Identification of marker genes for intestinal immunomodulating effect of a fructooligosaccharide by DNA microarray analysis

Prebiotic fructooligosaccharides are noted for their intestinal immunodulating effects, and the identification of markers for the effects is a matter of great concern. This study aimed to identify marker genes for physiological effects of a particular fructooligosaccharide (FOS) on a host animal and also to define the target of its function in the small intestine. DNA microarray technology was used to screen candidate marker genes, and comprehensive changes in gene expressions in the ileum of mice fed with FOS were investigated. One of the major physiological effects of FOS was intestinal immunomodulation. Marker genes were then identified for major histocompatibility complex classes I and II, interferon, and phosphatidylinositol metabolites. Also, the ileum was segmented into Peyer's patch (PP) and the other ileal organ (DeltaPP), and these were analyzed by quantitative RT-PCR method, with the result that the site for recognizing the FOS function was the DeltaPP rather than the PP. This is the first paper showing the markers for the physiological effects of FOS in the small intestine at gene expression level. Applying these marker genes would make it possible to clarify the mechanisms of how the administration of dietary FOS and associated changes in the intestinal environment are recognized by host organisms as well as how its immunomodulating effects are expressed in the body.     

Characterization of Murine Pituitary-Derived Cell Lines Tpit/F1, Tpit/E and TtT/GF

The pituitary is an important endocrine tissue of the vertebrate that produces and secretes many hormones. Accumulating data suggest that several types of cells compose the pituitary, and there is growing interest in elucidating the origin of these cell types and their roles in pituitary organogenesis. Therein, the histogenous cell line is an extremely valuable experimental tool for investigating the function of derived tissue. In this study, we compared gene expression profiles by microarray analysis and real-time PCR for murine pituitary tumor-derived non-hormone-producing cell lines TtT/GF, Tpit/F1 and Tpit/E. Several genes are characteristically expressed in each cell line: Abcg2, Nestin, Prrx1, Prrx2, CD34, Eng, Cspg4 (Ng2), S100β and nNos in TtT/GF; Cxcl12, Raldh1, Msx1 and Twist1 in Tpit/F1; and Cxadr, Sox9, Cdh1, EpCAM and Krt8 in Tpit/E. Ultimately, we came to the following conclusions: TtT/GF cells show the most differentiated state, and may have some properties of the pituitary vascular endothelial cell and/or pericyte. Tpit/F1 cells show the epithelial and mesenchymal phenotypes with stemness still in a transiting state. Tpit/E cells have a phenotype of epithelial cells and are the most immature cells in the progression of differentiation or in the initial endothelial-mesenchymal transition (EMT). Thus, these three cell lines must be useful model cell lines for investigating pituitary stem/progenitor cells as well as organogenesis.     

EFFECT OF IRON DEFICIENCY ON ROOT FERRIC CHELATE REDUCTASE,PROTON EXTRUSION,BIOMASS PRODUCTION AND MINERAL ABSORPTION OF CITRUS ROOT STOCK ORANGE JASMINE (MURRAYA EXOTICA L.)

Three-week iron (Fe) deficiency stress experiments were conducted using two citrus root stocks, Fe-deficiency tolerant Orange Jasmine (OJ, Murraya exotica L.) and the sensitive Flying Dragon [FD, Poncirus trifoliata var. monstrosa (T. Ito) Swingle]. Root ferric chelate reductase activity and proton extrusion increased in OJ between 12 and 18 d of stress, whereas there was no change in FD. Dry weight of OJ roots increased in contrast to FD which decreased. The Mn content in OJ remained the same even under Fe stress. Zn content in OJ roots doubled while that of FD increased 4-folds. The shoot/root Fe accumulation ratio increased in OJ while it decreased in FD. OJ apparently has mechanisms for increasing root biomass, controlling Fe reutilization and regulating manganese (Mn) and zinc (Zn) absorption in response to Fe deficiency. These mechanisms could help maintain homeostasis under heavy metal stress, which would be useful for improved growth of economically important citrus species.     

Effects of age, egg-laying activity, and Salmonella-inoculation on the expressions of gallinacin mRNA in the vagina of the hen oviduct

Gallinacins (Gal) are antimicrobial peptides that play significant roles in innate immunity in chickens. The aim of this study was to examine whether age of birds and egg-laying activity (laying and non-laying caused by feed-regulation) affect the mRNA expression of Gal-1, -2, and -3 in the vagina of hens, and whether their expressions are changed in response to the stimulation with Salmonella enteritidis (SE) and lipopolysaccharide (LPS). White Leghorn hens were divided into groups of young and old laying hens, and groups of laying and non-laying hens after feed-regulation. Vaginal cells were cultured and stimulated with SE or LPS. Expressions of Gal-1, -2, and -3 mRNA in their vaginal mucosa and cultured cells were examined by quantitative real-time RT-PCR. The expressions of Gal-1, -2, and -3 of the vaginal mucosa were significantly greater in old birds than in young birds. Expression of these Gals in the vagina were decreased in the regressed oviduct of non-laying birds compared with laying birds. The expressions of Gal-1, -2, and -3 in the cultured vaginal cells were increased by stimulation with SE or LPS within 24 h. These results suggest that the mRNA expressions of Gal-1, -2 and -3 in the vagina of laying hens increased with age, whereas they decreased in the regressed oviduct during the non-laying phase. Also, synthesis of these antimicrobial peptides in the vagina may increase in response to SE and LPS to eliminate SE bacteria.     

Characterization of monoclonal antibodies directed against the canine distemper virus nucleocapsid protein

We have established four monoclonal antibodies (MAbs) against the nucleocapsid protein (NP) of canine distemper virus (CDV). A competitive binding assay has revealed that the MAbs are directed against two antigenic domains. An immunofluorescence assay using a series of deletion clones of the NP and an immunoprecipitation assay using the NP have revealed that two of the MAbs recognize the C-terminal region of the NP while the other two recognize the tertiary structure of the N-terminal domain. These MAbs reacted with all eight strains of CDV used in this study, but showed different reactivities against measles virus and rinderpest virus.     

Effects of supplementary corn silage on the feed intake and milk production of time-restricted grazing dairy cows

The effects of supplementary corn silage (CS) of either 2 or 4 kg of dry matter (DM; S + 2 and S + 4, respectively) above the energy requirement for milk production and maintenance for grazing dairy cows (S) were determined. Time‐restricted grazing was used to compare the feed intake, milk production, and nitrogen and energy use of lactating cows. The experiment was carried out on two different pastures using a 3 × 3 Latin square design for each pasture. Cows were grazed for 5 h on a rotational grazing system and were fed concentrate (1 kg per 5 kg of milk yield). Herbage intake was measured using a weighing technique. To calculate the energy and nitrogen use, whole feces and urine were collected. There was no statistical effect of the pastures. Herbage intake decreased by the addition of CS (P = 0.02). The reduction of herbage DM intake per unit consumption of supplementary CS towards the S group were 0.80 and 0.45 kg for the S + 2 and S + 4 groups, respectively. The total DM intake for the S + 4 group was higher than that for the S and S + 2 groups (P = 0.02). Milk yield did not differ among treatments, even though the total DM intake for the S + 4 group was higher than that of the S and S + 2 groups. Nitrogen and energy use did not differ with the addition of CS.     

Middle upper beak fracture in a Red-crowned crane that completely recovered with external skeletal fixation

An adult Red-crowned crane in captivity that had a displaced compound fracture of the middle upper beak caused by an accident was treated by using Type I-a external skeletal fixator (ESF). The ESF that was equipped with a unilateral epoxy putty fixator and with five half-pins was fixed on the premaxilla bone. The crane with the ESF on the beak was able to feed smoothly by itself. The ESF was removed 49 days after the first surgery. Beak malocclusion, which was observed in the latter half period of fixing, spontaneously improved within one month after removal of the ESF. This is the first successful case of repair of an upper beak fracture in a Red-crowned crane.     

Effect of feeding non-fibrous carbohydrate before grazing on intake and nitrogen utilization in dairy cows throughout the grazing season

Ten lactating cows were used to determine the effect of feeding non-fibrous carbohydrate (NFC) supplement before grazing on feed intake and nitrogen (N) utilization throughout a grazing season. The experiment was conducted from June to September. Cows grazed twice a day (2.5 h × 2) under a set stocking system and were fed NFC supplement (1 kg/4 kg of milk yield) 2 h before grazing (PRE) or immediately after grazing (POST). Cows were also fed a grass and corn silage mixture ad libitum. Herbage dry matter intake (DMI) was greater for PRE than for POST throughout the experiment and decreased from June to September. Conversely, silage DMI was less for PRE than for POST throughout the experiment and increased over the grazing season. Consequently, total DMI for PRE did not differ from that for POST. Milk urea-N concentration and urinary urea-N excretion in June did not differ between the treatments, whereas that after July was higher for PRE than for POST. Proportion of urinary N excretion to absorbed N intake in June was lower for PRE than for POST, but that after July was higher for PRE than for POST. Feeding NFC supplement before grazing would improve N utilization when cows eat large amounts of herbage high in N.