Loss of DIAPH3, a Formin Family Protein, Leads to Cytokinetic Failure Only under High Temperature Conditions in Mouse FM3A Cells.

Cell division is essential for the maintenance of life and involves chromosome segregation and subsequent cytokinesis. The processes are tightly regulated at both the spatial and temporal level by various genes, and failures in this regulation are associated with oncogenesis. Here, we investigated the gene responsible for defects in cell division by using murine temperature-sensitive (ts) mutant strains, tsFT101 and tsFT50 cells. The ts mutants normally grow in a low temperature environment (32 °C) but fail to divide in a high temperature environment (39 °C). Exome sequencing and over-expression analyses identified Diaph3, a member of the formin family, as the cause of the temperature sensitivity observed in tsFT101 and tsFT50 cells. Interestingly, Diaph3 knockout cells showed abnormality in cytokinesis at 39 °C, and the phenotype was rescued by re-expression of Diaph3 WT, but not Diaph1 and Diaph2, other members of the formin family. Furthermore, Diaph3 knockout cells cultured at 39 °C showed a significant increase in the level of acetylated α-tubulin, an index of stabilized microtubules, and the level was reduced by Diaph3 expression. These results suggest that Diaph3 is required for cytokinesis only under high temperature conditions. Therefore, our study provides a new insight into the mechanisms by which regulatory factors of cell division function in a temperature-dependent manner.

Clinical Subtypes in Children with Attention-Deficit Hyperactivity Disorder According to Their Child Behavior Checklist Profile.

This study sought to identify subgroups of attention-deficit hyperactivity disorder (ADHD) defined by specific patterns of emotional and behavioral symptoms according to the parent-rated Child Behavior Checklist (CBCL). Our clinical sample comprised 314 children (aged 4 to 15 years) diagnosed with ADHD according to the DSM-5. In addition, comorbid psychiatric disorders, general functioning, and medication status were assessed. Cluster analysis was performed on the CBCL syndrome subscales and yielded a solution with four distinct subgroups. The "High internalizing/externalizing" group displayed an overlap between internalizing and externalizing problems in the CBCL profile. In addition, the "High internalizing/externalizing" group revealed a high rate of comorbid autism spectrum disorder and elevated autistic traits. The "Inattention and internalizing" group revealed a high rate of the predominantly inattentive presentation according to ADHD specifier from the DSM-5. The "Aggression and externalizing" group revealed a high rate of comorbid oppositional defiant disorder and conduct disorder. The "Less psychopathology" group scored low on all syndrome scales. Children with ADHD were subdivided into four distinct subgroups characterized by psychopathological patterns, with and without internalizing and externalizing problems. The overlap between internalizing and externalizing problems may be mediated with emotional dysregulation and associated neurobiological bases.

Distinct chemotherapy-associated anti-cancer immunity by myeloid cells inhibition in murine pancreatic cancer models.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy associated with an extremely poor prognosis. Chemotherapy, such as gemcitabine (GEM), is the only treatment for PDAC patients who are not suitable for radical surgical treatment; however, its anti-tumor efficacy is limited. In this study, we investigated the host immune system response in murine PDAC models undergoing GEM treatment. We found that PDAC tumor tissues were infiltrated with a substantial number of Gr-1+ myeloid cells and had relatively small numbers of CD4+ and CD8+ cells. In addition, there were increased numbers of myeloid cells expressing CD11b+ and Gr-1+ in peripheral blood. When mice with PDAC tumors in the intraperitoneal cavity or liver were treated with GEM, numbers of myeloid cells in tumor tissues and in peripheral blood decreased. In contrast, numbers of CD4+ or CD8+ cells increased. In peripheral blood, the numbers of CD8+ cells expressing interferon-gamma (IFN-γ) were higher in GEM-treated mice than in untreated mice. In addition, GEM treatment in combination with myeloid cell depletion further prolonged the survival of PDAC mice. The gene expression profile of peripheral blood in myeloid cell-depleted PDAC mice treated with GEM showed biological processes related to anti-cancer immunity, such as natural killer cell-mediated cytotoxicity, type I IFN signaling, and co-stimulatory signaling for T cell activation. Thus, in PDAC murine models, GEM treatment was associated with an immune response consistent with an anti-cancer effect, and depletion of myeloid-lineage cells played an important role in enhancing anti-cancer immunity associated with GEM treatment.

Adipose tissue-derived stem cells prevent fibrosis in murine steatohepatitis by suppressing IL-17-mediated inflammation.

BACKGROUND AND AIM: The pathological features of non-alcoholic steatohepatitis (NASH) have not been determined, so fundamental treatment has not been established. Adipose-tissue-derived stromal/stem cells (ADSCs) are beneficial for repair/regenerative therapy of impaired organs because of their immuno-modulatory capability. In this study, we assessed how liver damage progresses during the early development phase of the murine NASH model and investigated whether ADSCs are preventatively efficacious against the fibrosis progression of NASH. METHODS: C57BL/6J mice were fed with atherogenic high fat or high-fat diet 60 developing into NASH or simple steatosis. Their hepatic inflammatory cells (HICs) were analyzed by cDNA microarray. NASH mice were treated with ADSCs injected into spleen when hepatic inflammation was initially observed, and liver samples were analyzed. The effect of ADSCs on the mice hepatic stellate cell (HSC) line stimulated by recombinant IL-17 and HICs from NASH mice was analyzed. RESULTS: The gene expression features of HICs implicated as humoral cytokine mediators of lymphoid cells during NASH development, compared with a simple steatosis model. One of the featured cytokines was IL-17. The development of hepatic fibrosis was alleviated when NASH mice were treated with ADSCs as well as treated with anti-IL-17 antibody, and the frequency of IL-17-secreting HICs decreased. NASH-HICs enhanced proliferation of HSCs, in which proliferation was sensitive to IL-17 stimulation. The stimulatory effect of NASH-HICs on the activation of HSCs was attenuated by co-culture with ADSCs. CONCLUSION: ADSCs treatment prevented progression of NASH fibrosis by suppressing IL-17-mediated inflammation, which was associated with HSCs activation.

Foetal abuse.

Pregnancy and motherhood are often presented as natural and unproblematic for women. The fact that there are some women who engage in violent behaviour towards their unborn child suggests that motherhood is not as unproblematic as we are led to believe. This paper presents six previously unpublished cases of foetal abuse that is physical assaults on the foetus by the mothers themselves, and examines how the prevailing myth of the good mother might be further endangering mothers and their unborn children. So far, the research suggests there are some common, possibly co-occurring, features that might be an antecedent to foetal abuse: unplanned pregnancies, prior mental health issues in the mother, trauma, pregnancy denial up to 20 weeks or until birth, and ideation of harm correlated to in utero movements.

The CD45+ fraction in murine adipose tissue derived stromal cells harbors immune-inhibitory inflammatory cells.

Stromal cells in adipose tissue are useful for repair/regenerative therapy as they harbor a substantial number of mesenchymal stem cells; therefore, freshly isolated autologous uncultured adipose tissue derived stromal cells (u-ADSCs) are useful for regenerative therapy, and obviate the need for mesenchymal stem cells. We evaluated the therapeutic effect of murine u-ADSCs and sorted subsets of u-ADSCs in a concanavalin A (ConA) induced murine model of hepatitis, as well as their characteristics. We found that 10-20% of u-ADSCs expressed the CD45 leukocyte-related antigen. CD68, which is a marker of macrophages (MΦs), was expressed by 50% of CD45+ u-ADSCs. About 90% of CD68+ CD45+ cells expressed CD206 antigen, which is a marker of inhibitory M2-type MΦs. Genes related to M2-type MUs were especially more highly expressed by CD45+ CD206+ u-ADSCs than by CD45- u-ADSCs. CD45+ u-ADSCs inhibited the expression of cytokines/chemokines and suppressed the proliferation of splenocytes stimulated with ConA. We observed that not only whole u-ADSCs, but also the CD45+ subset of u-ADSCs ameliorated the ConA-induced hepatitis in mice. In conclusion, we show that freshly isolated murine u-ADSCs were effective against acute hepatitis, and CD45+ u-ADSCs acting phenotypically and functionally like M2-type MΦs, contributed to the repair of liver tissue undergoing inflammation.

Clinical features of cystatin A expression in patients with pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is the most lethal malignancy known, with an extremely poor prognosis due to the lack of an efficient diagnostic scheme and no radical treatment option, except surgery. Therefore, understanding the pathophysiology of, and finding a novel biomarker to detect, PDAC should be prioritized. We observed an increase in mRNA expression of the cysteine protease inhibitor cystatin A (CSTA) in CD4+ T cells in peripheral blood cells of nine patients with PDAC, compared with the expression in seven healthy volunteers. Moreover, we confirmed significantly higher CSTA mRNA expression in a larger cohort of 41 patients with PDAC compared with that in 20 healthy volunteers. Correspondingly, the serum CSTA concentrations in 36 patients with PDAC were higher than those in 37 healthy volunteers, and this increase was correlated with PDAC clinical stage. Furthermore, the expression of CSTA and cathepsin B, which is a lysosomal cysteine protease inhibited by CSTA, was observed in tumor tissues and tumor-infiltrating immune cells in 20 surgically resected PDAC tissues by immunohistochemical staining. Expression of CSTA was detected in some tumor tissues and many tumor-infiltrating immune cells. Cathepsin B expression was also observed in most tumor tissues and tumor-infiltrating immune cells. In conclusion, CSTA and its substrate cathepsin B are involved in PDAC-related inflammation. The increment of CSTA expression in peripheral blood of patients with PDAC may have a potential role as a PDAC immunopathologic biomarker.

SPONTANEOUS LIPOPROTEIN GLOMERULOPATHY-LIKE NEPHROPATHY IN A SQUIRREL (SCIURUS VULGARIS).

Lipoprotein glomerulopathy (LPG) is a rare human glomerular disease caused by abnormal lipid metabolism. Naturally occurring LPG has not been reported in animals. We describe the histopathological characterization of spontaneous LPG-like nephropathy in a captive squirrel ( Sciurus vulgaris ). Macroscopically, swollen glomeruli were distinctively identified as fine white granules in the renal cortex. Histologically, most glomeruli were markedly enlarged with distended capillaries containing faintly eosinophilic and amorphous materials. The amorphous material was negative using the periodic acid-Schiff reaction, periodic acid-methenamine silver stain, or Masson's trichrome stain. Sudan III staining revealed lipid in the materials, and immunohistochemistry demonstrated that the material additionally contained apolipoprotein E. Electron microscopy showed numerous lipid granules and vacuoles of various sizes in the capillary lumina associated with foot process effacement of podocytes. These pathological characteristics bear some resemblance to those of human LPG.

Adipose tissue derived stromal stem cell therapy in murine ConA-derived hepatitis is dependent on myeloid-lineage and CD4+ T-cell suppression.

Mesenchymal stromal stem cells (MSCs) are an attractive therapeutic model for regenerative medicine due to their pluripotency. MSCs are used as a treatment for several inflammatory diseases, including hepatitis. However, the detailed immunopathological impact of MSC treatment on liver disease, particularly for adipose tissue derived stromal stem cells (ADSCs), has not been described. Here, we investigated the immuno-modulatory effect of ADSCs on hepatitis using an acute ConA C57BL/6 murine hepatitis model. i.v. administration of ADSCs simultaneously or 3 h post injection prevented and treated ConA-induced hepatitis. Immunohistochemical analysis revealed higher numbers of CD11b(+), Gr-1(+), and F4/80(+) cells in the liver of ConA-induced hepatitis mice was ameliorated after the administration of ADSCs. Hepatic expression of genes affected by ADSC administration indicated tissue regeneration-related biological processes, affecting myeloid-lineage immune-mediating Gr-1(+) and CD11b(+) cells. Pathway analysis of the genes expressed in ADSC-treated hepatic inflammatory cells revealed the possible involvement of T cells and macrophages. TNF-α and IFN-γ expression was downregulated in hepatic CD4(+) T cells isolated from hepatitis livers co-cultured with ADSCs. Thus, the immunosuppressive effect of ADSCs in a C57BL/6 murine ConA hepatitis model was dependent primarily on the suppression of myeloid-lineage cells and, in part, of CD4(+) T cells.

Adipose tissue-derived stem cells as a regenerative therapy for a mouse steatohepatitis-induced cirrhosis model.

UNLABELLED: Cirrhosis is a chronic liver disease that impairs hepatic function and causes advanced fibrosis. Mesenchymal stem cells have gained recent popularity as a regenerative therapy since they possess immunomodulatory functions. We found that injected adipose tissue-derived stem cells (ADSCs) reside in the liver. Injection of ADSCs also restores albumin expression in hepatic parenchymal cells and ameliorates fibrosis in a nonalcoholic steatohepatitis model of cirrhosis in mice. Gene expression analysis of the liver identifies up- and down-regulation of genes, indicating regeneration/repair and anti-inflammatory processes following ADSC injection. ADSC treatment also decreases the number of intrahepatic infiltrating CD11b(+) and Gr-1(+) cells and reduces the ratio of CD8(+) /CD4(+) cells in hepatic inflammatory cells. This is consistent with down-regulation of genes in hepatic inflammatory cells related to antigen presentation and helper T-cell activation. CONCLUSION: These results suggest that ADSC therapy is beneficial in cirrhosis, as it can repair and restore the function of the impaired liver.

Colloid carcinoma of the uterine cervix: a case report with respect to immunohistochemical analyses.

Colloid carcinoma, characterized by the presence of a large amount of extracellular mucin that results in the formation of mucous lakes with a relative paucity of neoplastic glandular cells within them, is extremely rare in the uterine cervix. Herein, we report an additional case of colloid carcinoma of the cervix and discuss the immunohistochemical characteristics and histogenesis of this extremely rare tumor. A 47-year-old Japanese female without any history of carcinomas was found to have a bulky mass in the cervix. Biopsy from the cervix revealed adenocarcinoma; subsequently, total hysterectomy was performed. Histopathologic study demonstrated that columnar or cuboidal neoplastic glandular cells forming cribriform or tubular structures floated within the mucous lakes involving almost the entire layer of the cervical wall. Adenocarcinoma in situ (AIS) component was also observed. Immunohistochemically, tumor cells of the colloid carcinoma were positive for cytokeratin 7, MUC5AC, MUC6, and p16 (diffuse), but negative for cytokeratin 20, MUC2, and cdx-2. In addition, human papillomavirus 16 was detected in both colloid carcinoma and AIS components. This is the first reported case of endocervical type colloid carcinoma, and the second documented case of cervical colloid carcinoma with immunohistochemical analyses of mucin. The present case had an endocervical type AIS component, which suggests that AIS may be a precursor lesion of colloid carcinoma. Moreover, this case clearly demonstrates that the occurrence of cervical colloid carcinoma correlates with high-risk human papillomavirus.

Impact of a fixed price system on the supply of institutional long-term care: a comparative study of Japanese and German metropolitan areas.

BACKGROUND: The need for institutional long-term care is increasing as the population ages and the pool of informal care givers declines. Care services are often limited when funding is controlled publicly. Fees for Japanese institutional care are publicly fixed and supply is short, particularly in expensive metropolitan areas. Those insured by universal long-term care insurance (LTCI) are faced with geographically inequitable access. The aim of this study was to examine the impact of a fixed price system on the supply of institutional care in terms of equity. METHODS: The data were derived from official statistics sources in both Japan and Germany, and a self-administered questionnaire was used in Japan in 2011. Cross-sectional multiple regression analyses were used to examine factors affecting bed supply of institutional/residential care in fixed price and free prices systems in Tokyo (Japan), and an individually-bargained price system in North Rhine-Westphalia (Germany). Variables relating to costs and needs were used to test hypotheses of cost-dependency and need-orientation of bed supply in each price system. Analyses were conducted using data both before and after the introduction of LTCI, and the results of each system were qualitatively compared. RESULTS: Total supply of institutional care in Tokyo under fixed pricing was found to be cost-dependent regarding capital costs and scale economies, and negatively related to need. These relationships have however weakened in recent years, possibly caused by political interventions under LTCI. Supply of residential care in Tokyo under free pricing was need-oriented and cost-dependent only regarding scale economies. Supply in North Rhine-Westphalia under individually bargained pricing was cost-independent and not negatively related to need. CONCLUSIONS: Findings suggest that publicly funded fixed prices have a negative impact on geographically equitable supply of institutional care. The contrasting results of the non-fixed-price systems for Japanese residential care and German institutional care provide further theoretical supports for this and indicate possible solutions against inequitable supply.

[Impact of psychotropic drugs on breast-fed Infants: international perspective].

The benefit of breast-feeding has been established, and it has recently been strongly encouraged internationally. However, new mothers have more risks of developing psychiatric symptoms than in any other period of their life, and some of them require psychopharmacotherapy. There are some clinical dilemmas regarding the benefit of such therapy and possible adverse effects on infants when mothers want to breast-feed their infants. Most drugs are transferred to breast-milk and, eventually, low levels of maternal daily doses can be taken by breast-fed infants. Having considered this issue and based on several studies, the authors of studies report that breast-feeding can be carried out by mothers who take psychotropic drugs. More research data accumulation, including individual case observations and follow-ups, is necessary to guarantee the safety of breast-feeding by a mother on medication; however, in the current clinical setting, decision-making is essential to plan treatment. Such mothers are allowed to breast-feed infants who can be monitored by pediatricians. The critical matter is that clinicians should not hesitate to prescribe optimal doses to mothers for effective treatment.

[Education and training system in child and adolescent psychiatry with view to carry-over towards adulthood psychiatry].

AIM: The aim of our study was to develop and establish a training system in child and adolescent psychiatry in Japan, especially in light of the continuity and integration with general psychiatry. METHODS: The Japanese authors surveyed the situation of training in child and adolescent psychiatry (CAP training) through the literature and conducted collaborative and consensus meetings with the authors in the UK, with its long history of training system development. A comparison was made between Japan and CAP training in the UK and other countries. RESULTS: A recent survey of psychiatric education in Japan clarified the current situation and called attention to parts of the education and training system where development is required. A systematic curriculum has not been established. For the elimination of disparities of competencies and skills among multidisciplinary staff, the CAP training curriculum needs to be comprehensive and fundamental, as shown in the Competency-based Curriculum for Specialists developed by the Royal College of Psychiatrists. CONCLUSIONS: CAP training and general psychiatry must be integrated into a comprehensive curriculum in the postgraduate education and training system, with an adequate time and place for training and supervision.

Natto (fermented soybean) extract extends the adult lifespan of Caenorhabditis elegans.

We investigated the effects of a water extract of natto on the aging of the nematode Caenorhabditis elegans. The water extract significantly prolonged the adult lifespan of the wild-type worms and rendered them resistant to oxidative and thermal stress. In addition, treatment with natto extract significantly delayed the accumulation of lipofuscin, a characteristic of aging cells. Our findings suggest that components of natto have a beneficial anti-aging effect in vivo.

Association of changes in the gene expression profile of blood cells with the local tumor inflammatory response in a murine tumor model.

Cancer tissue is frequently associated with the host inflammatory response, which involves blood cells. Using DNA microarrays, we examined the gene expression profiles of blood and tumors in a murine subcutaneous hepatocellular carcinoma model, in which tumors develop during the initial 10 days and then diminish and disappear by day 25 after implantation. Immunohistochemical and gene expression analysis indicated that tumor tissues were associated with an active immune response, particularly the CD4+ T cell-mediated immune response, on day 10. The genes commonly up-regulated in blood and the fraction enriched with tumor-associated inflammatory cells on day 10 also suggested the involvement of CD4+ T cells. Unsupervised hierarchical clustering analysis of gene expression of peripheral blood cells on days 0, 10, 15, 20, and 25 indicated two major clusters: the tumor-existence cluster on days 10, 15, and 20, and the tumor-free cluster on days 0 and 25. Additionally, sub-clusters were detected on each day. These results suggest that the gene expression profile of whole blood cells is affected by the local tumor condition, and is associated with the local host immune response. Its analysis will facilitate exploration of the underlying important features of the host immune response to tumors.

A case of subacute thrombosis associated with clopidogrel resistance after implantation of a zotarolimus-eluting stent.

A 73-year-old woman was admitted to our hospital with anterior acute myocardial infarction due to subacute thrombosis after coronary stenting with a zotarolimus-eluting stent (ZES), which is a newly developed drug-eluting stent that has been widely used since May 2009 in Japan. Five days before, she underwent implantation with a ZES in the left anterior descending artery due to stable angina pectoris. After stenting, the intravascular ultrasonography showed no malapposition from the proximal to the distal edge of the stent. She received aspirin 100 mg/day and clopidogrel 75 mg/day from 2 weeks before the stent was implanted. When we investigated the single nucleotide polymorphisms of CYP2C19 in this patient, both CYP2C19*2 and CYP2C19*3 were detected, and she was classified as a poor metabolizer. This report is the first to describe subacute stent thrombosis following the implantation of a newly developed ZES in a Japanese patient, which may be related to clopidogrel resistance.

A Japanese version of Mother-to-Infant Bonding Scale: factor structure, longitudinal changes and links with maternal mood during the early postnatal period in Japanese mothers.

The objectives of this study were (1) to develop a Japanese version of Mother-to- Infant Bonding Scale Japanese version (MIBS-J) based on Kumar's Mother Infant Bonding Questionnaire that could be used to screen the general population for problems in the mother's feelings towards her new baby and to validate it for clinical use and (2) to examine the factor structure of the items and create subscales of the questionnaire for the Japanese version. The MIBS-J is a simple self-report questionnaire designed to detect the problems in a mother's feelings towards her newborn baby. Participants (n = 554) were recruited at an outpatient clinic of a maternity hospital in a community after 30-weeks gestation. MIBS-J and the Edinburgh Postnatal Depression Scale (EPDS) were administered on the fifth day at the maternity ward and mailed at 1 and 4 months postnatally. Exploratory factor analysis and confirmatory factor analysis demonstrated a two-factor structure out of eight items: lack of affection (LA) and anger/rejection (AR). Chronbach's α coefficients were 0.71 and 0.57, respectively. The LA and AR scores had strong correlations across postnatal times. The mothers with higher (worse) AR scores on the MIBS-J at any of the three periods had higher scores on the EPDS. MIBS-J demonstrated acceptable reliability and reasonable construct validity in this Japanese sample.

Phylogenetic relationships of three species within the family Heligmonellidae (Nematoda; Heligmosomoidea) from Japanese rodents and a lagomorph based on the sequences of ribosomal DNA internal transcribed spacers, ITS-1 and ITS-2.

Nematodes of the family Heligmonellidae (Heligmosomoidea; Trichostrongylina) reside in the digestive tracts of rodents and lagomorphs. Although this family contains large numbers of genera and species, genetic information on the Heligmonellidae is very limited. We collected and isolated adult worms of three species in Japan that belong to the family Heligmonellidae, namely Heligmonoides speciosus (Konno, 1963) Durette-Desset, 1970 (Hs) from Apodemus argenteus, Orientostrongylus ezoensis Tada, 1975 (Oe) from Rattus norvegicus and Lagostrongylus leporis (Schulz, 1931) (Ll) from Pentalagus furnessi, and sequenced the entire internal transcribed spacer regions, ITS-1 and ITS-2 of ribosomal DNA. ITS-1 of Hs, Oe and Ll was 426, 468 and 449 bp in length, and had a G+C content of about 41, 41 and 37 %, respectively. ITS-2 of Hs, Oe and Ll was 297, 319 and 276 bp in length and had a G+C content of about 38, 40 and 28%, respectively. The data of Hs, Oe and Ll were compared with those of two other known species within the family Heligmonellidae, Calorinensis minutus (Dujardin, 1845) (Cm) and Nippostrogylus brasiliensis (Travassos, 1914) (Nb), and with those of two species of Heligmosomidae (Heligmosomoidea), Heligmosomoides polygyrus bakeri and Ohbayashinema erbaevae. Phylogenetic analysis placed Hs, Oe and Ll in the same clade with Cm and Nb, forming a Heligmonellidae branch in both ITS-1 and ITS-2, separate from the Heligmosomoidea branch. These results demonstrated that the ITS-1 and ITS-2 sequences are useful for differentiating the Heligmonellidae nematode species. This study is the first to describe the ITS-1 and ITS-2 sequences of Hs, Oe and Ll.

Podocan-like protein: a novel small leucine-rich repeat matrix protein in bone.

Recently, significant attention has been drawn to the biology of small leucine-rich repeat proteoglycans (SLRPs) due to their multiple functionalities in various cell types and tissues. Here, we characterize a novel SLRP member, "Podocan-like (Podnl) protein" identified by a bioinformatics approach. The Podnl protein has a signal peptide, a unique cysteine-rich N-terminal cluster, 21 leucine-rich repeat (LRR) motifs, and one putative N-glycosylation site. This protein is structurally similar to podocan in SLRPs. The gene was highly expressed in mineralized tissues and in osteoblastic cells and the high expression level was observed at and after matrix mineralization in vitro. Podnl was enriched in newly formed bones based on immunohistochemical analysis. When Podnl was transfected into osteoblastic cells, the protein with N-glycosylation was detected mainly in the cultured medium, indicating that Podnl is a secreted N-glycosylated protein. The endogenous Podnl protein was also present in bone matrix. These data provide a new insight into our understanding of the emerging SLRP functions in bone formation.