Comparison between immunohistochemistry and genetic clonality analysis for cellular lineage determination in feline lymphomas

Clinical application of genetic clonality analysis by PCR has been available in feline lymphoma. However, it is in under dispute whether the analysis can be applied for cellular lineage determination. To evaluate utility of the analysis for lineage determination, the results between immunohistochemistry and the genetic analysis were compared. Based on the result with immunohistochemistry, sensitivity of the primer for IgH was 89% and specificity was 75%. Sensitivity of the primer for TCRγ was 25% and specificity was 100%. Cross-lineage IgH rearrangement was found in 25% of T-cell lymphomas. The present study suggests that the genetic analysis had better be applied as not a definite but an adjunctive tool for lineage determination in feline lymphomas.     

Ultrastructural and histochemical properties of the olfactory system in the japanese jungle crow, Corvus macrorhynchos

Although it has been commonly believed that birds are more dependent on the vision and audition than the olfaction, recent studies indicate that the olfaction of birds is related to the reproductive, homing, and predatory behaviors. In an attempt to reveal the dependence on the olfactory system in crows, we examined the olfactory system of the Japanese jungle crow (Corvus macrorhynchos) by histological, ultrastructural, and lectin histochemical methods. The olfactory epithelium (OE) of the crow occupied remarkably a small area of the nasal cavity (NC) and had the histological and ultrastructural features like other birds. The olfactory bulb (OB) of the crow was remarkably small and did not possess the olfactory ventricle. The left and right halves of the OB were fused in many cases. In the lectin histochemistry, soybean agglutinin (SBA) and Vicia villosa agglutinin (VVA) stained a small number of the receptor cells (RCs) in the OE and the olfactory nerve layer (ONL) and glomerular layer (GL) on the dorsocaudal region of the OB. Phaseolus vulgaris agglutinin-E (PHA-E) stained several RCs in the OE and the ONL and GL on the ventral region of the OB. These results suggest that 1) the crow has less-developed olfactory system than other birds, and 2) the dedicated olfactory receptor cells project their axons to the specific regions of the OB in the crow.     

In-straw cryoprotectant dilution for bovine embryos vitrified using Cryotop

The aim of this study was to develop an in-straw dilution method suitable for 1-step bovine embryo transfer of vitrified embryos using the Cryotop vitrification-straw dilution (CVSD) method. The development of embryos vitrified using the CVSD method was compared with those of embryos cryopreserved using in-straw vitrification-dilution (ISVD) and conventional slow freezing, outside dilution of straw (SFODS) methods. In Experiment 1, in vitro-produced (IVP) embryos cryopreserved using the CVSD method were diluted, warmed and exposed to the dilution solution at various times. When vitrified IVP embryos were exposed to the dilution solution for 30 min after warming, the rates of embryos developing to the hatched blastocyst stage after 72 h of culture (62.0-72.5%) were significantly lower (P<0.05) than those of embryos exposed to the solution for 5 and 10 min (82.4-94.3%), irrespective of supplementation with 0.3 M sucrose in the dilution solution. In Experiment 2, the rate of embryos developing to the hatching blastocyst stage after 48 h of culture in IVP embryos cryopreserved using the SFODS method (75.0%) was significantly (P<0.05) lower than those of embryos cryopreserved using the CVSD and ISVD methods (93.2 and 97.3%, respectively). In Experiment 3, when in vivo-produced embryos that had been cryopreserved using the CVSD, ISVD and SFODS methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception and delivery rates among groups. In Experiment 4, when IVP embryos derived from oocytes collected by ovum pick-up that had been cryopreserved using the CVSD and ISVD methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception rates among groups. Our results indicate that this simplified regimen of warming and diluting Cryotop-vitrified embryos may enable 1-step bovine embryo transfer without the requirement of a microscope or other laboratory equipment.     

Utility of contrast-enhanced ultrasound in differential diagnosis of adrenal tumors in dogs

This prospective case study aimed to clarify the clinical significance of contrast-enhanced ultrasound (CEUS) for the differential diagnosis of canine adrenal tumors. Forty-three client-owned dogs with adrenal tumors were included. All dogs underwent CEUS, which was evaluated qualitatively and quantitatively. The peak signal intensity (PI), time to peak signal intensity (TPI), mean transit time (MTT), upslope, and downslope were calculated for each time-intensity curve. The histopathological diagnosis of each resected mass was compared with the CEUS findings and parameters. Enhancement distribution, vascularity, tortuous nourishing vessels, enhancement pattern, and late-phase enhancement did not differ significantly between adrenal cortical adenoma (CA), adenocarcinoma (CAC), and pheochromocytoma (PHEO) in qualitative assessment. In PHEO, the TPI was significantly more rapid compared with that in CA (P=0.0287) and CAC (P=0.0404). The MTT in PHEO was significantly shorter than that in CA (P=0.0016) and CAC (P=0.0003). Upslope in PHEO was larger than that in CAC (P=0.0406). Downslope in PHEO was significantly larger than that in CA (P=0.0048) and CAC (P=0.0018). A receiver operating characteristic curve analysis demonstrated that the area under the MTT curve yielded 0.91 for distinguishing PHEO from adrenocortical tumors in dogs; an MTT cut-off value less than 6,225 msec yielded a sensitivity of 69%, specificity of 94%, and likelihood ratio of 12.46. CEUS appears to be clinically applicable for the differential diagnosis between cortical and medullary origins of primary adrenal tumors in dogs.     

Influence of feeding methods on blood biochemical parameters of Japanese Black breeding herd

The aim of this study was to investigate the effect of different feeding methods on blood biochemical parameters by using a total of 32 Japanese Black breeding cows. The diet (silage) was distributed at a rough estimate for 18 days (FN period), and stanchions were not used in this period. After the FN period, the cows were separated in stanchions for feeding and were fed specified amount of silage by using scales on a feeder wagon, which was determined according to the diet formulation and average body weight of cows, for 30 days (FW period). On the last day of the two experiment periods, the body weight of all cows was recorded and blood samples were collected from 10 randomly selected cows. β‐hydroxybutyric acid and albumin (Alb) levels were significantly higher after the FW period than after FN period. The variation of glucose, Alb, calcium and lactic acid was significantly smaller after the FW period. Several blood parameters of the FW period were approximately the appropriate range of a Japanese Black breeding herd in the dry period. Our results suggest that the feeding method by use of a stanchion and proper diet formulation affects blood biochemical parameters and improves nutritional conditions for breeding cows.     

Characterization of Waxy Rice Cakes (Mochi) with Rapid Hardening Quality by Instrumental and Sensory Methods

The textural properties of cooked waxy rice cakes made from four waxy rice varieties including the unique varieties Kantomochi 172 (K172) and BC3 with the property of rapid hardening were analyzed by instrumental and sensory methods. For the instrumental analysis, a compression test, adhesiveness test, and tensile test were conducted. The waxy rice cakes made from K172 and BC3 showed significantly higher compressive force and resistance to break under tensile load. Significant difference in amylopectin chain‐length distribution was observed between each variety, and this difference strongly reflected the hardness of waxy rice cakes. The peak area ratio of amylopectin branch chains with 6–12 degrees of polymerization negatively correlated with the compressive force required for 50 and 80% strain. Sensory evaluation showed that the waxy rice cakes made from these varieties had a significantly harder, less stretchable, less smooth surface, whereas the scores for adhesiveness and ease to cut off (hagire) were preferable to those for other rice varieties.     

Occurrence of Spontaneous Tumors in the Central Nervous System (CNS) of F344 and SD Rats

In order to accurately assess the carcinogenicity of chemicals with regard to rare tumors such as rat CNS tumors, sufficient information about spontaneous tumors are very important. This paper presents the data on the type, incidence and detected age of CNS tumors in F344/DuCrlCrlj (a total of 1363 males and 1363 females) and Crl:CD(SD) rats (a total of 1650 males and 1705 females) collected from in-house background data-collection studies and control groups of carcinogenicity studies at our laboratory, together with those previously reported in F344 and SD rats. The present data on F344/DuCrlCrlj rats (F344 rats) and Crl:CD(SD) rats (SD rats) clarified the following. (1) The incidences of all CNS tumors observed in F344 rats were less than 1%. (2) The incidences of malignant astrocytoma and granular cell tumor were higher in male SD rats than in female SD rats. (3) The incidences of astrocytoma and granular cell tumor were higher in SD rats than in F344 rats. (4) Among astrocytoma, oligodendroglioma and granular cell tumor, oligodendroglioma was detected at the youngest age, followed by astrocytoma, and ultimately, granular cell tumor developed in both strains. The incidences observed in our study were almost consistent with those previously reported in F344 and SD rats.     

Micro‐elevations in paddy fields affect the efficacy of mechanical weeding: Evaluation of weeding machines to control Monochoria vaginalis in herbicide‐free farming

Differences in local topography (micro‐elevation) within a paddy field that constitute a source of variability in agronomical indicators have not been considered thoroughly as a block factor in weed studies. This study investigated and evaluated the performance of weeding machines (weeders) in two herbicide‐free paddy fields that contained micro‐elevations. The plant density of Monochoria vaginalis, a typical and harmful paddy weed unless controlled with herbicides, was used as the indicator of the efficacy of the weeders. Among the three weeders that were tested, one suppressed M. vaginalis dramatically at low elevations and the others were less sensitive to micro‐elevation. For comparison across the fields, micro‐elevations at weed sampling locations were converted to the initial depth of water by using the records of hydrographs that had been set in each field. The relationship between the initial depth of water and the plant density of M. vaginalis was very clear with the use of the elevation‐sensitive weeder. Moreover, this relationship was valid, even with the less‐sensitive weeders. The finding that the greater the depth of water, the less the plant density was significant, even for M. vaginalis, a difficult aquatic paddy weed that was controlled with any of the weeders tested. Thus, micro‐elevation within a paddy field needs to be treated as a crucial block factor in weed‐sampling studies. A coarse survey of the level of a field and the installation of a hydrograph are recommended for a clear analysis of the background of weed control practices.     

Establishment of cell line and in vivo mouse model of canine Langerhans cell histiocytosis

A cell line named FB‐LCH01, derived from a dog diagnosed with Langerhans cell histiocytosis (LCH), was established and characterized. FB‐LCH01 had C‐shaped nucleoli, characterized by modal chromosome aberrations. The original tumour cells as well as established FB‐LCH01 cells were immunopositive for human leukocyte antigen‐DR, Iba‐1 and E‐cadherin, and immunonegative for CD163 and CD204, suggesting Langerhans cell origin. Furthermore, the characteristics of FB‐LCH01 were compared with those of two canine histiocytic sarcoma cell lines (PWC‐HS01 and FCR‐HS02) established previously. Expression of E‐cadherin was detected only in FB‐LCH01, but not in PWC‐HS01 and FCR‐HS02. All (n = 9) the severe combined immunodeficiency mice inoculated with the FB‐LCH01 cells developed subcutaneous tumour masses after 3 weeks. Eight of nine mice also developed metastatic lesions in the lymph nodes (8/8; 100%), lung (5/8; 62.5%), stomach (5/8; 62.5%), heart (4/8; 50%), pancreas (4/8; 50%), kidney (3/8; 37.5%), skin (3/8; 37.5%) and bone marrow (1/8; 12.5%). Tumour cells were pleomorphic and round‐ to polygonal‐shaped with prominent anisocytosis and anisokaryosis. The xenotransplanted tumour cells maintained the immunohistochemical features of the original tumour with persistent E‐cadherin expression at injection site and some visceral organs. In conclusion, the established cell line as well as the mice xenotransplant model in this study reflect the nature of canine LCH and may serve as promising models for investigating the patho‐tumorigenesis and therapy of the disease.