Effects of duration of electric pulse on in vitro development of cloned cat embryos with human artificial chromosome vector

The current applications for cat cloning include production of models for the study of human and animal diseases. This study was conducted to investigate the optimal fusion protocol on in vitro development of transgenic cloned cat embryos by comparing duration of electric pulse. Cat fibroblast cells containing a human artificial chromosome (HAC) vector were used as genetically modified nuclear donor cells. Couplets were fused and activated simultaneously with a single DC pulse of 3.0 kV/cm for either 30 or 60 μs. Low rates of fusion and embryo development to the blastocyst stage were observed in the reconstructed HAC‐transchromosomic embryos, when the duration of fusion was prolonged to 60 μs. In contrast, the prolongation of electric pulse duration improved the embryo development and quality in the reconstructed control embryos without HAC vector. Our results suggested that the optimal parameters of electric pulses for fusion in cat somatic cell nuclear transfer vary among the types used for donor cells.     

Iridal coloboma induces dyscoria during miosis in FLS mice

Objective Fatty liver Shionogi (FLS) mice exhibit characteristic retinochoroidal coloboma because of a failure in fusion of the embryonic optic fissure. However, the same pathogenesis should result in iridal coloboma that has not been reported in this strain. The purpose of this study was to describe the physiologic and morphometric changes in iridal tissue involved in ocular coloboma in FLS mice. Procedures The miotic response after light exposure was evaluated in three strains of live mice, and the shape and location of the pupil were judged macroscopically. Subsequently, macroscopic abnormalities in the anterior segment and fundus were observed postmortem in all mice. During miotic and mydriatic responses in the eyes of live male FLS mice with dyscoric and normal pupils, each iris was measured in four radial directions. The enucleated eyes were examined morphometrically and histologically in both sexes of FLS mice. Results Inferior corectopia upon light‐induced miosis was clearly detected in live FLS mice. The deviated pupils were not round but oval‐shaped. Clinical and postmortem examination revealed that all dyscoric eyes had hypoplastic and dysfunctional irides inferiorly in FLS mice. Histopathological examination confirmed that both the dilator and sphincter muscles and iris stroma were quantitatively diminished in the affected inferior iris. Meanwhile, the rate of fundus (retinochoroidal) coloboma in eyes exhibiting dyscoria was remarkably high, although some dyscoric eyes had no fundus coloboma. Conclusions Fatty liver Shionogi mice had iridal coloboma, resulting in inferior corectopia upon light‐induced miosis as an indicator of ocular coloboma.     

Predicting chronosequential changes in carbon stocks of pachymorph bamboo communities in slash-and-burn agricultural fallow,northern Lao People’s Democratic Republic

In northern Lao People’s Democratic Republic, rising human population has drastically reduced the fallow period of slash-and-burn agriculture which has led to a considerable decrease in the carbon stock in these communities. We estimated chronosequential changes in the communities' carbon stocks, and established the relationship between the fallow period and fallow-period-average carbon stocks in three carbon pools of bamboo-dominated communities in hilly areas of the Luang Prabang Province, northern Lao People’s Democratic Republic. Based on measurements by destructive sampling, we devised a model and root-to-shoot ratios for estimating bamboo biomass. In six secondary plant communities established after slash-and-burn cropping, we estimated community biomass using the above model and others, and measured deadwood and litter stocks. The communities’ biomass and deadwood significantly increased with time after the last cropping and the former reached about 100 Mg ha⁻¹ after 15 years, whereas litter stocks did not show significant trends over time. Extending the fallow period from 2 to 5 years would increase fallow-period-average carbon stock from 14.2 to 25.1 Mg C ha⁻¹. The overstory height was significantly correlated with biomass, deadwood, and litter carbon stocks of these communities. Based on our findings, changes in a community’s carbon stocks can be estimated using the changes in overstory height, which should be taken into account in future studies to reduce uncertainty in estimating carbon stocks in tropical ecosystems.     

Heavy thinning in hinoki plantations in Shikoku (southwestern Japan) has limited effects on recruitment of seedlings of other tree species

Heavy thinning may provide a feasible means to convert single-species coniferous plantations to broadleaved or mixed forests. To assess this possibility, we monitored tree seedling dynamics of other tree species, the understory microenvironment, and seed availability following heavy thinning (50 % by volume) in two hinoki (Chamaecyparis obtusa) plantations in Shikoku, southwestern Japan. Conventional (35 % by volume) and row thinning (50 % by volume) treatments were also applied at one of the sites. Diffuse transmittance in the understory dramatically increased after all thinning treatments then rapidly decreased, except after row thinning. Heavy thinning accelerated recruitment of small seedlings (with stems <30 cm high) at one of the sites. However, the densities of these seedlings declined sharply, and increases in the densities of larger seedlings recorded in subsequent surveys were very small. Furthermore, at the other site, recruitment of small seedlings under heavy thinning was equivalent to that under conventional thinning and lower than that under row thinning. Abundant species in the recruited seedlings were pioneer or subcanopy species found in the soil seed bank. Low seed rain density at both sites appeared to be responsible for the poor recruitment of canopy tree species. Our results suggest that heavy thinning had limited effects on the seedling recruitment of other tree species in these hinoki plantations due to insufficient seed rain and rapid canopy closure.     

Characteristics of systemic infection and host responses in chickens experimentally infected with Salmonella enterica serovar Gallinarum biovar Gallinarum

Salmonella enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) is a host-specific pathogen causing systemic infection in poultry, which leads to significant economic losses due to high mortality. However, little is known about the dynamic process of systemic infection and pathogenic characteristics of S. Gallinarum in chickens. In the present study, we developed an oral infection model that reproduces the pathology of S. Gallinarum and clarified the host immune response of the infected chickens. Chickens at 20 days of age orally inoculated at a dose of 10⁸ colony forming unit (CFU) showed typical clinical signs of fowl typhoid and died between 6 and 10 days post infection. The inoculated S. Gallinarum rapidly disseminated to multple organs and the bacterial counts increased in the liver and spleen at 3 days post infection. Pathological changes associated wirh inflammation in the liver and spleen became apparent at 4 days post infection, and increased expression of interferon (IFN)-γ and interleuikin (IL)-12 in the liver and spleen did not observed until 3 days post infection. These results indicate that S. Gallinarum rapidly spread to entire body through intestine, and the low-level of inflammatory responses in the liver during the early stage of infection may contribute to rapid, systemic dissemination of the bacteria. Our infection model and findings will contribute to the better understanding of the pathogenic mechanism of S. Gallinarum, and provide new insights into the prevention and control of fowl typhoid.     

Antioxidant mechanism studies on ferulic acid: identification of oxidative coupling products from methyl ferulate and linoleate

In our studies of the chain-breaking antioxidant mechanism of natural phenols in food components, ferulic acid, a phenolic acid widely distributed in edible plants, especially grain, was investigated. The radical oxidation reaction of a large amount of ethyl linoleate in the presence of the methyl ester of ferulic acid produced four types of peroxides as radical termination products. The isolation and structure determination of the peroxides revealed that they had tricyclic structures which consisted of ethyl linoleate, methyl ferulate, and molecular oxygen. Based on the formation pathway of the products, a radical scavenging reaction occurred at the 3'-position of the ferulate radical with the four types of peroxyl radicals of ethyl linoleate. The produced peroxides subsequently underwent intramolecular Diels-Alder reaction to afford stable tricyclic peroxides.     

Effects of prescription diet on dealing with stressful situations and performance of anxiety-related behaviors in privately owned anxious dogs

The objectives of this study were to evaluate the effects of a diet (CALM CANINE) supplemented with alpha-casozepine and L-tryptophan on the stress response of anxious dogs with a focus on anxiety-related behavioral parameters and physiological parameters. Forty-four privately owned dogs were first fed with the control diet, followed by the study diet, each for an 8-week period, with a transitional period of 1 week between the diets. After 7 weeks on a particular diet, owners reported their dogs' behavior by filling out a questionnaire. Using the Canine Behavioral Assessment and Research Questionnaire, the effect of the study diet was found to be significant for 4 anxiety-related behavioral parameters, possibly mirroring a placebo effect. After 7 weeks on each diet, the urine cortisol-to-creatinine ratio (UCCR) was measured to assess the stress response to a visit to a veterinary practice for toenail clipping. UCCR was measured to complement the interpretation of behavioral data with regard to the stress susceptibility of the dogs. A positive correlation was found between baseline UCCR, as measured in the home situations, and poststressor UCCR. The stressor-induced increase was significantly lower in the dogs when they were fed the study diet than earlier when they were fed the control diet. The supplementation of casozepine or the proportion of protein in a given diet could have been a determining factor. The study diet seems to improve the ability of an individual to cope with stress and may reduce anxiety-related behavior in anxious dogs.     

Preparation of antibodies directed to the Babesia ovata- or Theileria sergenti-parasitized erythrocytes

To investigate the surface antigens of the bovine red blood cells (RBCs) parasitized by Babesia ovata or Theileria sergenti, attempts were made to produce monoclonal antibodies (mAbs) with BALB/c mice. Comparable numbers of hybridomas producing anti-piroplasm mAbs, as well as anti-bovine RBC mAbs, were obtained from the mice immunized with B. ovata- or T. sergenti-PRBCs. However, mAbs directed to the surface of parasitized RBCs (PRBCs) were obtained only from the mice immunized with B. ovata-PRBCs, but not from those immunized with T. sergenti-PRBCs. When serum samples from the immunized mice and the infected cattle were examined, antibodies recognizing B. ovata-PRBC surface were detected in the sera against B. ovata, but analogous antibodies were undetectable in the sera against T. sergenti, despite that the sera showed substantial antibody titers to T. sergenti piroplasms. The results suggest that significant antigenic modifications occur on the surface of B. ovata-PRBCs, but not on the surface of T. sergenti-PRBCs.