Real-time PCR genotyping assay for feline erythrocyte pyruvate kinase deficiency and mutant allele frequency in purebred cats in Japan

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.     

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

Effects of food enriched with egg yolk hydrolysate (bone peptide) on bone metabolism in orchidectomized dogs

We examined the effects of chicken egg hydrolysate (also known as “bone peptide” or BP) on bone metabolism in 5- to 8-month-old orchidectomized dogs. The bone formation marker serum bone alkaline phosphatase (BAP) and the bone resorption marker urine deoxypyridinoline (DPD) were used as indicators to measure changes in bone metabolism. The following results were observed that Serum BAP was higher in dogs fed BP-enriched food throughout the clinical investigation. Serum BAP was statistically significantly higher in dogs fed BP-enriched food than in dogs fed non-BP-enriched food at 2 months after orchidectomy. This suggests that BP promoted bone formation immediately after orchidectomy.     

GM1 gangliosidosis in a Japanese domestic cat: a new variant identified in Hokkaido,Japan

A male Japanese domestic cat with retarded growth in Hokkaido, Japan, showed progressive motor dysfunction, such as ataxia starting at 3 months of age and tremors, visual disorder and seizure after 4 months of age. Finally, the cat died of neurological deterioration at 9 months of age. Approximately half of the peripheral blood lymphocytes had multiple abnormal vacuoles. Magnetic resonance imaging showed bisymmetrical hyperintensity in the white matter of the parietal and occipital lobes in the forebrain on T2-weighted and fluid-attenuated inversion recovery images, and mild encephalatrophy of the olfactory bulbs and temporal lobes. The activity of lysosomal acid β-galactosidase in leukocytes was negligible, resulting in the biochemical diagnosis of GM1 gangliosidosis. Histologically, swollen neurons characterized by accumulation of pale, slightly granular cytoplasmic materials were observed throughout the central nervous system. Dysmyelination or demyelination and gemistocytic astrocytosis were observed in the white matter. Ultrastructually, membranous cytoplasmic bodies were detected in the lysosomes of neurons. However, genetic analysis did not identify the c.1448G>C mutation, which is the single known mutation of feline GM1 gangliosidosis, suggesting that the cat was affected with a new variant of the feline disease.     

Blackening of Diospyros genus xylem in connection with boron content

The amount of boron contained in the xylem of Japanese persimmon (Diospyros kaki Thunb.) and ebony (Diospyros ebenum Koen) was determined by inductively coupled plasma (ICP) emission spectrometry and a modified curcumin–acetic acid method. The boron content was compared between the heartwood and sapwood of ebony as well as between a blackened portion (“kurogaki”) and normal portion of Japanese persimmon. The kurogaki contained a higher level of boron than the normal portion of the same individual, although the boron content varied among individuals. Moreover, the boron content of the heartwood of ebony was much higher than that of the sapwood. These results suggest the participation of boron in the blackening of Japanese persimmon. Because both kurogaki and heartwood of ebony are durable to fungal attack, the blackening of Diospyros genus appears to be related to the formation of defensive substances in which boron seems to take part. The convenient curcumin–acetic acid method is an alternative to the ICP method with comparable accuracy.     

Chemical modification of wood by non-formaldehyde cross-linking reagents

Summary Using sulfur dioxide (SO₂) as catalyst, wood specimens were treated with nonformaldehyde cross-linking reagents such as glyoxal, glutaraldehyde and dimethylol dihydroxy ethyleneurea (DMDHEU). The results of dimensional stability, acoustic properties, and mechanical strength tests were compared with those obtained from formaldehyde treatment. With glyoxal and glutaraldehyde treatments, antiswelling efficiency (ASE) reached around 70%, which is comparable to the values attained by formaldehyde treatment, although the accompanying weight gain was much larger than for the latter, whereas DMDHEU did not give sufficiently high ASE. None of the treatments, significantly increased specific dynamic Young's modulus (E/) with the exception of the DMDHEU treatment, loss tangent (tan ) decreased substantially, about 50 and 60% in longitudinal and radial directions respectively, which were somewhat exceeding the formaldehyde treatment. The results suggested that the improvement of the dimensional stability and acoustic properties is partly attributed to the formation of cross-links.     

Dyeability of Chemically Treated Wood and its Discoloration by UV-ray Irradiation

INTRODUCTION Wood is far from a stable material. One of the biggest challenges for woodworking is learning to work within the constraints of wood properties. Discoloration of wood directly influences the decorative performance of wood products. Therefore, the degree of color change is a critical factor that determines the utilization area of dyed wood (Sakuragawa Satoshi, 1996). Discoloration has occurred because the wood components and dye molecules are degraded by UV-ray (Kaneko Shin…     

Stem water transport of Lithocarpus edulis, an evergreen oak with radial-porous wood

The stem water conducting system of an evergreen broad-leaved oak, Lithocarpus edulis (Mak.) Nakai, was investigated. Evergreen broad-leaved oaks (Lithocarpus, Castanopsis, Cyclobalanopsis, Quercus) belonging to the Quercoideae are a major component of Asian monsoon forests, and are characterized by the possession of radial-porous wood. A characteristic of radial-porous wood is the development of aggregate rays between radially oriented files of vessels. We measured the distribution of vessel lumen diameters in a stem cross section and calculated the theoretical water conductivity of the wood. The radial profile of the heat pulse velocity (HPV) was measured for an intact whole tree under field conditions and compared with the theoretical distribution of water conductivity. Soft X-ray photographs of frozen stem sections indicated that most of the vessel lumina were filled with water, including those of vessels more than 20 years old. Even when vessels were relatively wide (lumen diameters > 100 microm), cavitation was negligible. The rate of water uptake from the cut stem base correlated closely with HPV (r = 0.96), and HPV closely reflected the mean volume flow per stem sectional area (SFVS) around the sensor probes. However, the ray tissue sharply inhibited heat transfer, and the positioning of the probes strongly affected the absolute value of HPV. It was also found that HPV more closely reflected the mean sap flow velocity in the vessels than did SFVS.     

Kebon tatangkalan: a disappearing agroforest in the Upper Citarum Watershed,West Java,Indonesia

A comprehensive study was carried out on a typical indigenous agroforestry system, called kebon tatangkalan, in the changing agricultural landscape of the Upper Citarum Watershed, West Java, Indonesia. The main objectives of the study are to elucidate the structural patterns, multidimensional functions, and dynamics of this multi-layered agroforest. The study has identified 12 groups of different plant assemblages indicating that this type of agroforest contributes considerably to the heterogeneity of the agricultural landscape in the study area. Multivariate analysis suggests that elevation and slope are the biophysical factors that correlate most significantly with the distribution of plant species in kebon tatangkalan. The presence of this man-made vegetation is currently declining from the landscape due to population growth and rapid regional economic development. Serious effort to revitalize this traditional agroforest is needed to prevent its disappearance. Introduction of highly productive species and establishment of integrated policies in regional land-use planning and management are measures that should be taken into account in preserving this traditional agroforestry system.     

Application of a modified rate equation to the acetylation of wood

A modified rate equation, in which the diffusion of reagent was taken into consideration, was proposed and applied to results reported previously for the acetylation of wood. The equation was fitted more successfully to the prolonged as well as the early stage of the reaction than a common first-order rate equation. Although the conclusions were virtually unchanged, the reliability was enhanced because reaction parameters such as the rate constant, ultimate extent of the reaction, and activation energy calculated for the modified equation were based on much more data than those reported previously. The equation proposed here may be applicable not only to acetylation but to other chemical modifications of cellulosic materials in heterogeneous systems.