Geriatric nutritional risk index as the prognostic factor in older patients with fragility hip fractures.

This study investigated the long-term survival and incidence of secondary fractures after fragility hip fractures. The 5-year survival rate was 62%, and the mortality risk was seen in patients with GNRI < 92. The 5-year incidence of secondary fracture was 22%, which was significantly higher in patients with a BMI < 20. BACKGROUND: Malnutrition negatively influences the postoperative survival of patients with fragility hip fractures (FHFs); however, little is known about their association over the long term. OBJECTIVE: This study evaluated the ability of the geriatric nutritional risk index (GNRI) as a risk factor for long-term mortality after FHFs. METHODS: This study included 623 Japanese patients with FHFs over the age of 60 years. We prospectively collected data on admission and during hospitalization and assessed the patients' conditions after discharge through a questionnaire. We examined the long-term mortality and the incidence of secondary FHFs and assessed the prognostic factors. RESULTS: The mean observation period was 4.0 years (range 0-7 years). The average age at the time of admission was 82 years (range 60-101 years). The overall survival after FHFs (1 year, 91%; 5 years, 62%) and the incidence of secondary FHFs were high (1 year, 4%; 5 years, 22%). The multivariate Cox proportional hazard analysis revealed the risk factors for mortality as older age (hazard ratio [HR] 1.04), male sex (HR 1.96), lower GNRI score (HR 0.96), comorbidities (malignancy, HR 2.51; ischemic heart disease, HR 2.24; revised Hasegawa dementia scale ≤ 20, HR 1.64), no use of active vitamin D3 on admission (HR 0.46), and a lower Barthel index (BI) (on admission, HR 1.00; at discharge, HR 0.99). The GNRI scores were divided into four risk categories: major risk (GNRI, < 82), moderate risk (82-91), low risk (92-98), and no risk (> 98). Patients at major and moderate risks of GNRI had a significantly lower overall survival rate (p < 0.001). Lower body mass index (BMI) was also identified as a prognostic factor for secondary FHFs (HR 0.88 [p = 0.004]). CONCLUSIONS: We showed that older age, male sex, a lower GNRI score, comorbidities, and a lower BI are risk factors for mortality following FHFs. GNRI is a novel and simple predictor of long-term survival after FHFs.

An economic assessment of foot and mouth disease in Japan.

A large-scale foot and mouth disease (FMD) epidemic in Japan in 2010 caused severe economic losses for livestock and related industries. In this paper, the authors develop a clear and usable framework to estimate the economic impact of this FMD outbreak. An economic analysis is then conducted by combining this framework with an epidemiological model. The framework estimates the direct and indirect costs to livestock and related industries by applying an input-output model, as well as by addressing expenditure on disease control. The direct cost to the livestock industry was estimated at 51.2 billion Japanese yen (JPY), engendering an indirect cost to related industries of JPY 25.5 billion. The expenditure for disease control activities was estimated at JPY 8.2 billion. The total impact of the 2010 FMD epidemic was estimated at almost JPY 85 billion. Within the economic analysis, the authors evaluate several control measure scenarios: a baseline scenario, which assumes that the rapid disease spread observed in the early phase of the 2010 FMD epidemic would continue; prompt culling within 24 hours; early detection of the first case; and emergency vaccination within a radius of 10 km around the affected farms in either seven or 28 days. Prompt culling and early detection were superior from an economic point of view, reducing the total economic impact to 30% and 2% of that in the baseline scenario, respectively. Compared with these scenarios, vaccination was less cost effective. However, vaccination suppressed the speed of disease spread and shortened the duration of the epidemic, suggesting its potential effectiveness in curbing rapid disease spread in a densely populated area.

Une épizootie de fièvre aphteuse de grande envergure survenue au Japon en 2010 a entraîné de graves pertes économiques pour la filière de l'élevage et les secteurs connexes. Les auteurs proposent un cadre d'évaluation clair pour estimer l'impact économique de ce foyer de fièvre aphteuse. Ils effectuent ensuite une analyse économique dans laquelle ce cadre d'évaluation est relié à un modèle épidémiologique. Les coûts directs et indirects subis par la filière de l'élevage et par les secteurs connexes sont évalués dans ce cadre en appliquant un modèle entrées­sorties (input­output) et en prenant en compte les dépenses liées à la lutte contre la maladie. Le coût direct de la maladie pour la filière de l'élevage a été estimé à 51,2 milliards de yens japonais, auxquels s'ajoute le coût indirect pour les secteurs connexes estimé à 25,5 milliards de yens japonais. Les dépenses induites par la lutte contre la maladie s'élèvent à 8,2 milliards de yens japonais. L'impact total de l'épizootie de fièvre aphteuse de 2010 est estimé à près de 85 milliards de yens japonais. Dans leur analyse économique, les auteurs ont évalué plusieurs scénarios relatifs aux mesures de lutte appliquées : un scénario de base, qui suppose une propagation de la maladie au même rythme que durant la phase initiale de l'épidémie de 2010 ; l'abattage rapide des animaux en 24 heures ; la détection précoce du premier cas ; l'application de la vaccination d'urgence dans un périmètre de 10 km autour des exploitations affectées et dans un délai de sept ou de 28 jours. Par rapport au scénario de base, l'abattage rapide et la détection précoce sont les meilleurs scénarios du point de vue économique, réduisant l'impact économique total respectivement à 30 % et 2 % de celui du scénario de base. Comparativement à ces scénarios, la vaccination est la solution la moins efficiente économiquement. Néanmoins, la vaccination a freiné la propagation de la maladie et limité la durée de l'épidémie, ce qui laisse penser qu'elle permettrait d'infléchir la courbe de propagation de la maladie dans les zones à forte densité de bétail.

En 2010 se produjo en el Japón una epidemia de fiebre aftosa de grandes proporciones, que causó graves pérdidas económicas a la industria ganadera y demás sectores conexos. Los autores describen un método claro para estimar el impacto económico de dicho brote. Acto seguido, llevan a cabo un análisis económico combinando este método con un modelo epidemiológico. El método permite estimar los costos directos e indirectos que soportan la industria ganadera y demás sectores conexos aplicando un modelo de «insumoproducto ¼ (input­output) y teniendo también en cuenta los gastos ligados al control de la enfermedad. Según los cálculos, el costo directo para la industria ganadera fue de 51 200 millones de yenes japoneses y el costo indirecto para los sectores conexos fue de 25 500 millones de yenes. Se estimó que las actividades de lucha contra la enfermedad supusieron gastos por valor de 8 200 millones. En total, pues, según estas estimaciones, el impacto económico de la epidemia de fiebre aftosa de 2010 se cifra en casi 85 000 millones de yenes. Como parte del análisis económico, los autores evalúan varias hipótesis relativas a las medidas de control: una hipótesis básica, en la cual prosigue sin trabas la veloz propagación de la enfermedad observada en la primera fase de la epidemia de 2010; la rápida implantación, en un plazo de 24 horas, de medidas de sacrificio sanitario; la pronta detección del primer caso; y la vacunación de emergencia dentro de un radio de 10 km alrededor de las explotaciones afectadas, en un plazo de siete días o en un plazo de 28 días. Desde un punto de vista económico, el rápido sacrificio sanitario y la pronta detección ofrecían mejores resultados, pues reducían el impacto económico total hasta un 30% y un 2%, respectivamente, del registrado en la hipótesis básica. En comparación con esas dos hipótesis, la vacunación ofrecía menos eficacia en relación con el costo, aunque frenaba la rápida propagación de la enfermedad y abreviaba la epidemia, de donde se infiere que puede resultar eficaz para contener la rápida dispersión de la enfermedad en áreas densamente pobladas.

Bone morphogenetic protein 7 induces cementogenic differentiation of human periodontal ligament-derived mesenchymal stem cells.

Bone morphogenetic protein 7 (BMP-7) is a multifunctional differentiation factor that belongs to the transforming growth factor superfamily. BMP-7 induces gene expression of protein tyrosine phosphatase-like, member A/cementum attachment protein (PTPLA/CAP) and cementum protein 1 (CEMP1), both of which are markers of cementoblasts and cementocytes. In the previous study, we reported that BMP-7 treatment enhanced PTPLA/CAP and CEMP1 expression in both normal and immortal human periodontal ligament (PDL) cells. To elucidate the molecular mechanisms of the gene expression of these molecules, in this study, we identified a functional transcription activator binding region in the promoter region of PTPLA/CAP and CEMP1 that is responsive to BMP signals. Here, we report that some short motifs termed GC-rich Smad-binding elements (GC-SBEs) that are located in the human PTPLA/CAP promoter and CEMP1 promoter are BMP-7 responsive as analyzed with luciferase promoter assays. On the other hand, we found that transcription of Sp7/Osterix and PTPLA/CAP was up-regulated after 1 week of BMP-7 treatment on purified normal human PDL cells as a result of gene expression microarray analysis. Furthermore, transcription of Sp7/Osterix, runt-related transcription factor 2 (RUNX2), and alkaline phosphatase (ALP) was up-regulated after 2 weeks of BMP-7 treatment, whereas gene expression of osteo/odontogenic markers such as integrin-binding sialoprotein (IBSP), collagen, type I, alpha 1 (COL1A1), dentin matrix acidic phosphoprotein 1 (DMP1), and dentin sialophosphoprotein (DSPP) was not up-regulated in purified normal or immortal human PDL cells as a result of qRT-PCR. The results suggest that BMP-7 mediates cementogenesis via GC-SBEs in human PDL cells and that its molecular mechanism is different from that for osteo/odontogenesis.

Injuries to staff engaged in foot-and-mouth disease eradication in Japan.

BACKGROUND: In April 2010, a large-scale outbreak of foot-and-mouth disease, a highly infectious disease affecting cloven-hoofed animals, was reported in Miyazaki prefecture in Japan. Many staff were dispatched to the epidemic area to undertake containment measures. Various injuries were reported among the dispatched staff. AIMS: To study the characteristics of injuries that occurred during containment measures and to identify the characteristics of those injured. METHODS: We analysed records of injuries that occurred among staff dispatched from national organizations and prefectures other than Miyazaki prefecture. Based on these records, a qualitative analysis was conducted for veterinarians and non-veterinarian support staff as each group played a different role during the outbreak. RESULTS: Forty-seven veterinarians and 183 support staff were injured. The injury risk was significantly higher for support staff (13%) than for veterinarians (5%; P < 0.001). The most frequently reported injury resulted from exposure to disinfectant and the next was abrasions caused by friction from protective clothing and rubber boots. Among veterinarians, needlestick accidents were also frequently reported. CONCLUSIONS: Applying a less hazardous disinfectant and improving protective equipment to prevent skin abrasion may be effective in reducing the risk of these injuries among staff conducting containment measures. Reducing injuries among staff in such circumstances will assist effective control measures.

Utility of risk-weighted surgical-pathological factors in early-stage cervical cancer.

BACKGROUND: Surgical-pathological risk factors were evaluated by weighting the magnitude of significance of multiple risk factors correlating to survival and treatment response in cervical cancer. METHODS: Multivariate analysis was performed for survival outcomes entering seven pathological factors obtained from 540 radical hysterectomy specimens in stage IA2-IIB cervical cancer cases. Hazard ratio (HR) in each risk factor was determined, and the sum of HR scores for the corresponding risk factors was determined per case. Survival curves and postoperative treatment response (concurrent chemoradiotherapy (CCRT) vs radiotherapy alone) were evaluated based on the extent of HR-weighted scores. RESULTS: Hazard ratios for risk factors relating to disease-free survival (DFS) was: lympho-vascular space invasion 3.95, nodal metastasis 3.88, adenocarcinoma 3.40, large tumour 2.36, positive margin 1.99, deep stromal invasion 1.29, and parametria invasion 1.21. The HR-weighted scoring method showed a high predictive value for recurrence (area-under-curve 0.836, P<0.001). Hazard ratio-weighted scores were negatively correlated to DFS, and the cases with score 12.5 showed 5-year DFS rate of 23.8%. Tumours with larger score offset the benefits of CCRT over radiotherapy alone for postoperative adjuvant treatment (P<0.001). CONCLUSION: Surgical-pathological risk factors provide valuable information for survival and management of early-stage cervical cancer when number and significance of risks are weighted.

Prevalence and growth kinetics of Shiga toxin-producing Escherichia coli (STEC) in bovine offal products in Japan.

Recent epidemiological data suggest a link between the consumption of bovine offal products and Shiga toxin-producing Escherichia coli (STEC) infection in Japan. This study thus examined the prevalence of STEC in various types of these foods. PCR screened 229 bovine offal products for the presence of Shiga toxin (stx) gene. Thirty-eight (16·6%) samples were stx positive, of which eight were positive for rfbE(O157) and three were positive for wzy(O26). Four O157 and one O26 STEC isolates were finally obtained from small-intestine and omasum products. Notably, homogenates of bovine intestinal products significantly reduced the extent of growth of O157 in the enrichment process compared to homogenates of beef carcass. As co-incubation of O157 with background microbiota complex from bovine intestinal products in buffered peptone water, in the absence of meat samples, tended to reduce the extent of growth of O157, we reasoned that certain microbiota present in offal products played a role. In support of this, inoculation of generic E. coli from bovine intestinal products into the homogenates significantly reduced the extent of growth of O157 in the homogenates of bovine intestinal and loin-beef products, and this effect was markedly increased when these homogenates were heat-treated prior to inoculation. Together, this report provides first evidence of the prevalence of STEC in a variety of bovine offal products in Japan. The prevalence data herein may be useful for risk assessment of those products as a potential source of human STEC infection beyond the epidemiological background. The growth characteristic of STEC O157 in offal products also indicates the importance of being aware when to test these food products.

Ovulation compensatory function after unilateral ovariectomy in dogs.

As a step towards elucidation of the timing and mechanism of the determination of the number of ovulated ova in dogs, we excised one ovary 2, 5 and 8 days after the beginning of vulval bleeding and examined whether the lost ovulation function, assessed by estimating the number of ovulated oocytes, would be compensated for by the remaining ovary. The number of ovulated ova was maintained by the remaining ovary in the group that underwent unilateral ovariectomy 2 days after the beginning of vulval bleeding. However, in the groups ovariectomized 5 or 8 days after the beginning of vulval bleeding, no compensation for the number of ova that would have been ovulated from the lost ovary was observed; ova were ovulated only from the follicles 3 mm or greater in diameter observed in the remaining ovary at unilateral ovariectomy. Thus, in dogs, the number of ovulated ova is considered to be determined within 5 days after the beginning of vulval bleeding.

Ovulation day after onset of vulval bleeding in a beagle colony.

This study investigated the duration of the interval between the onset of vulval bleeding at pro-oestrus and ovulation estimated from the plasma progesterone concentration in a large number of beagle bitches. The influence and association of individual variation, ageing and duration of the oestrous cycle were also investigated. The mean time of ovulation after the onset of vulval bleeding was 11.1 ± 0.2 days, but it widely ranged from 3 to 31 days. This timing was not influenced by age or duration of the oestrous cycle, and within-individual variation was small. As there has been no previous report in which the ovulation day was investigated by the age, these data may be very valuable.

Spermatogenic function in cats.

There are limited data on feline sperm production. We exhausted epididymal spermatozoa (i.e. the number of ejaculated spermatozoa <5 × 10(6)) by frequent semen collections using the artificial vagina method in five tomcats and determined the number of spermatozoa stored in the epididymis. We investigated the time (days) required for the number of epididymal spermatozoa to return to the pre-exhaustion level and determined the number of spermatozoa produced per day. After spermatozoa were exhausted by frequent semen collection, 6 or more days were required to return to the pre-exhaustion level. Based on the duration of resting (days) and total number of spermatozoa, the mean number of spermatozoa produced per day was 30 × 10(6).

Simulating the bovine spongiform encephalopathy situation in Japan.

Despite various measures taken by the Japanese government to protect the cattle population from exposure to the bovine spongiform encephalopathy (BSE) agent, the first case of BSE was detected in September 2001. Subsequently, BSE surveillance was enhanced, involving mandatory reporting and investigation of all clinical BSE suspects, and testing of fallen stock and all cattle slaughtered for human consumption. Tests on over nine million cattle led to the detection of 35 additional cases by the end of May 2009. Using the surveillance data and other information as input variables, models were developed to explore the possible source of introduction of BSE into Japan, evaluate the effectiveness of control measures, estimate the prevalence of BSE in different birth cohorts, predict a future BSE epidemic, and simulate the impact of changes in surveillance strategies. Despite difficulties associated with the availability and uncertainty of some of the input variables, these models provided an objective insight into the BSE situation in Japan.

Development of a semi-quantitative evaluation system for surveillance of bovine spongiform encephalopathy, using the analytic hierarchy process.

Evaluating surveillance results is important in estimating the risk of bovine spongiform encephalopathy (BSE) cases in a particular country, or in countries from which products are imported. Although various methods have been proposed for quantitative evaluation of surveillance results, no methods focusing on surveillance from a qualitative perspective have yet been established. The authors have developed an objective method for evaluating the qualitative aspects of BSE surveillance, based on the analytic hierarchy process. Factors related to surveillance credibility were selected through expert meetings and arranged in a hierarchical structure. These evaluation factors were also weighted, so that a points system could be used for evaluation. As a result, 13 evaluation factors comprising three-layer hierarchies were generated. When surveillance in Japan before and after a BSE case was evaluated using this evaluation system, an improvement in the quality of the surveillance was observed after the outbreak. Although this study suggests that the selection of the experts had a significant effect on the outcome, the authors believe that this method will also be applicable for establishing qualitative evaluation systems for other diseases.

Plasma hormone levels and semen quality in male cats during non-breeding and breeding seasons.

Female cats are known to be seasonal breeders and male cats annual breeders. Despite this, there are limited data on the influence of breeding season (BS) on hormone concentration and semen quality in the male cat. This study compared plasma concentrations of LH and testosterone (T), and semen quality during the non-breeding season (NBS) and BS in five male cats subject to natural hours of daylight but a constant environmental temperature. Plasma LH and T concentrations were higher during the BS in 2/35 and 3/5 cats, respectively, although when comparing both hormones combined, values were higher during the BS than the NBS in all cats (p < 0.01). There were no significant differences in the percentage of abnormal sperm between the cats. Overall, semen quality was superior during the BS with larger semen volume in 2/5, sperm motility in 2/5 and sperm viability in 3/5 cats. Although there was a clear seasonal effect on hormone secretion and semen quality, during the NBS all cats were likely to have been fertile.

Effect of catalase and superoxide dismutase on motility, viability and acrosomal integrity of frozen-thawed cat spermatozoa.

The aim of this study was to evaluate the effect of antioxidant catalase (CAT) and superoxide dismutase (SOD) in semen extender on motility, viability and acrosomal integrity of frozen-thawed cat spermatozoa. Semen was collected by using an artificial vagina from five domestic cats (two ejaculates/cat). Spermatozoa were diluted in egg yolk Ttris-fructose citrate solution (EYT-FC) without glycerol and cooled at 4 degrees C for 1 h, then diluted further with EYT-FC with glycerol (7% final concentration) and 400 IU/ml of CAT (treatment 1) or SOD (treatment 2) or without antioxidants (control). Before freezing using a styrofoam box, diluted spermatozoa filled in 0.25-ml straws were equilibrated for 1 h at 4 degrees C. After thawing, spermatozoa were assessed for motility, viability and acrosomal integrity. Cryopreservation significantly impaired sperm motility, viability and acrosomal integrity (p < 0.05). However, motility, viability and acrosomal integrity of frozen-thawed cat spermatozoa in the EYT-FC with CAT, SOD and without the antioxidants were not significantly different. The average percentages of spermatozoa motility after thawing compared between control, treatment 1 and treatment 2 group were 43.5 +/- 3.2, 42 +/- 4.1 and 38 +/- 4.5; for viability: 44.8 +/- 3.5, 50.6 +/- 5.7 and 47.1 +/- 4.1 and for acrosomal integrity: 45 +/- 3.5, 44.9 +/- 3.4 and 44.4 +/- 3.3, respectively. In conclusion, adding CAT and SOD to EYT-FC did not improve motility, viability and acrosomal integrity in cryopreserved cat spermatozoa.

Plasma LH, ovulation and conception rates in cats mated once or three times on different days of oestrus.

Although cats are induced ovulators, the relationship between the day of breeding, the number of matings and the likelihood of ovulation and conception have not been extensively investigated. In this experiment, cats were mated either once or three times on day 1 or day 5 of oestrus to study the incidence of the LH surge, ovulation and conception rates. The percentage ovulating and the conception rates after a single mating on day 1 of oestrus were 60% (6/10) and 33.3% (2/6), respectively, and for cats mated once on day 5 of oestrus were 83.3% (10/12) and 40% (4/10), respectively. When cats were mated three times on day 1 of oestrus, the ovulation rates and conception rates were 70% (7/10) and 85.7% (6/7), respectively, and for those mated three times on day 5 of oestrus were 100% (10/10) and 100% (10/10), respectively. The concentration of LH did not increase in non-ovulating cats, and cats that were mated three times had LH concentrations that were numerically higher than those that were mated once. Litter size was neither related to the day of mating nor to the number of matings. Although an increase in the number of matings on day 1 of oestrus produced a numerically larger LH surge, it did not increase the ovulation rate, suggesting that plasma oestradiol concentrations were not sufficiently elevated to induce a high pituitary response to mating stimulation. The conception rate after a single mating was low, suggesting that the number of sperm per mating was not sufficient. These results suggest that mating more than once in the middle of oestrus is required to improve ovulation rates and conception rates in cats.

The role of the ovary for the maintenance of pregnancy in cats.

Plasma progesterone (P(4)) concentrations are maintained in pregnant cats until parturition, but become low in pseudopregnant cats 40-45 days after infertile mating. This difference in P(4) concentrations is considered to be due to P(4) secretion by the placenta of pregnant cats. Therefore, to clarify these points, we performed ovariectomy (OVX) at various stages of pregnancy, examined the pregnancy status and measured LH and P(4) concentrations in peripheral, ovarian and uterine venous blood. After OVX, abortion occurred in 100% (5/5), 80% (4/5), 40% (2/5) and 60% (3/5) of Groups I (Day 35), II (Day 40), III (Day 45) and IV (Day 50) cats, respectively. In the remaining cats, normal delivery took place on days 63-69 [mean, 66.1 +/- 1.1 (SE)] of pregnancy. The time to abortion after OVX was 4-8 (mean, 5.6 +/- 0.8), 3-17 (mean, 8.0 +/- 3.6), 10 and 11, and 2-4 (mean, 3.0 +/- 0.7) days in Groups I, II, III and IV, respectively. The plasma P(4) concentrations were 1-2 ng/ml in all groups on the day after OVX, decreasing to less than 1 ng/ml from the 2nd day onwards. The concentrations of P(4) in ovarian venous blood at the time of OVX decreased with the stage of pregnancy, but were clearly higher than those in peripheral blood. The plasma P(4) concentrations in uterine venous blood were similar to those in peripheral blood. These results suggest that peripheral P(4) in pregnant cats is the result of P(4) secretion secreted only by the ovarian corpus luteum, not by the placenta, but indicate that either P(4) is not essential for the maintenance of pregnancy in cats from day 40-45 of pregnancy onwards, or that the placenta provides a local source of P(4) that does not appear in measurable amounts in the peripheral circulation.

Prolonged duration of fertility of dog ova.

The fertile period for natural mating in dogs extends from before ovulation until day 5 post ovulation (PO) and involves a delay in oocyte maturation until 2-3 days PO and viability of secondary oocytes for 48-60 h or more. Spermatozoa do not enter the uterus after vaginal insemination in late oestrus. Cervical closure appears to occur on average 5 days PO, but conception may occur following intrauterine artificial insemination (IUAI) up to 8 days PO. Therefore, the present study was conducted to clarify the duration of fertility of canine ova. Using IUAI at 6, 7, 8 and 9 days PO (n = 5 bitches each) conception rates were 100%, 71.4%, 37.5% and 0%, respectively, with an average litter resorption rate of 30.8%, and with mean litter sizes and times to delivery PO being 4.3 +/- 1.6 and 64.3 +/- 0.3 days, 4.0 +/- 1.4 and 66.3 +/- 0.4 days, and 2.5 and 68 days for IUAI at 6, 7 and 8 days, respectively. The high pregnancy rates with IUAI at 6 and 7 days PO confirm that many canine oocytes are fertile at 4-5 days after maturation. The high rate of resorption was presumably because of aging of ova or asynchrony between embryonic development and the intrauterine environment.

Development of craniofacial structures in transgenic mice with constitutively active PTH/PTHrP receptor.

Parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP) regulate calcium homeostasis, and PTHrP further regulates growth and development. A transgenic mouse carrying the constitutively active PTH/PTHrP receptor (HKrk-H223R) under the control of the mouse bone and odontoblast-specific alpha1(I) collagen promoter (Col1-caPPR) has been developed to demonstrate the complex actions of this mutant receptor in hard tissue formation. We have further characterized Col1-caPPR mice abnormalities in the craniofacial region as a function of development. Col1-caPPR mice exhibited a delay in embryonic bone formation, followed by expansion of a number of craniofacial bones including the maxilla and mandible, delay in tooth eruption and teratosis, and a disrupted temporomandibular joint (TMJ). These findings suggest that the Col1-caPPR mouse is a useful model for characterization of the downstream effects of the constitutively active receptor during development and growth, and as a model for development of treatments of human diseases with similar characteristics.

Transition of Akabane virus genogroups and its association with changes in the nature of disease in Japan.

Akabane virus (AKAV) is teratogenic to the foetus of domestic ruminants and causes a significant reproduction loss in cattle in Japan. In several past epizootics in cattle, AKAV was also associated with post-natal encephalomyelitis, mainly in calves and young stock. Previously analysed AKAV isolates in East Asia form two major clusters, genogroups I and II, with isolates involved in encephalomyelitis belonging mainly to the former. Between 2007 and 2013, AKAV epizootics were regularly observed in Japan during the summer/autumn season, and abnormal deliveries and post-natal encephalomyelitis caused by the virus in cattle were reported. During this period, 30 AKAV isolates were obtained from diseased and sentinel cattle, a piglet and Culicoides biting midges throughout Japan and were subjected to genetic comparison and phylogenetic analysis with previous isolates. In 2007, 2011 and 2013, AKAV belonging to genogroup I was identified in the central nervous systems of calves showing neurological disorders. Notably, a total of 165 cases of bovine encephalomyelitis were reported in 2011 and the isolated viruses from affected animals shared high genetic identities with a South Korean isolate that was associated with a large outbreak in 2010, suggesting some epidemiological linkage between these epizootics. Epizootics of genogroup II were observed in 2008 and 2010, but bovine post-natal encephalomyelitis cases rarely occurred. Our findings suggest that the frequent incursion of genogroup I isolates has increased the frequency of post-natal encephalomyelitis cases in Japan in recent years. Infection by genogroup I virus was also identified in piglets with neurological disorders or congenital malformations in 2011 and 2013. The aetiological role of AKAV in pigs should be elucidated in the future.

Targeted disruption of CDK4 delays cell cycle entry with enhanced p27(Kip1) activity.

The mechanism by which cyclin-dependent kinase 4 (CDK4) regulates cell cycle progression is not entirely clear. Cyclin D/CDK4 appears to initiate phosphorylation of retinoblastoma protein (Rb) leading to inactivation of the S-phase-inhibitory action of Rb. However, cyclin D/CDK4 has been postulated to act in a noncatalytic manner to regulate the cyclin E/CDK2-inhibitory activity of p27(Kip1) by sequestration. In this study we investigated the roles of CDK4 in cell cycle regulation by targeted disruption of the mouse CDK4 gene. CDK4(-/-) mice survived embryogenesis and showed growth retardation and reproductive dysfunction associated with hypoplastic seminiferous tubules in the testis and perturbed corpus luteum formation in the ovary. These phenotypes appear to be opposite to those of p27-deficient mice such as gigantism and gonadal hyperplasia. A majority of CDK4(-/-) mice developed diabetes mellitus by 6 weeks, associated with degeneration of pancreatic islets. Fibroblasts from CDK4(-/-) mouse embryos proliferated similarly to wild-type embryonic fibroblasts under conditions that promote continuous growth. However, quiescent CDK4(-/-) fibroblasts exhibited a substantial ( approximately 6-h) delay in S-phase entry after serum stimulation. This cell cycle perturbation by CDK4 disruption was associated with increased binding of p27 to cyclin E/CDK2 and diminished activation of CDK2 accompanied by impaired Rb phosphorylation. Importantly, fibroblasts from CDK4(-/-) p27(-/-) embryos displayed partially restored kinetics of the G(0)-S transition, indicating the significance of the sequestration of p27 by CDK4. These results suggest that at least part of CDK4's participation in the rate-limiting mechanism for the G(0)-S transition consists of controlling p27 activity.

The cell cycle-regulatory CDC25A phosphatase inhibits apoptosis signal-regulating kinase 1.

CDC25A phosphatase promotes cell cycle progression by activating G(1) cyclin-dependent kinases and has been postulated to be an oncogene because of its ability to cooperate with RAS to transform rodent fibroblasts. In this study, we have identified apoptosis signal-regulating kinase 1 (ASK1) as a CDC25A-interacting protein by yeast two-hybrid screening. ASK1 activates the p38 mitogen-activated protein kinase (MAPK) and c-Jun NH(2)-terminal protein kinase-stress-activated protein kinase (JNK/SAPK) pathways upon various cellular stresses. Coimmunoprecipitation studies demonstrated that CDC25A physically associates with ASK1 in mammalian cells, and immunocytochemistry with confocal laser-scanning microscopy showed that these two proteins colocalize in the cytoplasm. The carboxyl terminus of CDC25A binds to a domain of ASK1 adjacent to its kinase domain and inhibits the kinase activity of ASK1, independent of and without effect on the phosphatase activity of CDC25A. This inhibitory action of CDC25A on ASK1 activity involves diminished homo-oligomerization of ASK1. Increased cellular expression of wild-type or phosphatase-inactive CDC25A from inducible transgenes suppresses oxidant-dependent activation of ASK1, p38, and JNK1 and reduces specific sensitivity to cell death triggered by oxidative stress, but not other apoptotic stimuli. Thus, increased expression of CDC25A, frequently observed in human cancers, could contribute to reduced cellular responsiveness to oxidative stress under mitogenic or oncogenic conditions, while it promotes cell cycle progression. These observations propose a mechanism of oncogenic transformation by the dual function of CDC25A on cell cycle progression and stress responses.