A rapid multiple immunofluorescence method for the simultaneous detection of CD20 and CD3 in canine and feline cytological samples

CD20 and CD3 are considered reliable markers for B and T cells, respectively. This study aimed to develop a rapid multiple immunofluorescence (RMIF) method for the detection of CD20 and CD3 on a single cytology slide. Air-dried smears were prepared using samples collected from dogs (n=26) and cats (n=6). Immunosignal detection using the newly developed method required 60 min. Clear immunosignals for CD20 and CD3 were detected in 24 of 26 samples in dogs and in all 6 cats. As the RMIF (CD20/CD3) method can detect markers of both B and T cells simultaneously on a single cytology smear, it would be an efficient tool for the immunophenotyping of canine and feline lymphoma samples.     

Tethered spinal cord related to caudal spinal dysraphism in a tailless Holstein calf

A rare dysraphic caudal spinal anomaly, or caudal agenesis, comprising a tethered spinal cord, was found in a tailless Holstein calf that presented ataxia and paresis with analgesia of the hind limbs. The gently and slimly tapered conus medullaris was poorly formed between S2 and S3 which indicated that it was lying more caudally. The caudal end of the filum terminale adhered to the inner periosteum of the vertebral arch at S4, which is compatible with tethering of the spinal cord. The dysraphic changes from the secondary neurulation error and the longitudinal deranged cord morphology that may have been caused by the caudad traction due to tethering were confirmed. This represents the first bovine case with definitive morphological confirmation.     

Mobility of the forearm skeleton in the Asiatic black (Ursus thibetanus), brown (U. arctos) and polar (U. maritimus) bears

In several primates and carnivores, pronation/supination angles of the forearm skeleton were examined, and it is thought that a larger angle is useful to acquire dexterous behaviors in feeding and/or life style, including climbing. In this study, the pronation/supination angles in Asiatic black, brown and polar bears were nondestructively examined. These specimens were classified as adult or non-adult. Three or four carcasses of each group of Asiatic black and brown bears were used for CT analysis, whereas only one adult polar bear was used. The forearms were positioned within the gantry of a CT scanner in both maximally supinated and pronated states. Extracted cross-sectional CT images of two positions were superimposed by overlapping the outlines of each ulna. The centroids of the radii were detected, and then the centroid of each radius and the midpoint of a line which connects between both ends of the surface of each radius facing the ulna, were connected by lines to measure the angle of rotation as an index of pronation/supination. In adult brown and polar bears, the angles were smaller as compared with the other groups (Asiatic black and non-adult brown bears). Asiatic black and non-adult brown bears can climb trees, whereas adult brown bears and polar bears cannot. This suggests that the pronation/supination angle is related to arboreal activity in Ursidae.     

Maintenance of pregnancy in ovariectomized Mongolian gerbils (Meriones unguiculatus)

Bilateral ovariectomy (Ovx) was carried out on day 20 of pregnancy in Mongolian gerbils ( Meriones unguiculatus ). The body weights of all groups tended to decrease on the day after the operation, and the decrease was significant in the group that was ovariectomized and given vehicle (Ovx + vehicle group). The body weight in this group never recovered until autopsy on day 24, which is normally 1 day before parturition. No fetuses survived to the time of autopsy in any of the animals of the Ovx + vehicle group. Daily administration of 4 mg of progesterone (P₄) prevented the termination of pregnancy in Ovx animals, but 1 mg did not. Treatment with estradiol 17β (E₂) in addition to 4 mg of P₄ tended to result in a lower rate of fetal survival than that of the Ovx group treated with 4 mg of P₄ alone. With regard to fetal weight, treatment with 4 mg of P₄ resulted in the same weight as in the sham-operated controls, but the addition of 0.2 or 1 μg of E₂ to the 4 mg of P₄ resulted in a significantly lower weight than that of fetuses in the 4 mg of P₄ group. The present study suggests that adequate maintenance of pregnancy in ovariectomized gerbils can be achieved by daily treatment with 4 mg of P₄ alone. Moreover, treatment with 0.2 or 1 μg of E₂ in addition to 4 mg of P₄ caused a deterioration in the maintenance of gestation, in contrast to the effects in rats, mice and hamsters.     

Discrimination between Virulent and Attenuated Isolates of Tomato mosaic virus by Restriction Fragment Length Polymorphism

The three missense mutations on the gene for the 130-K protein of Tomato mosaic virus (ToMV) L₁₁A have been thought to be responsible for the attenuation of its virulence. The Eco47I RFLP detecting the missense mutation at 2349 successfully discriminated L₁₁A and its derivative attenuated isolates from ToMV virulent ones. RFLP analysis and mismatch amplification assay detecting the missense mutations at 1117 and 2754, respectively, could not discriminate some of the attenuated isolates from the virulent ones. These results indicated that, of the three missense mutations, only the one at 2349 was conserved in all the L₁₁A-derivative attenuated isolates. Received 16 March 2001/ Accepted in revised form 22 June 2001     

Differences in physiological responses to NaCl between salt-tolerant Sesbania rostrata Brem. & Oberm. and non-tolerant Phaseolus vulgaris L.

Sesbania rostrata ( S. rostrata) Brem. & Oberm., a member of the Fabaceae family, has been used as a promising halophytic plant to ameliorate soil salinity in north-east Thailand. To obtain information regarding the mechanism of salt tolerance, the physiological responses of S. rostrata to NaCl was compared with those of the salt-susceptible species, kidney bean ( Phaseolus vulgaris L. cv. Meal). Seedlings were grown hydroponically with 0, 50, 100 and 150 m m NaCl for 10 days and their effects on growth, chlorophyll content, fluorescence yield ( F _v/ F _m), inorganic elements and amino acid content were determined. The results showed that tolerance to NaCl was clearly different between the two plants. At the highest concentration (150 m m ), the dry weight of S. rostrata was more than 50% greater than the control, whereas the kidney bean could not survive. Chlorophyll a content drastically reduced only in the kidney bean. The F _v/ F _m of S. rostrata did not change with increasing concentrations of NaCl, but that of kidney bean decreased. Greater percentages (≥80%) of absorbed Na⁺ and Cl^– were translocated and accumulated in the shoots of S. rostrata , but remained largely in the roots of kidney bean. The enhancement of contents of amino acids, including proline, with increasing NaCl was observed in both species. These results strongly suggest that the salt tolerance of S. rostrata is associated with the ability of the plant to translocate and sequester Na⁺ and Cl^– in the shoot cells.     

Herbicide pyrazolate causes cessation of carotenoids synthesis in early watergrass by inhibiting 4-hydroxyphenylpyruvate dioxygenase

In aqueous solution, the herbicide pyrazolate [4‐(2,4‐dichlorobenzoyl)‐1,3‐dimethyl‐5‐pyrazolyl p‐toluenesulfonate] is rapidly hydrolyzed to destosyl pyrazolate (DTP), 4‐(2,4‐dichlorobenzoyl)‐1,3‐dimethyl‐5‐hydroxypyrazole, which is an active form of the herbicide. The objective of this study was to examine the effect of pyrazolate and DTP on carotenoids synthesis in susceptible weed, early watergrass (Echinochloa oryzicola Vasing.). Furthermore, their in vitro effect on 4‐hydroxyphenylpyruvate dioxygenase (HPPD) was determined. Roots of the plants at the two‐leaf stage were soaked for 24 h into pyrazolate (5 × 10^–5 mol L^?1) or norflurazon (10^–6 mol L^?1) solution containing 0.5% volume of acetone. At the first sampling time (3 days after treatment: 3 DAT), the chlorophyll content in the third leaves of pyrazolate‐treated plants were not different compared with the untreated control, but it was decreased between 3 and 6 DAT. The declining pattern of β‐carotene in the third leaf of early watergrass was very similar to that of chlorophyll. Both herbicides induced greater accumulation of phytoene in the third leaves of early watergrass 3 DAT, and the levels were kept until 9 DAT. However, feeding of homogentisate reduced the phytoene accumulation only in pyrazolate‐treated plants, suggesting the site of action of the herbicide located in the pathway of plastoquinone synthesis. In a HPPD assay, DTP revealed to inhibit the enzyme with an IC₅₀ value of 13 nmol L^?1 and that of pyrazolate was 52 nmol L^?1. In the pyrazolate solution used in the assay, some of the herbicide possibly has been hydrolyzed to DTP. From the all results obtained, it is strongly suggested that pyrazolate inhibits carotenoids synthesis and causes bleaching on the developing leaves by the similar mechanism with norflurazon, but its action site is not phytoene desaturase and is HPPD.