Quantitative risk assessment of the introduction of rabies into Japan through the importation of dogs and cats worldwide.

Japan has been free from rabies since 1958. A strict import regimen has been adopted since 2004 consisting of identification of an animal with microchip, two-time rabies vaccination, neutralizing antibody titration test and a waiting period of 180 days. The present study aims to quantitatively assess the risk of rabies introduction into Japan through the international importation of dogs and cats and hence provide evidence-based recommendations to strengthen the current rabies prevention system. A stochastic scenario tree model was developed and simulations were run using @RISK. The probability of infection in a single dog or cat imported into Japan is estimated to be 2·16 × 10-9 [90% prediction interval (PI) 6·65 × 10-11-6·48 × 10-9]. The number of years until the introduction of a rabies case is estimated to be 49 444 (90% PI 19 170-94 641) years. The current import regimen is effective in maintaining the very low risk of rabies introduction into Japan and responding to future changes including increases in import level and rabies prevalence in the world. However, non-compliance or smuggling activities could substantially increase the risk of rabies introduction. Therefore, policy amendment which could promote compliance is highly recommended. Scenario analysis demonstrated that the waiting period could be reduced to 90 days and the requirement for vaccination could be reduced to a single vaccination, but serological testing should not be stopped.

A current life table and causes of death for insured dogs in Japan.

The life expectancies and causes of death were evaluated in 299,555 dogs insured in Japan between 1 April 2010 and 31 March 2011, of which 4169 dogs died during this period. The overall life expectancy of dogs was 13.7 years. The probability of death was high in the first year of life, lowest in the second and third years, and increased exponentially after 3 years of age. The life expectancy was 13.8 years in the <5 kg body weight group, 14.2 years in the 5-10 kg body weight group, 13.6 years in the 10-20 kg body weight group, 12.5 years in the 20-40 kg body weight group and 10.6 years in the ≥40 kg body weight group. As body weight increases, life expectancy tended to decrease except in the <5 kg body weight group. The probability of death increased as dogs got older for most potential causes of death. Neoplasia resulted in the highest probability of death, especially in the large and giant breed groups. Cardiovascular system disorders were the second major cause of death and the toy group had a probability of death significantly higher than the other breed groups at age 12+.

Breed, gender and age pattern of diagnosis for veterinary care in insured dogs in Japan during fiscal year 2010.

We calculated the annual prevalence of diseases of 18 diagnostic categories in the insured dog population in Japan, using data from 299,555 dogs insured between April 2010 and March 2011. The prevalence was highest for dermatological disorders (22.6% for females and 23.3% for males), followed by otic diseases (16.4% for females and 17.2% for males) and digestive system disorders (15.7% for females and 16.4% for males). The prevalence of cardiovascular, urinary, neoplasia and endocrine disorders, increased with age; infectious diseases and injuries showed a high prevalence at young ages, and the prevalence of musculoskeletal and respiratory disorders showed a bimodal peak at young and old ages. A large variation in prevalence was observed between breeds for dermatological, otic, digestive, ophthalmological and cardiovascular disorders.

Sales of veterinary antimicrobial agents for therapeutic use in food-producing animal species in Japan between 2005 and 2010.

The use of veterinary antimicrobial agents in animals can result in the emergence and selection of resistant bacteria in food-producing animals. This study elucidated the use of veterinary antimicrobial agents in Japan in terms of milligrams of active ingredient sold per kilogram of biomass between 2005 and 2010. Data on sales of antimicrobial agents and on the biomass of the target animal species were compiled from statistics published bythe Japanese Ministry of Agriculture, Forestry and Fisheries. The quantities of antimicrobials used varied between animal species: the highest usage was observed in pigs (392 to 423 mg/ kg), followed by beef cattle (45 to 67 mg/kg), broiler chickens (44 to 63 mg/kg) and dairy cattle (33 to 49 mg/kg). For the animal species combined, usage of third- and fourth-generation cefalosporins, fluoroquinolones and macrolides ranged from 0.10 to 0.14 mg/kg biomass, 1.1 to 1.3 mg/kg biomass and 7.8 to 10.6 mg/kg biomass, respectively.

The influence of reduced glutathione in fertilization medium on the fertility of in vitro-matured C57BL/6 mouse oocytes.

It is well known that IVM oocytes show a decreased potential for fertility and development compared with in vivo-matured oocytes. In this study, we added reduced glutathione (GSH) to the fertilization medium during IVF to investigate its effect on the fertility and early embryo development of IVM oocytes. The fertilization rate for IVM oocytes and fresh sperm increased with the addition of GSH (0, 1.0, and 2.0 mM: 51%, 76%, and 70%). Moreover, the addition of GSH to the fertilization medium also improved the developmental potential compared with the control sample (0 mM). In addition, we performed IVF using IVM oocytes and frozen/thawed sperm that had been cryopreserved in a mouse bank. Results indicated a marked increase in the fertilization rate when 1.0 mM GSH was added to the fertilization medium compared with when no GSM was used (0.0 mM GSH: 2% (3/195); 1.0 mM GSH: 33% (156/468)). Furthermore, the fertilization rate improved dramatically via zona drilling using laser equipment (52%: 267/516), whereas normal offspring were obtainsed after transferring embryos created via IVF using IVM oocytes and frozen/thawed sperm. This is the first report in which offspring have been obtained via IVF using IVM oocytes and frozen/thawed sperm.

Growth and development of rabbit oocytes in vitro: effect of fetal bovine serum concentration on culture medium.

The objective was to develop a culture system that produced blastocyst stage embryos from rabbit oocytes grown in vitro. Two experiments were performed. First, various concentrations of fetal bovine serum (FBS, 0, 0.05, 0.5 and 5%) were used in the culture medium for in vitro growth (IVG) of oocytes recovered from follicles 200 to 299 µm in diameter. Intracytoplasmic sperm injection (ICSI) was performed on mature oocytes obtained after IVG for 8 days and in vitro maturation for 14 to 16 h. Rates of survival and pronuclear formation after ICSI were higher for oocytes grown in a medium with 0.05% FBS compared to oocytes grown in a medium lacking FBS (97.6 vs. 76.9%, 97.5 vs. 70%, P < 0.1). The rate of development to the blastocyst stage was also higher in the medium containing 0.05% FBS than in the medium lacking FBS (9.5 vs. 17.9%, P < 0.05). Next, using oocytes recovered from follicles 200 to 399 µm in diameter which were cultured in 0.05% FBS, oxygen consumption and the number of cells were analyzed. Blastocysts from oocytes grown in vitro with 0.05% FBS had reduced oxygen consumption and number of cells compared with those from ovulated oocytes (21.66 ± 4.54 × 10(14) vs. 50.19 ± 4.61 × 10(14) mol/sec, 244 ± 25 vs. 398 ± 24, P < 0.05). Rabbit oocytes grown in vitro with 0.05% FBS achieved pregnancy, but pregnancies were not maintained to term. In conclusion, the addition of 0.05% FBS to the culture medium for IVG improved developmental competence of rabbit oocytes grown in vitro.

Edaravone, a known free radical scavenger, enhances X-ray-induced apoptosis at low concentrations.

Edaravone has been reported to have a radioprotective effect at high concentrations. We now report that a lower dose of edaravone enhanced X-ray-induced apoptosis of some cell lines harboring p53 wild-type status, such as MOLT-4, Nalm-6, and HepG2. The knock-down of p53 using siRNA in MOLT-4 cells abolished the radiosensitizing effect of edaravone. Enhanced phosphorylations of p53 at Ser 15 and Ser 20 and up-regulation of PUMA, a p53 target protein, were observed after X-irradiation in the presence of edaravone. We conclude that the low dose of edaravone sensitized cells to X-irradiation by promoting the p53-dependent apoptotic signaling pathway.

[Tricuspid annuloplasty, pulmonary valve replacement, ventricular septal defect (VSD) patch closure, and right-sided maze procedure 23-years after corrective repair of tetralogy of Fallot].

Postoperative pulmonary valve regurgitation, stenosis of the right ventricular outflow tract, conduit failure, ventricular septal patch leak, secondary tricuspid valve regurgitation, and various arrhythmias are the major complications that develop after surgical repair of tetralogy of Fallot in adults. A 27-year-old male with pulmonary regurgitation, tricuspid regurgitation, residual ventricular septal defect (VSD), low left ventricular function, and chronic atrial fibrillation underwent tricuspid annuloplasty, pulmonary valve replacement with a stentless aortic valve, VSD patch closure, and right-sided maze procedure, and the postoperative course was uneventful. The cardiothoracic ratio decreased, sinus rhythm was restored, and the patient's complaints were relieved. Reoperation at the optimal time after corrective repair of tetralogy of Fallot in adults may improve the outcome.

Negative regulation of MEKK1/2 signaling by serine-threonine kinase 38 (STK38).

Mitogen-activated protein kinases (MAPKs) are activated through the kinase cascades of MAPK, MAPK kinase (MAPKK) and MAPKK kinase (MAPKKK). MAPKKKs phosphorylate and activate their downstream MAPKKs, which in turn phosphorylate and activate their downstream MAPKs. MAPKKK proteins relay upstream signals through the MAPK cascades to induce cellular responses. However, the molecular mechanisms by which given MAPKKKs are regulated remain largely unknown. Here, we found that serine-threonine protein kinase 38, STK38, physically interacts with the MAPKKKs MEKK1 and MEKK2 (MEKK1/2). The carboxy terminus, including the catalytic domain, but not the amino terminus of MEKK1/2 was necessary for the interaction with STK38. STK38 inhibited MEKK1/2 activation without preventing MEKK1/2 binding to its substrate, SEK1. Importantly, STK38 suppressed the autophosphorylation of MEKK2 without interfering with MEKK2 dimer formation, and converted MEKK2 from its phosphorylated to its nonphosphorylated form. The negative regulation of MEKK1/2 was not due to its phosphorylation by STK38. On the other hand, stk38 short hairpin RNA enhanced sorbitol-induced activation of MEKK2 and phosphorylation of the downstream MAPKKs, MKK3/6. Taken together, our results indicate that STK38 negatively regulates the activation of MEKK1/2 by direct interaction with the catalytic domain of MEKK1/2, suggesting a novel mechanism of MEKK1/2 regulation.

Three major dietary patterns are all independently related to the risk of obesity among 3760 Japanese women aged 18-20 years.

OBJECTIVE: To examine associations between dietary patterns and obesity. DESIGN: Cross-sectional study. SUBJECTS: A total of 3760 Japanese female dietetic course students aged 18-20 years from 53 institutions in Japan. MEASUREMENTS: Diet was assessed over a 1-month period with a validated self-administered diet history questionnaire with 148 food items, from which 30 food groups were created and entered into a factor analysis. Body mass index (BMI) was calculated from self-reported body height and weight. RESULTS: Mean BMI (+/-s.d.) was 20.9+/-2.8 kg m(-2). Four dietary patterns were identified. After adjustment for several confounding factors and total energy intake, the 'Healthy' pattern, characterized by high intakes of vegetables, mushrooms, seaweeds, potatoes, fish and shellfish, soy products, processed fish, fruit and salted vegetables, was significantly associated with a lower risk of BMI> or =25 (odds ratio of the highest quintile vs lowest, 0.57; 95% confidence interval: 0.37-0.87; P for trend <0.05). In contrast, the 'Japanese traditional' pattern, characterized by high intakes of rice, miso soup and soy products, and the 'Western' pattern, characterized by high intakes of meats, fats and oils, seasonings, processed meats and eggs, were both significantly associated with an increased risk of BMI> or =25 (OR: 1.77; 95% CI: 1.17-2.67; P for trend <0.01 and OR: 1.56; 95% CI: 1.01-2.40; P for trend=0.04, respectively). CONCLUSION: Three major dietary patterns, Healthy, Japanese traditional and Western, were all independently and significantly related to the risk of obesity even among a relatively lean young Japanese female population.

Dietary fiber intake, dietary glycemic index and load, and body mass index: a cross-sectional study of 3931 Japanese women aged 18-20 years.

OBJECTIVE: Few observational studies have investigated dietary fiber intake and dietary glycemic index (GI) and glycemic load (GL) simultaneously in relation to obesity, particularly in non-Western populations. We examined the associations between dietary fiber intake and dietary GI and GL, and body mass index (BMI) in young Japanese women. DESIGN: Cross-sectional study. SUBJECTS: A total of 3931 female Japanese dietetic students aged 18-20 years from 53 institutions in Japan. METHODS: Dietary fiber intake and dietary GI and GL (GI for glucose=100) were assessed by a validated, self-administered, diet history questionnaire. BMI was calculated from self-reported body weight and height. RESULTS: Mean values of BMI, dietary fiber intake, dietary GI and dietary GL were 21.0 kg/m(2), 6.5 g/4186 kJ, 65.1 and 82.1/4186 kJ, respectively. White rice (GI=77) was the major contributor to dietary GI and GL (45.8%). After controlling for potential dietary and nondietary confounding factors, dietary fiber intake was negatively correlated with BMI (adjusted mean=21.1 kg/m(2) in the lowest and 20.7 kg/m(2) in the highest quintiles; P for trend=0.0007). Conversely, dietary GI and GL were independently positively correlated with BMI (20.8 and 21.2 kg/m(2); P for trend=0.03, and 20.5 and 21.5 kg/m(2); P for trend=0.0005, respectively). CONCLUSIONS: Dietary fiber intake showed an independent negative association with BMI, and dietary GI and GL showed an independent positive association with BMI among relatively lean young Japanese women.

Association between dietary fiber, water and magnesium intake and functional constipation among young Japanese women.

OBJECTIVE: Most research on constipation has focused on dietary fiber intake. Here, we examined the intake of water and magnesium, nutrients possibly associated with constipation, as well as that of dietary fiber in relation to constipation. DESIGN: Cross-sectional study. SUBJECTS: A total of 3835 female Japanese dietetic students aged 18-20 years from 53 institutions in Japan. METHODS: Dietary intake was estimated with a validated, self-administered diet history questionnaire. Functional constipation was defined using the Rome I criteria. RESULTS: The prevalence of functional constipation was 26.2%. Neither dietary fiber intake (mean=6.4 g/4186 kJ) nor intakes of total water and water from fluids were associated with constipation. Conversely, low intake of water from foods was associated with an increasing prevalence of constipation. In comparison with women in the first (lowest) quintile, the multivariate adjusted odds ratio (OR) (95% confidence interval (CI)) for women in the second, third, fourth, and fifth quintiles were 0.72 (0.57, 0.90), 0.78 (0.62, 0.98), 0.71 (0.56, 0.89), and 0.77 (0.61, 0.97), respectively (P for trend=0.04). Additionally, low magnesium intake was associated with increasing prevalence of constipation. Compared with women in the first quintile, the multivariate adjusted OR (95% CI) for women in the second, third, fourth and fifth quintiles were 0.70 (0.56, 0.88), 0.75 (0.60, 0.95), 0.73 (0.58, 0.92) and 0.79 (0.63, 0.996), respectively (P for trend=0.09). CONCLUSIONS: Low intakes of water from foods and magnesium are independently associated with an increasing prevalence of functional constipation among a population whose dietary fiber intake is relatively low.

Preservation of ovarian function by ovarian shielding when undergoing total body irradiation for hematopoietic stem cell transplantation: a report of two successful cases.

The purpose of this study was to evaluate the possibility of preserving ovarian function by ovarian shielding to reduce the irradiation dose in total body irradiation (TBI). The subjects in the study were females aged less than 40 years, who were undergoing allogeneic hematopoietic stem cell transplantation using a TBI-based regimen and who desired to have children after transplantation. For ovarian shielding, abdominal computed tomography (CT) and skin marking were performed in both the supine and prone positions, prior to the TBI. A pair of columnar blocks was placed just above the patient's body. Thus far three patients have been treated. The serum estradiol level decreased to an undetectable level (<8.5 pg/ml) after transplantation and the follicle-stimulating hormone (FSH) level increased above 90 mIU/ml in all patients and they became amenorrheic. However, regular menstruation recovered in patients no. 1 and 2 about 800 and 370 days after transplantation, respectively, with a decrease in the serum FSH level. Menstruation did not recover in patient no. 3, and serum estradiol was transiently detected above 20 pg/ml. The preservation of ovarian function was made possible by ovarian shielding. However, a longer follow-up is needed to know if normal pregnancy and delivery can occur.

Sodium orthovanadate suppresses DNA damage-induced caspase activation and apoptosis by inactivating p53.

We previously reported that p42/SETbeta is a substrate for caspase-7 in irradiated MOLT-4 cells, and that treating the cells with sodium orthovanadate (vanadate) inhibits p42/SETbeta's caspase-mediated cleavage. Here, we initially found that the inhibitory effect of vanadate was due to the suppression of caspase activation but not of caspase activity. Further investigations revealed that vanadate suppressed upstream of apoptotic events, such as the loss of mitochondrial membrane potential, the conformational change of Bax, and p53 transactivation, although the accumulation, total phosphorylation, and phosphorylation of six individual sites of p53 were not affected. Importantly, vanadate suppressed p53-dependent apoptosis, but not p53-independent apoptosis. Finally, gel-shift and chromatin immunoprecipitation assays conclusively demonstrated that vanadate inhibits the DNA-binding activity of p53. Vanadate is conventionally used as an inhibitor of protein tyrosine phosphatases (PTPs); however, we recommend that the influence of vanadate not only on PTPs but also on p53 be considered before using it.

Impaired ambulatory venous function in lymphedema assessed by near-infrared spectroscopy.

AIM: The aim of this study was to investigate venous function in patients with leg lymphedema during exercise using near-infrared spectroscopy (NIRS), compared with that of patients with chronic venous insufficiency (CVI). METHODS: Forty-three legs of 33 patients (5 males, 28 females; mean age: 53 years) with leg lymphedema without varicose veins or deep vein thrombosis underwent a treadmill walking test with simultaneous NIRS. For comparison, 136 legs of 91 patients (35 males, 56 females; mean age: 56 years) with varicose veins as a CVI group and 45 legs of 38 healthy volunteers (23 males, 15 females; mean age: 50 years) were also evaluated in the same method. Deoxygenated hemoglobin (HHb) was continuously measured during exercise, and the ambulatory venous retention index (AVRI) of each leg was obtained from the serial changes in HHb. RESULTS: The mean AVRI of the lymphedema group was significantly higher than that of healthy legs and significantly lower than of legs with moderate or severe CVI. Furthermore, it was similar to that in the mild CVI group. CONCLUSIONS: Venous function is impaired in exercising legs with lymphedema, and corresponds to that in legs with mild venous insufficiency. The treatment of lymphedema should take CVI into consideration.

Isolation of equine herpesvirus-1 lacking glycoprotein C from a dead neonatal foal in Japan.

We isolated a variant equine herpesvirus-1 (EHV-1), strain 5089, from the lung of a dead neonatal foal in Japan and characterized the biological nature of the virus. The virus spread in cultured cells mainly by cell-to-cell infection, unlike wild-type EHV-1, which spreads efficiently as a cell-free virus. The virus titer in cultured supernatant and the intracellular virus titer were low compared to those of wild-type EHV-1. Heparin treatment of the virus had no effect on viral infectivity in cell culture. Glycoprotein C (gC) was not detected by Western blotting and fluorescent antibody tests in 5089 virions and 5089-infected cells, respectively. RT-PCR analysis revealed that the expression level of 5089 gC mRNA was reduced considerably compared to that of wild-type EHV-1. Sequencing analysis of the 5089 gC coding region showed a point mutation in the promoter region of the gC open reading frame. However, the mutation did not affect the promoter activity. These results suggested that the lack of gC in 5089 virions might be one of the reasons for spread of the virus by cell-to-cell infection and that gC mRNA expression might not be activated efficiently due to factors other than the mutation in the gC promoter region.

A study of SS size distribution during runoff and fractionation of phosphates depending on soil size in agricultural watershed.

Characterization of the differences and algal-available fractions of P in soils, suspended solids, and bottom sediments have been the main topics of research during the past decade. However, the size distribution and properties of particulate matter in runoff have not been much studied in Japan. Here we study particle size distribution during runoff and the chemical characteristics of P in each soil size fraction and relate them to land use. The temporal variation of particulate sizes during rain events is different in each watershed. Most particles have the size in the range of 10-100 microm. Also, the percentage of BAP in TP as well as percentage of PCOD in SS also varies temporally and spatially during runoff. To investigate how soil particles characteristics depend on land use, soil samples from two watersheds are examined. For particle size distribution and specific gravity, no significant difference among watersheds is found. However, C, N, and P content are indirectly proportional to the particle size, which means smaller particle size results in larger. H2O-extracted P, NH4Cl-extracted P, NAI-P, Apatite-P, Organic-P, and TP contents in each soil particle sample vary depending on particle size, land use, and watershed.

Dissolved organic matter from agricultural fields in the irrigation period.

The aim of this study was to quantify and characterize the dissolved organic carbon (DOC) of paddy fields and crop fields in Tottori, Japan. Dissolved organic carbon (DOC) and ultraviolet (UV) absorbance was measured for the filtrated water of each samples. DOC concentration and SUVA (specific UV absorbance) of biodegradation analysis samples were determined around 50 days after the incubation. In the Fukui paddy fields, DOC concentration varied seasonally from 1.1 to 10.1 mg.Cl(-1), showing higher concentration in heavy runoff of non-agriculture period in April. However, DOC concentration variation did not always correspond to rainfall. The Obadake paddy fields also showed a similar pattern with Fukui paddy fields. The daily DOC discharge per area in Fukui (up), Fukui (down), Obadake (south), Obadake (north) paddy fields influent from paddy fields were 0.02, 0.0161, 0.0135 and 0.0027 kg.a(-1).day(-1), respectively. These differences resulted from differences in agricultural types and customs of farmers according to paddy fields and fields. Also, SUVA (an indirect means to evaluate humic substances (hydrophobic fraction)) of the studied influent waters from paddy fields were generally lower than the influent waters from crop fields. The non-biodegradable DOC accounted for 50.2-98%, 46.8-85.5% of the total DOC in the paddy fields and crop fields.