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.

Bile Acids Induce Glucagon-Like Peptide 2 Secretion with Limited Effects on Intestinal Adaptation in Early Weaned Pigs

Early weaning is a stressful event characterized by a transient period of intestinal atrophy that may be mediated by reduced secretion of glucagon-like peptide (GLP) 2. We tested whether enterally fed bile acids or plant sterols could increase nutrient-dependent GLP-2 secretion and improve intestinal adaptation in weanling pigs. During the first 6 d after weaning, piglets were intragastrically infused once daily with either deionized water -control-, chenodeoxycholic acid -CDC; 60mg/kg body weight-, or b-sitoesterol -BSE; 100 mg/kg body weight-. Infusing CDC increased plasma GLP-2 -P menor que 0.05- but did not affect plasma GLP-1 and feed intake. The intestinal expression of Glp2r -glucagon-like peptide 2 receptor-, Asbt -sodium-dependent bile acid transporter-, Fxr -farnesoid X receptor-, and Tgr5 -guanosine protein?coupled bile acid receptor- genes were not affected by CDC treatment. The intragastric administration of CDC did not alter the weight and length of the intestine, yet increased the activation of caspase-3 in ileal villi -P menor que 0.02- and the expression of Il6 -interleukin 6; P menor que 0.002- in the jejunum. In contrast, infusing BSE did not affect any of the variables that were measured. Our results show that the enteral administration of the bile acid CDC potentiates the nutrient-induced secretion of endogenous GLP-2 in early-weaned pigs. Bile acid?enhanced release of GLP-2, however, did not result in improved intestinal growth, morphology, or inflammation during the postweaning degenerative phase.

PV-1 is a component of the fenestral and stomatal diaphragms in fenestrated endothelia

PV-1 is a novel endothelial protein shown by immunocytochemical tests to be specifically associated with the stomatal diaphragms of caveolae in lung endothelium. Although the highest expression levels of both mRNA and protein are in the lung, PV-1 also has been found to be expressed in other organs. Using a specific antibody to the extracellular domain of PV-1, we have extended the survey on the presence of this protein at light and electron microscope level in several rat organs. Here we show that by immunofluorescence the antibody recognizes with high specificity the endothelium of the fenestrated peritubular capillaries of the kidney and those of the intestinal villi, pancreas, and adrenals. By immunolocalization at electron microscope level, the antibody recognizes specifically the diaphragms of the fenestrae and the stomatal diaphragms of caveolae and transendothelial channels in the endothelia of these vascular beds. No signal was detected in the continuous endothelium of the heart, skeletal muscle, intestinal muscularis, or brain capillaries or the nondiaphragmed fenestrated endothelium of kidney glomeruli. Taken together, our findings define the only antigen to be localized thus far in fenestral diaphragms. They also show that the stomatal diaphragms of caveolae and transendothelial channels and the fenestral diaphragms might be biochemically related, in addition to being morphologically similar structures.

Protection against hemorrhagic shock in mice genetically deficient in poly(ADP-ribose)polymerase

Hemorrhagic shock (HS) and resuscitation leads to widespread production of oxidant species. Activation of the enzyme poly(ADP-ribose) polymerase (PARP) has been shown to contribute to cell necrosis and organ failure in various disease conditions associated with oxidative stress. We tested the hypothesis whether PARP activation plays a role in the multiple organ dysfunction complicating HS and resuscitation in a murine model of HS and resuscitation by using mice genetically deficient in PARP (PARP−/−) and their wild-type littermates (PARP+/+). Animals were bled to a mean blood pressure of 45 mmHg (1 mmHg = 133 Pa) and resuscitated after 45 min with isotonic saline (2× volume of shed blood). There was a massive activation of PARP, detected by poly(ADP-ribose) immunohistochemistry, which localized to the areas of the most severe intestinal injury, i.e., the necrotic epithelial cells at the tip of the intestinal villi, and colocalized with tyrosine nitration, an index of peroxynitrite generation. Intestinal PARP activation resulted in gut hyperpermeability, which developed in PARP+/+ but not PARP−/− mice. PARP−/− mice were also protected from the rapid decrease in blood pressure after resuscitation and showed an increased survival time, as well as reduced lung neutrophil sequestration. The beneficial effects of PARP suppression were not related to a modulation of the NO pathway nor to a modulation of signaling through IL-6, which similarly increased in both PARP+/+ and PARP−/− mice exposed to HS. We propose that PARP activation and associated cell injury (necrosis) plays a crucial role in the intestinal injury, cardiovascular failure, and multiple organ damage associated with resuscitated HS.

DC-SIGNR, a DC-SIGN homologue expressed in endothelial cells, binds to human and simian immunodeficiency viruses and activates infection in trans

DC-SIGN, a C-type lectin expressed on the surface of dendritic cells (DCs), efficiently binds and transmits HIVs and simian immunodeficiency viruses to susceptible cells in trans. A DC-SIGN homologue, termed DC-SIGNR, has recently been described. Herein we show that DC-SIGNR, like DC-SIGN, can bind to multiple strains of HIV-1, HIV-2, and simian immunodeficiency virus and transmit these viruses to both T cell lines and human peripheral blood mononuclear cells. Binding of virus to DC-SIGNR was dependent on carbohydrate recognition. Immunostaining with a DC-SIGNR-specific antiserum showed that DC-SIGNR was expressed on sinusoidal endothelial cells in the liver and on endothelial cells in lymph node sinuses and placental villi. The presence of this efficient virus attachment factor on multiple endothelial cell types indicates that DC-SIGNR could play a role in the vertical transmission of primate lentiviruses, in the enabling of HIV to traverse the capillary endothelium in some organs, and in the presentation of virus to CD4-positive cells in multiple locations including lymph nodes.

Transgenic mice that overexpress the human trefoil peptide pS2 have an increased resistance to intestinal damage.

pS2 is a member of the trefoil peptide family, all of which are overexpressed at sites of gastrointestinal injury. We hypothesized that they are important in stimulating mucosal repair. To test this idea, we have produced a transgenic mice strain that expresses human pS2 (hpS2) specifically within the jejunum and examined the effect of this overexpression on proliferation and susceptibility to indomethacin-induced damage. A transgenic mouse was produced by microinjecting fertilized oocytes with a 1.7-kb construct consisting of rat intestinal fatty acid binding protein promoter (positions -1178 to +28) linked to full-length (490 bp) hpS2 cDNA. Screening for positive animals was by Southern blot analysis. Distribution of hpS2 expression was determined by using Northern and Western blot analyses and immunohistochemical staining. Proliferation of the intestinal mucosa was determined by assessing the crypt cell production rate. Differences in susceptibility to intestinal damage were analyzed in animals that had received indomethacin (85 mg/kg s.c.) 0-30 h previously. Expression of hpS2 was limited to the enterocytes of the villi within the jejunum. In the nondamaged intestine, villus height and crypt cell production rate were similar in transgenic and negative (control) litter mates. However, there was a marked difference in the amount of damage caused by indomethacin in control and transgenic animals in the jejunum (30% reduction in villus height in controls vs. 12% reduction in transgenic animals, P < 0.01) but the damage sustained in the non-hpS2-expressing ileal region was similar in control and transgenic animals. These studies support the hypothesis that trefoil peptides are important in stimulating gastrointestinal repair.

Mouse Col18a1 is expressed in a tissue-specific manner as three alternative variants and is localized in basement membrane zones.

We have isolated overlapping cDNAs encoding the N-terminal non-triple-helical region of mouse alpha 1(XVIII) collagen and shown that three different variants of alpha 1(XVIII) collagen exist. Each of the three variants shows characteristic tissue-specific expression patterns. Immunohistochemical studies show positive staining for alpha 1(XVIII) collagen along the basement membrane zones of vessels in the intestinal villi, the choroid plexus, skin, liver, and kidney. Thus, we conclude that alpha 1(XVIII) collagen may interact (directly or indirectly) with components in basement membrane zones or on the basal surface of endothelial/epithelial cells.

Loss of Apc heterozygosity and abnormal tissue building in nascent intestinal polyps in mice carrying a truncated Apc gene.

Mutations in the APC (adenomatous polyposis coli) gene appear to be responsible for not only familial adenomatous polyposis but also many sporadic cases of gastrointestinal cancers. Using homologous recombination in mouse embryonic stem cells, we constructed mice that contained a mutant gene encoding a product truncated at a 716 (Apc delta 716). Mendelian transmission of the gene caused most homozygous mice to die in utero before day 8 of gestation. The heterozygotes developed multiple polyps throughout the intestinal tract, mostly in the small intestine. The earliest polyps arose multifocally during the third week after birth, and new polyps continued to appear thereafter. Surprisingly, every nascent polyp consisted of a microadenoma covered with a layer of the normal villous epithelium. These microadenomas originated from single crypts by forming abnormal outpockets into the inner (lacteal) side of the neighboring villi. We carefully dissected such microadenomas from nascent polyps by peeling off the normal epithelium and determined their genotype by PCR: all microadenomas had already lost the wild-type Apc allele, whereas the mutant allele remained unchanged. These results indicate that loss of heterozygosity followed by formation of intravillous microadenomas is responsible for polyposis in Apc delta 716 intestinal mucosa. It is therefore unlikely that the truncated product interacts directly with the wild-type protein and causes the microadenomas by a dominant negative mechanism.

Efeito da dieta líquida sobre o desempenho de leitões neonatos e seus reflexos na matriz suína

Para ser competitivo atualmente, o sistema intensivo de produção de suínos deve estar pautado na eficiência. A fim de obter esta eficiência produtiva, o avanço genético das ultimas décadas buscou por fêmeas suínas cada vez mais prolíficas. A prolificidade contudo, veio acompanhada por uma queda no consumo voluntário de alimento por parte das fêmeas, bem como um aumento na produção de leite, e no número de leitões nascidos; o aumento da leitegada, levou a uma redução do peso ao nascimento e um aumento da heterogeneidade entre os leitões. Como forma de contornar o problema, são oferecidas aos leitões dietas formuladas com ingredientes de alto valor biológico a partir dos sete dias de vida, procurando suprir a demanda nutricional do animal durante o período de amamentação e preparar seu sistema digestório para o desmame. Contudo, grande parte das dietas formuladas para os leitões neonatos são oferecidas aos animais em sua forma sólida. Neste projeto, avaliamos os efeitos sobre a performance de leitões neonatos e da performance reprodutiva da fêmea suína do oferecimento de uma dieta líquida para os leitões neonatos, dieta esta que foi disponibilizada aos animais através de um sistema automatizado que realizou a mistura do alimento em sua forma sólida com a água. Para tais avaliações, os leitões ao nascer foram alocados em três grupos distintos, recebendo a dieta em sua forma líquida, em sua forma sólida ou então apenas o leite materno. Foram avaliadas variáveis zootécnicas relacionadas aos leitões, como peso, ganho diário de peso, consumo de ração, conversão alimentar, mortalidade pré-desmame; frequência de dias com diarreia nos leitões em fase de maternidade e creche. Foram eutanasiados leitões aos 14 e aos 28 dias de idade, para a realização do exame morfométrico da altura de vilosidade, profundidade de cripta e a relação entre a altura de vilosidade e profundidade de cripta nas porções do duodeno, jejuno e íleo. Avaliamos também o impacto do uso da dieta líquida sobre o catabolismo sofrido pela fêmea durante a lactação, através da aferição do peso e da espessura de toucinho desta fêmeas durante o período lactacional e também a duração do intervalo desmame estro e a duração do estro subsequente ao desmame. Não verificamos contudo um melhor desempenho zootécnico dos leitões nos períodos de maternidade e creche, tão pouco uma alteração favorável quanto a frequência de dias com diarreia nas duas fases em relação aos leitões que não consumiram nenhum tipo de suplementação. Quanto aos parâmetros morfométricos do intestino delgado, apenas aos 28 dias de idade os leitões que receberam a dieta líquida apresentaram maiores alturas de vilosidades no íleo em relação aos leitões que consumiram a dieta sólida e os animais do grupo controle apresentaram menores profundidades de cripta no mesmo seguimento e idade quando comparados aos demais animais. Contudo, estas alterações não foram o suficiente para garantir diferenças na relação altura de vilosidade:profundidade de cripta. E ainda, a suplementação independente de sua forma não reduziu o catabolismo sofrido pela fêmea suína durante a lactação

Low dietary protein during early pregnancy alters bovine placental development

To determine if low dietary protein concentration in the first two trimesters of pregnancy alters placental development, genetically similar heifers from closed herd were fed diets containing different levels of protein in the first and second trimesters of gestation. There were four animals per treatment group, the groups being: L/L = fed a diet containing 7% crude protein (CP) (low protein) in the first and second trimesters; H/H = fed a diet containing 14% (P thigh protein) in the first and second trimesters; L/H = fed low protein in the first trimester and high in the second trimester and vice versa for the H/L group. Low protein diets in the first trimester increased dry cotyledon weight at term. Trophectoderm volume density increased in the H/L and L/H group compared to the L/L and H/H groups. Blood vessel volume and volume density in foetal villi decreased in the H/L and L/H groups compared with the H/H and L/L groups. There was no effect of diet treatment on cotyledon number, diameter or wet weight and no effect on the volume density of connective tissue or fibroblasts in the foetal villi. These results show that a low dietary protein concentration in the first trimester of pregnancy followed by increased protein in the second trimester enhanced placental development. Further, trophectoderm volume was highly correlated with birth weight. Early protein restriction in the pregnant cow may enhance foetal growth in part by stimulating placental growth and function. (C) 1999 Published by Elsevier Science B.V. All rights reserved.

Preimplantation genetic diagnosis: Analysis of gene expression, nuclear DNA and cytoplasmic DNA

Preimplantation genetic diagnosis (PGD) following in vitro fertilization (IVF) offers couples at risk for transmitting genetic disorders the opportunity to identify affected embryos prior to replacement. In particular, embryo gender determination permits screening for X-linked diseases of unknown etiology. Analysis of embryos can be performed by polymerase chain reaction (PCR) amplification of material obtained by micromanipulation. This approach provides an alternative to the termination of an established pregnancy following chorionic villi sampling or amniocentesis. ^ Lately, the focus of preimplantation diagnosis and intervention has been shifting toward an attempt to correct cytoplasmic deficiencies. Accordingly, it is the aim of this investigation to develop methods to permit the examination of single cells or components thereof for clinical evaluation. In an attempt to lay the groundwork for precise therapeutic intervention for age related aneuploidy, transcripts encoding proteins believed to be involved in the proper segregation of chromosomes during human oocyte maturation were examined and quantified. Following fluorescent rapid cycle RT-PCR analysis it was determined that the concentration of cell cycle checkpoint gene transcripts decreases significantly as maternal age increases. Given the well established link between increasing maternal age and the incidence of aneuploidy, these results suggest that the degradation of these messages in aging oocytes may be involved with inappropriate chromosome separation during meiosis. ^ In order to investigate the cause of embryonic rescue observed following clinical cytoplasmic transfer procedures and with the objective of developing a diagnostic tool, mtDNA concentrations in polar bodies and subcellular components were evaluated. First, the typical concentration of mtDNA in human and mouse oocytes was determined by fluorescent rapid cycle PCR. Some disparity was noted between the copy numbers of individual cytoplasmic samples which may limit the use of the current methodology for the clinical assessment of the corresponding oocyte. ^

Se on tuomionpäivä edessä. Akseligrammien edellä ja edessä käyttö ja metaforisuus

Tarkastelen pro gradu -tutkielmassani akseligrammien edellä ja edessä käyttöä ja metaforisuutta. Perusmerkityksessään edellä ja edessä ilmaisevat spatiaalisia eli tilaa koskevia suhteita (Matti on viisi metriä Teppoa edellä), mutta niitä voidaan käyttää myös temporaalisten (Kevään edellä oli pitkä talvi) ja muiden abstraktien (Suomi on kehityksessä Bangladeshista edellä) suhteiden kuvaamiseen. Abstraktien suhteiden kuvauksessa ete-grammien käyttö on kuitenkin aina metaforista. Tutkimukseni päätehtävä on kuvailla monipuolisesti edellä- ja edessä-grammien käyttöä sekä vertailla grammeja sisältävien metaforisten ja spatiaalisten ilmausten semantiikkaa ja rakennetta. Pro gradu -tutkielmani on jatkoa kandidaatintutkielmalleni (Teeri 2013), jossa selvitin edellä- ja edessä-grammien semantiikkaa spatiaalisia suhteita kuvaavissa ilmauksissa, joten aikaisemman tutkimukseni laajentaminen grammien edessä ja edellä käyttöön myös ei-spatiaalisessa merkitysympäristössä seuraa luonnollista jatkumoa. Tutkimusaineistokseni olen valinnut joukon lifestyle-blogeja, sillä uskon blogitekstin edustavan kirjoitetun arkikielen todellista käyttöä. Tutkimukseni on kvalitatiivinen ja pyrkii vastaamaan muun muassa siihen, miten edellä- ja edessä-muodot varioivat semanttisesti, millaisia metaforisia ilmauksia kielenkäyttäjät niistä muodostavat ja millaiset spatiaaliset kokemukset motivoivat niiden käyttöä metaforisissa ilmauksissa. Tutkimukseni pohjaa kognitiiviseen kielitieteeseen, jonka mukaan kieli on joukko konventionaalistuneita rakenneskeemoja, joilla kielenkäyttäjät kuvaavat maailmaa. Grammirakenteista muodostettujen spatiaalisten metaforien analyysissä apunani on kognitiivinen metaforateoria, jonka mukaan koko käsitejärjestelmämme on perusluonteeltaan metaforinen. Etenkin ajan spatiaaliset metaforat ovat herättäneet viime aikoina suurta kiinnostusta kielitieteilijöiden parissa, ja oma tutkimuksenikin painottuu suuresti juuri ajan metaforiin ja temporaalisten suhteiden kielentämiseen perusmerkitykseltään spatiaalisilla ete-grammeilla. Tutkimuksessani nousee esiin muun muassa se, että aineistossani edellä-grammi esiintyy metaforisessa ilmauksessa edessä-grammia useammin: edessä-grammi kuvaa abstraktia suhdetta noin 25 %:ssa koko aineiston ilmauksista, kun taas edellä-grammilla vastaava luku on hieman alle 50 %. Ajan spatiaalisista metaforista aineistossani käytetään useimmin LIIKKUVAN EGON metaforaa (20 %). Sen sijaan JONOMETAFORAA käytetään melko vähän (3 %). Tutkimus osoittaa sen, että käsitystämme ajasta tilana ohjaavat tarkat säännöt, joita kaikki tutkimani edellä- ja edessä-grammeja sisältävät ajan spatiaaliset metaforat noudattavat. Kaikki tutkimuksessani analysoidut metaforiset ilmaukset eivät kuitenkaan ole ajan spatiaalisia metaforia, vaan edellä-grammilla voidaan lisäksi jäsentää tekstiä sekä kuvata priorisoinnin kohdetta ja kehityksellistä tai saavutuksellista paremmuutta. Edessä-grammi ilmaisee temporaalisten ja spatiaalisten suhteiden ohella metaforista esteellisyyttä sekä auktoriteetin tai muun ihmiskasvoiseksi koetun abstraktin asian edessä olemista. Tutkimukseni avaa ovia akseligrammien tutkimuksen maailmaan ja mahdollistaa siirtymisen ete-grammien suuntasijamuotojen käytön ja metaforisuuden tutkimiseen. Mielenkiintoista olisi myös perehtyä vertikaalisiin akseligrammeihin ja niiden käyttöön ajanilmauksissa.

YHTEISTYÖTÄ VAI YLIMÄÄRÄISTÄ TYÖTÄ? Opettajien kokemuksia kolmiportaisen tuen asiakirjojen käytöstä esi- ja alkuopetuksen nivelvaiheessa

Suomessa esi- ja perusopetuksessa otettiin 1.1.2011 käyttöön uusi oppimisen kolmiportainen tukimalli, joka korostaa kaikkien lasten oikeutta oppimiseen ja koulunkäyntiin aivan tavallisilla luokilla. Tuen malli muotoutui vuoden 2010 perusopetuslakimuutoksessa. Tukimallin jokaiseen eri vaiheeseen kuuluu kiinteästi tuen toteuttamiseen liittyviä asiakirjoja. Niihin kirjataan lapsen tukeen liittyvät asiat. Asiakirjoja ovat: pedagoginen arvio, oppimissuunnitelma, pedagoginen selvitys sekä henkilökohtainen oppimisen järjestämistä koskeva suunnitelma (HOJKS). Tämän tutkimuksen tarkoitus oli tutkia asiakirjojen käytettävyyttä esi- ja alkuopetuksen nivelvaiheessa. Tutkimuksessa selvitettiin, millaisia kokemuksia esi- ja alkuopettajilla on kolmiportaisen tuen asiakirjoista sekä niiden vaikutuksista heidän yhteistyöhönsä. Tutkimukseen osallistui viisi esiopettajaa ja viisi alkuopettajaa. Tutkimus suoritettiin teemahaastatteluna fenomenologista lähestymistapaa noudattaen. Analyysimenetelmänä käytettiin aineistolähtöistä sisällönanalyysiä. Tutkimustulosten mukaan esi- ja alkuopettajat kokivat, että kolmiportaisen tuen asiakirjoihin kirjattu tieto vaikuttaa positiivisesti lapsen saamaan tukeen. Asiakirjat ohjaavat tukea tarvitsevan lapsen oppimisen polkua inkluusioperiaatteen mukaisesti kohti omaa lähikoulua. Opettajat näkivät kuitenkin asiakirjoihin liittyviä epäkohtia. Asiakirjat olivat epäselviä ja eri kunnissa hyvin erilaisia. Esiopettajat kokivat niiden kirjaamisen työläänä ja osittain turhana, sillä heillä ei ollut tietoa, lukevatko alkuopettajat heidän esiopetuksesta lähettämiään asiakirjoja. Tuen eri asteiden ”rajapinnat” olivat opettajille epäselviä. Ei ollut selvää, milloin esimerkiksi oppimissuunnitelma oli aiheellista kirjoittaa. Alkuopettajien mielestä lasten pulmia oli vaikea ennakoida, koska eri esiopettajat kirjasivat samoja asioita eri tavalla. Asiakirjoihin liittyvät yhteistyö- ja siirtokäytänteet eivät myöskään olleet selviä. Tutkimus osoitti, että kolmiportaisen tuen asiakirjoja pitäisi esiopetuksessa selkeyttää ja yhdenmukaistaa. Esimiesten tulisi antaa yhtenevät ohjeet asiakirjojen kirjaamis- ja siirtokäytänteisiin: kuka tekee mitä ja millä ajalla. Erityisopettajien työpanosta asiakirjojen suhteen tulisi myös tarkentaa. Opettajien lisäkoulutukseen oli selkeästi tarvetta.

Designing gainsharing model variations with logistics service company

This thesis studies, in collaboration with a Finnish logistics service company, gainsharing and the development of a gainsharing models in a logistics outsourcing context. The purpose of the study is to create various gainsharing model variations for the use of a service provider and its customers in order to develop and enhance the customer’s processes and operations, create savings and improve the collaboration between the companies. The study concentrates on offering gainsharing model alternatives for companies operating in internal logistics outsourcing context. Additionally, the prerequisites for the gainsharing arrangement are introduced. In the beginning of the study an extensive literature review is conducted. There are three main themes explored which are the collaboration in an outsourcing context, key account management and gainsharing philosophy. The customer expectations and experiences are gathered by interviewing case company’s employees and its key customers. In order to design the gainsharing model prototypes, customers and other experts’ knowledge and experiences are utilized. The result of this thesis is five gainsharing model variations that are based on the empirical and theoretical data. In addition, the instructions related to each created model are given to the case company, but are not available in this paper

Suhde- ja verkostolähtöinen näkökulma urheilutapahtuman sponsorointiin Case Paavo Nurmi Games

Urheilutapahtumat tavoittavat miljoonia ihmisiä päivittäin ympäri maailmaa. Yritykset ovat jo pitkään olleet kiinnostuneita yhteistyöstä urheilutapahtumien kanssa. Sponsoroinnin rahavirta on jatkanut kasvua, vaikka muun mainonnan kasvu on ollut hidasta. Tapahtumasponsorointiin käytetyn rahan määrä on viime vuosina kasvanut noin viisi prosenttia vuositasolla. Tutkielman tarkoituksena oli tarkastella urheilutapahtuman verkostoa ja suhteita tapahtuman pääyhteistyökumppanien näkökulmasta. Tutkimuksen tavoite oli jaettu kahteen osa-ongelmaan: 1. Miten urheilutapahtuman pääyhteistyökumppanit kokevat oman suhteensa sponsoroitavaan kohteeseen 2. Miten urheilutapahtuman pääyhteistyökumppanit kokevat oman asemansa verkostossa Ensimmäisenä teemana tutkielmassa tarkasteltiin tapahtumasponsoroinnin tavoitteita ja kahden välisen sponsorointisuhteen toimintaa. Toisena tarkasteltiin urheilutapahtuman verkoston muodostumista, toimintaa ja suhteita. Näiden kahden teeman pohjalta rakennettiin teoreettinen viitekehys tukemaan empiirisen tutkimuksen toteuttamista. Tutkimus on luonteeltaan laadullinen tapaustutkimus, jossa aineisto kerättiin kymmeneltä Paavo Nurmi Games –urheilutapahtuman pääyhteistyökumppanitason yritykseltä. Tutkimuksessa haastateltujen yritysten edustajien haastattelun lisäksi he myös piirsivät oman hahmotelmansa PNG-verkostosta. Näiden verkostokuvien keskinäinen vertailu tuki haastatteluissa kerättyä ja litteroitua aineistoa. Empiirisen tutkimuksen tuloksissa aineistoa käsiteltiin teoreettiseen viitekehykseen perustuviin teemoihin tukeutuen. Teemojen avulla esiteltiin pääyhteistyökumppanien kokemuksia PNG-verkoston eri toimijoista, suhteista sekä tavoitteista sponsorointisuhteelle. Aineistosta havaittiin selkeitä yhtäläisyyksiä pääyhteistyökumppanien yhteistyön tavoitteissa sekä koetusta suhteesta sponsoroitavaan kohteeseen. Haastateltujen yritysten kokemukset omasta asemastaan verkostossa poikkesivat suuremmin toisistaan. Yhteistyön tavoitteet ja tausta vaikuttivat yrityksen kokemukseen verkostosta. Verkostosta tunnistettiin kuitenkin kaksi keskeisintä toimijaa erilaisista verkostokokemuksista huolimatta.