Somaclonal variation in Tricyrtis hirta plants regenerated from 1-year-old embryogenic callus cultures

The Liliaceous perennial Tricyrtis hirta, sometimes called ‘Japanese toad lily’, has recently become popular as an ornamental for pot and garden uses. Highly embryogenic callus cultures of this plant predominately consisted of diploid cells but also contained tetraploid cells after 1 year of establishment. In the present study, plans regenerated from the 1-year-old embryogenic callus cultures were subjected to ploidy level analysis and morphological characterization following 3 years of cultivation. Among 37 plants examined, 28 kept the diploid level (2n = 2x = 26) but nine were tetraploid (2n = 4x = 52) as indicated by FCM analysis and chromosome observation. Although no morphological alterations were detected in 26 out of 28 diploid regenerants, the remaining two showed noticeable variations: both were severely dwarf and had crimped leaves and many malformed flowers. The tetraploid regenerants had several horticulturally attractive characteristics compared with the diploid controls, such as longer shoots, thicker stems, and larger flowers. Thus regeneration of tetraploid plants from 1-year-old embryogenic callus cultures offers a possibility to improve the horticultural value of T. hirta, although regeneration of trueness-to-type plants is essential for utilizing the cultures for micropropagation and genetic transformation.     

Responses ofQuercus sapwood to infection with the pathogenic fungus of a new wilt disease vectored by the ambrosia beetlePlatypus quercivorus

Quercus serrata andQ. crispula wilt during the summer in wide areas along the Sea of Japan. Mass attacks of trees by an ambrosia beetle (Platypus quercivorus) are characteristic before appearance of the wilting symptoms. This study investigated the pathogenic effects of a fungus detected specifically in the wilting trees. This hyphomycete fungus,Raffaelea sp., has a distribution that correlates with the discolored xylem area called wound heartwood in which vessels are dysfunctional. Tylosis formation around the hyphae indicates vessel dysfunction. In areas with discoloration, the fungal hyphae were invading living ray parenchyma cells from the vessel lumen. As a protective reaction the ray cells exuded yellow substances into the vessels, but these substances seemed ineffective against the fungal activity, probably because the fungus disperses along the beetle's gallery before enough substance can accumulate. It should allow wide discoloration in sapwood. Cambium was not necrotic around the fungus. The cytological process in the host was as follows: (1) synthesis of secondary metabolites by the stimuli of oak fungus; (2) exudation of yellow substances into vessels; and (3) dysfunction of vessels and wound heartwood formation. In regard to wilting of trees, the pathogenicity of the fungus should be assessed by its ability to stop sap flow.     

The effects of cataract stage, lens-induced uveitis and cataract removal on ERG in dogs with cataract

OBJECTIVE: The purpose of this study was to determine the effects of cataract stage, lens-induced uveitis and cataract removal on the electroretinogram (ERG) of dogs with cataract. ANIMALS STUDIED: Fifty-seven dogs diagnosed with unilateral or bilateral cataract whose ERG was recorded at Rakuno Gakuen University Teaching Animal Hospital from 2001 to 2004. PROCEDURES: Four responses were recorded during the ERG: rod ERG, standard combined ERG, single-flash cone ERG and 30-Hz flicker ERG. Cataracts were divided into four stages: incipient, immature, mature and hypermature, and with or without lens induced uveitis (LIU). Noncataractous eyes of dogs with unilateral cataract were used as the control. We compared ERG amplitude, implicit time, and the b- to a-wave amplitude ratio of cataractous vs. noncataractous eyes, preoperative vs. postoperative cataractous eyes, and cataractous eyes with and without LIU. RESULTS: No significant difference was found in ERG amplitude between incipient, immature and hypermature cataractous eyes, while in mature cataractous eyes decreased amplitude was confirmed in all responses compared with control eyes. However, no significant difference in b/a ratio was found at any stage of cataract. In postoperative eyes, increased amplitude was recorded in all responses compared to preoperative values. In eyes with LIU a decreased amplitude in the rod ERG and b-wave of standard combined ERG was recorded and, furthermore, a significant decline was confirmed in b/a ratio. CONCLUSION: ERG values were influenced by cataract stage and LIU. LIU was associated with a reduction in the b/a ratio.     

Maxillofacial rhabdomyosarcoma in the canine maxillofacial area

Three dogs had a diagnosis of maxillofacial rhabdomyosarcoma. These dogs were treated with surgery and/or radiotherapy, and had poor clinical responses. The tumor tissues in all three cases were observed around the upper premolar teeth with ulcerative lesions and CT examinations in each case revealed extensive bony involvement into the maxilla. Two cases were subjected to surgical excision of the tissues, followed by an external radiation therapy. The other case was only treated with palliative radiation. Outcomes of the treatment of all the cases were quite poor because of the invasive and refractory nature of the tumor cells, leading to the local recurrence and lung metastasis early in the clinical course. All dogs died within two months of the first admission.     

Effect of acetic acid added to cooking water on the dissolution of proteins and activation of protease in rice

The effect of acetic acid on the dissolution of proteins in rice was studied to elucidate the mechanism for the textural change induced by the acid by chemical and SDS-PAGE analyses of the rice proteins in the soaking solution. More proteins were extracted with 0.2 M acetic acid (pH 2.7) than with water (pH 6.8). The effect of acetic acid on the protein dissolution increased with increasing temperature. Immunoblotting confirmed that, when rice was soaked in acetic acid, glutelin was dissolved into the soaking solution and degraded by aspartic proteinase. Aspartic proteinase degraded glutelin much more than it did albumin and globulin. It was found that the combined amount of albumin and globulin dissolved into the acetic acid solution was much larger than that of glutelin, despite the smaller amounts present of albumin and globulin than of glutelin. Metal ions were extracted more with acetic acid than with water. In addition, carboxypeptidase was activated under the acidic condition and resulted in an increase in the amount of free amino acids. The main effect of acetic acid on the dissolution of rice proteins was enhancement of the solubility of albumin, globulin, and glutelin, the effect of proteases being minor.     

Compensatory growth responses in juvenile fat snook,Centropomus parallelus Poey,following food deprivation

The compensatory growth in juvenile fat snook (13.7 g) was evaluated in fish subjected to four treatments: continuously fed (control); subjected to 1 (D1), 2 (D2) and 3 (D3) weeks of food deprivation, followed by a 5‐week re‐feeding period. At the end of food deprivation, the body weights at the different treatments were significantly different and inversely related to the length of the food deprivation period (C=16.425 g; D1=14.024 g; D2=13.542 g; D3=12.228 g). During re‐feeding, no differences were observed in the slopes of body weight growth curves among treatments, indicating failure in full growth compensation. In the first re‐feeding week (week 4), although hyperphagia was observed for all re‐fed groups, better food conversion ratios were detected only for D1 and D2, reflected in an enhancement in the specific growth rates. From week 5 onwards, no differences between treatments and control were detected for any variable. The total food intake was lower for D2 and D3 compared with the control. In the present study, fish showed partial compensatory growth when subjected to a maximum of 2 weeks of food deprivation. Therefore, the food deprivation can promote production losses, even considering a reduction in the total food intake.     

Use of Artemia supplemented with exogenous digestive enzymes as sole live food increased survival and growth during the larviculture of the longsnout seahorse Hippocampus reidi

This study evaluated the effect of exogenous digestive enzymes on the survival, growth performance and morphology of the digestive tract of juveniles of the longsnout seahorse Hippocampus reidi fed exclusively with Artemia and supplemented with five different concentrations of porcine pancreatin (PP) (0, 5, 25, 50 and 75 mg/L from birth until 30 days after release). The results found in the present study clearly show that there was a significant increase in survival and growth as well as substantial changes in the morphology of intestinal villi in seahorses fed exclusively with Artemia supplemented with 75 mg/L PP. Therefore, the use of digestive enzyme supplementation constitutes an important advance for the establishment of a more efficient and practical feeding protocol (exclusive use of Artemia) for juveniles of the longsnout seahorse.     

Rapid and convenient method for preparing aurapten-enriched product from hassaku peel oil: implications for cancer-preventive food additives

Aurapten (7-geranyloxycoumarin) has been reported to be an effective inhibitor of chemical carcinogenesis in some rodent models. In the present study, a method for preparing an aurapten-enriched agricultural product has been established. Out of 17 Rutaceae varieties, the aurapten content in hassaku (Citrus hassaku Hort ex Y. Tanaka) fruit peel was marked, as well as that in natsumikan (C. natsudaidai) and grapefruit (C. paradisi). The aurapten content in hassaku peel was most abundant in April. Hassaku fruit peel oil, which was dissolved by heating precipitates including aurapten which had formed after freezing the peel oil at -20 degrees C, was used. After adsorbing aurapten from peel oil onto synthetic adsorbent SP70, the adsorbent was washed with 40% (v/v) ethanol in water to remove essential oils and pigments remaining on the adsorbent. Aurapten was then eluted with 80% (v/v) ethanol. In a laboratory-scale test, the recovery rates of aurapten and total carotenoids from the eluates were 74.3 and 4.6%, respectively. In a pilot-scale test, the recovery rate of aurapten in the aurapten-enriched preparation from dissolved hassaku oil was 91.0%, and its concentration was 64.1% (w/w). When stored for 180 days under sunlight, aurapten in powder form remained at 88.0-89.0% of the initial level, but only 31.3-43.8% in ethanol. The stability of aurapten in the aurapten-enriched preparation was higher than that of purified aurapten. These results suggest that aurapten is readily recovered from hassaku peel oil using SP70, and thus may be used as a food additive.