• Title/Summary/Keyword: AMP

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Characterization and Functional Analysis of Obox4 during Oocyte Maturation by RNA Interference (생쥐의 난소와 난자에서의 Obox4의 동정과 RNAi를 이용한 기능연구)

  • Lee, Hyun-Seo;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.293-303
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    • 2007
  • Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.

Alteration of Lipid Metabolism Related Proteins in Liver of High-Fat Fed Obese Mice (고지방식이 비만쥐의 지방관련 단백질의 변화)

  • Seo, Eun-Hui;Han, Ying;Park, So-Young;Koh, Hyong-Jong;Lee, Hye-Jeong
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1019-1026
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    • 2010
  • Obesity and being overweight are strongly associated with the development of metabolic disease such as diabetes, hypertension, dyslipidemia. High-fat diet (HFD) is one of the most important factors which cause obesity. In this study, C57BL/6 mice were treated with a HFD for 22 weeks in order to induce obesity and hyperglycemia. Twenty-two weeks later, body weight and plasma glucose level of the HFD group were significantly increased, compared with the normal diet (ND) group. Intra-peritoneal glucose tolerance test (IPGTT) showed glucose intolerance in the HFD group compared with the ND group. These results confirmed that a HFD induced obesity and hyperglycemia in C57BL/6 mice. Plasma levels of triglyceride (TG) and total cholesterol (TC) were increased in the HFD group compared with the ND group. Hepatic levels of TG and TC were also increased by a HFD. To investigate the alteration of lipid metabolism in liver, proteins which are related to lipid metabolism were observed. Among lipid synthesis related enzymes, fatty acid synthase (FAS) and glycerol phosphate acyl transferase (GPAT) were significantly increased in the HFD group. Apolipoprotein B (apoB) and microsomal triglyceride transport protein (MTP), which are related to lipid transport, were significantly increased in the HFD group. Interestingly, protein level and phosphorylation of AMP-activated protein kinase (AMPK), which is known as a metabolic regulator, were significantly increased in the HFD group compared with the ND group. In the present study we suggest that HFD may physiologically increase the proteins which are related with lipid synthesis and lipid transport, but that HFD may paradoxically induce the activation of AMPK.

Chemical Compositions of the Four Lines of Korean Native Chickens (4계통 재래종 닭고기의 화학적 특성)

  • Lee, Kyu Chul;Lee, Sung-Ki;Kim, Hye Kyung
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.119-128
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    • 2016
  • This study was conducted to compare nutritional composition and taste-related compounds of breast and thigh meats from four lines of Korean native chickens (KNC) Yeonsan Ogye, Hyunin Black, Hwangbong, and Hoengseong Yakdak. White Leghorn (WL) was used as a control. Fifteen male chickens (three chickens in each line) were grown under same condition and slaughtered at 53 weeks old. The contents of Fe and K in KNC, especially Hwangbong breast meat and thigh meat of Hyunin Black and Hoengseong Yakdak, were higher than WL. The contents of Na were lower in KNC compared with WL regardless of parts (p<0.05). Vitamin A contents were higher in thigh meat of WL and Hyunin Black, and vitamin $B_1$ contents were lower in Hoengseong Yakdak than other lines. Vitamin $B_3$ were higher in breast meat of four lines of KNC and thigh meat of Hoengseong Yakdak than WL. Total amino acid contents were higher in breast meat of KNC than WL. The level of good-tasting amino acids were significantly higher in breast meat of Hoengseong Yakdak and WL, and thigh meat of Hyunin Black and WL than other lines of chickens (p<0.05). The ratio of good-tasting amino acids to bitter tasting amino acids was higher in breast meat of Hoengseong Yakdak and thigh meat of Hyunin Black than WL. IMP contents were higher in Hyunin Black and WL than other lines of chickens. Based on these results, it can be concluded that four lines of KNC may have superior nutritional quality and taste when compared with WL.

Physicochemical Composition and Fermentation Conditions of Sliced, Dried Radish Kimchi with Flying Fish Roe (날치알을 첨가한 무말랭이 김치의 이화학적 성분 및 발효양상)

  • Jang, Mi-Soon;Park, Hee-Yeon;Nam, Ki-Ho;Kim, Min Jeong
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.566-574
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    • 2014
  • This study was conducted to compare the physicochemical composition and fermentation conditions of sliced, dried radish kimchi with flying fish roe (DFFR). The levels of crude protein, crude lipid, and crude ash in DFFR were higher than those in sliced, dried radish kimchi without flying fish roe (control). DFFR also contained higher levels of Fe and Ca, compared to the control. The inosine monophosphate (IMP) content of DFFR and control was 5.63 and 2.64 mg/100 g, respectively. The polyunsaturated fatty acid and DHA contents in DFFR were approximately 5 and 23 times higher than those in the control, respectively. The major free amino acids contained in these samples were arginine, proline, alanine, leucine, and valine. The number of cells belonging to the Leuconostoc species in DFFR was higher than that in the control. In sensory evaluation studies, DFFR scored the highest in terms of appearance, flavor, taste, and texture.

Quality Investigation of Commercial Northern Sand Lance, Ammodytes Personatus Sauces (시판 까나리액젓의 품질조사)

  • CHO Young Je;IM Yeong Sun;LEE Keun Woo;KIM Geon Bae;CHOI Young Joon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.5
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    • pp.612-617
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    • 1999
  • The quality characteristics of 15 kinds of the commercial northern sand lance, Ammodytes Personatus sauce (CNS) of korean traditional salt-fermented fish sauces were evaluated comparing to the traditional northern sand lance, Ammodytes Personatus sauce (TNS). The ranges of chemical compositions of the CNSs were $66.5\~71.0\%$ moisture, $19.3\~24.6\%$ ash, $4.7\~12.0\%$ crude protein, and the pH and salinity were $5.56\~6.47\%,\;24.0\~32.9\%$, respectively. Total nitrogen, amino nitrogen, total free amino acid and total ATP related compounds (sum of ATP$\~$IMP, HxR, Hx and uric acid) were in the ranges of 0.781$\~$ 1,918 g, 445.9$\~$1037.9 mg, 3,258.9$\~$6:562.6mg in 100 ml CNSs, and 4.766$\~$8.989 $\mu$mol in 1 ml CNSs. The CNSs were higher in content of moisture, TMAO, TMA and pH, but lower in content of crude protein, salinity, total nitrogen, amino nitrogen, total ATP related compounds, absorbance at 453 nm and total free amino acid than the TNSs, Both CNS and TNS samples were rich in free amino acids, such as glutamic acid, lysine, alanine, leucine, valine, aspartic acid and isoleucine in the order.

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The inhibitory Effects of Coenzyme Q10 on Melanogenesis of cultured Human Melanocytes and in vivo Guinea Pig Model (Coenzyme Q10의 멜라닌 생성억제효과)

  • 황재성;박원만;안수미;강병영;이병곤;심영철
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.149-162
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    • 2000
  • Coenzyme Q10 is found in all tissues including skin and it is the well-known coenzyme for mitochondrial enzymes. The electron and proton transfer functions of the quinone ring are of fundamental importance for the oxidative phosphorylation pathway to generate energy in the cells. Coenzyme Q10 has been studied as a potent antioxidant molecule in the skin. It is involved in the skin's response to UVR irradiation. The concentration of this antioxidant in UVR exposed skin is higher than in non-exposed skin. However, recent studies have also shown that coenzyme Q10 is one of the first antioxidants to be depleted when skin is UVR-irradiated. This indicates that coenzyme Q10 is primarily involved in defense mechanisms of the skin. Therefore, we questioned whether coenzyme Q10 shows reulatory effect of melanogenesis. Here we report that coenzyme Q10 inhibits melanin neosynthesis of normal human melanocytes grown in culture, and lightens UVB-induced hyperpigmentation of the guinea pig skin in vivo. We treated human melanocytes with 0.05mM to 0.5mM of coenzyme Q10 for a total of two days. This inhibited melanin neosynthesis of cultured human melanocytes dose-dependently. The inhibitory effect of coenzyme Q10 was as effective as kojic acid or vitamin C on cultured human melanocytes. CoQ10 didn't have direct inhibitory effect on tyrosinase activity in in vitro tyrosine hydroxylase activity To further clarify the effect of coenzyme Q10 on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brownish guinea pigs. The UVB intensity was 500mJ/$\textrm{cm}^2$ and the total energy dose was 1,500 mJ/$\textrm{cm}^2$. The animals were exposed to UVB radiation one times a week for three consecutive weeks. Coenzyme Q10, kojic acid, Arbutin, vitamin C(1% in vehicle) or vehicle alone as a control were then topically applied daily to the hyperpigmented areas twelve times per week far four successive weeks. The lightening effect was evaluated by visual scoring, chromameter and immunohistochemistry. Coenzyme Q10 had lightening effect on the UVB-induced hyperpigmentation without any other side effects, whereas another compounds showed weak lightening efficacies. Therefore, these results suggest that coenzyme Q10 may be useful for solving physiological hyperpigmenting problems for cosmetic purposes.

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Relationship between Reactive Oxygen Species and Adenosine Monophosphate-activated Protein Kinase Signaling in Apoptosis Induction of Human Breast Adenocarcinoma MDA-MB-231 Cells by Ethanol Extract of Citrus unshiu Peel (진피 추출물에 의한 인간유방암 MDA-MB-231 세포의 apoptosis 유도에서 ROS 및 AMPK의 역할)

  • Kim, Min Yeong;HwangBo, Hyun;Ji, Seon Yeong;Hong, Su-Hyun;Choi, Sung Hyun;Kim, Sung Ok;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.4
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    • pp.410-420
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    • 2019
  • Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

Effects of Elastic Band Exercise on Body Composition, Blood lipids and AMPK in the Elderly Women (탄력밴드 운동이 여성노인의 체조성, 혈중지질 및 AMPK에 미치는 영향)

  • Choi, Mi-Ri-Na;Ha, Soo-Min;Kim, Do-Yeon
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.995-1007
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    • 2019
  • The purpose of this study was to investigate the effects of 12-week elastic band exercise on body composition, blood lipids and AMPK in 24 elderly female volunteers aged 65-75 years, and they were divided into the combined exercise group(n=12) and the control group(n=12). The elastic band exercise method was to do exercise 3 times a week for 60 minutes per session, 1-4 weeks for low intensity of OMNI-RES 3-4, 5-8 weeks for medium intensity of OMNI-RES 5-6, 9-12 weeks for OMNI-RES 7-8 of high intensity. In order to compare the differences in the groups before and after the elastic band exercise, two-way repeated measures ANOVA was used to verify the interaction between group and time. The difference in the groups of the measured data was paired t-test, the difference between the groups was paired independent t-test, and analysis of covariance ANCOVA was performed to minimize the inter-group error. The statistical significance level of each item was set to .05. As a result, body fat percentage of exercise group significantly decreased (p<.05), and skeletal muscle volume was significantly increased (p<.01). TC, TG and LDL-C were not significantly different between the exercise and control groups, and HDL-C was significantly decreased in the control group (p<.05). AMPK was significantly decreased in the exercise group (p<.001), but there was no significant difference in the control group. According to the covariance analysis to minimize the error of difference between the pre-exercise groups (p<.05), there was significant difference in AMPK of groups after exercise. These results suggest that the 12-week elastic band exercise has a positive effect on the body composition and AMPK of the elderly women.

A Study on the Whitening Effect of Erigeron annuus (L.) Pers. Ethanol Extract on Melanoma Cell (B16F10) (멜라노마 세포(B16F10)에서의 개망초 추출물을 이용한 미백효과에 관한 연구)

  • Joo, Da-Hye;Yoo, Dan-Hee;Lee, Jin-Young
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.148-157
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    • 2019
  • A 70% ethanol extract of Erigeron annuus (L.) Pers. was investigated for its whitening activity for application as a functional ingredient in cosmetic products. At the E. annuus extract concentration of $100{\mu}g/ml$, the electron-donating ability was found to be 67.83%, the tyrosinase inhibitory effect (related to skin-whitening) was 69%, the elastase inhibitory effect (related to skin-wrinkling) was 69%, and the astringent effect was 80%. The $ABTS^+$ radical-scavenging ability was 87% at the $500{\mu}g/ml$ concentration. In the cell viability test measured on melanoma cells, 96% of the cells treated with $100{\mu}g/ml$ of the extract were viable. According to the western blot results, the protein expression of the microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 was decreased by 60.22%, 47.83%, 54.79%, and 67.88%, respectively, at the extract concentration of $100{\mu}g/ml$. The protein expression of phosphorylated extracellular signal regulated kinase (p-ERK) and phosphorylated cAMP response element-binding protein (p-CREB) was decreased with increasing concentrations of the extract. Reverse transcription-polymerase chain reaction of the extract showed that the mRNA expression of MITF, tyrosinase, TRP-1, and TRP-2 was decreased by 86.51%, 85.22%, 74.26%, and 66.66%, respectively, at $100{\mu}g/ml$ extract concentration. The findings suggest that the 70% ethanol extract from E. annuus (L.) Pers. has potential as a cosmeceutical ingredient with whitening effect.

Inhibitory Effect of Protaetiamycine 9 Derived from Protaetia brevitarsis seulensis Larvae on LPS-mediated Inflammation in RAW264.7 Cells (LPS로 자극한 RAW264.7 대식세포에서 흰점박이꽃무지 유충 유래 Protaetiamycine 9의 항염증 효과)

  • Choi, Ra-Yeong;Seo, Minchul;Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.31 no.11
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    • pp.987-994
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    • 2021
  • Our previous studies have reported that antimicrobial peptides (AMPs) derived from the larvae of white-spotted flower chafer (Protaetia brevitarsis seulensis) exert anti-inflammatory and neuroprotective activities. This study explored the anti-inflammatory effects of protaetiamycine 9 (CVLKKAYFLTNLKLRG-NH2), a novel AMP, derived from P. b. seulensis against lipopolysaccharide (LPS)-mediated inflammatory response in RAW264.7 macrophage cells. Protaetiamycine 9 (25, 50, 75, and 100 ㎍/ml) did not cause cytotoxic effects against RAW264.7 cells. The RAW264.7 cells were pre-treated with various concentrations of protaetiamycine 9 (25-100 ㎍/ml) for 1 hr and then exposed to LPS (100 ng/ml) for 24 hr. Protaetiamycine 9 treatments decreased the LPS-induced secretion of inflammatory mediators, such as nitric oxide (NO), in a dose-dependent manner. Protaetiamycine 9 (25-100 ㎍/ml) effectively downregulated the LPS-induced increase in mRNA and the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. Protaetiamycine 9 also suppressed the production and gene expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, compared to the presence of LPS alone. Furthermore, protaetiamycine 9 inhibited the degradation of inhibitory kappa B alpha (IκB-α) and the phosphorylation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. In conclusion, these results suggest that protaetiamycine 9 exhibits LPS-mediated inflammatory responses by blocking IκB-α degradation and MAPK phosphorylation.