• Title/Summary/Keyword: Protein bodies

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Induction of Apoptotic Cell Death by Methanol Extract of Houttuynia cordata Thunb. in A549 Human Lung Carcinoma Cells (어성초 메탄올 추출물에 의한 A549 인체 폐암세포 사멸유도에 관한 연구)

  • Hong, Su-Hyun;Park, Cheol;Hong, Sang-Hoon;Choi, Byung-Tae;Lee, Yong-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1584-1592
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    • 2006
  • Houttuynia cordata Thunb, well known as 'E-Sung-Cho' in Korea, is traditional medicinal plant generally used in Oriental medicine therapy. We previously reported that the water extract of H. cordata inhibited cell proliferation and induced apoptosis in human breast carcinoma cells. In the present study, we investigated the biochemical mechanisms of anti-proliferative effects by the methanol extract of H. cordata (MEHC) in human lung carcinoma A549 cells. It was found that MEHC could inhibit the cell growth in a dose-dependent manner, which was associated with morphological change and apoptotic cell death as determined by formation of apoptotic bodies, DNA fragmentation and increased populations of apoptotic-sub G1 phase cells. Apoptosis of A549 cells by MEHC was also connected with a down-regulation of anti-apoptotic Bcl-2 and inhibitor of apoptosis proteins (IAPs) expression. MEHC treatment induced the proteolytic activation of caspase-3, caspase-8 and caspase-9, and a concomitant inhibition of poly(ADP-ribose) polymerase (PARP), ${\beta}$-catenin and phospholipase (PLC)-${\gamma}$1 protein expression. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. cordata.

A Divalent Immunotoxin Formed by the Disulfide Bond between Hinge Regions of Fab Domain

  • Choe, Seong Hyeok;Kim, Ji Eun;Lee, Yong Chan;Jang, Yeong Ju;Choe, Mu Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.22 no.12
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    • pp.1361-1365
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    • 2001
  • Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

In Vitro α-Amylase, α-Glucosidase, Pancreatic Lipase, Xanthine Oxidase Inhibiting Activity of Agaricus bisporus Extracts

  • Jung Han Kim;Myoung Jun Jang;Youn Jin Park
    • Mycobiology
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    • v.51 no.1
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    • pp.60-66
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    • 2023
  • In this study, the α-amylase inhibitory activity, α-glucosidase inhibitory activity, pancreatic lipase inhibitory activity, and Xanthine Oxidase inhibitory activity of the fruiting body extracts of 5 varieties of Agaricus bisporus (AB) were confirmed. First, the α-amylase inhibitory activity of AB12, AB13, AB18, AB34, and AB40 methanol extracts was lower than that of acarbose, a positive control, in all concentration ranges. The α-glucosidase inhibitory activity of the AB40, AB13, and AB12 methanol extracts at the extract concentration of 1.0 mg/mL was 80.5%, 81.3%, and 78.5%, respectively, similar to that of acarbose, a positive control. The pancreatic lipase inhibitory activity of the methanol extract of Agaricus bisporus fruiting body was significantly lower than that of the positive control orlistat in the concentration range of 50~1.000 (mg/mL). The Xanthine Oxidase inhibitory activity was 0.5~8.0 mg/mL of each extract, which was significantly lower than that of the positive control allopurinol in the same concentration range. However, the Xanthine Oxidase inhibitory activity of AB13 and AB40 at 8.0 mg/mL was about 70%, which was higher than that of other mushrooms. In conclusion, five kinds of Agaricus bisporus fruiting bodies seem to have inhibitory effects on enzymes such as α-amylase, α-glucosidase, pancreatic lipase, and Xanthine Oxidase that degrade starch and protein. In particular, it has an inhibitory effect and a reduction effect on xanthine oxidase that causes gout, so it is expected that it can be developed and used as a food or health supplement with health functional properties through future research.

Dietary supplementation of Eucommia leaf extract to growing-finishing pigs alters muscle metabolism and improves meat quality

  • Zhenglei Shen;Chuxin Liu;Chuangye Deng;Qiuping Guo;Fengna Li;Qingwu W. Shen
    • Animal Bioscience
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    • v.37 no.4
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    • pp.697-708
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    • 2024
  • Objective: The objective of this study was to investigate the influence of dietary supplementation of Eucommia ulmoides leaf extract (ELE) on muscle metabolism and meat quality of pigs with and without pre-slaughter transportation. Methods: In a 43-day feeding experiment, a total of 160 pigs with an initial body weight 60.00±2.00 kg were randomly assigned into four groups in a completely randomized design with 10 replicates. Pigs in groups A and C were fed a basal diet and pigs in groups B and D were fed a basal diet supplemented with 0.5% ELE. Pigs were slaughtered with (group B and D) or without (group A and C) pre-slaughter transport. Muscle chemical composition, postmortem glycolysis, meat quality and muscle metabolome were analyzed. Results: Dietary ELE supplementation had no effect on the proximate composition of porcine muscle, but increased free phenylalanine, proline, citruline, norvaline, and the total free amino acids in muscle. In addition, dietary ELE increased decanoic acid and eicosapentaenoic acid, but decreased heptadecanoic acid, oleic acid, trans-oleic acid, and monounsaturated fatty acids in muscle. Meat quality measurement demonstrated that ELE improved meat water holding capacity and eliminated the negative effects of pre-slaughter transport on meat cooking yield and tenderness. Dietary ELE reduced muscle glycolytic potential, inhibited glycolysis and muscle pH decline in the postmortem conversion of muscle to meat and increased the activity of citrate synthase in muscle. Metabolomics analysis by liquid chromatographic tandem mass spectrometric showed that ELE enhanced muscle energy level, regulated AMP-activated protein kinase (AMPK) signaling, modulated glycogenolysis/glycolysis, and altered the metabolism of carbohydrate, fatty acids, ketone bodies, amino acids, purine, and pyrimidine. Conclusion: Dietary ELE improved meat quality and alleviated the negative effect of pre-slaughter transport on meat quality by enhancing muscle oxidative metabolism capacity and inhibiting glycolysis in postmortem muscle, which is probably involved its regulation of AMPK.

Anthropometric Measurements of the Upper and Lower Body Balance and Exercise Habit among Female College Students in Some Parts of the Gyeongnam (경남 일부 지역 여자 대학생의 상하체 균형 계측치와 운동 습관)

  • Kim, Young-Sik;Yoon, Im-Sil;Lee, Won-Joon;Nam, Jeong-Su;Yoon, Joong-Soo;Youn, Ryea-Min;Jung, Han-Na;Ko, Jae-Shik;Choi, Hyun-Ju
    • Journal of Life Science
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    • v.20 no.4
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    • pp.535-542
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    • 2010
  • This study was carried out to examine anthropometric parameters and related habits in female college students. Mean age, height, and body weight of the subjects in total (n=212) were 20.7 yrs, 161.8 cm, and 53.5 kg, respectively. Anthropometric parameters were measured by bioelectrical impedance analysis. Mean degree of obesity, body mass index (BMI), and waist-hip ratio (WHR) were 98.4%, $20.6\;kg/m^2$, and 0.80 respectively. Surprisingly, 63.7% of the subjects showed unbalanced weak upper bodies compared to their lower bodies. Therefore, they were divided into two groups: unbalanced weak upper body group (UU, n=135) and balanced upper body groups (BU, n=77). Bodyweight, degree of obesity, BMI, and WHR in the UU group were significantly lower than those in the BU group. Amounts of body protein, body fat, and body mineral in the UU group were also significantly lower than those of the BU group. Mid-arm muscle circumference of the UU group ($18.5{\pm}0.8\;cm$) was significantly lower than those of BU group ($20.4{\pm}1.2\;cm$), although mid-arm circumference of the UU group was not significantly different compared to those of the BU group. On a questionnaire on exercise habits, 1.5% of the UU group and 7.8% of the BU group answered that they exercised regularly, and 55.5% of the UU group and 31.2% of the BU group reported that they never exercised (p<0.01). Even in the subjects who exercised, the duration of the exercise was not sufficient because 38.4% of the UU group and 35.8% of the BU group answered that they exercised for less than 30 min. In conclusion, interventions in exercise habits may be needed for female college students who have unbalanced weak upper bodies.

Utilization of various substrates for the cultivation of oak mushrooms (Lentinula edodes) (다양한 기질에 대한 표고버섯 톱밥재배 가능성)

  • Jeong, Yeun Sug;Jang, Yeongseon;Ryoo, Rhim;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.373-383
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    • 2019
  • We investigated the effect of different media components on the characteristics and productivity of Lentinula edodes cultivated in sawdust. Ten substrates were used for this study. Soybean and cotton seed meal were unsuitable substrates as fungal mycelia did not grow to maturity during the incubation period. Lentinula edodes (NIFoS culture number 2462) was grown on the ten substrates, and soybean hull, mixed medium, cotton seed hull and corn grain showed greater effect on the productivity than wheat bran. However, wheat bran induced higher productivity in NIFoS 2778 than the other substrates. Sawdust medium with soybean meal produced smaller fruiting bodies than the other substrates. In contrast, corn meal media produced larger mushrooms than the other substrates. The external characteristics of the mushrooms varied based on the substrates upon which they were grown. This is not surprising given that the substrates differed with respect to carbohydrate and protein content, e.g., cotton seed and soybean meal contained a higher crude protein and crude ash than the other substrates tested.

Construction of a Recombinant Bacillus velezensis Strain as an Integrated Control Agent Against Plant Diseases and Insect Pests

  • Roh, Jong-Yul;Liu, Qin;Choi, Jae-Young;Wang, Yong;Shim, Hee-Jin;Xu, Hong Guang;Choi, Gyung-Ja;Kim, Jin-Cheol;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1223-1229
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    • 2009
  • To construct a new recombinant strain of Bacillus velezensis that has antifungal and insecticidal activity via the expression of the insecticidal Bacillus thuringiensis crystal protein, a B. thuringiensis expression vector (pHT1K-1Ac) was generated that contained the B. thuringiensis cry1Ac gene under the control of its endogenous promoter in a minimal E. coli-B. thuringiensis shuttle vector (pHT1K). This vector was introduced into a B. velezensis isolate that showed high antifungal activities against several plant diseases, including rice blast (Magnaporthe grisea), rice sheath blight (Rhizotonia solani), tomato gray mold (Botrytis cinerea), tomato late blight (Phytophthora infestans), and wheat leaf rust (Puccinia recondita), by electroporation. The recombinant B. velezensis strain was confirmed by PCR using cry1Ac-specific primers. Additionally, the recombinant strain produced a protein approximately 130 kDa in size and parasporal inclusion bodies similar to B. thuringiensis. The in vivo antifungal activity assay demonstrated that the activity of the recombinant B. velezensis strain was maintained at the same level as that of wild-type B. velezensis. Furthermore, it exhibited high insecticidal activity against a lepidopteran pest, Plutella xylostella, although its activity was lower than that of a recombinant B. thuringiensis strain, whereas wild-type B. velezensis strain did not show any insecticidal activity. These results suggest that this recombinant B. velezensis strain can be used to control harmful insect pests and fungal diseases simultaneously in one crop.

Effects of insect growth regulators(IGRs) on vitellogenesis in insect (곤충의 난황형성에 대한 곤충성장조절제의 작용)

  • Lee, Hee-Kwon;Lee, Jong-Jin;Kim, Moo-Key;Lee, Hoi-Seon
    • The Korean Journal of Pesticide Science
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    • v.5 no.4
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    • pp.11-19
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    • 2001
  • This review discusses the effects and roles of insect hormones and insect growth regulators (IGRs) on vitellogenesis in adult insects. Insect vitellogenesis is regulated by hormones such as juvenile hormone (JH), ecdysteroids, and neurosecretory hormones (ovaryecdysteroidogenic hormone : OEH) released by neurosecretory cells, diet, and other elements(male specific protein of sperm fluid). In the fat bodies, the vitellogenins are synthesized by the stimulation of JH released by corpus allatum (CA) and ecdysteroids produced by follicle cells with the ovary in most insects. Furthermore, vitellogenins are released into the hemolymph, transported to the ovarioles by carrier protein, and incorporated into oocytes for the developing ovary. Of IGRs, juvenile hormone and its mimics such as methoprene and pyriproxifen appear to have pharmacological effects such as membrane lysis, destruction of salivary grand and midgut epithlial cells, fat body cells, and ovarian tissue, and also anti-juvenile hormone such as precocenes I and II appear to have specific cytotoxicity such as inhibition of corpus allatum and oocytes development. These results suggest that IGRs may be useful as agents for integrated pest management.

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Expression and Isolation of Limonoid UDP-glucosyltransferase, a Bitterness-reducing Enzyme, in E.coli (감귤의 고미제거 효소인 limonoid UDP-glucosyltransferase의 대장균 내에서의 발현과 이의 분리)

  • K.Cho, So-Mi;Kim, Young-Mee;Kim, Min-Young;Lee, Do-Seung;Kim, Jae-Hoon;Park, Se-Pill;Riu, Key-Zung;Lee, Dong-Sun
    • Food Science and Preservation
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    • v.18 no.2
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    • pp.208-211
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    • 2011
  • Limonoids are abundant as bitter taste in citrus fruit and other plants. Interestingly. limonoid UDP-glucosyltransferase (LUGT) effectively ameliorates the bitterness from limonoid. The high level of LUGT expression in Escherichia coli can result in the formation of insoluble aggregates known as inclusion bodies. We isolated the soluble LUGT protein when this inclusion body was renaturated with ${\beta}$-cyclidextrin treatment after protein denaturation by urea. Our present results suggest that the isolation of LUGT from inclusion body in cells leads to shed light to characterize the enzyme for food industry purposes.

Antioxidative and Anticancer Activities of Ethanol Extract of Millettia erythrocalyx (Millettia erythrocalyx 에탄올 추출물의 항산화 활성 및 항암 활성에 관한 연구)

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Choi, Sun Mi;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.28 no.1
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    • pp.50-57
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    • 2018
  • Millettia erythrocalyx, a species of plant in the Fabaceae family, is widely distributed in the tropical and subtropical regions of the world, such as the Indies, China, and Thailand. The antiviral activity of flavonoids from M. erythrocalyx has been reported; however, the antioxidative and anticancer activities of M. erythrocalyx remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extract of M. erythrocalyx (EEME) and the molecular mechanism of its anticancer activity in human hepatocellular carcinoma HepG2 cells. EEME exhibited significant antioxidative effects, with a concentration at 50% inhibition ($IC_{50}$) value of $2.74{\mu}g/ml$, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay; moreover, it inhibited cell proliferation in a dose-dependent manner in HepG2 cells. Cell cycle analyses showed that EEME induced HepG2 cell accumulation in the subG1 phase in a dose-dependent manner. EEME also induced apoptosis of HepG2 cells, with increases in apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. Treatment with EEME resulted in increased expression of First apoptosis signal (Fas), a death receptor, and Bcl-2-associated X protein (Bax), a proapoptotic protein, and the activation of caspase-3, 8, and 9, resulting in the cleavage of poly (Adenosine diphosphate-ribose) polymerase (PARP). Collectively, these results suggest that EEME may exert an anticancer effect in HepG2 cells by inducing apoptosis via both the intrinsic and extrinsic pathways.