Suppression of the SOS-inducing activity of mutagenic heterocyclic amine, Trp-P-1, by triterpenoid from Uncaria sinensis in the Salmonella typhimurium TA1535/pSK1002 Umu test

The methanol extract from Uncaria sinensis showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 3-amino-1,4-dimethyl-5H-pyrido[4,3b]indole (Trp-P-1), which requires liver metabolizing enzymes. The methanol extract from U. sinensis was re-extracted with hexane, CH2Cl2, BuOH, and water, respectively. CH2Cl2 extract showed a suppressive effect. A suppressive compound 1 in CH2Cl2 extract was isolated by SiO2 column chromatography. Compound 1 was identified as ursolic acid by IR, electron ionization EI-MS, and NMR spectroscopy. Suppressive effects of ursolic acid (1) and its derivatives, methyl ursolate (1M), acetylursolic acid (1A), and methyl acetylursolate (1MA), were determined in the umu test. These compounds suppressed 61.3, 37.7, 71.5, and 37.8% of the Trp-P-1-induced SOS response at a concentration of 0.4 micromol/mL, respectively. The ID50 values of compounds 1 and 1A were 0.17 and 0.20 micromol/mL. In addition, these compounds were assayed with the activated Trp-P-1. Suppressive effects on activated Trp-P-1 were decreased as compared with those of Trp-P-1.     

Use of gyroscopic sensors for objective evaluation of trimming and shoeing to alter time between heel and toe lift-off at end of the stance phase in horses walking and trotting on a treadmill

OBJECTIVE: To determine whether a shoe with an axialcontoured lateral branch would induce greater lateral roll of the forelimb hoof during the time between heel and toe lift-off at end of the stance phase (breakover). Animals-10 adult horses. PROCEDURE: A gyroscopic transducer was placed on the hoof of the right forelimb and connected to a transmitter. Data on hoof angular velocity were collected as each horse walked and trotted on a treadmill before (treatment 1, no trim-no shoe) and after 2 treatments by a farrier (treatment 2, trim-standard shoe; and treatment 3, trim-contoured shoe). Data were converted to hoof angles by mathematical integration. Breakover duration was divided into 4 segments, and hoof angles in 3 planes (pitch, roll, and yaw) were calculated at the end of each segment. Multivariable ANOVA was performed to detect differences among treatments and gaits. RESULTS: Trimming and shoeing with a shoe with contoured lateral branches induced greater mean lateral roll to the hoof of 3.2 degrees and 2.5 degrees during the first half of breakover when trotting, compared with values for no trim-no shoe and trim-standard shoe, respectively. This effect dissipated during the second half of breakover. When horses walked, lateral roll during breakover was not significantly enhanced by use of this shoe. CONCLUSIONS AND CLINICAL RELEVANCE: A shoe with an axial-contoured lateral branch induced greater lateral roll during breakover in trotting horses, but change in orientation of the hoof was small and limited to the first half of breakover.     

Expression and subcellular localization of GSE protein in germ cells and preimplantation embryos

We previously identified a novel gonad-specific expression gene (Gse) and investigated its expression during gametogenesis in the mouse testis and ovary. In this study, we generated a polyclonal antibody to GSE protein and determined the profiles of the protein's expression in germ cells and preimplantation embryos in detail using immunocytochemical and immunofluorescence staining. In a Western blot analysis, the anti-GSE antibody recognized long and short isoforms (approximately 27.6 kDa and 23.1 kDa) of the protein in the mouse testis and the long isoform in the ovary. In the mouse testis, GSE protein was expressed in spermatocytes I in the pachytene stage, round spermatids, and elongated spermatids. In the mouse ovary, the protein was located in the cytoplasm and nucleus of all oocytes regardless of the stage of the ovarian follicles. In preimplantation embryos from the pronuclear to blastocyst stage, however, GSE protein was mainly detected in the nuclei of cells. At the blastocyst stage, the protein was confirmed to have accumulated in the inner cell mass (ICM), whereas it had mostly disappeared from the trophectoderm (TE). These findings suggest that GSE protein may play a role in the establishment of nuclear totipotency and may be associated with early lineage specification.     

Photodynamic hyperthermal therapy with indocyanine green (ICG) induces apoptosis and cell cycle arrest in B16F10 murine melanoma cells

We examined the effects of photodynamic hyperthemal therapy (PHT), which is a combination of photodynamic therapy (PDT) and hyperthermia (HT), on the apoptosis and cell cycle progression of murine melanoma B16F10 cells. The percentage of apoptotic cell was determined by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI) double staining. The cell cycle analysis was performed by PI staining with flow cytometry. The expression of cyclins and heat shock protein 70 (Hsp70) were examined by a Western blotting analysis. PHT induces death in B16F10 cells, and PHT-mediated apoptosis occurred acutely and persistently in vitro. Our study demonstrated that PHT using indocyanine green (ICG) and near infrared (NIR) light source induces apoptosis and G0/G1 cell cycle arrest in the B16F10 cells.     

Keratometry in normal cats: a cross-sectional study in Japan using an automated handheld keratometer

Keratometry was performed in 73 domestic cats of varied signalment in Japan using an automated handheld keratometer. The mean corneal curvature radius was significantly lower for cats younger than 1 year than for those older than 2 years (8.04 mm vs. 8.80−8.99 mm, P<0.01). The radius was significantly greater in males than in females among the cats older than 11 years (9.22 mm vs. 8.84 mm, P=0.01), while the age distributions of the males and females were similar. Corneal astigmatism did not significantly differ across the gender and age groups. The predictability of the corneal curvature and astigmatism was approximately 41−43% and less than 3%, respectively, as a function of age and bodyweight. The results highlighted some age- and sex-related keratometric variations in domestic cats in Japan.     

Nitrogen uptake by sago palm (Metroxylon sagu Rottb.) in the early growth stages

Previous trials have revealed variable responses of sago palm ( Metroxylon sagu Rottb.) to fertilizer application, particularly nitrogen (N). In the present study, we quantified the fertilizer use efficiency (FUE) of sago palm for the first time using ¹⁵N-labeled fertilizer in pot and field experiments. The pot experiment was conducted in Japan using a 2:1 mixture of sand to Philippine soil. The field experiment was conducted in Leyte in the Philippines. Both experiments consisted of three replicates in each of three treatments: control, ¹⁵N urea at 50 kg N ha⁻¹ and ¹⁵N urea at 100 kg N ha⁻¹. The N uptake of sago palm increased significantly, but inconsistently with increasing N application. The few instances of a significant increase in N uptake did not translate into significant improvements in growth parameters, except for the number of leaflets in the pot experiment. The FUE values for sago seedlings (< 6 months) in the pot experiment treated with 50 and 100 kg N ha⁻¹ were 10.5 and 13.2%, respectively, whereas for the 2-year-old sago palms in the field, the corresponding FUE values were 14.8 and 12.0%. The FUE values were similar at the two levels of N application in both experiments. Sago growth parameters appeared to be insensitive to N application, suggesting that the form of N and the timing of N fertilization are important factors for sago palms. Therefore, the use of N fertilizer in sago production can only be justified after determining and fully understanding the response of sago palm to N application.