• 생물정신의학
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• 제3권2호
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• pp.170-180
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• 1996

Lowered immune function in the senile dementia patients may be related to the abnormal metabolism of amyloid precursor protein(APP). To investigate the passibility of an abnormal metabolism of APP in lymphocytes and the possible role of APP in the activation of lymphocytes in senile dementia patients, immunohistochemical study of rat spleen and fluorescence activated cell sorter analysis(FACS) of human lymphocytes with the specific antigen far each lymphocyte and double fluorescent marker with antibody to APP were performed. After stimulating lymphocyte with phytohemagglutinin(PHA), APP mRNA and protein were extracted and quantitfied and the influence of ${\beta}$-amyloid protein($A{\beta}$) specific antibody on lymphocyte division was investigated. In spleen, the majority of cells showing $A{\beta}$ immunoreactivity was found in the T-sell dependent zone. FACS indicated that around 90% $CD_4(+)$ T-cells and 60% of $CD_8(+)$ T-sell were immunoreactive to $A{\beta}$ specific antibody(mAb 4G8). Northern blot analysis shows that lymphocyte APP mRNA was gradually increased to reach a maximum at 3 days after activation with lectin mitogen PHA. However, the $A{\beta}$ immunoreactivity an cell surface remained constant during stimulation with PHA, indicating that the release of APP(secreted farm of APP) might be increased. A very large increase in soluble APP secretion was observed in T-lymphocyte upon activation, but only law levels in the resting stale. Immunoblot was carried out an the protein obtained from cell lysate after stimulating lymphocyte by applying PHA to the cultured lymphocyte, and the result was that $A{\beta}$ band of immature farm under 116 KDa marker decreased as the duration of culture was increased after PHA stimulation. The monoclonal $A{\beta}$ specific(4G8) and polyclonal APP antibodies did not inhibit the [$^3H$]-thymidine uptake of mitogen-treated lymphocytes significantly, suggesting that mitogenesis can not be inhibited by specific $A{\beta}$ and polyclonal APP antibody. These results suggest that APP is expressed in T-cell and might be closely associated with the function of T-cells.

• 대한화장품학회지
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• 제32권1호
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• pp.35-44
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• 2006

최근에 고분자, 지질 또는 그 밖의 여러 소재를 이용한 캡슐화 연구는 다양한 기능성 성분의 안정화 및 방출 제어를 목적으로 진행된 바 있다. 본 연구는 주요한 기능성 성분들을 캡슐화할 수 있는 또 하나의 새로운 방법으로써, methylmethacrylate (MMA)와 trimethoxysilylpropylmethacrylate (TMPMA)로 구성된 MMA-TMPMA 공중합체를 제조하였다. Poly(MMA-co-TMPMA)로 제조된 구는 속이 빈 형태의 미립구이며 레티놀, 레티닐팔미테이트, 토코페릴아세테이트, 아스코빌테트라이소팔미테이트와 같은 비타민유도체나 유용성감초추출물과 같은 기능성 성분들을 core 성분으로 캡슐화하는데 매우 유용하였다. 제조된 poly(MMA-co-TMPMA) core-shell 형태의 미립구 평균 입도는 $0.1{\sim}10{\mu}m$, 기능성 성분의 로딩 함량은 $15{\sim}25%$이며 로딩 수율은 90% 이상이었다. Poly(MMA-co-TMPMA) core-shell 형태의 미립구에 함유된 기능성 성분의 안정화는 캡슐 자체에 의한 안정화 외에 미립구 표면에 자외선차단 silane 전구체를 도입함으로써 향상시킬 수 있었다. 기능성 성분에 대한 광안정성은 자외선차단 전구체가 함유된 poly(MMA-co-TMPMA) 미립구의 경우, 자외선차단 전구체가 도입되지 않은 미립구에 비해 25%정도 더 항상됨을 확인할 수 있었다.

• 대한바이러스학회지
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• 제29권3호
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• pp.183-193
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• 1999

Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1620-1627
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• 2009

The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

• 한국잠사곤충학회지
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• 제31권2호
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• pp.72-81
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• 1989

천잠의 알에서 난황 주단백질인 vitellin을 분리·정제하고 가잠 및 상잠의 vitellin과 첨잠 nitellin과의 면역학적 특성을 비교하였으며 천잠의 배자발생에 따른 난내 vitellin의 변동을 조사하였다. 1. 전잠의 체액 vitellogenin은 유충기의 토사말기부터 검출되었다. 2. 점잠의 vitellinm은 가잠 및 상잠의 vitellin과 저기영동상의 이동도는 같았으나 면역학적인 특성에 있어서는 비동질성이었다. 3. 천잠 vitellin은 함량은 전체 가용성 난단백질의 46.0%였으며, 배자발생에 따른 vitellin의 변동은 산난후 8일까지는 같은 수준이었나, 10일 이후부터는 감소되는 경향을 보이다가 5개월된 난과 17개월된 난에서는 상당히 미량만이 검출되었다.

• Food Science and Biotechnology
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• 제15권6호
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• pp.838-843
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• 2006

The chemical processes involved in the formation of green pigment in crushed garlic cloves were investigated based on the principle of pink pigmentation in macerated onions. Intact greening and non-greening garlic cloves were either left untreated or heated at $90^{\circ}C$ for 3 min to inactivate enzyme activities. First, a colorless ether soluble compound referred to as color developer reacted with glycine (among all free amino acids) in garlic to form a second compound insoluble in ether. The latter compound then reacted with formaldehyde to yield the green colored pigment. Alliinase activity was necessary for the production of color developer and for the development of green pigment. In greening garlic that had been heat treated, green pigmentation did not proceed due to the heat-inactivation of alliinase, but the addition of alliinase solution into the garlic homogenates restored the pigmentation. However, this phenomenon was not observed in non-greening garlic with or without heat treatment. Finally, the mechanism of green pigment formation in crushed garlicis similar to that of pink pigment formation in macerated onions.

• 한국수산과학회지
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• 제43권6호
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• pp.606-613
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• 2010

In order to elucidate a mechanism responsible for the development of the odor characteristics of cooked, desirable-flavored shellfish, oysters were extracted using various solvents and the resulting extracts were evaluated organoleptically after cooking. The 80% aqueous methanol extract was found to produce a desirable cooked flavor. This oyster extract was fractionated using ion-exchange column chromatography and dialysis, and each of the fractions was subjected to cooking, followed by organoleptic evaluation. The outer dialysate fraction such as acidic and amphoteric water-soluble fractions produced a cooked oyster flavor. The volatile flavor compounds identified from cooked oyster included 29 hydrocarbons, 20 alcohols, 16 acids, 12 aldehydes, nine nitrogen-containing aromatic compounds, eight ketones, five furans, three esters, three phenols, and one benzene.

• Mass Spectrometry Letters
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• 제11권1호
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• pp.10-14
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• 2020

Riboflavin is a water-soluble vitamin, which serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide. This study aimed to develop a simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the quantification of riboflavin in the Beagle dog plasma. This method utilized simple protein precipitation with acetonitrile and ¹³C₄, ¹⁵N₂-riboflavin was used as an internal standard (IS). For chromatographic separation, a hydrophilic interaction liquid chromatography (HILIC) column was used with gradient elution. The mobile phase consisted of 0.1% (v/v) aqueous formic acid with 10 mM ammonium formate and acetonitrile with 0.1% (v/v) formic acid. Since riboflavin is an endogenous compound, 4% bovine serum albumin in phosphate buffered saline was used as a surrogate matrix to prepare the calibration curve. The quantification limit for riboflavin in the Beagle dog plasma was 5 ng/mL. The method was fully validated for its specificity, sensitivity, accuracy and precision, recovery, and stability according to the US FDA guidance. The developed LC-MS/MS method may be useful for the in vivo pharmacokinetic studies of riboflavin.

• 상하수도학회지
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• 제30권4호
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• pp.409-415
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• 2016

Magnetite particles were synthesized by co-precipitation of water-soluble 밀 스케일-derived precursor by various concentrations of (0.5, 0.67, 1, 2 N) NaOH and (0.6, 0.8, 1.2, 2.4 N) $NH_4OH$. It is theoretically known that as the concentration of the alkaline additive used in iron oxide synthesis increases, the particle size distribution of that iron oxide decreases. This trend was observed in both kind of alkaline additive used, NaOH and $NH_4OH$. In addition, the magnetite synthesized in NaOH showed a relatively smaller particle size distribution than magnetite synthesized in $NH_4OH$. Crystalline phase of the synthesized magnetite were determined by X-ray diffraction spectroscopy(XRD). The particles were then used as an adsorbent for phosphate(P) removal. Phosphorus adsorption was found to be more efficient in NaOH-based synthesized magnetite than the $NH_4OH$-based magnetite.

• 한국미생물·생명공학회지
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• 제42권2호
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• pp.190-193
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• 2014

CMP-Neu5Ac synthetase는 sialyated 된 glycoconjugates의 전구체로 사용되는 CMP-Neu5Ac를 합성하는데 관여하는 주요 효소이다. Escherichia coli K1에서 유래한 CMP-Neu5Ac synthetase 유전자 (neuA)는 평소 E. coli BL21(DE3)에서 비수용성으로 생성되는데, 이를 수용성 단백질로 생산하고자 여러 가지 샤페론 단백질 동시 발현기술을 이용하였다. 이를 위해, GroEL-ES와 DnaK-DnaJ-GrpE를 암호화하는 pG-KJE8 plasmid와 neuA를 동시 형질전환 시켰고 0.01 mM IPTG와 0.005 mg/ml의 L-arabinose로 유도하여 $20^{\circ}C$에서 발현시켰다. 그 결과, E. coli에서의 수용성 CMP-Neu5Ac Synthetase 생산이 현저하게 증가하였다.