. Se observó el comportamiento de 12 conejas reproductoras con grabaciones de una duración de 24 h continuas. Independientemente del EF y TJ, las conejas pasaron gran parte de su tiempo sobre el reposapatas (57,7 %, de media). Sin embargo, debido al uso de la plataforma (23,0% del tiempo, de media), las conejas lactantes permanecieron un 36,6 % menos de tiempo (P<0,001) sobre el reposapatas y las gestantes un 27,0% menos (P<0,001) sobre el enrejillado en jaulas alternativas que en convencionales. En las jaulas alternativas, las conejas podían adoptar la postura “levantada”, sin embargo ésta fue observada solamente en conejas gestantes una media de 4,6 veces al día. Las conejas bebieron con mas frecuencia en jaulas convencionales que en alternativas (24,6 vs 19,1 veces al día; P<0,05). Se observó una mayor duración y frecuencia del comportamiento “interactuando con compañeras” en conejas gestantes alojadas en jaulas convencionales (276 s/d y 4,6 veces/d; P<0,05). La frecuencia de “interactuando con gazapos” fue menor en jaulas alternativas que en convencionales (2,4 vs 8,6 veces al día; P<0,01). La hora del día afectó al comportamiento de las conejas, teniendo un comportamiento menos activo durante las horas centrales del día. Durante las horas de oscuridad, las conejas estuvieron más inquietas realizando comportamientos como ‘encabritarse’ o amamantar, coincidiendo éstos en el tiempo en el cual las conejas pasaron más tiempo en la plataforma. Las conejas utilizaron frecuentemente la plataforma, independientemente del estado fisiológico. En la fase de lactación, las conejas utilizaron la plataforma para huir de los intentos de mamar por parte de los gazapos cuando éstas no estaban receptivas. El uso de la plataforma puede dar lugar a problemas higiénicos debidos tanto por la acumulación de heces sobre ella como por la caída de heces y orina sobre los animales que están en la parte inferior. La ausencia de estereotipias por parte de las conejas tanto en jaulas alternativas como en convencionales no sugiere una falta de bienestar debida al sistema de alojamiento. En el experimento 2, se compararon distintos métodos de observación simplificada con respecto un método de referencia usando grabaciones continuas de 24 h para la evaluación del comportamiento de conejas en distintos estados fisiológicos (gestantes y lactantes) alojadas en dos tipos de jaulas (convencionales y alternativas). Se analizaron un total de 576 h de grabaciones continuas de 24 h en 12 conejas reproductoras al final del periodo de lactación y en las mismas conejas después del destete. Los comportamientos observados se clasificaron en tres categorías independientes (localización en la jaula, postura y comportamientos funcionales). Se utilizaron grabaciones continuas de 24 h como método de referencia para validar otros cuatro métodos de observación simplificados, utilizando grabaciones de distinta duración y frecuencia a lo largo del día. Métodos regulares: corto y largo con 2.4 y 8 h de observación respectivamente, y métodos irregulares: corto y largo con 6 y 8 h de observación, respectivamente. Como resultado, se observó que independientemente del sistema de alojamiento, el mejor método para reducir el tiempo de observación necesario para evaluar el comportamiento de conejas reproductoras depende del tipo de variable a estudiar y del estado fisiológico de las conejas. En gestantes, los métodos irregulares no fueron adecuados para estimar comportamientos de larga duración tales como tumbada, sentada, descansando y acicalándose. Sin embargo, en ambos estados fisiológicos, los métodos regulares fueron precisos para los comportamientos de los grupos localización y postura y para comportamientos funcionales de larga duración. Por otro lado, los coeficientes de variación de los comportamientos poco frecuentes realizados principalmente durante el periodo de oscuridad fueron muy altos, y el método irregular largo obtuvo los menores errores de estimación para éstos comportamientos. En el experimento 3, se estudió el efecto de un uso combinado de lactaciones largas (hasta 46 días) con jaulas alternativas sobre los parámetros productivos y reproductivos de 104 conejas y sus camadas durante cinco ciclos reproductivos. La mitad de las conejas fueron alojadas en jaulas polivalentes convencionales (39 cm x 100 cm x 30 cm) y la otra mitad en jaulas polivalentes alternativas (39 cm x 100 cm x 60 cm), con una plataforma elevada. Dentro de cada grupo de alojamiento, la mitad de las conejas se destetaron a 32 días y la otra mitad a 46 días tras el parto. Las lactaciones más largas afectaron negativamente al peso (P<0,001), contenido en grasa y energía (P<0,05) de las conejas al final del periodo de lactación, pero éste efecto disminuyó con el número de partos. La fertilidad, prolificidad y la mortalidad de las conejas no fue afectada por la duración de la lactación. El destete tardío dio lugar a un mayor tamaño y peso de la camada al final del periodo de crecimiento (8,9 y 11,3 %, respectivamente) y a un menor índice de conversión por jaula durante el todo el periodo experimental (13,5 %) con respecto al destete convencional (P<0,001). Éstos resultados fueron paralelos a la menor mortalidad global (12,6 vs 17,6 %; P<0,05) observada en gazapos con destete tardío. Las diferencias en los parámetros productivos con las distintas edades al destete sólo fueron observadas en los ciclos con peor estado sanitario (tercer y quinto ciclo), en los cuales el destete tardío redujo la mortalidad. El tipo de jaula no afectó al peso de la coneja, condición corporal, mortalidad, fertilidad ni tamaño de camada durante los cinco primeros ciclos reproductivos. Sin embargo, el peso de la camada y el índice de conversión a los 21 días de edad fueron 4,2% mayor (P<0,001) y 5,0% menor (P<0,005) en animales alojados en jaulas alternativas que en jaulas convencionales. A día 59 las jaulas alternativas dieron lugar a camadas más pesadas (P<0,01); sin embargo, éste efecto fue influenciado por la densidad alcanzada en cada ciclo, ya que cuando la densidad de los animales fue menor que 40kg/m2 (tercer y quinto ciclo), el efecto del tipo de jaula sobre el peso de la camada no fue significativo. De los resultados obtenidos se puede concluir que el uso combinado de lactaciones más largas y jaulas con mayor superficie disponible con una plataforma elevada podría ser una alternativa para mejorar el bienestar animal en determinadas situaciones productivas. ABSTRACT The general aim of this PhD Thesis was to evaluate new housing and husbandry systems of farmed rabbits, studying behavioral (experiment 1), productive and reproductive (experiment 3) parameters. Moreover, different sampling techniques were evaluated in order to optimize the assessment of rabbit behaviour (experiment 2). In experiment 1, the behaviour of rabbit does housed in two different types of cage (TC), conventional vs. alternative with an elevated platform, at different physiological stages (PS), lactation and gestation was to study. Behavioural observations were carried out on 12 commercial rabbit does using continuous 24 hour video recording. Independently of PS and TC, rabbit does spent most of their time on foot mats (57.7 %, as average). However, due to the use of platforms (23.0% of time, as average), lactating does spent 36.6% less time (P<0.001) on foot mats and gestating does spent 27.0% less (P<0.001) time on wire mesh in alternative cages than in conventional cages. Alternative cages allowed for standing posture but this behaviour was only observed in gestating does (4.6 times a day, as average). Frequency of drinking was higher in conventional than in alternative cages (24.6 vs. 19.1 times a day; P<0.05). Gestating does housed in conventional cages reached the highest duration and frequency of interacting with neighbours (276 s/d and 4.6 times/d; P<0.05). The frequency of interacting with kits was lower in alternative than in conventional cages (2.4 vs. 8.6 times a day; P<0.01). Does’ behaviour was influenced by hour of day, being less active at the midday hours. During dark hours rabbit does more frequently performed restless behaviour such as hyperactivity or nursing, matching the time at which rabbit does spent more time on the platform. The platform was frequently used by rabbit does, independent of their physiological state, and during late lactation phase, when mothers were not receptive to nursing, does housed in alternative cages used the platform as a mean to flee from kids trying to suckle. The use of the platform might lead to hygienic problems due to retained faeces on the platform and faeces and urine falling onto animals located in the lower part of the cage. Stereotypies were not observed in any housing system, therefore conventional cages do not suggest lack of animal welfare. In experiment 2, it was compared the results of different simplified sampling methods of behavioural data with respect to reference records of 24-h in order to assess rabbit does behaviours at different physiological stages (gestation and lactation) in animals housed in two types of cages (conventional and alternative). A total of 576 h of continuous video of 12 rabbit does at the end of lactation and on the same females after weaning were analysed. The behavioural observations were studied using three independent categories of classification (location in the cage, posture and functional behaviours). Continuous behavioural recordings of 24 h were considered as the reference method to validate another 4 sampling methods of data collection by aggregated video recordings of different frequency and duration (regular short and long methods with 2.4 and 8 h of observation respectively, and irregular short and long methods with 6 and 8 h of observation, respectively). The current results showed that, independently of housing system, the best method to reduce the total observation time required to assess rabbit does behaviour depends on the trait studied and physiological stage of does. In gestating does, irregular methods were not suitable to estimate behaviours of long duration such as lying, sitting, resting and grooming. However, in both physiological stages, regular methods were accurate for location behaviours, postures and functional behaviours of long duration. Instead, for the study of infrequent behaviours performed mainly during dark period, where coefficients of variation were high, the irregular long method led to the lowest mean estimation errors. In experiment 3, the effects of the combined use of long lactation periods (46 days) with alternative cages on the reproductive and growth performance of 104 rabbit does and their litters during five consecutive reproductive cycles were studied. Half of does were housed in conventional polyvalent cages (39 cm x 100 cm x 30 cm) and the other half in alternative polyvalent cages (39 cm x 100 cm x 60 cm), with a raised platform. Half of the rabbit does in each type of cages were weaned at 32 and the other half at 46 days after parturition. Longer lactations affected negatively to body weight (P<0.001), fat and energy content (P<0.05) of rabbit does at the end of the lactation period, but this effect decreased with the number of parturitions. Fertility, prolificacy and doe mortality were not affected by lactation length. Late weaning led to higher litter size (by 8.9 %) and litter weight (by 11.3 %) at the end of growing period and lower feed conversion ratio per cage during the overall experimental period (13.5 %) than standard weaning (P<0.001). These results were parallels to a lower mortality (12.6 vs 17.6 %; P<0.05) of young rabbit weaned later during the overall experimental period. Differences in performances at different weaning ages were only observed during cycles with worst health status (third and fifth cycles) in which late weaning decreased mortality. Type of cage did not affect doe body weight and body condition, mortality, fertility, prolificacy and litter size during the five firsts reproductive cycles. Nevertheless, at day 21 litter weight and feed conversion ratio were 4.2 % higher (P<0.001) and 5.0 % lower (P<0.005) in animals housed in alternative than in conventional cages. Alternative cages also led to heavier litters at 59 days (P<0.01); however, this effect was influenced by density reached in each cycle, as when the density of animals was lower than 40 kg/m2 (cycles three and five), the difference of litter weight between alternative and conventional cages was not significant. From the results obtained it can be concluded that the combined use of longer lactations and cages with higher available surface with a raised platform could be an alternative to improve animal welfare in some productive situations.

The small GTP-binding protein Rho links G protein-coupled receptors and Gα12 to the serum response element and to cellular transformation

Receptors coupled to heterotrimeric G proteins can effectively stimulate growth promoting pathways in a large variety of cell types, and if persistently activated, these receptors can also behave as dominant-acting oncoproteins. Consistently, activating mutations for G proteins of the Gαs and Gαi2 families were found in human tumors; and members of the Gαq and Gα12 families are fully transforming when expressed in murine fibroblasts. In an effort aimed to elucidate the molecular events involved in proliferative signaling through heterotrimeric G proteins we have focused recently on gene expression regulation. Using NIH 3T3 fibroblasts expressing m1 muscarinic acetylcholine receptors as a model system, we have observed that activation of this transforming G protein-coupled receptors induces the rapid expression of a variety of early responsive genes, including the c-fos protooncogene. One of the c-fos promoter elements, the serum response element (SRE), plays a central regulatory role, and activation of SRE-dependent transcription has been found to be regulated by several proteins, including the serum response factor and the ternary complex factor. With the aid of reporter plasmids for gene expression, we observed here that stimulation of m1 muscarinic acetylcholine receptors potently induced SRE-driven reporter gene activity in NIH 3T3 cells. In these cells, only the Gα12 family of heterotrimeric G protein α subunits strongly induced the SRE, while Gβ1γ2 dimers activated SRE to a more limited extent. Furthermore, our study provides strong evidence that m1, Gα12 and the small GTP-binding protein RhoA are components of a novel signal transduction pathway that leads to the ternary complex factor-independent transcriptional activation of the SRE and to cellular transformation.

Identification of a member of the interferon regulatory factor family that binds to the interferon-stimulated response element and activates expression of interferon-induced genes.

A family of interferon (IFN) regulatory factors (IRFs) have been shown to play a role in transcription of IFN genes as well as IFN-stimulated genes. We report the identification of a member of the IRF family which we have named IRF-3. The IRF-3 gene is present in a single copy in human genomic DNA. It is expressed constitutively in a variety of tissues and no increase in the relative steady-state levels of IRF-3 mRNA was observed in virus-infected or IFN-treated cells. The IRF-3 gene encodes a 50-kDa protein that binds specifically to the IFN-stimulated response element (ISRE) but not to the IRF-1 binding site PRD-I. Overexpression of IRF-3 stimulates expression of the IFN-stimulated gene 15 (ISG15) promoter, an ISRE-containing promoter. The murine IFNA4 promoter, which can be induced by IRF-1 or viral infection, is not induced by IRF-3. Expression of IRF-3 as a Gal4 fusion protein does not activate expression of a chloramphenicol acetyltransferase reporter gene containing repeats of the Gal4 binding sites, indicating that this protein does not contain the transcription transactivation domain. The high amino acid homology between IRF-3 and ISG factor 3 gamma polypeptide (ISGF3 gamma) and their similar binding properties indicate that, like ISGF3 gamma, IRF-3 may activate transcription by complex formation with other transcriptional factors, possibly members of the Stat family. Identification of this ISRE-binding protein may help us to understand the specificity in the various Stat pathways.