Cdk2 activity is essential for the first to second meiosis transition in porcine oocytes

The meiotic progression of Xenopus oocytes has been suggested to depend on the activity of cyclin-dependent kinase 2 (Cdk2). We examined whether Cdk2 is involved in the regulation of mammalian oocyte meiosis by injecting porcine oocytes with anti-Cdk2 antibody. At first, the cross-reactivity of the anti-Cdk2 antibody with Cdc2 kinase was evaluated by immunoprecipitation and immunoblotting experiments using porcine granulosa cell extract, and no cross-reactivity with Cdc2 kinase was observed in the antibody used. In the anti-Cdk2 antibody-injected group, 50.7% of the oocytes were arrested in the second metaphase after 50 h of culture and this rate was significantly lower than those in the non-injected intact oocytes or the oocytes injected with mouse IgG (84.5% and 86.7%, respectively). Most of the other oocytes in the antibody-injected group formed a pronucleus without polar bodies or with only one polar body. The cyclin B1 amount in the antibody-injected and activated oocytes was dramatically decreased compared with that in the intact or mouse IgG-injected oocytes after 50 h of culture. These results suggest that Cdk2 is involved in the meiotic maturation of mammalian oocytes, and that the block of Cdk2 activity results in the failure of cyclin B1 accumulation and second meiosis induction.     

Simulating the BSE epidemic and multiplication factor in dairy herds in Japan

With the objective of evaluating the effectiveness of an administrative guidance on the use of ruminant meat-and-bone meal in ruminant feed, effective from April 1996 to September 2001, we developed a model to simulate the evolution of the BSE epidemic and to estimate the BSE multiplication factor (K) in the Japanese dairy population. The output that provided the best fit to the number of BSE cases both observed and predicted to date suggest that the probability that bovine MBM was fed back to cattle was 14.2-75.2% and 0.129-0.570% during the periods from 1992 to April 1996 and from April 1996 to October 2001, respectively. Given these estimates, the value of K would have peaked in 1995 at 40-48 and then declined to 0.32-0.67 between 1997 and 2001. These results suggest that the administrative guidance was effective in reducing the amount of MBM fed to cattle by a factor of 104-141 and was perhaps enough to drive the epidemic towards extinction.     

Evaluating the contact rate between companion dogs during dog walking and the practices towards potential cases of rabies among dog owners in Japan

This study aimed to examine the habits of dog walking in Japan using an internet survey of insured dog owners. About 96.4% of the respondents (n = 1,151) reported that they would take their dogs for a walk and they most frequently walk their dogs once or twice a day (75.9%) for 30 min to 1 hr (83.1%). The probability of a companion dog having contact with other dogs during dog walking was estimated to be 0.83 (95% confidence interval [CI]: 0.81–0.85), and the associated daily contact rate was estimated using log‐normal distribution with a mean of 2.73 (95% CI: 2.42–3.11) and a standard deviation (SD) of 6.39 (95% CI: 5.18–7.84). Multiple linear regression revealed that the contact rate is mainly influenced by the social behaviour of the owner and to a lesser degree by his/her demographic characteristics including the area of residence, the breed size of dog and the age of the owner. In addition, ten Likert items measured on a 5‐point scale were designed to assess the practices towards potential cases of rabies among dog owners. The respondents (n = 972) achieved a mean score of 2.99 (out of a full score of 4) with a SD of 0.90 in responding to situations related to dog bite incidents and injury from stray cat scratches during dog walking. They achieved a higher score in responding to situations related to sighting a stray or wild animal during dog walking and situations related to non‐specific clinical signs of rabies and bite injuries from stray dogs or wild animals during dog walking with a mean of 3.70 (SD = 0.58) and 3.84 (SD = 0.34), respectively. The level of best practice was also proved to be significantly associated with the demographic characteristics of the dog owner.     

Seroepizootiological survey of Japanese encephalitis virus and Getah virus in regional horse race tracks from 1991 to 1997 in Japan.

A sero-epizootiological survey was conducted for Japanese encephalitis virus (JEV) and Getah virus (GeV) at 10 to 20 regional horse race tracks from 1991 to 1997 in Japan. It was observed that geometrical mean (GM) antibody titer to JEV and GeV was 10 to 50 times higher than others at several race courses (RCs) almost every year. Of them, several race horses showing high antibody titer, which were suggested to be infected with the virus, were also observed in this survey. These data suggested that the viruses have spread among race horses almost every year in Japan, although, fortunately, no horse showing clinical illness due to these viruses was observed. The calendar years and the names of the race courses in which the spread of JEV was suggested were Sonoda and Nakatsu RCs in 1991, Nakatsu RC in 1992, Arao RC in 1993, Nagoya RC in 1994, Kaminoyama, Urawa, Saga and Arao RCs in 1995 and Ooi and Saga RCs in 1997. Spread of JEV was not observed in 1996. The calendar year and name of the race courses at which the spread of GeV was suggested were at Ooi, Sonoda and Nakatsu RCs in 1991, Arao RC in 1992, Nakatsu RC in 1994 and 1995, Funabashi RC in 1996, Saga RC in 1997. It was suggested that surveillance of JEV and GeV should be continued in the future in at least the southern to middle parts of Japan. It has also been suggested that it is necessary to promote the wider use of vaccines to persons related to horse racing.     

The carbohydrate catabolism of <Emphasis Type="Italic">Zostera marina</Emphasis> influenced by lower salinity during the pre-germination stage

The distribution of starch granules in ungerminated seeds of Zostera marina was examined by electron microscopy and histochemistry. Quantitative changes during germination in starch and sugars [glucose/maltosaccharides (MS), sucrose, and fructose] were examined in the seeds using biochemical methods. At natural seawater salinity (30.2 psu), the number of starch granules and the amount of starch markedly decreased, resulting in no increase in the sugar content in the embryos during the early germination stage. A substantial increase in sugars in the shoots occurred after the development of true leaves, probably due to photosynthetic activity in the green true leaves. At natural seawater salinity, the amount of starch did not significantly change during the initial 14 days of germination. In contrast, at lower salinities, starch levels decreased significantly from day 0 to day 6, resulting in higher levels of glucose/MS and fructose in the basal hypocotyls. These results suggest that compared with the salinity of natural seawater, lower salinity may interfere with starch catabolism in seeds, thereby causing the seed coat to break open earlier.     

Expression patterns of the slit subfamily mRNA in canine malignant mammary tumors

Slit, a secreted protein, functions as a chemorepellent factor in axon guidance and neuronal migration and as an inhibitor in leukocyte chemotaxis. In humans, slit2 protein attracts endothelial cells and promotes tube formation in the tumor angiogenic mechanism. In this study, we cloned a part of the canine slit subfamily and examined the expression of slit subfamily mRNAs in 3 normal canine mammary glands and 11 mammary tumor samples by RT-PCR. The cloned part of the slit gene sequences showed high similarity to those of the human, mouse, and rat. The mRNAs were expressed at low levels in the normal mammary gland. The expression levels of slit1 mRNA were low in both the normal and tumor tissues. In contrast, the expression of slit2 mRNA increased in most of the malignant mammary tumors, and an increase in slit3 mRNA expression was observed in 2 of the malignant mixed tumors. These results suggest that the expression of slit2 plays an important role in tumor angiogenesis in canine mammary gland tumors and that slit2 can be a putative marker for malignancy diagnosis of these tumors.     

Adjusted incidence risks of BSE in risk subpopulations of cattle in Japan

Immediately after the detection of the first case of BSE in 2001, the Japanese government introduced active surveillance targeting fallen-stock and all cattle slaughtered for human consumption. By the end of 2004, four million animals were tested with rapid tests under the passive and active surveillance. As a result 13 additional cases were detected. I focused on the 1996 birth cohort, in which nine cases of BSE were detected during 2001–2004, and estimated the prevalence of BSE infection of that birth cohort using maximum-likelihood methods. Using the estimated prevalence of infection as an input variable, I calculated the adjusted incidence risk of BSE by different ages and risk subpopulations (clinical-suspects, fallen-stock, sick-slaughter and healthy-slaughter animals). The adjusted incidence risk of BSE in sick-slaughter animals (animals showing clinical signs not compatible with BSE when slaughtered for human consumption) was 18.7 and 4.5–78.4 times higher than the incidence risk in fallen-stock and healthy-slaughter animals, respectively.