ジョウヒ サイボウ ニオケル コウキン ペプチド ノ ハツゲン チョウセツ キコウ ノ カイメイ

Epithelial cells express antimicrobial peptides (AMPs) including calprotectin (S100A8/S100A9), β-defensin and lipocalin, which contribute to a host defense reaction in the innate immune system. However, the regulation of AMP expression is not well elucidated. Interleukin1-α (IL-1α), an autonomous cytokine, regulates keratinocyte differentiation and also induces the expression of keratinocyte growth factor (KGF) in fibroblasts. It is reported that KGF stimulates the growth and differentiation of keratinocytes. I confirmed the expression of calprotectin in gingiva and skin by immunohistochemical staining, and determined the effect of IL-1α and KGF on the expression of several AMPs in human keratinocytes cell line (HaCaT) and human fibroblast cell line (NB1RGB), and then clarified the regulatory mechanism of calprotectin expression induced by IL-1α and KGF with the analyses of microarray, Northern blot, RT-PCR and Western blot. Immunohistochemical staining showed that calprotectin expressed in human normal gingiva and skin, and this expression increased in periodontitis gingivae and psoriasis skins. IL-1α increased KGF mRNA expression in NB1RGB, and KGF up-regulated IL-1α mRNA expression in HaCaT. Microarray analysis revealed that IL-1α increased some AMPs, lipocalin 2 (LCN2), S100A8, S100A9 and secretory leukocyte protease inhibitor (SLPI), showing more than 2-fold expressions in HaCaT. RT-PCR and Northern blot analysis revealed that IL-1α increased mRNA expressions of S100A7, S100A8, S100A9, LCN2, SLPI, and β-defensin 2 (hBD-2) in HaCaT. On the other hand, KGF decreased the expression of S100A7, S100A8, and S100A9, and increased LCN2 and SLPI expressions. KGF did not affect hBD-2 expression. When the inhibitors of mitogen-activated protein kinase (MAPK), including p38, c-Jun N-terminal kinase, and extracellular signal-regulated kinase (ERK) were added to HaCaT, p38 inhibitor suppressed the IL-1α-up-regulated S100A8/S100A9 expression and ERK inhibitor suppressed the KGF-down-regulated S100A8/S100A9 expression. These results indicate that some AMP expressions are regulated by IL-1α and KGF in human keratinocytes and that IL-1α up-regulates and KGF down-regulates S100A8/S100A9 genes through MAPK pathway in keratinocytes

Establishment of leptin-Responsive cell lines from adult mouse hypothalamus

Leptin resistance is considered to be the primary cause of obesity. However, the cause of leptin resistance remains incompletely understood, and there is currently no cure for the leptin-resistant state. In order to identify novel drug-target molecules that could overcome leptin resistance, it would be useful to develop in vitro assay systems for evaluating leptin resistance. In this study, we established immortalized adult mouse hypothalamus-derived cell lines, termed adult mouse hypothalamus (AMH) cells, by developing transgenic mice in which SV40 Tag was overexpressed in chromogranin A-positive cells in a tamoxifendependent manner. In order to obtain leptin-responsive clones, we selected clones based on the phosphorylation levels of STAT3 induced by leptin. The selected clones were fairly responsive to leptin in terms of STAT3, ERK, and Akt phosphorylation and induction of c- Fos mRNA induction. Pretreatment with leptin, insulin, and palmitate attenuated the c-Fos mRNA response to leptin, suggesting that certain aspects of leptin resistance might be reconstituted in this cellular model. These cell lines are useful tools for understanding the molecular nature of the signal disturbance in the leptin-resistant state and for identifying potential target molecules for drugs that relieve leptin resistance, although they have drawbacks including de-differentiated nature and lack of long-time stability

ショウガオールはヒト歯肉線維芽細胞において酸化ストレス反応の調節を介してAGEs誘導性のIL-6およびICAM-1産生を抑制する

Advanced glycation end-products (AGEs) cause diabetes mellitus (DM) complications and accumulate more highly in periodontal tissues of patients with periodontitis and DM. AGEs aggravate periodontitis with DM by increasing the expression of inflammation-related factors in periodontal tissues. 6-Shogaol, a major compound in ginger, has anti-inflammatory and anti-oxidative activities. However, the influence of shogaol on DM-associated periodontitis is not well known. In this study, the effects of 6-shogaol on AGEs-induced oxidative and anti-oxidative responses, and IL-6 and ICAM-1 expression in human gingival fibroblasts (HGFs) were investigated. When HGFs were cultured with 6-shogaol and AGEs, the activities of reactive oxygen species (ROS) and antioxidant enzymes (heme oxygenase-1 [HO-1] and NAD(P)H quinone dehydrogenase 1 [NQO1]), and IL-6 and ICAM-1 expressions were investigated. RAGE expression and phosphorylation of MAPKs and NF-κB were examined by western blotting. 6-Shogaol significantly inhibited AGEs-induced ROS activity, and increased HO-1 and NQO1 levels compared with the AGEs-treated cells. The AGEs-stimulated expression levels of receptor of AGE (RAGE), IL-6 and ICAM-1 and the phosphorylation of p38, ERK and p65 were attenuated by 6-shogaol. These results suggested that 6-shogaol inhibits AGEs-induced inflammatory responses by regulating oxidative and anti-oxidative activities and may have protective effects on periodontitis with DM

CHEMO‑RADIATION FOR MYOEPITHELIAL CARCINOMA

Soft tissue myoepithelial carcinoma (MEC) is an extremely rare mesenchymal tumor that has a poor prognosis unless complete surgical resection is achieved. The present study reported a case of a 38‑year‑old woman with a tumor in the left paraspinal region at L2 to L3 with vertebral destruction. MEC was diagnosed based on molecular pathological examination of a biopsy specimen. Because curative surgery was expected to be difficult, a combination of chemotherapy with doxorubicin and ifosfamide and proton beam therapy as local therapy was performed, resulting in long‑term survival for at least 7.8 years. To the best of our knowledge, this is the first case of soft tissue MEC for which classical cytotoxic chemotherapy and proton beam therapy were effective. Although surgical resection with negative margins is the mainstay of treatment for MEC, adequate doxorubicin‑based systemic therapy and high‑dose radiation therapy may be a feasible alternative in patients with unresectable or advanced MEC. Future studies on the relationship between molecular pathological features, including biomarkers, and the selection of therapeutic agents are warranted

β-defensin 2 synthesized by a cell-free protein synthesis system and encapsulated in liposomes inhibits adhesion of Porphyromonas gingivalis to oral epithelial cells

β-defensin 2 (BD-2), an antimicrobial peptide (AMP), is expressed by oral epithelial cells and plays an important role in innate immunity of the oral cavity. Cell-free protein synthesis (CFPS) systems have been studied for the synthesis of various proteins, however, the synthesis of BD-2 by a CFPS system has not been extensively explored. Liposomes have been developed as tools for drug delivery. A delivery of liposome-encapsulated AMP to oral epithelium may be useful to prevent oral infectious diseases. In the present study, we investigated the antimicrobial activity of the BD-2 protein, artificially synthesized using a CFPS system and encapsulated in liposomes. BD-2 protein was artificially synthesized using template DNA and a reconstituted CFPS system and was identified by western blotting. Bilayer liposomes were prepared using 1,2-dioleoyl-sn-glycero-3-phospho-choline and 3-sn-phosphatidylcholine from egg yolk. The artificially synthesized BD-2 was encapsulated in liposomes, collected by ultrafiltration, and detected by western blotting. Human oral epithelial cells were cultured with the liposome-encapsulated BD-2 and the concentration of BD-2 in the cell lysate of the culture with the synthesized BD-2 was higher than that of the control cultures. The antimicrobial activity of the synthesized BD-2 was investigated by an adhesion assay of Porphyromonas gingivalis to oral epithelial cells. The artificially synthesized BD-2 and its liposome significantly inhibited adhesion of P. gingivalis to oral epithelial cells. These results suggest that artificially synthesized BD-2 and liposome-encapsulated BD-2 shows antimicrobial activity and can potentially play a role in oral healthcare for periodontal diseases

Glycated Albumin in Gingival Crevicular Fluid from Patients With Diabetes

Background: Diabetes mellitus (DM) patients have a high prevalence of periodontitis. DM-associated periodontitis (DM-P) is characterized by severe inflammation and tissue destruction. To diagnose DM-P is important for cures of periodontitis and DM. The purpose of this study was to investigate the levels of glycated albumin (GA), a DM marker, and calprotectin, an inflammatory marker, in gingival crevicular fluid (GCF) from patients with periodontitis and DM. Methods: Seventy-eight subjects participated in this study were the patients with DM, chronic periodontitis (CP), DM-P and healthy individuals (H). GCF and blood were collected from four groups. GA and calprotectin in GCF were analyzed using western blotting and ELISA, and their levels were compared among H, DM, CP, and DM-P groups. GA and glycated hemoglobin (HbA1c) in blood were determined, and the correlation between GCF GA level and blood HbA1c or GA level was investigated. ROC analysis for GCF GA level to predict DM was performed. Results: GA was identified in GCF, and its amount and concentration in GCF samples from DM and DM-P were significantly higher than those of non-DM groups (H and CP). Calprotectin amount in GCF from CP and DM-P was significantly higher than that in H and DM groups. GCF GA level was positively correlated to blood HbA1c and GA level. ROC analysis of GCF GA level showed an optimal cut-off value to predict DM. Conclusions: GA showed a high level in GCF from DM patients. GA and calprotectin in GCF may be useful markers to diagnose DM-associated periodontitis

Gan-Lu-Yin (Kanroin), Traditional Chinese Herbal Extracts, Reduces Osteoclast Differentiation In Vitro and Prevents Alveolar Bone Resorption in Rat Experimental Periodontitis

Gan-Lu-Yin (GLY), a traditional Chinese herbal medicine, shows therapeutic effects on periodontitis, but that mechanism is not well known. This study aims to clarify the precise mechanism by investigating the inhibitory effects of GLY extracts on osteoclastogenesis in vitro and on bone resorption in periodontitis in vivo. RAW264.7 cells are cultured with soluble receptor activator of nuclear factor-kappa B (sRANKL) and GLY extracts (0.01–1.0 mg/mL), and stained for tartrate-resistant acid phosphatase (TRAP) to evaluate osteoclast differentiation. Experimental periodontitis is induced by placing a nylon ligature around the second maxillary molar in rats, and rats are administered GLY extracts (60 mg/kg) daily for 20 days. Their maxillae are collected on day 4 and 20, and the levels of alveolar bone resorption and osteoclast differentiation are estimated using micro-computed tomography (CT) and histological analysis, respectively. In RAW264.7 cells, GLY extracts significantly inhibit sRANKL-induced osteoclast differentiation at a concentration of more than 0.05 mg/mL. In experimental periodontitis, administering GLY extracts significantly decreases the number of TRAP-positive osteoclasts in the alveolar bone on day 4, and significantly inhibits the ligature-induced bone resorption on day 20. These results show that GLY extracts suppress bone resorption by inhibiting osteoclast differentiation in experimental periodontitis, suggesting that GLY extracts are potentially useful for oral care in periodontitis

Multimodality imaging of biatrial myxomas in an asymptomatic patient

AbstractMyxomas are located in the left atrium in 75–80% of cases and almost always present with signs and symptoms of a thromboembolic event. Biatrial myxomas are rare, and their incidence is generally less than 2.5% of all myxomas. We herein present a case of biatrial myxomas as an incidental finding by echocardiography where the patient underwent surgery. Echocardiography continues to be the initial imaging modality for intracardiac masses. Cardiac magnetic resonance provides superior tissue characterization, particularly important in differentiating a myxoma from a thrombus. Appropriate use of these non-invasive imaging modalities may lead to a correct diagnosis and good outcome.<Learning objective: In this report we present a rare case of cardiac biatrial myxomas. Multimodality imaging, especially delayed enhancement cardiac magnetic resonance imaging, provided specific findings for the diagnosis.

Sutdies on Galvanic skin responce (GSR) in relation to psychological condition test for the students.

Psychological anxiety trend was studied by Galvanic skin responce (GSR) 60 students, 7 of male and 53 of female, in School of Health Sciences Tokushima University, and the results were compared with the data from State Anxiety Inventory (STAI) test and Self-rating Depression Scale (SDS) test. Compared GSR with STAI, initial responses of GSR showed postive relation to state anxiety in STAI test ; while compared GSR with SDS, both of spontaneous and initial responses showed reversed correlation. Although spontaneous and initial responce in GSR were more remarkably augmented in the female students than male students, these trends might not be caused only by sex difference because the test environment was not same in two groups. Based on these data, GSR is considered to represent objective indicator of psychological anxiety. However further basic data accumulations are needed in regard to some factors affecting the data such as sex of examiners or examinees and test environment