Foramen magnum stenosis and midface hypoplasia in C-type natriuretic peptide-deficient rats and restoration by the administration of human C-type natriuretic peptide with 53 amino acids.

C-type natriuretic peptide (CNP)-knockout (KO) rats exhibit impaired skeletal growth, with long bones shorter than those in wild-type (WT) rats. This study compared craniofacial morphology in the CNP-KO rat with that in the Spontaneous Dwarf Rat (SDR), a growth hormone (GH)-deficient model. The effects of subcutaneous administration of human CNP with 53 amino acids (CNP-53) from 5 weeks of age for 4 weeks on craniofacial morphology in CNP-KO rats were also investigated. Skulls of CNP-KO rats at 9 weeks of age were longitudinally shorter and the foramen magnum was smaller than WT rats. There were no differences in foramen magnum stenosis and midface hypoplasia between CNP-KO rats at 9 and 33 weeks of age. These morphological features were the same as those observed in CNP-KO mice and activated fibroblast growth factor receptor 3 achondroplasia-phenotype mice. In contrast, SDR did not exhibit foramen magnum stenosis and midface hypoplasia, despite shorter stature than in control rats. After administration of exogenous CNP-53, the longitudinal skull length and foramen magnum size in CNP-KO rats were significantly greater, and full or partial rescue was confirmed. The synchondrosis at the cranial base in CNP-KO rats is closed at 9 weeks, but not at 4 weeks of age. In contrast, synchondrosis closure in CNP-KO rats treated with CNP-53 was incomplete at 9 weeks of age. Administration of exogenous CNP-53 accelerated craniofacial skeletogenesis, leading to improvement in craniofacial morphology. As these findings in CNP-KO rats are similar to those in patients with achondroplasia, treatment with CNP-53 or a CNP analog may be able to restore craniofacial morphology and foramen magnum size as well as short stature.

Incidence of venous thromboembolism and bleeding events in patients with lower extremity orthopedic surgery: a retrospective analysis of a Japanese healthcare database.

BACKGROUND: Orthopedic surgeries of lower extremities such as total knee arthroplasty (TKA), total hip arthroplasty (THA), and hip fracture surgery (HFS) are widely considered to carry a high risk of developing deep venous thrombosis (DVT) and pulmonary thromboembolism (PTE). Growing attention to epidemiological studies using a healthcare database led us to quantify the risks using a Japanese database to reveal recent medical care for such events. METHODS: The study comprised 36,947 patients who had undergone orthopedic surgeries of the lower extremities and whose medical information from April 2008 to September 2013 was available. The source population of the database was derived from 100 acute-care hospitals with Diagnosis Procedure Combination. The events were defined by diagnosis, medication, imaging, and laboratory tests. RESULTS: A breakdown of patients who underwent orthopedic surgeries by type of surgery showed 13.6% for TKA, 10.4% for THA, 56.8% for HFS, 1.5% for rupture of Achilles tendon, and 18.0% for simple fracture of lower extremities. The incidence for DVT, PTE, and bleeding events by type of surgery was 1.3, 0.2, and 1.0% for TKA; 0.9, 0.2, and 1.1% for THA; and 0.4, 0.1, and 1.8% for HFS, respectively. The population for risk factor analysis consisted of patients with similar background factors who underwent TKA, THA, or HFS. The statistically significant risk factors for PTE analyzed by the backward elimination procedure in a multivariate model were female sex, history of venous thromboembolism (VTE), thrombophilia, and varicose veins of lower extremity. CONCLUSIONS: The incidence of DVT, PTE, and bleeding events and the risk factors for DVT and PTE in patients by type of orthopedic surgeries of lower extremities found in our study are considered to be rational as they reflect evidence from real-world cases. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry UMIN000012667.

A comprehensive dose evaluation project concerning animals affected by the Fukushima Daiichi Nuclear Power Plant accident: its set-up and progress.

It is not an exaggeration to say that, without nuclear accidents or the analysis of radiation therapy, there is no way in which we are able to quantify radiation effects on humans. Therefore, the livestock abandoned in the ex-evacuation zone and euthanized due to the Fukushima Daiichi Nuclear Power Plant (FNPP) accident are extremely valuable for analyzing the environmental pollution, its biodistribution, the metabolism of radionuclides, dose evaluation and the influence of internal exposure. We, therefore, sought to establish an archive system and to open it to researchers for increasing our understanding of radiation biology and improving protection against radiation. The sample bank of animals affected by the FNPP accident consists of frozen tissue samples, formalin-fixed paraffin-embedded specimens, dose of radionuclides deposited, etc., with individual sampling data.

A randomized, open-label trial of edoxaban in Japanese patients with severe renal impairment undergoing lower-limb orthopedic surgery.

BACKGROUND: Edoxaban is an oral, direct, factor Xa inhibitor approved in Japan for thromboembolic prophylaxis after lower-limb orthopedic surgery (LLOS), but contraindicated in patients with severe renal impairment (SRI; creatinine clearance [CLCR] ≥15 to <30 mL/min). METHODS: This open-label study compared the safety of edoxaban 15 mg once daily in Japanese patients with SRI to that of edoxaban 30 mg in patients with mild renal impairment (MiRI; CLCR ≥50 to ≤80 mL/min; N = 30) undergoing LLOS. Patients with CLCR ≥20 to <30 mL/min were randomized to receive edoxaban 15 mg (N = 22) or subcutaneous fondaparinux 1.5 mg once daily (N = 21). All patients with CLCR ≥15 to <20 mL/min received edoxaban 15 mg (N = 7). Treatment was administered for 11 to 14 days. RESULTS: Major or clinically relevant non-major bleeding occurred in 6.7%, 3.4%, and 5.0% of patients in the MiRI edoxaban 30-mg, SRI edoxaban 15-mg, and SRI fondaparinux groups, respectively; there were no major bleeding events. No thromboembolic events occurred. At all time points assessed, edoxaban plasma concentrations and changes in coagulation biomarkers were similar between the SRI and MiRI groups. CONCLUSIONS: These results suggest edoxaban 15 mg once daily is well tolerated in Japanese patients with SRI undergoing LLOS. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT01857583.

Bone morphogenetic protein 4 (BMP-4) and BMP-7 induce vascular endothelial growth factor expression in bovine granulosa cells.

Bone morphogenetic protein-4 (BMP-4) and BMP-7, theca cell-derived growth factors, directly affect the granulosa cell function. The aim of this study was to examine the involvement of BMP-4 or BMP-7 in vascular endothelial growth factor (VEGF) expression in bovine granulosa cells. Granulosa cells were collected from small follicles (4-6 mm) and seeded at a density of 2-5 × 10(5) cells per well in Dulbecco's modified Eagle's medium (DMEM)/F12 medium with BMP-4 or BMP-7. The expression of VEGF messenger RNA and protein was the maximum when 1.0 ng/mL of BMP-4 was added to the culture medium. On the other hand, 10 ng/mL of BMP-7 significantly increased the expression of the VEGF gene and protein. In addition, BMP-4 stimulated the expression of Smad1 and Smad5 genes in granulosa cells, whereas BMP-7 stimulated the expression of Smad5 gene. These results suggested that BMP-4 and BMP-7 may be associated with VEGF expression via several specific Smads in bovine granulosa cells: BMP-4 via Smad1/Smad5 and BMP-7 via Smad5. In conclusion, theca cell-derived BMP-4 and BMP-7 might contribute to follicular vasculature and development by inducing VEGF expression in granulosa cells.

Cryopreservation of canine embryos.

The assisted reproductive techniques (ARTs) such as in vitro fertilization, embryo transfer, and cryopreservation of gametes have contributed considerably to the development of biomedical sciences in addition to improving infertility treatments in humans as well as the breeding of domestic animals. However, ARTs used in canine species have strictly limited utility when compared with other mammalian species, including humans. Although successful somatic cell cloning has been reported, artificial insemination by frozen semen to date is only available for the improved breeding and reproduction for companion and working dogs as well as guide dogs for the blind. We describe here the successful cryopreservation of embryos and subsequent embryo transfer in dogs. Canine embryos were collected from excised reproductive organs after artificial insemination and subsequently cryopreserved by a vitrification method. When the 4-cell to morula stage of cryopreserved embryos were nonsurgically transferred into the uteri of nine recipient bitches using a cystoscope, five recipients became pregnant and four of them delivered a total of seven pups. The cryopreservation of embryos in canine species will facilitate the transportation and storage of genetic materials and will aid in the elimination of vertically transmitted diseases in dogs. In addition, this technique will contribute to the improved breeding of companion and working dogs such as guide dogs, drug-detecting dogs, and quarantine dogs.

Effect of graft site and gonadotrophin treatment on follicular development of canine ovarian grafts transplanted to NOD-SCID mice.

PURPOSE: To investigate the effect of graft site and gonadotrophins administration on the number and survival rate of follicles of canine ovarian grafts transplanted to NOD-severe combined immune deficiency (SCID) mice. METHODS: Fresh ovarian cortex slices obtained from immature bitches were grafted subcutaneously (SC), under kidney capsule (KC) or into ovarian bursa (OB) in NOD-SCID mice. Two months after surgery, the mice allocated into non-treated and treated gonadotrophins groups that injected with porcine follicle stimulating hormone during 7 days and human chorionic gonadotrophin 48 h later. Ovarian grafts were collected after 10 h of last injection and processed for histology. RESULTS: The number of transitional and preantral follicles under KC and into OB was significantly higher in gonadotrophins-treated mice than those who received saline. Furthermore, the survival rates of primary, transitional and preantral follicles under KC and into OB grafts were significantly higher than those placed SC in the treated gonadotrophins group, and in the non-treated gonadotrophins group; the proportion of primary and preantral follicle survival was significantly higher under KC and into OB than SC grafts. CONCLUSIONS: In canine ovarian xenografting, administration of gonadotrophin could be effective for improvement of survival of transplanted ovary. Furthermore, the grafting into OB appeared to be better than grafting under KC, which in turn is better than SC.

Prediction of postoperative ambulatory status 1 year after hip fracture surgery.

UNLABELLED: Hirose J, Ide J, Yakushiji T, Abe Y, Nishida K, Maeda S, Anraku Y, Usuku K, Mizuta H. Prediction of postoperative ambulatory status 1 year after hip fracture surgery. OBJECTIVES: To assess the validity of Estimation of Physiologic Ability and Surgical Stress (E-PASS) for predicting the postoperative risk and ambulatory status long-term follow-up after hip fracture surgery and to establish an algorithm for predicting their ambulatory status. DESIGN: Cohort study. SETTING: Twelve hospitals belonging to the regional network for hip fracture in Japan. PARTICIPANTS: The study population was composed of 421 patients; 268 underwent surgery between April 2004 and March 2006 (group A), and 153 were treated surgically between April 2006 and March 2007 (group B). All were operated at 3 surgical hospitals and, subsequently, transferred to 9 rehabilitation centers. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: We evaluated various factors, including their E-PASS scores to determine whether there was a correlation with the patients' mortality rate and their ability to walk at discharge and 1 year after surgery (group A). Using multiple regression analysis, we then developed algorithms to predict the ability of elderly patients to walk after hip fracture surgery. We applied the algorithms to group B patients and compared their actual and predicted ambulatory status. RESULTS: In group A patients, the postoperative walking ability and mortality rate were highly correlated with their E-PASS scores and dementia status. In group B, our algorithms exhibited good correlations between the predicted and actual walking ability at both time points (rho=0.6, P<.001). CONCLUSIONS: In candidates for hip fracture surgery, the E-PASS scores exhibited a good correlation with the patients' functional and survival prognoses, and the algorithm including E-PASS scores and dementia status can accurately estimate the ambulatory status at discharge and 1 year after surgery.

Individual fertility differences in the frozen-thawed spermatozoa among semen donors in the Labrador Retriever.

Purpose: We aimed to validate and determine the possible application of transcervical insemination of frozen semen for improved breeding in guide dogs for the blind in Japan. Methods: From February 2004 to March 2007, a total of 53 Labrador Retriever bitches, used for the breeding of guide dogs for the blind, were transcervically inseminated with frozen-thawed semen from 13 males by means of a cystoscope. Results: The overall whelping and pregnancy rate with the frozen semen was 42%. Pregnancy rates ranged widely from 0 to 100% depending on the semen donor male. Of 13 males, 6 males exhibited severely poor fertility (less than 20% pregnancy rate) and 3 males exhibited high fertility (over 70% pregnancy rate) on artificial insemination. However, the spermatozoa motility after thawing was not significantly different among these dogs. In addition, heterospermic insemination revealed the optimal timing for transcervical insemination with frozen-thawed semen to be by day 6 after the LH surge. Conclusions: Although transcervical insemination of frozen-thawed semen is effective for breeding of guide dogs for the blind, some modification of freeze-thawing procedures might be required to overcome individual fertility differences in the frozen-thawed spermatozoa among semen donor dogs,. In addition, the motility of spermatozoa after thawing might not be an appropriate indicator of the relative fertility of frozen-thawed spermatozoa in dogs.

Recent patents of TGF-beta family and VEGF associated with ovarian follicular development in mammals.

In mammalian ovary, follicular development in mammals is regulated by the complex process including endocrine, paracrine and autocrine. During the last decade, the role of growth factors in ovarian folliculogenesis has been extensively studied in mammals. In particular, a growing body of evidence indicates that the vascular endothelial growth factor (VEGF) system plays a key role in follicular development and atresia in the woman, rodents and domestic animal species. More recently, the bone morphogenetic protein (BMP) and growth differentiation factor (GDF) that belong to TGF-beta family have been shown to be involved in the regulation of follicle growth. In this paper, we will essentially consider the role of these growth factor systems in mammalian ovary. Moreover, we will review recent patents associated with ovarian follicular development in mammals.

Expression of Pin1, a peptidyl-prolyl isomerase, in the ovaries of eCG/hCG-treated immature female mice.

Protein phosphorylation on certain serine or threonine residues preceding proline (Ser/Thr-Pro) is a pivotal signaling mechanism in diverse cellular processes. Pin1 is a highly conserved enzyme that isomerizes only the phosphorylated Ser/Thr-Pro bonds in certain proteins, thereby inducing conformational changes. Although much protein is phosphorylated in the ovary, the role of Pin1 in the ovary is still unknown. The purpose of this study is to investigate the effects of gonadotropins on protein and mRNA expression of Pin1 in mice ovaries. Quantitative PCR analysis showed that the expression of Pin1 mRNA significantly increased in the ovaries of equine chorionic gonadotropin (eCG)-treated mice compared with those of untreated mice (P<0.05). However, human chorionic gonadotropin (hCG) attenuated the expression of Pin1 mRNA increased by eCG. The protein level of Pin1 showed the same tendency as the expression of mRNA. The mRNA expression of E2F transcription factor, which controlled the expression of Pin1, was significantly decreased in the eCG-treated ovaries compared with the controls (P<0.05). These observations suggest that gonadotropins may regulate the expression of Pin1 without E2F transcription factor, indicating that Pin1 might be an important factor for protein signal transduction during follicular development.

Bax-inhibiting peptide derived from mouse and rat Ku70.

Bax is a proapoptotic protein that plays a key role in the induction of apoptosis. Ku70 has activities to repair DNA damage in the nucleus and to suppress apoptosis by inhibiting Bax in the cytosol. We previously designed peptides based on the amino acid sequence of Bax-binding domain of human Ku70, and showed that these peptides bind Bax and inhibit cell death in human cell lines. In the present report, we examined the biological activities of other pentapeptides, VPTLK and VPALR, derived from mouse and rat Ku70. Cells in culture accumulated FITC-labeled VPTLK and VPALR, indicating that these peptides are cell permeable (human, mouse, rat, and porcine cells were examined). These peptides bound to Bax and suppressed cell death in various cell types including primary cultured cells. These data suggest that such Bax inhibiting peptides from three mammalian species may be used to protect healthy cells from apoptotic injury under pathological conditions.

Primary clear cell sarcoma (malignant melanoma) in the right radius.

Clear cell sarcoma (malignant melanoma) originating in bone is an extremely rare occurrence, which has been reported twice previously. It is challenging to differentiate this neoplasm from skeletal metastasis of malignant melanoma because it shows no specific imaging, pathological or immunohistochemical features. However, this differentiation is clinically important due to significant differences in patient management. In this article, we present the case of a 55-year-old man with primary clear cell sarcoma arising in the right radius.

Follicular microvasculature and angiogenic factors in the ovaries of domestic animals.

The genetic and molecular mechanisms that control the development of capillary blood vessels during follicular development are beginning to be elucidated. Ovarian follicles contain and produce angiogenic factors that may act alone or in concert to regulate the process of thecal angiogenesis. These factors are ultimately controlled by endocrine, paracrine and autocrine regulation. A recent study indicated that vascular endothelial growth factor (VEGF) plays an important role in the process of thecal angiogenesis during follicular development. We are developing a novel technology for the induction of follicular development using the technique of in vivo gene administration. Here, we summarize the recent progress of our research.