Canine epididymal sperm was collected from the cauda epididymis using 2 different methods (flushing and mincing) to compare the qualities (the percentage of progressively motile, viable, morphologically abnormal, immature and intact acrosomes) before and after freezing and thawing. No significant difference was noted in the quality of the cauda epididymal sperm immediately after collection and after freezing-thawing between the collection methods, although the mean levels of sperm quality with the flushing method were slightly better than that of the mincing method. The flushing method is simple and free of blood contamination, although the vas deferens was too small to be perfused in only 1 dog, and our results suggest that the flushing method is preferable to the mincing method for collecting sperm from the canine cauda epididymis.
It is found that a heartbeat period fluctuation usually has a power spectral density which is inversely proportional to frequency, to which a spike is added at a breathing frequency.
Human infections with the H5 highly pathogenic avian influenza virus (HPAIV) sporadically threatens public health. The susceptibility of HPAIVs to baloxavir acid (BXA), which is a new class of inhibitor for the influenza virus cap-dependent endonuclease, has been confirmed in vitro, but has not yet been characterized fully. Here, the efficacy of BXA against HPAIVs, including recent H5N8 variants in vitro was assessed. The antiviral efficacy of baloxavir marboxil (BXM) in H5N1 virus-infected mice was also investigated. BXA exhibited similar in vitro activities against H5N1, H5N6, and H5N8 variants tested to those of seasonal and other zoonotic strains. BXM monotherapy in mice infected with the H5N1 HPAIV clinical isolate; A/Hong Kong/483/1997 (H5N1) strain, also caused a significant reduction in viral titers in the lungs, brains, and kidneys, followed by prevention of acute lung inflammation and improvement of mortality compared with oseltamivir phosphate (OSP). Furthermore, combination treatments with BXM and OSP, using a 48-hour delayed treatment model showed a more potent effect on viral replication in organs, accompanied by improved survival compared to BXM or OSP monotherapy. From each test, no resistant virus (e.g., I38T in the PA) emerged in any BXM-treated mouse. These results therefore support the conclusion that BXM has potent antiviral efficacy against H5 HPAIV infections.
The molecular mechanism of muscle degeneration in a lethal muscle disorder Duchene muscular dystrophy (DMD) has not been fully elucidated. The dystrophic dog, a model of DMD, shows a high mortality rate with a marked increase in serum creatine kinase (CK) levels in the neonatal period. By measuring serum CK levels in cord and venous blood, we found initial pulmonary respiration resulted in massive diaphragm damage in the neonates and thereby lead to the high serum CK levels. Furthermore, molecular biological techniques revealed that osteopontin was prominently upregulated in the dystrophic diaphragm prior to the respiration, and that immediate-early genes (c-fos and egr-1) and inflammation/immune response genes (IL-6, IL-8, COX-2, and selectin E) were distinctly overexpressed after the damage by the respiration. Hence, we segregated dystrophic phases at the molecular level before and after mechanical damage. These molecules could be biomarkers of muscle damage and potential targets in pharmaceutical therapies.
The importance of knee meniscus function is now recognized, and the treatment of meniscus injury has been changing from resection to repair. However, depending on the type of injury, meniscectomy sometimes cannot be avoided. In such case, it is important to anticipate the future problem of degenerative change or osteoarthrosis in knee joint. In future, auto-graph using regenerative meniscus will be developed by tissue engineering. However, even if it will be possible, considering the healing period, the young athlete with severe meniscus injury may select meniscectomy in order to return to sports life as early as possible avoiding the long immobilization or declining sports skill. In consideration of the prognosis and circumstances in such patients, we need a artificial meniscus for salvage. To assess further the use of polyvinyl alcohol-hydrogel (PVA-H) artificial meniscus, we performed some mechanical tests about PVA-H and animal experiment. In mechanical tests, we found that a high water content PVA-H showed viscoelastic behaviour similar to that of human meniscus. Moreover, the frictional coefficient of PVA-H against natural articular cartilage was also effective. In the animal experiment using rabbits, the lateral meniscus was replaced with an artificial meniscus in one knee side and lateral meniscectomy was performed in another knee side of each rabbit. We have already reported the results up to 1 year after operation, present study investigated the results in postoperative 1.5 years. In the results, the articular cartilage state of knee joint implanted PVA-H meniscus was good even after 1.5 years, while OA change progressed in meniscectomy knee joint. In addition, neither wear nor breakage of PVA-H was observed. These results proved that an artificial meniscus using a high water content PVA-H can compensate for meniscal function and might be clinically applicable.
Monoclonal antibodies (MAb) against A cantonensis were produced through fusion of immunised spleen cells from BALB/c mice with NS-1 myeloma cells at a ratio of 10:1. The successful fusion rate on the 3rd day of fusion was 90.1%. Ten MAb were characterised, six of which were IgG1 and the remaining four were IgG2a, IgG2b, IgM and IgA respectively. Among 6 IgG1 MAb, four were A. cantonensis-specific, of which three reacted to adult worm antigen only and one reacted to both adult worm and juvenile worm antigens. Two other IgG1 MAb showed cross-reaction with other helminthic antigens of Toxocara canis. Ascaris suum. Paragonimus westermani, Dirofilaria immitis, Anisakis Spp, Gnatostoma Spinigerum and Clonorchis sinensis.
