Development of genomic and EST-SSR markers in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.) belongs to Brassicaceae family and is a close relative of Brassica. This species shows a wide morphological diversity, and is an important vegetable especially in Asia. However, molecular research of radish is behind compared to that of Brassica. For example, reports on SSR (simple sequence repeat) markers are limited. Here, we designed 417 radish SSR markers from SSR-enriched genomic libraries and the cDNA data. Of the 256 SSR markers succeeded in PCR, 130 showed clear polymorphisms between two radish lines; a rat-tail radish and a Japanese cultivar, ‘Harufuku’. As a test case for evaluation of the present SSRs, we conducted two studies. First, we selected 16 SSRs to calculate polymorphism information contents (PICs) using 16 radish cultivars and four other Brassicaceae species. These markers detected 3–15 alleles (average = 9.6). PIC values ranged from 0.54 to 0.92 (average = 0.78). Second, part of the present SSRs were tested for mapping using our previously-examined mapping population. The map spanned 672.7 cM with nine linkage groups (LGs). The 21 radish SSR markers were distributed throughout the LGs. The SSR markers developed here would be informative and useful for genetic analysis in radish and its related species.     

In vitro and in vivo comparison of applanation tonometry and rebound tonometry in dogs

Intraocular pressure (IOP) evaluated by applanation tonometry via TONO-PEN XL (TP), and rebound tonometry via TonoVet (TV) were compared in enucleated canine eyes with varied pressure of the anterior chamber (AC) and in clinical cases. TV measured IOP values were lower than IOP measurements of TP in the enucleated eyes with 5-10 mmHg of AC (P<0.0001), though there was no significant difference in IOP values obtained with TP and TV on the pressure ranges of 15-20 mmHg. However, TP detected IOP values were lower than IOP measurements of TV in the eyes with over 25 mmHg of AC (P<0.0001). The results of clinical cases were similar to the enucleated eye model. There was no significant difference in IOP values obtained from TP and TV in dogs with normotensive eyes. IOP measurements of TP were lower than those of TV in glaucomatous eyes (P<0.0001). TV was a reliable tonometer for measurement of IOP in hypertensive eyes, whereas it was less accurate than TP in hypotensive eyes. The characteristics of TP and TV should be considered in the evaluation of IOP in practice.     

The effect of ovarian status and follicular diameter on maturational ability of domestic cat oocytes

The objective of this study was to clarify the effect of ovarian status and follicular size on morphological normality and maturational ability of cat oocytes. Ovarian status was classified into inactive, follicular, luteal and prepubertal, and follicles were classified into three groups according to their diameter (400-800, 800-1200 and 1200-2000 μm). In each ovarian status, the number of follicles decreased but the percentage of morphologically normal oocytes increased with the growth of follicles (p<0.05). Only a single follicle that was 1200-2000 μm in diameter was observed in two of the five prepubertal cats. In follicles that were 800-1200 μm in diameter, the percentage of normal oocytes and maturation rate were higher in prepubertal cats than in mature cats (p<0.05). Oocyte diameter tended to increase with the growth of follicles. After oocytes were cultured individually in droplets of maturation medium, the oocyte maturation rate increased with the growth of follicles in each ovarian status (p<0.05). In conclusion, oocytes collected from larger follicles possess higher maturational ability in vitro in sexually mature cats. In prepubertal cats, a higher maturation rate can be obtained from oocytes derived from small follicles compared with in mature cats.     

Cerebral vascular hamartoma with thrombosis in a dog

A cerebral vascular hamartoma was identified in the frontal lobe, striatum and thalamus of the right side of the brain of a male, 7-year-old Shih Tzu. Histologically, the lesion consisted of thin-walled vessels, which showed various sizes and occasionally contained fibrin thrombi. These vascular walls were composed of a single layer of fibromuscular tissue lined by flat endothelium with various amount of collagen, but devoid of large coat of smooth muscles and elastic tissue. Immunohistochemically, the lining endothelial cells were positive for von Willebrand Factor antibody. Neuropil between the vessels was stained with Klüver-Barrera stain, and positive for synaptophysin and GFAP antibodies. Based on these findings, the lesion was diagnosed as vascular hamartoma, which might resemble venous malformation in humans.     

Amelanotic melanocytic tumors of the pinna in six F344 rats.

Spontaneous amelanotic melanocytic tumors of the pinna were found in six females of 960 male and 960 female albino (F344/DuCrj) rats which had been used in three different 24-month chronic toxicity studies. The age when the pinnal tumors were detected ranged from 37 to 59 weeks. The tumors were located unilaterally in the pinna and observed as subcutaneous spherical to irregular, solid white masses measuring 7 to 25 mm in diameter. The pinnal tumors were histologically classified into spindle cell and pleomorphic cell types. The spindle cell type was observed in four rats and composed of fusiform cells arranged in interlacing bundles. The pleomorphic cell type was observed in the remaining two rats and composed of pleomorphic large cells arranged in sheets. One tumor of the latter type metastasized to the submaxillary lymph node and lung. Melanin pigments were not demonstrated in any of the tumors. In immunohistochemistry, nuclei and cytoplasm of tumor cells in all the tumors were slightly positive for S-100 protein. Ultrastructurally, tumor cells contained a considerable number of premelanosomes in the cytoplasm. Desmosomes were occasionally observed between the cell membranes of the adjacent tumor cells. No distinct basal lamina was seen around tumor cells.     

Proposal of new return flow analysis by replacement-in-order method for paddy irrigation water

Return flow and repeated use of irrigation water for paddies is the most important issue in the Asian monsoon region, because sometimes this water is applied in greater quantity than that of evapotranspiration plus percolation. A new return flow analysis, the “replacement-in-order method”, which introduces a unique numbering system for very complicated irrigation and drainage networks, is proposed for the main canal with the dual purposes of irrigation and drainage. The method is applied to the Shichika irrigation district in the ordinal (season) irrigation period, resulting in a return flow ratio of 45 % for the entire area. Of this amount, 25 % is available for irrigation again. The remaining 20 % is unavailable, because the return flow discharged directly into a canal lacking a diversion weir in the drainage system, or into the Japan Sea. The return flow ratio is very different at the main canal location, from no return flow to 88 %. With the aid of the above method, theoretical analysis of return flow for paddy irrigation water can be done. This includes the deterministic return flow ratio inside and outside the irrigation area, plus precise information of return flow ratios at various main canal locations and routes of irrigation and drainage water.