STAT3 phosphorylation inhibition for treating inflammation and new bone formation in ankylosing spondylitis.

OBJECTIVE: AS is a rheumatic disease characterized by chronic inflammation and bony ankylosis. This study was to evaluate whether a signal transducer and activator of transcription 3 phosphorylation inhibitor (stat3-p Inh) could treat both chronic inflammation and bone formation in AS. METHODS: Primary AS osteoprogenitor cells and spinal entheseal cells were examined for osteogenic differentiation. SF mononuclear cells (SFMCs) and lamina propria mononuclear cells (LPMCs) were obtained from AS patients. Inflammatory cytokine-producing cells were analysed using flow cytometry and ELISA. Female SKG mice were treated with stat3-p Inh, IL-17A blocker or vehicle. Inflammation and new bone formation were evaluated using immunohistochemistry, PET and micro-CT. RESULTS: In the SKG mouse model, stat3-p Inh significantly suppressed arthritis, enthesitis, spondylitis and ileitis. In experiments culturing SFMCs and LPMCs, the frequencies of IFN-γ-, IL-17A- and TNF-α-producing cells were significantly decreased after stat3-p Inh treatment. When comparing current treatments for AS, stat3-p Inh showed a comparable suppression effect on osteogenesis to Janus kinase inhibitor or IL-17A blocker in AS-osteoprogenitor cells. Stat3-p Inh suppressed differentiation and mineralization of AS-osteoprogenitor cells and entheseal cells toward osteoblasts. Micro-CT analysis of hind paws revealed less new bone formation in stat3-p Inh-treated mice than vehicle-treated mice (P = 0.005). Hind paw and spinal new bone formation were similar between stat3-p Inh- and anti-IL-17A-treated SKG mice (P = 0.874 and P = 0.117, respectively). CONCLUSION: Stat-3p inhibition is a promising treatment for both inflammation and new bone formation in AS.

Crystal structure of the indole-3-acetic acid-catabolizing enzyme DAO1 from Arabidopsis thaliana.

Indole-3-acetic acid (IAA), the major form of the plant hormone auxin, regulates almost every aspect of plant growth and development. Therefore, auxin homeostasis is an essential process in plants. Different metabolic routes are involved in auxin homeostasis, but the catabolic pathway has remained elusive until recent studies identified DIOXYGENASE FOR AUXIN OXIDATION (DAO) from rice and Arabidopsis thaliana. DAO, a member of the 2-oxoglutarate/Fe(II)-dependent oxygenase (2ODO) family, constitutes a major enzyme for IAA catabolism. This enzyme catalyzes, with the cosubstrate 2-oxoglutarate, the conversion of IAA into 2-oxoindole-3-acetic acid, a functionally inactive oxidative product of IAA. Here, we report a crystal structure of the unliganded DAO1 from A. thaliana (AtDAO1) and its complex with 2-oxoglutarate. AtDAO1 is structurally homologous with members of the 2ODO family but exhibits unique features in the prime substrate IAA binding site. We provide structural analyses of a putative binding site for IAA, supporting possible structural determinants for the substrate specificity of AtDAO1 toward IAA.

Factors Affecting Maternal and Fetal Outcomes of Non-Obstetric Surgery and Anesthesia during Pregnancy: a Retrospective Review of Data at a Single Tertiary University Hospital.

BACKGROUND: Anesthesia during pregnancy for non-obstetric surgery is generally known to have a negative impact on maternal and fetal outcomes. We assessed the risk of adverse outcomes in fetuses and mothers associated with non-obstetric surgery. METHODS: This retrospective study analyzed clinical data on pregnant women who received non-obstetric surgeries at a tertiary university hospital. We reviewed maternity admissions using hospital administrative data during the last 16 years. The outcome assessment included the presence of preterm labor, premature birth, abortion, or stillbirth and the data of newborns. Statistical analyses were performed using the t-test, χ² test, and multiple logistic regression was used for risk analysis. RESULTS: The incidence of non-obstetric surgery during pregnancy was 0.96%. Gestational age at or above 20 weeks increased the risk of all adverse events 4.5 fold when it was compared to gestational age less than 20 weeks, although the events were only preterm labor or premature birth and no fetal loss. All fetal loss cases occurred in patients at less than 20 weeks of pregnancy. The risk of adverse outcome increased by 2% for every 1 minute increase in anesthesia time. Babies of the mothers who had the adverse outcome event showed lower birth weight and higher neonatal intensive care unit admission rate than those of babies of the mothers without any adverse event after the surgery. CONCLUSION: Physicians should acknowledge and prepare for common possible adverse events at the stage of pregnancy after non-obstetric surgery, and effort to shorten the duration of surgery and anesthesia is needed.

Resveratrol inhibits LXRα-dependent hepatic lipogenesis through novel antioxidant Sestrin2 gene induction.

Liver X receptor-α (LXRα), a member of the nuclear receptor superfamily of ligand-activated transcription factors, regulates de novo fatty acid synthesis that leads to stimulate hepatic steatosis. Although, resveratrol has beneficial effects on metabolic disease, it is not known whether resveratrol affects LXRα-dependent lipogenic gene expression. This study investigated the effect of resveratrol in LXRα-mediated lipogenesis and the underlying molecular mechanism. Resveratrol inhibited the ability of LXRα to activate sterol regulatory element binding protein-1c (SREBP-1c) and thereby inhibited target gene expression in hepatocytes. Moreover, resveratrol decreased LXRα-RXRα DNA binding activity and LXRE-luciferase transactivation. Resveratrol is known to activate Sirtuin 1 (Sirt1) and AMP-activated protein kinase (AMPK), although its precise mechanism of action remains controversial. We found that the ability of resveratrol to repress T0901317-induced SREBP-1c expression was not dependent on AMPK and Sirt1. It is well established that hepatic steatosis is associated with antioxidant and redox signaling. Our data showing that expression of Sestrin2 (Sesn2), which is a novel antioxidant gene, was significantly down-regulated in the livers of high-fat diet-fed mice. Moreover, resveratrol up-regulated Sesn2 expression, but not Sesn1 and Sesn3. Sesn2 overexpression repressed LXRα-activated SREBP-1c expression and LXRE-luciferase activity. Finally, Sesn2 knockdown using siRNA abolished the effect of resveratrol in LXRα-induced FAS luciferase gene transactivation. We conclude that resveratrol affects Sesn2 gene induction and contributes to the inhibition of LXRα-mediated hepatic lipogenesis.

Serum miR-3620-3p as a Novel Biomarker for Ankylosing Spondylitis.

Objective: Using microRNA (miR) as a biomarker has been a new way for diagnosing many diseases Although many studies on miR-biomarker have been published, researches on miR-biomarker in ankylosing spondylitis (AS) are limited Therefore, the objective of this study was to valiate a candidate serum miR as a novel disease-specific novel miR for AS. Methods: Total RNAs were extracted from sera samples of patients with AS (n=57), patients with rheumatoid arthritis (RA) (n=37), or healthy controls (HC) (n=19) Through serum miR screening by microarray, differential levels of miR were subsequently validated by real time PCR At the time of serum sampling, clinical values such as sex, age, disease duration, AS-disease activity score, uveitis, peripheral arthritis, enthesitis, human leukocyte antigen-B27 presence, and recent medication were evaluated. Results: We found that the expression level of serum miR-3620-3p in AS was notably lower than that in RA or HC The receiver-operator characteristics curve for determining the diagnostic accuracy showed an area under the curve of 0919 (p<0001) with a sensitivity of 871% and a specificity of 860% Correlation studies showed that the expression level of miR-3620-3p was only associated with the development of uveitis (p<005). Conclusion: Serum miR-3620-3p can be as a new biomarker for diagnosing AS.

Clonorchis sinensis-Derived Protein Attenuates Inflammation and New Bone Formation in Ankylosing Spondylitis.

Helminth infections and their components have been shown to have the potential to modulate and attenuate immune responses. The objective of this study was to evaluate the potential protective effects of Clonorchis sinensis-derived protein (CSp) on ankylosing spondylitis (AS). Cytotoxicity of CSp at different doses was assessed by MTS and flow cytometry before performing experiments. Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were obtained from AS patients. Inflammatory cytokine-producing cells were analyzed using flow cytometry. The levels of INF- γ , IL-17A, TNF-α, and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). SKG mice were treated with CSp or vehicles. Inflammation and new bone formation were evaluated using immunohistochemistry, positron emission tomography (PET), and micro-computed tomography (CT). Treatment with CSp resulted in no reduced cell viability of PBMCs or SFMCs until 24 h. In experiments culturing PBMCs and SFMCs, the frequencies of IFN- γ and IL-17A producing cells were significantly reduced after CSp treatment. In the SKG mouse model, CSp treatment significantly suppressed arthritis, enthesitis, and enteritis. Micro-CT analysis of hind paw revealed reduced new bone formation in CSp-treated mice than in vehicle-treated mice. We provide the first evidence demonstrating that CSp can ameliorate clinical signs and cytokine derangements in AS. In addition, such CSp treatment could reduce the new bone formation of AS.

Effects of various methods of dexmedetomidine administration for sedation in elderly patients undergoing spinal anesthesia: a randomized controlled study.

BACKGROUND: The purpose of this study was to investigate the degree of sedation and the incidence of adverse effects resulting from various methods of administering the initial dose followed by continuous infusion of dexmedetomidine (DEX) for sedation in elderly patients undergoing spinal anesthesia. METHODS: In total, 72 patients aged over 65 years who were to be administered spinal anesthesia were randomly allocated into three groups. The initial doses were injected to the groups as follows: group DD, DEX 0.5 µg/kg for 10 min; group MD, midazolam 0.02 mg/kg; and group D, no initial dose. This was followed immediately by infusing a maintenance dose of DEX 0.5 µg/kg/h to all groups. RESULTS: The Bispectral index (BIS) in the D group was significantly higher than in the other two groups. There were no significant differences in the Ramsay sedation scale (RSS) among the groups. The RSS 3 level was reached in 10 min from the start of sedation in MD and DD groups and in 20 min from the start of sedation in D group. Neither bradycardia nor hypotension was observed in any of the groups. CONCLUSIONS: Patients in all three groups reached the RSS 3 sedating-effect level. However, the group that received continuous infusion only without the initial dose showed higher BIS than the other two groups and reached the RSS 3 later. No adverse events were observed in any of the groups.

Radiographic Progression in Patients With Ankylosing Spondylitis According to Uveitis Based on the Observation Study of Korean Spondyloarthropathy Registry.

OBJECTIVES: This study aims to investigate radiographic progression according to the presence or absence of uveitis in patients with ankylosing spondylitis (AS). PATIENTS AND METHODS: A total of 598 patients (529 males, 69 females, mean age 38.1±9.2 years; range, 18 to 73 years) from the Observation Study of Korean Spondyloarthropathy Registry who met the modified New York criteria for AS were included in this study. At baseline, all data were stratified into two groups according to the presence or absence of uveitis. Baseline and radiographic progression were assessed for five years in this registry. Modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) was read by two radiologists. Reliability was assessed using inter- and intra- class correlation coefficient for each radiograph. Comparison of mSASSS changes was analyzed by analysis of covariance model after adjusting for confounding factors. RESULTS: The evaluation of mSASSS showed good agreement between the two readers. A total of 193 patients (32.27%) had a history of uveitis that presented at a mean age of 39.6 years, including 30 females (15.54%). There were statistically significant differences in age (p=0.01), sex (p=0.04), hip joint involvement (p<0.01), and human leukocyte antigen B27 carrier state (p=0.02) between the two groups according to uveitis. A simple comparison revealed no significant difference in mSASSS change for five years between the two groups (mean: 3.05±0.62 vs. 3.78±0.78, p=0.47). After adjusting for confounding factors in multiple comparisons by Bonferroni correction, patients with uveitis had no significant association with mSASSS change for five years (mean: 6.29±1.32 vs. 5.49±1.39, p=0.68). CONCLUSION: Our study confirms that there is no significant association between uveitis and radiographic progression in patients with AS after adjusting for confounding factors.

Serum miR-214 as a novel biomarker for ankylosing spondylitis.

OBJECTIVE: Serum microRNA (miR) in ankylosing spondylitis (AS) patients has been rarely identified. The objective of this study was to find AS-specific miR in sera of patients with AS. METHODS: Total RNAs were isolated from whole sera of patients with AS, patients with rheumatoid arthritis (RA), and healthy controls (HC) using miRNeasy Serum/Plasma Kit. The presence of miR was assayed using Agilent 2100 Bioanalyzer Small RNA assay. Each RNA sample was used for miR microarray. To verify microarray results, candidate circulating miRs were validated by quantitative polymerase chain reaction (qPCR) using samples from patients with AS (n = 65), patients with RA (n = 25), and HCs (n = 39). Cycle threshold values were converted to copy numbers by drawing a standard curve using a synthetic chemical standard. All clinical values were also evaluated at the time of miR isolation. RESULTS: A total of 887 miRs were screened for three groups. Lower expression of miR-214 in AS than in HC and RA was observed after normalization of raw data. Finally, lower expression of serum miR-214 was confirmed in AS after validation by qPCR. Correlation analysis showed that the level of miR-214 of AS was significantly associated with Ankylosing Spondylitis Disease Activity Score-C-reactive protein (r = 0.299, P = 0.02). However, other disease-specific variables showed no statistical significance: gender (P = 0.286), peripheral arthritis (P = 0.634), enthesitis (P = 0.464), dacylitis (P = 0.750), psoriasis (P = 0.552), inflammatory bowel disease (P = 0.369), human leukocyte antigen-B27 positivity (P = 0.473), use of non-steroidal anti-inflammatory drugs (P = 0.448), and use of tumor necrosis factor-blocker in the last 3 months (P = 0.505). CONCLUSION: miR-214 may serve as a noninvasive biomarker for diagnosis of AS. In addition, expression level of miR-214 was associated with disease activity.

O-Methylated flavonol isorhamnetin prevents acute inflammation through blocking of NF-κB activation.

Here, we isolated isorhamnetin, a natural 3'-O-methylated flavonoid, from water dropwort (Oenanthe javanica, Umbelliferae) and investigated its ability to protect against acute inflammation in vivo and in vitro. To induce paw swelling, the hind paw of each rat was injected with a carrageenan 1h after vehicle or isorhamnetin treatment. In vitro effect and mechanism studies were performed in lipopolysaccharide (LPS)-activated macrophages. Administration of isorhamnetin markedly inhibited the swelling volume and the thickness of hind paws. Moreover, isorhamnetin significantly reduced inflammatory cell infiltration and pro-inflammatory gene expression in rats. Isorhamnetin pretreatment inhibited inducible nitric oxide synthase (iNOS) expression and NO release in LPS-stimulated cells. Activation of nuclear factor-kappa B (NF-κB) and activating protein-1 (AP-1) is the key step in the iNOS gene induction. Isorhamnetin specifically inhibited NF-κB luciferase activity, but not AP-1. Pretreatment with isorhamnetin suppressed NF-κB nuclear translocation in accordance with decreased phosphorylation and degradation of inhibitory-κB. Consistently, TNF-α, IL-1ß and IL-6 expression, representative NF-κB target genes, were almost completely prohibited by isorhamnetin. Furthermore, isorhamnetin inhibited LPS-induced JNK and AKT/IKKα/ß phosphorylation. Our results suggest that isorhamnetin inhibited JNK, and AKT/IKKα/ß activation, leading to NF-κB inactivation, which might contribute to the inhibition of the acute inflammatory response.

Nrf2-ARE pathway regulates induction of Sestrin-2 expression.

The Sestrin2 (Sesn2) gene encodes a conserved antioxidant protein that is induced on oxidative stress and protects cells against reactive oxygen species. NF-E2-related factor-2 (Nrf2) is an essential transcription factor that regulates expression of a variety of antioxidant genes via binding to the antioxidant-response element (ARE), but the role of Nrf2 in Sesn2 gene expression has not been elucidated yet. The present study investigated whether the Nrf2-ARE pathway regulates Sesn2 gene expression and the identification of the molecular mechanism. The Nrf2 activators, tert-butylhydroquinone (t-BHQ) and sulforaphane (SFN), up-regulated Sesn2 expression in a dose- and time-dependent manner in hepatocytes. Also, t-BHQ increased Sesn2 mRNA and luciferase gene activity, whereas the levels of Sesn1 and Sesn3 mRNA were not affected by t-BHQ treatment. The specific role of Nrf2 in Sesn2 induction was verified by using Nrf2 overexpression plasmid and Nrf2 knockout or knockdown cells. In silico analysis of the 5' upstream region of Sesn2 gene identified a putative ARE sequence. Deletion of the putative ARE demonstrated that the ARE from -550 to -539 bp in the human Sesn2 promoter was critical for the Nrf2-mediated response. Moreover, SFN injection increased Sesn2 mRNA and protein levels in the livers of mice. Knockdown experiments with Sesn2 siRNA showed that Sesn2 is required for the Nrf2-mediated cytoprotective activity against hydrogen peroxide. Our results suggest that the Nrf2-ARE pathway is critical for Sesn2 gene expression and might protect against oxidative stress.