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Anaerobic threshold (AT) from cardiopulmonary exercise tests (CPX) is the standard for measuring exercise intensity among patients with cardiovascular disease in Japan. However, it remains controversial whether AT represents the safety limit for exercise intensity in patients with cardiovascular disease. The purpose of this study was to investigate cardiac rehabilitation (CR) efficacy and safety with exercise intensities above the AT and at a traditional AT in a randomized trial. The participants included 57 patients who were admitted to the outpatient CR unit with a diagnosis of acute myocardial infarction. The participants were randomly divided as follows: 25 patients in the AT group, who performed aerobic exercises with an intensity at the AT; and 32 patients in the "Over AT" group, who performed exercises at an intensity higher than the AT. The following components were measured: maximum oxygen uptake (peak VO 2 ), oxygen uptake at the AT (AT VO 2 ), increase in oxygen uptake during exercise (ΔVO 2 /ΔWR) during the CPX, vascular endothelial function test (%FMD: the percentage of flow-mediated dilation), and isometric knee extension strength. The measurements were obtained at the start of the exercise therapy and after 2, 3, and 4 months. They were compared within and between groups, and the correlation between the rates of improvement was investigated. Peak VO 2 , AT VO 2 , ΔVO 2 /ΔWR, and %FMD had significantly improved after 3 months in both groups. The isometric knee extension strength had improved in the "Over AT" group after 2 months. Interactions were observed with peak VO 2 , ΔVO 2 /ΔWR, and isometric knee extension strength. However, %FMD was not significantly different between the groups. In the "Over AT" group, the rate of improvement in peak VO 2 was positively correlated with the improvement in the isometric knee extension strength (r = 0.61, p < 0.001), but not with %FMD. These data suggest that exercise at an intensity above the AT improved exercise tolerance faster than that at the AT, and this improvement rate was associated with changes in isometric knee extension strength.

The deployment of a large-scale indoor sensor network faces a sensor localization problem because we need to manually locate significantly large numbers of sensors when Global Positioning System (GPS) is unavailable in an indoor environment. Fingerprinting localization is a popular indoor localization method relying on the received signal strength (RSS) of radio signals, which helps to solve the sensor localization problem. However, fingerprinting suffers from low accuracy because of an RSS instability, particularly in sensor localization, owing to low-power ZigBee modules used on sensor nodes. In this paper, we present MuCHLoc, a fingerprinting sensor localization system that improves the localization accuracy by utilizing channel diversity. The key idea of MuCHLoc is the extraction of channel diversity from the RSS of Wi-Fi access points (APs) measured on multiple ZigBee channels through fingerprinting localization. MuCHLoc overcomes the RSS instability by increasing the dimensions of the fingerprints using channel diversity. We conducted experiments collecting the RSS of Wi-Fi APs in a practical environment while switching the ZigBee channels, and evaluated the localization accuracy. The evaluations revealed that MuCHLoc improves the localization accuracy by approximately 15% compared to localization using a single channel. We also showed that MuCHLoc is effective in a dynamic radio environment where the radio propagation channel is unstable from the movement of objects including humans.

A bathroom has higher probability of accidents than other rooms due to a slippery floor and temperature change. Because of high privacy and humidity, we face difficulties in monitoring inside a bathroom using traditional healthcare methods based on cameras and wearable sensors. In this paper, we present a danger-pose detection system using commodity Wi-Fi devices, which can be applied to bathroom monitoring, preserving privacy. A machine learning-based detection method usually requires data collected in target situations, which is difficult in detection-of-danger situations. We therefore employ a machine learning-based anomaly-detection method that requires a small amount of data in anomaly conditions, minimizing the required training data collected in dangerous conditions. We first derive the amplitude and phase shift from Wi-Fi channel state information (CSI) to extract low-frequency components that are related to human activities. We then separately extract static and dynamic features from the CSI changes in time. Finally, the static and dynamic features are fed into a one-class support vector machine (SVM), which is used as an anomaly-detection method, to classify whether a user is not in bathtub, bathing safely, or in dangerous conditions. We conducted experimental evaluations and demonstrated that our danger-pose detection system achieved a high detection performance in a non-line-of-sight (NLOS) scenario.

Background: Toe grip strength is the force of a toe on a surface. The objective of this study was to investigate the relationship between falls in middle-aged individuals and physical strength factors such as toe grip strength and knee extension strength., Methods: The subjects were 194 middle-aged individuals (388 feet) who were independent in daily life, received no nursing care, and participated in a health sports event organized by a sports club. We evaluated the body composition, blood pressure, vascular age, systemic response, bone density, knee extension strength, and toe grip strength, and examined their relationship using a self-administered questionnaire survey., Results: The fall, near-fall, and no fall groups included 7, 36, and 151 subjects, respectively; the high and low risk groups included 43 and 151 subjects, respectively. Logistic regression analysis was performed with risk of falls as the dependent variable, and factors that showed a significant difference in the comparison of the high and low risk groups as independent variables. In this analysis, toe grip strength and diastolic blood pressure were identified as independent risk factors for a fall., Conclusions: Toe grip strength is an independent risk factor for falls, and improvement of toe grip strength might prevent falls.

WT4869 is a synthetic peptide vaccine derived from the Wilms' tumor gene 1 (WT1) protein. This phase 1/2 open-label study evaluated the safety and efficacy of WT4869, and biomarkers for response, in patients with myelodysplastic syndrome. WT4869 (5-1200 μg/dose) was administered intradermally every 2 weeks, according to a 3 + 3 dose-escalation method in higher-risk (International Prognostic Scoring System score ≥1.5) or lower-risk (score <1.5) red blood cell transfusion-dependent patients with myelodysplastic syndrome. Twenty-six patients were enrolled and treated (median age, 75 years; range, 32 to 89). The most common adverse event was injection site reaction (61.5%). Main grade 3 or 4 adverse events were neutropenia (30.8%), febrile neutropenia, pneumonia, elevated blood creatine phosphokinase levels and hypoalbuminemia (all 7.7%). Dose-limiting toxicities occurred in 1 patient in the 50 μg/dose cohort (pyrexia, muscle hemorrhage and hypoalbuminemia) and 1 patient in the 400 μg/dose cohort (pneumonitis); however, the maximum tolerated dose could not be determined from this trial. The overall response rate was 18.2%, the disease control rate was 59.1% and median overall survival was 64.71 weeks (95% confidence interval: 50.29, 142.86) as assessed by the Kaplan-Meier method. Subgroup analysis of azacitidine-refractory patients with higher-risk myelodysplastic syndrome (11 patients) showed median overall survival of 55.71 weeks (approximately 13 months). WT1-specific cytotoxic T lymphocyte induction was observed in 11 of 25 evaluable patients. WT4869 was well tolerated in patients with myelodysplastic syndrome and preliminary data suggest that WT4869 is efficacious. This trial was registered at www.clinicaltrials.jp as JapicCTI-101374., (© 2017 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

We aimed to clarify the longitudinal course of mild parkinsonian signs (MPS) and their association with dementia and functional disability by conducting a comprehensive epidemiological study, including brain MRI, and assessments of cognition, depression, and sleep, in people aged ≥65years living in Ama-cho. We diagnosed MPS and parkinsonism (PS) using a modified Unified Parkinson's Disease Rating Scale. The phase I study was conducted between 2008 and 2010 (n=729) and the phase II between 2011 and 2013 (n=436). By phase II, 8.5% of the phase I participants without PS had developed PS. In addition to older age, a lower Mini-Mental State Examination (MMSE) score, and lower body mass index, the MPS rigidity subtype was a significant independent predictor of PS onset. By phase II, 10.1% of the participants without dementia or PS at phase I had developed dementia. Older age, lower MMSE score, and the axial dysfunction and tremor MPS subtypes were significant independent predictors of dementia development. By phase II, 38.8% of participants with MPS at phase I showed no motor symptoms. Younger age and adequate sleep were significant predictors for this reversion. Periventricular and deep white matter hyperintensity Fazekas scores increased with the evolution of parkinsonian signs. MPS is therefore critically, although sometimes reversibly, associated with PS and dementia development in elderly people., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Mps1, also known as TTK, is a dual-specificity kinase that regulates the spindle assembly check point. Increased expression levels of Mps1 are observed in cancer cells, and the expression levels correlate well with tumor grade. Such evidence points to selective inhibition of Mps1 as an attractive strategy for cancer therapeutics. Starting from an aminopyridine-based lead 3a that binds to a flipped-peptide conformation at the hinge region in Mps1, elaboration of the aminopyridine scaffold at the 2- and 6-positions led to the discovery of 19c that exhibited no significant inhibition for 287 kinases as well as improved cellular Mps1 and antiproliferative activities in A549 lung carcinoma cells (cellular Mps1 IC₅₀=5.3 nM, A549 IC₅₀=26 nM). A clear correlation between cellular Mps1 and antiproliferative IC₅₀ values indicated that the antiproliferative activity observed in A549 cells would be responsible for the cellular inhibition of Mps1. The X-ray structure of 19c in complex with Mps1 revealed that this compound retains the ability to bind to the peptide flip conformation. Finally, comparative analysis of the X-ray structures of 19c, a deamino analogue 33, and a known Mps1 inhibitor bound to Mps1 provided insights into the unique binding mode at the hinge region., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Monopolar spindle 1 (Mps1) is an attractive oncology target due to its high expression level in cancer cells as well as the correlation of its expression levels with histological grades of cancers. An imidazo[1,2-a]pyrazine 10a was identified during an HTS campaign. Although 10a exhibited good biochemical activity, its moderate cellular as well as antiproliferative activities needed to be improved. The cocrystal structure of an analogue of 10a guided our lead optimization to introduce substituents at the 6-position of the scaffold, giving the 6-aryl substituted 21b which had improved cellular activity but no oral bioavailability in rat. Property-based optimization at the 6-position and a scaffold change led to the discovery of the imidazo[1,2-b]pyridazine-based 27f, an extremely potent (cellular Mps1 IC50 = 0.70 nM, A549 IC50 = 6.0 nM), selective Mps1 inhibitor over 192 kinases, which could be orally administered and was active in vivo. This 27f demonstrated remarkable antiproliferative activity in the nanomolar range against various tissue cancer cell lines.

Lemierre syndrome is a clinical syndrome that presents with internal jugular thrombophlebitis, septicemia and systemic abscess formations. In general, the condition is preceded by oropharyngeal infections. We report a case of a 73-year-old man with Lemierre syndrome, clivus osteomyelitis and a steroid-responsive mass in the cavernous sinus-suprasellar region. He complained of fever, occipital pain, diplopia and right ptosis. Administration of oral steroids ameliorated the ophthalmic symptoms for a period before he was admitted to our hospital. After admission, the severity of his headache advanced, and his ophthalmic symptoms progressed bilaterally. Brain magnetic resonance imaging showed contrast enhancement of the clivus and revealed a mass lesion contrast-enhancement effect in the cavernous sinus-suprasellar region. Fusobacterium nucleatum was detected by blood culture, and computed tomography revealed multiple bacterial emboli in both lung fields and thrombosis of the left internal jugular vein; thus, he was diagnosed with Lemierre syndrome. After venous administration of antibiotics, his fever and headache markedly improved, but the ophthalmic symptoms did not. We prescribed an oral steroid because the cavernous sinus-suprasellar lesion was probably an inflammatory granuloma caused by a para-infectious mechanism rather than by infection. After the series of treatments, his ophthalmic symptoms improved, and the cavernous sinus-suprasellar region mass lesion decreased. He was eventually discharged in a fully ambulatory state and had no ophthalmic difficulties. We thought that the osteomyelitis of clivus was caused by Lemierre syndrome and its inflammatory processes formed the granuloma in the cavernous sinus-suprasellar region. This was a case of Lemierre syndrome with a rare combination of clivus osteomyelitis and a steroid-responsive tumour in the cavernous sinus-suprasellar region that was successfully treated.

Background: Fatigue is a common nonmotor symptom of Parkinson's disease (PD). Although the causes of fatigue were estimated in the previous reports, fatigue is not fully understood. To determine the frequency of and factors related to fatigue in patients with PD, we carried out clinical assessments in our university hospital., Methods: We used the Japanese version of the Parkinson Fatigue Scale (J-PFS). The J-PFS was administered to 110 patients with PD, and a cutoff point of 3.3 was used for the diagnosis of fatigue. Subsequently, demographic characteristics, clinical features, and medications utilized were evaluated to elucidate the factors related to fatigue. In particular, we focused on the relationship between fatigue and gait disorder assessed via the portable gait rhythmogram., Results: The frequency of fatigue in patients with PD was 52.7%. Univariate analysis revealed that factors significantly associated with fatigue were many motor symptoms and nonmotor symptoms. In addition, multivariate analysis revealed that gait disorder and constipation were independent factors related to fatigue. Furthermore, short-step walking and bradykinesia in gait disorder had especially a relationship with fatigue., Conclusions: More than half of our patients were judged having fatigue. Several factors, including motor and nonmotor symptoms, might be related to fatigue in patients with PD.

Monopolar spindle 1 (Mps1) is an attractive cancer drug target due to the important role that it plays in centrosome duplication, the spindle assembly checkpoint, and the maintenance of chromosomal stability. A design based on JNK inhibitors with an aminopyridine scaffold and subsequent modifications identified diaminopyridine 9 with an IC50 of 37 nM. The X-ray structure of 9 revealed that the Cys604 carbonyl group of the hinge region flips to form a hydrogen bond with the aniline NH group in 9. Further optimization of 9 led to 12 with improved cellular activity, suitable pharmacokinetic profiles, and good in vivo efficacy in the mouse A549 xenograft model. Moreover, 12 displayed excellent selectivity over 95 kinases, indicating the contribution of its unusual flipped-peptide conformation to its selectivity.

Anionic surfactant solutions of sodium dodecylbenzenesulfonate (SDBS) and sodium dodecylsulfate (SDS) separate into two phases through cooling of the solution and the addition of salts. Thiourea (TU) reacts with many metals to form cationic complexes of [M(TU)(n)](m+); some of them extract into the SDBS phase. The separation was examined for lead and tin in commercially purchased solders. More than 95% lead was extracted into the SDBS phase, while tin remained in the aqueous phase. An abnormal decrease in extractability was observed for the [M(TU)(n)](m+) of noble metal complexes at low metal concentrations. The surfactant phases of SDBS and SDS were confirmed as lamellar structures using small angle X-ray scattering (SAXS), and the stability and rigidity of this structure influenced the extractability of the [Au(TU)(n)](3+) complex. The separation of gold(III) and palladium(II) was examined for the SDS system.

A micellar solution of cetylpyridinium chloride (CPC) can separate into two phases due to a temperature change or to the addition of salts. Platinum(II), (IV) and palladium(II) reacted with chloride ions to form stable anionic complexes of PtCl4(2-), PtCl6(2-) and PdCl4(2-), respectively, and were adsorbed onto the CPC gel phase. The CPC phase plays the role of an ion-exchange adsorbent for the anionic complexes. By such a procedure, the precious metals of platinum and palladium could be separated from base metals such as copper, zinc and iron. The kinetic separation was performed by a ligand exchange reaction of the palladium(II) chloro-complex with EDTA at 60 degrees C. The anionic palladium(II)-EDTA complex could not bind the opposite charged CP+ and was desorbed from the CPC phase. In the aqueous phase, the recovery of palladium(II) by the double-desorption was 101.1 +/- 1.2%. The platinum(II) and (IV) chloro-complexes were stable for at least 30 min and remained in the CPC phase.

A micellar solution of sodium dodecylsulfate (SDS) exhibits the property of being separated into two phases due to a temperature change or the addition of salts. The ammine-complexes of copper(II), nickel(II) and palladium(II) reacted with the dodecylsulfate anion to form the corresponding ion-pair, and were extracted into the SDS gel phase. The SDS plays the roles of a pairing-ion for the ammine-complexes and of an extraction medium. The ion-pair extraction mechanism was investigated; the extractability of metals was given by the function of the solubility products of the ion-pairs. This method was applied to the mutual separation of Ni(II)/Cu(II) and Pd(II)/Pt(II). The driving force for the extraction was an electrostatic interaction between the cationic complex and the surfactant anion. The use of the SDS gel as ion-exchanger is also expected.

The extraction and separation of copper(II), zinc(II), cobalt(II), and cadmium(II) were investigated. Both copper(II) and zinc(II) formed ammine-complexes, while cadmium(II) and cobalt(II) formed hydroxide precipitates in an ammonia medium. By the addition of sodium dodecylsulfate (SDS), a copper(II) complex formed an ion-pair (copper-ammine-DS), which was extracted into the SDS phase. However, a zinc(II) complex did not form an ion-pair, and was soluble in water. Copper(II) ion was recovered by stripping (back-extraction) after the addition of hydrochloric acid. This method was applied to the separation of copper(II) in a brass alloy.

Akt is known to be activated in the rheumatoid synovial tissues. We examined here functional role of Akt during tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-mediated apoptosis in rheumatoid synovial cells. Rheumatoid synovial cells in vitro were rapidly committed to apoptosis in response to TRAIL in mitochondria-dependent manner whereas Akt and extracellular signal-regulated kinase (ERK) were also phosphorylated. TRAIL-mediated apoptosis in synovial cells was significantly increased through inactivation of Akt by LY294002, however, that process was not so changed by adding ERK inhibitor, PD98059. Platelet-derived growth factor (PDGF) clearly phosphorylated both Akt and ERK in synovial cells, and PDGF pretreatment markedly suppressed TRAIL-mediated synovial cell apoptosis. The use of not PD98059 but LY294002 abrogated PDGF-mediated inhibitory effect toward TRAIL-induced apoptosis in synovial cells. The above protective effect of Akt was confirmed by the use of short interfering RNA (siRNA)-directed inhibition of Akt. Our data suggest that Akt is an endogenous inhibitor during TRAIL-mediated synovial cell apoptotic pathway, which may explain that synovial cells in situ of the rheumatoid synovial tissues are resistant toward apoptotic cell death in spite of death receptor expression.

Cbfa1 is an essential transcription factor for bone formation, but its transcripts have also been detected in thymus and testis. To elucidate the expression of Cbfa1 in testis, we isolated Cbfa1 cDNA from mouse testis. First we examined the length of the transcripts expressed in mouse testis by Northern hybridization. Using a cDNA probe that can detect both Type-I and Type-II Cbfa1 (Type-I: originally reported as Pebp2alphaA by Ogawa et al.; Type-II: originally reported as til-1 by Stewart et al.), 1.8-kb transcripts were detected in testis, and larger transcripts (6.3 kb and 7.4 kb) in T-cell lines, thymus and bone. Using a Type-II specific probe (bone isoform-specific probe), surprisingly, 1.8-kb testis transcript(s) were detected as well as those (6.3 kb) found in bone. In addition, the transcription start site in mouse testis coincides with one of three start sites identified in bone. These results suggest that the testis transcript is generated from the same promoter of Type-II Cbfa1, which is thought to be active only in osteoblasts and some chondrocytes. Next, to define the difference in length of transcripts, we isolated the Cbfa1 cDNA from mouse testis. Sequence analysis of the cDNA showed that alternative splicing around exon 2 and poly-adenylation within exon 8 occurred in the testis isoform, which produce the smaller transcripts. These data revealed that testis Cbfa1 mRNA is transcribed from same promoter used in bones, but the post-transcriptional regulation is different between testis and skeletal tissues.

Cbfa1 is an essential transcription factor for bone formation, and as such little is known about the region responsible for the transcriptional regulation of this gene. Here we report the determination of the transcription start sites, isolation and partial characterization of distal promoter region of this gene. Three transcription start sites were identified by the 5'-Cap site method, recently invented for rapid examination of the 5'-end of genes of interest. A reporter construct containing 1.8 kb of 5' of transcription start sites had approximately 25-fold more luciferase activity than the promoter-less vector in osteoblastic cell lines. Deletion analysis of the reporter construct demonstrated that the minimal region to express promoter activity lies between bp -168 and -99, taking the most downstream transcription start site as +1. By Northern blot analysis, mRNA expression from the distal promoter was detected in the differentiated osteoblastic cell lines, UMR-106, ROS17/2.8 and MC3T3-E1, but not in cell lines of immature phenotype or originated from other organs. Luciferase activity was strongest in UMR-106 and ROS17/2.8, and weakest in COS-1 and HepG2, which are cell lines originating from other organs, corresponding to the level of mRNA expression. These results demonstrated that the distal promoter region examined here is important for tissue- and cell-type-specific gene expression of Cbfa1.

Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. We investigated functional differences among three isoforms of Cbfa1: Type I (originally reported as Pebp2alphaA by Ogawa et al. (Ogawa, E., Maruyama, M., Kagoshima, H., Inuzuka, M., Lu, J., Satake, M., Shigesada, K., and Ito, Y. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 6859-6863), Type II (originally reported as til-1 by Stewart et al. (Stewart, M., Terry, A., Hu, M., O'Hara, M., Blyth, K., Baxter, E., Cameron, E., Onions, D. E., and Neil, J. C. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 8646-8651), and Type III (originally reported as Osf2/Cbfa1 by Ducy et al. (Ducy, P., Zhang, R., Geoffroy, V., Ridall, A. L., and Karsenty, G. (1997) Cell 89, 747-754). A reverse transcriptase-polymerase chain reaction analysis demonstrated that these isoforms were expressed in adult mouse bones. The transient transfection of Type I or Type II Cbfa1 in a mouse fibroblastic cell line, C3H10T1/2, induced the expression of alkaline phosphatase (ALP) activity. This induction was synergistically enhanced by the co-introduction of Xenopus BMP-4 cDNA. In contrast, the transient transfection of Type III cDNA induced no ALP activity. In C3H10T1/2 cells stably transfected with each isoform of Cbfa1, the gene expression of ALP was also strongly induced in cells transfected with Type I and Type II Cbfa1 but not in cells with Type III Cbfa1. Osteocalcin, osteopontin,and type I collagen gene expressions were induced or up-regulated in all of the cells stably transfected with each isoform of Cbfa1, and Type II transfected cells exhibited the highest expression level of osteocalcin gene. A luciferase reporter gene assay using a 6XOSE2-SV40 promoter (6 tandem binding elements for Cbfa1 ligated in front of the SV40 promoter sequence), a mouse osteocalcin promoter, and a mouse osteopontin promoter revealed the differences in the transcriptional induction of target genes by each Cbfa1 isoform with or without its beta-subunit. These results suggest that all three of the Cbfa1 isoforms used in the present study are involved in the stimulatory action of osteoblast differentiation, but they exert different functions in the process of osteoblast differentiation.

We investigated the relationship between the body mass index and the prevalence of coronary risk factors, hypertension, hyperlipidemia, glucose intolerance and hyperuricemia, in middle-aged male workers. We found a positive correlation between the body mass index and the prevalence rate of these coronary risk factors. Mean values for serum total cholesterol, triglyceride, LDL-cholesterol and HbA1c increased with the increase in the body mass index, but decreased in HDL-cholesterol.

We cloned the mouse homolog of FGF10, which was recently reported as a new member of the FGF family. The predicted molecular mass of this molecule is 23.6 kDa, and both nucleotide and amino acid sequences show high degrees of similarity with those of the rat. Examination of mouse FGF10 mRNA expression in various tissues and developmental stages by Northern hybridization revealed tissue- and developmental stage-specific expression of the gene. Similarly to the rat counterpart, mouse FGF10 mRNA (4.5 kb) was expressed relatively abundantly in embryos and the lung, and at much lower levels in brain and heart. In addition, a shorter transcript (1.3 kb) is expressed only in testis. Considering the high similarity in primary structure between FGF10 and FGF7 (known as keratinocyte growth factor; KGF), we also examined the gene expression of FGF10 during wound healing using a mouse model. FGF10 mRNA was highly induced 1 day after injury and decreased rapidly by 3 days. This suggests that FGF10 is a primary factor in the process of wound healing similarly to other growth factors such as TGF alpha and FGF7.

We isolated the cDNA encoding a novel member of the human fibroblast growth factor (FGF) family from the lung. The cDNA encodes a protein of 208 amino acids with high sequence homology (95.6%) to rat FGF-10, indicating that the protein is human FGF-10. Human FGF-10 as well as rat FGF-10 has a hydrophobic amino terminus ( approximately 40 amino acids), which may serve as a signal sequence. The apparent evolutionary relationships of human FGFs indicate that FGF-10 is closest to FGF-7. Chromosomal localization of the human FGF-10 gene was examined by in situ hybridization. The gene was found to map to the 5p12-p13 region. Human FGF-10 (amino acids 40 to 208 with a methionine residue at the amino terminus) was produced in Escherichia coli and purified from the cell lysate. Recombinant human FGF-10 (approximately 19 kDa) showed mitogenic activity for fetal rat keratinizing epidermal cells, but essentially no activity for NIH/3T3 cells, fibroblasts. The specificity of mitogenic activity of FGF-10 is similar to that of FGF-7 but distinct from that of bFGF. In structure and biological activity, FGF-10 is similar to FGF-7.

We isolated the cDNA encoding a novel member of the fibroblast growth factor (FGF) family from rat embryos by homology-based polymerase chain reaction. The FGF-related cDNA encodes a protein of 215 amino acids (approximately 24 kDa), which has a conserved approximately 120-amino acid core with approximately 30-60% amino acid sequence identity with the FGF family. This protein with a hydrophobic amino terminus appears to be a secreted protein. The cDNA was translated in a coupled in vitro transcription-translation system. The molecular mass of the translation product was observed to be approximately 26 kDa. The expression of the FGF-related mRNA in the rat embryo and adult tissues was determined by Northern analysis and in situ hybridization. The mRNA was expressed in several discrete regions of the embryo. In adult tissues, the mRNA was preferentially expressed in the lung. The expression profile of the FGF-related mRNA was different from those of other FGF family mRNAs. As this protein is the 10th documented protein related to FGFs, we tentatively term this protein FGF-10.

We examined the localization of fibroblast growth factor-9 (FGF-9) mRNA in the rat brain by in situ hybridization. FGF-9 mRNA was moderately or weakly expressed in widespread regions including the olfactory bulb, caudate putamen, cerebral cortex, hippocampus, thalamus, hypothalamus, midbrain, brainstem and cerebellum. However, FGF-9 mRNA was also strongly expressed in several specific nuclei including the red nucleus and oculomotor nucleus in the midbrain, the vestibular nucleus and facial nucleus in the brainstem and the medial cerebellar nucleus, interposed cerebellar nucleus and lateral cerebellar nucleus in the cerebellum. The cellular localization of FGF-9 mRNA indicated that the mRNA in the rat brain was expressed preferentially in neurons, although FGF-9 was originally isolated from human glioma cells. The localization profile of FGF-9 mRNA is different from those of aFGF, bFGF and FGF-5 mRNAs reported previously. The present findings indicate that FGF-9 has a unique role in the brain.

The fibroblast growth factor (FGF) receptor family consists of four members, FGFR-1, FGFR-2, FGFR-3 and FGFR-4, that are closely related receptor tyrosine kinases. We examined the expression of rat FGFR-4 mRNA in the brain by in situ hybridization and compared it with that of the mRNAs for other FGF receptors. In contrast with FGFR-1, FGFR-2 and FGFR-3 mRNAs which are expressed widely in the brain, the FGFR-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus neurons. The present finding indicates that FGFR-4 has a function specific to the medial habenular nucleus.