Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Fusanori NISHIMURA Last modified date:2019.06.20

Professor / Division of Oral Rehabilitation / Department of Dental Science / Faculty of Dental Science


Papers
1. E. Kakuta, Y. Nomura, T. Morozumi, T. Nakagawa, T. Nakamura, K. Noguchi, A. Yoshimura, Y. Hara, O. Fujise, Fusanori Nishimura, T. Kono, M. Umeda, M. Fukuda, T. Noguchi, N. Yoshinari, C. Fukaya, S. Sekino, Y. Numabe, N. Sugano, K. Ito, H. Kobayashi, Y. Izumi, H. Takai, Y. Ogata, S. Takano, M. Minabe, A. Makino-Oi, A. Saito, Y. Abe, S. Sato, F. Suzuki, K. Takahashi, T. Sugaya, M. Kawanami, N. Hanada, S. Takashiba, H. Yoshie, Assessing the progression of chronic periodontitis using subgingival pathogen levels
A 24-month prospective multicenter cohort study, BMC Oral Health, 10.1186/s12903-017-0337-x, 17, 1, 2017.01, Background: The diagnosis of the progression of periodontitis presently depends on the use of clinical symptoms (such as attachment loss) and radiographic imaging. The aim of the multicenter study described here was to evaluate the diagnostic use of the bacterial content of subgingival plaque recovered from the deepest pockets in assessing disease progression in chronic periodontitis patients. Methods: This study consisted of a 24-month investigation of a total of 163 patients with chronic periodontitis who received trimonthly follow-up care. Subgingival plaque from the deepest pockets was recovered and assessed for bacterial content of Porphyromonas gingivalis, Prevotella intermedia, and Aggregatibacter actinomycetemcomitans using the modified Invader PLUS assay. The corresponding serum IgG titers were measured using ELISA. Changes in clinical parameters were evaluated over the course of 24 months. The sensitivity, specificity, and prediction values were calculated and used to determine cutoff points for prediction of the progression of chronic periodontitis. Results: Of the 124 individuals who completed the 24-month monitoring phase, 62 exhibited progression of periodontitis, whereas 62 demonstrated stable disease. The P. gingivalis counts of subgingival plaque from the deepest pockets was significantly associated with the progression of periodontitis (p < 0.001, positive predictive value = 0.708). Conclusions: The P. gingivalis counts of subgingival plaque from the deepest pockets may be associated with the progression of periodontitis..
2. Kohji Nozoe, Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Akira Haraguchi, Yoshitomi Aida, Fusanori Nishimura, Innate immune-stimulatory activity of Porphyromonas gingivalis fimbriae is eliminated by phase separation using Triton X-114, Journal of Immunological Methods, 10.1016/j.jim.2016.11.012, 441, 31-38, 2017.02, Fimbriae are virulence factors of Porphyromonas gingivalis (P. gingivalis). In this study, the action of fimbriae on neutrophil respiratory burst and cytokine production by mononuclear cells (MNC) were investigated. Native or denatured form of purified P. gingivalis fimbriae contained endotoxin at an equivalence of 1– 3 μg lipopolysaccharides (LPS)/mg protein. The endotoxin could be reduced to the equivalent of 1 ng-LPS/mg protein by phase separation using Triton X-114. Unfractionated fimbriae caused serum-dependent priming of neutrophils for enhanced respiratory burst, but both native and denatured forms of Triton X-114-fractionated fimbriae were not active at 100 μg/mL. Unfractionated fimbriae induced serum-dependent production of IL-1β by MNC. Triton X-114-fractionated fimbriae (10 μg/mL)-induced production of IL-1β, IL-8 or TNF-α was much lower than that induced by unfractionated fimbriae or 10 ng/mL P. gingivalis-LPS preparation. Triton X-114-fractionated fimbriae immobilized on polystyrene tubes induced adhesion-stimulated superoxide release by LPS-primed neutrophils in a β2 integrin-dependent manner. P. gingivalis cells caused priming of neutrophils; however, Toll-like receptor (TLR) 4 antagonists did not affect this response. Thus, P. gingivalis fimbriae were ineffective in inducing innate immune response in leukocytes; however, they induced β2 integrin-mediated response by neutrophils. Immune-stimulatory components of P. gingivalis might be recognized by receptors other than TLR4..
3. Masaaki Takeshita, Akira Haraguchi, Mayumi Miura, Takafumi Hamachi, Takao Fukuda, Terukazu Sanui, Aiko Takano, Fusanori Nishimura, Antibiotic effects against periodontal bacteria in organ cultured tissue, Clinical and Experimental Dental Research, 10.1002/cre2.48, 3, 1, 5-12, 2017.02, Mechanical reduction of infectious bacteria by using physical instruments is considered the principal therapeutic strategy for periodontal disease; addition of antibiotics is adjunctive. However, local antibiotic treatment, combined with conventional mechanical debridement, has recently been shown to be more effective in periodontitis subjects with type 2 diabetes. This suggests that some bacteria may invade the inflamed inner gingival epithelium, and mechanical debridement alone will be unable to reduce these bacteria completely. Therefore, we tried to establish infected organ culture models that mimic the inner gingival epithelium and aimed to see the effects of antibiotics in these established models. Mouse dorsal skin epithelia were isolated, and periodontal bacteria were injected into the epithelia. Infected epithelia were incubated with test antibiotics, and colony-forming ability was evaluated. Results indicated that effective antibiotics differed according to injected bacteria and the bacterial combinations tested. Overall, in organ culture model, the combination of amoxicillin or cefdinir and metronidazole compensate for the effects of less effective bacterial combinations on each other. This in vitro study would suggest effective periodontal treatment regimens, especially for severe periodontitis..
4. Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Akira Haraguchi, Yoshitomi Aida, Fusanori Nishimura, Anti-CD14 Antibody-treated Neutrophils Respond to LPS
Possible Involvement of CD14 Upregulated by Anti-CD14 Antibody Binding, Immunological Investigations, 10.1080/08820139.2016.1238925, 46, 2, 190-200, 2017.02, CD14 and Toll-like receptor 4/MD2 (TLR4/MD2) mediate the action of LPS on neutrophils. The anti-CD14 antibody and the TLR4/MD2-antagonist, synthetic lipid IVa (LA-14-PP), are known to inhibit the response of neutrophils to LPS. We studied the role of CD14 in LPS-induced priming of neutrophils for enhanced release of the superoxide anion. The anti-CD14 antibody at much higher concentrations than required to saturate CD14 was required to inhibit priming by LPS. The inhibitory effect of the anti-CD14 antibody was overcome by LPS. After washing, anti-CD14-treated neutrophils showed upregulated CD14 upon incubation at 37°C and responded to LPS with a delayed time-course. Thus, CD14-blocked neutrophils gained responsiveness to LPS through newly upregulated CD14. These results suggested that the unbound/free anti-CD14 antibody was essential to inhibit LPS-induced priming by blocking CD14 that were newly expressed during incubation at 37°C. LA-14-PP inhibited the response of neutrophils to LPS in an anti-CD14 antibody sensitive manner. When neutrophils were treated with LA-14-PP followed by treatment with the anti-CD14 antibody, CD14 was upregulated upon warming, but priming was blocked, suggesting that TLR4/MD2 was not newly expressed by warming in association with CD14 molecules. Thus, in addition to blocking CD14, the anti-CD14 antibody was found to induce the expression of new CD14..
5. Tomomi Sano, S. Nagayasu, S. Suzuki, Misaki Iwashita, Akiko Yamashita, T. Shinjo, Terukazu Sanui, A. Kushiyama, Takashi Kanematsu, T. Asano, Fusanori Nishimura, Epicatechin downregulates adipose tissue CCL19 expression and thereby ameliorates diet-induced obesity and insulin resistance, Nutrition, Metabolism and Cardiovascular Diseases, 10.1016/j.numecd.2016.11.008, 27, 3, 249-259, 2017.03, Background and aims Epicatechin (EC) intake has been suggested to be beneficial for the prevention of cardiovascular disorders, and it is well known that adipose tissue inflammation is one of the major risk factors for coronary heart diseases. The purpose of the present study was to determine the in vitro and in vivo effects of EC on adipose tissue inflammation and obesity. Methods and results DNA microarray analysis was performed to evaluate the effects of EC on gene expression in adipocytes co-cultured with bacterial endotoxin-stimulated macrophages. To determine the in vivo effects of the catechin, C57BL/6 mice were fed either a high-fat diet (HFD) or HFD combined with EC, and metabolic changes were observed EC suppressed the expression of many inflammatory genes in the adipocytes co-cultured with endotoxin-stimulated macrophages. Specifically, EC markedly suppressed chemokine (C–C motif) ligand 19 (CCL19) expression. The target cell of EC appeared to macrophages. The in vivo study indicated that mice fed the EC-supplemented HFD were protected from diet-induced obesity and insulin resistance. Accordingly, the expression levels of genes associated with inflammation in adipose tissue and in the liver were downregulated in this group of mice. Conclusions EC exerts beneficial effects for the prevention of adipose tissue inflammation and insulin resistance. Since we previously reported that mice deficient in the CCL19 receptor were protected from diet-induced obesity and insulin resistance, it can be concluded that the beneficial effects of EC could be mediated, at least in part, by marked suppression of CCL19 expression..
6. Aiko Takano, Takao Fukuda, Takanori Shinjo, Misaki Iwashita, Etsuko Matsuzaki, Kensuke Yamamichi, Masaaki Takeshita, Terukazu Sanui, Fusanori Nishimura, Angiopoietin-like protein 2 is a positive regulator of osteoblast differentiation, Metabolism: Clinical and Experimental, 10.1016/j.metabol.2017.01.006, 69, 157-170, 2017.04, Introduction and Aims Several studies have reported that angiopoietin-like protein 2 (Angptl2) is expressed abundantly in adipocytes and is associated with adipose tissue inflammation. In the present study, we found that osteoblasts and mesenchymal stem cells also expressed Angptl2 at high levels. The aim of this study was to understand the role of Angptl2 in osteoblastic cell differentiation. Methods Angptl2 expression was examined during osteoblast and adipocyte differentiation. The role of Angptl2 on cell differentiation and associated signaling was analyzed by gene knockdown using Angptl2 small interfering ribonucleic acid (siRNA). Results Angptl2 was highly expressed in MC3T3-E1 cells, ST2 cells and primary osteoblasts, but not in RAW264 cells. Inhibition of Angptl2 expression using siRNA markedly inhibited alkaline phosphatase (ALP) expression and osteoblastic differentiation in MC3T3-E1, ST2 cells and primary osteoblasts. Angptl2 siRNA also inhibited adipocyte differentiation in ST2 cells. Treatment of MC3T3-E1 cells with Angptl2 siRNA led to the down-regulation of the activities of several cell signaling pathways, including extracellular signal-regulated kinase (ERK), Jun amino-terminal kinase (JNK), Akt, and nuclear factor kappa B (NF-κB) signals. It also down-regulated the expression of Osterix, but not that of runt-related transcription factor 2 (Runx2), suggesting that Angptl2 is a positive activator of Osterix and its down-stream signals. Treatment of MC3T3-E1 cells with anti-Angptl2 antibodies suppressed ALP gene expression. In addition, treatment of Angptl2 siRNA-treated cells with culture supernatants of normal MC3T3-E1 cells restored ALP gene expression, indicating that Angptl2 acts in an autocrine manner. Conclusions The results suggest that Angptl2 is an autocrine positive regulator of cell differentiation. Thus, it is suggested that Angptl2 regulates not only adipose tissue metabolism but also bone metabolism..
7. Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Urara Tanaka, Rehab Alshargabi, Yoshitomi Aida, Fusanori Nishimura, Adhesion attenuates respiratory burst induced by different modes of triggering in resting or LPS-primed neutrophils, Immunobiology, 10.1016/j.imbio.2017.05.001, 222, 8-9, 865-871, 2017.08, The effects of adherence on neutrophil superoxide anion (O2 ) generation triggered by surface, soluble ligand, or adherence were studied. Resting-neutrophils adhered to the uncoated tubes resulting in O2 generation, but not on plasma-, fibrinogen-, vitronectin-, fibronectin-, laminin-, collagen-, or poly HEMA-coated surfaces. Enhanced N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated O2 generation by LPS-primed-neutrophils was induced by the incubation on plasma, fibrinogen, vitronectin, fibronectin, or laminin in the absence of Mg2+. In the presence of Mg2+, this response was observed in cells on collagen or poly HEMA. LPS-primed-neutrophils adhered to uncoated, BSA- or IgG-coated tubes and did not respond to fMLP, indicating that the fMLP-response of LPS-primed-neutrophils was suppressed by adherence. Upon incubation on plasma, fibrinogen, vitronectin, fibronectin in the presence of Mg2+, LPS-primed-neutrophils showed O2 generation. Upon incubation on collagen or poly HEMA, the primed-neutrophils neither generated O2 nor adhered. We found that O2 response of LPS-primed-neutrophils was attenuated depending on the time of exposure to plasma-coated surface. This attenuation was evident on plasma or fibrinogen, but not on collagen in the presence of Mg2+, indicating that O2 generation by LPS-primed-neutrophils was attenuated dependent on adherence but not on Mg2+. Thus, adhesion attenuated the O2 generation triggered by both soluble (fMLP) and insoluble (surface) stimuli..
8. Anthony C. May, Hiroshi Maeda, Hidemi Kurihara, Manabu Miyamoto, Hiroshi Hongyo, Ichiro Tanimoto, Atsushi Nagai, Fusanori Nishimura, Yoji Murayama, Keijiro Kato, Susumu Kokeguchi, Carla Cugini, Draft genome sequence of porphyromonas gingivalis strain 381 Okayama (381OKJP) stock culture, Microbiology Resource Announcements, 10.1128/MRA.01641-18, 8, 9, 2019.02, We report the draft genome sequence of Porphyromonas gingivalis strain 381 Okayama (381OKJP). The strain, obtained from the Socransky collection, has been used for experimentation since 1987. This sequence allows for comparisons to other sequenced 381 strains to observe acquisition of mutations and genome rearrangements in a commonly used laboratory strain..
9. Yusuke Nakatsu, Yasuka Matsunaga, Takeshi Yamamotoya, Koji Ueda, Masa ki Inoue, Yu Mizuno, Mikako Nakanishi, Tomomi Sano, Yosuke Yamawaki, Akifumi Kushiyama, Hideyuki Sakoda, Midori Fujishiro, Akihide Ryo, Hiraku Ono, Tohru Minamino, Shin Ichiro Takahashi, Haruya Ohno, Masayasu Yoneda, Kei Takahashi, Hisamitsu Ishihara, Hideki Katagiri, Fusanori Nishimura, Takashi Kanematsu, Tetsuya Yamada, Tomoichiro Asano, Prolyl Isomerase Pin1 Suppresses Thermogenic Programs in Adipocytes by Promoting Degradation of Transcriptional Co-activator PRDM16, Cell Reports, 10.1016/j.celrep.2019.02.066, 26, 12, 3221-3230.e3, 2019.03, Non-shivering thermogenesis in adipocytes provides defense against low temperatures and obesity development, but the underlying regulatory mechanism remains to be fully clarified. Based on both markedly increased Pin1 expression in states of excess nutrition and resistance to obesity development in Pin1 null mice, we speculated that adipocyte Pin1 may play a role in thermogenic programs. Adipose-specific Pin1 knockout (adPin1 KO) mice showed enhanced transcription of thermogenic genes and tolerance to hypothermia when exposed to cold. In addition, adPin1 KO mice were resistant to high-fat diet-induced obesity and glucose intolerance. A series of experiments revealed that Pin1 binds to PRDM16 and thereby promotes its degradation through the ubiquitin-proteasome system. Consistent with these results, Pin1 deletion in differentiated adipocytes showed enhancement of thermogenic programs in response to the β3 agonist CL316243 through the upregulation of PRDM16 proteins. These observations indicate that Pin1 is a negative regulator of non-shivering thermogenesis..
10. Shigeki Suzuki, Takao Fukuda, Shintaro Nagayasu, Jun Nakanishi, Kazuma Yoshida, Shizu Hirata-Tsuchiya, Yuki Nakao, Tomomi Sano, Akiko Yamashita, Satoru Yamada, Kouji Ohta, Hideki Shiba, Fusanori Nishimura, Dental pulp cell-derived powerful inducer of TNF-α comprises PKR containing stress granule rich microvesicles, Scientific reports, 10.1038/s41598-019-40046-2, 9, 1, 2019.03, It is well known that dental pulp tissue can evoke some of the most severe acute inflammation observed in the human body. We found that dental pulp cells secrete a factor that induces tumor necrosis factor-α production from macrophages, and designated this factor, dental pulp cell-derived powerful inducer of TNF-α (DPIT). DPIT was induced in dental pulp cells and transported to recipient cells via microvesicles. Treatment of dental pulp cells with a PKR inhibitor markedly suppressed DPIT activity, and weak interferon signals were constitutively activated inside the cells. In recipient macrophages, stimulation with DPIT-containing supernatants from pulp cells resulted in activation of both nuclear factor-κB and MAP kinases like JNK and p38. Proteomics analyses revealed that many stress granule-related proteins were present in supernatants from dental pulp cells as well as microvesicle marker proteins like GAPDH, β-actin, HSPA8, HSPB1, HSPE1, and HSPD1. Furthermore, giant molecule AHNAK and PKR were detected in microvesicles derived from dental pulp cells, and gene silencing of AHNAK in dental pulp cells led to reduced DPIT activity. Thus, it appeared that the core protein of DPIT was PKR, and that PKR was maintained in an active state in stress granule aggregates with AHNAK and transported via microvesicles. The activity of DPIT for TNF-α induction was far superior to that of gram-negative bacterial endotoxin. Therefore, we, report for the first time, that active PKR is transported via microvesicles as stress granule aggregates and induces powerful inflammatory signals in macrophages..
11. Hiroshi Nitta, Sayaka Katagiri, Toshiyuki Nagasawa, Yuichi Izumi, Isao Ishikawa, Hajime Izumiyama, Isao Uchimura, Masao Kanazawa, Hiroshige Chiba, Akira Matsuo, Kazunori Utsunomiya, Haruyasu Tanabe, Izumi Takei, Soichiro Asanami, Hiroshi Kajio, Toaki Ono, Yoichi Hayashi, Kiichi Ueki, Masatomi Tsuji, Yoichi Kurachi, Toshikazu Yamanouchi, Yoshimi Ichinokawa, Toshiki Inokuchi, Akiko Fukui, Shigeru Miyazaki, Takashi Miyauchi, Reiko Kawahara, Hideki Ogiuchi, Narihito Yoshioka, Jun Negishi, Masatomo Mori, Kenji Mogi, Yasushi Saito, Hideki Tanzawa, Tetsuo Nishikawa, Norihiko Takada, Kishio Nanjo, Nobuo Morita, Naoto Nakamura, Narisato Kanamura, Hirofumi Makino, Fusanori Nishimura, Kunihisa Kobayashi, Yoshinori Higuchi, Toshiie Sakata, Shigetaka Yanagisawa, Chuwa Tei, Yuichi Ando, Nobuhiro Hanada, Shuji Inoue, The number of microvascular complications is associated with an increased risk for severity of periodontitis in type 2 diabetes patients
Results of a multicenter hospital-based cross-sectional study, Journal of Diabetes Investigation, 10.1111/jdi.12633, 8, 5, 677-686, 2017.09, Aims/Introduction: To explore the relationships between periodontitis and microvascular complications as well as glycemic control in type 2 diabetes patients. Materials and Methods: This multicenter, hospital-based, cross-sectional study included 620 patients with type 2 diabetes. We compared the prevalence and severity of periodontitis between patients with ≥1 microvascular complication and those without microvascular complications. We also compared the prevalence and severity of periodontitis among patients with different degrees of glycemic control. Results: After adjusting for confounding factors, multiple logistic regression analysis showed that the severity of periodontitis was significantly associated with the number of microvascular complications (odds ratio 1.3, 95% confidence interval 1.1–1.6), glycated hemoglobin ≥8.0% (64 mmol/mol; odds ratio 1.6; 95% confidence interval 1.1–2.3), and older age (≥50 years; odds ratio 1.7; 95% confidence interval 1.1–2.6). However, the prevalence of periodontitis was not significantly associated with the number of microvascular complications, but was associated with male sex, high glycated hemoglobin (≥8.0% [64 mmol/mol]), older age (≥40 years), longer duration of diabetes (≥15 years) and fewer teeth (≤25). Furthermore, propensity score matching for age, sex, diabetes duration and glycated hemoglobin showed that the incidence of severe periodontitis was significantly higher among patients with microvascular complications than among those without microvascular complications (P < 0.05). Conclusions: The number of microvascular complications is a risk factor for more severe periodontitis in patients with type 2 diabetes, whereas poor glycemic control is a risk factor for increased prevalence and severity of periodontitis..
12. Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Urara Tanaka, Rehab Alshargabi, Yoshitomi Aida, Fusanori Nishimura, Roles of serum in innate immune responses of human leukocytes to synthetic lipopeptide, International Immunopharmacology, 10.1016/j.intimp.2017.06.006, 50, 61-68, 2017.09, Tripalmitoyl-S-glyceryl-L-Cys-Ser-(Lys)4 (Pam3CSK4) is a highly conserved molecular motif found in various classes of lipoproteins. The requirement for leukocyte to respond to synthetic Pam3CSK4 were studied. Pam3CSK4 primed neutrophils for a respiratory burst in a serum-dependent manner. Pam3CSK4 upregulated CD11b, CD14, and cytochrome b558, and downregulated Leu-8. Treatment of neutrophils with anti-CD14 antibodies and treatment of serum with anti-LPS binding protein (LBP) antibodies resulted in the inhibition of priming for respiratory burst by Pam3CSK4. It should be noted that LBP could not replicate the effects of serum in priming of neutrophils for respiratory burst by Pam3CSK4. Serum LBP bound to immobilized Pam3CSK4. Pam3CSK4 induced the interleukin-8 (IL-8) production by leukocytes in a serum-dependent manner. Further, Pam3CSK4-induced priming of neutrophils for respiratory burst was not inhibited by the LPS antagonists LA-14-PP, Rhodobacter sphaeroides LPS, or E5531, and Pam3CSK4-induced IL-8 production by leukocytes was not affected by LPS antagonist, E5531, indicating that Pam3CSK4 was recognized by a different receptor than LPS. Thus, Pam3CSK4 and LPS had similar biological activities and similar requirement to act on leukocytes, but were recognized by different receptors. Serum in the action of Pam3CSK4 on leukocytes was not replicated by LBP, suggesting that Pam3CSK4 might be disaggregated by serum to result in the activation of leukocytes..
13. Kensuke Yamamichi, Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Yuki Nakao, Karen Yotsumoto, Hiroaki Yamato, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Amelogenin induces M2 macrophage polarisation via PGE2/cAMP signalling pathway, Archives of Oral Biology, 10.1016/j.archoralbio.2017.08.005, 83, 241-251, 2017.11, Objectives Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. Design Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180). After performing a detailed microarray analysis, the effects of rM180 on macrophage phenotype and signal transduction pathways were evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, confocal microscopy and flow cytometry. Results The microarray analysis demonstrated that rM180 increased the expression of anti-inflammatory genes in lipopolysaccharide (LPS)-challenged macrophages after 24 h, while it temporarily up-regulated inflammatory responses at 4 h. rM180 significantly enhanced the expression of M2 macrophage markers (CD163 and CD206). rM180-induced M2 macrophage polarisation was associated with morphological changes as well as vascular endothelial growth factor (VEGF) production. rM180 enhanced prostaglandin E2 (PGE2) expression, and the activation of the cAMP/cAMP-responsive element binding (CREB) signaling pathway was involved in amelogenin-induced M2 macrophage polarisation. Blocking of PGE2 signaling by indomethacin specifically abrogated rM180 with or without LPS-induced M2 shift in PBMC-derived macrophages. Conclusion Amelogenin could reprogram macrophages into the anti-inflammatory M2 phenotype. It could therefore contribute to the early resolution of inflammation in periodontal lesions and provide a suitable environment for remodeling-periodontal tissues..
14. Yoshiaki Nomura, Toshiya Morozumi, Taneaki Nakagawa, Tsutomu Sugaya, Masamitsu Kawanami, Fumihiko Suzuki, Keiso Takahashi, Yuzo Abe, Soh Sato, Asako Makino-Oi, Atsushi Saito, Satomi Takano, Masato Minabe, Yohei Nakayama, Yorimasa Ogata, Hiroaki Kobayashi, Yuichi Izumi, Naoyuki Sugano, Koichi Ito, Satoshi Sekino, Yukihiro Numabe, Chie Fukaya, Nobuo Yoshinari, Mitsuo Fukuda, Toshihide Noguchi, Tomoo Kono, Makoto Umeda, Osamu Fujise, Fusanori Nishimura, Atsutoshi Yoshimura, Yoshitaka Hara, Toshiaki Nakamura, Kazuyuki Noguchi, Erika Kakuta, Nobuhiro Hanada, Shogo Takashiba, Yasuharu Amitani, Hiromasa Yoshie, Site-level progression of periodontal disease during a follow-up period, PLoS One, 10.1371/journal.pone.0188670, 12, 12, 2017.12, Periodontal disease is assessed and its progression is determined via observations on a site-by-site basis. Periodontal data are complex and structured in multiple levels; thus, applying a summary statistical approach (i.e., the mean) for site-level evaluations results in loss of information. Previous studies have shown the availability of mixed effects modeling. However, clinically beneficial information on the progression of periodontal disease during the follow-up period is not available. We conducted a multicenter prospective cohort study. Using mixed effects modeling, we analyzed 18,834 sites distributed on 3,139 teeth in 124 patients, and data were collected 5 times over a 24-month follow-up period. The change in the clinical attachment level (CAL) was used as the outcome variable. The CAL at baseline was an important determinant of the CAL changes, which varied widely according to the tooth surface. The salivary levels of periodontal pathogens, such as Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, were affected by CAL progression. “Linear”- and “burst”-type patterns of CAL progression occurred simultaneously within the same patient. More than half of the teeth that presented burst-type progression sites also presented linear-type progression sites, and most of the progressions were of the linear type. Maxillary premolars and anterior teeth tended to show burst-type progression. The parameters identified in this study may guide practitioners in determining the type and extent of treatment needed at the site and patient levels. In addition, these results show that prior hypotheses concerning "burst" and "linear" theories are not valid..
15. Takanori Shinjo, Fusanori Nishimura, Diabetes and periodontitis, Diabetes and Aging-related Complications, 10.1007/978-981-10-4376-5_15, 185-199, 2017.12, Periodontal disease is known as the sixth complication of diabetes. Recently, many clinical and epidemiologic reports have shown that local periodontal inflammation induces systemic micro-inflammation, contributing to insulin resistance and increasing the risk of cardiovascular diseases. We have used in vitro and in vivo studies to address the amplification mechanism of periodontal inflammation from the viewpoint of adipocyte-macrophage interaction. Our studies suggest that inflammatory cytokines, such as tumor necrosis factor-alpha (TNFα), are secreted from monocytes and macrophages that are stimulated by periodontal pathogen-derived Toll-like receptor ligands, such as lipopolysaccharides (LPS). TNFα then activates both adipocytes and infiltrated macrophages, thereby amplifying micro-inflammation through the synergistic production of inflammatory cytokines, including interleukin-6 and monocyte chemoattractant protein-1. Additionally, the expression of chemokine (C-C motif) ligand 19 (CCL19), involved in homing of dendritic cells, was found to be markedly upregulated in adipocytes co-cultured with LPS-stimulated macrophages. In vivo studies suggest that CCR7-CCL19 signaling possibly plays a critical role in adipose tissue metabolism through infiltration of immune cells, such as dendritic cells. Furthermore, the Hiroshima study, a clinical intervention study on diabetic patients receiving periodontal treatment, clearly showed that periodontal treatment combined with local oral antibiotic administration could improve glycated hemoglobin levels in subjects with a high-sensitivity C-reactive protein level > 500 ng/ml and a body mass index of approximately 25 kg/m2. A series of our studies suggests that periodontal treatment could improve glycemic control in diabetic patients with mild obesity..
16. Hiroshi Nitta, Sayaka Katagiri, Toshiyuki Nagasawa, Yuichi Izumi, Isao Ishikawa, Hajime Izumiyama, Isao Uchimura, Masao Kanazawa, Hiroshige Chiba, Akira Matsuo, Kazunori Utsunomiya, Haruyasu Tanabe, Izumi Takei, Soichiro Asanami, Hiroshi Kajio, Toaki Ono, Yoichi Hayashi, Kiichi Ueki, Masatomi Tsuji, Yoichi Kurachi, Toshikazu Yamanouchi, Yoshimi Ichinokawa, Toshiki Inokuchi, Akiko Fukui, Shigeru Miyazaki, Takashi Miyauchi, Reiko Kawahara, Hideki Ogiuchi, Narihito Yoshioka, Jun Negishi, Masatomo Mori, Kenji Mogi, Yasushi Saito, Hideki Tanzawa, Tetsuo Nishikawa, Norihiko Takada, Kishio Nanjo, Nobuo Morita, Naoto Nakamura, Narisato Kanamura, Hirofumi Makino, Fusanori Nishimura, Kunihisa Kobayashi, Yoshinori Higuchi, Toshiie Sakata, Shigetaka Yanagisawa, Chuwa Tei, Yuichi Ando, Nobuhiro Hanada, Shuji Inoue, The number of microvascular complications is associated with an increased risk for severity of periodontitis in type 2 diabetes patients
Results of a multicenter hospital-based cross-sectional study, Journal of Diabetes Investigation, 10.1111/jdi.12633, 8, 5, 677-686, 2017.09, Aims/Introduction: To explore the relationships between periodontitis and microvascular complications as well as glycemic control in type 2 diabetes patients. Materials and Methods: This multicenter, hospital-based, cross-sectional study included 620 patients with type 2 diabetes. We compared the prevalence and severity of periodontitis between patients with ≥1 microvascular complication and those without microvascular complications. We also compared the prevalence and severity of periodontitis among patients with different degrees of glycemic control. Results: After adjusting for confounding factors, multiple logistic regression analysis showed that the severity of periodontitis was significantly associated with the number of microvascular complications (odds ratio 1.3, 95% confidence interval 1.1–1.6), glycated hemoglobin ≥8.0% (64 mmol/mol; odds ratio 1.6; 95% confidence interval 1.1–2.3), and older age (≥50 years; odds ratio 1.7; 95% confidence interval 1.1–2.6). However, the prevalence of periodontitis was not significantly associated with the number of microvascular complications, but was associated with male sex, high glycated hemoglobin (≥8.0% [64 mmol/mol]), older age (≥40 years), longer duration of diabetes (≥15 years) and fewer teeth (≤25). Furthermore, propensity score matching for age, sex, diabetes duration and glycated hemoglobin showed that the incidence of severe periodontitis was significantly higher among patients with microvascular complications than among those without microvascular complications (P < 0.05). Conclusions: The number of microvascular complications is a risk factor for more severe periodontitis in patients with type 2 diabetes, whereas poor glycemic control is a risk factor for increased prevalence and severity of periodontitis..
17. Shigeki Suzuki, Hiroaki Hoshino, Kazuma Yoshida, Jun Nakanishi, Shizu Tsuchiya-Hirata, Seiji Kobuke, Naoto Haruyama, Fusanori Nishimura, Hideki Shiba, Genome-wide identification of chromatin-enriched RNA reveals that unspliced dentin matrix protein-1 mRNA regulates cell proliferation in squamous cell carcinoma, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.12.136, 495, 3, 2303-2309, 2018.01, Chromatin-enriched noncoding RNAs (ncRNAs) have emerged as key molecules in epigenetic processes by interacting with chromatin-associated proteins. Recently, protein-coding mRNA genes have been reported to be chromatin-tethered, similar with ncRNA. However, very little is known about whether chromatin-enriched mRNA is involved in the chromatin modification process. Here, we comprehensively examined chromatin-enriched RNA in squamous cell carcinoma (SQCC) cells by RNA subcellular localization analysis, which was a combination of RNA fractionation and RNA-seq. We identified 11 mRNAs as highly chromatin-enriched RNAs. Among these, we focused on the dentin matrix protein-1 (DMP-1) gene because its expression in SQCC cells has not been reported. Furthermore, we clarified that DMP-1 mRNA was retained in chromatin in its unspliced form in SQCC in vitro and in vivo. As the inhibition of the unspliced DMP-1 mRNA (unspDMP-1) expression resulted in decreased cellular proliferation in SQCC cells, we performed ChIP-qPCR to identify cell cycle-related genes whose expression was epigenetically modified by unspDMP-1, and found that the CDKN1B promoter became active in SQCC cells by inhibiting unspDMP-1 expression. This result was further validated by the increased CDKN1B gene expression in the cells treated with siRNA for unspDMP-1 and by restoration of the decreased cellular proliferation rate by simultaneously inhibiting CDKN1B expression in SQCC cells. Further, to examine whether unspDMP-1 was able to associate with the CDKN1B promoter region, SQCC cells stably expressing PP7-mCherry fusion protein were transiently transfected with the unspDMP-1 fused to 24 repeats of the PP7 RNA stem loop (unspDMP-1-24xPP7) and we found that unspDMP-1-24xPP7 was efficiently precipitated with the antibody against mCherry and was significantly enriched in the CDKN1B promoter region. Thus, unspDMP-1 is a novel chromatin-enriched RNA that epigenetically regulates cellular proliferation of SQCC..
18. Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Urara Tanaka, Rehab Alshargabi, Yoshitomi Aida, Fusanori Nishimura, Roles of serum in innate immune responses of human leukocytes to synthetic lipopeptide, International Immunopharmacology, 10.1016/j.intimp.2017.06.006, 50, 61-68, 2017.09, Tripalmitoyl-S-glyceryl-L-Cys-Ser-(Lys)4 (Pam3CSK4) is a highly conserved molecular motif found in various classes of lipoproteins. The requirement for leukocyte to respond to synthetic Pam3CSK4 were studied. Pam3CSK4 primed neutrophils for a respiratory burst in a serum-dependent manner. Pam3CSK4 upregulated CD11b, CD14, and cytochrome b558, and downregulated Leu-8. Treatment of neutrophils with anti-CD14 antibodies and treatment of serum with anti-LPS binding protein (LBP) antibodies resulted in the inhibition of priming for respiratory burst by Pam3CSK4. It should be noted that LBP could not replicate the effects of serum in priming of neutrophils for respiratory burst by Pam3CSK4. Serum LBP bound to immobilized Pam3CSK4. Pam3CSK4 induced the interleukin-8 (IL-8) production by leukocytes in a serum-dependent manner. Further, Pam3CSK4-induced priming of neutrophils for respiratory burst was not inhibited by the LPS antagonists LA-14-PP, Rhodobacter sphaeroides LPS, or E5531, and Pam3CSK4-induced IL-8 production by leukocytes was not affected by LPS antagonist, E5531, indicating that Pam3CSK4 was recognized by a different receptor than LPS. Thus, Pam3CSK4 and LPS had similar biological activities and similar requirement to act on leukocytes, but were recognized by different receptors. Serum in the action of Pam3CSK4 on leukocytes was not replicated by LBP, suggesting that Pam3CSK4 might be disaggregated by serum to result in the activation of leukocytes..
19. Hiroaki Matsunaga, Misaki Iwashita, Takanori Shinjo, Akiko Yamashita, Mitsudai Tsuruta, Shoichiro Nagasaka, Ataru Taniguchi, Mitsuo Fukushima, Naoya Watanabe, Fusanori Nishimura, Adipose tissue complement factor B promotes adipocyte maturation, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.11.069, 495, 1, 740-748, 2018.01, Objectives It is well-known that the complement system plays an essential role in host immunity. Observational studies have indicated that complement system-related molecules such as complement factor B (CfB) and other components are correlated with obesity and/or insulin resistance parameters. In this study, we investigated the role of adipocyte-derived CfB in adipose tissue metabolism. Methods We investigated the expression level of complement system-related genes in adipocytes. To understand the role of CfB in adipocyte, we performed Cfb overexpression in 3T3-L1 preadipocytes and generated adipocyte-specific Cfb transgenic mice. Results Cfb expression was markedly enhanced in 3T3-L1 adipocytes co-cultured with macrophages following endotoxin stimulation. In Cfb-overexpressing cells, the expression of adipocyte differentiation/maturation-related genes encoding peroxisome proliferator-activated receptor γ (Pparγ), adipocyte Protein 2 and perilipin was significantly enhanced. Cfb transgenic mice showed a marked increase in the expression of genes encoding Pparγ, perilipin, sterol regulatory element-binding protein 1 c, and Cd36 in the subcutaneous adipose tissue. Conclusions CfB plays a crucial role in late-phase of adipocyte differentiation and subsequent lipid droplet formation..
20. Kensuke Yamamichi, Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Yuki Nakao, Karen Yotsumoto, Hiroaki Yamato, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Amelogenin induces M2 macrophage polarisation via PGE2/cAMP signalling pathway, Archives of Oral Biology, 10.1016/j.archoralbio.2017.08.005, 83, 241-251, 2017.11, Objectives Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. Design Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180). After performing a detailed microarray analysis, the effects of rM180 on macrophage phenotype and signal transduction pathways were evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, confocal microscopy and flow cytometry. Results The microarray analysis demonstrated that rM180 increased the expression of anti-inflammatory genes in lipopolysaccharide (LPS)-challenged macrophages after 24 h, while it temporarily up-regulated inflammatory responses at 4 h. rM180 significantly enhanced the expression of M2 macrophage markers (CD163 and CD206). rM180-induced M2 macrophage polarisation was associated with morphological changes as well as vascular endothelial growth factor (VEGF) production. rM180 enhanced prostaglandin E2 (PGE2) expression, and the activation of the cAMP/cAMP-responsive element binding (CREB) signaling pathway was involved in amelogenin-induced M2 macrophage polarisation. Blocking of PGE2 signaling by indomethacin specifically abrogated rM180 with or without LPS-induced M2 shift in PBMC-derived macrophages. Conclusion Amelogenin could reprogram macrophages into the anti-inflammatory M2 phenotype. It could therefore contribute to the early resolution of inflammation in periodontal lesions and provide a suitable environment for remodeling-periodontal tissues..
21. Mitsudai Tsuruta, Misaki Iwashita, Takanori Shinjo, Hiroaki Matsunaga, Akiko Yamashita, Fusanori Nishimura, Metabolic Endotoxemia-Activated Macrophages Promote Pancreatic β Cell Death via IFNβ-Xaf1 Pathway, Hormone and Metabolic Research, 10.1055/s-0043-121467, 50, 2, 160-167, 2018.02, Metabolic endotoxemia has been implicated in the pathogenesis of type 2 diabetes. In addition to adipose tissue inflammation, inflammatory cell infiltration is also observed in islets, although its effect on islets is largely unknown. We hypothesized that macrophage infiltration into islets leads to impairment of α or β cell function, which ultimately act to exacerbate the pathophysiology of diabetes. Gene expression in a murine α cell line, αTC1, and β cell line, βTC6, was investigated by DNA microarray after co-culturing the cells with a murine macrophage cell line, RAW 264.7, in the presence or absence of bacterial endotoxin. Among the genes showing highly upregulated expression, genes specifically upregulated only in β cells were evaluated to determine the roles of the gene products on the cellular function of β cells. In both α and β cells, expression of type I interferon-responsive genes was highly upregulated upon endotoxin stimulation. Among these genes, expression of the X-linked inhibitor of apoptosis (Xiap)-associated factor 1 (Xaf1) gene, which is associated with the induction of apoptosis, was specifically enhanced in β cells by endotoxin stimulation. This upregulation appeared to be mediated by macrophage-derived interferon β (IFNβ), as endotoxin-stimulated macrophages produced higher amounts of IFNβ, and exogenous addition of IFNβ into βTC6 cultures resulted in increased Xaf1 protein production and cleaved caspase 3, which accelerated β-cell apoptosis. Macrophages activated by metabolic endotoxemia infiltrated into islets and produced IFNβ, which induced β-cell apoptosis by increasing the expression of Xaf1..
22. Yoshiaki Nomura, Toshiya Morozumi, Taneaki Nakagawa, Tsutomu Sugaya, Masamitsu Kawanami, Fumihiko Suzuki, Keiso Takahashi, Yuzo Abe, Soh Sato, Asako Makino-Oi, Atsushi Saito, Satomi Takano, Masato Minabe, Yohei Nakayama, Yorimasa Ogata, Hiroaki Kobayashi, Yuichi Izumi, Naoyuki Sugano, Koichi Ito, Satoshi Sekino, Yukihiro Numabe, Chie Fukaya, Nobuo Yoshinari, Mitsuo Fukuda, Toshihide Noguchi, Tomoo Kono, Makoto Umeda, Osamu Fujise, Fusanori Nishimura, Atsutoshi Yoshimura, Yoshitaka Hara, Toshiaki Nakamura, Kazuyuki Noguchi, Erika Kakuta, Nobuhiro Hanada, Shogo Takashiba, Yasuharu Amitani, Hiromasa Yoshie, Site-level progression of periodontal disease during a follow-up period, PLoS One, 10.1371/journal.pone.0188670, 12, 12, 2017.12, Periodontal disease is assessed and its progression is determined via observations on a site-by-site basis. Periodontal data are complex and structured in multiple levels; thus, applying a summary statistical approach (i.e., the mean) for site-level evaluations results in loss of information. Previous studies have shown the availability of mixed effects modeling. However, clinically beneficial information on the progression of periodontal disease during the follow-up period is not available. We conducted a multicenter prospective cohort study. Using mixed effects modeling, we analyzed 18,834 sites distributed on 3,139 teeth in 124 patients, and data were collected 5 times over a 24-month follow-up period. The change in the clinical attachment level (CAL) was used as the outcome variable. The CAL at baseline was an important determinant of the CAL changes, which varied widely according to the tooth surface. The salivary levels of periodontal pathogens, such as Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, were affected by CAL progression. “Linear”- and “burst”-type patterns of CAL progression occurred simultaneously within the same patient. More than half of the teeth that presented burst-type progression sites also presented linear-type progression sites, and most of the progressions were of the linear type. Maxillary premolars and anterior teeth tended to show burst-type progression. The parameters identified in this study may guide practitioners in determining the type and extent of treatment needed at the site and patient levels. In addition, these results show that prior hypotheses concerning "burst" and "linear" theories are not valid..
23. Sakiko Fujii, Katsumi Fujimoto, Noriko Goto, Yoshimitsu Abiko, Asayo Imaoka, Jinchang Shao, Kazuko Kitayama, Masami Kanawa, Agung Sosiawan, Ketut Suardita, Fusanori Nishimura, Yukio Kato, Characterization of human dental pulp cells grown in chemically defined Serum-Free medium, Biomedical Reports, 10.3892/br.2018.1066, 8, 4, 350-358, 2018.04, Dental pulp cells (DPCs) are promising candidates for use as transplantable cells in regenerative medicine. However, ex vivo expansion of these cells typically requires culture media containing fetal bovine serum, which may cause infection and immunological reaction following transplantation. In addition, the proliferation and differentiation of DPCs markedly depend upon serum batches. Therefore, the present study examined whether DPCs could be expanded under serum-free conditions. DPCs obtained from four donors were identified to proliferate actively in the serum-free medium, STK2, when compared with those cells in control medium (Dulbecco's modified Eagle's medium containing 10% serum). The high proliferative potential with STK2 was maintained through multiple successive culture passages. DNA microarray analyses demonstrated that the gene expression profile of DPCs grown in STK2 was similar to that of cells grown in the control medium; however, a number of genes related to cell proliferation, including placental growth factor and inhibin-βE, were upregulated in the STK2 cultures. Following induction of osteogenesis, DPCs grown in STK2 induced alkaline phosphatase activity and calcification at higher levels compared with the control medium cultures, indicating maintenance of differentiation potential in STK2. This serum-free culture system with DPCs may have applications in further experimental studies and as a clinical strategy in regenerative medicine..
24. Takanori Shinjo, Fusanori Nishimura, Diabetes and periodontitis, Diabetes and Aging-related Complications, 10.1007/978-981-10-4376-5_15, 185-199, 2017.12, Periodontal disease is known as the sixth complication of diabetes. Recently, many clinical and epidemiologic reports have shown that local periodontal inflammation induces systemic micro-inflammation, contributing to insulin resistance and increasing the risk of cardiovascular diseases. We have used in vitro and in vivo studies to address the amplification mechanism of periodontal inflammation from the viewpoint of adipocyte-macrophage interaction. Our studies suggest that inflammatory cytokines, such as tumor necrosis factor-alpha (TNFα), are secreted from monocytes and macrophages that are stimulated by periodontal pathogen-derived Toll-like receptor ligands, such as lipopolysaccharides (LPS). TNFα then activates both adipocytes and infiltrated macrophages, thereby amplifying micro-inflammation through the synergistic production of inflammatory cytokines, including interleukin-6 and monocyte chemoattractant protein-1. Additionally, the expression of chemokine (C-C motif) ligand 19 (CCL19), involved in homing of dendritic cells, was found to be markedly upregulated in adipocytes co-cultured with LPS-stimulated macrophages. In vivo studies suggest that CCR7-CCL19 signaling possibly plays a critical role in adipose tissue metabolism through infiltration of immune cells, such as dendritic cells. Furthermore, the Hiroshima study, a clinical intervention study on diabetic patients receiving periodontal treatment, clearly showed that periodontal treatment combined with local oral antibiotic administration could improve glycated hemoglobin levels in subjects with a high-sensitivity C-reactive protein level > 500 ng/ml and a body mass index of approximately 25 kg/m2. A series of our studies suggests that periodontal treatment could improve glycemic control in diabetic patients with mild obesity..
25. Soichiro Sonoda, Yu Feng Mei, Atsuta Ikiru, Atsushi Danjo, Haruyoshi Yamaza, Shion Hama, Kento Nishida, Ronghao Tang, Yukari Kyumoto, Norihisa Uehara, Toshio Kukita, Fusanori Nishimura, Takayoshi Yamaza, Exogenous nitric oxide stimulates the odontogenic differentiation of rat dental pulp stem cells, Scientific Reports, 10.1038/s41598-018-21183-6, 8, 1, 2018.12, Nitric oxide (NO) is thought to play a pivotal regulatory role in dental pulp tissues under both physiological and pathological conditions. However, little is known about the NO functions in dental pulp stem cells (DPSCs). We examined the direct actions of a spontaneous NO gas-releasing donor, NOC-18, on the odontogenic capacity of rat DPSCs (rDPSCs). In the presence of NOC-18, rDPSCs were transformed into odontoblast-like cells with long cytoplasmic processes and a polarized nucleus. NOC-18 treatment increased alkaline phosphatase activity and enhanced dentin-like mineralized tissue formation and the expression levels of several odontoblast-specific genes, such as runt related factor 2, dentin matrix protein 1 and dentin sialophosphoprotein, in rDPSCs. In contrast, carboxy-PTIO, a NO scavenger, completely suppressed the odontogenic capacity of rDPSCs. This NO-promoted odontogenic differentiation was activated by tumor necrosis factor-NF-κB axis in rDPSCs. Further in vivo study demonstrated that NOC-18-application in a tooth cavity accelerated tertiary dentin formation, which was associated with early nitrotyrosine expression in the dental pulp tissues beneath the cavity. Taken together, the present findings indicate that exogenous NO directly induces the odontogenic capacity of rDPSCs, suggesting that NO donors might offer a novel host DPSC-targeting alternative to current pulp capping agents in endodontics..
26. Shigeki Suzuki, Hiroaki Hoshino, Kazuma Yoshida, Jun Nakanishi, Shizu Tsuchiya-Hirata, Seiji Kobuke, Naoto Haruyama, Fusanori Nishimura, Hideki Shiba, Genome-wide identification of chromatin-enriched RNA reveals that unspliced dentin matrix protein-1 mRNA regulates cell proliferation in squamous cell carcinoma, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.12.136, 495, 3, 2303-2309, 2018.01, Chromatin-enriched noncoding RNAs (ncRNAs) have emerged as key molecules in epigenetic processes by interacting with chromatin-associated proteins. Recently, protein-coding mRNA genes have been reported to be chromatin-tethered, similar with ncRNA. However, very little is known about whether chromatin-enriched mRNA is involved in the chromatin modification process. Here, we comprehensively examined chromatin-enriched RNA in squamous cell carcinoma (SQCC) cells by RNA subcellular localization analysis, which was a combination of RNA fractionation and RNA-seq. We identified 11 mRNAs as highly chromatin-enriched RNAs. Among these, we focused on the dentin matrix protein-1 (DMP-1) gene because its expression in SQCC cells has not been reported. Furthermore, we clarified that DMP-1 mRNA was retained in chromatin in its unspliced form in SQCC in vitro and in vivo. As the inhibition of the unspliced DMP-1 mRNA (unspDMP-1) expression resulted in decreased cellular proliferation in SQCC cells, we performed ChIP-qPCR to identify cell cycle-related genes whose expression was epigenetically modified by unspDMP-1, and found that the CDKN1B promoter became active in SQCC cells by inhibiting unspDMP-1 expression. This result was further validated by the increased CDKN1B gene expression in the cells treated with siRNA for unspDMP-1 and by restoration of the decreased cellular proliferation rate by simultaneously inhibiting CDKN1B expression in SQCC cells. Further, to examine whether unspDMP-1 was able to associate with the CDKN1B promoter region, SQCC cells stably expressing PP7-mCherry fusion protein were transiently transfected with the unspDMP-1 fused to 24 repeats of the PP7 RNA stem loop (unspDMP-1-24xPP7) and we found that unspDMP-1-24xPP7 was efficiently precipitated with the antibody against mCherry and was significantly enriched in the CDKN1B promoter region. Thus, unspDMP-1 is a novel chromatin-enriched RNA that epigenetically regulates cellular proliferation of SQCC..
27. Hiroaki Matsunaga, Misaki Iwashita, Takanori Shinjo, Akiko Yamashita, Mitsudai Tsuruta, Shoichiro Nagasaka, Ataru Taniguchi, Mitsuo Fukushima, Naoya Watanabe, Fusanori Nishimura, Adipose tissue complement factor B promotes adipocyte maturation, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.11.069, 495, 1, 740-748, 2018.01, Objectives It is well-known that the complement system plays an essential role in host immunity. Observational studies have indicated that complement system-related molecules such as complement factor B (CfB) and other components are correlated with obesity and/or insulin resistance parameters. In this study, we investigated the role of adipocyte-derived CfB in adipose tissue metabolism. Methods We investigated the expression level of complement system-related genes in adipocytes. To understand the role of CfB in adipocyte, we performed Cfb overexpression in 3T3-L1 preadipocytes and generated adipocyte-specific Cfb transgenic mice. Results Cfb expression was markedly enhanced in 3T3-L1 adipocytes co-cultured with macrophages following endotoxin stimulation. In Cfb-overexpressing cells, the expression of adipocyte differentiation/maturation-related genes encoding peroxisome proliferator-activated receptor γ (Pparγ), adipocyte Protein 2 and perilipin was significantly enhanced. Cfb transgenic mice showed a marked increase in the expression of genes encoding Pparγ, perilipin, sterol regulatory element-binding protein 1 c, and Cd36 in the subcutaneous adipose tissue. Conclusions CfB plays a crucial role in late-phase of adipocyte differentiation and subsequent lipid droplet formation..
28. Mitsudai Tsuruta, Misaki Iwashita, Takanori Shinjo, Hiroaki Matsunaga, Akiko Yamashita, Fusanori Nishimura, Metabolic Endotoxemia-Activated Macrophages Promote Pancreatic β Cell Death via IFNβ-Xaf1 Pathway, Hormone and Metabolic Research, 10.1055/s-0043-121467, 50, 2, 160-167, 2018.02, Metabolic endotoxemia has been implicated in the pathogenesis of type 2 diabetes. In addition to adipose tissue inflammation, inflammatory cell infiltration is also observed in islets, although its effect on islets is largely unknown. We hypothesized that macrophage infiltration into islets leads to impairment of α or β cell function, which ultimately act to exacerbate the pathophysiology of diabetes. Gene expression in a murine α cell line, αTC1, and β cell line, βTC6, was investigated by DNA microarray after co-culturing the cells with a murine macrophage cell line, RAW 264.7, in the presence or absence of bacterial endotoxin. Among the genes showing highly upregulated expression, genes specifically upregulated only in β cells were evaluated to determine the roles of the gene products on the cellular function of β cells. In both α and β cells, expression of type I interferon-responsive genes was highly upregulated upon endotoxin stimulation. Among these genes, expression of the X-linked inhibitor of apoptosis (Xiap)-associated factor 1 (Xaf1) gene, which is associated with the induction of apoptosis, was specifically enhanced in β cells by endotoxin stimulation. This upregulation appeared to be mediated by macrophage-derived interferon β (IFNβ), as endotoxin-stimulated macrophages produced higher amounts of IFNβ, and exogenous addition of IFNβ into βTC6 cultures resulted in increased Xaf1 protein production and cleaved caspase 3, which accelerated β-cell apoptosis. Macrophages activated by metabolic endotoxemia infiltrated into islets and produced IFNβ, which induced β-cell apoptosis by increasing the expression of Xaf1..
29. Tomoyuki Kurashina, Shoichiro Nagasaka, Naoya Watanabe, Daisuke Yabe, Noriko Sugi, Kazuko Nin, Masaya Hosokawa, Yoshio Nomura, Mitsuo Fukushima, Yoshikatsu Nakai, Fusanori Nishimura, Ataru Taniguchi, Circulating TNF receptor 2 is closely associated with the kidney function in non-diabetic Japanese subjects, Journal of Atherosclerosis and Thrombosis, 10.5551/jat.21055, 21, 7, 730-738, 2014.01, Aim: Chronic kidney disease (CKD) is associated with cardiovascular events. Tumor necrosis factor (TNF) and/or its receptors have been postulated to be involved in renal pathophysiology. It is unclear whether an increased TNF system activity is present before the development of apparent CKD. Methods: Four hundred and twenty non-diabetic Japanese subjects with an estimated GFR (eGFR) greater than 60 ml/min/1.73 m2 were recruited for measurement of the HbA1c, insulin, TNF system activity (TNF-α, soluble TNF receptor 1 (sTNF-R1) and sTNF-R2) levels and various parameters, including the lipid, high-sensitivity C-reactive protein (hsCRP), high-molecular-weight (HMW) adiponectin and leptin levels. The subjects were stratified according to the eGFR: the G1 level (eGFR ≥90 ml/min/1.73 m2) and the G2 level (90 2). Results: Whereas no significant differences were observed in gender, body mass index (BMI), blood pressure, insulin, TNF-α, hsCRP, HMW adiponectin or leptin between the two groups, the values for age, HbA1c, triglycerides, sTNF-R1 and sTNF-R2 were significantly higher in the subjects with a G2 level of eGFR than in those with a G1 level. In contrast, the HDL cholesterol levels were significantly lower in the subjects with a G2 level than in those with a G1 level. Linear negative correlations were also observed between eGFR and age, BMI, HbA1c, triglycerides, sTNF-R1 and sTNFR2, respectively. A multiple logistic regression analysis revealed that only sTNF-R2 was associated with the presence of a G2 level of eGFR (Odds ratio 1.092, 95% CI 1.013-1.177, P= 0.021). Conclusions: The circulating sTNF-R2 level is closely associated with the kidney function in nondiabetic Japanese subjects..
30. K. Hisano, O. Fujise, M. Miura, T. Hamachi, E. Matsuzaki, Fusanori Nishimura, The pga gene cluster in Aggregatibacter actinomycetemcomitans is necessary for the development of natural competence in Ca2+-promoted biofilms, Molecular Oral Microbiology, 10.1111/omi.12046, 29, 2, 79-89, 2014.04, Natural competence is the ability of bacteria to incorporate extracellular DNA into their genomes. This competence is affected by a number of factors, including Ca2+ utilization and biofilm formation. As bacteria can form thick biofilms in the presence of extracellular Ca2+, the additive effects of Ca2+-promoted biofilm formation on natural competence should be examined. We evaluated natural competence in Aggregatibacter actinomycetemcomitans, an important periodontal pathogen, in the context of Ca2+-promoted biofilms, and examined whether the pga gene cluster, required for bacterial cell aggregation, is necessary for competence development. The A. actinomycetemcomitans cells grown in the presence of 1 mm CaCl2 exhibited enhanced cell aggregation and increased levels of cell-associated Ca2+. Biofilm-derived cells grown in the presence of Ca2+ exhibited the highest levels of natural transformation frequency and enhanced expression of the competence regulator gene, tfoX. Natural competence was enhanced by the additive effects of Ca2+-promoted biofilms, in which high levels of pga gene expression were also detected. Mutation of the pga gene cluster disrupted biofilm formation and competence development, suggesting that these genes play a critical role in the ability of A. actinomycetemcomitans to adapt to its natural environment. The Ca2+-promoted biofilms may enhance the ability of bacteria to acquire extracellular DNA..
31. Yuko Sugiura, Yoshihiko Soga, Ichiro Tanimoto, Susumu Kokeguchi, Sachiko Morishige-Nishide, Kotoe Itami-Kono, Kanayo Takahashi, Nobuharu Fujii, Fumihiko Ishimaru, Mitsune Tanimoto, Kokoro Yamabe, Soichiro Tsutani, Fusanori Nishimura, Shogo Takashiba, With regard to our manuscripts on the commercial saliva substitute, Oralbalance®—its formula has been changed, Supportive Care in Cancer, 10.1007/s00520-014-2432-8, 22, 12, 3121-3122, 2014.11.
32. Shigeki Suzuki, Seiji Kobuke, Naoto Haruyama, Hiroaki Hoshino, Ashok B. Kulkarni, Fusanori Nishimura, Adhesive and migratory effects of phosphophoryn are modulated by flanking peptides of the integrin binding motif, PLoS One, 10.1371/journal.pone.0112490, 9, 11, 2014.11, Phosphophoryn (PP) is generated from the proteolytic cleavage of dentin sialophosphoprotein (DSPP). Gene duplications in the ancestor dentin matrix protein-1 (DMP-1) genomic sequence created the DSPP gene in toothed animals. PP and DMP-1 are phosphorylated extracellular matrix proteins that belong to the family of small integrin-binding ligand N-linked glycoproteins (SIBLINGs). Many SIBLING members have been shown to evoke various cell responses through the integrinbinding Arg-Gly-Asp (RGD) domain; however, the RGD-dependent function of PP is not yet fully understood. We demonstrated that recombinant PP did not exhibit any obvious cell adhesion ability, whereas the simultaneously purified recombinant DMP-1 did. A cell adhesion inhibitory analysis was performed by pre-incubating human osteosarcoma MG63 cells with various PP peptides before seeding onto vitronectin. The results obtained revealed that the incorporation of more than one amino acid on both sides of the PP-RGD domain was unable to inhibit the adhesion of MG63 cells onto vitronectin. Furthermore, the inhibitory activity of a peptide containing the PP-RGD domain with an open carboxyl-terminal side (H-463SDESDTNSESANESGSRGDA482-OH) was more potent than that of a peptide containing the RGD domain with an open amino-terminal side (H-478SRGDASYTSDESSDDDNDSDSH499-OH). This phenomenon was supported by the potent cell adhesion and migration abilities of the recombinant truncated PP, which terminated with Ala482. Furthermore, various point mutations in Ala482 and/or Ser483 converted recombinant PP into cell-adhesive proteins. Therefore, we concluded that the Ala482-Ser483 flanking sequence, which was detected in primates and mice, was the key peptide bond that allowed the PPRGD domain to be sequestered. The differential abilities of PP and DMP-1 to act on integrin imply that DSPP was duplicated from DMP-1 to serve as a crucial extracellular protein for tooth development rather than as an integrin-mediated signaling molecule..
33. Hirofumi Okubo, Yusuke Nakatsu, Hideyuki Sakoda, Akifumi Kushiyama, Midori Fujishiro, Toshiaki Fukushima, Yasuka Matsunaga, Haruya Ohno, Masayasu Yoneda, Hideaki Kamata, Takanori Shinjo, Misaki Iwashita, Fusanori Nishimura, Tomoichiro Asano, Mosapride citrate improves nonalcoholic steatohepatitis with increased fecal lactic acid bacteria and plasma glucagon-like peptide-1 level in a rodent model, American Journal of Physiology - Gastrointestinal and Liver Physiology, 10.1152/ajpgi.00198.2014, 308, 2, G151-G158, 2015.01, Several lines of evidence have suggested a role of gut microbiota in the etiology of nonalcoholic steatohepatitis (NASH). NASH subjects reportedly showed a prolonged orocecal transit time coexistent with small intestinal bacterial overgrowth. We considered the possibility that enhanced gastrointestinal motility would influence gut microbiota and thus investigated the effects of the gastroprokinetic agent mosapride citrate (MC) on gut microbiota and the development of NASH using a methionine-choline deficient (MCD) diet-fed rodent model. Mice were divided into three groups, given the normal chow diet (NCD), the MCD diet, or the MCD diet containing 10 mg·kg–1·day–1 of MC (MCD plus MC) for 6 wk. NASH development was evaluated based on hepatic histochemical findings, serum parameters and various mRNA and/or protein expression levels. MC treatment suppressed MCD diet-induced NASH development, with reduced serum lipopoly-saccharide and increased plasma glucagon-like peptide-1 (GLP-1) concentrations. Calculation of the relative abundance of each strain based on gut microbiota analyses indicated lactic acid bacteria specifically, such as Bifidobacterium and Lactobacillus, in feces to be decreased in the MCD, compared with the NCD group. Interestingly, the reduction in lactic acid bacteria in the MCD diet group was reversed in the MCD plus MC group. In addition, colon inflammation observed in the MCD diet group was reduced in the MCD plus MC group. Therefore, MC showed a protective effect against MCD diet-induced NASH development in our rodent model, with possible involvements of increased fecal lactic acid bacteria, protection against colon inflammation and elevated plasma GLP-1..
34. Michiko Furuta, Yoshihiro Shimazaki, Shunichi Tanaka, Kenji Takeuchi, Yukie Shibata, Toru Takeshita, Fusanori Nishimura, Yoshihisa Yamashita, Gender-specific associations of serum antibody to Porphyromonas gingivalis and inflammatory markers, BioMed Research International, 10.1155/2015/897971, 2015, 2015.01, It remains unclear whether serum antibody titer against Porphyromonas gingivalis (Pg) and inflammatory components lead to periodontal deterioration in each gender, as periodontal and systemic status is influenced by gender. The present study investigates the gender-specific probable effects of titer against Pg and inflammatory markers on periodontal health status in a longitudinal study. A retrospective study design was used. At two time points over an 8-year period (in 2003 and 2011), 411 individuals (295 males with a mean age of 57.6 ± 11.2 years and 116 females with a mean age of 59.2 ± 10.3 years) were surveyed. Periodontal status, serum antibody titer against Pg, and high-sensitive C-reactive protein (hsCRP) were evaluated. Poisson regression analyses revealed that the elevated titer against Pg and hsCRP significantly predicted the persistence of periodontal disease 8 years later in females with periodontal disease in 2003. Elevated hsCRP was significantly associated with the incidence of periodontal disease 8 years later in females who were periodontally healthy in 2003. Males had a weaker association among titer against Pg, inflammatory markers, and periodontal disease. These findings suggest that immune response to Pg infection in addition to inflammatory components affects periodontal deterioration in females..
35. Takanori Shinjo, Yusuke Nakatsu, Misaki Iwashita, Tomomi Sano, Hideyuki Sakoda, Hisamitsu Ishihara, Akifumi Kushiyama, Midori Fujishiro, Toshiaki Fukushima, Yoshihiro Tsuchiya, Hideaki Kamata, Fusanori Nishimura, Tomoichiro Asano, DPP-IV inhibitor anagliptin exerts anti-inflammatory effects on macrophages, adipocytes, and mouse livers by suppressing NF-κB activation, American Journal of Physiology - Heart and Circulatory Physiology, 10.1152/ajpendo.00553.2014, 309, 3, E214-E223, 2015.01, Dipeptidyl peptidase IV (DPPIV) expression in visceral adipose tissue is reportedly increased in obese patients, suggesting an association of DPP-IV with inflammation. In this study, first, lipopolysaccharide (LPS)- or palmitateinduced elevations of inflammatory cytokine mRNA expressions in RAW264.7 macrophages were shown to be significantly suppressed by coincubation with a DPP-IV inhibitor, anagliptin (10 μM), despite low DPP-IV expression in the RAW264.7 cells. Regarding the molecular mechanism, LPS-induced degradation of IκBα and phosphorylations of p65, JNK, and p38, as well as NF- κB and AP-1 promoter activities, were revealed to be suppressed by incubation with anagliptin, indicating suppressive effects of anagliptin on both NF- κB and AP-1 signaling pathways. Anagliptin also acted on 3T3-L1 adipocytes, weakly suppressing the inflammatory cytokine expressions induced by LPS and TNFα. When 3T3-L1 and RAW cells were cocultured and stimulated with LPS, the effects of anagliptin on the suppression of cytokine expressions in 3T3-L1 adipocytes were more marked and became evident at the 10 μM concentration. Anti-inflammatory effects of anagliptin were also observed in vivo on the elevated hepatic and adipose expressions and serum concentrations of inflammatory cytokines in association with the suppression of hepatic NF- κB transcriptional activity in LPS-infused mice. Taking these observations together, the anti-inflammatory properties of anagliptin may be beneficial in terms of preventing exacerbation of diabetes and cardiovascular events..
36. Terukazu Sanui, Urara Tanaka, Takao Fukuda, Kyousuke Toyoda, Takaharu Taketomi, Ryo Atomura, Kensuke Yamamichi, Fusanori Nishimura, Mutation of Spry2 Induces Proliferation and Differentiation of Osteoblasts but Inhibits Proliferation of Gingival Epithelial Cells, Journal of Cellular Biochemistry, 10.1002/jcb.25014, 116, 4, 628-639, 2015.04, Sprouty was identified as an inhibitor of the fibroblast growth factor (FGF) receptor, and Sprouty2 (Spry2) functions as a negative regulator of receptor tyrosine kinase signaling. In this study, we investigated how inhibition of Spry2 affects osteoblasts and gingival epithelial cells in periodontal tissue regeneration in vitro. Transduction of a dominant-negative mutant of Spry2 (Y55A-Spry2) enhanced basic fibroblast growth factor (bFGF)- and epidermal growth factor (EGF)-induced ERK activation in MC3T3-E1 osteoblastic cells. In contrast, it decreased their activation in GE1 cells. Consistent with these observations, Y55A-Spry2 increased osteoblast proliferation with bFGF and EGF stimulation, whereas the proliferation of Y55A-Spry2-introduced GE1 cells was decreased via the ubiquitination and degradation of EGF receptors (EGFRs). In addition, Y55A-Spry2 caused upregulation of Runx2 expression and downregulation of Twist, a negative regulator of Runx2, with treatment of bFGF and EGF, resulting in enhanced osteoblastogenesis accompanied by alkaline phosphatase activation and osteocalcin expression in MC3T3-E1 cells. These data suggest that suppression of Spry2 expression induces proliferation and differentiation of osteoblastic cells after the addition of a bFGF and EGF cocktail but inhibits proliferation in gingival epithelial cells. These in vitro experiments may provide a molecular basis for novel therapeutic approaches in periodontal tissue regeneration. Taken together, our study proposes that combined application of an inhibitor for tyrosine 55 of Spry2, bFGF, and EGF may effectively allow alveolar bone growth and block the ingrowth of gingival epithelial cells toward bony defects, biologically mimicking a barrier effect in guided tissue regeneration, with in vivo investigation in the future. J. Cell. Biochem. 116: 628-639, 2015..
37. Seiji Kobuke, Shigeki Suzuki, Hiroaki Hoshino, Naoto Haruyama, Fusanori Nishimura, Hideki Shiba, Relationship between length variations in Ser/Asp-rich repeats in phosphophoryn and in vitro precipitation of calcium phosphate, Archives of Oral Biology, 10.1016/j.archoralbio.2015.05.013, 60, 9, 1263-1272, 2015.06, Objective Phosphophoryn (PP) is generated from the proteolytic cleavage of dentin sialophosphoprotein (DSPP). PP which contains tandem serine/asparatic acid rich repeats (SDrr) is known to enhance dentin mineralization. The nucleotide sequences coding SDrr are identified in the DSPP genes of toothed animals and the length variations of SDrr between intra- and inter-species have been reported. However, it remains unknown about the relationship between the length variations in SDrr and the functions of PP in matrix mineralization. Design By utilizing a mammalian expression system, we generated several recombinant PP proteins (rPP) containing SDrr of different lengths and analyzed their effects on the precipitation of calcium phosphate with an in vitro gel diffusion system. Results rPP-Δ37.6 SDrr and rPP-Δ63.5 SDrr, which possessed shortened SDrr that accounted for 62.4 and 36.5% the length of SDrr in full-length rPP (rPP-full), respectively, induced the precipitation of calcium phosphate similar to that of rPP-full at the same molar concentration, whereas rPP-ΔSDrr, in which SDrr were flipped, did not. Furthermore, rPP-Δ63.5 SDrr significantly increased the accumulation of calcium compared with rPP-full at adjusted concentrations so that the same amounts of SDrr were embedded. The results of an ELISA analysis indicated that the amounts of rPP-Δ37.6 SDrr and rPP-Δ63.5 SDrr secreted from transfected cells were 5.2- and 7.1-fold greater than that of rPP-full, respectively. Conclusions The generated rPP-Δ63.5 SDrr which can be substituted for rPP-full may be a candidate for a therapeutic molecule to facilitate hard tissue generation such as reparative dentin formation..
38. Takanori Shinjo, Yusuke Nakatsu, Misaki Iwashita, Tomomi Sano, Hideyuki Sakoda, Hisamitsu Ishihara, Akifumi Kushiyama, Midori Fujishiro, Fusanori Nishimura, Tomoichiro Asano, High-fat diet feeding significantly attenuates anagliptin-induced regeneration of islets of Langerhans in streptozotocin-induced diabetic mice, Diabetology and Metabolic Syndrome, 10.1186/s13098-015-0047-y, 7, 1, 2015.06, Background: DPP-4 inhibitors reportedly exert effects on both alpha and beta cells, and promote the proliferation and survival of beta cells. We investigated the effects of anagliptin on structurally-impaired islets of Langerhans in streptozotocin (STZ)-treated mice, fed either a normal or a high-fat diet. Pdx-1 expression in the pancreas and serum insulin/glucagon concentrations were also examined. Findings: Anagliptin treatment significantly up-regulated pancreatic Pdx-1 expression, with elevated serum glucagon-like peptide-1 concentrations, regardless of whether the diet was normal or high-fat. However, interestingly, the beta cell regeneration, structural normalization of islets of Langerhans including alpha cell: beta cell area ratios, and serum insulin elevation, all observed with anagliptin administration in the animals fed a normal diet, were markedly suppressed in the high-fat fed group. Conclusions: High-fat diet feeding clearly weakened the regenerative effects of anagliptin on the islets of Langerhans in STZ-treated mice. Our findings suggest the importance of normalizing lipid metabolism for full manifestation of DPP-4 inhibitor effects on the islets of Langerhans..
39. Fusanori NISHIMURA, Angiopoietin-like protein 2 is a positive regulator of osteoblast differentiation. , Metabolism, 2017.01.
40. Fusanori NISHIMURA, Innate immune-stimulatory activity of Porphyromonas gingivalis fimbriae is eliminated by phase separation using Triton X-114., J Immunol Methods, 2016.11.
41. Fusanori NISHIMURA, Anti-CD14 Antibody-treated Neutrophils Respond to LPS: Possible Involvement of CD14 Upregulated by Anti-CD14 Antibody Binding., Immunol Invest, 1-11, 2017.11.
42. Fusanori NISHIMURA, Epicatechin downregulates adipose tissue CCL19 expression and thereby ameliorates diet-induced obesity and insulin resistance, Nutr Metab Cardiovasc, 10.1016/j.numecd.2016.11.00, 27, 3, 249-259, 2016.11.
43. Fusanori NISHIMURA, Sphingosine-1-phosphate/S1PR2-mediated signaling triggers Smad1/5/8 phosphorylation and thereby induces Runx2 expression in osteoblasts., Bone, 10.1016/j.bone.2016.09.003, 93, 1-11, 2016.09.
44. Fusanori NISHIMURA, Sphingosine-1-phosphate-enhanced Wnt5a promotes osteogenic differentiation in C3H10T1/2 cells. , Cell Biol Int, 10.1002/cbin.10652., 40, 10, 1129-2236, 2016.10.
45. Fusanori NISHIMURA, IL-17A synergistically enhances TNFα-induced IL-6 and CCL20 production in 3T3-L1 adipocytes., Biochem Biophys Res Commun, 10.1016/j.bbrc.2016.06.049, 477, 2, 2016.04.
46. Fusanori NISHIMURA, Inhibition of Sprouty2 polarizes macrophages toward an M2 phenotype by stimulation with interferon γ and Porphyromonas gingivalis lipopolysaccharide., Immun Inflamm Dis, 10.1002/iid3.99., 2016.03.
47. Fusanori NISHIMURA, Mechanisms of the Macrolide-Induced Inhibition of Superoxide Generation by Neutrophils., Inflammation, 39, 3, 1039-1048, 2017.06.
48. Fusanori NISHIMURA, Disaggregation of lipopolysaccharide by albumin, hemoglobin, or high density lipoprotein, forming complexes that prime neutrophils for enhanced release of superoxide., Pathog Dis, 2016.03.
49. Fusanori NISHIMURA, Interferon-gamma improves impaired dentinogenic and immunosuppressive functions of irreversible pulpitis-derived human dental pulp stem cells., Sci Rep, 10.1038/srep19286, 2016., 6, 19286-19286, 2016.06.
50. Fusanori NISHIMURA, Sprouty2 inhibition promotes proliferation and migration of periodontal ligament cells., Oral Dis, 10.1111/odi.12369, 21, 977-986, 2015.10.
51. Fusanori NISHIMURA, Grp78 is Critical for Amelogenin-Induced Cell Migration in a Multipotent Clonal Human Periodontal Ligament Cell Line., J Cell Physiol, 10.1002/jcp.25087., [Epub ahead of print], 2015.12.
52. Fusanori NISHIMURA, High-fat diet feeding significantly attenuates anagliptin-induced regeneration of islets of Langerhans in streptozotocin-induced diabetic mice., Diabetol Metab Syndr, 10.1186/s13098-015-0047-y, 7, 50, 2015.07.
53. Fusanori NISHIMURA, DPP-4 inhibitor anagliptin exerts anti-inflammatory effects on macrophages, adipocytes, and mouse livers by suppressing NF-κB activation., Am J Physiol Endocrinol Metab, 10.1152/ajpendo.00553.2014, 309, 3, E214-E223, 2015.04.
54. Fusanori NISHIMURA, Protection from diet-induced obesity and insulin resistance in mice lacking CCL19-CCR7 signaling., Obesity, 10.1002/oby.21127, 23, 7, 1460-1471, 2015.07.
55. Fusanori NISHIMURA, Mutation of Spry2 induces proliferation and differentiation of osteoblasts but inhibits proliferation of gingival epithelial cells., J Cell Biochem, 10.1002/jcb.25014., 116, 4, 628-639, 2015.04.
56. Fusanori NISHIMURA, Sphingosine-1-phosphate inhibits differentiation of C3H10T1/2 cells into adipocyte., Mol Cell Biochem, 10.1007/s11010-014-2290-1., 2015.03.
57. Fusanori NISHIMURA, Mosapride citrate improves non-alcoholic steatohepatitis with increased fecal lactic acid bacteria and plasma glucagon-like peptide-1 level in a rodent model., Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00198.2014, 2015.01.
58. Fusanori NISHIMURA, Adhesive and migratory effects of phosphophoryn are modulated by flanking peptides of the integrin binding motif, , PLOS One, 10.1371/journal.pone.0112490, 2014.09.
59. Fusanori NISHIMURA, Lactobacillus casei strain Shirota protects against non-alcoholic steatohepatitis development in a rodent model., Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00225.2013., 305, 12, G911-G918, 2013.12.
60. Fusanori NISHIMURA, The Inflammation-lipocalin2 axis may contribute to the development of chronic kidney disease., Nephrol Dial Transplant, 10.1093/ndt/gft449. , 29, 3, 611-618, 2014.03.
61. Fusanori NISHIMURA, The Inflammation-lipocalin2 axis may contribute to the development of chronic kidney disease., Nephrol Dial Transplant, 10.1093/ndt/gft449. , 29, 3, 611-618, 2014.03.
62. Fusanori NISHIMURA, 福田 隆男, Terukazu Sanui, 豊田敬介, 田中麗, Identification of novel amelogenin-binding proteins by proteomics analysis., PLOS One, 10.1371/journal.pone.0078129., 8, 10, e78129, 2013.10.
63. Fusanori NISHIMURA, Par14 associates with IRS-1, thereby enhancing insulin-induced IRS-1 phosphorylation and metabolic actions., J Biol Chem, 10.1074/jbc.M113.485730., 288, 28, 20692-20701, 2013.07.
64. Fusanori NISHIMURA, Resistin-Like Molecule β Is Abundantly Expressed in Foam Cells and Is Involved in Atherosclerosis Development., Arterioscler Thromb Vasc Biol, 10.1161/ATVBAHA.113.301546., 33, 8, 1986-1993, 2013.08.
65. Fusanori NISHIMURA, Integrator complex plays an essential role in adipose differentiation., Biochem Biophys Res Commu, 10.1016/j.bbrc.2013.03.029., 434, 2, 197-202, 2013.05.
66. Fusanori NISHIMURA, RASSF6 expression in adipocytes is down-regulated by interaction with macrophages., PLOS One, 10.1371/journal.pone.0061931. , 8, 4, e61931, 2013.04.
67. Fusanori NISHIMURA, Circulating TNF receptor 2 is associated with the development of chronic kidney disease in non-obese Japanese patients with type 2 diabetes., Diabetes Res Clin Pract, 99, 2, 145-150, 2013.02.
68. Fusanori NISHIMURA, Pain-releasing action of Platelet-activating factor (PAF) antagonists in neuropathic pain animal models and the mechanisms of action., Eur J Pain, 17, 8, 1156-1167, 2013.09.
69. Fusanori NISHIMURA, Flavonol-containing phosphorylated pullulan may attenuate pulp inflammation, INTERNATIONAL ENDODONTIC JOURNAL, 10.1111/j.1365-2591.2012.02095.x, 46, 2, 119-127, 2013.02.
70. Fusanori NISHIMURA, Establishment of an ex vivo pulpitis model by co-culturing immortalized dental pulp cells and macrophages, INTERNATIONAL ENDODONTIC JOURNAL, 10.1111/j.1365-2591.2012.02074.x, 45, 12, 1103-1108, 2012.12.
71. Fusanori NISHIMURA, Improvement of glycated hemoglobin in Japanese subjects with type 2 diabetes by resolution of periodontal inflammation using adjunct topical antibiotics: results from the Hiroshima Study. , Diabetes Res Clin Pract, 100, 53-60, 2013.04.
72. Fusanori NISHIMURA, Dentin sialophosphoprotein and dentin matrix protein-1: Two highly phosphorylated proteins in mineralized tissues, ARCHIVES OF ORAL BIOLOGY, 10.1016/j.archoralbio.2012.03.005, 57, 9, 1165-1175, 2012.09.
73. Fusanori NISHIMURA, Role of Pin1 Protein in the Pathogenesis of Nonalcoholic Steatohepatitis in a Rodent Model, JOURNAL OF BIOLOGICAL CHEMISTRY, 10.1074/jbc.M112.397133, 287, 53, 44526-44535, 2012.12.
74. Fusanori NISHIMURA, Angiotensin receptor 1 blocker valsartan normalizes gene expression profiles of 3T3-L1 adipocytes altered by co-culture with LPS-treated RAW264.7 macrophages, OBESITY RESEARCH & CLINICAL PRACTICE, 10.1016/j.orcp.2012.05.005, 6, 4, E288-E297, 2012.10.