Polyacrylamide gel electrophoresis of S-RNase fragments for identification of S-genotypes of Japanese pear (Pyrus pyrifolia)

Japanese pear (Pyrus pyrifolia) exhibits gametophytic self-incompatibility (GSI) controlled by a complex and multiallelic S locus. The pistil-part product of the S locus is the polymorphic ribonuclease, S-RNase. Information on S-genotypes is important for the production and breeding of Japanese pears. Molecular analyses of S-genotypes of Japanese pear have been conducted with the CAPS (cleaved amplified polymorphic sequence) system; PCR amplification of S-RNase fragments by a common primer pair followed by digestion with restriction enzymes each of which cleaves a specific S haplotype. Here, we show that the separation of S-RNase fragments by polyacrylamide gel electrophoresis (PAGE) distinguishes four out of nine S haplotypes of Japanese pear without restriction digestion. S³-, S⁵-, S⁶- and S⁸-RNases were identified as distinct bands by PAGE. S³- and S⁵-RNases were separated by PAGE despite their identical fragment sizes. Using this system, three Japanese pear lines with unknown S-genotypes were analyzed. The newly determined S-genotypes of the lines were confirmed by CAPS analysis.     

Alteration in the apoptosis process of rat esophageal epithelium with hyperproliferation of indigenous bacteria under a physiological condition

The apoptosis process in rat esophageal epithelium was investigated using enzyme-immunohistochemistry and transmission electron microscopy. As a result, Fas and Fas-L were expressed in the epithelial cell membrane and cytoplasm from the stratum spinosum (SS) to the stratum granulosum (SG). No TNF-R1 show immunopositivity in the cell membranes. TNF-α and caspase-8 were not observed in any layer. Caspase-10, cleaved caspase-3, XIAP and DNase-1 were found in the epithelial cytoplasm from the SS to the SG, whereas Bid, Apaf-1 and cleaved caspase-9 were detected only in the SG. Cytochrome c was observed as cytoplasmic granular positivity from the stratum basale (SB) and altered into homogeneous immunopositivity in the SG. Bcl-2 and Bcl-X immunopositivity was detected in cytoplasm from the SB to the SG. Immunoreactions of Bak in the cytoplasm and Bax beneath the cell membrane were observed from the upper portion of the SS with increasing intensity toward the SG. In the sites with the hyperproliferation of indigenous bacteria, TNF-R1, TNF-α and caspase-8 were detected in the SG and the immunopositive intensities of Bid, Apaf-1 and cleaved caspase-9 were altered to be strong. Prominently swollen cells and decreased mitochondria were ultrastructurally confirmed in the uppermost layers of stratum corneum. These findings suggest that the Fas-Fas-L-interaction initially induces apoptosis through a mitochondria-independent pathway and secondarily through a mitochondria-dependent pathway, leading to eventual epithelial cell death in the rat esophageal epithelium. The bacterial stimuli probably enhance the mitochondria-dependent pathway through the TNF-R1-TNF-α interaction.     

Glycosylated Porphyra-334 and Palythine-Threonine from the Terrestrial Cyanobacterium Nostoc commune

Mycosporine-like amino acids (MAAs) are water-soluble UV-absorbing pigments, and structurally different MAAs have been identified in eukaryotic algae and cyanobacteria. In this study novel glycosylated MAAs were found in the terrestrial cyanobacterium Nostoc commune (N. commune). An MAA with an absorption maximum at 334 nm was identified as a hexose-bound porphyra-334 derivative with a molecular mass of 508 Da. Another MAA with an absorption maximum at 322 nm was identified as a two hexose-bound palythine-threonine derivative with a molecular mass of 612 Da. These purified MAAs have radical scavenging activities in vitro, which suggests multifunctional roles as sunscreens and antioxidants. The 612-Da MAA accounted for approximately 60% of the total MAAs and contributed approximately 20% of the total radical scavenging activities in a water extract, indicating that it is the major water-soluble UV-protectant and radical scavenger component. The hexose-bound porphyra-334 derivative and the glycosylated palythine-threonine derivatives were found in a specific genotype of N. commune, suggesting that glycosylated MAA patterns could be a chemotaxonomic marker for the characterization of the morphologically indistinguishable N. commune. The glycosylation of porphyra-334 and palythine-threonine in N. commune suggests a unique adaptation for terrestrial environments that are drastically fluctuating in comparison to stable aquatic environments.     

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     

Differentiated embryonal rhabdomyosarcoma in a cow.

An embryonal rhabdomyosarcoma was found in the pleura of a 2-year-old Holstein cow after first delivery. The most predominant cells in the tumor were relatively small in size, but considerable numbers of more differentiated cells of larger sizes mingled with the small cells. The most differentiated cells were characterized by multinucleation, abundant cytoplasm containing cross-striated fibrils, intense immunoreactivity for desmin, and weak or negative reactivity for vimentin. Such cells, lacking mitotic activity and displaying weak or no reactivity for proliferating cell nuclear antigen, were considered to be malignant counterparts of myotubes or muscle fibers. This neoplasm seems to follow normal skeletal muscle embryogenesis, and to be capable of differentiation into the final stage of muscle development.     

Comparisons among MRI signs,apparent diffusion coefficient,and fractional anisotropy in dogs with a solitary intracranial meningioma or histiocytic sarcoma

Although MRI has become widely used in small animal practice, little is known about the validity of advanced MRI techniques such as diffusion‐weighted imaging and diffusion tensor imaging. The aim of this retrospective analytical observational study was to investigate the characteristics of diffusion parameters, that is the apparent diffusion coefficient and fractional anisotropy, in dogs with a solitary intracranial meningioma or histiocytic sarcoma. Dogs were included based on the performance of diffusion MRI and histological confirmation. Statistical analyses were performed to compare apparent diffusion coefficient and fractional anisotropy for the two types of tumor in the intra‐ and peritumoral regions. Eleven cases with meningioma and six with histiocytic sarcoma satisfied the inclusion criteria. Significant differences in apparent diffusion coefficient value (× 10^?3 mm²/s) between meningioma vs. histiocytic sarcoma were recognized in intratumoral small (1.07 vs. 0.76) and large (1.04 vs. 0.77) regions of interest, in the peritumoral margin (0.93 vs. 1.08), and in the T2 high region (1.21 vs. 1.41). Significant differences in fractional anisotropy values were found in the peritumoral margin (0.29 vs. 0.24) and the T2 high region (0.24 vs. 0.17). The current study identified differences in measurements of apparent diffusion coefficient and fractional anisotropy for meningioma and histiocytic sarcoma in a small sample of dogs. In addition, we observed that all cases of intracranial histiocytic sarcoma showed leptomeningeal enhancement and/or mass formation invading into the sulci in the contrast study. Future studies are needed to determine the sensitivity of these imaging characteristics for differentiating between these tumor types.     

Modeling of Salt Sorption in Volcanic Ash Soil

Groundwater pollution by nitrate from agricultural fields is a worldwide problem. To improve the understanding of nitrate transport processes through volcanic ash soils overlying groundwater aquifers, salt sorption experiments were performed. Salt sorption is the process that involves the simultaneous use of cation and anion adsorption mechanisms without changing the pH. Prepared six different concentrations of potassium chloride (KCl) solutions were mixed with soil samples. Anion and cation contents of the liquid and solid phases were determined. Experiments were performed in triplicate of six kinds of KCl solutions for three types of soils. Thus, total 54 of samples were analyzed. Relationships between ionic strength and sorption capacity increments of cations and anions were determined. The results show that if ionic strength of the infiltrating solution to the volcanic ash soil is known, the ECEC (effective cation exchange capacity) and EAEC (effective anion exchange capacity) can be reliably estimated. The resulting prediction equations can be included to improve reactive transport simulation models. Consequently, it will be possible to better understand solute transport with changing sorption capacity for the solid soil phase.     

Bartonella species in wild rodents and fleas from them in Japan

The purpose of this study was to assess the role of fleas for transmission of Bartonella species among wild rodents in Japan. Flea samples were collected from wild rodents and examined genetically for Bartonella infection. Bartonella DNA was detected from 16 of 40 (40.0%) flea samples. Sequence analysis demonstrated that 3 of 16 (18.8%) of the Bartonella-positive animals were infested with fleas from which the closely related Bartonella DNA sequence was detected, indicating that the fleas acquired Bartonella from the infested rodents. The DNA was detected in hemolymph, the midgut and the ovary (only in female), indicating that Bartonella might be colonized through the midgut and distributed into the body.     

Long-term survival of a cow with cervical ectopia cordis

This study investigated the long-term survival of a calf with cervical ectopia cordis that grew normally, became pregnant, and calved normally. The cow showed normal cardiac function and absence of peripheral circulation abnormalities. This paper documents antemortem characteristics of the affected cow.     

Application of N2/Ar ratio method for N2 fixation studies in submerged soil

Abstract

Recently there has been developments in the measurement of N₂ fixation due mainly to the C₂H₂ reduction method (1). This method, however, has several disadvantages, especially for submerged soil, and the estimated amount of fixed N₂ on the basis of the C₂H₂ reduction activity is not very reliable. The tracer ¹⁵N₂ technique which gives a reliable estimation of the fixed N₂ is too expensive for common use. Development of an alternative method suitable for submerged soil would therefore be desirable. The present authors expected that the measurement of the ratio N₂/Ar in the soil solution might provide advantages for the estimation of the fixed N₂ in submerged soil.