• Title/Summary/Keyword: Specific activity

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Characterization of the Nanog 5'-flanking Region in Bovine

  • Choi, Don-Ho;Kim, Duk-Jung;Song, Ki-Duk;Park, Hwan-Hee;Ko, Tae Hyun;Pyao, Yuliya;Chung, Ku-Min;Cha, Seok Ho;Sin, Young-Su;Kim, Nam-Hyung;Lee, Woon-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1383-1391
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    • 2016
  • Bovine embryonic stem cells have potential for use in research, such as transgenic cattle generation and the study of developmental gene regulation. The Nanog may play a critical role in maintenance of the undifferentiated state of embryonic stem cells in the bovine, as in murine and human. Nevertheless, efforts to study the bovine Nanog for pluripotency-maintaining factors have been insufficient. In this study, in order to understand the mechanisms of transcriptional regulation of the bovine Nanog, the 5'-flanking region of the Nanog was isolated from ear cells of Hanwoo. Results of transient transfection using a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the -134 to -19 region contained the positive regulatory sequences for the transcription of the bovine Nanog. Results from mutagenesis studies demonstrated that the Sp1-binding site that is located in the proximal promoter region plays an important role in transcriptional activity of the bovine Nanog promoter. The electrophoretic mobility shift assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. In addition, significant inhibition of Nanog promoter activity by the Sp1 mutant was observed in murine embryonic stem cells. Furthermore, chromatin-immunoprecipitation assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. These results suggest that Sp1 is an essential regulatory factor for bovine Nanog transcriptional activity.

Novel Dioxygenases, HIF-α Specific Prolyl-hydroxylase and Asparanginyl-hydroxylase: O2 Switch for Cell Survival

  • Park, Hyun-Sung
    • Toxicological Research
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    • v.24 no.2
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    • pp.101-107
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    • 2008
  • Studies on hypoxia-signaling pathways have revealed novel Fe(II) and $\alpha$-ketoglutarate-dependent dioxygenases that hydroxylate prolyl or asparaginyl residues of a transactivator, Hypoxia-Inducible $Factor-\alpha(HIF-\alpha)$ protein. The recognition of these unprecedented dioxygenases has led to open a new paradigm that the hydroxylation mediates an instant post-translational modification of a protein in response to the changes in cellular concentrations of oxygen, reducing agents, or $\alpha$-ketoglutarate. Activity of $HIF-\alpha$ is repressed by two hydroxylases. One is $HIF-\alpha$ specific prolyl-hydroxylases, referred as prolyl-hydroxylase domain(PHD). The other is $HIF-\alpha$ specific asparaginyl-hydroxylase, referred as factor-inhibiting HIF-1(FIH-1). The facts (i) that many dioxygenases commonly use molecular oxygen and reducing agents during detoxification of xenobiotics, (ii) that detoxification reaction produces radicals and reactive oxygen species, and (iii) that activities of both PHD and FIH-1 are regulated by the changes in the balance between oxygen species and reducing agents, imply the possibility that the activity of $HIF-\alpha$ can be increased during detoxification process. The importance of $HIF-\alpha$ in cancer and ischemic diseases has been emphasized since its target genes mediate various hypoxic responses including angiogenesis, erythropoiesis, glycolysis, pH balance, metastasis, invasion and cell survival. Therefore, activators of PHDs and FIH-1 can be potential anticancer drugs which could reduce the activity of HIF, whereas inhibitors, for preventing ischemic diseases. This review highlights these novel dioxygenases, PHDs and FIH-1 as specific target against not only cancers but also ischemic diseases.

Effects of Environmental Factors on Growth and Nitrogen Fixation Activity of Kummerowia striata (매듭풀의 생육과 질소고정 활성에 미치는 환경요인의 영향)

  • Song, Seung-Dal;Jung-Sook Park;In-Sook Kim
    • The Korean Journal of Ecology
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    • v.18 no.1
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    • pp.43-54
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    • 1995
  • Effects of environmental factors of N, P, pH, moisture, temperature and oxygen on growth and nitrogen fixation activity of kummerowia striate (Thunb.) Schindler seedling, bearing symbiotic root nodules, were quantitatively analyzed during the growing period. The specific nitrogenase activity (ARA) of nodules showed the maximum value of 187 μmol C₂H₄g fr wt-1 h-1 6 weeks after seeds were germinated. The total nitrogenase activities per plant attained as 1.56, 0.85, 0.09 and 4.0, 1.11, 0.04 μmol C₂H₄hr-1, respectively for the treatments of 1, 3 and 5 mM NO₃ ̄and NH₄+ on the 60th day. While the plant grown in N-free media for 20 days after treatments of 5 mM NH₄+for 40 days resulted in 30 mg fr wt of nodule formation and exhibited the relative activities of 152% and 162% for total and specific ARA in comparison with those of control plant grown with N-free for 60 days. Total biomass and ARA was by 70% and 86% lower in N and P deficiency, respectively. The N and P deficient plot showed 70% and 86% decreases of total biomass and ARA in comparison with those of control. The plant grown with N-free for 20 days after pretreatment with N and P free media for 40 days showed the relative values of 77%, 118% and 150%, respectively for nodule biomass, total and specific ARA in comparison with those of control. The treatment with acid or alkali gradients resulted in significant decreases of nodule biomass and ARA. The optimum temperature and pO₂for ARA were 30°C and 40 kPa, respectively. Two peaks of diurnal variation appeared at 11:00 and 23:00 o'clocks by the continuous light condition. The plants with water stress by temporary wilting point rsulted in 95~97% inhibition for nodule respiration, transpiration and specific ARA. Transpiration and ARA ware recovered to 88% and 38% of those of water unstressed plants, respectively, 6 hours after the plants were rewatered from water stressed condition.

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Purification and characterization of TPx from archeabacteria, Halococcus agglomeratus (고염 원시박테리아(Halococcus agglomeratus)에 존재하는 TPx 분리 및 생화학적 특성연구)

  • Choi, Yong-Soo;Cha, Mee-Kyung;Kim, Il-Han
    • The Journal of Natural Sciences
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    • v.14 no.2
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    • pp.67-82
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    • 2004
  • A thiol-specific antioxidant protein (TSA or TPx) was purified from Halophilic archeabacteria Halococcus agglomeratus, by DEAE-Cellulose, Phnyl, sepharose, Sephadex G-75, Sephacryl S-100, Sephacryl S-200, and Q-Wepharose FF. This protein exhibited the preventeive effect against the inactivation of glutamine synthehase (GS) activity was support by a thiol-reducing equicalent such as dithiothreitol. TPx activity was maximal at NaCl concentration above 500mM. The molecular mass of the protein was determinated to be 22-kDa by SDS-PAGE. The TPx purified from Halococcus agglomeratus seems to be similar to other TPx family, except for the salt requirement for the maximal antioxidant activity.

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Comparison of DNase Activities from Infective Larvae vs Adult Worms of Haemonchus contortus (염전위충 감염자충과 성충의 DNase 활성 비교)

  • 곽동미
    • Journal of Veterinary Clinics
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    • v.21 no.3
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    • pp.248-252
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    • 2004
  • DNase activity from infective larvae of the parasitic nematode Haemonchus contortus was characterized and compared to that from whole worm. DNase activity from infective larvae was detected throughout pHs 4-10, but high activity was detected under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but was significantly inhibited at pH 7.0. The activity produced DNA, fragments with mixtures of 3'-hydroxyls (OH) and 3'-phosphates (P) at each pH with predominance of 3'-P. A unique DNase activity at 37 kDa was identified from infective larvae on zymograms. The 37 kDa DNase was detected only at pH 5.0, but not at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 37 kDa infective larval DNase resemble those of classic acidic DNases (e.g., DNase II). In contrast, 34, 36 and 38.5 kDa DNase activities were shown to be specific for whole worm. This result demonstrated that DNases in H contortus are regulated during development.

A study on evaluation of the image with washed-out artifact after applying scatter limitation correction algorithm in PET/CT exam (PET/CT 검사에서 냉소 인공물 발생 시 산란 제한 보정 알고리즘 적용에 따른 영상 평가)

  • Ko, Hyun-Soo;Ryu, Jae-kwang
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.1
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    • pp.55-66
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    • 2018
  • Purpose In PET/CT exam, washed-out artifact could occur due to severe motion of the patient and high specific activity, it results in lowering not only qualitative reading but also quantitative analysis. Scatter limitation correction by GE is an algorism to correct washed-out artifact and recover the images in PET scan. The purpose of this study is to measure the threshold of specific activity which can recovers to original uptake values on the image shown with washed-out artifact from phantom experiment and to compare the quantitative analysis of the clinical patient's data before and after correction. Materials and Methods PET and CT images were acquired in having no misalignment(D0) and in 1, 2, 3, 4 cm distance of misalignment(D1, D2, D3, D4) respectively, with 20 steps of each specific activity from 20 to 20,000 kBq/ml on $^{68}Ge$ cylinder phantom. Also, we measured the distance of misalignment of foley catheter line between CT and PET images, the specific activity which makes washed-out artifact, $SUV_{mean}$ of muscle in artifact slice and $SUV_{max}$ of lesion in artifact slice and $SUV_{max}$ of the other lesion out of artifact slice before and after correction respectively from 34 patients who underwent $^{18}F-FDG$ Fusion Whole Body PET/CT exam. SPSS 21 was used to analyze the difference in the SUV between before and after scatter limitation correction by paired t-test. Results In phantom experiment, $SUV_{mean}$ of $^{68}Ge$ cylinder decreased as specific activity of $^{18}F$ increased. $SUV_{mean}$ more and more decreased as the distance of misalignment between CT and PET more increased. On the other hand, the effect of correction increased as the distance more increased. From phantom experiments, there was no washed-out artifact below 50 kBq/ml and $SUV_{mean}$ was same from origin. On D0 and D1, $SUV_{mean}$ recovered to origin(0.95) below 120 kBq/ml when applying scatter limitation correction. On D2 and D3, $SUV_{mean}$ recovered to origin below 100 kBq/ml. On D4, $SUV_{mean}$ recovered to origin below 80 kBq/ml. From 34 clinical patient's data, the average distance of misalignment was 2.02 cm and the average specific activity which makes washed-out artifact was 490.15 kBq/ml. The average $SUV_{mean}$ of muscles and the average $SUV_{max}$ of lesions in artifact slice before and after the correction show a significant difference according to a paired t-test respectively(t=-13.805, p=0.000)(t=-2.851, p=0.012), but the average $SUV_{max}$ of lesions out of artifact slice show a no significant difference (t=-1.173, p=0.250). Conclusion Scatter limitation correction algorism by GE PET/CT scanner helps to correct washed-out artifact from motion of a patient or high specific activity and to recover the PET images. When we read the image occurred with washed-out artifact by measuring the distance of misalignment between CT and PET image, specific activity after applying scatter limitation algorism, we can analyze the images more accurately without repeating scan.

A STUDY ON SPECIFIC ACTIVITIES OF ENZYMES IN 7, 12-DIMETHYLBENZ(A)ANTHRACENE(DMBA)-INDUCED RAT SUBMAXILLARY GLAND CARCINOGENESIS (백서의 7, 12-Dimethylbenz(a)anthracene 유도 악하선 종양발암과정에서의 효소 특이활성도에 대한 연구)

  • Shim, Hyun-Goo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.12 no.1
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    • pp.27-40
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    • 1990
  • In recent years, tissue antigens and enzymes that will serve as phenotypic markers for malignant cells are becoming increasingly important as diagnostic aids. This study was undertaken to investigate the specific activities of these enzymes in DMBA-induced rat submaxillary gland carcinogenesis. One hundred and twenty Sprague-Dawley rats about 100 gms of body weight were used. In experimental group, DMBA pellet (5mg) was implanted into right submaxillary gland and sham operation was performed into left gland to serve as control. The animals were sacrificed every three weeks up to 15 weeks. Submaxillary glands were excised on both sides and enzyme assays for ${\gamma}-glutamyl$ transpeptidase (GGT), 5'-Nucleotidase, Ornithine decarboxylase(ODC) and Acetyl-Co A carboxylase were carried out biochemically. The obtained results were as follows ; 1. In control group, there was no significant weight change of submaxillary gland, while experimental group, weight was increased remarkably about 7-fold at 15th week since DMBA implantation. 2. In control group, there was no change in specific activities of enzymes during the experimental period. 3. GGT activity was rapidly increased reaching a peak of 1.766${\pm}$0.082units/mg of DNA, 8-fold greater than that of onset. 4. 5'-Nucleotidase activity was increased reaching a peak of $362.1{\pm}53.2{\mu}moles/mg$ of DNA at 9th week. 5. ODC activity was rapidly increased, reaching a peak of 26.2${\pm}$4.8nmoles/mg of DNA at 9th week and quickly returned to that of control at 15th week. 6. Acetyl-Co A carboxylase activity was rapidly increased earlier than other enzymes, reaching a peak of 0.178${\pm}$0.013units/mg of DNA at 6th week and quickly declined to the control level at 15th week.

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Effect of the Extraction Method on the Soybean Embryo Factor 3 Activity (추출 방법에 따른 대두 배인자 3 역가)

  • Lee, Kyung-Hoon;Chung, Dong-Hyo;Kim, Seong-San;Song, Youn-Ho;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.63-66
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    • 1995
  • Soybean nuclear extracts were prepared to detect SEF3(soybean embryo factor 3), which is presumed to be a trans-acting factor for the expression of the soybean ${\beta}-conglycinin\;{\alpha}'$ subunit gene. To increase the specific activity of DNA probe during labeling with $[{\alpha}-^{32}P]$dATP, dATP was added to a final concentration of 1.1 mM during the chase reaction. It results in approximately four-fold increase of specific activity of the DNA probe. Effects of several modifications in preparation of soybean nuclear extracts were examined. It was found that glycerol is effective to stabilize SEF3 during the preparation of nuclear extracts and polyethylenimine could be used to increase the specific activity of SEF3 in nuclear extracts.

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