Pulsed Electromagnetic Field (PEMF) Treatment Ameliorates Murine Model of Collagen-Induced Arthritis.

Rheumatoid arthritis (RA) is an autoimmune disease of the joint synovial membranes. RA is difficult to prevent or treat; however, blocking proinflammatory cytokines is a general therapeutic strategy. Pulsed electromagnetic field (PEMF) is reported to alleviate RA's inflammatory response and is being studied as a non-invasive physical therapy. In this current study, PEMF decreased paw inflammation in a collagen-induced arthritis (CIA) murine model. PEMF treatment at 10 Hz was more effective in ameliorating arthritis than at 75 Hz. In the PEMF-treated CIA group, the gross inflammation score and cartilage destruction were lower than in the untreated CIA group. The CIA group treated with PEMF also showed lower serum levels of IL-1ß but not IL-6, IL-17, or TNF-α. Serum levels of total anti-type II collagen IgG and IgG subclasses (IgG1, IgG2a, and IgG2b) remained unchanged. In contrast, tissue protein levels of IL-1ß, IL-6, TNF-α, receptor activator of nuclear factor kappa-Β (RANK), RANK ligand (RANKL), IL-6 receptor (IL-6R), and TNF-α receptor1 (TNFR1) were all lower in the ankle joints of the PEMF-treated CIA group compared with the CIA group. The results of this study suggest that PEMF treatment can preserve joint morphology cartilage and delay the occurrence of CIA. PEMF has potential as an effective adjuvant therapy that can suppress the progression of RA.

Promotion of Colitis in B Cell-Deficient C57BL/6 Mice Infected with Enterotoxigenic Bacteroides fragilis.

Enterotoxigenic Bacteroides fragilis (ETBF) causes colitis and is implicated in inflammatory bowel diseases and colorectal cancer. The ETBF-secreted B. fragilis toxin (BFT) causes cleavage of the adherence junction, the E-cadherin, resulting in the large intestine showing IL-17A inflammation in wild-type (WT) mice. However, intestinal pathology by ETBF infection is not fully understood in B-cell-deficient mice. In this study, ETBF-mediated inflammation was characterized in B-cell-deficient mice (muMT). WT or muMT C57BL/6J mice were orally inoculated with ETBF and examined for intestinal inflammation. The indirect indicators for colitis (loss of body weight and cecum weight, as well as mortality) were increased in muMT mice compared to WT mice. Histopathology and inflammatory genes (Nos2, Il-1ß, Tnf-α, and Cxcl1) were elevated and persisted in the large intestine of muMT mice compared with WT mice during chronic ETBF infection. However, intestinal IL-17A expression was comparable between WT and muMT mice during infection. Consistently, flow cytometry analysis applied to the mesenteric lymph nodes showed a similar Th17 immune response in both WT and muMT mice. Despite elevated ETBF colonization, the ETBF-infected muMT mice showed no histopathology or inflammation in the small intestine. In conclusion, B cells play a protective role in ETBF-induced colitis, and IL-17A inflammation is not attributed to prompted colitis in B-cell-deficient mice. Our data support the fact that B cells are required to ameliorate ETBF infection-induced colitis in the host.

Conditions for CNT - Coated Textile Sensors Applied to Wearable Platforms to Monitor Limb Joint Motion.

Despite recent research on joint motion measurement to monitor human body movement, current measurement techniques and tools have significant limitations, including requiring large space for measurement and causing discomfort in test subjects wearing motion sensors. Our study aims, first, to develop carbon nanotube (CNT)-based textile joint motion sensors. Second, ours study aims to identify the most suitable CNT-based sensor structure and attachment method for use on a wearable platform during general exercise speeds. Lastly, we used these sensors on the human body, using sleeves and legs to find the most stable location, and we used the CNT-based sensor condition to monitor joint motions. We utilized our CNT-based sensor, which has proper elasticity as well as conductivity, and applied it to the elbow and knee joints. Based on the strain gauge principle, we monitored the variance of electric resistance that occurred when the CNT-based sensor was stretched due to limb motion. Our study tested 48 types of sensors. These sensors were applied to the CNT using different base knit textiles as well as different attachment methods, layers, sensor lengths, and sensor widths. The four most successful sensor types, which showed superior efficacy over the others in joint motion measurement, were selected for further study. These four sensors were then used to measure the elbow and knee joint motions of human subjects by placing them on different locations on sleeves and legs. The CNT knit textile sensors best suited to measuring joint motions are those with a double-layered CNT knit and 5 cm long × 0.5 cm or 1 cm wide sensors attached to a polyester¬-based knit using a welding method. The best position for the sensor to more stably monitor joint motions was the "below hinge position" from the elbow or knee hinge joint. Our study suggests an alternative strategy for joint-motion measurement that could contribute to the development of more comfortable and human-friendly methods of human limb motion measurement.

Anti­inflammatory effects of Ciwujianoside C3, extracted from the leaves of Acanthopanax henryi (Oliv.) Harms, on LPS­stimulated RAW 264.7 cells.

The present study aimed to investigate the unknown mechanisms underlying the anti­inflammatory activity of Ciwujianoside C3 (CJS C3), extracted from the leaves of Acanthopanax henryi Harms, on lipopolysaccharide (LPS)­stimulated RAW 264.7 cells. Cells were treated with CJS C3 for 1 h prior to the addition of 200 ng/ml LPS. Cell viability was measured using the MTS assay. Nitric oxide levels were determined by Griess assay. Proinflammatory cytokine production was measured by enzyme­linked immunosorbent assay. The expression levels of cyclooxygenase (COX)­2, inducible nitric oxide synthase (iNOS), and mitogen­activated protein kinases (MAPKs) were investigated by western blotting, reverse transcription (RT)­polymerase chain reaction (PCR) and RT­quantitative PCR. Nuclear factor (NF)­κB/p65 localization, and interaction of the TLR4 receptor with LPS was examined by immunofluorescence assay. The results indicated that CJS C3 exhibited no cytotoxicity at the measured concentrations. Treatment with CJS C3 inhibited NO production, proinflammatory cytokine levels, including interleukin (IL)­6, tumor necrosis factor (TNF)­α, and prostaglandin E2 (PGE2), and protein and mRNA expression levels of iNOS and COX­2. Furthermore, CJS C3 suppressed phosphorylation of extracellular signal­regulated kinases and c­jun N­terminal kinases. It was also able to suppress activation of NF­κB via inhibition of the TLR4 signaling pathway. These results suggested that CJS C3 exerts inhibitory effects on LPS­induced PGE2, NO, IL­6 and TNF­α production. In addition, iNOS and COX­2 expression was decreased in murine macrophages. These inhibitory effects may be achieved via suppression of MAPKs and NF­κB phosphorylation following inhibition of the TLR4 signaling pathway.

In vitro Potential Effect of Morin in the Combination with ß-Lactam Antibiotics Against Methicillin-Resistant Staphylococcus aureus.

Morin, a plant-derived flavonol, is known to be an effective inhibitor of Gram-positive bacteria. In this study, we explored the combined effect of morin with ß-lactam antibiotics against methicillin-resistant Staphylococcus aureus (MRSA), a multidrug-resistant pathogen. The anti-MRSA activity of morin was investigated by the broth microdilution method, checkerboard dilution test, and time-kill curve assay. The expression of the resistant protein, penicillin-binding protein (PBP2a) encoded by mecA, was analyzed by the Western blotting method in the presence of morin and oxacillin. An increased susceptibility of MRSA toward oxacillin was observed in the presence of morin. The protein level of PBP2a was reduced when MRSA (ATCC 33591) was treated with the combination of morin and oxacillin, indicating that the combination of morin and oxacillin potentiates the killing effect against MRSA. The present study indicates that the killing effect by the combinative treatment of morin and ß-lactam antibiotic is dependent on the PBP2a-mediated resistance mechanism.

Quercetin prevents adipogenesis by regulation of transcriptional factors and lipases in OP9 cells.

With the industrialization of society, the increase in the prevalence of obesity and metabolic disorders has become an important health concern in a number of countries. Quercetin (3,30,40,5,7-pentahydroxyflavone) is well known as a bioactive flavonoid in a variety of biological resources. The aim of the present study was to explore the machanisms responsible for the anti-adipogenic activity of quercetin and its effects on the lipolysis in OP9 mouse stromal cells which rapidly differentiate into adipocytes. The differentiation of OP9 cells into adipocytes was evaluated by the measurement of lipid accumulation by Oil Red O (ORO) staining; lipid accumulation was significantly impaired by treatment with quercetin. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis were used to measure the expression levels of CCAAT/enhancer binding protein α (C/EBPα), proliferator-activated receptor γ (PPARγ), sterol regulatory element-binding protein-1 (SREBP-1) and fatty acid synthase (FAS). The mRNA expression levels of lipases, such as adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL) and lipoprotein lipase (LPL) were also measured by RT-PCR. Quercetin significantly decreased the expression of transcription factors, including C/EBPα, PPARγ and SREBP-1c both at the protein and mRNA level. The results from the present study demonstrate that quercetin prevents adipogenesis by upregulating ATGL and HSL expression and downregulating FAS, LPL and adipocyte fatty acid-binding protein (aP2) expression, as well as the expression of transcription factors. Our data suggest that quercetin has therapeutic potential by regulating the expression of transcriptional factors and enzymes associated with adipogenesis.

Protective effect of ixerisoside A against UVB-induced pro-inflammatory cytokine production in human keratinocytes.

Human skin is the first line of defense for the protection of the internal organs of the body from different stimuli. Ultraviolet B (UVB), one of the harmful radiations for skin, is widely known to induce abnormally increased cytokine release from keratinocytes leading to inflammatory skin disorders. IL-6 and IL-8 induce an acute-phase response and stimulate leukocyte infiltration in the skin. Previous studies have shown that chronic exposure to UVB radiation increases cyclooxygenase-2 (COX­2) expression through various cell signaling pathways, resulting in skin cancer. Recent studies have shown that the activation of extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 MAPK is strongly correlated with acute inflammation and development of skin cancer caused by an increased expression of COX-2. Ixerisoside A (IXA) is an active constituent of Ixeris dentata of the Compositae (Asteraceae) family. The effect of IXA on skin inflammation has yet to be elucidated. To determine the anti-inflammatory effects of IXA, we examined its effect on UVB-induced pro-inflammatory cytokine production in human keratinocytes (HaCaT cells) by observing these cells in the presence or absence of IXA. In this study, pro-inflammatory cytokine production was determined by enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (rt-pcr), and western blot analysis to evaluate the activation of mitogen-activated protein kinases (MAPKs). IXA inhibited UVB-induced production of the pro-inflammatory cytokines IL-6 and IL-8 in a dose-dependent manner. Moreover, IXA inhibited the expression of COX-2, ERK, JNK, and p38 MAPKs, indicating that the secretion of the pro-inflammatory cytokines IL-6 and IL-8, and COX-2 expression was inhibited by blocking MAPK phosphorylation. These results indicated that IXA potentially protects against UVB-induced skin inflammation.

Synergistic effects of oxyresveratrol in conjunction with antibiotics against methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) infection is a serious clinical problem worldwide. The aim of the present study was to examine the antimicrobial activity of oxyresveratrol (ORV) against MRSA. The antimicrobial activity of ORV was evaluated against three strains of MRSA and one methicillin-susceptible S. aureus (MSSA) strain using a minimal inhibitory concentration (MIC) assay, MTT colorimetric assay, checkerboard dilution test and time-kill assay. The MIC of ORV for all strains was moderate at 125 µg/ml. Of note, the antimicrobial activity and fractional inhibitory concentration index values of ORV were markedly increased in the presence of a non-growth inhibitory dose of certain antibiotics. Time-kill curves revealed that a combination of ORV with ciprofloxacin or with gentamicin reduced bacterial counts to below the lowest detectable limit after 24 h. These effective combinations may be used as potential antimicrobial regimens for use in the management of MRSA.

Neuroprotective effect of undecylenic acid extracted from Ricinus communis L. through inhibition of µ-calpain.

The key neuropathological features of Alzheimer's disease are abnormal deposition of Aß plaques and insoluble Aß peptides in extracellular brain and intracellular neurofibril tangles induced by abnormal tau hyperphosphorylation. µ-Calpain is one of the factors that bridge these Aß- and hyperphosphorylated tau-mediated pathological pathways. Undecylenic acid (UDA), a naturally occurring unsaturated fatty acid, was discovered as a µ-calpain inhibitor by screening a chemical library using a substrate specific µ-calpain assay method. UDA inhibited Aß oligomerization and Aß fibrillation and reversed Aß-induced neuronal cell death. In addition, UDA scavenged ROS and reversed the levels of proapoptotic proteins induced by ROS in SH-SY5Y cells. UDA inhibited µ-calpain activity with better potency than the known peptide-like µ-calpain inhibitor, MDL28170, in SH-SY5Y and HEK293T cells transfected with the catalytic subunit of µ-calpain. These results suggest that UDA is a novel non-peptide-like µ-calpain inhibitor with good cell permeability and potent neuroprotective effect.

Heart monitoring garments using textile electrodes for healthcare applications.

We measured the electrical activity signals of the heart through vital signs monitoring garments that have textile electrodes in conductive yarns while the subject is in stable and dynamic motion conditions. To measure the electrical activity signals of the heart during daily activities, four types of monitoring garment were proposed. Two experiments were carried out as follows: the first experiment sought to discover which garment led to the least displacement of the textile electrode from its originally intended location on the wearer's body. In the second, we measured and compared the electrical activity signals of the heart between the wearer's stable and dynamic motion states. The results indicated that the most appropriate type of garment sensing-wise was the "cross-type", and it seems to stabilize the electrode's position more effectively. The value of SNR of ECG signals for the "cross-type" garment is the highest. Compared to the "chest-belt-type" garment, which has already been marketed commercially, the "cross-type" garment was more efficient and suitable for heart activity monitoring.

Synthesis and pharmacological evaluation of new methyloxiranylmethoxyxanthone analogues.

In order to develop potential anti-cancer agents that act on topoisomerase II and DNA, we have synthesized 12 new xanthone derivatives. In the cytotoxicity test, compounds 17 and 31 exhibited 2- to 7-fold stronger inhibitory activity than adriamycin against most cancer cell lines tested. Halohydrin group-tethered compounds 19, 21 and 27 showed comparable topoisomerase II inhibitory activity to etoposide at 100 microM concentration. In the DNA cross-linking test, compounds 20, 30 and 31 produced DNA cross-linked adducts and compound 30 was the strongest DNA cross-linker. Based on the combined pharmacological results, we suspected that the strong anti-cancer activity of compounds 16, 17, 20, 30 and 31 originated from the DNA mono-alkylation or cross-linking properties of the compounds.

Identification of 3-acetyl-2-aminoquinolin-4-one as a novel, nonpeptidic scaffold for specific calpain inhibitory activity.

A series of 3-acetyl-2-aminoquinolin-4-one derivatives selected from the Korean Chemical Bank were screened for calpain inhibitory activity by using a high-throughput fluorimetric calpain assay. We identified a potent and selective mu-calpain inhibitor, compound 17, whose specificity and efficacy for mu-calpain inhibition was better than MDL28170. Docking studies revealed that the efficacy of its inhibitory effect on calpain depended on the size and charge properties of the substitutions on the phenylamino ring.

New insight for fluoroquinophenoxazine derivatives as possibly new potent topoisomerase I inhibitor.

Fluoroquinolones, represented by ciproxacin and norfloxacin, are well-known clinical antimicrobial agents, and their phenyl ring expanded quinophenoxazines are reported as possible antitumor active compounds. These quinophenoxazines are known to inhibit DNA topoisomerase II essential for cell replication cycle. But there were no reports for topoisomerase I inhibition study for these compounds. In this report, we have prepared a few quinophenoxazine analogues and tested their topoisomerases I and II inhibitory activities and cytotoxicity. From the result, we found that quinophenoxazine analogues possessed strong topoisomerase I inhibitory capacity as well as topoisomerase II inhibition. Among the compounds prepared, A-62176 analogues showed strong topoisomerases I and II inhibitory activities. Interestingly, compound 8 missing the 3-aminopyrrolidine moiety at C2 position has similar potent inhibitory capacity against topoisomerases I and II at higher concentrations (20 and 10 microM, respectively). But compound 8 inhibited topoisomerase I function more selectively at lower concentration, 2 microM. Our observation might strongly implicate that fluoroquinophenoxazines can be developed as efficient topoisomerase I inhibitor with the elaborate modification.