• Title/Summary/Keyword: H21/32

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Heterologous Expression of Lignin Peroxidase H2 in Escherichia coli: In Vitro Refolding and Activation

  • Lee, Dong-Ho;Kim, Dong-Hyun
    • BMB Reports
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    • v.32 no.5
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    • pp.486-491
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    • 1999
  • An engineered cDNA from Phanerochaete chrysosporium encoding both the mature and propeptide-sequence regions of lignin peroxidase H2 (Lip H2) was overexpressed in Escherichia coli BL21 (DE3) to evaluate its catalytic characteristics and potential application as a pollution scavenger. All expressed proteins were aggregated in an inactive inclusion body, which might be due to inherent disulfide bonds. Active enzyme was obtained by refolding with glutathione-mediated oxidation in refolding solution containing $Ca^{2+}$, heme, and urea. Propeptide-sequence region was not processed as evidenced by N-terminal sequence analysis. Recombinant Lip H2 (rLip H2) had the same physical properties of the native protein but differed in the $K_{cat}$. Catalytic efficiency ($k_{cat}/K_m$) of rLip H2 was slightly higher than that of the native enzyme. In order to express an active protein, fusion systems with thioredoxin or Dsb A, which have disulfide isomerase activity, were used. The fused proteins expressed by the Dsb A fusion vector were aggregated, whereas half of the thioredoxin fusion proteins were recovered as a soluble form but still catalytically inactive. These results suggest that Lip H2 may not be expressed as an active enzyme in Escherichia coli although the activity can be recovered by in vitro refolding.

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(+)-Catechin is a Potent Inhibitor of Intestinal Absorption of Cholesterol in Rats

  • Noh, Sang K.;Koo, Sung I.;Jiang, Yongzhi
    • Preventive Nutrition and Food Science
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    • v.8 no.1
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    • pp.1-6
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    • 2003
  • Catechins exhibit a hypocholesterolemic effect in cholesterol-fed animals. The present study was conducted to examine whether (+)-catechin influences the absorption of cholesterol in rats. Male Sprague-Dawley rats were fed ad libitum an AIN-93G diet containing soybean oil for 5 wk. Rats with lymph cannulae were infused at 3.0 mL/h for 8 h via a duodenal catheter with a lipid emulsion containing radiolabeled cholesterol with or without (+)-catechin. Lymph was collected hourly for 8 h. The enteral infusion of (+)-catechin significantly lowered the lymphatic absorption of $^{14}$ C-cholesterol (21.1 $\pm$ 3.6% dose/B h) compared with controls infused with the lipid emulsion devoid of (+)-catechin (38.2 $\pm$ 1.2% dose/8 h). The intestinal absorption of $\alpha$-tocopherol (24.2$\pm$3.0% dose/8 h) also was significantly decreased by (+)-catechin infusion, relative to controls (32.2$\pm$2.2% dose/8 h). However, the lymphatic outputs of oleic acid and phospholipid were not affected by enteral (+)-catechin infusion. The results indicate that (+)-catechin has a profound inhibitory effect on the intestinal absorption of cholesterol and $\alpha$-tocopherol without affecting the absorption of fat.

Application of Isolated Tyrosinase Inhibitory Compounds from Persimmon Leaves (감나무 잎으로 부터 분리한 tyrosinase 억제물질의 응용)

  • Cho, Young-Je;An, Bong-Jeun;Kim, Jeung-Hoan
    • Journal of Life Science
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    • v.21 no.7
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    • pp.976-984
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    • 2011
  • Total phenolic content was the highest in 60% ethanol extracts at 21.91 mg/g, and inhibitory activity against tyrosinase of 60% ethanol extracts was higher than ethanol extracts of other concentration. The inhibitory compounds against tyrosinase from Persimmon leaves were purified using Sephadex LH-20, MCI-gel CHP-20 column chromatography with gradient elution. Two purified compounds were isolated as a result. The chemical structures of each compound were determined and identified using $^1H$-NMR and $^{13}C$-NMR, FAB-Mass. The compounds were confirmed as (+)-gallocatechin and prodelphinidin B-3. The tyrosinase inhibitory activities of purified (+)-gallocatechin and prodelphinidin B-3 were 29.5 and 40.2%, respectively. The inhibitory activities of (+)-gallocatechin and prodelphinidin B-3 against melanin biosynthesis in melanoma cell were 32.5 and 46.7%. The safety of essence with tyrosinase inhibitory compounds from persimmon leaves was also assessed by various safety profiles. First, changes in pH (4.90~4.95) and viscosity (23,000~26,000 cP) was not detected for 60 days. Essence also showed stability against temperature and light for 60 days. All these findings suggest that extracts from persimmon leaves have a great potential as a cosmetical ingredient with a potent whitening effect.

Effect of Water Management after Fertilizer Application on Fate and Efficiency of Applied Nitrogen (시식 후 물관리 방법이 실소의 동태 및 이용효율에 미치는 영향)

  • 이변우;명을재;최관호
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.2
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    • pp.157-166
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    • 1995
  • The fate and use efficiency of applied nitrogen were evaluated in a pot experiment with different fertilizers and water management practices during 30days after fertilizer application. N-P-K compound fertilizers, 13-10-1l(F-l) for upland Crop use and 15-10-10(F-3) for rice Crop use, and mixed fertilizer, 21-17-17(F-2) for basal dressing in rice were used. Fertilizers corresponding to 1.8g N were mixed thoroughly with the whole volume of sandy loam soil in a pot. The pots were flooded upto 3cm above soil surface for O(0dF), 10(10dF), 20(20dF), and 30(30dF) days after fertilizer application and all the treatments were flooded continuously from 30 days after fertilizer application. During the flooding period water percolation rate was adjusted to 2.5mm/day. Rice seedlings were transplanted 40 days after fertilizer application. The pH of infiltrated water increased with increasing duration of flooding. The pH of F-2 was higher than those of F-1 and F-3 between which there were no differences. The applied nitrogen remained 23% in F-1, 29% in F-2, and 29.1 % in F-3, and 45.0% in 0dF, 26.6% in 10dF, 24.8% in 20dF, and 20.3% in 30dF as inorganic nitrogen at 63 days after fertilizer application. Nitrogen losses by leaching amounted to 51.3%, 32.1% and 48.1% of applied nitrogen in F-1, F-2 and F-3, respectively. Nitrogen leaching losses increased with increasing duration of flood- ing, amounting to 25.7%, 29.8%, 32.7%, and 35.8% in 0dF, 10dF, 20dF and 30dF, respectively. Gaseous loss of applied nitrogen was greatest in F-2, followed by F-1 and F-3. Total loss of nitrogen due to gaseous volatilization and leaching was greatest in F -1, followed by F -2 and F-3, and were greater in the treatments with longer flooding after fertilizer application. Nitrogen recovery by rice shoot until 72 days after transplanting were 23.2%, 24.7% and 27.4% of applied nitrogen in F-1, F-2 and F-3, respectively and 34.1%, 25.5%, 21.1%, and 21.2% in 0dF, 10dF, 20dF and 30dF, respectively.

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Effect of Electrical Preactivation of Recipient Cytoplasm on Nuclear Remodelling in Nuclear Transplant Rabbit Embryos (수핵란의 전 활성화가 토끼 핵이식 수정란의 핵 재구성에 미치는 효과)

  • 전병균;김윤연;정기화;곽대오;이효종;최상용;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.229-238
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    • 1997
  • Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$\times$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$\pm$0.74 and 15.2$\pm$1.32${\mu}{\textrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.

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A study on the identification of type IIa natural diamonds treated by the HPHT method (HPHT(고온고압)에 의해 처리된 type IIa 천연 다이아몬드의 감별에 관한 연구)

  • 김영출;최현민
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.14 no.1
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    • pp.21-26
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    • 2004
  • Results from PL and Raman spectroscopic analyses of HPHT (high-pressure high-temperature) treated type IIa diamonds are presented, and these spectral characteristics are compared with those of untreated diamonds of similar color and type. We identify a number of significant changes by 325 nm He/Cd laser excitation. Several peaks are removed completely, including H4, H3 system in HPHT treated diamond. The N3 system, however, increased in emission. Also we can find the behaviour of the nitrogen-vacancy related center N-V centers at 575 and 637.1 nm, as observed with 514 nm Ar ion laser excitation. When these centers are present, the FWHM (full width at half maximum) of 637.1 nm luminescence intensities offers a potential means of separating HPHT-treated from untreated type IIa diamonds. The width of 637.1 nm $(N-V)^-$line measured at the position oi half the peak's height are determine to range from 19.8 to $32.1cm^{-1}$ for HPHT treated diamonds.

Effects of Breeder Age and Stocking Density on Performance, Carcass Characteristics and Some Stress Parameters of Broilers

  • Onbasilar, E.E.;Poyraz, O.;Cetin, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.262-269
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    • 2008
  • The aim of this study was to determine the effects of breeder age and stocking density on performance, carcass characteristics and some stress parameters (H-L ratio, serum glucose, cholesterol and triglyceride levels, tonic immobility test (TI), antibody production, relative asymmetry (RA) and external appearances). This experiment was carried out with 705 one-day old male broiler chicks (Ross 308) obtained from three different ages of broiler breeder (32, 48 and 61 wks). Each age group was randomly divided into two stocking density groups (11.9 and 17.5 broilers per $m^2$) with 5 replications per group. The experimental period was 6 weeks. Broilers from 32 wk-old breeders had lower initial weight (p<0.001), body weight gain of the first 3 week of rearing (p<0.01), the percentage of abdominal fat (p<0.001) and serum cholesterol level (p<0.01); higher percentage of gizzard (p<0.01) and longer TI duration (p<0.001) than those from 48 and 61 wk-old breeders. Broilers reared at 17.5 b/m2 had lower final BW, body weight gain, feed consumption, feather condition and foot health (p<0.001), higher percentage of heart, H-L ratio, serum glucose and cholesterol levels (p<0.001), and longer TI durations (p<0.001). There were no significant interactions in examined parameters except for feed to gain ratio between breeder age and stocking density.

Chemical Modification of Rupestonic Acid and Preliminarily In Vitro Antiviral Activity Against Influenza A3 and B Viruses

  • Yong, Jian-Ping;Aisa, Haji Akber
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1293-1297
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    • 2011
  • To improve the biological activities of rupestonic acid, 21 new rupestonic acid fatty ester derivatives (2a-2h) and aromatic ester derivatives (2i-2u) were synthesized and preliminarily evaluated for their anti-influenza activity in vitro by the national center for drug screening of China, using the Oseltamivir and Ribavirin as reference drugs. The results showed that 2l ($IC_{50}=0.5{\mu}mol/L$) exhibited potent anti-influenza $A_3$ viral activity among the synthesized compounds and was 10-fold more potent than that of the reference drug Oseltamivir ($IC_{50}=5.1{\mu}mol/L$).

Comparison of Cold Hardiness in Canes and Buds of Kiwifruit Cultivars (품종에 따른 키위나무 눈과 가지의 내한성 비교)

  • Kim, H.L.;Chae, W.B.;Kim, J.G.;Lee, M.H.;Rhee, H.C.;Kim, S.H.;Kwack, Y.B.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.21 no.1
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    • pp.29-40
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    • 2019
  • In Korea kiwifruit growing area is limited to southern coastal region and Jeju island, partly due to the lack of information on their cold hardiness in winter. This study was carried out to investigate cold hardiness of Korean kiwifruit cultivars in a period of dormancy for using it as preliminary data to expand the cultivation area of kiwifruit in Korea. A total of five kiwifruit cultivars in two species and hybrid, Actinidia deliciosa ('Hayward' and 'Garmrok'), A. chinensis ('Goldone') and A. arguta hybrid ('Bangwoori' and 'Skinny Green') were subjected to five freezing treatments of -12℃, -15℃, -18℃, -21℃ and -24℃. Cell membrane damage in all cultivars initiated in -18℃/32h and cell membrane stability was lost in -24℃ in most cultivars, except for 'Skinny Green'. Cold hardiness was estimated by 50% lethal temperature (LT50) which was determined by triphenyl tetrazolium chloride (TTC) reduction. In branches, LT50 was -15℃ in 'Hayward' and 'Garmrok', -18℃ in 'Bangwoori' and -21℃ in 'Goldone.' The LT50 of buds on 'Hayward' and 'Garmrok' was 56 and 42 hours in -15℃ and 4 and 11 hours in -18℃, respectively; however, LT50 of buds on 'Goldone' was 51 hours in -18℃ and that on 'Bangwoori' was 3 hours in -24℃. Cold hardiness results imply that it may be difficult for cultivars in A. deliciosa such as 'Hayward' and 'Garmrok' to be grown in the north of southern coastal region in Korea; however, it can be possible for several cultivars in A. chinensis and A. arguta hybrid to be grown in the northern part of Korean kiwifruit belt if cold tolerance in the thaw is confirmed.

Chemical Composition in Relation to Quality Evaluation of Korean Honey (한국산꿀의 품질특성)

  • Chung, Won-Chul;Kim, Man-Wook;Song, Ki-Joon;Choi, Eon-Ho
    • Korean Journal of Food Science and Technology
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    • v.16 no.1
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    • pp.17-22
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    • 1984
  • Quality of five kinds of honey were evaluated. Honey had total soluble sugar above 96% on the dry weight and a little amount of protein. The mineral composition varied with the kinds of honey, but all tested honey showed the highest content of K and Na, and a considerable amount of Fe, Mg, Ca, Zn, and P. Free sugars were mainly composed of fructose and glucose. The fructose/glucose ratio of honey was 1.0-1.39. Buckwheat honey showed comparatively higher content of sucrose and maltose. All tested honey were analyzed to give pH 3.23-4.32, total acid 10.5-23.9 meq/kg, hydroxymethylfurfural 0.58-21.31 mg/kg, diastatic activity 13.95-36.59.

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