Antigenic variation of porcine transmissible gastroenteritis virus detected by monoclonal antibodies

Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurrence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.     

Adenosine monophosphate degradation and inosinic acid accumulation in the shrimp Penaeus japonicus

International Aquatic Research - Shrimp muscle contains taste component compounds, such as glutamic acid and inosinic acid (IMP). When both are present, taste intensity increases by fourfold to...     

Changes in organosulfur compounds in garlic cloves during storage

We determined the changes in the contents of three gamma-glutamyl peptides and four sulfoxides in garlic cloves during storage at -3, 4, and 23 degrees C for 150 days using a validated high-performance liquid chromatography method that we reported recently. When garlic was stored at 4 degrees C for 150 days, marked conversion of the gamma-glutamyl peptides, gamma-L-glutamyl-S-allyl-L-cysteine and gamma-L-glutamyl-S-(trans-1-propenyl)-L-cysteine (GSPC), to sulfoxides, alliin and isoalliin, was observed. Interestingly, however, when garlic was stored at 23 degrees C, a decrease in GSPC and a marked increase in cycloalliin, rather than isoalliin, occurred. To elucidate in detail the mechanism involved, the conversion of isoalliin to cycloalliin in both buffer solutions (pH 4.6, 5.5, and 6.5) and garlic cloves at 25 and 35 degrees C was examined. Decreases in the concentration of isoalliin in both the solutions and the garlic cloves during storage followed first-order kinetics and coincided with the conversion of cycloalliin. Our data indicated that isoalliin produced enzymatically from GSPC is chemically converted to cycloalliin and that the cycloalliin content of garlic cloves increases during storage at higher temperature. These data may be useful for controlling the quality and biological activities of garlic and its preparations.     

The relation between pathogenicity and plaque size of Getah virus Kanagawa strain in suckling mice

The relation among biological properties, particularly pathogenicity for suckling mice, and plaque size was studied in four virus strains: Getah virus strain Kanagawa; two strains obtained by plaque cloning of the Kanagawa strain, Getah Kanagawa SP (G-K-SP) strain which forms small plaques (SP) only and strain G-K-LP which forms large plaques (LP) only; and strain Haruna which forms SP only. There were no marked differences among the four strains in serological properties, growth curves and sensitivity to pH, trypsin and temperature. Strain G-K-LP showed higher pathogenicity for suckling mice than strain G-K-SP. However, the pathogenicity of strain Haruna, which forms SP only, was as high as that of strain G-K-LP. Some of the clones in SP of strain Kanagawa kill all mice in 5 to 6 days after inoculation while the others required 9 to 11 days or longer before causing death. The present study showed that the pathogenicity of Getah viruses shortly after being isolated from the field, such as the Kanagawa strain, is different between large and small plaques, and even among small plaques, at least in suckling mice, and that the pathogenicity has no relation to plaque size.     

Thyroid Hormone-disrupting Effects and the Amphibian Metamorphosis Assay

There are continued concerns about endocrine-disrupting chemical effects, and appropriate vertebrate models for assessment of risk are a high priority. Frog tadpoles are very sensitive to environmental substances because of their habitat and the complex processes of metamorphosis regulated by the endocrine system, mainly thyroid hormones. During metamorphosis, marked alteration in hormonal factors occurs, as well as dramatic structural and functional changes in larval tissues. There are a variety of mechanisms determining thyroid hormone balance or disruption directly or indirectly. Direct-acting agents can cause changes in thyroxine synthesis and/or secretion in thyroid through effects on peroxidases, thyroidal iodide uptake, deiodinase, and proteolysis. At the same time, indirect action may result from biochemical processes such as sulfation, deiodination and glucuronidation. Because their potential to disrupt thyroid hormones has been identified as an important consideration for the regulation of chemicals, the OECD and the EPA have each established guidelines that make use of larval African clawed frogs (Xenopus laevis) and frog metamorphosis for screening and testing of potential endocrine disrupters. The guidelines are based on evaluation of alteration in the hypothalamic-pituitary-thyroid axis. One of the primary endpoints is thyroid gland histopathology. Others are mortality, developmental stage, hind limb length, snout-vent length and wet body weight. Regarding histopathological features, the guidelines include core criteria and additional qualitative parameters along with grading. Taking into account the difficulties in evaluating amphibian thyroid glands, which change continuously throughout metamorphosis, histopathological examination has been shown to be a very sensitive approach.     

Modified plant architecture to enhance crop disease control: genetic control and possible value of upright fruit position in cucumber

Plant architectural features can facilitate disease control by creating unfavourable environments for pathogen growth or limiting pathogen contact with the host. In the case of Phytophthora capsici infection of cucumber (Cucumis sativus), the susceptible fruit typically lie in contact with soil under warm, moist conditions of a full canopy, an ideal location for exposure to inoculum and disease development. We have shown that increased row spacing, trellising and architectural variants that allow for more open canopies, such as shorter vines, reduced branching, or smaller leaves, can modify microclimate as assessed by temperature at the location of developing fruits. However, only trellising reduced infection rates (<3 % infection vs. 22 % for control). A cucumber accession PI308916, with extremely short internodes and resulting upright position of young fruit, also exhibited reduced disease. Fruit of PI308916 were susceptible when inoculated with P. capsici, indicating that reduced infection likely resulted from decreased soil contact. The compact trait is inherited as a single recessive gene, cp. Like many architectural traits, cp may be pleiotropic; PI308916-derived breeding lines were reported to have poor seedling emergence. PI308916 exhibited inconsistent apical hook formation that co-segregated with short internodes in F₂ and backcross generations, and disorganized internodal cortical cell arrangement. The combined phenotypes suggest either pleiotropy or very tight linkage of genes affecting hormone balance or cell division. The narrow chromosome region recently defined by QTL analysis includes candidate gene homologs of meristem related genes modulating cell division or spacing of lateral shoot organs and genes associated with brassinosteroid signaling or cytokinin content.     

Spatial distribution pattern of attack of the oak borer,Platypus quercivorus (Murayama) (Coleoptera: Platypodidae), and scolytid ambrosia beetles (Coleoptera: Scolytidae) on fresh logs

The spatial distribution patterns of the attack on fresh logs ofPasania edulis was studied for the oak borer,Platypus quercivorus (Murayama), and two species of Scolytid ambrosia beetles,Xylosandrus crassiusculus (Motschulsky) andXyleborus attenuatus Blanford, in 1994 and 1995. On the logs where onlyP. quercivorus attacked, the entry holes were distributed uniformly when attack intensity was low. However, the distribution pattern became more aggregated with the increase in attack intensity. On logs where bothP. quercivorus and the two Scolytid species attacked, there was a negative association between the spatial distribution of the entry holes ofP. quercivorus and that of scolytids. Simultaneous attack of two scolytids also increased the degree of aggregation of the entry holes ofP. quercivorus. The entry holes of scolytids were distributed in groups irrespective of the attack intensity of scolytids andP. quercivorus. These results suggest an asymmetrical interspecific relationship betweenP. quercivorus and scolytids. Concentration of the entry holes ofP. quercivorus in a small area may cause a considerable decline in the reproductive success in the galleries constructed there.     

Plantlet regeneration from leaf protoplasts ofPopulus euphratica

Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the cell density of 9×10⁴/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of 2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and plantlets were regenerated on 1/2 MS medium without a hormone. A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995).