• 한국식품영양과학회지
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• 제35권5호
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• pp.594-599
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• 2006

이전의 연구에서, 우육중의 주된 알레르겐은 우육추출물 중의 BSA, BGG라고 보고하였다. 본 연구에서는 이들 단백질의 열처리와 초고압처리와 같은 물리적 처리에 따른 SDS-PAGE상의 변화와 우육알레르기 환자의 혈청을 이용한 알레르겐성의 변화를 Western blots로 관찰하였다. 우육추출물에 대한 열처리에 의한 알레르겐 단백질의 변화에서, $100^{\circ}C$ 가열처리군에서 BSA band는 큰 감소를 보였으나, BGG band는 완전히 제거되지 못하고 남아 있었고, 항원성 역시 대조군에 비하여 감소는 하였으나 여전히 유지되고 있었다. $120^{\circ}C$이상의 열처리는 추출물중의 BSA와 BGG band 의 소실과 이에 따른 항원성의 큰 감소를 가져왔다. 우육추출물중의 BSA단백질은 열처리에서와 달리 초고압처리에 의하여 전기영동상에 거의 변화를 보이지 않았다. 그러나 BGG 단백질은 400 MPa 가압처리군부터 전기 영동상 band가 감소하였고, 항원성 역시 유의적으로 감소하였다. 이는 초고압처리 정도에 따른 우육추출물중의 BSA와 BGG를 포함한 여러 분자의 SDS-PAGE상의 특이적 변화가 최초로 시사된 결과이다. 이러한 BGG 단백질의 고압처리에 의한 변화는 추출물중의 어떤 효소에 의한 것으로 추측되었고, 고압처리 후 $37^{\circ}C$숙성 실험으로 이러한 가설이 확인되었다. 우육추출물에 존재하고 초고압처리로 활성화되어 BGG분자에 특이적으로 관여하는 효소 및 이와 관련된 메커니즘에 대한 연구가 앞으로 필요할 것으로 보인다.

• 대한핵의학회지
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• 제14권1호
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• pp.45-56
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• 1980

$T_3-BSA\;and\;T_4-BSA$ conjugates were prepared and identified spectrophotometrically. The ${\lambda}max$ of the conjugates was just coincided with that of BSA, but the molar extinction coefficients of the conjugates were generally larger than that of BSA itself. The molar ratios of $T_3:\;BSA\;and\;T_4:\;BSA$ in the prepared conjugates were found to be 9:1 and 5:1, respectively. The titers of the $T_3$ antisera were generally higher (max. $1.5{\times}10^4:1$) than those of $T_3$ (max. $2{\times}10^3:1$), and the average cross reactivity of the $T_3$ antibody with $T_3$ was lower (0.45%) than that of $T_4$ antibody with $T_3(3{\sim}4%)$. The results of the study indicate that the predominant cause of the lower titers and the lower specificity of the $T_4$ antisera comparing with those of $T_3$ is mainly due to the unstability of the $T_4-BSA$ and consequent degradation of the conjugate to $T_3-BSA$ during preparation, purification, and even during immunization. The lower molar ratio of $T_4$ to BSA in the preparation stage is also considered to be a minor factor. By measuring $T_3,\;T_4$ levels in the reference control serum, it has been confirmed that the prepared antisera can sufficiently be utilized, respectively, in the established radioimmunoassay systems.

• 동의생리병리학회지
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• 제17권6호
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• pp.1524-1527
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• 2003

In this study, we demonstrated the antioxidant effect of the Caesalpinia sappan L. extract through the scavenging effect against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and the protective effect on protein damage and PC12 cells against cupric ion/hydrogen peroxide. Its IC/sub 50/ value of the scavenging effect against DPPH radical was 7.7 ㎍. Protection of its extract against oxidative bovine serum albumin (BSA)damage induced by hydrogen peroxide was more effective than that of vitamin C. The protective effect on PC12 cells by hydrogen peroxide was shown to be more potent in is extract than in vitamin C. DNA fragmentation analysis also supports this result.

• BMB Reports
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• 제33권2호
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• pp.133-137
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• 2000

To elucidate the effect of oxygen radicals on the molecular properties of proteins, the secondary and tertiary structure and molecular weight size of BSA and ${\beta}$-lactoglobulin were examined after irradiation of proteins at various doses. Gamma-irradiation of protein solutions caused the disruption of the ordered structure of protein molecules as well as degradation, cross-linking, and aggregation of the polypeptide chains. As a model system, BSA and ${\beta}$-lactoglobulin were used as a typical ${\alpha}$-helical and a ${\beta}$-sheet structure protein, respectively. A circular dichroism study showed that the increase of radiation decreased the ordered structure of proteins with a concurrent increase of aperiodic structure content. Fluorescence spectroscopy indicated that irradiation quenched the emission intensity excited at 280 nm. SDS-PAGE and a gel permeation chromatography study indicated that radiation caused initial fragmentation of proteins resulting in a subsequent aggregation due to cross-linking of protein molecules.

• Macromolecular Research
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• 제17권12호
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• pp.1025-1031
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• 2009

Several kinds of biodegradable hydrogels were prepared via in situ photopolymerization of Pluronic F127/poly($\varepsilon$-caprolactone) macromer and acrylic acid (AA) comonomer in aqueous medium. The swelling kinetics measurements showed that the resultant hydrogels exhibited both thermo- and pH-sensitive behaviors, and that this stimuli-responsiveness underwent a fast reversible process. With increasing pH of the local buffer solutions, the pH sensitivity of the hydrogels was increased, while the temperature sensitivity was decreased. In vitro hydrolytic degradation in the buffer solution (pH 7.4, $37^{\circ}C$), the degradation rate of the hydrogels was greatly improved due to the introduction of the AA comonomer. The in vitro release profiles of bovine serum albumin (BSA) in-situ embedded into the hydrogels were also investigated: the release mechanism of BSA based on the Peppas equation was followed Case II diffusion. Such biodegradable dual-sensitive hydrogel materials may have more advantages as a potentially interesting platform for smart drug delivery carriers and tissue engineering scaffolds.

• 폴리머
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• 제33권1호
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• pp.7-12
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• 2009

PLGA미립구는 주사제형과 같은 형태로 조직공학적 응용에 이용할 수 있다. 본 연구에서는 미립구 제조시 에밀젼 형성조건에 대한 영향과 미립구 표면에 세포를 부착시키는 방법에 대하여 연구하였다. BSA를 함유하는 PLGA미립구는 수중유형(O/W)과 수중유중수형(W/O/W) 용매증발법을 이용하여 제조하였다. 미립구의 초기방출효과제어와 PLGA분해의 과정에서 발생되는 지연시간을 개선시키기 위하며 알긴산나트륨을 수상에 용해시켜 사용하였다. 미립구에 부착된 세포의 형태를 전자주사현미경(SEM)을 이용하여 분석하였고 PLGA미립구에 배양된 인간디스크세포의 증식은 MTT분석을 이용하였으며 이를 통하여 PLGA미립구 표면에 세포가 부착되었음을 확인하였다. 본 연구는 BSA가 함유된 알긴산/PLGA미립구를 이용하여 조직공학적 응용이 가능한 주사제형으로서의 가능성을 제안하였다.

• Journal of Microbiology
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• 제35권2호
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• pp.109-116
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• 1997

An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.

• The Korean Journal of Physiology and Pharmacology
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• 제15권5호
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• pp.299-305
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• 2011

Calcineurin (CaN) is activated in diabetes and plays a role in glomerular hypertrophy and extracellular matrix (ECM) accumulation. Here, kidneys from diabetic model mice were investigated for the expression of the regulator of CaN 1 (RCAN1) isoform 4 (RCAN1.4) which had been shown to be transcriptionally upregulated by CaN activation. We found the increased immunoreactivity for RCAN1 in the glomerular cells of db/db mice and streptozotocin-induced diabetic mice. In concordance, the expression of RCAN1 protein and RCAN1.4 mRNA were elevated in the whole kidney sample from db/db mice. Interleukin-$1{\beta}$ (IL-$1{\beta}$), tumor necrosis factor-${\alpha}$, and glycated albumin (AGE-BSA) were identified as inducers of RCAN1.4 in mesangial cells. Pretreatment of cyclosporine A blocked the increases of RCAN1.4 stimulated by IL-$1{\beta}$ or AGE-BSA, suggesting that activation of CaN is required for the RCAN1.4 induction. Stable transfection of RCAN1.4 in Mes-13 mesangial cells upregulated several factors relevant to ECM production and degradation. These results suggested that RCAN1.4 might act as a link between CaN activation and ECM turnover in diabetic nephropathy.

• Nutritional Sciences
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• 제4권1호
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• pp.26-33
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• 2001

Animal and clinical studies as well as epidemiological data have provided convincing evidence that n-3 polyunsaturated fatty acids can protect against atherosclerosis. However, the effects of the fatty acids on atherogenesis are contradictory. This discrepancy could derive from great susceptibility of the fatty acids to oxidation. We investigated the effect of eicosapentaenoic aced(EPA) on cellular atherogenesis via the scavenger receptor of THP-1 derived macrophages. THP-1 cells were fully differentiated into macrophages by incubating with phorbol 12-myristate 13-acetate for seven days. Atherogenic features of EPA were compared by subsitituting for linoleic acid (LA). Macrophages were also incubated without treatment of the fatty acids as controls. EPA (5-50 nmol/mL) was not cytotoxic and did not measurably induce cellular oxidation compared to bovine serum albumin (BSA) vehicle or identical doses of LA. EPA increased macrophage uptake and degradation of acetylated LDL(AcLDL) up to 14% and 88%, respectively. EPA increased markedly total cellular sterol synthesis and heparin-releasable lipoprotein lipase activity of macrophages, indicating that EPA may enhance accumulation of cellular cholesteryl ester and possibly facilitate formation of foam cells. These results demonstrate that EPA promotes the modified LDL-triggered atherosclerotic process by the modulation of the scavenger receptor and the activation of LPL in macrophages.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권3호
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• pp.179-183
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• 2004

A biocompatible polyelectrolyte complex multilayer (PECML) film consisting of poly-L-lysine (PLL) as a polycation and hyaluronic acid (HA) as a polyanion was developed to test its use for surface modification to prevent cell attachment and protein drug delivery. The formation of PECML through the electrostatic interaction of HA and PLL was confirmed by contact angle measurement, ESCA analysis, and HA content analysis. HA content increased rapidly up to 8 cycles for HA/PLL deposition and then slightly increased with an increasing number of deposition cycle. In vitro release of PLL in the PECML continued up to 4 days and ca. 25% of HA remained on the chitosan-coated cover glass after in vitro release test for 7 days. From the results, PECML of HA and PLL appeared to be stable for about 4 days. The surface modification of the chitosan-coated cover glass with PECML resulted in drastically reduced peripheral blood mononuclear cell (PBMC) attachment. Concerned with its use for protein drug delivery, we confirmed that bovine serum albumin (BSA) as a model protein could be incorporated into the PECML and its release might be triggered by the degradation of HA with hyaluronidase.