• Food Science and Preservation
• /
• v.19 no.2
• /
• pp.216-222
• /
• 2012

The optimum annealing conditions of corn starch slurry were studied for RS4 type resistant starch production by phosphorylated cross-linking. When a corn starch slurry was cross-linked by using phosphate salts (STMP/STPP mixture) in the presence of 0.9%, 1.2% and 1.5% NaOH/st.ds, a high concentration of NaOH resulted in a rapid increase of the RS contents at the early reaction stage. However, similar RS contents were obtained after 12 h of cross-linking regardless of NaOH concentrations. The annealing treatment was conducted under various conditions such as pH between 2-10, temperature $40-60^{\circ}C$, time 0-14 h followed by phosphorylated cross-linking. The lower slurry pH was for the annealing treatment, the higher RS contents were obtained after cross-linking. When the slurry annealed for various period of time and temperature, a maximal amount of RS was formed after 2 h of annealing at $50^{\circ}C$ of annealing temperature of the starch slurry (pH 2.0). Therefore, an optimal annealing conditions at pH 2.0 and $50^{\circ}C$ for 2 h were proposed under the cross-linking conditions of sodium sulfate 10%/st.ds, NaOH 1.2%/st.ds and 12 h of the reaction time. The RS contents were linearly increased with the increase of phosphate salt addition. The RS4 prepared under the optimal conditions contained RS 72.3% and its phosphorus content was 0.36%/st.ds, which was below the limit (0.4%/st.ds) of modified starch by Korea Food Additives Code.

• Toxicological Research
• /
• v.8 no.2
• /
• pp.343-357
• /
• 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.