• Toxicological Research
• /
• v.4 no.1
• /
• pp.37-45
• /
• 1988

Primary cultures of adult rat hepatocytes were used to study in vitro cytotoxic effects of T-2 toxin on liver cells. When T-2 toxin was added to the culture, a significant depression of the hormonal induction of ${\alpha}$-aminoisobutyric acid (AIB) uptake and tyrosine aminotransferase (TAT) activity was observed. However, T-2 toxin did not affect the uptake of ouabain into hepatocytes. Protein synthesis was inhibited by T-2 toxin, but RNA synthesis was not severely affected. The inhibitory effects of T-2 toxin on protein synthesis was diminished rapidly with culture time and the hepatocytes culture maintained control level of protein synthesis within 24 hrs.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.9
• /
• pp.2607-2612
• /
• 2010

Human tissue-type plasminogen activator (tPA) is a valuable thrombolytic agent used to successfully treat acute myocardial infarction, thromboembolic stroke, peripheral arterial occlusion, and venous thromboembolism. Recombinant tPA is accumulated as an inactive form in inclusion bodies of E. coli and is refolded in vitro, which is accompanied by extensive aggregation. In the present study, a tPA protease domain was expressed in an active soluble form in the cytosol of E. coli Rosetta-gami cells, which allowed disulfide bond formation and supplied the tRNA molecules required for six rarely used codons in E. coli. This strategy increased the amount of soluble protease domain protein and avoided the cumbersome refolding process. The purified protease domain not only degraded tPA substrate peptides but also formed a covalently bound complex with plasminogen activator inhibitor-1, as does full-length tPA. Soluble expression and purification of tPA domains may aid in functional analyses of this multi-domain protein, which has been implicated in many physiological and pathological processes.

• The Korean Journal of Zoology
• /
• v.26 no.3
• /
• pp.181-192
• /
• 1983

Cytosol protein patterns of two strains of A. proteus, tD and xD strain, were compared by two dimensional gel electrophoresis. Among the 200 major polypeptides that could be stained by silver stain method, tD strain contained a cell specific protein whose molecular weight was 45,000 dalton, pI 5.9. On the other hand, the cytosol and the symbiotic vesicles of xD strain contained a symbiosis specific protein (M.W. 29,000; pI 5.5). The fate of the symbiosis specific protein depended on the presence of symbiotic bacteria in the experiment of high temperature effect and of experimental infection. The significance of these results is discussed in relation to their function in organismic association on the basis of the previous findings.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.8
• /
• pp.1062-1068
• /
• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

• Nutrition Research and Practice
• /
• v.9 no.4
• /
• pp.343-349
• /
• 2015

BACKGROUND/OBJECTIVES: Fermentation of dietary fiber results in production of various short chain fatty acids in the colon. In particular, butyrate is reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene expression or the number of goblet cells. The objective of this study was to investigate whether butyrate modulates mucin secretion in LS174T human colorectal cells, thereby influencing the adhesion of probiotics such as Lactobacillus and Bifidobacterium strains and subsequently inhibiting pathogenic bacteria such as E. coli. In addition, possible signaling pathways involved in mucin gene regulation induced by butyrate treatment were also investigated. MATERIALS/METHODS: Mucin protein content assay and periodic acid-Schiff (PAS) staining were performed in LS174T cells treated with butyrate at various concentrations. Effects of butyrate on the ability of probiotics to adhere to LS174T cells and their competition with E. coli strains were examined. Real time polymerase chain reaction for mucin gene expression and Taqman array 96-well fast plate-based pathway analysis were performed on butyrate-treated LS174T cells. RESULTS: Treatment with butyrate resulted in a dose-dependent increase in mucin protein contents in LS174T cells with peak effects at 6 or 9 mM, which was further confirmed by PAS staining. Increase in mucin protein contents resulted in elevated adherence of probiotics, which subsequently reduced the adherent ability of E. coli. Treatment with butyrate also increased transcriptional levels of MUC3, MUC4, and MUC12, which was accompanied by higher gene expressions of signaling kinases and transcription factors involved in mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: Based on our results, butyrate is an effective regulator of modulation of mucin protein production at the transcriptional and translational levels, resulting in changes in the adherence of gut microflora. Butyrate potentially stimulates the MAPK signaling pathway in intestinal cells, which is positively correlated with gut defense.

• Food Science of Animal Resources
• /
• v.34 no.2
• /
• pp.192-199
• /
• 2014

The effects of adding mechanically deboned chicken (MDC) hydrolysates on the quality characteristics of imitation crab stick (ICS) during storage were investigated. ICS was prepared from Alaska Pollack, chicken breast surimi, and protein hydrolysates enzymatically extracted from MDC. ICS samples were divided into 4 groups: without protein hydrolysate (control), added with 0.5% protein hydrolysate (T1), added with 1.0% protein hydrolysate (T2), and added with 1.5% protein hydrolysate (T3). Results showed that crude protein content did not differ significantly among the ICS samples (p>0.05). ICS sample added with MDC hydrolysates had higher crude fat and ash content but lower moisture content than the control (p<0.05). Lightness was significantly lower in T2 and T3 than in the other groups at 0 and 4 wk of storage. Also, whiteness decreased in the groups contained MDC hydrolysates. Breaking force and jelly strength were higher in samples containing MDC hydrolysates compared to control samples (p<0.05). Additionally, saturated fatty acid contents were lower in the groups containing MDC hydrolysates than in control sample groups (p<0.05). Polyunsaturated fatty acid (PUFA) and essential fatty acids (EFA) were significantly higher in T2 and T3 than the control samples. In particular, all samples containing MDC hydrolysates had reduced thiobarbituric acid-reactive substances (TBARS) values at 4 wk. Free radical scavenging activity also was increased with addition of MDC hydrolysates.

• Molecules and Cells
• /
• v.24 no.3
• /
• pp.437-440
• /
• 2007

$OGL-20P^T$-358 is a novel 66 amino acid residue protein from the hyperthermophile Thermococcus thioreducens sp. nov., strain $OGL-20P^T$, which was collected from the wall of the hydrothermal black smoker in the Rainbow Vent along the mid-Atlantic ridge. This protein, which has no detectable sequence homology with proteins or domains of known function, has a calculated pI of 4.76 and a molecular mass of 8.2 kDa. We report here the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of $OGL-20P^T$-358. Assignments are 97.5% (316/324) complete. Chemical shift index was used to determine the secondary structure of the protein, which appears to consist of primarily ${\alpha}$-helical regions. This work is the foundation for future studies to determine the three-dimensional solution structure of the protein.

• Transactions of the Society of Information Storage Systems
• /
• v.9 no.1
• /
• pp.28-31
• /
• 2013

We proposed a method to fabricate label-free protein sensor with sub-wavelength nanograting structures to be used for diagnosing acute myocardial infarction. A nickel stamp for the injection molding of nanograting integrated protein sensor was fabricated by electroforming process with high fidelity. By using metallic stamp, we replicated label-free protein sensor via injection molding, which is an outstanding method for low-cost and mass production of polymer products. Finally, we performed a feasibility test, examining cardiac troponin T (cTnT) and anti-cTnT interactions. From the results, we demonstrated that the fabricated protein sensor can provide information for the early and accurate detection of cardiac diseases such as acute myocardial infarction.

• Clinical and Experimental Pediatrics
• /
• v.45 no.11
• /
• pp.1368-1372
• /
• 2002

Purpose : Group A streptococci have a cell wall which consists of M protein and T protein. T protein is known to be helpful in the understanding of the epidemiology of group A streptococci. To study the epidemiologic characteristics, we serotyped T protein of group A streptococci obtained from patients admitted to hospitals, or who visited OPD in five districts of Seoul the during last three years. Methods : Group A streptococci were obtained in five districts in north, northeast, central, northwest and south Seoul from 1998 through 2000. All isolated group A streptococci were serotyped with T protein antisera(Institute of Sera and Vaccine, Prague, Czech Republic). Results : In 1998, analysis of obtained total number of 92 strains revealed that T12, T4, and NT acounted for 72.2%. Among seven cases of scarlet fever, T12 was isolated in four cases and T4 was found in three cases. Two cases of tonsilar abscess produced T8 and NT. One case of cervical lymphadenitis showed T12. In 1999, 41 cases were studied showing that T12, T4, and T1 contributed 68%. Among five cases of scarlet fever, T12 and T4 make up three case. There were two cases of pneumonia(T4 and T1) and one case of cervical lymphadenitis(T8/25). In 2000, the study was performed in four districts except the central area. Among 83 isolates, T12, T4 and T1 accounted for 63.9%. There were three cases of scarlet fever(T12, T4, T5), one case of tonsillar abscess(T12), one case of pneumonia(NT) and one case of sepsis(T1). Conclusion : Serological analysis of T protein of group A streptococci shows no endemic specificity. The yearly pattern reveals that T12 had been decreasing but T1 had shown the opposite trend.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.8
• /
• pp.1186-1194
• /
• 2019

Objective: The influence of broiler carcass scalding conditions on chicken breast meat quality parameters was investigated. Methods: Two hundred and seventy Cobb broiler chickens from 42 to 48 days old were slaughtered according to the standard industry practice and scalded in five temperature/time combinations-$T_1$, $54^{\circ}C/210s$; $T_2$, $55^{\circ}C/180s$; $T_3$, $56^{\circ}C/150s$; $T_4$, $57^{\circ}C/120s$; $T_5$, $58^{\circ}C/90s$. Results: Scalding temperature increase resulted in higher values of external and ventral lightness and in protein functionality reduction-determined by emulsification capacity and protein denaturation-in chicken breast fillets 24 h post-mortem. Protein secondary structures had conformational changes, with a decrease of the ${\alpha}$-helix and an increase of the ${\beta}$-sheet and ${\beta}$-turn proportions, mainly in $T_1$ and $T_5$ samples, determined by Fourier-transform infrared spectroscopy in an attenuated reflectance mode analysis. The chemical composition, pH, water holding capacity and Warner-Bratzler shear force did not differ among the treatments. In the fatty acid profile, the 18:1n-9 was lower in $T_5$, which suggested that the high scalding-temperature could have caused the lipid oxidation. The values of the polyunsaturated fatty acids (PUFA), such as 22:2, 20:4n-6, and 22:6n-3, were highest in the $T_5$, thus being related to the phospholipid cellular membrane collapse in this experimental condition and subsequent release of these PUFA. Conclusion: Intermediate scalding-parameters avoided the negative changes in the chicken meat quality.