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Various small molecules have been used as functional probes for tissue imaging in medical diagnosis and pharmaceutical drugs for disease treatment. The spatial distribution, target selectivity, and diffusion/excretion kinetics of small molecules in structurally complicated specimens are critical for function. However, robust methods for precisely evaluating these parameters in the brain have been limited. Herein, we report a new method termed “fixation-driven chemical cross-linking of exogenous ligands (FixEL), ” which traps and images exogenously administered molecules of interest (MOIs) in complex tissues. This method relies on protein-MOI interactions and chemical cross-linking of amine-tethered MOI with paraformaldehyde used for perfusion fixation. FixEL is used to obtain images of the distribution of the small molecules, which addresses selective/nonselective binding to proteins, time-dependent localization changes, and diffusion/retention kinetics of MOIs such as the scaffold of PET tracer derivatives or drug-like small molecules., ホルマリン漬けから着想した小分子可視化法 --医薬品開発効率化につながる新たな戦略--. 京都大学プレスリリース. 2022-12-05.

Peucedanum japonicum Thunberg (PJT) has been used in traditional medicine to treat colds, coughs, fevers and other inflammatory diseases. The goal of this study was to investigate whether 3'-isovaleryl-4'-senecioylkhellactone (IVSK) from PJT has anti-inflammatory effects on lung epithelial cells. Anti-inflammatory effects of IVSK was evaluated using phorbol 12-myristate 13-acetate (PMA)-stimulated A549 cells and regular human lung epithelial cells as a reference. IVSK reduced the secretion of inflammatory mediators, interleukin (IL)-8 and monocyte chemoattractant protein-1 (MCP-1) and the mRNA expression of IL-6, IL-8, MCP-1 and IL-1β. Additionally, it inhibited the phosphorylation of IκB kinase (IKK), p65, Iκ-Bα and mitogen-activated protein kinases (MAPKs) p38, JNK, and ERK in A549 cells stimulated with PMA. Moreover, the binding affinity of activator protein-1 (AP-1) and nuclear factor-κB (NF-κB) was significantly reduced in the luciferase assay, and nuclear translocation was markedly inhibited by IVSK in the immunocytochemistry. These findings indicate that IVSK can protect against inflammation through the AP-1 and NF-κB pathway and could possibly be used as a lead compound for the treatment of inflammatory lung diseases.

Objective We investigated whether purpurin inhibits various pathways of inflammation leading to atopic dermatitis. Introduction 1,2,4-Trihydroxyanthraquinone, commonly called purpurin, is an anthraquinone that is a naturally occurring red/yellow dye. Purpurin is a highly antioxidative anthraquinone and previous studies have reported antibacterial, anti-tumor, and anti-oxidation activities in cells and animals. However, the skin inflammatory inhibition activity mechanism study of purpurin has not been elucidated in vitro. Methods In this study, we investigated the anti-inflammatory activity of purpurin in HaCaT (human keratinocyte) cell lines stimulated with a mixture of tumor necrosis factor-alpha (TNF-α)/Interferon-gamma (IFN-γ). The inhibitory effect of Purpurin on cytokines (IL-6, IL-8, and IL-1β) and chemokine (TARC, MDC, and RANTES) was confirmed by ELISA and RT-qPCR. We investigated each signaling pathway and the action of inhibitors through western blots. Results The expression levels of cytokines and chemokines were dose-dependently suppressed by purpurin treatment in TNF-α/IFN-γ-induced HaCaT cells from ELISA and real-time PCR. Purpurin also inhibited protein kinase B (AKT), mitogen-activated protein kinase (MAPKs), and nuclear factor kappa-light-chain-enhancer of activated B (NF-κB) activation in TNF-α/IFN-γ-stimulated HaCaT cells. Additionally, there was a synergistic effect when purpurin and inhibitor were applied together, and inflammation was dramatically reduced. Conclusion Therefore, these results demonstrate that purpurin exhibits anti-inflammatory and anti-atopic dermatitis activity in HaCaT cells.

Various small molecules have been used as functional probes for tissue imaging in medical diagnosis and pharmaceutical drugs for disease treatment. The spatial distribution, target selectivity, and diffusion/extrusion kinetics of small molecules in structurally complicated specimens are critical for function. However, robust methods for precisely evaluating these parameters in the brain have been limited. Herein we report a new method termed “Fixation-driven chemical crosslinking of exogenous ligands (FixEL)” which traps and images exogenously administered molecules-of-interest (MOI) in complex tissues. This method relies on proteins-MOI interactions, and chemical crosslinking of amine-tethered MOI with paraformaldehyde used for perfusion fixation. FixEL is used to obtain images of the distribution of the small molecules and their dynamics, which addresses selective/nonselective binding to proteins, time-dependent localization changes, and diffusion/retention kinetics of MOI such as PET tracer derivatives or drug-like small molecules. Clear imaging of a nanobody distributed in the whole brain was also achieved with high spatial resolution using 2D/3D mode.

We have developed a cell penetrating peptide (CPP) system with high selectivity and penetrability at nanomolar concentrations with a combination of an HER2-selective affibody, ZHER2:342 (ZHER2), and a dimeric α-helical leucine- and lysine-rich peptide, LK-2. ZHER2 and LK-2 are linearly fused together and expressed in a prokaryotic system to create the LK-2-ZHER2 protein, which can successfully distinguish and penetrate HER2-overexpressing cancer cells at nanomolar concentrations. LK-2-ZHER2 has the ability to intracellularly deliver doxorubicin as a conjugate form to enhance its anti-cancer effect on HER2-overexpressing breast cancer cells with a great selectivity. The selective penetrability was confirmed in vitro, in 3D spheroids, and in in vivo models. LK-2-ZHER2 has the capability to overcome the weak points of current CPPs, such as poor penetrability at low concentrations and a lack of selectivity, by combining powerful CPP and affibody sequences.

Methyl p-hydroxycinnamate (MH), an esterified derivative of p-Coumaric acid exerts anti-inflammatory effects on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Based on these effects, the present study investigated the protective role of MH in a mouse model of LPS-induced acute respiratory distress syndrome (ARDS). The results demonstrated that administration of LPS (5 mg/kg intranasally) markedly increased the neutrophil/macrophage numbers and levels of inflammatory molecules (TNF-α, IL-6, IL-1β and reactive oxygen species) in the bronchoalveolar lavage fluid (BALF) of mice. On histological examination, the presence of inflammatory cells was observed in the lungs of mice administered LPS. LPS also notably upregulated the secretion of monocyte chemoattractant protein-1 and protein content in BALF as well as expression of inducible nitric oxide synthase in the lungs of mice; it also caused activation of p38 mitogen-activated protein kinase (MAPK) and NF-κB signaling. However, MH treatment significantly suppressed LPS-induced upregulation of inflammatory cell recruitment, inflammatory molecule levels and p38MAPK/NF-κB activation, and also led to upregulation of heme oxygenase-1 (HO-1) expression in the lungs of mice. In addition, the ability of MH to induce HO-1 expression was confirmed in RAW264.7 macrophages. Taken together, the findings of the present study indicated that MH may exert protective effects against airway inflammation in ARDS mice by inhibiting inflammatory cell recruitment and the production of inflammatory molecules.

3,4,5‑Trihydroxycinnamic acid (THCA) exhibits anti‑inflammatory activity in acute or chronic inflammatory disorders, such as acute lung injury and asthma. The present study investigated the anti‑inflammatory activity of THCA in a tumor necrosis factor‑α/interferon‑γ (TI) mixture‑stimulated human keratinocyte cell line. The results of ELISA and reverse transcription‑quantitative PCR revealed that THCA reduced the secretion and mRNA expression levels of interleukin (IL)‑6; IL‑8; thymus and activation‑regulated chemokine; macrophage‑derived chemokine; regulated upon activation, normal T cell expressed and secreted; and monocyte chemoattractant protein‑1 in TI mixture‑stimulated HaCaT cells. In addition, the results of western blot analysis demonstrated that THCA exerted inhibitory activity on the activation of AKT, ERK and nuclear factor‑κB in TI mixture‑stimulated HaCaT cells. Furthermore, THCA upregulated the expression levels of heme oxygenase‑1 and NAD(P)H:quinone oxidoreductase 1, and the activation of nuclear factor erythroid 2‑related factor 2 in HaCaT cells. These results demonstrated that THCA may exhibit anti‑inflammatory activity in activated HaCaT cells.

The ability to incorporate a desired functionality into proteins of interest in a site-specific manner can provide powerful tools for investigating biological systems and creating therapeutic conjugates. However, there are not any universal methods that can be applied to all proteins, and it is thus important to explore the chemical strategy for protein modification. In this paper, we developed a new reactive peptide tag/probe pair system for site-specific covalent protein labeling. This method relies on the recognition-driven reaction of a peptide tag and a molecular probe, which comprises the lysine-containing short histidine tag (KH6 or H6K) and a binuclear nickel (II)- nitrilotriacetic acid (Ni2+-NTA) complex probe containing a lysine-reactive N-acyl-N-alkyl sulfonamide (NASA) group. The selective interaction of the His-tag and Ni2+-NTA propeles a rapid nucleophilic reaction between a lysine residue of the tag and the electrophilic NASA group of the probe by the proximity effect, resulting in the tag-site-specific functionalization of proteins. We characterized the reactive profile and site-specificity of this method using model peptides and proteins in vitro, and demonstrated the general utility for production of a nanobody-chemical probe conjugate without compromising its binding ability.

A new vehicle is designed for the intracellular delivery of antibodies at nanomolar concentrations by combination of domain Z, a small affibody with strong binding affinity to Fc regions of immunoglobulin G (IgG), and the multimers of LK sequences, α-helical cell penetrating peptides (CPP) with powerful cell penetrating activities. Domain Z and multimeric LK are fused together to form LK-domain Z proteins. The LK-domain Z can bind with IgG at a specific ratio at nanomolar concentrations by simple mixing. The IgG/LK-domain Z complexes can successfully penetrate live cells at nanomolar concentration and the delivery efficiency is strongly dependent upon the concentrations of IgG/LK-domain Z complex as well as the species and subclasses of IgGs. The IgG/LK-domain Z complexes penetrate cells via ATP-dependent endocytosis pathway and the majority of delivered IgG seems to escape endosome to cytosol. Remarkably, the delivered IgGs are able to control the targeted intracellular signaling pathway as shown in the down-regulation of pro-survival genes by the delivery of anti-NF-κB using an LK-domain Z vehicle with a cathepsin B-cleavable linker between the LK sequence and domain Z. The simple but very efficient intracellular delivery method of antibodies at nanomolar concentrations is expected to facilitate profound understanding of cell mechanisms and development of new future therapeutics on the basis of intracellular antibodies.

In this study, we propose a reversible covalent conjugation method for peptides, proteins, and even live cells based on specific recognition between natural amino acid sequences. Two heptad sequences can specifically recognize each other and induce the formation of a disulfide bond between cysteine residues. We show the covalent bond formation and dissociation between peptides and proteins in cell-free conditions and on the surface of live cells. Because heptad sequences consist of natural amino acids, they are expressed in cells without additional preparation and can be used to selectively conjugate peptides, proteins, and cells.

3,4,5-Trihydroxycinnamic acid (THCA) has been reported to possess anti-inflammatory activity. However, the effect of THCA for treating allergic asthma was unknown. Therefore, in the present study, the anti-asthmatic effects of THCA were studied in both in vitro and in vivo studies. In phorbol 12-myristate 13-acetate (PMA)-stimulated A549 airway epithelial cells, THCA pretreatment decreased the mRNA expression and secretion of interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1), and intercellular adhesion molecules 1 (ICAM-1), and reduced the mRNA expression of matrix metalloproteinase 9 (MMP-9). THCA also inhibited PMA-induced protein kinase B (AKT), mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) activation in A549 cells. In lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages, THCA pretreatment suppressed the mRNA expression of ICAM-1 and MMP-9. In addition, THCA suppressed the adhesion of EOL and A549 cells. In ovalbumin (OVA)-administered asthmatic mice, THCA exerted inhibitory activity on IL-5, IL-13, and MCP-1 in bronchoalveolar lavage fluid (BALF) and on OVA-specific immunoglobulin E (IgE) in serum. THCA attenuated the numbers of inflammatory cells in BALF and the influx of inflammatory cell in lung tissues. Furthermore, THCA downregulated the levels of inducible nitric oxide (iNOS), cyclooxygenase-2 (COX-2), and leukotriene B4 (LTB4) expression, mucus production and CREB phosphorylation as well as Penh value. These effects were accompanied by suppression of AKT, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and NF-κB activation. Therefore, the results of the current study suggest that THCA may be a valuable adjuvant or therapeutic in the prevention or treatment of allergic asthma.

Spinal cord microglia contribute to nerve injury‐induced neuropathic pain. We have previously demonstrated that toll‐like receptor 2 (TLR2) signaling is critical for nerve injury‐induced activation of spinal cord microglia, but the responsible endogenous TLR2 agonist has not been identified. Here, we show that nerve injury‐induced upregulation of sialyltransferase St3gal2 in sensory neurons leads to an increase in expression of the sialylated glycosphingolipid, GT1b. GT1b ganglioside is axonally transported to the spinal cord dorsal horn and contributes to characteristics of neuropathic pain such as mechanical and thermal hypersensitivity. Spinal cord GT1b functions as an TLR2 agonist and induces proinflammatory microglia activation and central sensitization. Pharmacological inhibition of GT1b synthesis attenuates nerve injury‐induced spinal cord microglia activation and pain hypersensitivity. Thus, the St3gal2‐GT1b‐TLR2 axis may offer a novel therapeutic target for the treatment of neuropathic pain.

We stapled an amphipathic peptide mainly consisting of leucine (L) and lysine (K) by an azobenzene (Ab) linker for photocontrol of the secondary structure. The cis- trans isomerization of the Ab moieties could stabilize and destabilize the α-helical conformation of the LK peptide along with dramatic change of associated peptide structures in a reversible manner by UV-vis irradiation. The cell-penetrating activities of the LK peptide can be readily regulated by the photocontrol, as the stabilized cis-Ab-LK peptide showed remarkable increase of cell penetration compared to the destabilized trans-Ab-LK peptide. The photoswitchable cell-penetrating peptides would provide important structural information for cell permeability as well as an effective targeting strategy for peptide-based pharmaceuticals with spatiotemporal specificity.

OBJECTIVE In this study, to measure blood pressure (BP) on the basis of human hearing threshold, we proposed a method that detects the audible or inaudible Korotkoff sounds (K-sounds) using the equal loudness contour and automatically assesses the BP. METHODS In this study, we detected the systolic period of K-sounds using cuff pressure oscillation and then converted the K-sounds corresponding to the systolic interval into sound pressure levels (SPLs). Next, the systolic blood pressure (SBP) and diastolic blood pressure (DBP) were assessed by mapping the K-sounds, which were converted into SPLs on an equal loudness contour. RESULTS To validate the accuracy of our proposed method, we compared it with the auscultatory method. The mean differences (mean±SD) in the SBP and DBP were 0.31±1.95 and 1.20±2.17 mmHg, respectively. For the SBP, the linear regression equation was y=0.98x+1.56 mmHg (where x and y represent the auscultatory and the proposed method, respectively), with a SE of estimate of 1.93 mmHg and a correlation coefficient of 0.99. For the DBP, the linear regression equation was y=1.01x-1.94 mmHg, with an SE of estimate of 2.18 mmHg and a correlation coefficient of 0.98. All P values were less than 0.0001 for both regressions. CONCLUSION The auscultatory method of BP monitoring is sensitive to the observer's condition or environmental noise. To overcome these disadvantages, we used the human hearing threshold for objective SBP and DBP automatic assessment, and this method can be applicable to an automatic auscultatory method.

BACKGROUND There have been few recent reports on the methodological quality of meta-analysis, despite the enormous number of studies using meta-analytic techniques in the field of anesthesia. The purpose of this study was to evaluate the quality of meta-analyses and systematic reviews according to the Assessment of Multiple Systematic Reviews (AMSTAR) and Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines in the anesthesia literature. METHODS A search was conducted to identify all meta-analyses ever been published in the British Journal of Anaesthesia (BJA), Anaesthesia, and Korean Journal of Anesthesiology (KJA) between Jan. 01, 2004 and Nov. 31, 2016. We aimed to apply the AMSTAR and PRISMA checklists to all published meta-analyses. RESULTS We identified 121 meta-analyses in the anesthesia literature from January 2004 through the end of November 2016 (BJA; 75, Anaesthesia; 43, KJA; 3). The number of studies published and percentage of 'Yes' responses for meta-analysis articles published after the year 2010 was significantly increased compared to that of studies published before the year 2009 (P = 0.014 for Anaesthesia). In the anesthesia literature as a whole, participation of statisticians as authors statistically improved average scores of PRISMA items (P = 0.004) especially in the BJA (P = 0.003). CONCLUSIONS Even though there is little variability in the reporting and methodology of meta-analysis in the anesthesia literature, significant quality improvement in the reporting was observed in the Anaesthesia by applying the PRISMA checklist. Participation of a statistician as an author improved the reporting quality of the meta-analysis.

Study objective Administration of intranasal dexmedetomidine for sedation is comfortable and effective in children who are afraid of needles, and it offers efficient sedation similar to that of intravenous administration. We performed a systematic review and meta-analysis to evaluate the clinical effects of the pre-procedural administration of intranasal dexmedetomidine. Design We identified randomized controlled trials (RCTs) that compared intranasal dexmedetomidine administration to other administration methods of various sedatives or placebo from MEDLINE, EMBASE, Cochrane, KoreaMed and hand searches of trial registries. Setting Pediatrics who underwent interventional procedures and surgeries. Patients Children under the age of 18. Interventions Studies were included if they were compatible with the criteria that dexmedetomidine was administered intranasally. Measurements We pooled data on the sedation status as the primary outcome and considered the behavioral score, blood pressure, heart rate and side effects to be secondary outcomes. Risk ratio (RR) and the standardized mean difference (SMD) with 95% confidence intervals (CIs) were calculated for dichotomous and continuous outcomes, respectively. Main results This meta-analysis included 11 RCTs. The SMD for the sedative effects of intranasal dexmedetomidine was −2.45 (random, 95% CI; −3.33, −1.58) for continuous outcomes and RR of unsatisfactory patient outcome was 0.42 (M-H, random 95% CI; 0.26, 0.68 I 2 =45%) for dichotomous outcomes compared to that of intranasal saline. The SMD for the sedative effects of intranasal dexmedetomidine was −0.41 (random, 95% CI; −1.09, 0.27 I 2 =69%) for continuous outcomes and RR was 0.43 (M-H, random 95% CI; 0.32, 0.58 I 2 =0%) for dichotomous outcomes compared to that of per os benzodiazepines. Conclusions This review suggests that intranasal dexmedetomidine is associated with better sedative effects than oral benzodiazepines without producing respiratory depression, but it had a significantly delayed onset of effects.

Background Obesity develops when dietary energy intake exceeds energy expenditure, and can be associated with metabolic syndrome. Recent studies have shown that dietary phytochemicals can promote energy expenditure by inducing the browning of white adipose tissue (WAT). Purpose This study investigated whether cardamonin induces the browning of 3T3-L1 adipocytes through the activation of protein kinase A (PKA). Methods Anti-obesity potential of cardamonin was evaluated in 3T3-L1 adipocytes. Adipocyte-specific genes were observed using western blot, qPCR analysis and immunocytochemistry. Results Cardamonin treatment inhibited lipid droplet accumulation and reduced the expression of the adipogenic proteins C/EBPα and FABP4, and the lipogenic proteins LPAATθ, lipin 1, DGAT1, SREBP1, and FAS. Cardamonin also induced the expression of the browning marker genes PRDM16, PGC1α, and UCP1 at the mRNA and protein levels, and induced mRNA expression of CD137, a key marker of beige adipocytes. It also increased the expression of the β-oxidation genes CPT1 and PPARα at the mRNA and protein levels. In addition, cardamonin increased PKA phosphorylation and the mRNA and protein expression of the downstream lipolytic enzymes adipose triglyceride lipase (ATGL) and hormone‐sensitive lipase (HSL). Conclusion Our findings demonstrate novel effects of cardamonin to stimulate adipocyte browning, suppress lipogenesis, and promote lipolysis, implying it may have potential as an anti-obesity agent.

Drowning is one of the major causes of traumatic death. The impact of drowning in the elderly and patients who were not elderly will be different because of physiological differences. We wanted to analyze the clinical differences such as mortality, incidence rate of complications, degree of hypothermia and rate of cardiac arrest between elderly and adult drowning patients.This study included drowning patients over 18 years old who came to an emergency department (ED) located on a riverside from September 1997 to July 2016. Patients over the age of 65 years were classified as elderly, while those under the age of 65 years were classified as adults. Demographic data and clinical outcomes were surveyed.A total of 611 patients were included in this study. Sixty-one patients (9.9%) were elderly, and 550 patients (90.1%) were adults. There were 17 elderly patients (15.8%) and 87 adult patients (27.9%) who had cardiac arrest at the time of ED arrival (p = 0.017). The rate of body temperatures 34 °C was higher in elderly patients than that in adult patients (27.9% vs 17.5%, respectively, p = 0.025). The rates of hospitalization in the intensive care unit (ICU) and mortality were higher in elderly group (23% vs. 15.1%, respectively, p = 0.01; 37.7% vs 21.8%, respectively, p = 0.01). There was no significant difference in suicidal intent between the elderly and adult patient groups (82.0% vs 78.9%, respectively, p = 0.421).Elderly drowning patients accounted for approximately 1/10 of all drowning cases and were more likely to experience a cardiac arrest, hypothermia, mortality, and ICU admission.

Alzheimer’s disease (AD) is an age-related disorder that causes a loss of brain function. Hyperphosphorylation of tau and the subsequent formation of intracellular neurofibrillary tangles (NFTs) are implicated in the pathogenesis of AD. Hyperphosphorylated tau accumulates into insoluble paired helical filaments that aggregate into NFTs; therefore, regulation of tau phosphorylation represents an important treatment approach for AD. Heat shock protein 27 (Hsp27) plays a specific role in human neurodegenerative diseases; however, few studies have examined its therapeutic effect. In this study, we induced tau hyperphosphorylation using okadaic acid, which is a protein phosphatase inhibitor, and generated a fusion protein of Hsp27 and the protein transduction domain of the HIV Tat protein (Tat-Hsp27) to enhance the delivery of Hsp27. We treated Tat-Hsp27 to SH-SY5Y neuroblastoma cells for 2 h; the transduction level was proportional to the Tat-hsp27 concentration. Additionally, Tat-Hsp27 reduced the level of hyperphosphorylated tau and protected cells from apoptotic cell death caused by abnormal tau aggregates. These results reveal that Hsp27 represents a valuable protein therapeutic for AD. Electronic supplementary material The online version of this article (doi:10.1007/s10571-015-0199-1) contains supplementary material, which is available to authorized users.

Allergy testing is important for evidence-based care and treatment. In this paper, we report a new clinical laboratory instrument, AdvanSure AlloStation Smart, which is used for a multiple allergen-specific immunoglobulin (IgE) assay. For efficient automation of immunoblot procedures, a novel tilting carousel technology was incorporated. The analyzer showed 100.5–101.7% of pipetting accuracy, no sample carryover, and 0.2–3.2% coefficient of variation of optical precision. Assay performance on the system was comparable with that of a single allergen test, ImmunoCAP for 23 allergens. The slope and the intercept of this comparison were 0.994 and 0.070, respectively. The Pearson correlation coefficient between two methods was r = 0.836. The coefficient of variation of intra-assay precision ranged from 3.2% to 11.1% and that of inter-assay precision varied from 4.0% to 17.6%. The AdvanSure AlloStation Smart provides a simple and reliable multiple assay platform to monitor allergen-specific IgE profiles.

BACKGROUND Preoperative anxiety may differ according to patient temperament. It will be increased when patients are requested to participate in a study involving anesthesia. The purpose of this study was to show that the anxiety felt when patients are requested to participate may differ according to temperament in both patients who agree and disagree to participate. METHODS Three hundred and twenty-one patients over age 18 with American Society of Anesthesiologists 1 and 2 completed a survey questionnaire. The degree of anxiety was measured according to patient temperament. It was compared on the basis of the State-Trait Anxiety Inventory (STAI) and visual analogue scale (VAS). RESULTS In the agreed group, the degree of anxiety measured by "usual, present STAI" and VAS in the monitors (those who want to know as much as possible about anesthesia and surgery) was significantly higher than that in the blunters (those who want to know as little as possible) (P = 0.041 for the "usual STAI", 0.017 for "present STAI", and 0.001 for VAS, respectively). Among patients with a lower educational level, the numbers of blunters and monitors were 57 (79%) and 32 (59%), respectively, indicating that the ratio of blunters was significantly higher (P = 0.026). CONCLUSIONS Both traits of patients in each group were influenced by psychological burdens. The anxiety of the monitors who agreed to participate was significantly higher than that of blunters. In addition to temperament, education level affects participation. Obtaining consent for participation by understanding temperament and considering factors that may reduce the participation rate will be required.

The higher and bigger buildings are, the huger the scale of fire is. As the height of buildings gets greater, the amount of lightweight concrete increases to be used for dividers and separation wall taking the weight of the building into consideration. In particular, in the case of using only materials for weight lightening, at a fire, expansion, explosive fracture and emission of toxic gas would occur, so damage of life and property is concerned. For this reason, in this research, we studied light weight materials which used Bottom Ash, a byproduct of a thermoelectric power plant, as main materials, instead of imported light weight materials. Through the manufacturing technology coating more than 5 mm Bottom Ash particle with less than 3 mm Bottom Ash particle, the air hole, the advantage of Bottom Ash, is maintained, and the absorption, the disadvantage of it, is improved. As a result, it satisfied all the requirement such as the absorption, compressive strength, and unit weight like the basic light weight concrete panel, and the absorption is improved by more than 50%, compared to Non-Coating Bottom Ash. In the experiment of incombustibility, we also studied great manufacturing technology of light-weight-concrete passed through KS F ISO 1182,2271(incombustible materials).

Substation Automation System (SAS) based on IEC 61850 is becoming more and more reality in applying power systems. As well, the development of various engineering tools and the convergence with diagnostic systems are underway. To reflect recent advancements in SAS technology, the Korea Electric Power Corporation initiated a 154kV SAS pilot project and a smart power grid business in 2010. In this paper, the authors will summarize the overview and the lessons learned in applying the IEC 61850-based 154kV Main transformer bay IED solutions to these two project.

The rapid growth of e-business market makes new online companies to start and existing offline companies to join in this area. As the number of players of this market grows rapidly, the competition among them is very intense. Many companies invest huge resources to online marketing including search advertisement, email advertisement and banner advertisement. Because these traditional online marketing activities mainly focus on how to invite visitors to their web sites, ROI of these marketing activities are getting lower. Many companies are looking for a new marketing method to escape this situation. In this paper, we propose ROMS (Realtime Online Marketing System) which supports tools to improve conversion ratio of e-commerce sites, ROMS gathers behavioral data of visitors and analyzes it in realtime. ROMS supports live chats, visitor profiling, context analysis, event detection, and live marketing. With ROMS, personalized offers based on visitors` realtime context can be made for each visitor.

The purpose of this study was to determine the late decline in viability of fat cells over time for fat tissue stored at -15 degrees C and -70 degrees C after harvest from abdominal liposuction. A total of 16 females were recruited for this study. The viability of fat cell specimens was measured after freezing for 1, 3, 7, 14, 28, and 56 days. A number of viable mature adipocytes were evaluated by fluorescence microscopy after staining with fluorescein diacetate and propidium iodide. The glycerol-3-phosphate dehydrogenase activity was measured in lipoaspirates before digestion and the XTT reduction assay was performed. In addition, the XTT reduction assay was also performed on isolated lipocytes and preadipocytes.The viability of mature adipocytes was very low for both the -15 degrees C and -70 degrees C samples after 1 day of freezing (13.3% +/- 7.4% and 12.6% +/- 6.3%, respectively). There was no statistically significant difference between the samples stored at the 2 temperatures. The GPDH activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 25.1% +/- 10% and 28.7% +/- 11%, respectively. For the XTT test, the fractional enzyme activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 30.0% +/- 10.9% and 36.1% +/- 12.3%, respectively. In addition, the adipocytes had low activity from day one: 15.4% +/- 7.2% at -15 degrees C and 11.5% +/- 5.6% at -70 degrees C. Furthermore, the preadipocytes had a low activity of 8.0% +/- 6.0% at -15 degrees C and 8.6% +/- 3.8% at -70 degrees C. At 8 weeks, there were few viable mature adipocytes and the activity of the cells was very low by XTT and GPDH testing.The results of this study showed that the viability of adipocytes declined rapidly after frozen storage for 1 day at both -15 degrees C and -70 degrees C, and decreased gradually in storage after 8 weeks; at which time only approximately 5% of the fat cells were alive. These findings suggest that the present fat preservation storage techniques using a -15 degrees C freezer or a -70 degrees C deep freezer are both inadequate to maintain the viability of fat cells.

An amphipathic leucine (L) and lysine (K)‐rich α‐helical peptide is multimerized based on helix‐loop‐helix structures to maximize the penetrating activities. The multimeric LK‐based cell penetrating peptides (LK‐CPPs) can penetrate cells as protein‐fused forms at 100–1000‐fold lower concentrations than Tat peptide. The enhanced penetrating activity is increased through multimerization by degrees up to the tetramer level. The multimeric LK‐CPPs show rapid cell penetration through macropinocytosis at low nanomolar concentrations, unlike the monomeric LK, which have slower penetrating kinetics at much higher concentrations. The heparan sulfate proteoglycan (HSPG) receptors are highly involved in the rapid internalization of multimeric LK‐CPPs. As a proof of concept of biomedical applications, an adipogenic transcription factor, peroxisome proliferator‐activated receptor gamma 2 (PPAR‐γ 2), is delivered into preadipocytes, and highly enhanced expression of adipogenic genes at nanomolar concentrations is induced. The multimeric CPPs can be a useful platform for the intracellular delivery of bio‐macromolecular reagents that have difficulty with penetration in order to control biological reactions in cells at feasible concentrations for biomedical purposes.

The antibody levels against the C-terminal region of the merozoite surface protein 1 of Plasmodium vivax (PvMSP1c) were measured in 276 patients with P. vivax malaria (patient group), 320 malaria-naïve healthy individuals (control group 1), and 70 malaria-naïve individuals with various disorders (control group 2) using the immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) and the direct sandwich ELISA. To evaluate the antibody response during relapse, 5 relapsed patients were tested using the IgM capture ELISA. The IgM antibodies were negative in 99.7% of control group 1 and in 100% of control group 2; they were positive in 90.6% of the patient group. The total antibody levels were positive in 88.4% of the patient group with the direct sandwich ELISA. The sera from the second malaria episode, i.e., relapsed patients, were 100% positive on the IgM capture ELISA. The results of this study suggest that the IgM capture ELISA may be a useful diagnostic method for P. vivax malaria for both primary infection and relapse.

Although diagnosis of Plasmodium vivax malaria has been difficult when it is present at a low parasite density, it was recently revealed that an antibody assay was a good method of screening for malaria in blood banks. However, the use of this method for the diagnosis of malaria is limited due to the persistence of specific immunoglobulin (Ig) G. Therefore, we evaluated specific IgM antibody responses against the C-terminal region of the merozoite surface protein 1 of P. vivax (PvMSP1c) in sera obtained from patients with vivax malaria using various assays. The IgM capture enzyme-linked immunosorbent assay showed good sensitivity (97.7%; 308/315) and specificity (99.1%, 446/450). In addition, the results of this assay were not related to parasite density, and a high reactivity was observed when there was a low level of parasitemia. Furthermore, we found that patients with cases of malaria that had relapsed still had the IgM titers against PvMSP1c. Therefore, the use of IgM ELISA for the detection of specific IgM that was not involved in memorial immune activity could be an alternative tool for the diagnosis of malaria and blood screening, even in areas in which malaria is endemic.

We expressed a C-terminal 108-aa region of the Plasmodium vivax merozoite surface protein 1 (PvMSP1c) excluding the C-end transmembrane region in Escherichia coli in order to evaluate the antibody level to MSP in Korean malaria patients. We optimized a direct sandwich enzyme-linked immunosorbent assay (ELISA) method to simultaneously determine the total antibody levels, including IgG and IgM, to PvMSP1c. If the cut-off for seropositivity was determined as the mean+3SD of the antibody levels of the negative control group, the antibody levels were positive in 99.5% of the patient group (sensitivity 199/200). The antibody levels were negative in 99.4% of the negative control group (specificity 504/507). The positive reactions in the negative control group came from non-specific reactions, as confirmed by a competition assay. This direct sandwich ELISA for PvMSP1c antibody could prove to be a useful tool for the diagnosis of malaria patients and for blood screening in blood banks.

The existing approaches to the detection of precipitation are largely based on room temperature measurements on quenched samples. However, the direct measurement of precipitation kinetics at high temperatures is also possible through analysis of the mechanical properties of the specimens. In this work, a creep method was developed and applied to the detection of aluminum nitride precipitation in a dual phase 3 % silicon steel containing 0.038 % C. Prior to loading, the specimens were solution treated for 20 min and then cooled to the test temperature. A constant stress was applied to each sample by means of a computerized MTS machine and the strain was recorded continuously during testing. Microstructural examination revealed that the austenite fraction and morphology and the microstructure of the ferrite matrix are quite different depending on whether samples are directly heated or heated and then cooled to the test temperature. The resulting creep rate is sensitive both to the occurrence of precipitation as well as to phase transformation; when the microstructure remains fixed, the slope of the true strain-log(time) curve decreases immediately after the initiation of precipitation. The precipitation-time-temperature diagrams determined in this way are of classical C shape.

This paper presents the development of Bay Control Unit for Power Transformer to build up substation automation system. The substation automation including digital control and monitoring of electric machine is widely spreading more and more, the degree of automation is more sophisticated and automation functions are required increasingly. These requirements of related power industries and customers can be satisfied by advanced digital IEDs such as digital protection relay, bay controller, PLC, communication devices, etc. By the fusion of state of the art technologies such as microprocessor, digital control, digital communication with conventional power industry, we can build a more effective, reliable, economic, convenient substation and power system.