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Neuroprotective effect of Coreopsis lanceolata extract against hydrogen-peroxide-induced oxidative stress in PC12 cells

  • Kyung Hye Seo;Hyung Don Kim;Jeong-Yong Park;Dong Hwi Kim;Seung-Eun Lee;Gwi Young Jang;Yun-Jeong Ji;Ji Yeon Lee
    • Korean Journal of Agricultural Science
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    • v.49 no.2
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    • pp.175-184
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    • 2022
  • The present study investigated the neuroprotective effects of Coreopsis lanceolate extract against hydrogen-peroxide (H2O2)-induced oxidative damage and cell death in pheochromocytoma 12 (PC12) cells. Reactive oxygen species (ROS), 2,2'-azinobis (3-ethylbebzothiazoloine-6-sulfonic acid) diammonium salt, and 1,1-diphenyl-2-picrrylhydrazyl radical scavenging activities, as well as the expression levels of proteins associated with oxidative damage and cell death were investigated. According to the results, C. lanceolate extract exhibited inhibitory activity against intracellular ROS generation and cell-damaging effects induced by hydroxyl radicals in a dose-dependent manner. Total phenolic and flavonoid contents were 22.3 mg·g-1 gallic acid equivalent and 16.2 mg·g-1 catechin equivalent, respectively. Additionally, a high-performance liquid chromatography (HPLC) assay based on the internal standard method used to detect phenolic compounds. The phenolic compounds identified in C. lanceolata extract contained (+)-catechin hydrate (5.0 ± 0.0 mg·g-1), ferulic acid (1.6 ± 0.0 mg·g-1), chlorogenic acid (1.5 ± 0.0 mg·g-1), caffeic acid (1.2 ± 0.0 mg·g-1), naringin (0.9 ± 0.0 mg·g-1), and p-coumaric acid (0.5 ± 0.0 mg·g-1). C. lanceolata extract attenuated pro-apoptotic Bax expression levels and enhanced the expression levels of anti-apoptotic Bcl-2, caspase-3, and caspase-9 proteins. Therefore, C. lanceolata is a potential source of materials with neuroprotective properties against neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases.

Purification and Characterization of Mitochondrial Mg2+-Independent Sphingomyelinase from Rat Brain

  • Jong Min Choi;Yongwei Piao;Kyong Hoon Ahn;Seok Kyun Kim;Jong Hoon Won;Jae Hong Lee;Ji Min Jang;In Chul Shin;Zhicheng Fu;Sung Yun Jung;Eui Man Jeong;Dae Kyong Kim
    • Molecules and Cells
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    • v.46 no.9
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    • pp.545-557
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    • 2023
  • Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg2+-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg2+, Mn2+, Ni2+, Cu2+, Zn2+, Fe2+, and Fe3+ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg2+-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

Effects of Guar Gum Addition on Optimal Properties of Fish Surimi Mixture for 3D Printing (구아검 첨가가 3D 프린팅을 위한 수리미 혼합물의 특성에 미치는 영향)

  • Hyun-Sik Na;Dong-Hyeon Kim;Ha-Young Lee;Sang-Min Lee;Hyeon-Ji Yu;Go-wun Seo;ChangHyeon Ko;Seon-Woo Park;Hyung-Wook Choi;Ye-Jin Choi;Mi Jeong Jo;Dong-Woo Kim;DongHyun Ahn
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.6
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    • pp.790-797
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    • 2023
  • Guar gum is an edible starch extracted from Cyamopsis tetragonolobus and used as an auxiliary ingredient to enhance the viscosity and adhesiveness of food and improve its emulsification stability. Here, characteristics of fish surimi mixture with various added amounts (0, 0.23, 0.45, 0.67, and 0.89 wt%) of guar gum were analyzed. In unheated surimi with guar gum, the pH decreased as the content increased. In the color values analysis, only the b* (yellowness) increased depending on the amount of added guar gum. The firmness increased with increasing guar gum content, and no significant difference in adhesiveness was observed. In heated surimi, a* (redness) and b* decreased in a content-dependent manner, and L* (lightness) was higher than that of unheated surimi. Regarding physical properties, the highest values of hardness, gumminess, and chewiness were observed in the heated surimi with 0.67 wt% of guar gum. However, no significant change was observed in springiness and cohesiveness. Results of sensory evaluation indicated that with higher guar gum content, only the hardness increased slightly, and significant differences were absent in the remaining parameters. Thus, surimi with 0.67 wt% of guar gum is considered to be optimal for 3D printing.

Antiproliferative Effect of Extracts, Fractions and Compound from Vitex rotundifolia on Human Cancer Cells (순비기 나무(Vitex rotundifolia) 추출.분획물 및 화합물의 인체 암세포 증식억제 효과)

  • Kim, You-Ah;Lee, Jung-Im;Kim, Hae-Jin;Kong, Chang-Suk;Nam, Taek-Jeong;Seo, Young-Wan
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.180-186
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    • 2009
  • Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.

Priming Effect of Endotoxin in Human Alveolar Macrophage (사람 폐포대식세포에서 내독소의 Priming 효과)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.46-53
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    • 1996
  • Background: Endotoxin or lipopolysaccharide(LPS) can prime phagocytic cells such as polymorphonuclear leukocytes, monocytes or animal peritoneal macrophages to generate increased amounts of secretory products such as oxygen free radicals and tumor necrosis factor, which play an important role in developing adult respiratory distress syndrome in gram negative sepsis. Human alveolar macrophages(HAM) are continuously exposed to various stimuli inhaled into the alveoli, and the response to LPS might be different in HAM. Therefore, we investigated the effect of LPS pre-exposure on HAM adhered to plastic surface and A549 cell(type II human alveolar epithelial cell line) monolayer. Methods: HAM were isolated from bronchoalveolar lavage fluid from normal lung of the patients with localized lung cancer and esophageal cancer. LPS was exposed to HAM for 2hrs before or after adherence to plastic surface of 24-well Linbro plate and A549 cell monolayer. And then HAM was stimulated with PMA(phorbol myristate acetate) or fMLP(N-formyl-methionylleucyl-phenylalanine). The amount of hydrogen peroxide($H_2O_2$) production in the supernatant was measured on the principle of peroxidase-dependent oxidation of phenol red by hydrogen peroxide. Results: LPS pre-exposure could not enhance $H_2O_2$ production in neither HAM adhered to plastic surface nor one to A549 cell monolayer. But LPS even in the absence of PMA or fMLP stimulation directly increased $H_2O_2$ release in HAM if added after the adherence to A549 cell monolayer. Conclusion: Endotoxin does not prime HAM, but may directly activate HAM adhered to alveolar epithelial cells. Further investagation will be necessary.

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Anti-Stress Effect of Punica granatum L. Extract against Sleep Deprivation-Induced Impairment (석류 열수 추출물의 수면박탈을 유도한 Rat 모델에서의 항스트레스 효과)

  • Na, Ju-Ryun;Kim, Sunoh;Jo, Ara;Bae, Donghyuck;Oh, Kyo-Nyeo;Kim, Yong Jae;Lee, Yoo-Hyun;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1533-1543
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    • 2016
  • The anti-stress effects of Punica granatum L. (family Lythraceae, PG) on $H_2O_2$/corticosterone (CORT)-induced stress in cells and sleep-deprived rats were investigated. The PG extract showed neuroprotective effects in SH-SY5Y cells against $H_2O_2$/CORT-induced stress. Sleep deprivation led to behavioral, hormonal, and biochemical alterations in the animal model. The effects of P. granatum on physiological, behavioral, and biochemical parameters aggravated by sleep deprivation were investigated. Sleep deprivation impaired physiological (survival, body weight, and drowsiness scores) and behavioral (rotarod, passive avoidance, hot hyperalgesia, and Y maze) parameters as well as biochemical factors (cortisol, serotonin, dopamine, testosterone, and growth factor I contents in serum). These parameters were significantly recovered by PG extract in a concentration-dependent manner. The PG extract also enhanced catalase, superoxide dismutase, and non-enzymatic antioxidative activities such as glutathione compared to sleep-deprived rats. On the basis of these results, our findings suggest that Punica granatum prevents impairment of body functions induced by sleep deprivation and related oxidative damage.

Postprandial hypoglycemic effects of mulberry twig and root bark in vivo and in vitro (In vivo와 in vitro에서 상지 및 상백피 에탄올추출물이 식후 혈당 상승 억제 조절에 미치는 영향)

  • Park, Soo Yeon;Jin, Bo Ra;Lee, Yu Rim;Kim, You Jin;Park, Jeong Bin;Jeon, Young Hee;Choi, Sang Won;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.49 no.1
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    • pp.18-27
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    • 2016
  • Purpose: Our previous study demonstrated the hypoglycemic effects of mulberry (Morus alba L.) leaf and the underlying mechanisms. Here we explored the potency of mulberry twigs (TW) and root barks (RB) in postprandial hypoglycemic effects in vitro and in vivo. Methods: The major components of TW and RB were determined by high performance liquid chromatography (HPLC). Alpha-glucosidase inhibition and glucose/fructose uptake inhibition in Caco-2 cells were determined for TW, RB, and their major components, followed by an oral sugar tolerance test (OSTT) in streptozotocin-induced diabetic rats. Male Wistar rats were fed a high-fat diet for 2 weeks and then a single dose of streptozotocin (35 mg/kg B.W) was administered by intraperitoneal injection. Rats with fasting blood glucose levels above 126 mg/dL were randomly divided into 5 groups (n = 8/group) for the following treatments by gavage for 4 weeks: vehicle (normal control and diabetic control), 200 mg/kg B.W of TW or RB or 100 mg/kg B.W of oxyresveratrol (OXY). Results: OXY and mulberroside A were identified as the major components of TW and OXY, mongolicin, and kuwanon H for RB. A significant inhibitory activity on ${\alpha}-glucosidase$ was found for TW, RB, and OXY (p = 0.0099). There was a dose-dependent inhibition of TW and RB on the intestinal sugar uptakes in Caco-2 cells, showing a greater impact on fructose compared to glucose. The OSTT showed that TW and RB significantly delayed time to maximal concentration (p = 0.0088) and decreased maximal concentration (p = 0.0043) compared to the control group. Conclusion: These results suggest that TW and RB may have a postprandial hypoglycemic effect, particularly in the case of high fructose or sucrose intake. OXY was suggested as a contributor to the hypoglycemic effect of TW and RB. Further studies are needed for the systemic effect of TW and RB in circulation.

Seasonal variation of physico-chemical factors and size-fractionated phytoplankton biomass at Ulsan seaport of East Sea in Korea (동해 울산항에서 이화학적 환경요인 및 크기그룹별 식물플랑크톤 생체량의 계절적 변동)

  • Kwon, Oh Youn;Kang, Jung-Hoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.11
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    • pp.6008-6014
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    • 2013
  • This study aimed to understand seasonal variation of physico-chemical factors and biomass of size-fractionated phytoplankton at Ulsan seaport during the period from February 2007 to November 2009. Water temperature, salinity, dissolved oxygen (DO), pH, chemical oxygen demand (COD) and total suspended solid (TSS) varied in the range of 8.94-$24.26^{\circ}C$, 25.06-34.54 psu, 4.30-10.73 mg/L, 7.97-8.53, 0.66-40.70 mg/L and 57.4-103.3 mg/L, respectively. These factors showed no clear spatial variation unlike spatial pattern of inorganic nutrients and total chlorophyll-a (chl-a) concentration as biomass. Concentration of phosphate, nitrate and silicate ranged from 0.01 to 3.03 ${\mu}M$, 0.05 to 21.62 ${\mu}M$, and 0.01 to 27.82 ${\mu}M$, respectively, with 2 times higher concentration at inner stations than that at outer stations during the study period. Within the range of total chl-a concentration (0.36-7.11 ${\mu}gL^{-1}$), higher concentration (avg. 1.88 ${\mu}gL^{-1}$) of total chl-a were observed at inner stations compared to that (avg. 0.90 ${\mu}gL^{-1}$) at outer stations. Micro-sized phytoplankton dominated total biomass of phytoplankton in spring (34.0-81.2%), summer (35.1-65.6%) and winter (3.9-62.0%). Nano- and pico-sized phytoplankton contributed 58.2-74.5% and 22.4-38.2% to total biomass of phytoplankton in autumn, respectively. However, contribution in biomass of size-fractionated phytoplankton to total phytoplankton biomass showed no clear difference between inner and outer stations. Consequently, these results indicated that spatio-temporal distribution of phytoplankton biomass at Ulsan seaport was dominated by micro-phytoplankton (avg. 52.3%) during the study period except autumn, which was closely dependent on the concentration of inorganic nutrients (p<0.05).

17 beta-Estradiol Increases Peak of $\textrm{Ca}^{2+}$ Current in Mouse Early Embryo (에스트로겐이 생쥐 초기배의 $\textrm{Ca}^{2+}$ 전류에 미치는 영향)

  • 강다원;신용원;김은심;홍성근;한재희
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.79-89
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    • 2001
  • Steroid hormones control the expression of many cellular regulators, and a role thor estrogen in mouse oocytes has been well documented. The preovulatory $E_2$increment is generally accepted as the endocrine process regulating induction of in vivo oocyte maturation To address whether the activity of the T-type $Ca^{2+}$ channel is altered by 17 beta-estradiol ( $E_2$), we examined the actions of $E_2$on the calcium channel of mouse oocytes and early embryos. Oocrtes were collected from the oviduct of mice treated with pregnant mare's serum gonadotropin (PMSG) and human choronic gonadotropin (hCG). Whole cell voltage clamp technique and confocal microscopy were used to examine that $E_2$increase intracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{i}$ ) via voltage dependent $Ca^{2+}$ channel (VDC) and estrogen receptor (FSR), and $E_2$concentration by the use of radioimmunoassay (RIA) were examined in mouse. The results obtained were as follows: The peak of $Ca^{2+}$ current induced by $E_2$increased 122% to 1.50$\pm$0.03 nA from 1.23$\pm$0.21 nA (n=15) in the presence of 5 mM extracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{o}$ ). The increased $Ca^{2+}$ current was temporally associated with $Ca^{2+}$ transients. The intracellular $Ca^{2+}$ level increased 207%~30 s following the addition of 1${\mu}{\textrm}{m}$ $E_2$(relative fluorescence intensity: 836.4$\pm$131.2 for control, n=10, 1736.4$\pm$192.0 in the presence of $E_2$, n=10). $E_2$increased amplitude of $Ca^{2+}$ current and [C $a^{2+}$]$_{i}$ . $E_2$-induced $Ca^{2+}$ current and $E_2$concentration in blood were showed difference on the stage of embryo. These results suggest that $E_2$modulate $Ca^{2+}$ channel to increase $Ca^{2+}$ influx.$Ca^{2+}$ influx.

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How Skin Care Ingredient Concentrations Can Modulate the Effect of polyols and Oils on Skin Moisturization and Skin Surface Roughness (화장품 원료 중 폴리올, 오일 농도에 따른 피부 보습과 피부 표면 거칠기의 변화)

  • Nam, Gae-Won;Kim, Seung-Hun;Kim, Eun-Joo;Kim, Jin-Han;Chae, Byung-Guen;Lee, Hae-Kwang;Moon, Seong-Joon;Kang, Hak-Hee;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.337-342
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    • 2005
  • The aim of this study was to evaluate the influence of different skin care ingredient concentrations on the effect of polyols and oils on the human skin moisturization and skin surface roughness. Polyols and oils were essential ingredients to make a skin care formulation. But these were still not understood how much concentration(s) were tested on human skin in the aspect of efficacy and sensory. We studied to examine various concentrations of ingredient by cosmetic companies using noninvasive methods. Polyols were composed of glycerol and butylene glycol (BG) as 1:1 ratio, and oils were hydrogenated polydecene, cetyl ethylhexanoate and pentaerythrityl tetraethylhexanoate (PTO(R), Stearinerie Dubois Fils Co., France) as 1:1:1 ratio. All compounds were tested $0{\sim}27%dml$ Polyols and $0{\sim}35%dml$ oils in O/W emulsions. We investigated the effect of water contents and the effect of stratum corneum roughness in forearm skin after application of compounds. Water contents of the skin measured by skin capacitance and skin surface roughness measured visual scoring of skin surface biopsy through the scanning electron microscopy. Water contents of the skin were highly related to amount of polyols (to 20%) and oils (to 12%). Correlation coefficients were 0.971 and 0.985 respectively (p<0.01), 2 h after application. Skin surface roughness was positively correlated with polyol contents in concentration dependent manner, and depend on oils up to 6%. The ratio of coefficient was 2.5 to 1 (polyol to oils) by regression analysis. Further studies will be conducted with other ingredients such as surfactants, lipids and aqueous materials, and with ether methods for noninvasive measurement.