RESUMEN
Gut apical amino acid (AA) transport activity is high at birth and during suckling, thus being essential to maintain luminal nutrient-dependent mucosal growth through providing AA as essential metabolic fuel, substrates and nutrient stimuli for cellular growth. Because system-B(0) Na(+)-neutral AA co-transporter (B(0)AT1, encoded by the SLC6A19 gene) plays a dominant role for apical uptake of large neutral AA including L-Gln, we hypothesized that high apical Na(+)-Gln co-transport activity, and B(0)AT1 (SLC6A19) in co-expression with angiotensin-converting enzyme 2 (ACE2) were expressed along the entire small intestinal crypt-villus axis in young animals via unique control mechanisms. Kinetics of Na(+)-Gln co-transport activity in the apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from liquid formula-fed young pigs, were measured with the membrane potential being clamped to zero using thiocyanate. Apical maximal Na(+)-Gln co-transport activity was much higher (p < 0.05) in the upper villus cells than in the middle villus (by 29 %) and the crypt (by 30 %) cells, whereas Na(+)-Gln co-transport affinity was lower (p < 0.05) in the upper villus cells than in the middle villus and the crypt cells. The B(0)AT1 (SLC6A19) mRNA abundance was lower (p < 0.05) in the crypt (by 40-47 %) than in the villus cells. There were no significant differences in B(0)AT1 and ACE2 protein abundances on the apical membrane among the upper villus, the middle villus and the crypt cells. Our study suggests that piglet fast growth is associated with very high intestinal apical Na(+)-neutral AA uptake activities via abundantly co-expressing B(0)AT1 and ACE2 proteins in the apical membrane and by transcribing the B(0)AT1 (SLC6A19) gene in the epithelia along the entire crypt-villus axis.
Asunto(s)
Sistemas de Transporte de Aminoácidos Básicos/biosíntesis , Sistemas de Transporte de Aminoácidos Neutros/biosíntesis , Alimentación Animal , Regulación de la Expresión Génica , Mucosa Intestinal/metabolismo , Yeyuno/metabolismo , Peptidil-Dipeptidasa A/biosíntesis , Enzima Convertidora de Angiotensina 2 , Animales , Femenino , Masculino , PorcinosRESUMEN
Global pig production contributes to about 35% of the world's meat production and consumption [...].
RESUMEN
Poor efficiency of dietary fibre utilization not only limits global pork production profit margin but also adversely affects utilization of various dietary nutrients. Poor efficiency of dietary nutrient utilization further leads to excessive excretion of swine manure nutrients and results in environmental impacts of emission of major greenhouse gases (GHG), odor, nitrate leaching and surface-water eutrophication. Emission of the major GHG from intensive pork production contributes to global warming and deteriorates heat stress to pigs in tropical and sub-tropical swine production. Exogenous fibre enzymes of various microbial cellulases, hemicellulases and pectinases have been well studied and used in swine production as the non-nutritive gut modifier feed enzyme additives in the past over two decades. These research efforts have aimed to improve growth performance, nutrient utilization, intestinal fermentation as well as gut physiology, microbiome and health via complementing the porcine gut symbiotic microbial fibrolytic activities towards dietary fibre degradation. The widely reported exogenous fibre enzymes include the singular use of respective cellulases, hemicellulases and pectinases as well as their multienzyme cocktails. The currently applied exogenous fibre enzymes are largely limited by their inconsistent in vivo efficacy likely due to their less defined enzyme stability and limited biochemical property. More recently characterized monomodular, multifunctional and processive endoglucanases have the potential to be more efficaciously used as the next-generation designer fibre biocatalysts. These newly emerging multifunctional and processive endoglucanases have the potential to unleash dietary fibre sugar constituents as metabolic fuels and prebiotics, to optimize gut microbiome, to maintain gut permeability and to enhance performance in pigs under a challenged environment as well as to parallelly unlock biomass to manufacture biofuels and biomaterials.
RESUMEN
Gut alkaline phosphatases (AP) dephosphorylate the lipid moiety of endotoxin and other pathogen-associated-molecular patterns members, thus maintaining gut eubiosis and preventing metabolic endotoxemia. Early weaned pigs experience gut dysbiosis, enteric diseases and growth retardation in association with decreased intestinal AP functionality. However, the role of glycosylation in modulation of the weaned porcine gut AP functionality is unclear. Herein three different research approaches were taken to investigate how deglycosylation affected weaned porcine gut AP activity kinetics. In the first approach, weaned porcine jejunal AP isoform (IAP) was fractionated by the fast protein-liquid chromatography and purified IAP fractions were kinetically characterized to be the higher-affinity and lower-capacity glycosylated mature IAP (p < 0.05) in comparison with the lower-affinity and higher-capacity non-glycosylated pre-mature IAP. The second approach enzyme activity kinetic analyses showed that N-deglycosylation of AP by the peptide N-glycosidase-F enzyme reduced (p < 0.05) the IAP maximal activity in the jejunum and ileum and decreased AP affinity (p < 0.05) in the large intestine. In the third approach, the porcine IAP isoform-X1 (IAPX1) gene was overexpressed in the prokaryotic ClearColiBL21 (DE3) cell and the recombinant porcine IAPX1 was associated with reduced (p < 0.05) enzyme affinity and maximal enzyme activity. Therefore, levels of glycosylation can modulate plasticity of weaned porcine gut AP functionality towards maintaining gut microbiome and the whole-body physiological status.
RESUMEN
Development of highly efficacious exogenous fibre degradation enzymes can enhance efficiency of dietary fibre utilization and sustainability of global pork production. The objectives of this study were to investigate in vitro stability for two processive endoglucanases, referred to as GH5-tCel5A1 and GH5-p4818Cel5_2A that were overexpressed in CLEARCOLIBL21(DE3). Three-dimensional models predicted presence of Cys residues on the catalytic site surfaces of GH5-tCel5A1 and GH5-p4818Cel5_2A; and time course experimental results shown that both cellulases were susceptible to auto-oxidation by airborne O2 and were unstable. Furthermore, we examined these endoglucanases' stability under the mimicked in vitro porcine gastric and the small intestinal pH and proteases' conditions. Eadie-Hofstee inhibition kinetic analyses showed that GH5-tCel5A1 and GH5-p4818Cel5_2A respectively lost 18 and 68% of their initial activities after 2-h incubations under the gastric conditions and then lost more than 90% of their initial activities after 2-3 h of incubations under the small intestinal conditions. Therefore, further enzyme protein engineering to improve resistance and alternatively post-fermentation enzyme processing such as coating to bypass the gastric-small intestinal environment will be required to enable these two processive endoglucanases as efficacious exogenous fibre enzymes in pig nutrition application.
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Celulasa , Celulasas , Animales , Dominio Catalítico , Celulasa/metabolismo , Celulasas/metabolismo , Celulosa/metabolismo , Fibras de la Dieta , PorcinosRESUMEN
Gut apical Na(+)-glucose cotransporter 1 (SGLT1) activity is high at the birth and during suckling, thus contributing substantially to neonatal glucose homeostasis. We hypothesize that neonates possess high SGLT1 maximal activity by expressing apical SGLT1 protein along the intestinal crypt-villus axis via unique control mechanisms. Kinetics of SGLT1 activity in apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from neonatal piglets by the distended intestinal sac method, were measured. High levels of maximal SGLT1 uptake activity were shown to exist along the jejunal crypt-villus axis in the piglets. Real-time RT-PCR analyses showed that SGLT1 mRNA abundance was lower (P < 0.05) by 30-35% in crypt cells than in villus cells. There were no significant differences in SGLT1 protein abundances on the jejunal apical membrane among upper villus, middle villus, and crypt cells, consistent with the immunohistochemical staining pattern. Higher abundances (P < 0.05) of total eukaryotic initiation factor 4E (eIF4E) protein and eIE4E-binding protein 1 γ-isoform in contrast to a lower (P < 0.05) abundance of phosphorylated (Pi) eukaryotic elongation factor 2 (eEF2) protein and the eEF2-Pi to total eEF2 abundance ratio suggest higher global protein translational efficiency in the crypt cells than in the upper villus cells. In conclusion, neonates have high intestinal apical SGLT1 uptake activity by abundantly expressing SGLT1 protein in the epithelia and on the apical membrane along the entire crypt-villus axis in association with enhanced protein translational control mechanisms in the crypt cells.
Asunto(s)
Animales Recién Nacidos/metabolismo , Mucosa Intestinal/metabolismo , Yeyuno/metabolismo , Transportador 1 de Sodio-Glucosa/metabolismo , Animales , Mucosa Intestinal/citología , Cinética , PorcinosRESUMEN
Expression of the small intestinal alkaline phosphatase (IAP) is enterocyte differentiation dependent and plays essential roles in the detoxification of pathogenic bacterial lipopolysaccharide endotoxin, maintenance of luminal pH, organic phosphate digestion, and fat absorption. This study was conducted to examine the effect of early weaning on adaptive changes in IAP digestive capacity (V(cap)) and IAP gene expression compared with suckling counterparts in pigs at ages 10-22 d. Weaning decreased (P < 0.05) IAP enzyme affinity by 26% and IAP maximal enzyme activity by 22%, primarily in the jejunal region, with the jejunum expressing 84-86% of the whole gut mucosal IAP V(cap) [mol/(kg body weight.d)]. The majority (98%) of the jejunal mucosal IAP maximal activity was associated with the apical membrane and the remaining (2%) existed as the intracellular soluble IAP. Weaning reduced the abundance of the 60-kDa IAP protein associated with the proximal jejunal apical membrane by 64% (P < 0.05). Furthermore, weaning reduced (P < 0.05) the relative abundance of the proximal jejunal IAP mRNA by 58% and this was in association with decreases (P < 0.05) in the abundances of cytoplasmic (27%) and nuclear (29%) origins of IAP caudal-associated homeobox transcription factor 1. In conclusion, early weaning decreased small intestinal IAP V(cap), IAP catalytic affinity, and IAP gene expression, and this may in part contribute to the susceptibility of early-weaned piglets to increased occurrence of enteric diseases and growth-check.
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Fosfatasa Alcalina/metabolismo , Regulación Enzimológica de la Expresión Génica/fisiología , Intestino Delgado/enzimología , Porcinos/metabolismo , Destete , Distribución por Edad , Fosfatasa Alcalina/genética , Crianza de Animales Domésticos , Animales , Femenino , MasculinoRESUMEN
It is well known that rapid gain of muscle mass in neonatal pigs is highly related to protein synthesis. However, the role of protein degradation in muscle gain of the neonatal period has not been well established. Calpains and their endogenous inhibitors, calpastatins, play a significant role in early-stage myofibrillar protein degradation. To investigate the role of calpain-calpastatin system in muscle protein accumulation, we studied the expressions of their mRNA in muscle tissue sampled at days 1, 4, 6, 12, 20 and 28 from a total of 36 neonatal pigs. The steady-state mRNA levels of calpains 1A, 2 and 3A, calpastatin types 1, 2 and 3, obtained by quantitative real-time PCR analysis, decreased by 2-4 folds at the age of 4 to 6 days compared to 1-day-old piglets. Then, the relatively low expression level was maintained through 28 days of age. Expressions of calpains 1A, 3A and calpastatin type 1 were significantly correlated with the measurements of muscle protein accumulations such as muscle protein content and RNA/protein ratio. Expressions of calpain 1A, calpastatin types 1 and 3 were negatively correlated with birth weight and fractional rate of growth. The levels of calpains 1A and 2 mRNA were correspondent to their protease activities. In conclusion, decreased levels of calpain and calpastatin expressions over development in neonatal pigs are associated with high protein accumulations, suggesting that dramatic muscle growth during the neonatal period may be partially controlled by down-regulated calpain-calpastatin system.
Asunto(s)
Proteínas de Unión al Calcio/genética , Calpaína/genética , Perfilación de la Expresión Génica , Músculo Esquelético/metabolismo , Animales , Animales Recién Nacidos , Proteínas de Unión al Calcio/metabolismo , Calpaína/metabolismo , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sus scrofaRESUMEN
Cellulases play important roles in the dietary fibre digestion in pigs, and have multiple industrial applications. The porcine intestinal microbiota display a unique feature in rapid cellulose digestion. Herein, we have expressed a cellulase gene, p4818Cel5_2A, which singly encoded a catalytic domain belonging to glycoside hydrolase family 5 subfamily 2, and was previously identified from a metagenomic expression library constructed from porcine gut microbiome after feeding grower pigs with a cellulose-supplemented diet. The activity of purified p4818Cel5_2A was maximal at pH 6.0 and 50 °C and displayed resistance to trypsin digestion. This enzyme exhibited activities towards a wide variety of plant polysaccharides, including cellulosic substrates of avicel and solka-Floc®, and the hemicelluloses of ß-(1 â 4)/(1 â 3)-glucans, xyloglucan, glucomannan and galactomannan. Viscosity, reducing sugar distribution and hydrolysis product analyses further revealed that this enzyme was a processive endo-ß-(1 â 4)-glucanase capable of hydrolyzing cellulose into cellobiose and cellotriose as the primary end products. These catalytic features of p4818Cel5_2A were further explored in the context of a three-dimensional homology model. Altogether, results of this study report a microbial processive endoglucanase identified from the porcine gut microbiome, and it may be tailored as an efficient biocatalyst candidate for potential industrial applications.
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Bacterias/aislamiento & purificación , Celulasa/metabolismo , Celulosa/metabolismo , Polisacáridos/metabolismo , Animales , Bacterias/enzimología , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Dominio Catalítico , Celobiosa/metabolismo , Celulasa/química , Celulasa/genética , Microbioma Gastrointestinal , Modelos Moleculares , Conformación Proteica , Especificidad por Sustrato , Porcinos , Triosas/metabolismoRESUMEN
There is dearth of knowledge with regards to mineral digestibility of ingredients in canines, and current knowledge is focused on the digestibility of supplemented minerals, not on intrinsic mineral digestibility of ingredients. The objectives of the present study were to determine the apparent and true digestibility (TD) of macronutrients and micronutrients, and the total tract gastrointestinal endogenous nutrient outputs in canines fed either animal- or vegetable-based adult maintenance diets. Eight purpose bred Beagles (two intact males, six spayed females) of similar age (2.12 ± 0.35 yr, mean ± SD) and weight (9.92 ± 0.73 kg, mean ± SD) were pair housed in kennels but fed individually based on individual maintenance energy requirements. Two basal diets (animal and vegetable protein based) were formulated to meet nutritional requirements of adult canines. Two additional trial diets were created, using the basal diets, by diluting diets by 50% with anhydrous α-d-glucose to attempt to quantify endogenous mineral losses and enable calculation of TD. All diets contained titanium dioxide at 0.3% for calculations of nutrient digestibility. Dogs were provided with deionized water as their only source of water throughout the trial. Dogs in a specific kennel were randomly assigned to an experimental diet for 10 d (experimental period), and fecal samples were collected the last 4 d of each period. All dogs were fed all experimental diets in random order based on a 4 × 4 replicated Latin square design. Dogs fed intact diets had a higher apparent mineral digestibility compared to dogs fed diluted diets (P < 0.05). Apparent phosphorus digestibility was higher for dogs fed the diet 2 compared with the diet 1 (P = 0.01) and the diluted diets (P < 0.001). There was a trend towards a greater TD of Cu for dogs fed the diet 2 compared with the diet 1 (P = 0.08). P, Mg, Zn, and Mn true digestibilities were higher for dogs fed the diet 2 compared with the diet 1 (P < 0.05, P = 0.01, P = 0.02, P = 0.009, respectively). These results suggest that apparent and TD do not result in similar values. Further research should be conducted on TD in canines only if a better model is developed.
Asunto(s)
Alimentación Animal/normas , Proteínas en la Dieta/clasificación , Digestión , Perros/fisiología , Minerales/metabolismo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Proteínas en la Dieta/análisis , Heces/química , Femenino , Tracto Gastrointestinal/fisiología , Masculino , Nutrientes , Fósforo Dietético , Distribución Aleatoria , VerdurasRESUMEN
Fractional protein synthesis rates (FSR) are widely measured by the flooding dose technique via either an i.g. or an i.v. route. This study was conducted to compare differences in tracer incorporation and FSR in organs and tissues of fed piglets. The piglets were surgically implanted with catheters and randomly assigned to receive a flooding dose of Phe (1.5 mmol/kg body weight, 40 percent molar enrichment with [(2)H(5)]Phe) in saline administered via an i.p. or an i.v. route. [(2)H(5)]Phe free-pool enrichment in plasma increased logarithmically (P < 0.05) from 0 to 25% in the i.p. group, whereas it rose to a peak level within 3 min of the tracer injection and then decreased linearly (P < 0.05) in the i.v. group. Intracellular free-pool tracer enrichments in organs and tissues were within the range of the values measured for the plasma-free pool (25-27%), reaching the flooding status. Administration of the tracer via the i.p. and i.v. routes induced a logarithmical pattern (P < 0.05) of a surge in plasma cortisol concentrations within 30 min. Measurements of FSR in plasma, cardiac muscle, and skeletal muscles were lower (P < 0.05) in the i.p. than in the i.v. group due to the adverse effect of cortisol surge being more dramatic (P < 0.05) in the i.p. than in the i.v. group at 30 min of the post-tracer administration. We conclude that FSR may be measured by the flooding dose through an i.p. or an i.v. route and the i.p. route may underestimate FSR by the flooding dose for plasma, cardiac muscle, and skeletal muscles. This concern may be addressed by a fast regimen of sampling to be completed within 12-20 min after an i.p. route of tracer injection.
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Fenilalanina/metabolismo , Proteínas/metabolismo , Animales , Peso Corporal , Deuterio , Femenino , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Marcaje Isotópico , Cinética , Especificidad de Órganos , Fenilalanina/administración & dosificación , PorcinosRESUMEN
Nonpoint phosphorus (P) pollution from animal manure is becoming a serious global problem. The current solution for the swine industry includes the enzyme phytase as a component in oil meal and cereal grain-based swine diets. A long-term approach is the production of transgenic phytase pigs that express phytase in the salivary glands and secrete it in the saliva. This study provides a detailed comparison of chemical structures of manure from conventional pigs and transgenic pigs that express phytase under growing and finishing phases using new solid-state NMR techniques. Spectral editing techniques and quantitative NMR techniques were used to identify and quantify specific functional groups. Two-dimensional (1)H- (13)C heteronuclear correlation NMR was used to detect their connectivity. Manure from conventional and transgenic pigs had similar peptide, carbohydrate, and fatty acid components, while those from transgenic pigs contained more carbohydrates and fewer nonpolar alkyls. There was no consistent effect from diets with or without supplemental phosphate or growth stages.
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6-Fitasa/genética , Animales Modificados Genéticamente/metabolismo , Estiércol/análisis , Porcinos/metabolismo , 6-Fitasa/metabolismo , Animales , Animales Modificados Genéticamente/genética , Carbohidratos/análisis , Ácidos Grasos/análisis , Expresión Génica , Espectroscopía de Resonancia Magnética , Péptidos/análisis , Glándulas Salivales/enzimología , Porcinos/genética , Porcinos/crecimiento & desarrolloRESUMEN
The hypocholesterolemic effects associated with soluble fiber consumption are clear from animal model and human clinical investigations. Moreover, the modulation of whole-body cholesterol metabolism in response to dietary fiber consumption, including intestinal cholesterol absorption and fecal sterol and bile acid loss, has been the subject of many published reports. However, our understanding of how dietary fibers regulate molecular events at the gene/protein level and alter cellular cholesterol metabolism is limited. The modern emphasis on molecular nutrition and rapid progress in 'high-dimensional' biological techniques will permit further explorations of the role of genetic polymorphisms in determining the variable interindividual responses to soluble fibers. Furthermore, with traditional molecular biology tools and the application of 'omic' technology, specific insight into how fibers modulate the expression of genes and proteins that regulate intestinal cholesterol absorption and alter hepatic sterol balance will be gained. Detailed knowledge of the molecular mechanisms by which soluble fibers reduce plasma cholesterol concentrations is paramount to developing novel fiber-based "cocktails" that target specific metabolic pathways to gain maximal cholesterol reductions.
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Colesterol/metabolismo , Fibras de la Dieta/uso terapéutico , Suplementos Dietéticos , Galactanos/uso terapéutico , Hipercolesterolemia/terapia , Mananos/uso terapéutico , Gomas de Plantas/uso terapéutico , Animales , Investigación Biomédica/métodos , Fibras de la Dieta/historia , Suplementos Dietéticos/historia , Galactanos/historia , Genómica , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Hipercolesterolemia/metabolismo , Hipercolesterolemia/fisiopatología , Absorción Intestinal , Hígado/metabolismo , Mananos/historia , Fenómenos Fisiológicos de la Nutrición , Gomas de Plantas/historia , Proteómica , Resultado del TratamientoRESUMEN
Previously, we have found phosphopeptides (PPPs) from hen egg yolk phosvitin possess a potent antioxidative activity against oxidative stress in human intestinal epithelial cells, Caco-2. However, their biological activity at the cellular level has not yet fully understood. The objective of this study is to evaluate the regulation of glutathione (GSH) biosynthesis-associated and antioxidant enzymes against oxidative stress in Caco-2 cells using an in vitro model. Treatment of 1 mM H2O2-induced Caco-2 cells with PPPs increased cellular GSH levels, concomitant with a significant increase in gamma-glutamylcysteine synthetase (gamma-GCS) activity and the expression of gamma-GCS heavy subunit mRNA. Furthermore, intracellular glutathione reductase, glutathione S-transferase, and catalase activities were elevated by PPPs. In addition, PPPs with high content of phosphorus showed higher induction of these enzyme activities than PPPs without phosphorus. These data indicate that oligophosphopeptides from hen egg yolk phosvitin can up-regulate cellular GSH biosynthesis-associated enzymes activity and antioxidative activities, which play key roles against tissue oxidative stress in the human intestinal epithelial cells.
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Antioxidantes/análisis , Yema de Huevo/química , Glutamato-Cisteína Ligasa/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fosfopéptidos/farmacología , Fosvitina/química , Células CACO-2 , Glutamato-Cisteína Ligasa/genética , Humanos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Composting is an environmentally sound method for the disposal of on-farm livestock mortalities that generates material suitable for use as fertilizer; however, this method is not generally permitted for disposal of transgenic livestock mortalities during the research and development phase. This study has explored the application of the polymerase chain reaction (PCR) as a method for assessing the persistence of transgene and mitochondrial DNA markers during the composting of euthanized transgenic pig. There was at least a 10(7) fold reduction of genetic material to a level that not either transgene or mitochondrion markers were detectable. At the end of the composting period, only bone fragments that were completely demineralised and chalky were detected. Chemically the compost was similar to that from pig litter and poultry mortalities, except the copper content was lower. Based on these data, composting appears to be an appropriate method for the disposal of transgenic animals.
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ADN Mitocondrial/genética , Suelo/análisis , Transgenes , 6-Fitasa/genética , Animales , Animales Modificados Genéticamente , ADN/aislamiento & purificación , Marcadores Genéticos , Sustancias Húmicas/análisis , Diente Molar/química , Reacción en Cadena de la Polimerasa , Porcinos/genéticaRESUMEN
The protective effects of hen egg yolk phosvitin phosphopeptides (PPPs) against hydrogen peroxide (H2O2)-induced oxidative stress were evaluated in an in vitro assay using human intestinal epithelial cells. Caco-2 cells were stimulated with 1 mM H2O2 for 6 h, and the secretion of IL-8, a proinflammatory mediator, was determined by ELISA as a biomarker of oxidative stress. The inhibition of H2O2-induced IL-8 secretion from Caco-2 cells was observed by pretreatment for 2 h with PPPs, but not with phosvitin. PPPs also suppressed the formation of malondialdehyde in H2O2-treated Caco-2 cells. Furthermore, intracellular glutathione levels and glutathione reductase activity were elevated by the addition of PPPs. The protective effects of PPPs against H2O2-induced oxidative stress were almost the same as that of glutathione, and PPPs with a high content of phosphorus exhibited higher protective activity than PPPs without phosphorus; however, phosphoserine itself did not show any significant antioxidative stress activity. These findings suggest that oligophosphopeptides from hen egg yolk phosvitin possess novel antioxidative activity against oxidative stress in intestinal epithelial cells and that phosphorus and peptide structure seem to have a key role in the activity.
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Yema de Huevo/química , Estrés Oxidativo/efectos de los fármacos , Fosfopéptidos/farmacología , Fosvitina/química , Animales , Células CACO-2 , Pollos , Femenino , Glutatión/análisis , Glutatión Reductasa/análisis , Humanos , Peróxido de Hidrógeno/farmacología , Interleucina-8/metabolismo , Malondialdehído/metabolismo , Fosfoserina/farmacología , Fosvitina/farmacologíaRESUMEN
This study was to examine the time course of sample-specific linearity of intrinsic phytase hydrolysis in major cereal grains and in ileal digesta and fecal samples and to determine the time course of the microbial phytase-catalyzed hydrolysis of various sources of phytate for estimating phytate phosphorus (P) content. The intrinsic phytase activity in barley, corn, oat, and wheat samples was measured over multiple time points from 0 to 120 min at 1.5 mmol.L(-1) of sodium phytate at pH 5.5 and 37 degrees C. Time courses of hydrolysis of purified phytate and phytate associated with the cereal grain samples and the pig digesta and fecal samples were examined with the Natuphos microbial phytase over multiple time points from 0 to 48 h of incubation. The intrinsic phytase hydrolysis was linear (P < 0.05) for up to 120 min for the barley, corn, and wheat samples, whereas in the oat sample the hydrolysis was linear (P < 0.05) for only up to 30 min of incubation. The intrinsic phytase activities (phytase unit: mumol.kg(-1) of dry matter.min(-1)) for the barley, corn, and wheat samples were estimated to be 693, 86, and 1189 by linear regression analysis. Intrinsic phytase activity (412 phytase units) for the oat sample based on a 30-min incubation was considerably higher than the value (103 phytase units) determined from the 120-min incubation for the same oat sample. There were quadratic with plateau relationships (P < 0.05) between the hydrolytic release of inorganic P from various sources of phytate and the incubation time. The minimal incubation times required for the complete hydrolysis of phytate were estimated to be 4, 3, and 11 h for the purified phytate, the cereal grain samples, and the pig digesta and feces, respectively. It was concluded that multiple time point experiments need to be conducted to determine valid intrinsic phytase activity and phytate P content in samples through intrinsic and microbial phytase hydrolysis incubations.
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6-Fitasa/metabolismo , Digestión , Grano Comestible/enzimología , Heces/química , Fósforo/análisis , Porcinos , 6-Fitasa/análisis , 6-Fitasa/química , Animales , Íleon/metabolismo , CinéticaRESUMEN
The study was to determine effects of dietary supplementation of chitosan (COS) and galacto-mannan-oligosaccharides (GMOS) on some serum biochemical indices, serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) levels, and hepatic and long gissimus muscle IGF-I mRNA expression in early-weaned piglets. Twenty six Duroc x Landrace x Yorkshire piglets at the age of 15 days were used. The piglets had access to creep feed during the suckling. Six piglets were sacrificed for sampling at the beginning of the study. The other 20 piglets were individually housed in metabolic cages and randomly allotted to four corn and soybean meal-based diets including the control group, the antibiotic group with 110 mg lincomycin/kg diet, the COS group containing 0.025% COS, and the GMOS group with 0.20% GMOS, respectively, in a 2-week feeding experiment. Blood urea nitrogen (BUN) level was reduced whereas serum total protein concentration was increased (P<0.05) in responses to the COS and GMOS supplementation. Dietary supplementation of COS and GMOS also increased (P<0.05) the serum GH and IGF-I levels along with enhanced hepatic and the muscle IGF-I mRNA abundance. Dietary supplementation of oligosaccharides such as COS and GMOS may improve growth and feed conversion efficiency by increasing plasma GH and IGF-I levels, in the early-weaned piglets.
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Quitosano/administración & dosificación , Factor I del Crecimiento Similar a la Insulina/biosíntesis , Mananos/administración & dosificación , Oligosacáridos/administración & dosificación , Porcinos/metabolismo , Animales , Animales Lactantes , Antibacterianos/administración & dosificación , Proteínas Sanguíneas/metabolismo , Nitrógeno de la Urea Sanguínea , Colesterol/sangre , Suplementos Dietéticos , Hormona del Crecimiento/sangre , Factor I del Crecimiento Similar a la Insulina/genética , Lincomicina/administración & dosificación , Hígado/efectos de los fármacos , Hígado/metabolismo , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Distribución Aleatoria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Porcinos/sangre , Triglicéridos/sangreRESUMEN
Use of feed antibiotics as growth promoters for control of pathogens associated with monogastric food animal morbidity and mortality has contributed to the development of antimicrobial resistance, which has now become a threat to public health on a global scale. Presently, a number of alternative feed additives have been developed and are divided into two major categories, including 1) the ones that are supposed to directly and indirectly control pathogenic bacterial proliferation; and 2) the other ones that are intended to up-regulate host gut mucosal trophic growth, whole body growth performance and active immunity. A thorough review of literature reports reveal that efficacy responses of current alternative feed additives in replacing feed antibiotics to improve performances and gut health are generally inconsistent dependent upon experimental conditions. Current alternative feed additives typically have no direct detoxification effects on endotoxin lipopolysaccharides (LPS) and this is likely the major reason that their effects are limited. It is now understood that pathogenic bacteria mediate their negative effects largely through LPS interactions with toll-like receptor 4, causing immune responses and infectious diseases. Therefore, disruptive biological strategies and a novel and new generation of feed additives need to be developed to replace feed antibiotic growth promoters and to directly and effectively detoxify the endotoxin LPS and improve gut health and performance in monogastric food animals.
RESUMEN
Long-term oxidative stress in the gastrointestinal tract can lead to the development of chronic intestinal disorders. Many food-derived antioxidants are effective in vitro, but the variable reports of in vivo efficacy and the pro-oxidant nature of some antioxidants necessitate alternative strategies for the reduction of in vivo oxidative stress. Compounds that up-regulate the production of endogenous antioxidants such as glutathione (GSH) and antioxidant enzymes provide novel approaches for the restoration of redox homeostatis. Egg yolk peptides (EYP) prepared from Alcalase and protease N digestion of delipidated egg yolk proteins were found to exhibit antioxidative stress properties. The effect of EYP supplementation was examined in a hydrogen peroxide-induced human colon cell line and in an animal model of intestinal oxidative stress. EYP significantly reduced the pro-inflammatory cytokine, IL-8, in Caco-2 cells. In piglets given intraperitoneal infusions of hydrogen peroxide, EYP treatment increased GSH and gamma-glutamylcysteine synthetase mRNA expression and activity, significantly increased antioxidant enzyme activities, in particular catalase and glutathione S-transferase activities, and reduced protein and lipid oxidation in the duodenum, jejunum, ileum, and colon. Furthermore, EYP boosted the systemic antioxidant status in blood by increasing the GSH concentration in red blood cells. These results suggest that EYP supplementation is a novel strategy for the reduction of intestinal oxidative stress.