• 한국식품저장유통학회지
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• 제19권2호
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• pp.216-222
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• 2012

옥수수 전분의 인산화 가교반응에 의한 RS4 제조 시 최적 annealing 조건을 검토하였다. 전분유에 NaOH를 0.9%, 1.2%, 1.5%/st.ds의 농도가 되게 첨가하고 복합인산염(STMP/STPP 혼합)을 이용하여 인산화 가교반응 할 경우 NaOH 농도가 높을수록 반응 초기에 RS 함량이 빠르게 증가되었으나 NaOH 농도에 상관없이 반응 12시간 후에는 비슷한 RS 함량을 나타내었다. 전분유의 Annealing 처리는 인산화 가교반응 전에 실시하였다. 전분유를 pH 2-10, 온도 $40-60^{\circ}C$, 0-14시간으로 처리한 후 인산화 가교반응을 하였다. Annealing 처리시 pH가 낮을수록 RS 함량이 높아지는 결과를 보였으며 pH 2에서 annealing 온도 $50^{\circ}C$, 2시간 annealing에서 최고의 RS 함량을 나타내었다. 따라서 최적 인산화 가교반응 조건은 먼저 pH 2.0, $50^{\circ}C$에서 2시간 annealing 처리를 한 후 황산나트륨 농도 10%/st.ds, NaOH 농도 1.2%/st.ds 조건에서 12시간 인산화 가교반응을 행하는 것으로 생각된다. RS 함량은 인산염 투입량이 증가함에 따라 직선적으로 증가하였다. 본 연구결과에 의한 최적조건을 활용하여 복합인산염 10%/st.ds의 농도로 RS4를 제조한 결과 RS 함량은 72.3% 였으며 인의 함량은 0.36%/st.ds로 한국 식품첨가물 공전에 적합하여 식품용 소재로 활용이 가능할 것으로 사료된다.

• Toxicological Research
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• 제8권2호
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.