Survey of tick infestation and tick-borne ehrlichial infection of dogs in Ishigaki Island, Japan.

Twelve (54.5%) of 22 free-roaming dogs in Ishigaki Island had tick infestation identified as Rhipicephalus sanguineus. There were 121 ticks recovered and consisted of 28 females, 58 males, 22 nymphs and 3 larvae. Infection of dogs possibly with canine ehrlichial pathogens was examined by both indirect immunofluorescence assay and polymerase chain reaction (PCR). Two dogs of the 13 examined were sero-positive for the human granulocytic ehrlichia agent, and one of two dogs was PCR positive for Ehrlichia platys. This dog had platelet numbers slightly lower than normal value, however, no morulae were found within platelet on peripheral blood smear stained with Giemsa.     

Body and intestinal growth of broiler chicks on a commercial starter diet. 2. Development and characteristics of intestinal enzymes

1. Investigations were conducted into the development of intestinal enzyme function in broiler chickens on a commercial starter diet. The differences between intestinal regions and localisation of enzymes on the villus were assessed. 2. The specific activity of maltase, sucrase, aminopeptidase N (APN) and alkaline phosphatase (AP) at all intestinal sites decreased with age. There were also variations between intestinal sites although this variation depended on age. The specific activity of maltase was higher than that of the other enzymes examined, regardless of age and intestinal site. The total activities of the enzymes also increased with age at all intestinal sites. 3. Results of the localisation of enzymes on the crypt: villus axis showed that activity was expressed over a large proportion of the villus. There was an increase in the total villus activity of alpha-glucosidase (AG), APN and AP with age. Activity per unit villus surface area was similar between ages, except for jejunal AP. At hatch enzyme activity was expressed over 44.1, 55.8 and 63.3% of villus height in the duodenum, jejunum and ileum, respectively. At 21 d of age, corresponding values were 68.7, 65.6 and 77.2%. The point of peak activity from the crypt: villus junction increased with age. In the jejunum, most enterocytes were capable of secreting active enzymes within 1 h of formation. Cells maintained their secretory capabilities until they were more than 60 h old in the case of AG. 4. Although the specific activities of the enzymes were maximal at hatch, the digestive capacity of older birds may be sustained by an increase in total enzyme activity brought about by increased surface area. The pattern of enzyme activity along the gastrointestional tract (GIT) and crypt: villus axis is similar to that reported for some mammalian species.     

Effects of salivary gland extract from Rhipicephalus sanguineus on IgG subclass production and cytokine mRNA expression in mononuclear cells of canine peripheral blood

The effects of salivary gland extract (SGE) from R. sanguineus were examined on the production of IgG1 and IgG2 and the mRNA expression of IFN-gamma, IL-2, IL-4, IL-5 and IL-10 in the mononuclear cells from canine peripheral blood, treated with concanavalin A (ConA) in vitro. SGE suppressed the ConA-induced production of IgG2. It also inhibited the expression of IFN-gamma, IL-2 and IL-5 mRNA in a dose-dependent manner. No dose-dependent suppression was observed of IL-10 mRNA expression although a significant effect was observed at a SGE protein concentration of 25 microg/ml. SGE had no effect on the mRNA expression of IL-4. These results suggest that the suppression of IgG2 production by SGE from R. sanguineus was caused by the suppression of IFN-gamma production.     

Ceramide aminoethylphosphonate from jumbo flying squid Dosidicus gigas attenuates the toxicity of cyanotoxin microcystin-LR

We have previously established the method for isolation of ceramide aminoethylphosphonate (CAEP) from jumbo flying squid Dosidicus gigas. In this study, we performed a MTT assay to evaluate the safety of CAEP to the cell lines for the application to health food and supplements. The CAEP did not show any cytotoxicity to various HEK293-transfectant cells. Next, we elucidated the positive function of CAEP to the somatic cells. Recently, we have reported that hepatotoxin microcystin-LR was taken up into the hepatocytes mediated by hepatocellular uptake transporters OATP1B1 and OATP1B3, and the cells were induced cytotoxicity subsequently. Cytotoxicity of microcystin-LR to permanently OATP1B3-expressing HEK293-OATP1B3 cells rather than to HEK293-OATP1B1 cells was preferentially attenuated by CAEP in a concentration-dependent manner. In addition, the enzyme activity of serine/threonine phosphatase, which was inhibited by microcystin-LR, was recuperated by co-exposure to CAEP. Furthermore, microcystin-LR-induced cellular protein phosphorylation were disrupted by CAEP exposure. These results suggested that CAEP is a promising remedy and/or preventive medicine for liver damage with microcystin-LR.     

Intestinal function and reproductive capacity of Tegel pullets in response to exogenous oestrogen.

The effects of varying levels of exogenous oestrogen (E2) (0, 10 or 100 micrograms E2/kg BW) on the development of 18-week old pullets were tested over a 28-day period. The hormone had no significant effects on feed intake, body growth, feed conversion ratio or weight of the oviduct. Similarly, there were no significant effects of the hormone on egg production and egg weight but eggshell thickness and weight of shell per unit area were increased (P < 0.05) at a lower level of administration (10 micrograms E2/kg BW), compared to the control and the highest level of hormone. The morphometry of the jejunal mucosa and some enzymes associated with Ca transport were similar between the three groups. Oestrogen treatment, however, intensely enhanced the expression of calbindin D22K, although this was not quantified.     

Variation and Segregation for Rachis Fragility in Spelt Wheat, Triticum Spelta L.

Spelt wheat, Triticum spelta L., has been proved to be rich-sources of useful genes for tolerance to biotic and abiotic stress, and grain quality. But this crop plant has some undesirable traits including glume tenacity and brittle rachis. Free-threshing and reduced fragility of rachis are very important traits for cultivation. The objectives in the present study were to investigate genetic variation of rachis fragility in a wide range of spelt accessions, to examine its genetic segregation pattern, and to clarify if rachis fragility is associated with dosage of chromosome 5A in aneuploid lines of bread wheat. The results demonstrated that spelt germplasm contains a wide range of rachis toughness, and thus selection of spelt wheat with desirable characteristics combined with an appropriate level of tough rachis would be possible. Spike morphology in the F₂ plants was segregated into the three types, square-headed, speltoid, and compactoid. The F₂ plants with compactoid spikes had the most brittle rachis, followed by the speltoid and square-headed spike F₂ plants. Rachis fragility in bread wheat also had genetic variation and was associated with dosage of chromosome 5A.     

CTLA-4 control over Foxp3+ regulatory T cell function

Naturally occurring Foxp3+CD4+ regulatory T cells (Tregs) are essential for maintaining immunological self-tolerance and immune homeostasis. Here, we show that a specific deficiency of cytotoxic T lymphocyte antigen 4 (CTLA-4) in Tregs results in spontaneous development of systemic lymphoproliferation, fatal T cell-mediated autoimmune disease, and hyperproduction of immunoglobulin E in mice, and it also produces potent tumor immunity. Treg-specific CTLA-4 deficiency impairs in vivo and in vitro suppressive function of Tregs-in particular, Treg-mediated down-regulation of CD80 and CD86 expression on dendritic cells. Thus, natural Tregs may critically require CTLA-4 to suppress immune responses by affecting the potency of antigen-presenting cells to activate other T cells.     

Immunohistochemical study on the delayed progression of epithelial apoptosis in follicle-associated epithelium of rat Peyer's patch

It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi.