• Title/Summary/Keyword: $P_{tac}$

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Effect of Broussonetia papyrifera L. silage on blood biochemical parameters, growth performance, meat amino acids and fatty acids compositions in beef cattle

  • Tao, Hui;Si, Bingwen;Xu, Wencai;Tu, Yan;Diao, Qiyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.5
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    • pp.732-741
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    • 2020
  • Objective: The study was conducted to investigate the effects of Broussonetia papyrifera L. (B. papyrifera) silage on growth performance, serum biochemical parameters, meat quality, and meat amino acids and fatty acids compositions in beef cattle. Methods: Sixty-four male Angus beef cattle were assigned to 4 groups with 4 pens in each group and 4 beef cattle in each pen, and fed with the total mixed ration supplemented with 0%, 5%, 10%, or 15% B. papyrifera silage for 100 days (control group, 5% group, 10% group and 15% group) separately. Results: Beef cattle had significantly higher final body weight (BW) in 15% group, higher average daily gain (ADG) and dry matter intake (DMI) in 5% group, 10% group and 15% group, and higher feed conversion ratio (FCR) in 10% group and 15% group. Significantly higher blood superoxide dismutase (SOD) concentration was noted in 15% group, higher blood total antioxidant capacity (TAC) in 10% group and 15% group, lower 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) in 15% group. Meat had lower pH in 15% group, higher Commission International DeI'Eclairage (CIE) L in 5% group, 10% group, and 15% group, and lower drip loss in 15% group. Greater concentration of meat polyunsaturated fatty acids (PUFA) was observed in 10% group and 15% group, and docosahexaenoic acid (DHA) in 15% group. Conclusion: Diet with 15% B. papyrifera silage could improve performance and increase final BW, ADG, DMI, and FCR, enhance the antioxidant functions by decreasing blood 8-OHdG and MDA and increasing blood SOD and TAC, improve the meat quality by lowing pH and drip loss and increasing CIE L, increase the meat PUFA and DHA concentration. Polyphenols and flavonoids might be the main components responsible for the antioxidant activity and anti-biohydrogenation in the B. papyrifera silage. And B. papyrifera silage could be used as a new feedstuff in beef cattle nutrition.

Plasma adipocytokines and antioxidants-status in Korean overweight and obese females with dyslipidemia

  • Ha, Ae Wha;Jeong, Su Youn;Kang, Nam E;Kim, Woo Kyoung
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.417-424
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    • 2014
  • BACKGROUD/OBEJECTIVES: It is hypothesized that obese people with dyslipidemia is more likely to have increased oxidative stress and decreased antioxidant status, in comparison with the controls who were obese without dyslipidemia. Thus, the aims of the present study were to determine the dietary intakes, plasma adipokines, and antioxidative systems between obese with dyslipidemia and obese without dyslipidemia were investigated. SUBJECTS/METHODS: Female subjects who were between 20 and 55 years old, and whose BMI was 23 or greater were recruited. Subjects who met the criteria of $BMI{\geq}23$, total cholestero ${\geq}200mg/dL$, LDL cholesterol ${\geq}130mg/dL$, and $TG{\geq}110mg/dL$ were categorized Obese with dyslipidemia. Anthropometric measurements and blood biochemical tests were conducted. The diet survey was conducted by a trained dietitian using two days of 24 hour dietary recall. The lipid peroxidation, the plasma total antioxidant capacity (TAC), the activities of antioxidantive enzymes, and various antioxidantive vitamins levels were determined. RESULTS: Plasma adiponectin and leptin levels were also determined. There were no significant differences for age, Body Mass index (BMI), and body fat (%), waist-size between two groups. Obese with dyslipidemia had significantly high levels of total cholesterol, triglyceride, LDL-cholesterol, the ratio of total cholesterol/HDL-C, and the ratio of HDL-C/LDL-C, respectively. Blood alkaline phosphatase level was statistically different between the two groups (P < 0.05). No statistical significance in dietary intake between two groups was shown. In case of obese with dyslipidemia group, the levels of GSH-Px (P < 0.05) and catalase (P < 0.05) as well as adjusted blood retinol (P < 0.05) and tocopherol level (P < 0.05) were significantly low. However, the plasma concentration of leptin was significantly high (P < 0.05). CONCLUSIONS: Obesity with dyslipidemia was shown to have high arthtrogenic index, depleted antioxidant status, and higher blood leptin levels which suggest higher risks of oxidative stress and cardiovascular diseases.

Expression and Antigenicity of Replicase Protein from Snow Mountain-Like Caliciviruses, Korean Isolates (한국형 사람 Calicivirus Replicase 단백의 발현 및 항원성 평가)

  • Chang, Mi-Yoon;Yang, Jai-Myung;Kim, Kyung-Hee
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.151-160
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    • 1997
  • In view of the potential of replicase protein as a diagnostic reagent for human caliciviruses (HuCVs), we have cloned and over-expressed this gene from the Snow Mountain-like Korean strains in Escherichia coli as a fusion protein with glutathione S-transferase (GST), and described the preliminary antigenic characterization of the recombinant products. Each 470bp fragment corresponding to highly conserved region of RNA-dependent RNA polymerase was generated by RT-PCR from stools of two diarrheal children, cloned in pMOSBlue T-vector, and subcloned between the EcoRI and SalI restriction sites of pGEX-4T-3, a GST gene fusion vector, yielding $pGCV_{pol}$. This construct expressed a Snow Mountain-like HuCV replicase under the control of the IPTG-inducible tac promoter. An extract prepared by sonication of the E. coli cell inclusion bodies bearing $pGCV_{pol}$ products was purified and analyzed by SDS-PAGE. After Coomassie blue staining, it was shown that the recombinant replicase migrated on the gels with an approximate molecular mass of 46.5 kDa, that was subsequently cleaved into a 26 kDa GST fragment and a 20.5 kDa replicase protein upon digestion with thrombin protease. The replicase was recognized on immunoblotting with the sera from symptomatic children with the HuCV-associated diarrhea but not by asymptomatic sera from adults. The results presented the first biological activity of individually expressed HuCV replicase subunit and provided important reagents for diagnosis of HuCV infection.

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Expression and Purification of Human Farnesoid X Receptor-Ligand Binding Domain as Soluble Form Using a Dual Cistronic Expression Vector

  • Kang, Hyun;Ye, Micheal B.;Bahk, Young Yil
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.322-328
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    • 2013
  • In this study, we show the expression and purification of the human recombinant farnesoid X receptor (FXR)- ligand binding domain (LBD) protein in E. coli using a double cistronic vector, pACYCDuet-1, as a soluble form. We describe here the expression and characterization of a biologically active $FXR-LBD_{(248-476)}$. When expressed in the influence of bacterial promoters ($P_{T7}$ and $P_{Tac}$) of the single cistronic expression vectors, the human recombinant $FXR-LBD_{(248-476)}$ was found to be totally insoluble. However, by using a double cistronic expression vector, we were able to obtain the human recombinant $FXR-LBD_{(248-476)}$ in a soluble form. To allow for biological activities, we have subcloned into the pACYCDuet-1 vector, expressed in E. coli cells at some optimized conditions, and purified and characterized the human recombinant active $FXR-LBD_{(248-476)}$ proteins using the fluorescence polarization assay. This suggests that the expression of FXR-LBD in a double cistronic vector improves its solubility and probably assists its correct folding for the biologically active form of the proteins. We suggest that this may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.

Studies on the Separation and Preconcentration of Metal Ions by Chelating Resin containing (Polystyrene-divinylbenzene)-thiazolylazo phenol Derivatives(II) ((Polystyrene-divinylbenzene)-thiazolylazo phenol형 킬레이트 수지에 의한 금속이온의 분리 및 농축에 관한 연구(II))

  • Lim, Jae-Hee;Seol, Kyung-Mi;An, Hye-Sook;Chung, Koo-Chun;Lee, Chang-Heon;Lee, Won
    • Analytical Science and Technology
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    • v.9 no.4
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    • pp.364-372
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    • 1996
  • The sorption and desorption properties of U(VI), Th(IV), Zr(IV), Cu(II), Pb(II), Ni(II), Zn(II), Cd(II) and Mn(II) ions on XAD-16-[2-(2-thiazolylazo)-p-cresol](TAC) chelating resin were studied by elution method for selective separation, concentration and recovery of trace metal ions in sea water. The optimum conditions for the sorption of metal ions were examined with respect to flow rate, pH and concentration of buffer solution. The overall capacities of some metal ions on this chelating resin were 0.41mmol U(VI)/g resin, 0.55mmol Th(IV)/g resin, 0.43mmol Cu(II)/g resin, and 0.32mmol Zr(IV)/g resin, respectively. The elution order of metal ions obtained from breakthrough capacity and overall capacity at pH 5.0 was found as Th(IV)>Cu(II)>U(VI)>Zr(IV)>Pb(II)>Ni(II)>Zn(II)>Cd(II)>Mn(II). Desorption of characteristics for metal ions were investigated with desorption agents such as $HNO_3$, HCl, $HClO_4$, $H_2SO_4$, and $Na_2CO_3$. It was found that most of metal ions except Zr(IV) showed high desorption efficiency with 2M $HNO_3$. But, desorption and recovery of Zr(IV) ion were successfully performed with 1M $H_2SO_4$. The resin was applied for separation and preconcentration of trace amount of U(VI) ion from artificial sea water and the recovery of U(IV) was over 96%.

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Characterization and Expression in Escherichi coli of Streptococcus pneumoniae FtsH

  • Kim, Hee-Soo;Lee, Jae-Jung
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.109-115
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    • 2000
  • FtsH is a membrane-bound, ATP-dependent metalloprotease that is involved in a variety of cellular functions including the regulation of responses to heat and stress shock. Previously, we had cloned and sequenced pneumococcal ftsH gene whose deduced amino acid sequence was very similar to those of several gram-positive bacteria and Escherichia coli, except for the N-terminal domain that was responsible for membrane anchoring. In order to better understand the role of Streptococcus pneumoniae FtsH, we expressed pneumococcal ftsH gene in Escherichia coli. When it was expressed from a strong promoter, $P_{tac}$, a considerable amount of the recombinant FtsH was produced, although the prolonged induction resulted in not only accumulation of breakdown products but also ceasing of the further growth of E. coli host. This indicated that the expression of the exogenous ftsH gene was tightly regulated since the excessive FtsH appeared detrimental to bacterial cells. In Western blotting, the pneumococcal FtsH protein, whether native or recombinant, was reactive to anti-E. coli FtsH serum. The observation that FtsH proteins were well conserved throughout the bacterial kingdom and its expression level was fine-tuned suggests an important role for this protein in the stress adaptation which may be related to infecting process by pneumococci.

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Evaluation of Myocardial Oxygen Consumption with $^{11}C$-Acetate and 3D PET/CT: By Applying Recirculation Correction Method and Modified One-Compartmental Tracer Kinetic Modeling ($^{11}C$-Acetate와 3차원 PET/CT를 이용한 심근의 산소 소모량 평가: 재순환 교정법 및 수정 단일구획 추적자 동적 모델 적용)

  • Chun, In-Kook;Hwang, Kyung-Hoon;Lee, Sang-Yoon;Kim, Jin-Su;Lee, Jae-Sung;Shin, Hee-Won;Lee, Min-Kyung;Yoon, Min-Ki;Choe, Won-Sick
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.4
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    • pp.275-284
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    • 2008
  • Purpose: We intended to evaluate myocardial oxygen consumption ($MVO_2)$ by applying recirculation correction and modified one-compartment model to have a reference range of $MVO_2$ in normal young population and to reveal the effect of recirculation on time-activity curve (TAC). Materials and Methods: In nine normal male volunteers with mean age of $26.3{\pm}4.0$, $MVO_2$ was estimated with 925 MBq (25mCi) of $^{11}C$-Acetate (Neuroscience Research Institute, Gachon University of Medicine and Science, Incheon, Korea) and PET/CT (Biograph 6, Siemens Medical Solution, Germany). Analysis software such as $MATLAB^{(R)}$ v7.1 (Mathworks, Inc., United States), $Excel^{(R)}$ 2007 (Microsoft, United States), and $SPSS^{(R)}$ v12.0 (Apache Software Foundation, United States) were used. Twenty three frames were of $12{\times}10$, $5{\times}60$, $3{\times}120$, $2{\times}300's$ duration, respectively. The modified one-compartmental model and the recirculation correction method were applied. Statistical analysis was performed by using Test of Normality, ANOVA and Post-Hoc (Scheffe's) analysis, and p-value less than 0.05 was considered as significant. Results: The normal reference ranges of $MVO_2$ were presented as $3.18-4.64\;{\times}\;10^{-4}\;ml/g/sec$, $1.91-3.94\;{\times}\;10^{-4}\;ml/g/sec$, $4.31-6.40\;{\times}\;10^{-4}\;ml/g/sec$, $2.84-4.53\;{\times}\;10^{-4}\;ml/g/sec$ and $3.42-5.00\;{\times}\;10^{-4}\;ml/g/sec$ in the septum, the inferior wall, the lateral wall, the anterior wall and the entire wall, respectively. In addition, it was noted that the dual exponentiality of the clearance curve is due to the recirculation effect and that the characteristic of the curve is essentially mono-exponential. Conclusion: $^{11}C$-Acetate is a radiotracer worthwhile to assess $MVO_2$. Re-circulated $^{11}C$ can influence TAC of $^{11}C$ in myocadia and so the recirculation correction must be considered when measuring $MVO_2$.

Frequency and Type of Disputed rpoB Mutations in Mycobacterium tuberculosis Isolates from South Korea

  • Jo, Kyung-Wook;Lee, Soyeon;Kang, Mi Ran;Sung, Heungsup;Kim, Mi-Na;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.80 no.3
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    • pp.270-276
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    • 2017
  • Background: A disputed rpoB mutation is a specific type of rpoB mutation that can cause low-level resistances to rifampin (RIF). Here, we aimed to assess the frequency and types of disputed rpoB mutations in Mycobacterium tuberculosis isolates from South Korea. Methods: Between August 2009 and December 2015, 130 patients exhibited RIF resistance on the MTBDRplus assay at Asan Medical Center. Among these cases, we identified the strains with disputed rpoB mutation by rpoB sequencing analysis, as well as among the M. tuberculosis strains from the International Tuberculosis Research Center (ITRC). Results: Among our cases, disputed rpoB mutations led to RIF resistance in at least 6.9% (9/130) of the strains that also exhibited RIF resistance on the MTBDRplus assay. Moreover, at the ITRC, sequencing of the rpoB gene of 170 strains with the rpoB mutation indicated that 23 strains (13.5%) had the disputed mutations. By combining the findings from the 32 strains from our center and the ITRC, we identified the type of disputed rpoB mutation as follows: CTG511CCG (L511P, n=8), GAC516TAC (D516Y, n=8), CTG533CCG (L533P, n=8), CAC526CTC (H526L, n=4), CAC526AAC (H526N, n=3), and ATG515GTG (M515V, n=1). Conclusion: Disputed rpoB mutations do not seem to be rare among the strains exhibiting RIF resistance in South Korea.

Antitumoral and Antioxidant Potential of Egyptian Propolis Against the PC3 Prostate Cancer Cell Line

  • Salim, Elsayed I;Abd El-Magid, Afaf D;Farara, Khalid M;Maria, Dina SM
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7641-7651
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    • 2015
  • It has been shown previously that nutritional supplements rich in polyphenolic compounds play a significant role in prostate cancer chemoprevention. Propolis is a natural, resinous hive product that has several pharmacological activities including antimicrobial, antioxidant, anti-inflammatory, and antitumoral activities. The aim of this study was to compare the cytotoxic, antioxidant and antitumoral activities of an ethanolic extract of Egyptian propolis (EEP) in vitro with an established chemotherapeutic drug such as doxorubicin (DOX), and the effects of their combination against the PC3 human prostate cancer cell line. Cellular viability and $IC_{50}$ levels with EEP, DOX and their (v/v) combination were detected by sulphorhodamine-B (SRB) assay after incubation of PC3 cells for 72h with different doses (0, 0.01, 0.1, 1, 10 and $100{\mu}g/ml$). Two selected doses of $IC_{50}$ and $IC_{25}$ were applied to cells for 24h for antitumor evaluation assay of treatment compounds. EEP and its (v/v) combination with DOX showed significant antitumor potential besides high antioxidant properties of superoxide dismutase (SOD), total antioxidant capacity (TAC), catalase (CAT), nitric oxide (NO) and reduced glutathione (GSH) levels when compared with the control untreated cells. DNA fragmentation assay and semi quantitative RT-PCR analyses for p53 and Bax genes showed that EEP activated cellular apoptosis and increased the mRNA expression levels more than other treatment. In conclusion, EEP alone or in combination with DOX at both doses used here showed greater antioxidant, antiproliferative and apoptotic effects against the PC3 cell lines as compared to treatment with DOX alone. Therefore, EEP could be considered as a promising candidate for prostate cancer chemotherapy.

Benzo[a]pyrene Cytotoxicity Tolerance in Testicular Sertoli Cells Involves Aryl-hydrocarbon Receptor and Cytochrome P450 1A1 Expression Deficiencies

  • Kim, Jin-Tac;Park, Ji-Eun;Lee, Seung-Jin;Yu, Wook-Joon;Lee, Hye-Jeong;Kim, Jong-Min
    • Development and Reproduction
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    • v.25 no.1
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    • pp.15-24
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    • 2021
  • Benzo[a]pyrene (B[a]P) is a potent carcinogen and is classified as an endocrine-disrupting chemical. In mammalian testes, Sertoli cells support spermatogenesis. Therefore, if these cells are negatively affected by exposure to xenotoxic chemicals, spermatogenesis can be seriously disrupted. In this context, we evaluated whether mouse testicular TM4 Sertoli cells are susceptible to the induction of cytotoxicity-mediated cell death after exposure to B[a] P in vitro. In the present study, while B[a]P and B[a]P-7,8-diol were not able to induce cell death, exposure to BPDE resulted in cell death. BPDE-induced cell death is accompanied by the activation of caspase-3 and caspase-7. Depolarization of the mitochondrial membrane and cytochrome c release from mitochondria were observed in benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated cells. These results indicate that TM4 cells are susceptible to apoptosis in a caspase-dependent manner. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that aryl hydrocarbon receptor (AhR) expression was almost undetectable in TM4 cells and that its expression was not altered after B[a]P treatment. This indicates that TM4 cells are nearly AhR-deficient. In TM4 cells, the CYP1A1 protein and its activity were not present. From these results, it is clear that AhR may be a prerequisite for CYP1A1 expression in TM4 cells. Therefore, TM4 cells can be referred to as CYP1A1-deficient cells. Thus, TM4 Sertoli cells are believed to have a rigid and protective cellular machinery against genotoxic agents. In conclusion, it is suggested that tolerance to B[a]P cytotoxicity is associated with insufficient AhR and CYP1A1 expression in testicular Sertoli cells.