• Title/Summary/Keyword: dihydrochloride

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In Vitro and Cellular Antioxidant Activity of a Water Extract of Saururus chinensis

  • Kim, Gyo-Nam;Lee, Jung-Sook;Jang, Hae-Dong
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1332-1336
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    • 2008
  • The water extract of Saururus chinensis was investigated for oxygen radical absorbance capacity (ORAC), reducing capacity, metal chelating activity, and intracellular antioxidant activity using HepG2 cell. When 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) was used for the generation of peroxyl radicals in vitro, S. chinensis extract (SC-E) showed the strong and concentration-dependent scavenging activity through donating protons which could be explained by its reducing property. When hydroxyl radicals were generated in vitro through the addition of $Cu^{2+}$ and $H_2O_2$, SC-E demonstrated the antioxidant activity depending on its concentration. In HepG2 cell model, most of intracellular oxidative stress generated by AAPH was efficiently removed by SC-E. However, when $Cu^{2+}$ without $H_2O_2$ was used as an oxidant in the intracellular assay, SC-E partially reduced the oxidative stress caused by $Cu^{2+}$ in cellular antioxidant activity assay system. These results indicate that SC-E could be utilized for the development of functional foods as antioxidant resource in the near future.

Antioxidant and Antihypertension Effects of Enzyme Hydrolysate from Hippocampus abdominalis (식용 빅벨리 해마(Hippocampus abdominalis) 유래 단백질 가수분해물의 항산화와 항고혈압 효능)

  • Je, Jun-Geon;Kim, Hyun-Soo;Lee, Hyo-Geun;Oh, Jae-Young;Wang, Lei;Rho, Sum;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.2
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    • pp.127-133
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    • 2019
  • Seahorses have long been used as ornamental and medicinal products. The sea horse Hippocampus abdominalis has a beautiful color and unique shape and is also used for ornamental purposes and as a traditional medicine in China. This study examined the value of H. abdominalis as a health functional food or food additive. H. abdominalis was hydrolyzed using seven proteases: flavourzyme, neutrase, alcalase, trypsin, kojizyme, pepsin and protamex. The yields of all of the enzyme hydrolysates were higher than that of the aqueous extract. Of the enzymatic hydrolysates, seahorse Protamex hydrolysate (SHP) gave the highest yield and had excellent antioxidant and angiotensin-I converting enzyme inhibitory activities. It protected Vero cells against oxidative by 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH) and antihypertension in Spontaneously Hypertensive Rats. This study attempted to demonstrate H. abdominalis as a health functional food or food additive in the future.

Competitive Enzyme-Linked Immunosorbent Assay for Detection of Gentamicin Residues in Edible Animal products (축산식품 중에 잔류하는 Gentamicin 검사를 위한 ELISA 개발에 관한 연구)

  • Kim, Jae-Myung;Lee, Mun-Han;Lee, Hang;Ryu, Pan-Dong;Cho, Myung-Haing;Park, Jong-Myung
    • Journal of Food Hygiene and Safety
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    • v.9 no.3
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    • pp.123-131
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    • 1994
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

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Antioxidant Activity and Protective Effect of Leaf Extract from Diospyros lotus on Oxidative Stress of Red Blood Cells (고욤 잎 추출물의 항산화 활성 및 적혈구 산화적 손상에 대한 보호 효과)

  • Kim, Hyeon Soo;Kang, Hyun Ju;Jeon, In Hwa;Mok, Ji Ye;Park, Young Kyun;Shin, Jun Ho;Kim, Jang Ho;Jang, Seon Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.5
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    • pp.631-636
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    • 2013
  • This study was to evaluate the antioxidant properties of the leaf extracts of Diospyros lotus (DLE) on the chemical-induced free radical and rat red blood cell (RBC) oxidative damage in vitro. DLE were prepared by extracting with water. DLE showed the high antioxidant activities on the scavenging of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid)-induced radicals. An antioxidant activities of DLE was similar to the reference antioxidant butylated hydroxytoluene (BHT) and (${\pm}$)6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox). Reducing power of $1,000{\mu}g/mL$ DLE also was similar to the vitamin C. In RBC, oxidative hemolysis induced by the aqueous peroxyl radical generator (2,2'-Azobis (2-methylpropionamidine) dihydrochloride (AAPH)) were significantly suppressed by DLE in a dose-dependent manner. Furthermore, DLE prevented the depletion of cytosolic antioxidant glutathione in RBC damaged with AAPH. These results suggest that DLE may has value as natural product with its high quality antioxidant properties against oxidative stress.

Maturational Changes in Binding Capacity of Fowl Sperm to the Epithelium of the Sperm Storage Tubules during Their Passage through the Male Reproductive Tract

  • Ahammad, Muslah U.;Okamoto, S.;Kawamoto, Yasuhiro;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1199-1203
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    • 2011
  • The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

Antihyperlipidemic and Antioxidant Effects of Insamsansa-eum (Renshenshanzha-yin) in Hypercholesterolemic Rats

  • Kim, Young-Suk;Kang, Kyoung-Suk;Jung, Woo-Sang;Moon, Sang-Kwan;Park, Jung-Mi;Ko, Chang-Nam;Cho, Ki-Ho;Bae, Hyung-Sup;Park, Seong-Uk
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.96-104
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    • 2006
  • Insamsansa-eum (Renshenshanzha-yin, ISE) is a new medicine developed to prevent and treat atherosclerotic diseases. To explore antiatherosclerotic effects of ISE, we evaluated the effects of ISE on serum lipids of hypercholesterolemic rats in vivo, as well as its antioxidant activities in vitro. In vitro, ISE showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radical scavenging activities, and inhibited hemolysis induced by 2,2' -azobis-2-amidinopropane dihydrochloride (AAPH) in a dose-dependent manner. In vivo, ISE significantly inhibited increase of total cholesterol (TC), low-density lipoprotein cholesterol (LDL) and triglyceride (TG) values in both high cholesterol diet and Triton WR-1339 models. It also significantly inhibited decrease of high-density lipoprotein cholesterol (HDL) value in the high cholesterol diet model. Collectively, our data suggest that ISE has the potential to control the risk of atherosclerosis development.

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Cholesterol Biosynthesis from Lanosterol: Development of a Novel Assay Method, Characterization, and Solubilization of Rat Hepatic Microsomal Sterol Δ7-Reductase

  • Lee, Joon-No;Paik, Young-Ki
    • BMB Reports
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    • v.30 no.5
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    • pp.370-377
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    • 1997
  • A novel assay method is described for rapid quantitation of reaction rate of sterol ${\Delta}^7$-reductase (${\Delta}^7$-SR) which catalyzes reduction of the ${\Delta}^7$-double bond of sterols. Of six different organ tissues-liver, small intestine, brain, lung, kidney, and testis-. ${\Delta}^7$-SR activity was detected only in liver (2.30 nmol/min/mg protein) and testis (0.11 nmol/min/mg protein). Using a newly developed method which employs diet-induced enzyme proteins and ergosterol as substrate, we assessed both kinetics ($K_m=210\;{\mu}M$, $V_{max}=1.93\;nmol/min/mg$) and inhibition of the rat hepatic ${\Delta}^7$-SR against well-studied cholesterol lowering agents such as triparanol ($IC_{50}=16\;{\mu}M$). 3-$\beta$-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A) ($IC_{50}=5.2\;{\mu}M$), and trans-1.4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944) ($IC_{50}=0.25\;{\mu}M$). Of the three well-known AY-9944-sensitive cholesterogenic enzymes (i.e., ${\Delta}^7$-SR, sterol ${\Delta}^8$-isomerase, and sterol ${\Delta}^14$-reductase). ${\Delta}^7$-SR was found to be the most sensitive enzyme with a noncompetitive inhibition of this compound ($K_i=0.109\;{\mu}M$). Substrate specificity studies of the microsomal ${\Delta}^7$-SR indicate that the relative reaction rate for 7-dehydrocholesterol and ergosterol are 5.6-fold and 1.6-fold higher than that for lathosterol. ${\Delta}^7$-SR activity was also modulated by feeding rats a diet supplemented with 0.5% ergosterol (>2.6-fold) in addition to 5.0% cholestyramine plus 0.1% lovastatin ($\simeq$5.0-fold). Finally, microsomal ${\Delta}^7$-SR was solubilized by 1.5% 3-[3-(cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and enriched on PEG (0~10%) precipitation, which should be suitable for further purification of the enzyme.

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Anti-cancer Effects of Palbohoichoon-tang on Neuroblastoma Cells (신경아세포종에 대한 팔보회춘탕(八寶廻春湯)의 항암 효과)

  • An, Jung-Hwan;Cho, Mun-Young;Woo, Chan;Shin, Yong-Jin;Shin, Sun-Ho
    • The Journal of Internal Korean Medicine
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    • v.35 no.1
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    • pp.79-91
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    • 2014
  • Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

Changes in chemical properties and cytotoxicity of turmeric pigments by microwave treatment (마이크로파처리에 의한 심황색소의 화학안정성 및 세포독성 변화)

  • Song, EiSeul;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.693-698
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    • 2017
  • Turmeric is a yellow food-coloring spice containing curcuminoids, curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BMC), which have several physiological effects. In the present study, the effect of microwave irradiation on the chemical properties, antioxidant activity, and cytotoxicity of turmeric were investigated. Degradation of turmeric pigments was accelerated upon increase in irradiation time or intensity at 405 nm. Residual levels of curcumin, DMC, and BMC after 5 minutes of irradiation at 700 W were 11.3, 34.4, and 71.2%, respectively. Scavenging activities of turmeric pigment against 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH) peroxyl radical and nitrite were enhanced significantly after microwave radiation. However, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity remained unaffected. Cytotoxic activity of turmeric was significantly reduced, and hydrogen peroxide generated from turmeric increased after microwave irradiation. The results obtained indicate that microwave irradiation affects chemical stability and bioactivity of turmeric pigment. Hence, these effects should be considered when processing foods containing turmeric pigments.

Effects of dietary Antrodia cinnamomea fermented product supplementation on antioxidation, anti-inflammation, and lipid metabolism in broiler chickens

  • Lee, M.T.;Lin, W.C.;Lin, L.J.;Wang, S.Y.;Chang, S.C.;Lee, T.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.7
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    • pp.1113-1125
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    • 2020
  • Objective: This study was investigated the effects of dietary supplementation of Antrodia cinnamomea fermented product on modulation of antioxidation, anti-inflammation, and lipid metabolism in broilers. Methods: Functional compounds and in vitro antioxidant capacity were detected in wheat bran (WB) solid-state fermented by Antrodia cinnamomea for 16 days (FAC). In animal experiment, 400 d-old broiler chickens were allotted into 5 groups fed control diet, and control diet replaced with 5% WB, 10% WB, 5% FAC, and 10% FAC respectively. Growth performance, intestinal microflora, serum antioxidant enzymes and fatty acid profiles in pectoral superficial muscle were measured. Results: Pretreatment with hot water extracted fermented product significantly reduced chicken peripheral blood mononuclear cells death induced by lipopolysaccharide and 2,2'-Azobis(2-amidinopropane) dihydrochloride. Birds received 5% and 10% FAC had higher weight gain than WB groups. Cecal coliform and lactic acid bacteria were diminished and increased respectively while diet replaced with FAC. For FAC supplemented groups, superoxide dismutase (SOD) activity increased at 35 days only, with catalase elevated at 21 and 35 day. Regarding serum lipid parameters, 10% FAC replacement significantly reduced triglyceride and low-density lipoprotein level in chickens. For fatty acid composition in pectoral superficial muscle of 35-d-old chickens, 5% and 10% FAC inclusion had birds with significantly lower saturated fatty acids as compared with 10% WB group. Birds on the 5% FAC diet had a higher degree of unsaturation, followed by 10% FAC, control, 5% WB, and 10% WB. Conclusion: In conclusion, desirable intestinal microflora in chickens obtaining FAC may be attributed to the functional metabolites detected in final fermented product. Moreover, antioxidant effects observed in FAC were plausibly exerted in terms of improved antioxidant enzymes activities, increased unsaturated degree of fatty acids in chicken muscle and better weight gain in FAC inclusion groups, indicating that FAC possesses promising favorable mechanisms worthy to be developed.