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Fragmentation Analysis of rIAPP Monomer, Dimer, and [MrIAPP + MhIAPP]5+ Using Collision-Induced Dissociation with Electrospray Ionization Mass Spectrometry

  • Kim, Jeongmo;Kim, Ho-Tae
    • Mass Spectrometry Letters
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    • 제12권4호
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    • pp.179-185
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    • 2021
  • Collision-induced dissociation (CID) combined with electrospray ionization mass spectrometry (ESI-MS) was used to obtain structural information on rat islet amyloid polypeptide (rIAPP) monomers (M) and dimers (D) observed in the multiply charged state in the MS spectrum. MS/MS analysis indicated that the rIAPP monomers adopt distinct structures depending on the molecular ion charge state. Peptide bond dissociation between L27 and P28 was observed in the MS/MS spectra of rIAPP monomers, regardless of the monomer molecular ion charge state. MS/MS analysis of the dimers indicated that D5+ comprised M2+ and M3+ subunits, and that the peptide bond dissociation process between the L27 and P28 residues of the monomer subunit was also maintained. The observation of (M+ b27)4+ and (M+ y10)3+ fragment ions were deduced to originate from the two different D5+ complex geometries, the N-terminal and C-terminal interaction geometries, respectively. The fragmentation pattern of the [MrIAPP + MhIAPP]5+ MS/MS spectrum showed that the interaction occurred between the two N-terminal regions of MrIAPP and MhIAPP in the heterogeneous dimer (hetero-dimer) D5+ structure.

Analysis of Myosin Heavy Chain Isoforms from Longissimus Thoracis Muscle of Hanwoo Steer by Electrophoresis and LC-MS/MS

  • Kim, Gap-Don
    • 한국축산식품학회지
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    • 제34권5호
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    • pp.656-664
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    • 2014
  • The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscle by liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer (n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide queries were obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43 queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicate identity or extensive homology, p<0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowse score (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirm the expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8 queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

새우난초와 해오라비난초 종자의 기내발아와 유묘생장에 미치는 배지의 영향 (Effect of culture Media on Asymbiotic Seed Germination and Those Seedling Growth of Calanthe discolor and Habenaria radiata)

  • 정미영;정재동;지선옥
    • 식물조직배양학회지
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    • 제25권3호
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    • pp.189-194
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    • 1998
  • 우리나라 야생란 중 원예적 개발가치가 인정되나 대량 증식체계가 확립되지 않아 원예종으로 보급이 미흡한 새우난초 및 해오라비난초를 중심으로 종자의 기내발아와 유묘의 생장촉진에 미치는 배지의 영향에 대해 실험한 결과는 다음과 같다. 새우난초의 발아 및 유묘 생장은 파종전 초음파를 30분간 처리한 후 MS배지에 파종하여 얻은 유묘를 H$_3$P$_4$배지에 이식하여 육묘했을 때 가장 적합하였다. 해오라비난초는 종자를 H$_3$P$_2$배지에 파종하여 형성된 구근을 H$_3$P$_1$ 배지에 이식하여 맹아시킨 후 H$_1$P$_2$배지에 옮겨 배양했을 때 우량종구의 기내생산이 가능하였다.

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구리-오염 토양에서 토마토 식물의 생장과 스트레스-관련 유전자 발현에 미치는 구리-내성 Pseudomonas의 영향 (Effect of Cu-resistant Pseudomonas on growth and expression of stress-related genes of tomato plant under Cu stress)

  • 김민주;송홍규
    • 미생물학회지
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    • 제53권4호
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    • pp.257-264
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    • 2017
  • Pseudomonas veronii MS1과 P. migulae MS2는 여러 가지의 구리-내성 및 식물 생장 촉진 방법을 갖고 있으며 또한 스트레스 에틸렌의 전구체인 1-aminocyclopropane-1-carboxylic acid (ACC)의 ACC deaminase에 의한 가수분해를 통해 식물에서 비생물적 스트레스를 완화시킬 수 있다. 구리 농도 700 mg/kg 토양에서의 4주간 소규모 토마토 재배 실험에서 MS1과 MS2 접종은 비접종 대조군에 비해 토마토 식물의 지상부와 뿌리 길이 및 습윤중량과 건조중량을 모두 유의성 있게 증가시켰다. 접종 토마토 식물은 비생물적 스트레스로부터 식물을 보호할 수 있는 proline및 산화 스트레스 지표인 malondialdehyde도 비접종 대조군보다 적게 함유하였다. 에틸렌 생합성에 관여하는 ACC synthase 유전자, ACS4와 ACS6 그리고 ACC oxidase 유전자, ACO1와 ACO4는 구리 스트레스를 받는 토마토에서 강하게 발현된 반면 MS1과 MS2 접종 토마토에서는 유의성 있게 감소했다. 또한 금속 결합 단백질인 metallothionein 암호화 유전자인 MT2도 위의 유전자들과 유사한 발현 양상을 보였다. 이 모든 결과들은 이 근권세균들이 구리 스트레스 하의 토마토 식물에 구리 내성을 부여하여 낮은 수준의 구리 스트레스와 생장 촉진을 허용하는 것을 가리킨다.

${\apha}$-Casein의 인산화 위치 규명을 위한 티타늄 다이옥사이드($TiO_2$) 방법의 최적화 (Optimization of $TiO_2$ Method to Identify the Phosphorylation Sites of ${\apha}$-Casein)

  • 김혜정;박자혜;백문창
    • 약학회지
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    • 제52권5호
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    • pp.407-411
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    • 2008
  • Phosphorylation plays the most important role in cell signaling mechanism. Various methods to identify the phosphorylation sites of proteins using tandem mass spectrometry (MS/MS) have been reported recently. Furthermore, the enrichment strategy such as Titanium dioxide ($TiO_2$) method should be combined with MS/MS analysis to effectively identify phosphorylation sites. It is necessary to optimize phosphopeptide-enrichment strategy, $TiO_2$ method in this study, due to the low amount of phosphorylated form followed by analyzing them by MS/MS. To evaluate the several conditions to enrich phosphopeptides using $TiO_2$ method, we used ${\apha}$-casein as a standard phosphoprotein and analyzed a representative phosphopeptide (VPQLEIVPNpSAEER) peak of MS spectrum. Batch is better than column method for binding and 300 g/l DHB in loading buffer is better than lower concentration of DHB. 3% TFA and pH 10.5 shows high efficiency of phosphopeptide-enrichment for washing and elution steps, respectively. Finally we identified various efficient conditions of phosphopeptide-enrichment method using $TiO_2$. This optimized method would assist in reliable identifying thousands of phosphorylation sites existed in low abundance from various complex proteins.

Change of the upper airway after mandibular setback surgery in patients with mandibular prognathism and anterior open bite

  • Lee, Kyungjin;Hwang, Soon Jung
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제41권
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    • pp.51.1-51.8
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    • 2019
  • Purpose: It has been reported before that the amount of pharyngeal airway space (PAS) significantly decreases following mandibular setback (MS) surgery in patients with mandibular prognathism (MP). Further, MP patients with an anterior open-bite (AOB) presentation may show a larger decrease in PAS compared with those without AOB. However, studies on postoperative PAS changes in MP patients with AOB remain rare. This study sought to evaluate changes in PAS and hyoid bone positioning following MS surgery in MP patients with and without AOB. Patients and methods: Twenty patients who underwent two jaw surgery involving MS movement were included. Patients were divided into a non-AOB group (n = 10; overbite > 2 mm) and an AOB group (n = 10; overbite < - 4 mm). Three-dimensional changes in PAS and hyoid bone positioning were compared and statistically evaluated pre- and postoperatively using computed tomography (CT). Results: The mean magnitude of MS was 6.0 ± 2.8 mm and 5.6 ± 3.2 mm in the non-AOB group and AOB group, respectively. The oropharyngeal volume and upper hypopharyngeal volume were significantly reduced after surgery in both the groups (p = 0.006 and p = 0.003), while the retroglossal cross-sectional area was significantly reduced only in the AOB group (p = 0.028). Although the AOB group showed a larger decrease in PAS, the difference was not statistically significant between the groups. The position of the hyoid bone showed significant posterior and inferior displacement only in the AOB group, while the vertical displacement of the hyoid bone showed a statistically significant difference between the two groups. Conclusion: PAS was significantly decreased after MS in both the groups, while only the AOB group presented a statistically significant reduction in the retroglossal cross-sectional area. Vertical displacement of the hyoid bone showed a statistically significant difference between the groups, while the PAS change was not. Surgeons should be aware of potential postoperative airway problems that may arise when performing MS surgeries.

Regional Amyloid Burden Differences Evaluated Using Quantitative Cardiac MRI in Patients with Cardiac Amyloidosis

  • Jin Young Kim;Yoo Jin Hong;Kyunghwa Han;Hye-Jeong Lee;Jin Hur;Young Jin Kim;Byoung Wook Choi
    • Korean Journal of Radiology
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    • 제22권6호
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    • pp.880-889
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    • 2021
  • Objective: This study aimed to investigate the regional amyloid burden and myocardial deformation using T1 mapping and strain values in patients with cardiac amyloidosis (CA) according to late gadolinium enhancement (LGE) patterns. Materials and Methods: Forty patients with CA were divided into 2 groups per LGE pattern, and 15 healthy subjects were enrolled. Global and regional native T1 and T2 mapping, extracellular volume (ECV), and cardiac magnetic resonance (CMR)-feature tracking strain values were compared in an intergroup and interregional manner. Results: Of the patients with CA, 32 had diffuse global LGE (group 2), and 8 had focal patchy or no LGE (group 1). Global native T1, T2, and ECV were significantly higher in groups 1 and 2 than in the control group (native T1: 1384.4 ms vs. 1466.8 ms vs. 1230.5 ms; T2: 53.8 ms vs. 54.2 ms vs. 48.9 ms; and ECV: 36.9% vs. 51.4% vs. 26.0%, respectively; all, p < 0.001). Basal ECV (53.7%) was significantly higher than the mid and apical ECVs (50.1% and 50.0%, respectively; p < 0.001) in group 2. Basal and mid peak radial strains (PRSs) and peak circumferential strains (PCSs) were significantly lower than the apical PRS and PCS, respectively (PRS, 15.6% vs. 16.7% vs. 26.9%; and PCS, -9.7% vs. -10.9% vs. -15.0%; all, p < 0.001). Basal ECV and basal strain (2-dimensional PRS) in group 2 showed a significant negative correlation (r = -0.623, p < 0.001). Group 1 showed no regional ECV differences (basal, 37.0%; mid, 35.9%; and apical, 38.3%; p = 0.184). Conclusion: Quantitative T1 mapping parameters such as native T1 and ECV may help diagnose early CA. ECV, in particular, can reflect regional differences in the amyloid deposition in patients with advanced CA, and increased basal ECV is related to decreased basal strain. Therefore, quantitative CMR parameters may help diagnose CA and determine its severity in patients with or without LGE.

토끼 단일 심방근 세포에서 Na-Ca 교환전류의 특성에 관한 연구 (The Properties of Na-Ca Exchange Current in Single Atrial Cells of ,The Rabbit)

  • 염욱;호원경;서경필
    • Journal of Chest Surgery
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    • 제22권4호
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    • pp.548-561
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    • 1989
  • In single atrial cells isolated from the rabbit the properties of inward current of Na-Ca exchange were investigated using the whole cell voltage clamp technique. The current was recorded during repolarization following brief 2 ms depolarizing pulse to +40 mV from a holding potential of * 70 mV. Followings are the results obtained: 1. When stimulated every 30 seconds, the inward currents were activated and reached peak values 6-12 ms after the beginning of depolarizing pulse. The mean current amplitude was 342 pA/cell. 2. The current decayed spontaneously from the peak activation and the time course of the relaxation showed two different phases fast and slow phase. The time constants were 10-18 ms and 60-140 ms, respectively. 3. The recovery of inward current was tested by paired pulse of various intervals. The peak current recovered exponentially with time constant of 140 ms and 1 p M isoprenaline accelerated the recovery process. 4. Relaxation time course was also affected by pulse interval and time constant of the fast phase was reduced almost linearly according to the decrease of pulse interval between 30 sec and 1 sec. 5. The peak activation was increased in magnitude by long prepulse stimulation, 5 p M Bay K, 1 p M isoprenaline or internal and external application of c-AMP. 6. The relaxation time constant of the fast phase was prolonged by 5 p M Bay K or c-AMP, and shortened by isoprenaline. However the time course of the slow relaxation phase was not so much changed. From the above results, it could be concluded that increase of the calcium current by Bay K or c-AMP results in the potentiation and prolongation of intracellular calcium transient, and the facilitation of Ca uptake by SR might be a mechanism of shortening the time constant of current relaxation by short interval stimulation or isoprenaline.

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대사증후군 남성에서의 혈중 Osteoprotegerin의 아디포넥틴과의 상관성과 비만도에 따른 차이 (Association of Plasma Osteoprotegerin with Adiponectin and Difference according to Obesity in Men with Metabolic Syndrome)

  • 나우리;손정민
    • 대한지역사회영양학회지
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    • 제16권6호
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    • pp.762-770
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    • 2011
  • Osteoprotegerin (OPG) plays a core role in bone reformation by antagonizing the effect of receptor activator of nuclear factor ${\kappa}$-B ligand (RANKL), and mediates vascular calcification in cardiovascular disease patients. Thus, we aimed to examine the relationship between serum OPG levels and cardiovascular factors and inflammatory markers in metabolic syndrome patients (MS). This cross-sectional study included 96 men who visited the diet clinic between May and July 2011. Patients were classified into 2 groups based on NCEP-ATP guidelines: normal and with MS (n = 50 and 46, respectively). Physical measurements, biochemical assay were measured. Serum OPG and IL-6, diponectin and hs-CRP were assessed. MS were aged $50.02{\pm}10.85$ years, and normal patients $52.07{\pm}9.56$ years, with no significant differences. Significant differences were not observed in BMI between the 2 groups. Moreover, significant differences were not observed in serum OPG, however, the serum OPG level ($4.41{\pm}1.86pmol/L$) differed significantly between an overweight MS (BMI > 25) and normal patients. OPG was correlated to age (r = 0.410, p = 0.000), HDL-cholesterol (r = 0.209, p = 0.015), and log adiponectin (r = 0.175, p = 0.042). Multiple regression analyses using the enter method showed that age (${\beta}$ = 0.412, p = 0.000) and BMI (${\beta}$ = 0.265, p = 0.000) considerably affected OPG. In conclusion, out study showed that serum OPG levels are correlated with cardiovascular risk factors, such as BMI, HDL-cholesterol and adiponectin in MS and adiponectin, suggesting that serum OPG has potential as a cardiovascular disease indicator and predictor.

혼합치열기 어린이에서 mutans streptococci에 대한 자일리톨 껌의 항균효과에 관한 연구 (THE ANTICARIOGENIC EFFECTS OF XYLITOL ON MUTANS STREPTOCOCCI IN CHILDREN WITH MIXED DENTITION)

  • 박재홍
    • 대한소아치과학회지
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    • 제34권4호
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    • pp.632-638
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    • 2007
  • Xylitol 껌의 mutans streptococci(MS)에 대한 항균효과를 평가하기 위하여 6세${\sim}$11세 사이의 혼합치열기 어린이 62명을 선정하였다. 4개월 동안 1개의 xylitol 껌을 하루 3회 씹는군, 2개의 xylitol 껌을 하루 3회 씹는 군과 xylitol 껌을 씹지 않는 대조군으로 나누어 하루 3번 식후 최소 5분간 껌을 씹게 한 후 1개월 간격으로 자극성타액을 채취하여 선택배지(mitis salivarius kanamicin bacitracin agar)에 배양한 후 MS 군락 수의 감소를 평가하였다. 1회 1개의 xylitol 껌을 씹는 군에서는 MS의 감소를 보이지 않았고, 1회 2개의 xylitol 껌을 씹는 군에서 4개월째 MS의 유의한 감소를 보였다(P < 0.05). 따라서 혼합치열기 어린이의 경우 하루 3회, 총 10g 정도의 xylitol을 4개월 이상 꾸준히 사용하였을 경우 치아우식증에 대한 예방효과가 있을 것으로 사료된다.

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