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Effects of Dangkwisoo-San, Ginger and Curcumin on Transient Receptor Potential Melastatin 7 Channels (당귀수산, 생강, 커큐민의 대사성 질환과 관련된 일과성 수용체 전압 이온통로조절에 관한 연구)

  • Kim, Byung Joo
    • Journal of Korean Medicine for Obesity Research
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    • v.18 no.1
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    • pp.10-18
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    • 2018
  • Objectives: Metabolic syndrome is correlated with increased cardiovascular risk and characterized by several factors, including visceral obesity, hypertension, insulin resistance, and dyslipidemia. Several members of a large family of nonselective cation entry channels, e.g., transient receptor potential (TRP) melastatin 7 (TRPM7) channels have been associated with the development of cardiovascular diseases. The purpose of this study was to investigate the effects of Dangkwisoo-san, ginger and curcumin on TRPM7 channel. Methods: Human embryonic kidney (HEK) 293 cells stably transfected with the TRPM7 expression vectors were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, $5{\mu}g/mL$ blasticidin, and 0.4 mg/mL zeocin in a humidified 20% $O_2$/10% $CO_2$ atmosphere at $37^{\circ}C$. Whole-cell patch clamp recordings were obtained using an Axopatch 700B amplifier and pClamp v.10.4 software, and signals were digitalized at 5 kHz using Digidata 1422A. Results: Dangkwisoo-san extract (100, 200, 300, 400, and $500{\mu}g/mL$) inhibited the outward and inward TRPM7 whole-cell currents at dose dependent manner and the half maximal inhibitory concentration $(IC)_{50}$ of Dangkwisoo-san was $218.3{\mu}g/mL$. Also, ginger extract (100, 200, 300, 400, and $500{\mu}g/mL$) inhibited the outward and inward of TRPM7 whole-cell currents in a dose dependent manner and the $IC_{50}$ of ginger was $877.2{\mu}g/mL$. However, curcumin had no effects on TRPM7 whole-cell currents. Conclusions: These results suggest that both Dangkwisoo-san and ginger have good roles to inhibit the TRPM7 channel, suggesting that Dangkwisoo-san and ginger are considered one of the candidate agents for the treatment of metabolic syndrome such as cardiovascular disease.

Improving the Safety of Mesenchymal Stem Cell-Based Ex Vivo Therapy Using Herpes Simplex Virus Thymidine Kinase

  • Bashyal, Narayan;Lee, Tae-Young;Chang, Da-Young;Jung, Jin-Hwa;Kim, Min Gyeong;Acharya, Rakshya;Kim, Sung-Soo;Oh, Il-Hoan;Suh-Kim, Haeyoung
    • Molecules and Cells
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    • v.45 no.7
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    • pp.479-494
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    • 2022
  • Human mesenchymal stem cells (MSCs) are multipotent stem cells that have been intensively studied as therapeutic tools for a variety of disorders. To enhance the efficacy of MSCs, therapeutic genes are introduced using retroviral and lentiviral vectors. However, serious adverse events (SAEs) such as tumorigenesis can be induced by insertional mutagenesis. We generated lentiviral vectors encoding the wild-type herpes simplex virus thymidine kinase (HSV-TK) gene and a gene containing a point mutation that results in an alanine to histidine substitution at residue 168 (TK(A168H)) and transduced expression in MSCs (MSC-TK and MSC-TK(A168H)). Transduction of lentiviral vectors encoding the TK(A168H) mutant did not alter the proliferation capacity, mesodermal differentiation potential, or surface antigenicity of MSCs. The MSC-TK(A168H) cells were genetically stable, as shown by karyotyping. MSC-TK(A168H) responded to ganciclovir (GCV) with an half maximal inhibitory concentration (IC50) value 10-fold less than that of MSC-TK. Because MSC-TK(A168H) cells were found to be non-tumorigenic, a U87-TK(A168H) subcutaneous tumor was used as a SAE-like condition and we evaluated the effect of valganciclovir (vGCV), an oral prodrug for GCV. U87-TK(A168H) tumors were more efficiently ablated by 200 mg/kg vGCV than U87-TK tumors. These results indicate that MSC-TK(A168H) cells appear to be pre-clinically safe for therapeutic use. We propose that genetic modification with HSV-TK(A168H) makes allogeneic MSC-based ex vivo therapy safer by eliminating transplanted cells during SAEs such as uncontrolled cell proliferation.

A rare ginsenoside compound K (CK) induces apoptosis for breast cancer cells

  • Seun Eui Kim;Myoung-Hoon Lee;Hye-Myoung Jang;Wan-Taek Im;Joontaik Lee;Sang-Hwan Kim;Gwang Joo Jeon
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.3
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    • pp.167-176
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    • 2023
  • Background: A breast cancer is the second leading cause of cancer death in women worldwide and among different types of breast cancers, triple-negative breast cancer (TNBC) has a poor prognosis. Methods: We investigated the potential of ginsenoside compound K (CK), an active ingredient in the bio-transformed ginsenoside, to be used as a therapeutic ingredient by examining the effects of CK on cell proliferation, apoptosis, and cancer-related gene expressions in breast cancer cells. Results: From the results of treating MCF-7, an ER and PR-positive breast cancer cells, and MDA-MB-231 (TNBC) with CK at a concentration of 0-100 µM, the half maximal inhibitory concentration (IC50) values for each cell were 52.17 µM and 29.88 µM, respectively. And also, it was confirmed that cell migration was inhibited above the IC50 concentration. In addition, fluorescence analysis of Apoptosis/Necrosis showed that CK induced apoptosis rather than necrosis of breast cancer cells. Through qPCR, it was confirmed that the expression of genes related to apoptosis and cell cycle arrest was increased in CK-treated breast cancer cells, and it acted more effectively on TNBC. However, the expression of genes related to tumor invasion and metastasis is also increased, so it is necessary to consider the timing of application of CK as a potential therapeutic anticancer compound. Conclusions: CK showed a stronger inhibitory effect in TNBC with poor prognosis but considering the high tumor invasion and metastasis-related gene expression, the timing of application of CK should be considered.

The Effect of Environmental Factors on the Advent of Chattonella (Raphidophyceae) in Yeosu Coastal Waters, Korea, and the Effect of Nutrients on the Growth of Chattonella (여수 연안해역에서 침편모조류 Chattonella속 출현환경 및 영양염에 대한 성장특성)

  • Noh, Il-Hyeon;Oh, Seok-Jin;Shin, Hyeon-Ho;Kang, In-Seok;Yoon, Yang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.4
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    • pp.362-372
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    • 2010
  • In order to understand what leads to the appearance of harmful Chattonella algae in the Yeosu coastal waters of Korea, we measured environmental parameters every week at one station from May to November, 2006, and April to October, 2007. Four species of Chattonella appeared during the monitoring period: C. antiqua, C. globosa, C. marina and C. ovata. The range of water temperature and salinity were $15.0-27.9^{\circ}C$ and 17.6~33.0 psu, respectively, when Chattonella appeared, and their maximum cell density (4,840 cells/L) was at $27.1^{\circ}C$ and 33.0 psu. During the monitoring periods, the range of dissolved inorganic nitrogen (DIN), phosphate (DIP) and chlorophyll $\alpha$ (Chl-$\alpha$) concentrations in surface waters were $1.20-52.23\;{\mu}M$ ($8.59{\pm}8.97\;{\mu}M$), $0.03-1.56\;{\mu}M$ ($0.47{\pm}0.31\;{\mu}M$) and $0.45-31.12\;{\mu}g/L$ ($3.58{\pm}4.77\;{\mu}g/L$), respectively. Chattonella occurred at low cell density when the Chl-$\alpha$ concentration increased because of supplied nutrients, whereas their cell density increased during the periods of rapid decrease in Chl-$\alpha$. The results of growth experiments based on batch culture showed that the half saturation constant ($K_s$) of C. antiqua on ammonium (${NH_4}^-$), nitrate (${NO_3}^-$) and phosphate (${PO_4}^{2-}$) were $3.89{\mu}M$, $5.01\;{\mu}M$ and $0.63\;{\mu}M$, respectively. These Ks values are higher than those reported for diatoms and other flagellates at the DIP concentration (average $0.47{\mu}M$) of Yeosu coastal waters. Although the maximum specific growth rate (${\mu}_{max}$) of C. antiqua was lower than diatoms, it was higher than those of other flagellates. Therefore, our results indicate that the DIP level in the study area was too low to support Chattonella blooms, although Chattonella species have physiological characteristics that enable them to grow more rapidly than other flagellates when nutrient levels are higher than their $K_s$.

Endothelial Cell Proliferation and Vascular Endothelial Growth Factor Expression in Primary Colorectal Cancer and Corresponding Liver Metastases

  • Raluca, Balica Amalia;Cimpean, Anca Maria;Cioca, Andreea;Cretu, Octavian;Mederle, Ovidiu;Ciolofan, Alexandru;Gaje, Pusa;Raica, Marius
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4549-4553
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    • 2015
  • Background: Colorectal carcinoma (CRC) is one of the major causes of cancer death worldwide. Data from the literature indicate differences between the proliferation rate of endothelial cells relative to the morphology growth type, possibly due to origin of specimens (autopsy material, surgery fragments) or quantification methods. Vascular endothelial growth factor (VEGF) is a factor that stimulates the proliferation of endothelial cells. It is expressed in more than 90% of cases of metastatic CRC. Aim: The aim of this study was to evaluate the endothelial cell proliferation and VEGF expression in primary tumors and corresponding liver metastases. Materials and Methods: Our study included 24 recent biopsies of primary tumors and corresponding liver metastases of CRC cases. CD34/Ki67 double immunostaining and RNA scope assay for VEGF were performed. Results: In the primary tumors analysis of VEGFmRNA expression indicated no significant correlation with differentiation grade, proliferative and non-proliferative vessels in the intratumoral and peritumoral areas. In contrast, in the corresponding liver metastases, VEGFmRNA expression significantly correlated with the total number of non-proliferative vessels and total number of vessels. CD34/Ki67 double immunostaining in the cases with poorly differentiated carcinoma indicated a high number of proliferating endothelial cells in the peritumoral area and a low number in the intratumoral area for the primary tumor. Moderately differentiated carcinomas of colon showed no proliferating endothelial cells in the intratumoral area in half of the cases included in the study, for both, primary tumor and liver metastasis. In well differentiated CRCs, in primary tumors, a high proliferation rate of endothelial cells in the intratumoral area and a lower proliferation rate in the peritumoral area were found. A low value was found in corresponding liver metastasis. Conclusions: The absence of proliferative endothelial cells in half of the cases for the primary tumors and liver metastases in moderately differentiated carcinoma suggest a vascular mimicry phenomenon. The mismatch between the total number of vessels and endothelial proliferation in primary tumors indicate that a functional vascular network is already formed or the existence of some mechanisms influenced by other angiogenic factors.

Synthesis and Crystal Structure of Zinc Iodide in the Sodalite Cavities of Zeolite A (LTA)

  • Kim, Seok-Han;Park, Man;Son, Young-Ja;Lee, Hyung-Joo;Jeong, Gyo-Cheol;Bae, Myung-Nam;Lim, Woo-Taik
    • Bulletin of the Korean Chemical Society
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    • v.28 no.4
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    • pp.567-573
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    • 2007
  • The crystal structure of ZnI2 molecule synthesized in zeolite A (LTA) has been studied by single-crystal X-ray diffraction techniques. A single crystal of |Zn6|[Si12Al12O48]-LTA, synthesized by the dynamic ion-exchange of |Na12|[Si12Al12O48]-LTA with aqueous 0.05 M Zn(NO3)2 and washed with deionized water, was placed in a stream of flowing 0.05 M KI in CH3OH at 294 K for four days. The resulting crystal structure of the product (|K6Zn3(KI)3(ZnI2)0.5|[Si12Al12O48]-LTA, a = 12.1690(10) A) was determined at 294 K by single-crystal X-ray diffraction in the space group Pm3m. It was refined with all measured reflections to the final error index R1 = 0.078 for 431 reflections which Fo > 4σ (Fo). At four crystallographically distinct positions, 3.5 Zn2+ and nine K+ ions per unit cell are found: three Zn2+ and five K+ ions lie on the 3-fold axes opposite 6-rings in the large cavity, two K+ ions are off the plane of the 8-rings, two K+ ions are recessed deeply off the plane of the 8-rings, and the remaining a half Zn2+ ion lie on the 3-fold axes opposite 6-rings in the sodalite cavity. A half Zn2+ ion and an I- ion per unit cell are found in the sodalite units, indicating the formation of a ZnI2 molecule in 50% of the sodalite cavities. Each ZnI2 (Zn-I = 3.35(5) A) is held in place by the coordination of its one Zn2+ ion to the zeolite framework oxygens and by the coordination of its two I- ions to K+ ions through 6-rings (I-K = 3.33(8) A). Three additional I- ions per unit cell are found opposite a 4-ring in the large cavity and form a K3I2+ and two K2ZnI3+ ionic clusters, respectively.

Effects of PtMn composition on carbon supported PtMn catalysts for PEMFC (Mn조성비(組成比)가 PEMFC용(用) Pt/C 전극촉매(電極觸媒) 특성(特性)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Yoo, Sung-Yeol;Kang, Suk-Min;Lee, Jin-A;Rhee, Choong-Kyun;Ryu, Ho-Jin
    • Resources Recycling
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    • v.21 no.2
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    • pp.34-40
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    • 2012
  • $Pt_{10}$/C, $Pt_9Mn_1$/C, $Pt_7Mn_3$/C electrocatalysts for Polymer Electrolyte Membrane Fuel Cells(PEMFCs) were synthesized by reduction with HCHO and their activity as a oxygen reduction reaction(ORR) was examined at half cell. The electrochemical oxygen reduction reaction(ORR) was studied by using a glaasy carbon electrode through cyclic voltammetric curves(CV) in a 1 M $H_2SO_4$ solution. The ORR activities of $Pt_9Mn_1$/C were higher than $Pt_{10}$/C, $Pt_7Mn_3$/C. Also potential-current curves of $Pt_9Mn_1$/C at 0.9, 0.8, 0.7, 0.6V for 5minutes respectively were higher than $Pt_{10}$/C, $Pt_7Mn_3$/C. Physical characterization was made by using x-ray diffraction(XRD) and transmission electron microscope(TEM). The TEM images of $Pt_9Mn_1$/C, $Pt_{10}$/C catalysts showed homogenous particle distribution with particle size of about 2.7 nm, 3 nm respectively and then the XRD results showed that the crystalline structure of the synthesized catalysts are seen FCC structure.

Effect of Citric Acid and Tetracycline HCl Root Conditioning on rhBMP-2 on Human Periodontal Ligament Fibroblast and Osteoblast cell (구연산과 테트라싸이클린으로 처리한 치근면에서 rhBMP-2가 치주인대섬유아세포와 골아세포의 활성에 미치는 영향)

  • Shim, Jung-Min;Han, Soo-Boo;Seol, Yang-Jo;Lee, Yong-Moo;Kim, Kyeong-Hwa;Kye, Seung-Beom;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.21-41
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    • 2001
  • The goal of Periodontal treatment is predictable periodontal regeneration. But until now, many products including GTR materials and growth factors are beyond of complete regeneration. BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. BMP can promote periodontal regeneration by their ability to stimulate new bone and new cementum formation. But little is known about optimal conditions required for the application. Root conditioning is used for bioacive root change so altered root surface provides a substrate that promotes chemotaxis, migration and attachment of peridontal cells encouraging connective attachment to the denuded root surface. The aim of this study is to investigate whether the acid conditioning change effect of rhBMP-2 on human periodontal ligament cell and osteoblast cell line. 288 periodontally involved root dentin slices are divided into 6 groups, each 48, 1)control, 2)treated with BMP, 3)treated with citric acid 4)treated with citric acid+BMP 5)treated with tetracycline 6)treated with TC+BMP. Each group was devided half, so 12 root dentin slices were seeded with periodontal ligament cells and 12 were seeded with osteoblasts. At day 2 and 7, cell number, protein assay, ALP activitiy was measured. To investigate morphology of cultured cells, SEM was employed. Statistical analysis was performed with SPSS 8.0 either t-test or ANOVA test. The results are ; Protein assay and cell number was slightly decreased in CA+BMP group compared to Ca group but it was not statistically significant and ALP activity was much more increased in CA+BMP group compared to CA group so there was no statistically significance between BMP and CA+BMP group and statistically significant compared to control group. Cell number and protein assay was slightly increased in TC group and ALP activity was much less the BMP group and CA group. Cell number and protein and ALP activity was not much increased in TC+BMP group. TC group and TC+BMP group showed cell morphology change in SEM. This results suggested that application of root surface with citric acid before BMP treatment might give better result in periodontal regeneration.

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Protective Effects of Kaempferol and Quercetin on Oxidative Stress in CPAE Cell (CPAE 세포를 이용한 Kaempferol과 Quercetin의 산화스트레스 극복효과)

  • Park, Shin-Young
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.395-401
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    • 2008
  • Flavonoids are ubiquitous substances in fruits and vegetables. A main subgroup of the flavonoids are the flavonols, of which quercetin and kaempferol are the major representatives in foods. They are used in food supplements at high doses, because of their preventive effects on degenerative diseases. The aim of this study was to determine the combined and separate effects of kaempferol and quercetin on oxidative stress in cow pulmonary artery endothelium (CPAE) cells over a broad concentration range. The results demonstrate that cell viability was greatly increased in kaempferol and quercetin treated cells whether $H_{2}O_{2}$-treated or not. Cell viability also increased when treated with flavonols in the absence of oxidative stress. Both preincubation and simultaneous incubation with kaempferol and quercetin protected against the loss of cell viability induced by 1mM $H_{2}O_{2}(5h)$. Protective effects of flavonols against oxidative stress were shown to depend on the treated flavonol concentrations. No protective effect was shown under low concentration treatment and cell viability increased 1.6 times at $200{\mu}M$ relative to the control group. At the highest flavonol concentration of $300{\mu}M$, the increased cell viability by flavonol treatment was decreased to almost half of the maximum values. Combined treatments with kaempferol and quercetin showed more protective effects against oxidative stress by $H_{2}O_{2}$ than the separate effects of each flavonol. In conclusion, the protective effect of kaempferol and quercetin against oxidative stress was very pronounced but high concentrations of flavonols can also induce cell cytotoxicity.

Effects of Taurine on Glutamate-induced Neurotoxicity and Interleukin-6 mRNA Expression in Astrocytes

  • Yang, Seong-Chil;Baek, Su-Yeon;Choe, In-Pyo;Lee, Chang-Jung
    • Animal cells and systems
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    • v.1 no.3
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    • pp.467-473
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    • 1997
  • Taurine (2-aminoethanesulfonic acid), one of bioactive amino acid in the mammalian brain, is known to exert inhibitory effects on neurons via GABA receptor. In the present study, we examined effects of taurine on glutamateinduced neurotoxicity on hippocampal neuron cell culture using cell counting method and lactate dehydrogenase (LDH) assay. After 10 d of culture, cells were stimulated with appropriate drugs. Only 43% of cultured neuronal cells survived at one day after stimulation with 500 uM L-glutamate for 10 min. Survival rate was enhanced by 82% in the presence of 10 mM taurine. LDH activity from the culture supernatant incubated with a combination of L-glutamate and taurine was less than half of that with L-glutamate alone. In the next series of experiments, interleukin-6 (IL-6) mRNA expression in cultured astrocytes was investigated using reverse tanscription-PCR (RT-PCR). IL-6 mRNA was detected in the astrocytes stimulated with L-glutamate in a dose-dependent manner, while not detected in the unstimulated control astrocytes. The expression of IL-6 mRNA caused by 10 mM glutamate was inhibited by taurine, but not by GABA. These findings demonstrated a neuroprotective action of taurine against glutamate-induced toxicity.

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