Diseño del equipamiento de un estudio de grabación digital

El presente proyecto describe la instalación de audio de un estudio de grabación digital musical. La finalidad de este proyecto es puramente educativa, afianzando conceptos que se han contemplado durante la carrera. La instalación tiene carácter ficticio, por lo que no tiene implementación real. Aun así, se ha intentado desarrollar con carácter profesional. El proyecto se ha dividido en varias fases de trabajo. Primeramente, se procedió a la búsqueda de información relativa a estudios de grabación, atendiendo principalmente a sus configuraciones. Paralelamente, se buscó información sobre los principales equipos dentro de un estudio de grabación y realizando un pequeño estudio de mercado. Posteriormente, se ha procedido a la elección de la configuración del equipamiento del estudio, atendiendo a las ventajas e inconvenientes de cada tipo de configuración. La tercera fase, corresponde a la elección de los equipos. Siguiendo la cadena de audio, se ha ido analizando la necesidad de cada uno de ellos. Seguidamente, se ha realizado una comparación de diferentes equipos que componen cada bloque de elección, y finalmente la selección del más apropiado junto con su justificación. En la última fase se ha realizado la interconexión de todos los equipos atendiendo a la configuración elegida en la segunda fase. Para ello, se ha llevado a cabo la implementación de una serie de tablas escritas, donde se especifica cada tipo de conexión. El proyecto ha terminado con una presentación del presupuesto, dividido en varios aparatados, y el desarrollo de las conclusiones. En ellas, se ha analizado tanto los objetivos propuestos al principio del proyecto como una valoración personal del proyecto en general. ABSTRACT. This project describes the audio installation of a digital music recording studio. The purpose of this project is purely educational, strengthening concepts that have been laid during college. The installation is fictitious and has not been implemented in a real situation. Nevertheless, it has been developed with a professional character. This project has been divided in various phases. Firstly, I proceeded to search information related to recording studios, focusing specially on their configurations. Simultaneously, I looked for information about the main digital equipment of a recording studio and performed a brief market research. Secondly, I selected the studio equipment configuration, taking care of the advantages and disadvantages of each type of configuration. The third phase corresponds to the choice of the equipment. Following the audio chain, I analyzed the need for each of them. Then, I compared the different equipment that compose each of the choice blocks and finally opt for the most appropriate with its justification. In the last phase, I interconnected all the equipment according to the chosen configuration of the second phase. For this, I implemented a series of written tables, where I specified each connection type. The Project ends with a presentation of the budget, divided into several sections, followed by the conclusion in which I analyze both the objectives of the project and my personal valuation.

Utilización de ingredientes tradicionales y nuevos productos derivados del haba de soja en piensos de lechones blancos e ibéricos: efectos sobre la fisiología digestiva y la productividad

Los objetivos principales de esta Tesis Doctoral fueron estudiar en 4 ensayos los efectos a) del procesado del maíz y la inclusión en los piensos de ingredientes de alta calidad como harina de pescado o fuentes de lactosa en lechones blancos b) inclusión en el pienso de diferentes productos derivados del haba de soja, con diferente contenido de proteína bruta (PB), tamaño de partícula y origen en lechones blancos e ibéricos y c) inclusión en el pienso de lechones ibéricos de ingredientes de alta calidad; forma de presentación del pienso y la duración del suministro del pienso prestárter sobre los parámetros productivos, la digestibilidad de los nutrientes, y las características morfológicas de la mucosa digestiva en lechones blancos e ibéricos recién destetados. En el experimento 1, los efectos de la complejidad del pienso prestárter sobre los parámetros productivos y la digestibilidad total aparente (TTAD) de los nutrientes fueron estudiados en lechones blancos recién destetados. Se utilizaron 10 tratamientos experimentales como resultado de 5 piensos prestárter (21 a 41 d de edad) y 2 piensos estárter (42 a 62 d de edad). Los piensos prestárter consistieron en un control negativo que incluía 40% de maíz crudo, 4% de harina de pescado y 7% de lactosa, un control positivo que incluía 40% de maíz cocido, 10% de harina de pescado, y 14% de lactosa, y 3 piensos adicionales con similares ingredientes que el pienso control positivo pero en los que a) 40% de maíz cocido fue sustituido por el mismo porcentaje de maíz crudo, b) se redujo el nivel de harina de pescado del 10 al 4%, y c) se redujo el nivel de lactosa del 14 al 7%. Cada tratamiento se replicó 6 veces (6 lechones/departamento). De 42 a 62 d de edad, la mitad de cada uno de los 5 piensos prestárter recibió un pienso estándar compuesto por harina de soja- maíz crudo y manteca y la otra mitad un pienso con similar perfil nutricional pero incluyendo un 20% de maíz cocido, 5% de harina de pescado, 1.3% de lactosa, 2% de concentrado de proteína de soja obtenido por fermentación y 1% de aceite de soja en lugar de harina de soja, maíz sin procesar y manteca. La complejidad del pienso no afectó a los parámetros productivos en ninguno de los periodos estudiados, pero el índice de diarreas durante la fase prestárter fue mayor en los lechones que recibieron el pienso control negativo que en los alimentados con cualquiera de los otros piensos (P<0.05). A los 30 días de edad (piensos prestárter), la digestibilidad de la materia orgánica (MO) y de la energía bruta (EB) fue menor (P<0.001) en los lechones que consumieron el pienso control negativo que en los lechones que consumieron cualquiera de los otros piensos. Sin embrago, la digestibilidad fecal de la PB no fue afectada. A los 50 días de edad (piensos estárter), la digestibilidad de los nutrientes fue similar en ambos piesnsos. Se concluye que la utilización de niveles elevados de ingredientes de alta calidad en los piensos no mejora los parámetros productivos de los lechones blancos en ninguno de los períodos estudiados. De 21 a 41 días de edad, el índice de diarreas se redujo y la digestibilidad de los nutrientes aumentó con la utilización de piensos de mayor calidad. Por lo tanto, la utilización de piensos con niveles elevados de ingredientes de calidad para reducir problemas digestivos y por lo tanto, mejorar los parámetros productivos podría estar justificada en algunos casos. En el experimento 2, se estudiaron los efectos de la inclusión en el pienso de harina de soja con diferente contenido de PB (44 vs. 49 % PB), la micronización de la harina de soja de alta proteína (AP-HS; 49% PB) y la utilización de concentrado de proteína de soja (CPS; 65% PB) sobre los parámetros productivos y la TTAD de los nutrientes en lechones blancos recién destetados de 28 a 63 días de edad. De 28 a 49 días de edad (fase I), hubo un pienso control positivo con un 10% de CPS, un pienso control negativo con 14.8% de harina de soja estándar (R-HS; 44% de PB) y otros 4 piensos que incluían 13.3% de AP-HS de origen Americano (USA) o Argentino (ARG) y molidas groseramente (980 μm) o micronizadas (80 μm). Cada tratamiento se replicó 8 veces (6 lechones/departamento). De 49 a 63 días de edad (fase II), todos los lechones recibieron un pienso comercial común en forma de harina. En el global de la fase I, el tratamiento experimental no afectó a ninguno de los parámetros productivos estudiados. Sin embargo, de 28 a 35 días de edad, los lechones alimentados con AP-HS micronizadas tuvieron un mejor índice de conversión (IC; 1.11 vs. 0.98; P<0.05) que los alimentados con AP-HS molidas groseramente. También, de 35 a 42 días de edad, los lechones que recibieron el pienso con AP-HS micronizada tendieron (P=0.08) a consumir más pienso que los lechones que consumieron el pienso con AP-HS molida. Durante la fase II (49 a 63 días de edad), cuando todos los lechones recibieron un pienso común, no se observaron diferencias en productividad de los lechones debido al tratamiento previo. En general, la digestibilidad de los nutrientes a los 35 días de edad fue mayor para los lechones que consumieron CPS que para los lechones que consumieron R-HS con los lechones que consumieron AP-HS en una posición intermedia. La digestibilidad de la PB fue mayor (P≤0.01) para el pienso que contenía CPS que para el promedio de los 5 tratamientos en base a HS. También, la digestibilidad de la MO y de la materia seca (MS) fue mayor para el pienso que contenía AP-HS micronizada o molida groseramente que para el pienso que contenía R-HS. La micronización de la AP-HS no tuvo efecto alguno sobre la digestibilidad de los nutrientes. Se concluye que cuando el CPS sustituye en el pienso a R-HS, la digestibilidad de la PB aumenta pero no tiene efecto alguno sobre los parámetros productivos. La utilización de AP-HS en sustitución de R-HS en el pienso mejora la digestibilidad de los nutrientes pero no afecta a los parámetros productivos. La utilización de harina de soja micronizada en los piensos mejora la eficiencia alimenticia durante la primera semana post-destete pero no tiene efecto alguno sobre la digestibilidad de los nutrientes. En general, la inclusión de productos derivados del haba de soja con un alto valor añadido (CPS o AP-HS) en el pienso presenta pocas ventajas en términos productivos al uso de AP-HS en lechones blancos recién destetados. En el experimento 3, se estudiaron los mismos productos de soja y piensos similares al experimento 2 en lechones ibéricos recién destetados. Además de los parámetros productivos y la TTAD de los nutrientes, en este ensayo se estudió también la digestibilidad ileal aparente (AID) de los nutrientes, así como las características histológicas y morfometría de la mucosa ileal. Cada uno de los 6 tratamientos fue replicado 6 veces (6 lechones/departamento). De 30 a 51 días de edad la fuente de harina de soja no afectó a los parámetros productivos, pero el índice de diarreas fue mayor (P<0.001) y la TTAD y AID de los nutrientes menor en los lechones alimentados con R-HS que en los alimentados con CPS o AP-HS. Sin embargo, no se encontró ninguna diferencia para éstos parámetros entre los piensos que contenían AP-HS y CPS. La TTAD de la MO (P=0.07) y de la EB (P=0.05) tendieron a ser mayores en los piensos basados en AP-HS micronizada que en los basados en AP-HS molida. La TTAD de la EB tendió (P<0.05) a ser mayor para la AP-HS de origen USA que para la AP-HS de origen ARG. Los lechones que consumieron R-HS presentaron villi de menor longitud (P<0.01) que los lechones que consumieron AP-HS o CPS, pero no se observaron diferencias en el caso de los lechones que recibieron los piensos que contenían AP-HS o CPS. Se concluye que la inclusión de AP-HS o CPS en el pienso en sustitución de R-HS reduce el índice de diarreas y mejora la digestibilidad de los nutrientes y las características morfológicas del íleon sin afectar a los parámetros productivos. La utilización de piensos basados en productos derivados del haba de soja con mayor valor añadido (CPS o AP-HS) en sustitución de la R-HS, mejora la TTAD de todos los nutrientes y reduce el índice de diarreas si llegar afectar a los parámetros productivos. En el experimento 4 se estudiaron los efectos del contenido de PB y la complejidad del pienso, la presentación física y la duración del suministro del pienso prestárter sobre los parámetros productivos y la TTAD de los nutrientes en lechones ibéricos recién destetados de 28 a 63 días de edad. Hubo 12 tratamientos experimentales con 2 tipos de pienso (AC; calidad alta y BC: calidad media), 2 presentaciones del pienso (gránulo y harina) y 3 duraciones de suministro del pienso prestárter (7, 14 y 21 días). Desde los 7, 14 y 21 días de experimento (dependiendo del tratamiento), hasta los 35 días, todos los lechones recibieron un pienso comercial en forma de harina. Cada uno de los tratamientos fue replicado 3 veces (6 lechones/departamento). En el global del experimento, la ganancia media diaria (GMD; P<0.05) y el consumo medio diario (CMD; P<0.01) fue menor en los lechones que recibieron el pienso AC que para los que recibieron el pienso de BC, si bien el IC no se vio afectado. La granulación del pienso prestárter no afectó a los crecimientos pero mejoró la eficiencia alimenticia. La utilización del pienso prestárter de 0 a 21 días de prueba mejoró el IC (P<0.05), pero redujo la GMD (P<0.01) en comparación con la utilización de éste pienso solo durante 7 o 14 días. El índice de diarreas tendió a ser mayor (P=0.06) en los lechones alimentados con los piensos AC que en los alimentados con los piensos BC. Asimismo, el índice de diarreas fue superior en los lechones que recibieron el pienso en gránulo que los que los recibieron en harina (P<0.001). Además, el índice de diarreas fue superior en los lechones que recibieron el pienso prestárter durante 14 o 21 días que en los que lo recibieron solo durante 7 días (P<0.01). De 28 a 49 días de edad, la GMD y el IC no se vieron afectados por la complejidad del pienso, pero la presentación en gránulo o el aumento en la duración de suministro del pienso prestárter mejoró el IC (P<0.01). También, en este periodo el índice de diarreas fue mayor en lechones alimentados con piensos granulados que aquellos alimentados con piensos en harina. Asimismo, fue superior para los lechones alimentados con el pienso prestárter durante 14 o 21 días que para los que recibieron éste pienso solo durante 7 días (P<0.01). De 49 a 63 días de edad, los lechones que previamente habían recibido piensos BC crecieron más que los que recibieron piensos AC (P<0.001). Asimismo, los lechones que recibieron el pienso prestárter durante 21 días comieron (P< 0.001) y crecieron menos (P<0.05) presentando una peor eficacia alimenticia (P<0.05) que los lechones que lo recibieron solo durante 7 14 días. La digestibilidad de la MO fue mayor en los lechones alimentados con los piensos AC que en los alimentados con piensos BC (P<0.05). La granulación del pienso mejoró la digestibilidad de los principales nutrientes. Los piensos prestárter AC mejoraron la digestibilidad de los nutrientes pero no la eficiencia alimenticia en lechones ibéricos de 28 a 63 días de edad. La granulación del pienso mejoró la eficiencia alimenticia. El aumento del suministro del pienso prestárter de 7 a 21 días mejoró la eficiencia alimenticia pero redujo la GMD. Por lo tanto, la utilización de piensos granulados de alta calidad durante el periodo prestárter es recomendable en lechones ibéricos, pero solo durante la primera semana post-destete. ABSTRACT The main objectives of this PhD Thesis were to study the effects of a) heat processing (HP) of corn and inclusion of high quality ingredients of animal origin such as fish meal (FM) and dried milk products in the diet, b) inclusion of different soy products varying in crude protein (CP) content, particle size, and origin of the beans in diets for conventional white and Iberian weanling pigs, and c) effects of ingredient quality, feed form, and duration of supply of the phase I diets on growth performance, nutrient digestibility, and intestinal morphology of weanling pigs. In experiment 1, the effect of diet complexity on total tract apparent digestibility (TTAD) and growth performance was studied in piglets from 21 to 62 d of age. There were 10 experimental treatments which resulted from the combination of 5 phase I (21 to 41 d of age) and 2 phase II (42 to 62 d of age) diets. The 5 phase I diets consisted of a negative control diet that contained 40 % raw corn, 4% FM, and 7% lactose (LAC); a positive control diet that contained 40 % HP corn, 10% FM, and 14% LAC, and 3 extra diets that used similar ingredients to those of the positive control diet but in which a) 40% of HP corn was substituted by raw corn, b) 4% FM rather than 10% FM, and c) 7% LAC instead of 14% LAC were included in the diet. Each treatment was replicated 6 times (6 pigs per pen). From 42 to 62 d of age, half of the pens of each of the 5 phase I treatments received a standard soybean meal (SBM)–native corn–lard diet wheras the other half received a diet with similar nutrient profile but that included 20% HP corn, 5% FM, 1.3% lactosa, 2% fermented soy protein concentrate, and 1% soybean oil in substitution of variables amounts of non-processed corn, SBM, and lard. Dietary treatment did not affect piglet performance at any age, but the incidence of post-weaning diarrhea (PWD) was higher during phase I in piglets fed the negative control diet than in piglets fed any of the other diets (P<0.05). At 30 d of age (phase I diets), the TTAD of organic matter (OM) and gross energy (GE) was lower (P<0.001) in pigs fed the negative control diet than in pigs fed the other diets but CP digestibility was not affected. At 50 d of age (phase II diets), dietary treatment did not affect TTAD of any dietary component. It is concluded that the use of high quality ingredients at high levels in the diet did not improve growth performance of piglets at any age. From 21 to 41 d of age, PWD was reduced and nutrient digestibility was increased in pigs fed the more complex diets. Consequently, the inclusion of high levels of high quality ingredients in piglet diets to maximize growth performance might not be justified under all circumstances In experiment 2, the effect of CP content (44 vs. 49 % CP) of SBM, micronization (fine grinding) of the high CP SBM (HP-SBM; 49% CP), and soy protein concentrate (SPC; 65% CP) on TTAD and growth performance was studied in conventional white piglets from 28 to 63 d of age. From 28 to 49 d of age (phase I), there was a positive control diet that included 6.5% CP from SPC and a negative control diet that supplied the same amount of CP as regular SBM (R-SBM; 44% CP) of Argentina (ARG) origin. The other 4 diets included the same amount of dietary CP from 2 sources of HP-SBM (USA or ARG origin), either ground (990 μm) or micronized (60 μm). Each treatment was replicated 8 times (6 pigs per pen). From 49 to 63 d of age (phase II), all pigs were fed a common commercial starter diet. For the entire phase I, type of soy product included in the diet did not affect growth performance of the pigs. However, from 28 to 35 d of age pigs fed the micronized HP-SBM had better feed conversion ratio (FCR; 0.90 vs. 1.01; P<0.05) than pigs fed the ground HP-SBM. Also, from 35 to 42 d of age, average daily feed intake (ADFI) tended to be higher (P=0.08) for pigs fed the micronized HP-SBM than for pigs fed the ground HP-SBM. During phase II, when all the pigs received the same diet, no differences among treatments were observed. In general, the TTAD of nutrients at 35 d of age was higher for the SPC than for the R-SBM diet with the HP-SBM diets being intermediate. The TTAD of CP was higher (83.8% vs. 81.9%; P≤0.01) for the SPC diet than for the average of 5 SBM containing diets. Also, the digestibility of OM and dry matter (DM) was higher (P<0.01) for the HP-SBM, either ground or micronized, than for the R-SBM diet. Micronization of the HP-SBM did not affect nutrient digestibility. It is concluded that when R-SBM was substituted by SPC, CP digestibility was improved but no effects on growth performance were observed. The use of HP-SBM in substitution of R-SBM in the diet improved nutrient digestibility but did not affect piglet performance. The inclusion of micronized HP-SBM in the diet improved FCR during the first week post-weaning but did not affect TTAD of nutrients. Therefore, the inclusion of added value soy products (SPC or micronized SBM) in the diet presents little advantage in terms of growth performance over the use of HP-SBM in pigs weaned at 28 d of age. In experiment 3, the effects of the same sources of soy protein used in experiment 2 on TTAD and growth performance of crossbreed Iberian pigs from 30 to 61 d of age were studied. In addition, the apparent ileal digestibility (AID) of nutrients and mucosa ileum morphology were also determined. Dietary treatment did not affect growth performance of the pigs at any age but from 30 to 51 d of age (phase I diets), PWD was higher (P<0.001) and the TTAD and AID of all nutrients were lower for pigs fed the R-SBM diet than for pigs fed the HP-SBM or the SPC diets. However, no differences between the HP-SBM and the SPC containing diets were detected for digestibility of any dietary component. The TTAD of OM (P=0.07) and GE (P=0.05) tended to be higher for the micronized HP-SBM than for the ground HP-SBM and that of GE was higher (P<0.05) for the USA meal than for the ARG meal. Pigs fed R-SBM had lower villus height (P<0.01) than pigs fed HP-SBM or SPC but no differences in ileal mucosal morphology were detected between SPC and HP-SBM containing diets. It is concluded that feeding the HP-SBM or SPC in substitution of R-SBM reduced PWD and improved nutrient digestibility and ileal morphology in piglets as compared with feeding the R-SBM, but had no effect on growth performance. The inclusion in the diet of added value soy products (micronized SBM or SPC) in substitution of the R-SBM increased the TTAD of all nutrients and reduced PWD but had no advantage in terms of growth performance over the use of ground HP-SBM. In experiment 4, the effect of CP content and ingredient complexity, feed form, and duration of feeding of the phase I diets on growth performance and TTAD of nutrients were studied in Iberian pigs from 28 to 63 d of age. There were 12 dietary treatments with 2 type of feeds (HQ; higher quality and LQ; medium quality), 2 feed forms (pellets vs. mash), and 3 durations of supply (7, 14, and 21 d) of the phase I diets. From d 7, 14, or 21 (depending on treatment) to d 35 of experiment, all pigs received a common diet in mash form. Each treatment was replicated 3 times (6 pigs/pen). For the entire experiment, average daily gain (ADG; P<0.05) and ADFI (P<0.01) were lower with the HQ than with the LQ phase I diets but FCR was not affected. Pelleting of the phase I diets did not affect ADG but improved FCR (P<0.01). Feeding the phase I diets from d 0 to 21 improved FCR (P<0.05) but decreased ADG (P<0.01) as compared with 7 or 14 d of feeding. Post-weaning diarrhea tended to be higher (P=0.06) for pigs fed the HQ diets than for pigs fed the LQ diets and for pigs fed pellets than for pigs fed mash (P<0.001). Also, PWD was higher for pigs fed the phase I diet for 14 or 21 d than for pigs fed this diet for 7 d (P<0.01). From d 0 to 21, ADG and FCR were not affected by feed quality but feeding pellets or increasing the duration of feeding the phase I diets improved FCR (P<0.01). Also, in this period PWD was higher with pellets than with mash and for pigs fed the phase I diets for 14 or 21 d than for pigs fed this diet for only 7 d (P<0.01). From d 21 to 35, pigs previously fed the LQ diet had higher ADG than pigs fed the HQ phase I diets (P<0.001). Also, pigs that were fed the phase I diets for 21 d had lower ADG (P<0.05) and ADFI (P< 0.001) and poor FCR (P<0.05) than pigs fed these diets for 7 or 14 d. Organic matter digestibility was higher for pigs fed the HQ phase I diets than for pigs fed the LQ phase I diets (P<0.05). Pelleting improved TTAD of all nutrients (P<0.01). It is concluded that HQ phase I diets increased TTAD of nutrients but not feed efficiency of Iberian pigs from d 28 to 63 d of age. Also, pelleting improved nutrient digestibility and feed efficiency. Increasing the duration of supply of the phase I diets from 7 to 21 d improved feed efficiency but reduced ADG. Therefore, the use of LQ phase I diets in pellet form for no more than 7 d after weaning is recommended in Iberian pigs.

Influence of source and micronization of soya bean meal on growth performance, nutrient digestibility and ileal mucosal morphology of Iberian piglets

The effects of inclusion in the diet of different sources of soya bean meal (SBM) on growth performance, total tract apparent digestibility (TTAD) and apparent ileal digestibility (AID) of major dietary components and mucosal ileum morphology were studied in Iberian pigs weaned at 30 days of age. From 30 to 51 days of age (phase I), there was a control diet based on regular soya bean meal (R-SBM; 44% CP) of Argentina (ARG) origin and five extra diets in which a high-protein soya bean meal (HP-SBM; 49% CP) of the USA or ARG origin, either ground (990 μm) or micronized (60 μm), or a soya protein concentrate (SPC; 65% CP) substituted the R-SBM. From 51 to 61 days of age (phase II), all pigs were fed a common commercial diet in mash form. The following pre-planned orthogonal contrasts were conducted: (1) R-SBM v. all the other diets, (2) SPC v. all the HP-SBM diets, (3) micronized HP-SBM v. ground HP-SBM, (4) HP-SBM of ARG origin v. HP-SBM of US origin and (5) interaction between source and the degree of grinding of the HP-SBM. Dietary treatment did not affect growth performance of the pigs at any age but from 30 to 51 days of age, post weaning diarrhoea (PWD) was higher (P<0.001) and the TTAD and AID of all nutrients were lower for pigs fed the R-SBM diet than for pigs fed the HP-SBM or the SPC diets. However, no differences between the HP-SBM and the SPC containing diets were detected for any trait. The TTAD of organic matter (P=0.07) and gross energy (GE) (P=0.05) tended to be higher for the micronized HP-SBM than for the ground HP-SBM and that of GE was higher (P<0.05) for US meal than for the ARG meal. Pigs fed R-SBM had lower villus height (P<0.01) than pigs fed HP-SBM or SPC but no differences in ileal mucosal morphology were detected between SPC and HP-SBM containing diets. It is concluded that feeding the HP-SBM or SPC-reduced PWD and improved nutrient digestibility and ileal morphology as compared with feeding the R-SBM, but had no effect on pig performance. The inclusion in the diet of added value soya products (micronized SBM or SPC) in substitution of the R-SBM increased the TTAD of all nutrients and reduced PWD but had no advantage in terms of growth performance over the use of ground HP-SBM.

Influence of source and micronization of soya bean meal on growth performance, nutrient digestibility and ileal mucosal morphology of Iberian piglets

The effects of inclusion in the diet of different sources of soya bean meal (SBM) on growth performance, total tract apparent digestibility (TTAD) and apparent ileal digestibility (AID) of major dietary components and mucosal ileum morphology were studied in Iberian pigs weaned at 30 days of age. From 30 to 51 days of age (phase I), there was a control diet based on regular soya bean meal (R-SBM; 44% CP) of Argentina (ARG) origin and five extra diets in which a high-protein soya bean meal (HP-SBM; 49% CP) of the USA or ARG origin, either ground (990 ?m) or micronized (60 ?m), or a soya protein concentrate (SPC; 65% CP) substituted the R-SBM. From 51 to 61 days of age (phase II), all pigs were fed a common commercial diet in mash form.

Vaccination with irradiated autologous melanoma cells engineered to secrete human granulocyte–macrophage colony-stimulating factor generates potent antitumor immunity in patients with metastatic melanoma

We conducted a Phase I clinical trial investigating the biologic activity of vaccination with irradiated autologous melanoma cells engineered to secrete human granulocyte–macrophage colony-stimulating factor in patients with metastatic melanoma. Immunization sites were intensely infiltrated with T lymphocytes, dendritic cells, macrophages, and eosinophils in all 21 evaluable patients. Although metastatic lesions resected before vaccination were minimally infiltrated with cells of the immune system in all patients, metastatic lesions resected after vaccination were densely infiltrated with T lymphocytes and plasma cells and showed extensive tumor destruction (at least 80%), fibrosis, and edema in 11 of 16 patients examined. Antimelanoma cytotoxic T cell and antibody responses were associated with tumor destruction. These results demonstrate that vaccination with irradiated autologous melanoma cells engineered to secrete granulocyte–macrophage colony-stimulating factor stimulates potent antitumor immunity in humans with metastatic melanoma.

Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation

Histone deacetylases (HDACs) catalyze the removal of acetyl groups on the amino-terminal lysine residues of core nucleosomal histones. This activity is associated generally with transcriptional repression. We have reported previously that inhibition of HDAC activity by hydroxamic acid-based hybrid polar compounds, such as suberoylanilide hydroxamic acid (SAHA), induces differentiation and/or apoptosis of transformed cells in vitro and inhibits tumor growth in vivo. SAHA is a potentially new therapeutic approach to cancer treatment and is in Phase I clinical trials. In several tumor cell lines examined, HDAC inhibitors alter the expression of less than 1% of expressed genes, including the cell cycle kinase inhibitor p21WAF1. In T24 bladder carcinoma cells, SAHA induces up to a 9-fold increase in p21WAF1 mRNA and protein, which is, at least in part, because of an increase in the rate of transcription of the gene. SAHA causes an accumulation of acetylated histones H3 and H4 in total cellular chromatin by 2 h, which is maintained through 24 h of culture. An increase in the accumulation of acetylated H3 and H4 was detected throughout the p21WAF1 promoter and the structural gene after culture with SAHA. The level of histone acetylation did not change in chromatin associated with the actin and p27 genes, and their mRNA expression was not altered during culture of T24 cells with SAHA. Thus, the present findings indicate that the induction of p21WAF1 by SAHA is regulated, at least in part, by the degree of acetylation of the gene-associated histones and that this induced increase in acetylation is gene selective.

Quorum-sensing acts at initiation of chromosomal replication in Escherichia coli

Chromosomal replication in Escherichia coli was studied by flow cytometry and was found to be inhibited by an extracellular factor present in conditioned media collected during late exponential and early stationary phase, i.e., via a quorum-sensing mechanism. Our results suggest that the inhibitory activity of the extracellular factor is exerted during initiation of DNA replication rather than during elongation. Furthermore, we present evidence that this interaction may occur directly at each of the replication forks. Unlike other quorum-sensing systems described so far for Gram-negative bacteria, this inhibitory activity does not require transcription or translation to be effective. Implications of quorum-sensing regulation of DNA replication are discussed.

p53-dependent cell cycle arrest induced by N-acetyl-L-leucinyl-L-leucinyl-L-norleucinal in platelet-derived growth factor-stimulated human fibroblasts.

Proteases are known to play important roles in cell growth control, although the underlying mechanisms are still poorly understood. Here we show that the protease inhibitor N-acetyl-L-leucinyl-L-leucinyl-L-norleucinal induced cell cycle arrest in platelet-derived growth factor-stimulated human fibroblasts at the G1/S boundary of the cell cycle by inhibiting the proteasome. Inhibition of the proteasome resulted in accumulation of the tumor suppressor p53, which was followed by an increase in the amount of the cyclin-dependent kinase-inhibitor p21. As a consequence, both phosphorylation and activity of the cyclin-dependent kinase 2/cyclin E complex were inhibited. We further observed that the retinoblastoma gene product, pRb, remained in the hypophosphorylated state, thus preventing cells from progression into the S-phase. These studies strongly support the hypothesis that the proteasome is a key regulator in the G1-phase of cell cycle progression.

Structural basis for specificity and potency of a flavonoid inhibitor of human CDK2, a cell cycle kinase.

The central role of cyclin-dependent kinases (CDKs) in cell cycle regulation makes them a promising target for studying inhibitory molecules that can modify the degree of cell proliferation. The discovery of specific inhibitors of CDKs such as polyhydroxylated flavones has opened the way to investigation and design of antimitotic compounds. A novel flavone, (-)-cis-5,7-dihydroxyphenyl-8-[4-(3-hydroxy-1-methyl)piperidinyl] -4H-1-benzopyran-4-one hydrochloride hemihydrate (L868276), is a potent inhibitor of CDKs. A chlorinated form, flavopiridol, is currently in phase I clinical trials as a drug against breast tumors. We determined the crystal structure of a complex between CDK2 and L868276 at 2.33 angstroms resolution and refined to an Rfactor 20.3%. The aromatic portion of the inhibitor binds to the adenine-binding pocket of CDK2, and the position of the phenyl group of the inhibitor enables the inhibitor to make contacts with the enzyme not observed in the ATP complex structure. The analysis of the position of this phenyl ring not only explains the great differences of kinase inhibition among the flavonoid inhibitors but also explains the specificity of L868276 to inhibit CDK2 and CDC2.

Disruption of cellular signaling pathways by daunomycin through destabilization of nonlamellar membrane structures.

Albeit anthracyclines are widely used in the treatment of solid tumors and leukemias, their mechanism of action has not been elucidated. The present study gives relevant information about the role of nonlamellar membrane structures in signaling pathways, which could explain how anthracyclines can exert their cytocidal action without entering the cell [Tritton, T. R. & Yee, G. (1982) Science 217, 248-250]. The anthracycline daunomycin reduced the formation of the nonlamellar hexagonal (HII) phase (i.e., the hexagonal phase propensity), stabilizing the bilayer structure of the plasma membrane by a direct interaction with membrane phospholipids. As a consequence, various cellular events involved in signal transduction, such as membrane fusion and membrane association of peripheral proteins [e.g., guanine nucleotide-binding regulatory proteins (G proteins and protein kinase C-alpha beta)], where nonlamellar structures (negative intrinsic monolayer curvature strain) are required, were altered by the presence of daunomycin. Functionally, daunomycin also impaired the expression of the high-affinity state of a G protein-coupled receptor (ternary complex for the alpha 2-adrenergic receptor) due to G-protein dissociation from the plasma membrane. In vivo, daunomycin also decreased the levels of membrane-associated G proteins and protein kinase C-alpha beta in the heart. The occurrence of such nonlamellar structures favors the association of these peripheral proteins with the plasma membrane and prevents daunomycin-induced dissociation. These results reveal an important role of the lipid component of the cell membrane in signal transduction and its alteration by anthracyclines.

Biomarcatori per il monitoraggio biologico di soggetti esposti ad agenti cancerogeni per il polmone

Riassunto I biomarcatori o “marcatori biologici” svolgono un ruolo fondamentale nel monitoraggio biologico. In questo lavoro ci siamo soffermati sullo studio di biomarcatori di effetto e di esposizione a xenobiotici ambientali. Nel primo caso abbiamo valutato i micro RNA (miRNA) da utilizzare per la diagnosi precoce del tumore al polmone in matrici di facile accesso, quale il CAE e il plasma, utilizzando il miRNA-21, oncogeno, e il miRNA-486, oncosoppressore. I risultati evidenziano una loro capacità di distinguere correttamente i soggetti con tumore polmonare dai soggetti sani, ipotizzando un loro utilizzo a scopo diagnostico. Nella seconda parte del lavoro di tesi sono stati studiati i biomarcatori di esposizione a benzene per valutare gli effetti dell’esposizione a concentrazioni ambientali su bambini residenti in città e a diverso livello di urbanizzazione. Lo studio ha evidenziato una correlazione dose-effetto fra le concentrazioni di benzene e dei suoi metaboliti urinari e un danno ossidativo a livello degli acidi nucleici. Tuttavia, le concentrazioni di benzene urinario non sono influenzate dal grado di industrializzazione, a differenza dell’S-PMA e degli indicatori di stress ossidativo (8-oxodGuo e 8-oxoGuo) che sembrano risentire sia della residenza che del momento del campionamento. Infine abbiamo ricercato possibili biomarcatori di esposizione a vinilcicloesene (VCH), sottoprodotto industriale nella polimerizzazione del 1,3-butadiene, poiché non sono ancora stati proposti BEI di riferimento nonostante i bassi valori di TLV-TWA (0.1 ppm) proposti dall’ACGIH. Nella prima fase del lavoro abbiamo studiato i meccanismi di tossicità del VCH tramite modelli in vitro, testando varie linee cellulari. I risultati evidenziano come la dose reale di VCH sia di molto inferiore a quella nominale per effetto dell’evaporazione. Inoltre, nelle linee cellulari più sensibili si sono evidenziati effetti citostatici, con alterazioni del ciclo cellulare, a differenza dell’esposizione agli epossidi del VCH, il VCD e l’1,2-VCHME, che determinano lisi cellulare con IC50 di 3 ordini di grandezza inferiori a quelli del VCH. La quantificazione dei metaboliti di I fase e di II fase del VCH nelle linee cellulari epatiche ha evidenziato concentrazioni di circa 1000 volte inferiori a quelle del VCH confermando come la sua tossicità sia principalmente dovuta alla produzione degli intermedi epossidici. La trasformazione nei metaboliti di II fase conferma inoltre l’effetto detossificante del metabolismo. La trasferibilità dei risultati ottenuti in vitro su sistemi in vivo fornirà le basi per poter identificare possibili metaboliti da proporre per il monitoraggio biologico di lavoratori esposti a VCH. PAROLE CHIAVE: biomarcatori di effetto e di esposizione, tumore al polmone, miRNA, benzene, vinilcicloesene.

Nitrato na dieta de ruminantes como estratégia nutricional para mitigação de metano entérico

A produção de metano entérico está entre as principais fontes de emissão de gases de efeito estufa dentre as atividades agropecuárias, além de gerar perda energética ao animal de até 12% da energia bruta consumida. Assim, o objetivo deste trabalho foi avaliar o uso de nitrato de cálcio encapsulado na alimentação de ruminantes como estratégia nutricional a mitigação de metano entérico. O experimento consistiu de duas fases. Fase I: Foram testadas dietas suplementadas com produto comercial de nitrato de cálcio encapsulado utilizando a técnica semiautomática de produção de gases in vitro. Meio grama de substrato com 50 mL de meio de incubação e 25 mL de inóculo ruminal foram incubados em frascos de vidro (160 mL) à 39 ºC por 24 horas para determinação da melhor dieta a ser testada in vivo. O primeiro ensaio testou a associação entre a monensina (dietas com e sem adição de monensina) e doses de nitrato encapsulado (0; 1,5 e 3% da matéria seca (MS)) para mitigação de metano in vitro. Não foi observada interação entre monensina e nitrato para as variáveis testadas. O segundo ensaio in vitro testou a interação do tipo de dieta com duas relações concentrado:volumoso, 20:80 e 80:20, e a inclusão de doses de nitrato encapsulado (0; 1,5; 3 e 4,5% MS). Embora não foi observado efeito associativo entre dieta e nitrato para redução de metano, foi observada mudança nos produtos da fermentação ruminal, com redução de propionato, em decorrência da concorrência de nitrato e propianogênicas por hidrogênio mais escasso em dietas com menor fermentação. Fase II: Conforme os resultados obtidos na Fase I, na segunda fase foi avaliado o efeito associativo da relação de concentrado:volumoso da dieta e a dose de nitrato sobre a emissão de metano, constituintes ruminais e toxicidade do nitrato in vivo. Utilizou-se seis borregos canulados no rúmen, distribuídos em delineamento experimental quadrado latino 6 x 6, em fatorial 2 x 3. Os fatores foram tipo de dieta (relação concentrado:volumoso 20:80 e 80:20) e inclusão de doses de nitrato encapsulado na dieta (0; 1,5 e 3% MS) em substituição gradual ao farelo de soja, totalizando seis tratamentos. Os teores de substituição do farelo de soja pelo nitrato foram em equivalente proteico de maneira a deixar as dietas isonitrogenadas. Os animais foram adaptados gradualmente a oferta de nitrato dietético para evitar problemas com toxidez. A análise de toxicidade foi avaliada pela taxa de metahemoglobina no sangue dos ovinos 3 horas após a alimentação. Nitrato reduziu a produção de metano em ambas as dietas. Os níveis de metahemoglobina no sangue dos animais não foram alterados pela adição de nitrato. Foi observado efeito associativo entre o tipo de dieta e nitrato para os produtos da fermentação ruminal, como acetato, que aumentou linearmente nas dietas com 80% de concentrado quando nitrato foi adicionado. Concluí-se que nitrato, utilizado de forma segura, é uma promissora estratégia para redução de metano entérico independentemente do tipo de dieta com que está sendo suplementado

Gender analysis of moxifloxacin clinical trials

Purpose: To determine the inclusion of women and the sex-stratification of results in moxifloxacin Clinical Trials (CTs), and to establish whether these CTs considered issues that specifically affect women, such as pregnancy and use of hormonal therapies. Previous publications about women’s inclusion in CTs have not specifically studied therapeutic drugs. Although this type of drug is taken by men and women at a similar rate, adverse effects occur more frequently in the latter. Methods: We reviewed 158 published moxifloxacin trials on humans, retrieved from MedLine and the Cochrane Library (1998–2010), to determine whether they complied with the gender recommendations published by U.S. Food and Drug Administration Guideline. Results: Of a total of 80,417 subjects included in the moxifloxacin CTs, only 33.7% were women in phase I, in contrast to phase II, where women accounted for 45%, phase III, where they represented 38.3% and phase IV, where 51.3% were women. About 40.9% (n = 52) of trials were stratified by sex and 15.3% (n = 13) and 9% (n = 7) provided data by sex on efficacy and adverse effects, respectively. We found little information about the influence of issues that specifically affect women. Only 3 of the 59 journals that published the moxifloxacin CTs stated that authors should stratify their results by sex. Conclusions: Women are under-represented in the published moxifloxacin trials, and this trend is more marked in phase I, as they comprise a higher proportion in the other phases. Data by sex on efficacy and adverse effects are scarce in moxifloxacin trials. These facts, together with the lack of data on women-specific issues, suggest that the therapeutic drug moxifloxacin is only a partially evidence-based medicine.