Prophylactic efficacy of desmopressin acetate for diabetes insipidus after hypophysectomy in the dog

The prophylactic efficacy of desmopressin acetate (DDAVP) on diabetes insipidus (DI) after hypophysectomy was investigated in the dog. In the control group, hypernatremia with a plasma level of 155 mEq/l or higher persisted for 12 hr from the 4th to the 16th hour after hypophysectomy, and symptoms of DI developed within five days after surgery. In the DDAVP treatment group, these changes were not observed, showing that administration of DDAVP (4 microg, installation, twice daily) effectively prevented hypernatremia that develops immediately after surgery and DI-like symptoms that persists for about one week after surgery.     

The relationship between cellular radiosensitivity and radiation-induced DNA damage measured by the comet assay

The relationship between deoxyribonucleic acid (DNA) damage and the cell death induced by gamma-irradiation was examined in three kinds of cells, Chinese hamster ovary fibroblast CHO-K1, human melanoma HMV-II and mouse leukemia L5178Y. Cell survival was determined by a clonogenic assay. The induction and rejoining of DNA strand breaks induced by radiation were measured by the alkaline and neutral comet assays. L5178Y cells were the most radiosensitive, while CHO-K1 cells and HMV-II cells were radioresistant. There was an inverse relationship between the survival fraction at 2 Gy (SF2) and the yield of initial DNA strand breaks per unit dose under the alkaline condition for the comet assay, and also a relationship between SF2 and the residual DNA strand breaks (for 4 hr after irradiation) under the neutral condition for the comet assay, the latter being generally considered to be relative to cellular radiosensitivity. In the present analysis, it was considered that the alkaline condition for the comet assay was optimal for evaluating the initial DNA strand breaks, while the neutral condition was optimal for evaluating the residual DNA strand breaks. Since the comet assay is simpler and more rapid than other methods for detecting radiation-induced DNA damage, this assay appears to be a useful predictive assay for evaluating cellular clonogenic radiosensitivity of tumor cells.     

cDNA cloning of a Rab5 homologue in Babesia gibsoni

For studying protein trafficking in Babesia-infected erythrocyte, we describe the cloning of a Rab5, one of molecular marker for vesicle trafficking in eukaryotic cells, gene homologue in Babesia gibsoni (BgRab5). The full-length cDNA of BgRab5 is 1,020 bp long with an open reading frame encoding a protein of 220 amino acids. The deduced amino acid sequence of BgRab5 contained the highly conserved GTP-binding consensus sequence and shares about 40% homology with that of Rab5 from Plasmodium falciparum, Toxoplasma gondii, Dog, Lotus japonicusor, Oryza sativa. Northern blot analysis showed that the BgRab5 probe hybridized with a 1kb band in total RNA from parasitized erythrocytes, that was consistent with the size of the BgRab5 full-length cDNA.     

Molecular cloning of canine membrane-anchored inhibitor of matrix metalloproteinase, RECK

The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene is one of the endogenous matrix metalloproteinase (MMP) inhibitors. It was reported that decreased RECK expression closely correlated with tumor malignancy. We determined the cDNA sequence of the canine RECK gene. The cDNA sequence and deduced amino acid of canine RECK were 2,913 bases and 971 residues, respectively. The predicted amino acid sequence of the protein showed 95.5% and 91.9% homology with human and mouse RECK, respectively. RECK mRNA expression was analyzed in various canine tissues and tumor cell lines by quantitative RT-PCR. The highest RECK expression was detected in lung and testis. In comparison with the tissues, a remarkably low expression level was detected in tumor cell lines. In addition, the RECK gene was transfected in the canine transitional cell carcinoma, and its influence on cell proliferation, migration, and invasion was analyzed. The transfected RECK gene suppressed only canine tumor invasion. These results showed that RECK might play an important role in tumor malignancy in dogs as well as in other mammalians.     

Antibabesial activity of protoberberine alkaloids and 20-hydroxyecdysone from Arcangelisia flava against Babesia gibsoni in culture

Bioassay-guided fractionation of the boiled extract from the stems of Arcangelisia flava led to the isolation of palmatine (1), berberine (2), jatrorrhizine (3), dihydroberberine (4) and 20-hydroxyecdysone (5). The chemical structures of these compounds were elucidated on the basis of their chemical and spectral evidence. The isolated compounds were evaluated for their growth inhibiting effects on Babesia gibsoni in culture for a week. Compounds (1-4) showed significant inhibitions at concentrations from 100 to 1.0 microg/ml, while compound 5 at a concentration of 100 microg/ml, only.     

Ultrasonographic monitoring of nuclear transferred fetal weight during the final stage of gestation in Holstein cows

Dystocia or stillbirth accompanied by Large Offspring Syndrome (LOS) occurs rather frequently in Holstein nuclear transferred calves. In regard to prophylaxes, nuclear transferred Holstein fetuses were monitored with ultrasonography during the final stage of gestation. Fetal weight was estimated weekly based on the fetal metacarpal width using ultrasonography. Fourteen Holstein cows pregnant with Holstein nuclear transferred fetuses were the subjects of this experiment. The fetal weight was estimated by measuring the fetal metacarpal width during the last month of gestation according to the expected date of parturition. Measurements were performed on a weekly basis. The ultrasound-estimated metacarpal width and body weight of 13 of the fetuses in the last week of gestation (30.2+/-2.2 mm, 50.0+/-4.7 kg) were similar to the actual measurement immediately after birth (30.0+/-2.1 mm, 51.2+/-5.5 kg). These results indicate that ultrasonographic monitoring within a week of parturition to is accurate for estimating fetal weight. Prediction of LOS with ultrasonography contributes to reliable a diagnostic method that minimizes syndrome-related gyneco-obstetric complications at parturition with the aid of appropriate treatments.     

Morphological and cytological characteristics of protoplast-derived plants of statice (Limonium perezii Hubbard)

Morphological characteristics of 56 protoplast-derived plants (protoclones) of statice (Limonium perezii Hubbard) were compared with meristem-derived plants (controls). Some protoclones had smaller leaf shape index (length/width) and shorter length of flower stalk and calyx than the controls. However, no distinct somaclonal variations were observed in other characteristics such as color of calyx and number of petals and stamens, except for protoclone No. 1–7, which had flowers with abnormal petals and low pollen fertility (25%). Microscopic observation showed that all of the 17 protoclones examined, including clone No. 1–7, had the normal diploid chromosome number of 14.     

Regeneration of cartilage tissue by autologous chondrocytes transplantation for cartilage defects in a experimental bovine model

To evaluate the effects of chondrocytes transplantation on the regeneration of cartilage by intraarticular injection or injection into blood clots at cartilage defects, eight full-thickness cartilage defects were created surgically on the articular surface of each femoral trochlea of two calves. Autologous chondrocytes were isolated individually from the cartilage pieces collected at the creation of defects. And isolated cells were cultured in monolayers for proliferation. Cells were injected into synovial fluid (Group 2, n=11) or into the blood clots at the cartilage defects (Group 3, n=5) of the left femoropatellar joint on weeks 2 and 3, respectively after the operation. The defects (Group 1, n=16) of right femoropatellar joint were left untreated in the control group. After 14 weeks, repaired tissues were evaluated based on gross and histological examinations. In Group 3, more repaired tissues and a better interface between the repaired tissue and host cartilage were observed compared with the results for Groups 1 and 2. Moreover, cartilaginous tissue were observed more in defects of Group 3 than in defects of other groups. In conclusion, the present study suggests that the injection of cells into the blood clot at a cartilage defect might be applicable for the regeneration of damaged cartilage.     

The TIR–NBS but not LRR domains of two novel N-like proteins are functionally competent to induce the elicitor p50-dependent hypersensitive response

The tobacco N protein recognizes the helicase domain (p50) of the Tobacco mosaic virus (TMV) replicase as an elicitor and mediates hypersensitive response (HR). We obtained two cDNA clones encoding novel N-like (NL) proteins NL-C26 and NL-B69 from Nicotiana tabacum cv. Samsun NN. NL-C26 and NL-B69 had a Toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat (TIR–NBS–LRR) structure and showed 78% and 73% identities to N, respectively. The NL-C26 and NL-B69 genes were also expressed in N. tabacum cv. Samsun nn, which lacks the N gene. Unlike N, NL-C26 and NL-B69, when coexpressed with p50, failed to induce HR on the sites of agroinfiltration in Samsun nn leaves. However, the elicitor-dependent HR in Samsun nn was induced efficiently by chimeric N proteins with the continuous TIR–NBS domains of NL-C26 and NL-B69. On the other hand, the efficiency of HR induction varied significantly among chimeric N proteins with either of the TIR and NBS domains of the NL proteins. In contrast, chimeras carrying the LRR domains of the NL proteins did not induce HR. Thus, the TIR–NBS domains of NL-C26 and NL-B69 could functionally adapt to the LRR domain of N, which may determine the specificity for the elicitor. We speculate that the NL genes are potential HR-inducing resistance genes for undetermined pathogens other than TMV.