Enteral exposure to crude red kidney bean lectin induces maturation of the gut in suckling pigs.

The present investigation characterized the effect of red kidney bean lectin exposure on gut maturation and function in young piglets. Eleven suckling pigs were given by stomach tube a crude red kidney bean lectin preparation (containing about 25% lectin, 400 mg/kg BW) (lectin-treated pigs) at 10, 11, and 12 d of life, and an additional 16 pigs (control pigs) were given saline instead. On the next day, the intestinal absorptive capacity was determined in vivo, and on the 14th d of life the piglets were killed and organs and small intestine samples were collected for analyses and in vitro permeability experiments. The lectin-treated pigs showed an increase in stomach weights and mucosa thickness, whereas no weight effect was found for the small intestine, spleen, liver, or adrenals. Morphometric analyses of the small intestine in lectin-treated pigs showed a decrease in villus heights, an increase in crypt depths and crypt cell mitotic indices, and fewer vacuolated enterocytes per villus and reduced vacuole size. Lectin treatment also resulted in a decrease in the absorption of different-sized marker molecules after gavage feeding, a decrease in intestinal marker permeability, and a change in small intestinal disaccharidase activities, with increased maltase and sucrase activities. The size of the pancreatic acini was also greater in the lectin-treated pigs, but no increases in enzyme content or pancreatic weight could be determined. In addition, the blood plasma levels of cholecystokinin were higher in the lectin-treated than in the control pigs. The results indicate that exposure to crude red kidney bean lectin induces structural and functional maturation of the gut and pancreatic growth in young suckling piglets. This possibility of inducing gut maturation may lead to an improvement in the piglets' ability to adapt to weaning and to an increase in the growth and health of these animals.     

rDNA cytogenetics and some structural variability in an <Emphasis Type="Italic">Avena barbata</Emphasis> Pott ex Link × <Emphasis Type="Italic">A. sativa</Emphasis> subsp. <Emphasis Type="Italic">nuda</Emphasis> (L.) Gillet et Magne amphiploid after 5-azaC treatment

The paper reports on the study of DNA methylation patterns in chromosomes of an octoploid selection obtained from an amphiploid Avena barbata × Avena sativa ssp. nuda, their alteration and plant structural changes induced by 5-azacytidine (5-azaC) treatment. Subtelomere/telomere regions and some chromosomes remained heavily methylated after this practise. The increased concentration of 5-azaC strengthened the demethylation process. NOR rDNA loci on five and 5S rDNA doubled loci on three pairs of chromosomes were detected. Some 5S loci were colocalised with NORs. The number of active NORs was increased by one homologous pair after demethylation and this was related to minor loci. The NORs were mainly located on A/D genomes. Patterns of nucleoli fusion varied between main and lateral roots and demethylation increased the number of uninucleolar nuclei. Seed germination was inhibited and plant growth decreased under demethylation. 5-azaC also induced distinct anomalies in endosperm development. Extensive parts of the tissue were composed of defected cells or remained empty. Polyploidised clones of starchy or aleurone cells were observed, as were aleurone cells exhibiting new rare phenotypes.     

Antioxidative effect of quercetin and its equimolar mixtures with phenyltin compounds on liposome membranes

Our earlier studies have shown that the compounds diphenyltin dichloride (DPhT) and triphenyltin chloride (TPhT) in the presence of UVC radiation enhanced the degree of phosphatidylcholine liposome membrane oxidation (J. Agric. Food Chem. 2005, 53, 76-83). The prooxidative behavior of the compounds has now been confirmed with the electron paramagnetic resonance method, which proved the possibility that the studied compounds can exist in free radical forms. The present work investigates the possibility of the protective action of quercetin on phosphatidylcholine liposome membranes exposed to the prooxidative action of DPhT and TPhT induced by UV radiation (lambda = 253.7 nm). The concentrations of quercetin and its equimolar mixtures with DPhT and TPhT were determined (and compared with well-known antioxidants as standards-trolox and butylated hydroxytoluene, also in the presence of phenyltins) as those that induce 50% inhibition in oxidation of liposomes radiated with UV. They are 5.1 +/- 0.10, 2.9 +/- 0.12, and 1.9 +/- 0.08 microM (differences between the values are statistically significant), constituting the following sequence of antioxidative activity: quercetin:TPhT > quercetin:DPhT > quercetin. This relation is confirmed by the results on the antiradical ability of quercetin and its mixtures with DPhT and TPhT toward the free radical 1,1-diphenyl-2-pricrylhydrazil. Similar sequences obtained in both studies suggest a possible mechanism of the antiradical action of the mixtures as free radical scavengers. We suggested that (i) quercetin's ability, documented by spectrophotometric, infrared attenuated total reflectance spectroscopy, (1)H NMR, and molecular modeling methods, to form complexes with phenyltins indicates a possible way of protection against the peroxidation caused by the free radical forms of phenyltins and (ii) the differentiation in the action of the quercetin/TPhT and quercetin/DPhT associates (statisticaly significant) may result from a different localization in the liposome membrane, which is indicated by the results of the fluorimetric studies.     

Differences in monocarboxylic acid transporter type 1 expression in rumen epithelium of newborn calves due to age and milk or milk replacer feeding

The aim of the present study was to investigate whether besides age and solid feed intake, monocarboxylic acid transporter type 1 (MCT1) expression in the rumen epithelium of calves is affected by liquid feed type [whole milk (WM) or milk replacer (MR)]. Thirty bull calves at the mean age of 5 days were randomly allocated to five experimental groups (six calves/group). Six calves were slaughtered immediately after allocation to the trial (5 days of life), eighteen calves were fed MR and slaughtered at week intervals (on 12, 19, 26 days of life respectively), and six calves were fed WM and slaughtered at the 26 days of life. MCT1 protein abundance and the MCT1 mRNA level were investigated in the dorsal and ventral sack of the rumen. Solid feed intake and short‐chain fatty acids (SCFA) concentration in the rumen fluid increased linearly with calves' age. The amount of the MCT1 protein and mRNA in the dorsal sac of rumen as well as the amount of MCT1 protein in the cranial ventral sac of rumen also increased linearly with calves' age. Calves fed WM had greater solid feed intake in the last week of the study as compared to calves fed MR, but SCFA concentration in the rumen fluid was not different. MCT1 mRNA expression in the cranial dorsal sac of rumen and protein MCT1 expression in both dorsal and ventral cranial sack of the rumen were higher in calves fed WM as compared to calves fed MR. This study confirmed age‐dependent changes of MCT1 expression in the rumen epithelium of newborn calves and showed that its expression might be affected by liquid feed type.     

Histological and morphometric evaluation of the urethra and penis in male New Zealand White rabbits

This stereudy aimed at performing a histological and morphometric evaluation of the urethra and penis of male rabbits. Seven male New Zealand White rabbits weighing 2.1–3 kg were used in the study. The whole urethra, from the urinary bladder to the external urethral orifice, was dissected from the rabbits, and the tissue was sliced into sections at an interval of 2 mm. The sections were stained with haematoxylin–eosin, Masson–Goldner trichrome stain, Van Gieson's stain and Movat–Russell modified pentachrome stain. A detailed evaluation of the morphology and morphometry was performed. The parameters assessed were the type and height of epithelium, thickness and composition of connective tissue, and thickness and structure of muscularis. The histological structure of the rabbit urethra was found to be similar to humans. However, although the rabbits were found to have the same type of penis as the humans, the internal structure of the corpora cavernosa, the relative thickness of the tunica albuginea and the rudimentary glands of the penis were found to differ in these animals. The results of the present study may be useful in the designing of implants, drug testing or surgical procedures on the physiological and pathological urethra.     

High-pressure effects on the proteolytic enzymes of sea bass (Dicentrarchus labrax L.) fillets

High-pressure processing is a nonthermal technique ensuring food product safety and enabling a longer shelf life. The purpose of this experiment was to evaluate the effect of high pressure on the main proteolytic enzymes involved in fish muscle degradation during storage. Enzymes were extracted with sarcoplasmic proteins from Dicentrarchus labrax sea bass white muscle. Activity of cathepsins B, D, H, and L was quantified in protein extract, whereas calpain activity was evaluated after isolation from its endogenous inhibitor. High-pressure processing up to 500 MPa enhanced the activity of cathepsin B, H, and L, whereas the activity of cathepsin D increased up to 300 MPa and decreased above 300 MPa. With regard to calpain activity, high-pressure processing led to a decrease of activity, which was zero above 400 MPa. We suggest a leading explanation based on simultaneous deactivation of enzymes and an increase of liberation from lysosomes for cathepsins and on dissociation of subunits for calpains.     

A novel cytometric approach to study intestinal mucosa rebuilding in weaned pigs fed with dietary nucleotides

In the weaning period, transition from sow's milk to the post-weaning diet causes the withdrawal of important nutrients as milk nucleotides, which are known to be determinant for the development of the gastrointestinal tract and immune function. The role of dietary nucleotides in the rebuilding of gut epithelium remains unclear. The aim of the study was to evaluate using appropriate markers the rate of mitosis (Ki67) in the crypt epithelial cell, and the rate of apoptosis (active caspase 3), autophagy (MAP I LC3) and DNA damage (p53) in the crypt and villi epithelial cells of the small intestine of weaned pigs fed diets supplemented without/with nucleotides. An in-tissue cytometry method, based on confocal imaging and automated quantitative analysis was implemented. The studies allowed us to understand molecular bases of animal performance which could not be accessed by a routine histometric approach. Namely, the dietary nucleotides provided more uniform small intestine epithelium with considerably less animal-to-animal variation in respect to mitosis (SEM = 1.02 in control vs. 0.62 in supplemented group), autophagy (SEM = 1.22 vs. 0.99) and expression of p53 protein (SEM = 0.72 vs. 0.28). A significant increase in ratio of apoptosis and autophagy, and significantly lower p53 expression was found in the nucleotide-supplemented pigs as compared to control. The mitosis/apoptosis index was lower in the nucleotide-supplemented group suggesting slower rebuilding of gut epithelium in these pigs as compared to the control, respectively 0.454 to 0.191. Finally, in the supplemented group a drop in p53 expression was observed, however, it remains uncertain whether the reduction in DNA damage index is due to the supplementation with nucleotides utilized as a source for repair processes or due to the reduced DNA alterations.