AGEs increase lipocalin 2 expression

Background and Objectives: Diabetes mellitus (DM), a risk factor of periodontal diseases, exacerbates the pathological condition of periodontitis. A major factor for DM complications is advanced glycation end-products (AGEs) that accumulate in periodontal tissues and cause inflammatory events. Lipocalin 2 (LCN2) is an antimicrobial peptide and inflammation-related factor, and LCN2 levels increase in DM. In the present study, the effects of AGEs and lipopolysaccharide of Porphyromonas gingivalis (P.g-LPS) on LCN2 expression in human oral epithelial cells (TR146 cells) and the role of secreted LCN2 in periodontitis with DM were investigated. Material and Methods: TR146 cells were cultured with AGEs (AGE2) and control BSA and cell viability was estimated, or with P.g-LPS. Conditioned medium and cell lysates were prepared from cultures of epithelial cells and used for western blotting and ELISA to analyze LCN2, RAGE, IL-6, MAPK and NF-κB. RNA was isolated from AGE-treated TR146 cells and differentiated HL-60 (D-HL-60) cells and used for quantitative real-time PCR to examine the expression of LCN2 and interleukin-6 (IL-6) mRNAs. RAGE- and LCN2-siRNAs (siRAGE, siLCN2) were transfected into epithelial cells, and AGE-induced LCN2 expression was investigated. D-HL-60 cells were co-cultured with TR146 cells that were transfected with siLCN2 and treated with AGEs, IL-6 mRNA expression in D-HL-60 cells and cell migration were investigated. Results: AGEs increased the expression levels of LCN2 and IL-6 in oral epithelial cells. siRAGE and a neutralizing antibody for RAGE inhibited AGE-induced LCN2 expression. AGEs stimulated the phosphorylation of ERK, p38 and NF-kB in epithelial cells, and their inhibitors suppressed AGE-induced LCN2 expression. In contrast, P.g-LPS did not show a significant increase on LCN2 level in TR146 cells that expressed toll-like receptor 2. In co-culture experiments, AGE-induced LCN2 inhibited IL-6 mRNA expression in D-HL-60 cells, and LCN2 knockdown in epithelial cells suppressed HL-60 cell migration. Conclusion: These results suggested that AGEs increase LCN2 expression via RAGE, MAPK, and NF-κB signaling pathways in oral epithelial cells, and secreted LCN2 may influence the pathological condition of periodontitis with DM

Wall-yielding properties of cell walls from elongating cucumber hypocotyls in relation to the action of expansin

The wall-yielding properties of cell walls were examined using frozen-thawed and pressed segments (FTPs) obtained from the elongation zones of cucumber hypocotyls with a newly developed programmable creep meter. The rate of wall extension characteristically changed depending on both tension and pH. By treatment of the FTPs with acid, the yield tension (y) was shifted downward and the extensibility (f) was increased. However, the downward shift of y was greatly suppressed and the increase in f was partly inhibited in boiled FTPs. The boiled FTPs reconstituted with expansin fully recovered the acid-induced downward y shift as well as the increase in f. Even under the tension below y, wall extension took place pH dependently. Such extension was markedly slower (low-rate extension) than that under the tension above y (high-rate extension). At a higher concentration (8 M), urea markedly inhibited the creep ascribable to the inhibition of the acid-induced downward y shift and increase in f. Moderate concentrations (2 M) of urea promoted wall creep pH dependently. The promotion was equivalent to a 0.5 decrease in pH. The promotion of creep by 2 M urea was observed in boiled FTPs reconstituted with expansin but not in boiled FTPs. These findings indicated that the acidfacilitated creep was controlled by y as well as f in cucumber cell walls. However, y and f might be inseparable and mutually related parameters because the curve of the stress extension rate (SER) showed a gradual change from the low-rate extension to the high-rate extension. Expansin played a role in pH-dependent regulation of both y and f. The physiological meaning of the pH-dependent regulation of wall creep under different creep tensions is also discussed with reference to a performance chart obtained from the SER curves

High Glucose Induces Severe Periodontitis.

Background/Aims: Diabetic patients are susceptible to severe periodontitis, but the precise mechanism is not fully understood. Aim of this study was to explore the biological pathogenesis of severe periodontitis in diabetic patients focusing on the crosstalk of human gingival fibroblasts (HGFs) and macrophages. Methods: A total of 70 periodontitis patients with or without diabetes mellitus (DM) were enrolled, and the statistical relationships of diabetic conditions to the periodontal inflammatory parameters were examined by cross-sectional study. In in vitro study, HGFs cell line CRL-2014® (ATCC) and differentiated THP-1 macrophages were cultured with normal glucose (NG: 5.5 mM) or high glucose (HG: 25 mM) condition, and treated with indicated inflammatory factors such as calprotectin (CPT), interleukin (IL)-1β and IL-6. To examine the effects of HG on soluble IL-6 receptor (sIL-6R) production in THP-1 macrophages, the supernatants were collected and the sIL-6R levels were measured by ELISA. To examine the effects of HG on IL-1β or IL-6-induced matrix metalloproteinase (MMPs) production in HGFs, the supernatants were collected. Levels of MMP-1 and tissue inhibitor of MMP-1 (TIMP-1) were measured by ELISA. Finally, after conditioned medium (CM) from THP-1 macrophages cultured with NG or HG conditions was collected, HGFs were treated with the CM. The supernatants were collected 24 hours later and the levels of MMP-1 and TIMP-1 were measured. To examine the specific effects of IL-1β contained in CM on MMP-1 and TIMP-1 production in HGFs, IL-1 receptor antagonist (IL-1ra) was used. Results: There were statistical correlation between IL-1β and sIL-6R levels in gingival crevicular fluid (GCF) and HbA1c in periodontitis patients with DM (IL-1β: P=0.035, sIL-6R: P=0.040). HG and CPT significantly induced sIL-6R production in THP-1 macrophages. HG significantly enhanced IL-1β or IL-6/sIL-6R-induced MMP-1 production in HGFs. The increase of MMP-1 by both IL-1β and IL-6/sIL-6R was significantly inhibited by specific ERK or IκB inhibitors. Corresponding to the regulation of MMP-1 production, HG condition increased the phosphorylation of p44/42 MAPK and IκBα in HGFs treated with IL-1β or IL-6/sIL-6R. Finally, MMP-1 production in HGFs cultured with HG increased significantly by CM from THP-1 macrophages cultured with HG. The induction of MMP-1 by the CM from THP-1 macrophages cultured with HG was significantly inhibited by dose dependent of IL-1ra in HGFs cultured with HG. Conclusion: Diabetic conditions such as HG induce IL-1β and sIL-6R production from macrophages in inflammatory periodontal tissues and may exacerbate the periodontitis synergistically via MMP-1 production from HGFs

National survey of the association of depressive symptoms with the number of off duty and on-call, and sleep hours among physicians working in Japanese hospitals: a cross sectional study

<p>Abstract</p> <p>Background</p> <p>Physicians' mental health may be adversely affected by the number of days of work and time spent on-call, and improved by sleep and days-off. The aim of this study was to determine the associations of depressive symptoms with taking days of off duty, hours of sleep, and the number of days of on-call and overnight work among physicians working in Japanese hospitals.</p> <p>Methods</p> <p>A cross-sectional study as a national survey was conducted by mail. The study population was 10,000 randomly selected physicians working in hospitals who were also members of the Japan Medical Association (response rate 40.5%). Self-reported anonymous questionnaire was sent to assess the number of days off-duty, overnight work, and on-calls, and the average number of sleep hours on days not working overnight in the previous one month. Depressive state was determined by the Japanese version of the Quick Inventory of Depressive Symptomatology. Logistic regression analysis was used to explore the associations between depressive symptoms and the studied variables.</p> <p>Results</p> <p>Among the respondents, 8.3% of men and 10.5% of women were determined to be depressed. For both men and women, depressive state was associated with having no off-duty days and averaging less than 5 hours of sleep on days not doing overnight work. Depressive state was positively associated with being on-call more than 5 days per month for men, and more than 8 days per month for women, and was negatively associated with being off-duty more than 8 days per month for men.</p> <p>Conclusion</p> <p>Some physicians need some support to maintain their mental health. Physicians who do not take enough days-off, who reduced sleep hours, and who have certain number of days on-calls may develop depressive symptoms.</p

Níveis séricos de sódio e potássio em pacientes com Málaria causada pelo Plasmodium vivax em uma unidade de referência para diagnóstico e tratamento da doença

A malária continua sendo, uma das principais doenças infecciosas em todo o mundo. O Plasmodium vivax é a principal espécie causadora de malária no Brasil. O objetivo do presente estudo foi investigar os níveis séricos dos eletrólitos sódio e potássio em pacientes com infecção pelo Plasmodium vivax, bem como sua relação com parâmetros clínicos e laboratoriais de potencial gravidade da doença. Trata-se de um estudo epidemiológico descritivo, sendo avaliadas as concentrações séricas dos eletrólitos sódio e potássio. Foram incluídos 153 pacientes de junho de 2010 a junho de 2016. Neste estudo, a redução dos níveis da concentração média de sódio foi observada no grupo de pacientes que apresentava febre no dia da consulta e temperatura axilar acima de 37.5 °C.No entanto, o mesmo não foi observado em relação ao potássio. Dessa forma, podemos concluir que na malária vivax a concentração sérica de sódio é menor nos pacientes que relatam ter tido febre no dia da consulta e naqueles que apresentam temperatura axilar maior que 37.5 °C. Os níveis séricos de potássio não são diferentes entre os pacientes que apresentaram febre. Sendo assim, o presente estudo ressalta a necessidade do monitoramento dos níveis de sódio nos pacientes com malária aguda causada pelo Plasmodium vivax

Incidence of central serous chorioretinopathy (2011–2018): a nationwide population-based cohort study of Japan

国の診療報酬請求データベースを用いて特殊な網膜剥離の発症率を明らかに --NDBオンサイトリサーチセンター（京都）を活用した初の成果--. 京都大学プレスリリース. 2021-07-19.[Aims] The aim of this study was to elucidate the epidemiological background of central serous chorioretinopathy (CSC), including its incidence and treatment pattern. [Methods] This was a population-based longitudinal cohort study using a nationwide health insurance claims database of the Japan Ministry of Health, Labour and Welfare (MHLW). As Japan employs universal health coverage, the database covers more than 95% of claims issued in Japan. We accessed all data stored in the database with permission from the MHLW. We traced all individuals aged 30 years or older and identified individuals with new onset of CSC between January 2011 and December 2018. CSC cases were categorised by age and sex for each year, and incidence rate was calculated. We also identified major treatments for CSC to elucidate the initial treatment pattern. [Results] During the 8-year period, 247 930 incidences of CSC were identified, among which 75.9% were men. The crude incidence rate (per 100 000 person-years) in the general population aged 30 years or older was 34.0 (95% CI 33.9 to 34.2), in men was 54.2 (95% CI 53.9 to 54.4) and in women was 15.7 (95% CI 15.5 to 15.8). The mean age of onset was lower in men than in women (50.5±12.5 years vs 54.7±13.5 years). Most of the patients with newly diagnosed CSC (86.8%) did not receive major treatment. [Conclusions] The current study provides the nationwide population-based evidence to clarify the detailed epidemiology of CSC. These results could help to understand the pathogenesis and mechanisms of CSC in the future

Sideward Peak of Intermediate Mass Fragments in High Energy Proton Induced Reactions

Intermediate mass fragment (IMF) formation in the 12 GeV proton induced reaction on Au target is analyzed by using a combined framework of a transport model (JAM/MF) and a newly developed non-equilibrium percolation (NEP) model. In this model, we can well reproduce the mass distribution of fragments. In addition, the sideward peaked angular distribution would emerge under the condition that the fragment formation time is very short, around 20 fm/c. Within this short time period, the un-heated part of the residual nucleus is kept to have doughnut shape, then the Coulomb repulsion from this shape strengthens the sideward peak of IMF.Comment: 22 pages, Latex, 6 embedded PS figure

Calprotectin/TLR4 induced induced-Cytokine Production

Calprotectin, a heterodimer of S100A8 and S100A9 molecules, is associated with inflammatory diseases such as inflammatory bowel disease. We have reported that calprotectin levels in gingival crevicular fluids of periodontitis patients are significantly higher than in healthy subjects. However, the functions of calprotectin in pathophysiology of periodontitis are still unknown. The aim of this study is to investigate the effects of calprotectin on the productivity of inflammatory cytokines in human gingival fibroblasts(HGFs). The HGFs cell line CRL-2014®(ATCC) were cultured, and total RNAs were collected to examine the expression of TLR2/4 and RAGE mRNA using RT-PCR. After the cells were treated with S100A8, S100A9 and calprotectin, supernatants were collected and the levels of IL-6 and MCP-1 were measured using ELISA methods. To examine the intracellular signals involved in calprotectin-induced cytokine production, several chemical inhibitors were used. Furthermore, after the siRNA-mediated TLR4 down-regulated cells were treated with S100A8, S100A9 and calprotectin, the levels of IL-6 and MCP-1 were also measured. HGFs showed greater expression of TLR4 mRNA, but notTLR2 and RAGE mRNA compared with human oral epithelial cells. Calprotectin increased significantly the production of MCP-1 and IL-6 in HGFs, and the cytokine productions were significantly suppressed in the cells treated with MAPKs, NF-kBand TLR4inhibitors. Furthermore, calprotectin-mediated MCP-1 and IL-6 production were significantly suppressed in TLR4down-regulated cells. Taken together, calprotectin inducesIL-6 and MCP-1 production in HGFs via TLR4 signaling that involves MAPK and NF-kB, resulting in the progression of periodontitis