Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells

Haeeun Shin(Haeeun Shin),Joon Ki Min(Joon Ki Min),Na Rae Kim(Na Rae Kim),Kyoung Yul Seo(Kyoung Yul Seo),Hee Seung Chin(Hee Seung Chin),Soyoung Lee(Soyoung Lee),Ji Won Jung(Ji Won Jung) 대한안과학회 2023 Korean Journal of Ophthalmology Vol.37 No.1

Purpose: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs. Methods: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated into HCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs were assessed in the presence of different concentrations of Y-27632. Results: Y-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs showed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. Conclusions: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs.

[Colorectal] The Survival Analyses for Colorectal Cancer Patients with Mitomycin-C Induced Severe Neutropenia after Hyperthermic Intraperitoneal Chemotherapy with Cytoreductive Surgery

Soyoung Shin,Jeonghyun Kang,Seung Hyuk Baik,Eun Jung Park 대한외과학회 2022 대한외과학회 학술대회 초록집 Vol.2022 No.11

Design of potent 9-mer antimicrobial peptide analogs of protaetiamycine and investigation of mechanism of antimicrobial action

Shin, Soyoung,Kim, Jin-Kyoung,Lee, Jee-Young,Jung, Ki-Woong,Hwang, Jae-Sam,Lee, Juneyoung,Lee, Dong Gun,Kim, Iksoo,Shin, Song Yub,Kim, Yangmee John Wiley Sons, Ltd. 2009 Journal of Peptide Science Vol.15 No.9

<P>Protaetiamycine is an insect defensin, a naturally occurring 43-amino-acid-residue antimicrobial peptide derived from the larvae of the beetle Protaetia brevitarsis. In a previous work that aimed at developing short antibiotic peptides, we designed 9-mer peptide analogs of protaetiamycine. Among them, RLWLAIGRG-NH<SUB>2</SUB> showed good antifungal activity against Candida albicans. In this study, we designed four 9-mer peptide analogs based on the sequence of RLWLAIGRG-NH<SUB>2</SUB>, in which Gly or Ile was substituted with Arg, Lys, or Trp to optimize the balance between the hydrophobicity and cationicity of the peptides and to increase bacterial cell selectivity. We measured their toxicity to bacteria and mammalian cells as well as their ability to permeabilize model phospholipid membranes. Substitution of Arg for Gly9 at the C-terminus (9Pbw1) resulted in two- to fourfold improvement in antibacterial activity. Further substitution of Gly7 with Lys (9Pbw2 and 9Pbw4) caused four- to eightfold improvement in the antibacterial activity without increase in cytotoxocity, while substitution of Gly7 with Trp (9Pbw3) increased cytotoxicity as well as antibacterial activity. The peptides 9Pbw2 and 9Pbw4 with the highest bacterial cell selectivity were not effective in depolarizing the membrane of Staphylococcus aureus cytoplasmic membranes and showed almost no leakage of a fluorescent dye entrapped within the vesicles. Gel-retardation experiments indicated that 9Pbw2 and 9Pbw4 inhibited the migration of DNA at concentrations >20 µM. Three positively charged residues at the C-terminus in 9Pbw2 and 9Pbw4 may facilitate effective penetration into the negatively charged phospholipid membrane of bacteria. The results obtained in this study suggest that the bactericidal action of our potent antibacterial peptides, namely 9Pbw2 and 9Pbw4, may be attributed to the inhibition of the functions of intracellular components after penetration of the bacterial cell membrane. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd.</P>

Development and Stability Evaluation of In- House Prepared External Quality Controls for Autoimmune Disease Tests

Soyoung Shin,박준홍 대한임상검사정도관리협회 2021 Journal of Laboratory Medicine And Quality Assuran Vol.43 No.2

Background: The control samples for external quality assessment (EQA) must be highly stable during the EQA process. This study aimed to evaluate the stability of in-house prepared external quality controls for autoimmune disease tests, including anti-nuclear antibody (ANA), anti-double stranded DNA immunoglobulin G antibody (anti-dsDNA), anti-thyroglobulin antibody (anti-Tg), and anti-thyroperoxidase antibody (anti-TPO). Methods: The in-house prepared external quality controls with two different levels (negative and positive for ANA and anti-dsDNA; low and high for anti- Tg; and anti-TPO) for each autoimmune disease test were prepared and stored at test temperatures of –20℃, 4℃, and room temperature (RT). The levels of each autoimmune disease test were measured at baseline and at 1 week, 2 weeks, 1 month, 3 months, and 6 months. The internal quality controls from the same batch run were also tested during the evaluation period. Results: The negative and positive controls for ANA were stable for 6 months, regardless of storage temperature. The positive control for anti-dsDNA tended to increase in level at 6 months (P =0.033). The low and high anti-Tg controls tended to decrease in levels at 3 months (P =0.033 and P =0.042, respectively). The low control of anti-TPO decreased in level at RT compared to –20℃ at 1 week (P =0.026) and the high anti-TPO controls tended to decrease in level at 3 months regardless of storage temperature such as –20℃, 4℃, and RT (P =0.041). Conclusions: The stability of in-house prepared external quality controls was influenced by different storage temperatures and durations. These results indicate that all EQA samples for autoimmune disease tests should be stored at –20℃ or 4℃ and be tested as soon as possible during the EQA process.

Targeted next-generation sequencing identifies a novel nonsense mutation in <i>SPTB</i> for hereditary spherocytosis : A case report of a Korean family

Shin, Soyoung,Jang, Woori,Kim, Myungshin,Kim, Yonggoo,Park, Suk Young,Park, Joonhong,Yang, Young Jun Williams & Wilkins Co 2018 Medicine Vol.97 No.3

<P><B>Abstract</B></P><P><B>Rationale:</B></P><P>Hereditary spherocytosis (HS) is an inherited disorder characterized by the presence of spherical-shaped red blood cells (RBCs) on the peripheral blood (PB) smear. To date, a number of mutations in 5 genes have been identified and the mutations in <I>SPTB</I> gene account for about 20% patients.</P><P><B>Patient concerns:</B></P><P>A 65-year-old female had been diagnosed as hemolytic anemia 30 years ago, based on a history of persistent anemia and hyperbilirubinemia for several years. She received RBC transfusion several times and a cholecystectomy roughly 20 years ago before. Round, densely staining spherical-shaped erythrocytes (spherocytes) were frequently found on the PB smear. Numerous spherocytes were frequently found in the PB smears of symptomatic family members, her 3rd son and his 2 grandchildren.</P><P><B>Diagnosis:</B></P><P>One heterozygous mutation of <I>SPTB</I> was identified by targeted next-generation sequencing (NGS). The nonsense mutation, c.1956G>A (p.Trp652∗), in exon 13 was confirmed by Sanger sequencing and thus the proband was diagnosed with HS.</P><P><B>Interventions:</B></P><P>The proband underwent a splenectomy due to transfusion-refractory anemia and splenomegaly.</P><P><B>Outcomes:</B></P><P>After the splenectomy, her hemoglobin level improved to normal range (14.1 g/dL) and her bilirubin levels decreased dramatically (total bilirubin 1.9 mg/dL; direct bilirubin 0.6 mg/dL).</P><P><B>Lessons:</B></P><P>We suggest that NGS of causative genes could be a useful diagnostic tool for the genetically heterogeneous RBC membrane disorders, especially in cases with a mild or atypical clinical manifestation.</P>

Quantitative Determination of Absorption and First-Pass Metabolism of Apicidin, a Potent Histone Deacetylase Inhibitor

Shin, Beom Soo,Yoo, Sun Dong,Kim, Tae Hwan,Bulitta, Jurgen B.,Landersdorfer, Cornelia B.,Shin, Jeong Cheol,Choi, Jin Ho,Weon, Kwon-Yeon,Joo, Sang Hoon,Shin, Soyoung American Society for Pharmacology and Experimental 2014 Drug metabolism and disposition: the biological fa Vol.42 No.6

<P>Apicidin, a potential oral chemotherapeutic agent, possesses potent anti-histone-deacetylase activity. After oral administration, the total bioavailability of apicidin is known to be low (14.2%–19.3%). In the present study, we evaluated the factors contributing to the low bioavailability of apicidin by means of quantitative determination of absorption fraction and first-pass metabolism after oral administration. Apicidin was given to rats by five different routes: into the femoral vein, duodenum, superior mesenteric artery, portal vein, and carotid artery. Especially, the fraction absorbed (<I>F</I><SUB>X</SUB>) and the fraction that is not metabolized in the gut wall (<I>F</I><SUB>G</SUB>) were separated by injection of apicidin via superior mesenteric artery, which enables bypassing the permeability barrier. The <I>F</I><SUB>X</SUB> was 45.9% ± 9.7%, the <I>F</I><SUB>G</SUB> was 70.9% ± 8.1% and the hepatic bioavailability (<I>F</I><SUB>H</SUB>) was 70.6% ± 12.3%, while the pulmonary first-pass metabolism was minimal (<I>F</I><SUB>L</SUB> = 102.8% ± 7.4%), indicating that intestinal absorption was the rate-determining step for oral absorption of apicidin. The low <I>F</I><SUB>X</SUB> was further examined in terms of passive diffusion and transporter-mediated efflux by in vitro immobilized artificial membrane (IAM) chromatographic assay and in situ single-pass perfusion method, respectively. Although the passive diffusion potential of apicidin was high (98.01%) by the IAM assay, the in situ permeability was significantly enhanced by the presence of the P-glycoprotein (P-gp) inhibitor elacrider. These data suggest that the low bioavailability of apicidin was mainly attributed to the P-gp efflux consistent with the limited <I>F</I><SUB>X</SUB> measured in vivo experiment.</P>

Simultaneous analysis of acetylcarnitine, proline, hydroxyproline, citrulline, and arginine as potential plasma biomarkers to evaluate NSAIDs-induced gastric injury by liquid chromatography–tandem mass spectrometry

Shin, Soyoung,Jeong, Hyeon Myeong,Chung, Seung Eun,Kim, Tae Hwan,Thapa, Subindra Kazi,Lee, Da Young,Song, Chang Ho,Lim, Jun Young,Cho, Sang-Min,Nam, Kyu-Yeol,Kang, Won-Ho,Choi, Youn-Woong,Shin, Beom S Elsevier 2019 Journal of Pharmaceutical and Biomedical Analysis Vol.165 No.-

<P><B>Abstract</B></P> <P>Although major adverse effects associated with nonsteroidal anti-inflammatory drugs (NSAIDs) are gastric injury, assessment of NSAIDs-induced gastrointestinal adverse effects is mostly dependent on endoscopy due to the lack of plasma biomarkers. Several amino acids associated with collagenase activity and gastric mucosal mass have been suggested as plasma biomarker candidates for gastric injury. Therefore, this study aimed to develop a liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the plasma biomarker candidates, i.e., acetylcarnitine, proline, hydroxyproline, citrulline, and arginine and evaluate their potential as a biomarker for NSAIDs-induced gastric injury. The method utilized simple protein precipitation with methanol and D<SUB>4</SUB>-citrulline as an internal standard (IS). The assay resulted in the lower limit of quantification (LLOQ) of 0.1 μg/mL for acetylcarnitine and 1 μg/mL for proline, hydroxyproline, citrulline, and arginine in the surrogate blank plasma. The intra- and inter-day accuracy ranged 82.5–111.2% for acetylcarnitine, 95.4–103.3% for proline, 98.9–106.4% for hydroxyproline, 99.5–103.5% for citrulline, and 87.4–105.3% for arginine. The precision was within 6.17%, 3.63%, 6.20%, 6.31%, and 6.17% for acetylcarnitine, proline, hydroxyproline, citrulline, and arginine, respectively. The developed assay was successfully applied to monitor the changes of the plasma levels of the five amino acids in rats and Beagle dogs following repeated oral administrations of aceclofenac. In rats, plasma concentrations of proline, hydroxyproline, and citrulline were significantly reduced after 4 days of aceclofenac administration compared to the control group. In dogs, plasma concentrations of proline and citrulline were significantly decreased after 7 days of aceclofenac administration compared to those obtained after the first aceclofenac administration. These data indicate that plasma levels of proline, hydroxyproline, and citrulline may be used as quantitative biomarkers of NSAIDs-induced gastric damage. The present assay could also be utilized to monitor the changes of these amino acids as potential indicators for various physiological and pathophysiological conditions.</P> <P><B>Highlights</B></P> <P> <UL> <LI> LC–MS/MS analysis for simultaneous determination of 5 amino acids was developed. </LI> <LI> The assay was fully validated and applied to in vivo studies in rats and dogs. </LI> <LI> Proline, hydroxyproline, and citrulline levels were reduced by aceclofenac doses. </LI> <LI> The assay may be useful to monitor changes of the amino acid levels as biomarkers. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Complement 4 levels of a 4-year-old girl with angioedema

Shin, Soyoung,Lee, Yoon Tae,Lee, Kyung Yil,Park, Joonhong,Lee, Jae Ho,Yang, Eun Ae The Korean Pediatric Society 2020 Clinical and Experimental Pediatrics (CEP) Vol.63 No.1

Investigation of Terrain Slope Information and Tractor Driving Simulation on Reconstructed Ground

( Soyoung Shin ),( Eiji Inoue ),( Muneshi Mitsuoka ),( Takashi Okaysu ),( Yasmaru Hirai ),( Masami Matsui ) 한국농업기계학회 2018 한국농업기계학회 학술발표논문집 Vol.23 No.1

Despite the efforts of many researchers to improve the safety of agricultural machinery, fatalities have been caused by agricultural machinery annually. In Japan, about 50% of casualties caused by agricultural machinery are occurred by tractor rollover. In particular, the rollover of agricultural machinery causes major fatalities on the farm road. Agricultural machinery is easily rollover due to its high center of gravity when compared to other vehicles. In addition, when the implements are attached, the center of gravity moves according to the position of the implements, thereby exposing it to a greater risk of rollover. ROPS has been developed in an effort to reduce fatalities caused by tractor rollover accidents. The ROPS keeps the operator's space in case the tractor is overturned, preventing the operator from laying down from the tractor. Since 1997, ROPS has been mandatory for all riding tractors in Japan. Despite these efforts, however, there are fatalities caused by tractor rollover, and safety devices such as ROPS and seatbelts can not prevent fundamental accidents. In this study, we reconstruct the terrain images in front of the tractor using the stereo camera and analyze the slope of the reconstructed ground to predict the rollover accident of the tractor sensitive to the slope. The high center of gravity provides an opportunity to rollover in response to unexpected obstacles or steep slope. A stereo camera was used to record the frontal ground, and the obtained images were processed into a three-dimensional space through image processing in Matlab. The slope of the road surface on which the tractor is to be driven was identified, and the tractor maneuvering was identified through simulation of the motion model.

중학생의 성취 수준에 따른 탐구적 과학 글쓰기 (Science Writing Heuristic) 수업의 효과

신소영 ( Soyoung Shin ),최애란 (Aeran Choi),박종윤 ( Jong Yoon Park ) 한국과학교육학회 2013 한국과학교육학회지 Vol.33 No.5

The purpose of this study was to investigate the effects of the Science Writing Heuristic (SWH) approach on the students` summary writing, logical thinking and achievements for the course. Participants in this study were 132 female students from a girls` middle school. The SWH approach was used for two experimental classes and the typical teacher-centered instructional approach was used for two comparative classes. Summary writing test, logical thinking test (GALT) and achievement test for the course were administered before and after the instruction period. Results of this study indicated that the SWH approach was helpful for students in finding big ideas, understanding science concepts, developing logical thinking abilities and doing well in the course. This study also implied that the SWH approach was effective for the low achieving students.