Bone Reconstruction Using Two-Layer Porcine-Derived Bone Scaffold Composed of Cortical and Cancellous Bones in a Rabbit Calvarial Defect Model.

In this study, we aimed to investigate the bone regeneration efficiency of two-layer porcine-derived bone scaffolds composed of cancellous and cortical bones in a rabbit calvarial defect model. Four circular calvaria defects were formed on cranium of rabbit and were filled with block bone scaffolds of each group: cortical bone block (Cortical group), cancellous bone block (Cancellous group), and two-layer bone block (2layer group). After 8 weeks, new bones were primarily observed in cancellous parts of the Cancellous and 2layer groups, while the Cortical group exhibited few new bones. In the results of new bone volume and area analyses, the Cancellous group showed the highest value, followed by the 2layer group, and were significantly higher than the Cortical group. Within the limitations of this study, the cancellous and two-layer porcine-derived bone scaffolds showed satisfactory bone regeneration efficiency; further studies on regulating the ratio of cortical and cancellous bones in two-layer bones are needed.

Elevated Expression of Cathepsin K in Periodontal Ligament Fibroblast by Inflammatory Cytokines Accelerates Osteoclastogenesis via Paracrine Mechanism in Periodontal Disease.

Cathepsin K (CTSK) is a cysteine protease that is mainly produced from mature osteoclasts and contributes to the destruction of connective tissues and mineralized matrix as a consequence of periodontal disease (PD). However, few studies have reported its regulatory role in osteoclastogenesis-supporting cells in inflammatory conditions. Here, we investigated the role of CTSK in osteoclastogenesis-supporting cells, focusing on the modulation of paracrine function. Microarray data showed that CTSK was upregulated in PD patients compared with healthy individuals, which was further supported by immunohistochemistry and qPCR analyses performed with human gingival tissues. The expression of CTSK in the osteoclastogenesis-supporting cells, including dental pulp stem cells, gingival fibroblasts, and periodontal ligament fibroblasts (PDLFs) was significantly elevated by treatment with inflammatory cytokines such as TNFα and IL-1ß. Moreover, TNFα stimulation potentiated the PDLF-mediated osteoclastogenesis of bone marrow-derived macrophages. Interestingly, small interfering RNA-mediated silencing of CTSK in PDLF noticeably attenuated the TNFα-triggered upregulation of receptor activator of nuclear factor kappa-B ligand (RANKL), macrophage colony-stimulating factor, and RANKL/osteoprotegerin ratio, thereby abrogating the enhanced osteoclastogenesis-supporting activity of PDLF. Collectively, these results suggest a novel role of CTSK in the paracrine function of osteoclastogenesis-supporting cells in periodontal disease.

Clinical Implications of Ventricular Repolarization Parameters on Long-Term Risk of Atrial Fibrillation　- Longitudinal Follow-up Data From a General Ambulatory Korean Population.

BACKGROUND: This study investigated 12-lead electrocardiogram (ECG) predictors associated with atrial fibrillation (AF) or flutter (AFL), specifically whether ventricular repolarization abnormalities in surface ECG (i.e., non-specific ST-T abnormalities [NSSTTA], QT prolongation, early repolarization [ER]) were associated with the development of AF or AFL.Methods and Results:This study included 16,793 ambulatory Koreans (mean age 48.2 years, 62.3% male) who underwent medical check-ups at Asan Medical Center in 2002 (NSSTTA, n=1,037 [6.2%]; ER, n=1,493 [8.9%]). The primary outcome was the incidence of ECG-documented AF or AFL. During follow-up, new-onset AF or AFL was documented in 334 subjects (2.0%). The incidence of AF or AFL at the 10-year follow-up was higher in patients with than without NSSTTA (3.5% vs. 1.6%; hazard ratio [HR] 1.79, 95% confidence interval [CI] 1.28-2.50). The QT interval was associated with a higher risk of AF or AFL (HR 1.12 [95% CI 1.07-1.17] per 10 ms), and the risk was even higher in patients with multiple-region NSSTTA (HR 2.30; 95% CI 1.64-3.21) and NSSTTA with QT prolongation (HR 4.06; 95% CI 2.14-7.69). ER was not associated with a higher risk of AF or AFL (HR 1.02; 95% CI 0.71-1.46). CONCLUSIONS: NSSTTA and QT prolongation, but not ER, were associated with a higher risk of future AF or AFL in a general ambulatory population after adjusting for parameters of atrial depolarization.

Multivalent-Interaction-Driven Assembly of Discrete, Flexible, and Asymmetric Supramolecular Protein Nano-Prisms.

Current approaches to design monodisperse protein assemblies require rigid, tight, and symmetric interactions between oligomeric protein units. Herein, we introduce a new multivalent-interaction-driven assembly strategy that allows flexible, spaced, and asymmetric assembly between protein oligomers. We discovered that two polygonal protein oligomers (ranging from triangle to hexagon) dominantly form a discrete and stable two-layered protein prism nanostructure via multivalent interactions between fused binding pairs. We demonstrated that protein nano-prisms with long flexible peptide linkers (over 80 amino acids) between protein oligomer layers could be discretely formed. Oligomers with different structures could also be monodispersely assembled into two-layered but asymmetric protein nano-prisms. Furthermore, producing higher-order architectures with multiple oligomer layers, for example, 3-layered nano-prisms or nanotubes, was also feasible.

Cutting Edge: LL-37-Mediated Formyl Peptide Receptor-2 Signaling in Follicular Dendritic Cells Contributes to B Cell Activation in Peyer's Patch Germinal Centers.

Peyer's patches (PPs) are the major mucosal immune-inductive site, and germinal centers (GCs) in PPs determine the quality of the Abs produced. PP GCs are continuously induced by the gut microbiota, and their maintenance contributes to the induction of strong IgA responses to Ags. In this study, we investigated the role of formyl peptide receptor (FPR)-mediated signaling in the maintenance of PP GCs, because FPRs recognize the microbiota and initiate an innate immune response by chemotaxis. We found that follicular dendritic cells (FDCs), a key organizer of B cell follicles and GCs in mucosal immunity, express Fpr2. Additionally, Fpr2-mediated signaling in PP FDCs promoted Cxcl13 and B cell activating factor expression, as well as B cell proliferation and activation. Therefore, we suggest that Fpr2-mediated signaling in FDCs plays a key role in GC maintenance in PPs and results in an Ag-specific IgA response in the gut mucosal immune compartment.

C5a receptor targeting of partial non-structural protein 3 of dengue virus promotes antigen-specific IFN-γ-producing T-cell responses in a mucosal dengue vaccine model.

Mucosal vaccination is an ideal strategy to induce protective immunity in both mucosal and parenteral areas. Successful induction of an antigen-specific immune response via mucosal administration essentially requires the effective delivery of antigen into a mucosal immune inductive site, which depends on antigen delivery into M cells. We previously reported that M cells specifically express C5aR, and antigen targeting to C5aR by using specific ligands, including Co1 peptide, promotes the antigen-specific immune response in both mucosal and systemic immune compartments. In this study, we found that application of the Co1 peptide to dengue virus antigen containing CD8 T cell epitopes effectively induced an antigen-specific IFN-γ-producing CD8+ T cell response after oral mucosal administration of antigen. Consequently, we suggest that Co1 peptide-mediated C5aR targeting of antigen into M cells can be used for the induction of an effective antigen-specific CD8+ T cell immune response in oral mucosal vaccine development.

Counterion-Induced Control of the Colloidal State of Polyamic Acid Nanoparticles for Electrophoretic Deposition.

Optimizing the colloidal state of polyamic acid (PAA) nanoparticles is essential for achieving a uniform and high-performance polyimide coating by electrophoretic deposition (EPD) on metal substrates of various shapes. In this paper, we report two important roles of the counterions in the formation of PAA colloids for EPD, which, to date, have not been recognized. First, when tertiary alkyl amines are used to neutralize PAA, the polarity of neutralizing counterions determines the size and stability of the PAA colloidal particles. The polarity can be finely tuned by using two different tertiary alkyl amines containing polar and nonpolar groups and adjusting the molar ratio. Depending on the polar/nonpolar ratio, various states of PAA colloids were obtained, including dissolved state, stable colloid, and aggregates. Second, we observed that the confined counterions inside PAA nanoparticles can act as an imidization catalyst during the thermal annealing process. It is revealed that some fraction of the counterion species, mostly having nonpolar groups, is not drawn toward the counter electrode and remains inside the PAA nanoparticles during the EPD process. Optimizing the polarity eventually allowed us to form uniform EPD coatings with high dielectric strengths.

Development of a monoclonal antibody specific to envelope domain III with broad-spectrum detection of all four dengue virus serotypes.

Dengue virus (DENV) is a mosquito-borne pathogen that annually infects more than 390 million people in 100 different countries. Symptoms of the viral infection include a relatively weak dengue fever to severe dengue hemorrhagic fever/dengue shock syndrome, which are mortal infectious diseases. As of yet, there is no commercially available vaccine or therapeutic for DENV. Currently, passive immunotherapy using DENV-specific antibody (Ab) is a considered strategy to treat DENV infection. Here, we developed a monoclonal Ab (mAb), EDIIImAb-61, specific to the DENV domain III of the envelope glycoprotein (EDIII) with broad-spectrum detection ability to all four DENV serotypes (DENV-1∼4) to use as a therapeutic Ab. Although EDIII contains non-immunodominant epitopes compared to domains I and II, domain III plays a critical role in host receptor binding. EDIIImAb-61 exhibited cross-reactive binding affinity to all four DENV serotypes that had been isolated from infected humans. To further characterize EDIIImAb-61 and prepare genes for large-scale production using a heterologous expression system, the sequence of the complementarity determining regions was analyzed after cloning the full-length cDNA genes encoding the heavy and light chain of the mAb. Finally, we produced Ab from CHO-K1 cells transfected with the cloned EDIIImAb-61 heavy and light chain genes and confirmed the binding ability of the Ab. Collectively, we conclude that EDIIImAb-61 itself and the recombinant Ab produced using the cloned heavy and light chain gene of EDIIImAb-61 is a candidate for passive immunotherapy against DENV infection.

Artificial supramolecular protein assemblies as functional high-order protein scaffolds.

Supramolecular assemblies of protein building blocks potentially offer unique biomaterials with unmatched functionalities as well as atomic level structural accuracy. An increasing number of assembling strategies have been reported for the fabrication of diverse artificial protein assemblies, ranging from rather heterogeneous protein oligomers to computationally designed discrete protein architectures. In this perspective, we discuss these artificial protein supramolecules in terms of their use as highly potent high-order protein scaffolds that can display various functional proteins with precise structural and valency control. Following a brief overview of current approaches for protein assembly, several examples of functional protein assemblies have been introduced, with a particular focus on our recent report of valency-controlled green fluorescent protein nano-assemblies. Our supramolecular protein scaffolds allow building a series of polygonal assemblies of functional binding proteins, which provide unprecedented ways to study multivalent protein interactions. Even with many remaining challenges, there is unlimited potential of artificial protein scaffolds in many fields from nanotechnology to vaccine development.

Switching Mechanisms of Nonvolatile Memory Devices Fabricated with a Polydopamine Layer.

Nonvolatile memory devices based on a polydopamine (PDA) layer were fabricated by using a dip-coating process. Atomic force microscopy images revealed that the PDA layer had a conformal surface. The energy dispersive X-ray data showed the atomic stoichiometry of nitrogen and carbon in the PDA layer. The capacitance-voltage (C-V) curves of the Al/PDA/n-Si memory devices at 300 K showed a hysteresis with a large flat band shift, indicating that the incomplete PDA layer acted as a charge storage in the memory device. The switching mechanisms for the writing and erasing processes for the Al/PDA/n-Si devices are described on the basis of the C-V results and the energy band diagrams.

The hsSsu72 phosphatase is a cohesin-binding protein that regulates the resolution of sister chromatid arm cohesion.

Cohesin is a multiprotein complex that establishes sister chromatid cohesion from S phase until mitosis or meiosis. In vertebrates, sister chromatid cohesion is dissolved in a stepwise manner: most cohesins are removed from the chromosome arms via a process that requires polo-like kinase 1 (Plk1), aurora B and Wapl, whereas a minor amount of cohesin, found preferentially at the centromere, is cleaved by separase following its activation by the anaphase-promoting complex/cyclosome. Here, we report that our budding yeast two-hybrid assay identified hsSsu72 phosphatase as a Rad21-binding protein. Additional experiments revealed that Ssu72 directly interacts with Rad21 and SA2 in vitro and in vivo, and associates with sister chromatids in human cells. Interestingly, depletion or mutational inactivation of Ssu72 phosphatase activity caused the premature resolution of sister chromatid arm cohesion, whereas the overexpression of Ssu72 yielded high resistance to this resolution. Interestingly, it appears that Ssu72 regulates the cohesion of chromosome arms but not centromeres, and acts by counteracting the phosphorylation of SA2. Thus, our study provides important new evidence, suggesting that Ssu72 is a novel cohesin-binding protein capable of regulating cohesion between sister chromatid arms.

High-Resolution Nanotransfer Printing of Porous Crossbar Array Using Patterned Metal Molds by Extreme-Pressure Imprint Lithography.

High-resolution nanotransfer printing (nTP) technologies have attracted a tremendous amount of attention due to their excellent patternability, high productivity, and cost-effectiveness. However, there is still a need to develop low-cost mold manufacturing methods, because most nTP techniques generally require the use of patterned molds fabricated by high-cost lithography technology. Here, we introduce a novel nTP strategy that uses imprinted metal molds to serve as an alternative to a Si stamp in the transfer printing process. We present a method by which to fabricate rigid surface-patterned metallic molds (Zn, Al, and Ni) based on the process of direct extreme-pressure imprint lithography (EPIL). We also demonstrate the nanoscale pattern formation of functional materials, in this case Au, TiO2, and GST, onto diverse surfaces of SiO2/Si, polished metal, and slippery glass by the versatile nTP method using the imprinted metallic molds with nanopatterns. Furthermore, we show the patterning results of nanoporous crossbar arrays on colorless polyimide (CPI) by a repeated nTP process. We expect that this combined nanopatterning method of EPIL and nTP processes will be extendable to the fabrication of various nanodevices with complex circuits based on micro/nanostructures.

Vasohibin-1 promotes osteoclast differentiation in periodontal disease by stimulating the expression of RANKL in gingival fibroblasts.

Vasohibin-1 (VASH1) is a key inhibitor of vascular endothelial growth factor-induced angiogenesis. Although the involvement of VASH1 in various pathological processes has been extensively studied, its role in periodontal disease (PD) remains unclear. We aimed to investigate the role of VASH1 in PD by focusing on osteoclastogenesis regulation. We investigated VASH1 expression in PD by analyzing data from the online Gene Expression Omnibus (GEO) database and using a mouse ligature-induced periodontitis model. The effects of VASH1 on osteoclast differentiation and osteoclastogenesis-supporting cells were assessed in mouse bone marrow-derived macrophages (BMMs) and human gingival fibroblasts (GFs). To identify the stimulant of VASH1, we used culture broth from Porphyromonas gingivalis (Pg), a periopathogen. The GEO database and mouse periodontitis model revealed that VASH1 expression was upregulated in periodontitis-affected gingival tissues, which was further supported by immunohistochemistry and qRT-PCR analyses. VASH1 expression was significantly stimulated in GFs after treatment with the Pg broth. Direct treatment with recombinant VASH1 protein did not stimulate osteoclast differentiation in BMMs but did contribute to osteoclast differentiation by inducing RANKL expression in GFs through a paracrine mechanism. Small interfering RNA-mediated silencing of VASH1 in GFs abrogated RANKL-mediated osteoclast differentiation in BMMs. Additionally, VASH1-activated RANKL expression in GFs was significantly suppressed by MK-2206, a selective inhibitor of AKT. These results suggest that Pg-induced VASH1 may be associated with RANKL expression in GFs in a paracrine manner, contributing to osteoclastogenesis via an AKT-dependent mechanism during PD progression.

Formation of Multiscale Pattern Structures by Combined Patterning of Nanotransfer Printing and Laser Micromachining.

Various lithography techniques have been widely used for the fabrication of next-generation device applications. Micro/nanoscale pattern structures formed by lithographic methods significantly improve the performance capabilities of the devices. Here, we introduce a novel method that combines the patterning of nanotransfer printing (nTP) and laser micromachining to fabricate multiscale pattern structures on a wide range of scales. Prior to the formation of various nano-in-micro-in-millimeter (NMM) patterns, the nTP process is employed to obtain periodic nanoscale patterns on the target substrates. Then, an optimum laser-based patterning that effectively engraves various nanopatterned surfaces, in this case, spin-cast soft polymer film, rigid polymer film, a stainless still plate, and a Si substrate, is established. We demonstrate the formation of well-defined square and dot-shaped multiscale NMM-patterned structures by the combined patterning method of nTP and laser processes. Furthermore, we present the generation of unusual text-shaped NMM pattern structures on colorless polyimide (CPI) film, showing optically excellent rainbow luminescence based on the configuration of multiscale patterns from nanoscale to milliscale. We expect that this combined patterning strategy will be extendable to other nano-to-micro fabrication processes for application to various nano/microdevices with complex multiscale pattern geometries.

The Role of Physiotherapy in the Management of Functional Neurological Disorder in Children and Adolescents.

Children and adolescents with functional neurological (conversion) disorder (FND) present with symptoms of impaired motor and sensory function. FND involves complex interactions between the brain, mind, body, and lived experience of the child. The gold standard for treatment is therefore a holistic, biopsychosocial approach with multimodal interventions delivered by a multidisciplinary team. In this narrative review we examine the role of physiotherapy in managing FND in children. We searched Embase, Medline, PsycINFO, and PubMed (back to 2000) for relevant physiotherapy articles and also manually searched their reference lists. Two review articles and ten observational studies were identified. Data were extracted concerning the type of study, therapies involved, outcome measures, and comorbid mental health outcomes. FND symptoms resolved in 85% to 95% of the patients, and about two-thirds returned to full-time school after completing the multidisciplinary intervention. Ongoing mental health concerns at follow-up were associated with poorer functional outcomes. Key themes included the following: use of psychological interventions embedded in the physiotherapy intervention; integration of play, music, and dance; role of physical exercise in modulating physiological, neural, and endocrine systems; need for FND-specific outcome measures; ethical issues pertaining to randomized trials; and need to develop alternate study methodologies for assessing combined treatments. Clinical vignettes were included to highlight a range of physiotherapy interventions. In conclusion, the emerging literature suggests that physiotherapy for children with FND is a useful intervention for improving motor dysfunction and for addressing other concurrent issues such as physical deconditioning, neuroprotection, chronic pain, disturbed sleep, anxiety and depression, and resilience building.

Virulence factors released from Porphyromonas gingivalis induce electrophysiological dysfunction in human pluripotent stem cell-derived cardiomyocytes.

Background/purpose: Periodontal disease development correlates with the occurrence of systemic diseases. The present study investigated the association between periodontal disease and the development of cardiac arrhythmia. Materials and methods: Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) were treated with Porphyromonas gingivalis (Pg). Cardiotoxicity and electrophysiological properties of hESC-CMs were measured using the cell counting kit-8 assay and a multi-electrode array, respectively. Reverse-transcription-quantitative polymerase chain reaction (RT-qPCR) revealed the mRNA expression of S100 calcium binding protein A1 (S100A1), calsequestrin 2 (CASQ2), troponin I3 (TNNI3), myosin light chain 2 (MYL2), integrin subunit beta 1 (ITGB1), and cadherin 2 (CDH2) in hESC-CMs. Results: Treatment with Pg broth significantly decreased the beat period, field potential duration, spike amplitude, and conduction velocity without affecting the viability of hESC-CMs. In addition, the mRNA expression of CASQ2, TNNI3, and MYL2, which are all associated with calcium handling, were downregulated by Pg broth treatment. Conclusion: These findings indicate that Pg may induce cardiac arrhythmia mediated by virulence factors.

Immunomodulatory Effect of Hispolon on LPS-Induced RAW264.7 Cells and Mitogen/Alloantigen-Stimulated Spleen Lymphocytes of Mice.

Hispolon is a potent anticancer, anti-inflammatory, antioxidant, and antidiabetic agent isolated from Phellinus linteus, an oriental medicinal mushroom. However, the immunomodulatory mechanisms by which hispolon affects macrophages and lymphocytes remain poorly characterized. We investigated the immunomodulatory effects of hispolon on oxidative stress, inflammatory responses, and lymphocyte proliferation using lipopolysaccharide (LPS)-treated RAW264.7 macrophages or mitogen/alloantigen-treated mouse splenocytes. Hispolon inhibited LPS-induced reactive oxygen and nitrogen species (ROS/RNS) generation and decreased total sulfhydryl (SH) levels in a cell-free system and RAW264.7 cells. Hispolon exerted significant anti-inflammatory effects by inhibiting production of the proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) and activation of nuclear factor kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3) in LPS-treated RAW264.7 cells. Hispolon also modulated NF-κB and STAT3 activation by suppressing the NF-κB p65 interaction with phospho-IκBα and the STAT3 interaction with JAK1, as determined via coimmunoprecipitation analysis. Additionally, hispolon significantly decreased lymphocyte proliferation, T cell responses and T helper type 1 (Th1)/type 2 (Th2) cytokines production in mitogen/alloantigen-treated splenocytes. We conclude that hispolon exerts immunomodulatory effects on LPS-treated macrophages or mitogen/alloantigen-treated splenocytes through antioxidant, anti-inflammatory, and antiproliferative activities. Thus, hispolon may be a therapeutic agent for treating immune-mediated inflammatory diseases.

Oriented Crystal Growth during Perovskite Surface Reconstruction.

Surface passivation has become a key strategy for an improvement in power conversion efficiency (PCE) of perovskite solar cells (PSCs) since PSCs experienced a steep increase in PCE and reached a comparably matured point. Recently, surface passivation using a mixed salt of fluorinated alkyl ammonium iodide and formamidinium bromide demonstrated a remarkable improvement in both performance and stability, which can be tuned by the length of the alkyl chain. Nevertheless, the role of the alkyl chain in manipulating surface-limited crystal growth was not fully understood, preventing a further progress in interface control. In this study, we found that the length of the fluorine-substituted alkyl chain governed the crystal formation dynamics by manipulating surface tensions of different crystal orientations. The overall enhancement of the (001) plane, being the most favored, commonly resulted from the surface reformation of the perovskite film regardless of the chain length, while the highly oriented (001) over (111) was monitored with a particular chain length. The enhanced crystal orientation during surface recrystallization was responsible for the low trap density and thus effectively suppressed charge recombination at the interface, resulting in a considerable increase in open-circuit voltage and fill factor.

Trends and Determinants in Breastfeeding among Korean Women: A Nationwide Population-Based Study.

Many efforts have been launched to promote breastfeeding in Korea. However, breastfeeding trends and associated factors with breastfeeding in Korea remain unknown. This study aimed to examine trends and determinants in breastfeeding using the Korea National Health and Nutrition Examination Survey (KNHANES) (2010-2018). We analyzed data from the KNHANES V (2010-2012), VI (2013-2015), and VII (2016-2018). A total of 9232 women aged 19-49 years were included in this study. We performed multivariable logistic regression analyses to investigate breastfeeding prevalence trends and associated factors with breastfeeding. Compared to 2010-2012, the odds ratio associated with breastfeeding during 2013-2015 and 2016-2018 increased to 1.30 (95% confidence interval (CI): 1.11-1.51) and 1.40 (95% CI: 1.21-1.63), respectively. The breastfeeding rate was associated with 40-49 years (OR, 95% CI: 0.47, 0.34-0.64 compared to 19-29 years), richer and poorer income (1.20, 1.03-1.39 in richer group and 1.24, 1.05-1.46 in poorer group compared to richest group), education level (0.74, 0.65-0.86 in ≤12 years of education compared to ≥13 years of education), smoking status (1.77, 1.38-2.28 in non-smoking compared to smoking), and self-rated health (1.40, 1.14-1.70 in good and 1.20, 1.00-1.44 in average compared to bad). Education programs and policies such as the Baby-Friendly Hospital Initiative (BFHI) and mother-friendly workplaces are necessary to increase the rates of breastfeeding in these groups.

Cut-off value of serum homocysteine in relation to increase of coronary artery calcification.

A recent study reported that coronary artery calcification (CAC) and serum homocysteine were well associated; however, no report is available for the cut-off value of serum homocysteine according to increase of coronary-artery calcification volume score (CVS). The data of 469 out of 777 subjects in 1 health promotion center located in Seoul were selected after exclusion of the missing data of serum homocysteine and CVS. CVS was categorized into 2 groups: CVS=0 and CVS>0. Serum homocysteine according to the CVS groups was compared, and the cut-off value of serum homocysteine according to the increase of CVS (>0) was calculated using the receiver operating characteristic curve. Mean age was 54.5 years and the proportion of females was 22.2%. Mean serum homocysteine concentration and CVS were 11.2 µmol/L and 50.4, respectively. After adjustments for age and sex, serum homocysteine was associated with CVS (r=0.167, p=0.001), and Log(Homocysteine) also showed a significant difference according to the CVS groups. The cut-off value of serum homocysteine according to the increase of CVS (>0) was 9.45 µmol/L (area under the curve=0.569 (95% CI 0.512 to 0.625), p=0.015). The cut-off value of serum homocysteine was 9.45 µmol/L according to the increase of coronary-artery CVS.