• 제목/요약/키워드: V-R mechanism

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Role of the Salt Bridge Between Arg176 and Glu126 in the Thermal Stability of the Bacillus amyloliquefaciens ${\alpha}$-Amylase (BAA)

  • Zonouzi, Roseata;Khajeh, Khosro;Monajjemi, Majid;Ghaemi, Naser
    • Journal of Microbiology and Biotechnology
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    • 제23권1호
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    • pp.7-14
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    • 2013
  • In the Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), the loop (residues 176-185; region I) that is the part of the calcium-binding site (CaI, II) has two more amino acid residues than the ${\alpha}$-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region II (residues 118-130), but this interaction is lost in BLA owing to substitution of R176Q and E126V. The goal of the present work was to quantitatively estimate the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of the corresponding salt bridge, Glu126 was deleted (${\Delta}$E126) and converted to Val (E126V), Asp (E126D), and Lys (E126K) by site-directed mutagenesis. Kinetic constants, thermodynamic parameters, and structural changes were examined for the wild-type and mutated forms using UV-visible, atomic absoption, and fluorescence emission spectroscopy. Wild-type exhibited higher $k_{cat}$ and $K_m$ but lower catalytic efficiency than the mutant enzymes. A decreased thermostability and an increased flexibility were also found in all of the mutant enzymes when compared with the wild-type. Additionally, the calcium content of the wild-type was more than ${\Delta}E126$. Thus, it may be suggested that ionic interaction could decrease the mobility of the discussed region, prevent the diffusion of cations, and improve the thermostability of the whole enzyme. Based on these observations, the contribution of loop destabilization may be compensated by the formation of a salt bridge that has been used as an evolutionary mechanism or structural adaptation by the mesophilic enzyme.

High-velocity ballistics of twisted bilayer graphene under stochastic disorder

  • Gupta, K.K.;Mukhopadhyay, T.;Roy, L.;Dey, S.
    • Advances in nano research
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    • 제12권5호
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    • pp.529-547
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    • 2022
  • Graphene is one of the strongest, stiffest, and lightest nanoscale materials known to date, making it a potentially viable and attractive candidate for developing lightweight structural composites to prevent high-velocity ballistic impact, as commonly encountered in defense and space sectors. In-plane twist in bilayer graphene has recently revealed unprecedented electronic properties like superconductivity, which has now started attracting the attention for other multi-physical properties of such twisted structures. For example, the latest studies show that twisting can enhance the strength and stiffness of graphene by many folds, which in turn creates a strong rationale for their prospective exploitation in high-velocity impact. The present article investigates the ballistic performance of twisted bilayer graphene (tBLG) nanostructures. We have employed molecular dynamics (MD) simulations, augmented further by coupling gaussian process-based machine learning, for the nanoscale characterization of various tBLG structures with varying relative rotation angle (RRA). Spherical diamond impactors (with a diameter of 25Å) are enforced with high initial velocity (Vi) in the range of 1 km/s to 6.5 km/s to observe the ballistic performance of tBLG nanostructures. The specific penetration energy (Ep*) of the impacted nanostructures and residual velocity (Vr) of the impactor are considered as the quantities of interest, wherein the effect of stochastic system parameters is computationally captured based on an efficient Gaussian process regression (GPR) based Monte Carlo simulation approach. A data-driven sensitivity analysis is carried out to quantify the relative importance of different critical system parameters. As an integral part of this study, we have deterministically investigated the resonant behaviour of graphene nanostructures, wherein the high-velocity impact is used as the initial actuation mechanism. The comprehensive dynamic investigation of bilayer graphene under the ballistic impact, as presented in this paper including the effect of twisting and random disorder for their prospective exploitation, would lead to the development of improved impact-resistant lightweight materials.

Pharmacokinetic and Pharmacodynamic Characterization of Gliclazide in Healthy Volunteers

  • Kim, Ho-Soon;Yun, Min-Hyuk;Kwon, Kwang-Il
    • Archives of Pharmacal Research
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    • 제26권7호
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    • pp.564-568
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    • 2003
  • Pharmacokinetic and pharmacodynamic properties of gliclazide were studied after an oral administration of gliclazide tablets in healthy volunteers. After an overnight fasting, gliclazide tablet was orally administered to 11 volunteers; Additional 10 volunteers were used as a control group (i.e., no gliclazide administration). Blood samples were collected, and the concentration determined for gliclazide and glucose up to 24 after the administration. Standard pharmacokinetic analysis was carried out for gliclazide. Pharmacodynamic activity of the drug was expressed by increase of glucose concentration ($\Delta$PG), by area under the increase of glucose concentration-time curve ($AUC_{$\Delta$PG}$) or by the difference in increase of glucose concentration ($D_{$\Delta$PG}$) at each time between groups with and without gliclazide administration. Pharmacokinetic analysis revealed that $C_{max}, T_{max}$, CL/F (apparent clearance), V/F (apparent volume of distribution) and half-life of gliclazide were $4.69\pm1.38 mg/L, 3.45\pm1.11 h, 1.26\pm0.35 L/h, 17.78\pm5.27 L, and 9.99\pm2.15 h$, respectively. When compared with the no drug administration group, gliclazide decreased significantly the $AUC_{$\Delta$PG}$ s at 1, 1.5, 2, 2.5, 3 and 4 h (p<0.05). The $\Delta$PGs were positively correlated with $AUC_{gliclazide}$ at 1 and 1.5 h (p<0.05), and the correlation coefficient was maximum at 1 h (r = 0.642) and gradually decreased at 4 h after the administration. The $AUC_{$\Delta$PG}$s were positively correlated with $AUC_{gliclazide}$ at 1, 2, 3 and 4 h (p<0.05), and the maximum correlation coefficient was obtained at 2 h (r=0.642) after the administration. The $D_{$\Delta$PG}$ reached the maximum at 1 h, remained constant from 1 h to 3 h, and decreased afterwards. Therefore, these observations indicated that maximum hypoglycemic effect of gliclazide was reached at approximately at 1.5 h after the administration and the effect decreased, probably because of the homeostasis mechanism, in health volunteers.

Haematological and Immunological Response in Lambs Fed on Raw and Variously Processed Cottonseed Meal

  • Nagalakshmi, D.;Sastry, V.R.B.;Agrawal, D.K.;Katiyar, R.C.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권1호
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    • pp.21-29
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    • 2001
  • An experiment was conducted with twenty crossbred male lambs to assess the effect of cotton (Gossypium) seed meal (CSM) on blood constituents and immunity. Lambs were randomly assigned to a reference diet (30% deoiled peanut meal, DPNM) and four test diets containing 40% of either raw, 45 minutes cooked, 1% $Ca(OH)_2$ and iron (1 free gossy-pol, FG : 0.3 Fe) treated CSM (replacing approximately 50%, reference concentrate mixture). These isonitrogenous and isocaloric concentrate mixtures were fed to meet 80% of protein requirements (NRC, 1985) along with ad lib maize hay for 180 days. Blood was collected at 60, 120 and 180 days post feeding. The lambs were sensitized with Brucella abortus S99 antigen after 140 days and were subjected to ELISA and delayed type hypersensitivity. Blood haemoglobin, erythrocyte count, leucocyte count, total protein, total albumin, total globulin, urea, creatinine concentration and aspartate aminotransferase activity in lambs fed on raw or processed CSM were comparable to the values of reference lambs. The higher (p<0.01) blood glucose levels observed in CSM fed lambs at 60 days of feeding was latter reduced to the levels comparable with those on reference diet at 120 and 180 days of feeding. The alanine amino transferase activity was lower in lambs fed raw and cooked CSM containing diets at 120 and 180 days of feeding. A marginal increase in serum iron and alkaline posphatase activity was observed in iron treated group and raw CSM fed lambs, respectively. The humoral immune response and DTH reactivity was lower (p<0.05) in lambs fed raw CSM (consuming 302.83 mg FG/day). Cooking, $Ca(OH)_2$ and iron treatment of raw CSM showed a positive response in alleviating the suppression of immune response owing to the reduced consumption of FG by 40.19, 17.40% and 26.73%, respectively in these diets. The present study thus indicated that consumption of 40% raw CSM (302.83 mg FG/day) though did not affect majority of the haematological and blood biochemical parameters, but markedly suppressed the immune mechanism of lambs.

Identification of Oligosaccharides in Human Milk Bound onto the Toxin A Carbohydrate Binding Site of Clostridium difficile

  • Nguyen, Thi Thanh Hanh;Kim, Jong Woon;Park, Jun-Seong;Hwang, Kyeong Hwan;Jang, Tae-Su;Kim, Chun-Hyung;Kim, Doman
    • Journal of Microbiology and Biotechnology
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    • 제26권4호
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    • pp.659-665
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    • 2016
  • The oligosaccharides in human milk constitute a major innate immunological mechanism by which breastfed infants gain protection against infectious diarrhea. Clostridium difficile is the most important cause of nosocomial diarrhea, and the C-terminus of toxin A with its carbohydrate binding site, TcdA-f2, demonstrates specific abolishment of cytotoxicity and receptor binding activity upon diethylpyrocarbonate modification of the histidine residues in TcdA. TcdA-f2 was cloned and expressed in E. coli BL21 (DE3). A human milk oligosaccharide (HMO) mixture displayed binding with TcdA-f2 at 38.2 respond units (RU) at the concentration of 20 μg/ml, whereas the eight purified HMOs showed binding with the carbohydrate binding site of TcdA-f2 at 3.3 to 14 RU depending on their structures via a surface plasma resonance biosensor. Among them, Lacto-N-fucopentaose V (LNFPV) and Lacto-N-neohexaose (LNnH) demonstrated tight binding to TcdA-f2 with docking energy of −9.48 kcal/mol and −12.81 kcal/mol, respectively. It displayed numerous hydrogen bonding and hydrophobic interactions with amino acid residues of TcdA-f2.

하수처리장 방류수에 존재하는 항생제 내성인자가 하천에 미치는 영향 (Effect of antibiotic resistant factors in effluent of wastewater treatment plant on stream)

  • 장예진;유용재;설우준;차창준;이옥재;채종찬
    • 미생물학회지
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    • 제53권4호
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    • pp.316-319
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    • 2017
  • 하수처리장 방류수와 하천 중의 항생제 내성인자 분포에 대한 상관성을 분석하기 위해 방류수와 상류 하천수, 하류 하천수를 대상으로 항생제 내성유전자와 전파 관련 유전자를 조사하였다. 3개 지점에서 134~183개의 항생제 내성유전자(ARG) 및 전파 관련 유전자(MGE)가 검출되었으며, 1개의 16S rRNA 유전자에 대한 ARG 및 MGE 유전자의 상대적인 총 합이 0.063~0.422 copy로 분석되었다. ARG와 MGE의 수와 존재량은 방류수에서 가장 높게 검출된 반면, 총 세균 수는 가장 적게 검출됨으로서 하수처리 과정에서 사멸된 세균에 포함된 유전자들이 검출된 것으로 판단된다. 또한 MGE의 존재량 양상이 ARG의 존재량과 상관관계를 보임으로서 항생제 내성균들의 내성기작이 자연내성보다는 획득내성일 가능성을 제시하였다.

Transgenic cucumber expressing the 54-kDa gene of Cucumber fruit mottle mosaic virus is highly resistance and protect non-transgenic scions from soil infection

  • Gal-On, A.;Wolf, D.;Antignus, Y.;Patlis, L.;Ryu, K.H.;Min, B.E.;Pearlsman, M.;Lachman, O.;Gaba, V.;Wang, Y.;Yang. J.;Zelcer, A.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.148.2-149
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    • 2003
  • Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

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다층 지하 구조물로의 고고도 전자기파(HEMP) 커플링 현상에 대한 전자기적 모델링 (Electromagnetic Modeling of High Altitude Electromagnetic Pulse Coupling into Large-Scale Underground Multilayer Structures)

  • 강희도;오일영;김정호;육종관
    • 한국전자파학회논문지
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    • 제23권3호
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    • pp.392-401
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    • 2012
  • 본 논문에서는 다층 지하 구조물로의 고고도 전자기파(high altitude electromagnetic pulse: HEMP)의 커플링 현상을 분석하였다. 이를 위하여, 고고도 전자기파에 대한 모델링을 통하여, 고고도 전자기파의 스펙트럼이 100 MHz 이상의 대역에서 -30 dB 이하로 급격히 감소함을 확인하였다. 또한, 고고도 전자기파의 커플링 영역인 다층 지하 구조물은 지구의 표면과 내부를 구성하는 5층 구조물로 가정하여, 본 구조물을 구성하는 물질의 전파 상수(propagation constant)를 바탕으로 투과 현상을 분석하였다. 그 결과, 50 kV/m의 평면파를 입사시켰을 때, 지상에서 100 m 깊이에 위치한 지하 터널에서 0.1 MHz와 1 MHz의 평면파에 대하여 각각 약 10 kV/m와 5 kV/m의 투과 현상이 발생함을 확인하였다. 투과된 전기장의 효과적인 차폐 효과 확보를 위하여 토양층의 감쇠 상수(attenuation constant)를 이용한 자연 차폐 방법과 금속 격자 구조물의 필터링(filtering) 현상을 이용한 차폐 방법을 통하여 각각 최대 20 dB와 90 dB의 차폐 효과를 얻을 수 있음을 분석하였다.

단기간의 당섭취 증가가 인슐린 감수성에 미치는 영향 (Effect of Acutely Increased Glucose Uptake on Insulin Sensitivity in Rats)

  • 김용운;마인열;이석강
    • Journal of Yeungnam Medical Science
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    • 제14권1호
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    • pp.53-66
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    • 1997
  • 인슐린 저항성이란 인슐린의 생물학적 효과가 감소하는 상태를 말하며 당뇨병의 중요한 병리기전이다. 인슐린 저항성은 매우 다양한 특성을 가지고 있으며 유전적인 요인이나 후천적인 요인에 의하여 발생한다. 이제까지 가장 잘 알려진 인슐린 저항성의 유발인자로는 고혈당과 고인슐린증이 있다. 장기간의 고혈당이나 고인슐린증은 가장 중요한 인슐린의 작용조직인 골격근에서 인슐린 수용체의 결합능을 감소시키고 당섭취에 필요한 제 4형 당수송체의 발현을 억제하는 것으로 알려져 있다. 그러나 이들 인자는 모두 골격근에서 당섭취를 증가시키는 것으로서 인슐린 저항성은 말초조직이 세포내로의 과도한 영양소 섭취를 방지하기 위하여 작동시키는 조절기전일 가능성을 생각할 수 있다. 본 연구에서는 단기간의 당섭취 증가가 더 이상의 당섭취를 억제할 수 있는지를 평가하고 그 기전을 규명하고자 흰쥐에서 2시간 동안 다양한 조건으로 당섭취를 유도하고 1 시간의 휴식기간을 가진 후 인슐린 예민도를 측정하고 골격근을 분리하여 인슐린 수용체와 제 4 형 당수송체를 분석하여 당섭취량과 인슐린 감수성과의 상관관계를 규명하고 인슐린 수용체 결합능과 제 4형 당수송체량을 비교하였다. 실험군은 hyperinsulinemic glucose clamp 전에 시행한 당섭취의 조건으로써 5개 군으로 구분하였다. 제 I 군은 대조군으로 생리식염수만 주입하여 기초상태의 인슐린 농도와 혈당을 유지하였으며, 제 II 군은 somatostatin과 포도당을 주입하여 정상 인슐린농도와 고혈당을 유지하였으며, 제 III 군은 포도당을 주입하여 고인슐린증과 고혈당을 유지하였고, 제 IV 군은 인슐린(100 mU/kg/min)과 포도당을 주입하여 초고인슐린증(${\sim}4000{\mu}U/ml$)과 정상혈당(~100 mg/dl)으로 유지하였으며, 그리고 제 V 군은 인슐린과 포도당을 주입하여 초고인슐린증과 고혈당(~200 mg/dl)을 유지하였다. 체중, 공복시 혈당 및 인슐린 농도는 각 군 간에 유의한 차이가 없었다. Hyperinsulinemic euglycemic clamp 전에 주입한 포도당의 총량(gm/kg)은 제 II 군이 $1.88{\pm}0.151$, 제 III 군이 $2.69{\pm}0.239$, 제 IV 군이 $3.54{\pm}0.198$, 그리고 제 V 군이 $4.32{\pm}0.621$이었다. Hyperinsulinemic euglycemic clamp의 평형상태 당제거율(mg/kg min)은 제 I 군이 $16.9{\pm}1.74$, 제 II 군이 $13.5{\pm}0.53$, 제 III 군이 $11.2{\pm}0.52$, 제 IV 군이 $13.2{\pm}0.92$, 그리고 제 V 군이 $10.4{\pm}0.41$로 전체 군 간에 유의한 차이가 있었으며(p<0.001) 각 군간의 비교에서 제 II, III, IV, V 군의 값이 제 I 군에 비하여 유의하게 낮았다 (p<0.05). 전체 실험동물에서 포도당 주입량과 당제거율 사이에 역상관관계 (r=-0.701, p<0.001)를 보였다. 인슐린 수용체 결합과 Western blot으로 분석한 제 4형 당수송체량은 각 군 간에 유의한 차이가 없었다. 이상의 결과로 미루어 단기간의 당섭취 증가가 더이상의 당섭취를 억제할 수 있으나 인슐린 수용체 결합과 제 4형 당수송체의 숫적 감소없이 발생하였다.

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원전에 사용되는 직류전압제어 대전류원의 개발 (Large-Scale Current Source Development in Nuclear Power Plant)

  • 김종호;최규식
    • 한국항행학회논문지
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    • 제28권3호
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    • pp.348-355
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    • 2024
  • 원전해체 시 대용량 처분시스템을 구동하거나 운영 중 제어봉 구동장치 제어시스템과 같은 대규모 측정전류를 요하는 중요설비를 측정 및 테스트하기 위해서는 측정 매체로서 안정적으로 전류를 공급할 수 있는 전류원이 필요하다. 그러나, 흔히 말하는 전원공급기는 전압원으로서 전압만을 내기 때문에 측정부하에 관계없이 일정한 전류를 공급할 수 있는 전류원 역할을 하지 못한다. 전류원은 전압원과 달리 제품을 만들기가 쉽지 않다. 원전 부품의 전류-전압 변환시험과 같은 건전성을 평가하기 위해서는 정상전류치보다 몇십배 훨씬 큰 수십 mA나 암페어 수준의 전류를 인가하여 테스트를 수행해야 하기 때문에 암페어(ampere) 단위의 전류를 공급할 수 있는 전류원이 필수적이다. 따라서 본 개발품에서는 이러한 필요에 의하여 입력이 전압으로 나타나는 전압원을 이용하여 측정대상에 원하는 전류를 일정하게 공급할 수 있는 전압제어으로서 대용량 전류를 공급할 수 있는 직류전압제어 대전류원을 개발하였다. 개발된 제품의의 신뢰성을 확인하기 위하여 원전에서 사용되는 실제의 데이터값을 적용하여 테스트하였으며, 그 결과를 분석하여 선택회로의 정당성을 입증하였다.