Detection of γ-H2AX,a Biomarker for DNA Double-strand Breaks,in Urinary Bladders of N -Butyl- N -(4-Hydroxybutyl)-Nitrosamine-Treated Rats

To evaluate the potential role of DNA repair in bladder carcinogenesis, we performed an immunohistochemical analysis of expression of various DNA repair enzymes and γ-H2AX, a high-sensitivity marker of DNA double-strand breaks, in the urothelium of male F344 rats treated with N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN), a bladder-specific carcinogen. Our results clearly demonstrated that γ-H2AX aggregation was specifically generated in nuclei of bladder epithelial cells of BBN-treated rats, which was not found in untreated controls or mesenchymal cells. γ-H2AX-positive cells were detected not only in hyperplastic and neoplastic areas but also in the normal-like urothelium after BBN treatment. These data indicate that γ-H2AX has potential as a useful biomarker for early detection of genotoxicity in the rat urinary bladder. To the best of our knowledge, this is the first report demonstrating expression of γ-H2AX during bladder carcinogenesis.     

Gene dropping analysis of founder contributions in a closed Japanese black cattle population

Gene dropping simulation was applied to Japanese Black cattle population in Hyogo prefecture, to examine the survivals of alleles originated from founder animals. In the analysis, unique alleles were assigned to founders, and the genotypes of all descendants along the actual pedigree were generated through Monte Carlo simulation following Mendelian segregation rules. By replicating this process 10 000 times, the distribution of frequencies of alleles from each founder was estimated. From the distribution, several quantities useful for the management of genetic diversity, such as the probability of allele extinction and the probability of alleles surviving at a critically low frequency were derived. The materials used were 68 781 animals born in 1955–1998 and their pedigree records traced back to the population in 1937 or before. The expected number of alleles retained in the population drastically decreased during the analyzed period, and reached to 57.9 in the population of 1998, which was only 3.3% of the total number of alleles assigned to founders. Detailed analysis of major founders with relatively high genetic contributions to the current population revealed that alleles from most of the major founders are now at high risk of future extinction. These results strongly suggest that for the management of genetic diversity, the genetic contributions of founders are not fully informative, and emphasize the importance of the detection of live animals having founder alleles with high extinction possibilities.     

Emphysematous cystitis in a chemically-induced diabetic dog

Emphysematous cystitis is a rare disorder caused by bacterial infection and characterized by gas accumulation within the bladder wall with cyst formation. This report describes the histopathological characteristics of emphysematous cystitis found in a diabetic female beagle induced by streptozotocin and alloxan. Macroscopically, multiple cyst-like structures were observed on the cut surface of the urinary mucosa. During fixation, small specimens cut from the mucosa floated on the surface of the fixative solution. Histopathologically, multiple cysts were lined with a single layer of flattened cells found to be immunohistochemically positive for vimentin, partially positive for α-smooth muscle actin or macrophage scavenger receptor, class A, and thought to be myofibroblasts, fibroblasts or macrophages. Multinucleated giant cells were observed around the cysts, and gram-negative short bacilli were observed in the lumen of the urinary bladder. From these findings, this case was diagnosed as emphysematous cystitis.     

Hexachlorophene and cuprizone induce the spongy change of the developing rat brain by different mechanisms: the role of 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase)

The goal of this research was to identify mechanisms responsible for the spongy change induced in rats after repeated hexachlorophene (HCP) or cuprizone (CPZ) dosing. Rats were dosed with 35 mg/kg HCP for 5 days followed by drug withdrawal for 7 days suffered spongy changes to the white matter of the cerebrum, cerebellum, medulla oblongata, and spinal cord that were accompanied by degeneration of oligodendroglia. The severity of both lesions increased prominently on day 5; however, the spongy change decreased and degeneration of oligodendroglia reversed on day 12 (7 days after dosing ceased). On day 12, cerebral cortex oligodendroglia were stained strongly by anti-CNPase. Other rats were fed for 8 days with powdered chow containing 1% (w/w) CPZ, which was then withdrawn for 16 days. The rats exhibited the spongy change in the white matter of the cerebrum and cerebellum as well as oligodendroglial cell death from day 3. The severity of both lesions increased prominently on day 8. Cerebral cortex oligodendroglia were stained strongly by anti-CNPase on days 3 to 8 and decreased to the control levels by day 24 (16 days after dosing ceased). Electron microscopy revealed that oligodendroglia frequently displayed apoptotic morphology. These findings suggest that CNPase expression was induced in the course of restoration following HCP-induced insults to oligodendroglia and the myelin sheath, and in the course of demyelination by CPZ-induced damage to oligodendroglia. However, the role of CNPase on both courses is unclear.     

Usefulness of computed tomography for cryptorchidism in bulls

Cryptorchidism is defined as the failure of the testis to descend into the scrotal position. Bulls with cryptorchidism have problems in both meat quality and husbandry management; thus, it is greatly important to accurately identify the retained testis and remove it during the early stage. Abdominal computed tomography (CT) was performed under general anesthesia in 34 bulls aged 3–9 months with cryptorchidism. All bulls underwent laparoscopic or incision approach for cryptorchidectomy, and 40 testes were dissected. The detection rates of retained testes were 64.5% in the abdominal cavity and 100% in the subcutaneous inguinal region, and the total detection rate was 72.5%. Furthermore, all cases in this study were suspected to have intra-abdominal cryptorchidism in primary care, but CT revealed that 22.5% of cases had cryptorchidism in the subcutaneous inguinal region. The CT value (mean ± standard deviation) of the retained testes was 20.96 ± 7.54 Hounsfield Unit, and the CT value and size of the retained testes showed a positive weak correlation with bovine age. Therefore, there is the demerit that general anesthesia and a huge device are necessary; nevertheless, CT is suggested to be useful in identifying the location of retained testes and selecting an appropriate surgical approach in bulls with cryptorchidism. Moreover, CT findings suggested that the maturation of the retained testes might depend not on the descending process but on age.     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

Development of wood and wood ash-based hydroxyapatite composites and their fire-retarding properties

For making efficient use of waste wood ash emitted from wood biomass plant, the wood and wood ash-based hydroxyapatite (HAp) composite was produced and their flammability characterization was studied by thermogravimetric (DTA-TG) analysis, oxygen index (OI) measurement and cone calorimeter test. The results show that the exothermic and weight loss peaks in DTA-TG combustion profiles due to their significant thermal decomposition were weakened by the HAp agent impregnation. In addition, the OI value of HAp composites was increased by the HAp combining and the OI showed a correlation with the HAp contents. Also, the cone calorimeter study revealed that the heat release rates were decreased with increasing amount of HAp injection and accordingly their total heat release has an inverse relationship to the HAp contents. These results indicate that the treatment with wood ash-based HAp agents can enhance the flame retardancy of the treated woods.     

Influence of surface charge on viscosity behavior of cellulose microcrystal suspension

Microcrystalline cellulose with minimal surface charge was prepared from softwood kraft pulp via hydrolysis by 4N hydrochloric acid instead of sulfuric acid. To this material, sulfate ester groups as surface charge were introduced by treating with 55% (w/w) sulfuric acid. A treatment at 60°C for 2h gave nearly the same level of surface charge as that of the H₂SO₄ -hydrolyzed microcrystal. The number of sulfate groups were controlled by changing the H₂SO₄ treatment conditions. Although the microscopic size and shape of the microcrystalline particles were the same irrespective of the preparation method, the introduction of surface charge drastically reduced the viscosity and removed its time dependence. These changes in viscosity behavior are considered to result from charge-induced dispersion of loose aggregates existing in the initial charge-free microcrystals prepared by HCl hydrolysis.Part of this report was presented at the 48th annual meeting of the Japan Wood Research Society, Shizuoka, April 1998