• 제목/요약/키워드: DTT

검색결과 162건 처리시간 0.025초

UPLC 분석을 이용한 맥주보리 품종의 호데인 단백질 분석 및 품종 판별 (Discrimination and Hordein Polypeptide Patterns of Malting Barley Varieties Using UPLC)

  • 윤영미;김양길;강천식;박진천;박태일
    • 한국작물학회지
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    • 제66권4호
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    • pp.326-338
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    • 2021
  • 맥주보리 품종의 C-hordeins 분석을 위해 70% EtOH와 55% IPA가 추출용매로 이용되었으며, B-hordeins의 추출은 알코올성 용매에 1% DTT를 추가하여 수행되었다. D-hordeins은 보리 호데인 단백질 중 가장 분자량이 큰 단백질로 약산 혹은 약염기에서 용해가 이루어진다고 알려져 있으며, 본 연구에서는 pH 8.0의 약염기 용매가 추출용매로 이용되었다. 알코올성 용매 종류에 따른 C-hordeins의 구성과 양의 차이는 확인되지 않았으며 1% DTT가 첨가된 알코올성 용매에 B-hordeins과 C-hordeins이 모두 추출됨이 확인되었다. D-hordeins 추출 조건에서는 B-, C-, D-hordeins 이 모두 분석되었지만 B-hordeins의 머무름 시간은 1% DTT가 함유된 알코올 추출과 차이가 있었다. 국내 육성된 맥주보리 26품종들의 C-hordeins 분석을 통해 머무름 시간 16분에서 18분 사이의 피크 유사성을 가지고 7개의 그룹으로 분류할 수 있었다. 또한 맥주보리 육성 품종 중 사천6호와 두산29호, 진광보리와 제주보리, 호품과 다품보리를 제외한 20개 품종들은 마이너 피크들의 분석을 통해 품종의 구분이 가능하였다.

Transcallosal Ipsilateral Motor Pathway from the Unaffected Hemisphere in a Patient with Traumatic Brain Injury

  • Jang, Woo-Hyuk;Lee, Mi-Young;Kwon, Yong-Hyun;Jang, Sung-Ho
    • The Journal of Korean Physical Therapy
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    • 제26권3호
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    • pp.216-219
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    • 2014
  • Purpose: We presented with a patient with traumatic brain injury who showed a transcallosal ipsilateral motor pathway from unaffected motor cortex to affected extremities, as evaluated by diffusion tensor tractography (DTT). Methods: One patient and six age-matched normal subjects were enrolled. A 42-year-old left hemiparetic male who suffered from brain injury by motor vehicle accident 9 years ago showed a leukomalactic lesion in the right corona radiata and parieto-temporal lobe. His left extremities were completely paralyzed initially, but recovered slowly over 2 years. At the time of the evaluation, he was able to grasp and release an object, and to walk with spastic gait pattern. DTT was performed using 1.5 T with a Synergy-L Sensitivity Encoding head coil. DTT was obtained with termination criteria of FA <0.2 and an angle change > $45^{\circ}c$. Results: The motor tracts of the unaffected (left) hemisphere of the patient and control subjects originated from the motor cortex and descended along the known corticospinal tract without any transcallosal tract. By contrast, the tract of the affected (right) hemisphere originated from the left premotor cortex, descended through the left corona radiata, and then crossed the mid-portion of the corpus callosum. The tract then descended through the known corticospinal tract pathway to the right medulla. Conclusion: We conclude that the transcallosal ipsilateral motor pathway from the unaffected hemisphere appeared to contribute to the motor recovery in this patient.

Comparative Study of Enzyme Activity and Stability of Bovine and Human Plasmins in Electrophoretic Reagents, β-mercaptoethanol, DTT, SDS, Triton X-100, and Urea

  • Choi, Nack-Shick;Hahm, Jeung-Ho;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • 제38권2호
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    • pp.177-181
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    • 2005
  • Effects of common electrophoretic reagents, reducing agents ($\beta$-mercaptoethanol [BME] and DTT), denaturants (SDS and urea), and non-ionic detergent (Triton X-100), on the activity and stability of bovine plasmin (b-pln) and human plasmin (h-pln) were compared. In the presence of 0.1% SDS (w/v), all reagents completely inhibited two plns, whereas SDS (1%) and urea (1 M) denatured plns recovered their activities after removal of SDS by treatment of 2.5% Triton X-100 (v/v). However, reducing agents (0.1 M of BME and DTT) treated plns did not restore their activities. Based on a fibrin zymogram gel, five (from b-pln) and four (from h-pln) active fragments were resolved. Two plns exhibited unusual stability in concentrated SDS and Triton X-100 (final 10%) and urea (final 6 M) solutions. Two bands, heavy chain-2 (HC-2) and cleaved heavy chain-2 (CHC-2), of b-pln were completely inhibited in 0.5% SDS or 3 M urea, whereas no significant difference was found in h-pln. Interestingly, 50 kDa (cleaved heavy chain-1, CHC-1) of b-pln and two fragments, 26 kDa (light chain, LC) and 29 kDa (microplasmin, MP), of h-pln were increased by SDS in a concentration dependent manner. We also found that the inhibition of SDS against both plns was reversible.

Production of Embryos by Intracytoplasmic Sperm Injection (ICSI) in Pig

  • Cho, Hwang-Yun;Cho, Seong-Keun;Park, Mi-Ryung;Kim, Jung-Ho;Kim, Jin-Hoi
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.42-42
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    • 2001
  • The objective of this study was to assess the development of porcine follicular oocytes fertilized by ICSI. Cumulus-oocyte-complexes (COCs) were collected by aspiration from follicles of 2-7 mm in diameter from a local slaughterhouse ovaries. Oocytes matured for 40-44 h were centrifuged at 12,000g for 6 min and then injected with sperm prepared by swim-up procedure in the presence or absence of 5 mM dithiothreitol (DTT). Injected oocytes were cultured in NCSU 23 medium during 6 to 8 days. IVF controls were compared to those of resulting embryos. The results obtained were as. follow: 1, The rates of cleavage and development rates into blastocyst by ICSI were not significantly (P<0.05) different between with (53.0% and 19.7%) or without (48.3% and 23.8%) centrifugation, respectively. 2. The cleavage and developmental rates to blastocyst after ICSI with or without 5mM DTT treated-sperm were not significantly (P<0.05) different (60.4% vs 16.4% and 48.5% vs 22.2%, respectively). 3. The cleavage and the developmental rates to blastocyst were not significantly (P<0.05) different between the zygotes obtained by IVF (51.8% vs. 22.4%) and ICSI (51.4% vs. 21.6%). 4. The number of blastomere in blastocyst stages after IVF or ICSI was not significantly different (46.7 $\pm$2.9 and 41.9$\pm$4.6).

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ER Stress-Induced Jpk Expression and the Concomitant Cell Death

  • Kim Hye Sun;Chung Hyunjoo;Kong Kyoung-Ah;Park Sungdo;Kim Myoung Hee
    • 대한의생명과학회지
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    • 제11권2호
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    • pp.135-141
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    • 2005
  • A Jopock (Jpk), a trans-acting factor associating with the position-specific regulatory element of murine Hoxa-7, has shown to have a toxicity to both prokaryotic and eukaryotic cells when overexpressed. Since Jpk protein harbors a transmembrane domain and a putative endoplasmic reticulum (ER)-retention signal at the N-terminus, a subcellular localization of the protein was analyzed after fusing it into the green fluorescent protein (GFP): Both N-term (Jpk-EGFP) and C-term tagged-Jpk (EGFP-Jpk) showed to be localized in the ER when analyzed under the fluorescence microscopy after staining the cells with ER- and MitoTracker. Since ER stress triggers the ER-stress mediated apoptosis to eliminate the damaged cells, we analyzed the expression pattern of Jpk under ER-stress condition. When MCF7 cells were treated with the ER-stress inducer such as DTT and EGTA, the expression of Jpk was upregulated at the transcriptional level like that of Grp78, a molecular chaperone well known to be overexpressed under ER-stress condition. In the presence of high concentration of ER-sterss inducer (10 mM), about 70 (DTT) to $95\%$ (EGTA) of cells died stronly expressing ($10\~12$ fold) Jpk. Whereas at the low concentration ($0.001\~1.0\;mM$) of the inducer, the expression of Jpk was increased about 2.5 (EGTA) to 5 fold (DTT), which is rather similar to those of ER chaperone protein Grp78. These results altogether indicate that the ER-stress upregulated the expression of Jpk and the excess stress induces the ER-stress induced apoptosis and the concomitant expression of Jpk.

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Difference in Injury of the Corticospinal Tract and Spinothalamic Tract in Patients with Putaminal Hemorrhage

  • Jang, Sung Ho;Seo, Jeong Pyo
    • The Journal of Korean Physical Therapy
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    • 제31권6호
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    • pp.358-362
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    • 2019
  • Purpose: We investigated the difference in injury of the corticospinal tract (CST) and the spinothalamic tract (STT) in patients with putaminal hemorrhage, using diffusion tensor tractography (DTT). Methods: Thirty one consecutive patients with PH and 34 control subjects were recruited for this study. DTT scanning was performed at early stage of PH (7-63 days), and the CST and STT were reconstructed using the Functional Magnetic Resonance Imaging of Brain (FMRIB) Software Library program. Injury of the CST and STT was defined in terms of the configuration or abnormal DTT parameters was more than 2 standard deviations lower than that of normal control subjects. Results: Among 31 patients, all 31 patients (100%) had injury of the CTS, whereas 25 patients (80.6%) had injury of the STT: the incidence of CST injury was significantly higher than that of STT (p<0.05). In detail, 20 (64.5%) of 31 patients showed a discontinuation of the CST in the affected hemisphere; in contrast, 14 patients (45.2%) of 31 patients showed a discontinuation of the STT in the affected hemisphere. Regarding the FA value, 6 (19.4%) of 31 patients and 2 (6.4%) of 31 patients were found to have injury in the CST and STT, respectively. In terms of the fiber number, the same injury incidence was observed in 11 patients (35.5%) in both the CST and STT. Conclusion: The greater vulnerability of the CST appears to be ascribed to the anatomical characteristics; the CST is located anteriorly to the center of the putamen compared with the STT.

Haemophilus Influenzae TPx-GRX의 생화학적 특성연구 (Biochemical characterization of Haemophilus Influenzae TPx-GRX)

  • 이동석;김일한
    • 자연과학논문집
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    • 제14권1호
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    • pp.7-24
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    • 2004
  • Thiol peroxidase인 E.coli AhpC의 아미노산 서열을 database를 이용해 분석하여, TPx와 상동성이 있는 새로운 형태의 Thiol peroxidase를 찾아내었다. 그 중 병원성을 갖는 박테리아인 Haemophilus Influenzae에서 존재하는 TPx와 유사하고, GRX와 함께 fusion 되어있는 새로운 형태의 단백질의 유전자를 클로닝하여 E.coli에서 과발현시켜 분리정제 하였다. 정제된 TPx-GRX는 환원제로 thiol 성분을 갖는 MCO system(Fe, DTT, Oxygen)에 의하여 Glutamine Synthetase(GS)의 불활성화를 방어하는 티올 특이적 향산화활성을 갖고, peroxides를 제거하는 peroxidase 활성을 갖는 것을 밝혔다. 이 결과들로부터 TPx-GRX는 새로운 형태의 Thiol perosidase임을 알 수 있었다. 더 나아가서 이 결과들은 TPx-GRX가 병원성 박테리아에서 oxidative stress를 막는 생리적으로 중요한 역할을 할 것이라는 것을 시사한다.

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설프히드릴 변형 화합물질들에 의한 양배추 포스포리파제 D의 시스테인 잔기의 특성 (Characterization of Cysteine Residues in Cabbage Phospholipase D by Sulfhydryl Group Modifying Chemicals)

  • 고은희
    • 대한화학회지
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    • 제50권5호
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    • pp.362-368
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    • 2006
  • 포스포리파아제 D(PLD)의 8개의 시스테인 잔기들의 특성을 파악하기 위해 설프히드릴(SH)기와 반응하는 각종 화학물질들을 동원하였다. 5,5-다이티오비스(2-니트로벤조산) (DTNB)는 시스테인 잔기의 SH기를 적정하기 위해 이용하였으며, 412nm에서의 환원된 DTNB의 값으로부터 자연 상태의 PLD는 1몰 당 4개의 SH기가 있는 것으로 나타났으나, 8 M의 요소 등으로 3차원 구조를 교란 시킨 변성된 PLD는 8개의 SH기가 적정되었다. 이 결과로 시스테인 잔기의 반(4개)은 외부에 노출되어 있고 그 나머지 반은 내부에 가려져 있다고 추정할 수 있다. SH기 변형 시약인 p-클로로머큐리벤조산(PCMB), 요오드아세트산, 요오드아세트아미드, 그리고 N-에칠마레이미드 등은 모두 PLD를 비활성화 시켰다. 이들 중 다이티오스라이톨(DTT)로 처리했을 때 유일하게 PCMB에 의해 비활성화 된 PLD는 가역적으로 그 활성이 회복되었다. 다양한 작용기를 갖는 다이설파이드들을 이용한 노출된 SH기의 주위 환경을 검토한 결과 음전하나 전하를 띄지 않은 다이설파이드들이 양전하를 띈 시스타민 보다 더 효과적으로 PLD를 비활성화 시키는 것으로 나타났다. 그 이외 시스테인 잔기의 산화-환원 전환이 PLD 활성에 미치는 영향을 과산화수소를 이용하여 검토하였다. 과산화수소 산화에 의해 70% 이상 잃은 PLD 활성은 대부분 DTT에 의해 복원되었다. 이들 결과로부터 양배추 PLD의 시스테인 잔기들이 모두 SH기로 존재한다는 것을 반응을 통해 확인 할 수 있었으며, 또한 외부에 노출된 4개의SH기는 PLD 활성 조절에 지대한 영향을 미치고 있는 것으로 나타났다.

Saccharomyces cerevisiae에서 얻은 Purine Nucleoside Phosphorylase의 반응기작과 효소에 대한 Sulfhydryl Reagent의 영향 (Reaction Mechanism of Purine Nucleoside Phosphorylase and Effects of Reactive Agents for SH Group on the Enzyme in Saccharomyces cerevisiae)

  • 최혜선
    • 미생물학회지
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    • 제32권3호
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    • pp.222-231
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    • 1994
  • Saccharomyces cerevisiae에서 얻은 purine nucleoside phosphorylase (PNP)의 반응 기작을 밝히기 위해 반응속도론적 분석이 수행되어졌다. 반응기작에 PNP${\cdot}$phosphate와 PNP${\cdot}$ribose 1-phosphate의 binary complex가 형성되는 것으로 추정되어진다. Initial velocity와 product inhibition study의 결과는 반응이 ordered bi, bi reaction으로 일어난다는 것과 일치하고 있다. 두 개의 기질중 무기인산이 효소에 먼저 붙고, 그 다음에 nucleoside, 그리고 base가 효소를 떠나는 첫 번째 생성물이고 마지막으로 ribose 1-phosphate가 생성되고 효소는 원래의 상태로 돌아간다. 반응속도론적 분석에 이해 제안된 작용기작은 sulfhydryl reagents인 p-chloromercuribenzoate(PCMB) and 5,5'-dithiobisnitrobenzoate (DTNB)에 의한 효소의 불활성화에 대한 기질으 보호작용의 결과와 일치하고 있다. PNP는 ribose 1-phosphate와 phosphate에 의해 보호되지만 nucleoside나 base에 의해서는 아무런 효과과 없다는 사실은 반응 순서가 효소에 무기인산이 먼저 붙는 ordered bi, bi 기작이라는 것을 지지하고 있다. PCMB 나 DTNB에 의해 불활성화된 PNP는 dithiothreitol(DTT)에 의해서는 활성이 완전히 회복되고 2-mercaptoethanol에 의해서는 77%의 활성이 회복된다는 사실은 효소의 불활성화가 가역적이라는 것을 시사하고 있다. PCMB에 의해 불활성화된 효소는 inosine이 변화하는 기질일때 정상효소보다 높은 $K_m$과 낮은 $V_m$ 값을 보여주고 이런 현상은 DTT 처리시 원래의 상태로 돌아온다. DTNB에 의해 불활성화된 효소는 PCMB 처리시와 비슷하게 정상효소보다 높은 $V_m$ 값을 보이지만 $V_m$ 값은 큰 변화가 없다. S. cerevisiae PNP에서 발견되는 높은 무기인산의 농도에서의 하위단위체간의 음성적 협동성이 PCMB나 DTNB를 처리한 PNP에서는 보이지 않았다.

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