• Title/Summary/Keyword: Fluorescent dye

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Protective effect of Caryophylli Flos on apoptosis caused by oxidative stress in HaCaT cells (HaCaT 세포의 산화 스트레스로 인한 세포자멸사에서 정향의 보호효과)

  • Park, Sook Jahr
    • The Korea Journal of Herbology
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    • v.36 no.5
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    • pp.93-99
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    • 2021
  • Objective : Caryophylli Flos has been used in Korean medicine to relieve vomiting and pains caused by chills that make fluid circulation difficult. This study was designed to investigate the protective effect of ethanol extract of Caryophylli Flos (CF) in hydrogen peroxide (H2O2)-induced apoptotic cell death in human keratinocyte HaCaT cells. Methods : CF was prepared by extracting 200 g of Caryophylli Flos in 2 L of ethanol for 48 h. Cell viability was measured by MTT assay, and the protein expression was monitored by Western blot analysis. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Reactive oxygen species (ROS) was measured using fluorescent dye, and reduced glutathione (GSH) was determined with a colorimetric commercial kit. Results : CF protected HaCaT cells from cell death caused by oxidative stress after H2O2 treatment. H2O2 amplified generation of ROS and induced depletion of GSH, whereas these changes in ROS and GSH were inhibited by GF treatment. In addition, H2O2 resulted in apoptosis as assessed by TUNEL assay and the expression of apoptosis regulator proteins. However, cells treated with CF showed a decrease in TUNEL-positive cells and restored the reduced expression of procaspase-9, -3 and PARP. Conclusion : This study showed cytoprotective effects of CF by anti-apoptotic activity while exerting antioxidative activity in H2O2-treated HaCaT cells. These results suggest that CF could be beneficial in skin damage caused by oxidative stress.

Assessing the Influence Radius of a Water Treatment System Installed in a Reservoir Using Tracer Experiment and 3D Numerical Simulation (추적자 실험 및 3차원 수치모의를 이용한 저수지 수처리 장치의 영향반경 평가)

  • Park, Hyung Seok;Lee, Eun Ju;Ji, Hyun Seo;Choi, Sun Hwa;Chun, Se Woong
    • Journal of Korean Society of Water Science and Technology
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    • v.26 no.6
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    • pp.3-12
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    • 2018
  • The objective of this study was to evaluate the radius of influence of effluent of water treatment system developed for the purpose of improvement of reservoir water quality using fluorescent dye (Rhodamine-WT) tracer experiment and 3-D numerical model. The tracer experiment was carried out in a medium-sized agricultural reservoir with a storage capacity of $227,000m^3$ and an average depth of 1.6 m. A guideline with a total length of 160 m was installed at intervals of 10 m in the horizontal direction from the discharge part, and a Rhodamine measurement sensor (YSI 6130, measurement range $0-200{\mu}g/L$) was used to measure concentration changes in time, distance, and depth. Experimental design was established in advance through Jet theory and the diffusion process was simulated using ELCOM, a three dimensional hydraulic dynamics model. As a result of the study, the direct effect radius of the jet emitted from the applied water treatment system was about 50-70 m, and the radius of physical effect by the advection diffusion was judged to be 100-120 m. The numerical simulations of effluent advection-diffusion of the water treatment system using ELCOM showed very similar results to those of the impact radius analysis using the tracer experiment and jet flow empirical equations. The results provide valuable information on the spatial extent of the water quality improvement devices installed in the reservoir and the facility layout design.

Nootkatol prevents ultraviolet radiation-induced photoaging via ORAI1 and TRPV1 inhibition in melanocytes and keratinocytes

  • Woo, Joo Han;Nam, Da Yeong;Kim, Hyun Jong;Hong, Phan Thi Lam;Kim, Woo Kyung;Nam, Joo Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.1
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    • pp.87-94
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    • 2021
  • Skin photoaging occurs due to chronic exposure to solar ultraviolet radiation (UV), the main factor contributing to extrinsic skin aging. Clinical signs of photoaging include the formation of deep, coarse skin wrinkles and hyperpigmentation. Although melanogenesis and skin wrinkling occur in different skin cells and have different underlying mechanisms, their initiation involves intracellular calcium signaling via calcium ion channels. The ORAI1 channel initiates melanogenesis in melanocytes, and the TRPV1 channel initiates MMP-1 production in keratinocytes in response to UV stimulation. We aimed to develop a drug that may simultaneously inhibit ORAI1 and TRPV1 activity to help prevent photoaging. We synthesized nootkatol, a chemical derivative of valencene. TRPV1 and ORAI1 activities were measured using the whole-cell patch-clamp technique. Intracellular calcium concentration [Ca2+]i was measured using calcium-sensitive fluorescent dye (Fura-2 AM). UV-induced melanin formation and MMP-1 production were quantified in B16F10 melanoma cells and HaCaT cells, respectively. Our results indicate that nootkatol (90 μM) reduced TRPV1 current by 94% ± 2% at -60 mV and ORAI1 current by 97% ± 1% at -120 mV. Intracellular calcium signaling was significantly inhibited by nootkatol in response to ORAI1 activation in human primary melanocytes (51.6% ± 0.98% at 100 μM). Additionally, UV-induced melanin synthesis was reduced by 76.38% ± 5.90% in B16F10 melanoma cells, and UV-induced MMP-1 production was reduced by 59.33% ± 1.49% in HaCaT cells. In conclusion, nootkatol inhibits both TRPV1 and ORAI1 to prevent photoaging, and targeting ion channels may be a promising strategy for preventing photoaging.

Protective effect of Citrus unshiu peel on the cadmium-induced apoptosis in HepG2 cells (카드뮴으로 유발한 간세포 자멸사에서 진피의 보호효과)

  • Noh, Gyu Pyo;Byun, Sung Hui;Lee, Jong Rok;Park, Sook Jahr;Kim, Sang Chan
    • The Korea Journal of Herbology
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    • v.36 no.1
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    • pp.41-49
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    • 2021
  • Objective : Citrus unshiu peel (Citri Unshius Pericarpium) has been prescribed to suppress coughing and phlegm in Korean medicine. In this study, the effect of ethanol extract of Citrus unshiu peel (CEE) on apoptosis was investigated using cadmium chloride (CdCl2) treated HepG2 cells. Methods : CEE was prepared by extracting 300 g of Citri Unshius Pericarpium in 3 L of ethanol for 72 h. Apoptosis was determined by the TUNEL assay. The mitochondrial membrane potential (MMP) was monitored using the membrane-permeable fluorescent dye Rh123. The expression level of each protein was monitored by Western blot analysis. Results : CEE protected HepG2 cells from apoptosis as determined by the TUNEL assay. A decrease in MMP was observed in cells exposed to cadmium, indicating that mitochondria are involved in the induction of apoptosis. However, CEE recovered the reduction in MMP caused by cadmium. In addition, decreased expression of B-cell lymphoma 2 (Bcl-2), procaspase, and poly(ADP-ribose) polymerase (PARP) by cadmium was increased by CEE. The anti-apoptotic effect of CEE was found to be associated with inhibition of JNK and p38 phosphorylation when examining the expression of phosphorylated MAPK by Western blot. Conclusion : This study showed that CEE exerted anti-apoptotic effects in cadmium-induced HepG2 cells by inhibiting the reduction of MMP and changes in the expression level of apoptotic proteins. These results suggest the potential for CEE to be used for heavy metal-induced liver damage.

Cytoprotective effect of the processed Gardeniae Fructus on oxidative stress-induced gastric epithelial cells (산화스트레스를 유발한 위 상피세포에서 수치 치자의 세포 보호 효과)

  • Jong Rok, Lee;Sang Chan, Kim;Sook Jahr, Park
    • The Korea Journal of Herbology
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    • v.38 no.1
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    • pp.21-30
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    • 2023
  • Objective : Gardeniae Fructus (GF) has bitter and cold nature. Thus, it has been traditionally prescribed in processed form roasted with ginger juice for patients with a weak stomach. This study investigated the effects of processed GF in tert-butyl hydroperoxide (tBHP)-treated gastric epithelial cells. Methods : Processed GF was made by applying 40% ginger juice or 10% ethanol for 24 h and then roasting at 150℃ for 5 minutes. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Mitochondrial membrane potential (MMP) was monitored by flow cytometry using the membrane permeable fluorescent dye Rh123. Protein expression was measured by Western blot analysis. Results : Cell viability was reduced by tBHP and restored by ethanol extract of GF (GFE). In the TUNEL assay, it was found that cell death by tBHP was due to apoptosis, and GFE had an anti-apoptotic effect. Processed GF roasted with ginger juice showed the best anti-apoptotic effect. Processed GF also inhibited MMP loss and restored tBHP-induced changes in expression levels of apoptosis-related proteins. Increased ROS production and GSH depletion after tBHP treatment were significantly reduced by processed GF. In addition, tBHP-induced activation of mitogen-activated protein kinase (MAPK) was inhibited by processed GF. Conclusion : These results demonstrate that the processed GF is able to protect gastric epithelial cells from oxidative stress-induced cell death with antiapoptotic and antioxidant activity. In addition, it shows that the processing of GF, which have been traditionally used for gastrointestinal protection, partially have scientific validity.

A new phantom to evaluate the tissue dissolution ability of endodontic irrigants and activating devices

  • Kimia Khoshroo ;Brinda Shah;Alexander Johnson ;John Baeten ;Katherine Barry;Mohammadreza Tahriri ;Mohamed S. Ibrahim;Lobat Tayebi
    • Restorative Dentistry and Endodontics
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    • v.45 no.4
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    • pp.45.1-45.8
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    • 2020
  • Objective: The aim of this study was to introduce a gelatin/bovine serum albumin (BSA) tissue standard, which provides dissolution properties identical to those of biological tissues. Further, the study evaluated whether the utilization of endodontic activating devices led to enhanced phantom dissolution rates. Materials and Methods: Bovine pulp tissue was obtained to determine a benchmark of tissue dissolution. The surface area and mass of samples were held constant while the ratio of gelatin and BSA were varied, ranging from 7.5% to 10% gelatin and 5% BSA. Each sample was placed in an individual test tube that was filled with an appropriate sodium hypochlorite solution for 1, 3, and 5 minutes, and then removed from the solution, blotted dry, and weighed again. The remaining tissue was calculated as the percent of initial tissue to determine the tissue dissolution rate. A radiopaque agent (sodium diatrizoate) and a fluorescent dye (methylene blue) were added to the phantom to allow easy quantification of phantom dissolution in a canal block model when activated using ultrasonic (EndoUltra) or sonic (EndoActivator) energy. Results: The 9% gelatin + 5% BSA phantom showed statistically equivalent dissolution to bovine pulp tissue at all time intervals. Furthermore, the EndoUltra yielded significantly more phantom dissolution in the canal block than the EndoActivator or syringe irrigation. Conclusions: Our phantom is comparable to biological tissue in terms of tissue dissolution and could be utilized for in vitro tests due to its injectability and detectability.

Metal Oxide Thin Film Transistor with Porous Silver Nanowire Top Gate Electrode for Label-Free Bio-Relevant Molecules Detection

  • Yu, Tae-Hui;Kim, Jeong-Hyeok;Sang, Byeong-In;Choe, Won-Guk;Hwang, Do-Gyeong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.268-268
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    • 2016
  • Chemical sensors have attracted much attention due to their various applications such as agriculture product, cosmetic and pharmaceutical components and clinical control. A conventional chemical and biological sensor is consists of fluorescent dye, optical light sources, and photodetector to quantify the extent of concentration. Such complicated system leads to rising cost and slow response time. Until now, the most contemporary thin film transistors (TFTs) are used in the field of flat panel display technology for switching device. Some papers have reported that an interesting alternative to flat panel display technology is chemical sensor technology. Recent advances in chemical detection study for using TFTs, benefits from overwhelming progress made in organic thin film transistors (OTFTs) electronic, have been studied alternative to current optical detection system. However numerous problems still remain especially the long-term stability and lack of reliability. On the other hand, the utilization of metal oxide transistor technology in chemical sensors is substantially promising owing to many advantages such as outstanding electrical performance, flexible device, and transparency. The top-gate structure transistor indicated long-term atmosphere stability and reliability because insulator layer is deposited on the top of semiconductor layer, as an effective mechanical and chemical protection. We report on the fabrication of InGaZnO TFTs with silver nanowire as the top gate electrode for the aim of chemical materials detection by monitoring change of electrical properties. We demonstrated that the improved sensitivity characteristics are related to the employment of a unique combination of nano materials. The silver nanowire top-gate InGaZnO TFTs used in this study features the following advantages: i) high sensitivity, ii) long-term stability in atmosphere and buffer solution iii) no necessary additional electrode and iv) simple fabrication process by spray.

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Effect of Hypoxia on the Signal Transduction of Apoptosis in Osteoblasts (저산소 상태에서 조골세포 고사의 신호전달 기전)

  • Park, Young-Joo;Oh, Soh-Taek;Kang, Kyung-Hwa;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.33 no.6 s.101
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    • pp.453-463
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    • 2003
  • Mammalian cell is critically dependent on a continuous supply of oxygen. Even brief periods of oxygen deprivation can result in profound cellular damage. The aim of this study was to examine the possible mechanism of apoptosis in response to hypoxia in MC3T3E1 osteoblasts. MC3T3El osteoblasts under hypoxic conditions ($2\%$ oxygen) resulted in apoptosis in a time-dependent manner, determined by DNA fragmentation assay and nuclear morphology, stained with fluorescent dye (Hoechst 33258) Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, suppressed the DNA ladder in response to hypoxia in a concentration dependent manner. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-l activity (YVADase) was detected. To confirm what caspases were involved in apoptosis, western blot analysis was performed using an anticaspase-3 or 6 antibody. The 17-kDa protein, that corresponds to the active products of caspase-3 and the 20-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged lysates, in which the full length forms of caspase-3 and 6 were evident. With a time course similar to caspase-3 and 6 activation, hypoxic stress also caused the cleavage of Lamin A, typical of caspase-6 activity. In addition, the hypoxic stress elicited the release of cytochrome c into the cytosol during apoptosis. These findings suggested that the activation of caspases accompanied by a cytochrome c release in response to hypoxia was involved in apoptotic cell death in MC3T3E1 osteoblasts.

A mark-release-recapture experiment with Anopheles sinensis in the northern part of Gyeonggi-do, Korea

  • Cho, Shin-Hyeong;Lee, Hyeong-Woo;Shin, E-Hyun;Lee, Hee-Il;Lee, Wook-Gyo;Kim, Chong-Han;Kim, Jong-Taek;Lee, Jong-soo;Lee, Won-Ja;Jong, Gi-Gon;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.139-148
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    • 2002
  • In order to study the range of flight and feeding activity of Anopheles sinensis, the dispersal experiment was conducted in Paju city, located in the northern part of Gyeonggi-do, Republic of Korea, during the period of 7th to 28th September 1998. Unfed females An. sinensis were collected in cowshed and released after being marked with fluorescent dye at 23:00 hours on the same day. Released female mosquitoes were recaptured everyday during 21 days using light traps, which were set at 10 sites in the cowsheds located 1, 3, 6, 9 and 12 km north-northwest and north-northeast and at 3 sites located 1, 6 and 9 km toward south-west from the release point. In addition, to study the longest flight distance in one night, we set the light traps at 16 and 20 km toward north-northeast from the release site. All the collected mosquitoes were placed on filter papers and observed on UV transilluminator after treatment with one drop of 100% ethanol. Out of 12,773 females of An. sinensis released, 194 marked females mosquitoes were recaptured, giving 1.52% recapture rate. Of 194, 72 mosquitoes (37 1%) were recaptured in light traps from three places set at 1 km from the release point, 57 mosquitoes (29.4%) from two places at 1-3 km, 41 mosquitoes (21.1%) from three places at 3-6 km, 20 mosquitoes (10.3%) from three places at 6-9 km, and 4 mosquitoes (2.1%) from two places at 9-12 km. Since 170 female mosquitoes (87.6%) out of 194 marked mosquitoes were captured within 6 km from the release point, this flight radius represents the main activity area. An. sinensis was found to be able to fly at least 12 km during one night.

Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification (Reverse transcription Loop-mediated isothermal amplification을 이용한 Soybean mosaic virus의 진단)

  • Lee, Yeong-Hoon;Bae, Dae-Hyeon;Kim, Bong-Sub;Yoon, Young-Nam;Bae, Soon-Do;Kim, Hyun-Joo;Mainali, Bishwo P.;Park, In-Hee;Lee, Su-Heon;Kang, Hang-Won
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.315-320
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    • 2015
  • Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.