• Mass Spectrometry Letters
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• v.1 no.1
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• pp.25-28
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• 2010

The high-throughput identification and accurate quantification of proteins are essential strategies for exploring cellular functions and processes in quantitative proteomics. Stable isotope tagging is a key technique in quantitative proteomic research, accompanied by automated tandem mass spectrometry. For the differential proteome analysis of mouse neuronal cell lines, we used a multiplexed isobaric tagging method, in which a four-plex set of amine-reactive isobaric tags are available for peptide derivatization. Using the four-plex set of isobaric tag for relative and absolute quantitation (iTRAQ) reagents, we analyzed the differential proteome in several stroke time pathways (0, 4, and 8 h) after the mouse neuronal cells have been stressed using a glutamate oxidant. In order to obtain a list of the differentially expressed proteins, we selected those proteins which had apparently changed significantly during the stress test. With 95% of the peptides showing only a small variation in quantity before and after the test, we obtained a list of eight up-regulated and four down-regulated proteins for the stroke time pathways. To validate the iTRAQ approach, we studied the use of oxidant stresses for mouse neuronal cell samples that have shown differential proteome in several stroke time pathways (0, 4, and 8 h). Results suggest that histone H1 might be the key protein in the oxidative injury caused by glutamate-induced cytotoxicity in HT22 cells.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.4
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• pp.243-251
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• 2006

Dideoxypetrosynol A, a polyacetylene from the marine sponge Petrosia sp., is known to exhibit significant selective cytotoxic activity against a small panel of human tumor cell lines, however, the mechanisms of which are poorly understood. In the present study, it was investigated the further possible mechanisms by which dideoxytetrosynol A exerts its anti-proliferative action in cultured human leukemia cell line U937. We observed that the proliferation-inhibitory effect of dideoxypetrosynol A was due to the induction of G1 arrest of the cell cycle and apoptosis, which effects were associated with up-regulation of cyclin D1 and down-regulation of cyclin E without any change in cyclin-dependent-kinases (Cdks) expression. Dideoxypetrosynol A markedly induced the levels of Cdk inhibitor p16/INK4a expression. Furthermore, down-regulation of phosphorylation of retinoblastoma protein (pRB) by this compound was associated with enhanced binding of pRB and the transcription factor E2F-1. The increase in apoptosis was associated with a dose-dependent up-regulation in pro-apoptotic Bax expression and activation of caspase-3 and caspase-9. Dideoxytetrosynol A decreased the levels of cyclooxygenase (COX)-2 mRNA and protein expression without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin E2 (PGE2) synthesis. Furthermore, dideoxytetrosynol A treatment markedly inhibited the activity of telomerase, and the expression of human telomerase reverse transcriptase (hTERT), a main determinant of the telomerase enzymatic activity, was progressively down-regulated by dideoxytetrosynol A treatment in a dose-dependent fashion. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of dideoxytetrosynol A.

• Journal of the Korean Dietetic Association
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• v.8 no.3
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• pp.301-310
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• 2002

In order to investigate the use and recognition of nutrition labelings of processed foods which adolescents frequently purchase, 277(boys 125, girls 152) middle school students and their parents were conveniently selected from the west Chonan City, and were surveyed by self-recording questionnaire on June, 2000. Nutrition labels were indicated only in 14.9% of 222 processed snack foods. At purchasing foods the students considered taste most importantly and in turn price, quantity, and nutrition. Of the subjects 79.4% of students and 89.2% of parents had experiences of reading nutrition labels, and the reasons were mainly to just know contained nutrients(76.8%, 72.1%, respectively) and for their health care(35.9%, 47.8%, respectively). The main reason not to confirm nutrition labels was that they were not interested in nutrition labels. But too small size and rudeness or hardness of understanding was also pointed out as a reason of not reading nutrition labels. The students ranked mineral, protein, and carbohydrate as the most important three nutrients at labelings, while the parents ranked protein, mineral, and calorie. Cholesterol, dietary fibers, and sodium were shown the lowest rank in both students and parents. Significant nutrients for specific processed foods did not well recognised in the students. Conclusively, nutrition labelings should be applied to all processed foods which students frequently buy, particularly in clear and large size. Also it is necessary to prepare education programs for both students and parents about accurate recognition of nutrition knowledge about processed foods and how to use nutrition labels.

• Animal cells and systems
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• v.10 no.4
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• pp.227-231
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• 2006

Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

• Korean Journal of Veterinary Research
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• v.49 no.1
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• pp.1-8
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• 2009

The ubiquitin-proteasome mediated protein degradation pathway plays an important role in regulating both cell proliferation and cell death. Proteasome inhibitors are well known to induce apoptosis in various human cancer cell lines. We investigated the effect of combined treatment with proteasome inhibitor and TRAIL, and a possible mechanism of the enhancing apoptosis by the both treatment, on TRAIL-resistant non-small cell lung cancer. A549 cells were exposed to the N-Acetyl-Leu-Leu-Norleu-al (ALLN) as a proteasome inhibitor and then treated with recombinant TRAIL protein. In A549 cells under proteasome inhibition conditions by pretreatment with ALLN, TRAIL treatment significantly decreased cell viability compared to that ALLN and TRAIL alone treatment. Also, the both treatment induced cell damage through DNA fragmentation and p53 expression. In addition, the combined treatment of both markedly increased caspase-8 activation, especially the exposure for 2 h, and Bax expression and induced the dissipation of mitochondrial transmembrane potential in A549 cells. Taken together, these findings showed that proteasome inhibition by ALLN enhanced TRAIL-induced apoptosis via DNA degradation by activated P53 and mitochondrial transmembrane potential loss by caspase-8 activation and bax expression. Therefore, our results suggest that proteasome inhibitor may be used a very effectively chemotherapeutic agent for the tumor treatment, especially TRAIL-resistant tumor cell.

• Food Science of Animal Resources
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• v.34 no.3
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• pp.297-306
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• 2014

Basic information regarding the yield and nutritional composition of edible pork by-products, namely heart, liver, lung, stomach, spleen, uterus, pancreas, and small and large intestines, was studied. Our results revealed that the yields varied widely among the pork by-products examined; in particular, liver had the highest yield (1.35%); whereas, spleen had the lowest yield (0.16%). The approximate composition range (minimum to maximum) of these by-products was found to be: moisture 71.59-82.48%; fat 0.28-19.54%; ash 0.155-1.34%, and protein 8.45-22.05%. The highest protein, vitamin A, B2, B6, and total essential amino acid (EAA) contents were found in liver. Large intestine had the highest fat content and lowest EAA content. Heart had the highest vitamin B1 content, whereas pancreas had the highest niacin and vitamin B3 contents. The concentrations of Fe and Zn were highest in liver and pancreas. Total saturated fatty acids (SFA) levels and polyunsaturated fatty acids (PUFA) levels between the by-products ranged from 43.15-50.48%, and 14.92-30.16%, respectively. Furthermore, with the exception of large intestine, all the by-products showed favorable PUFA/SFA ratios. The study indicated that almost all of the pork by-products examined were good sources of important nutrients, and that these data will be of great importance in the promotion of the consumption of edible pork by-products, as well as their utilization in meat processing.

• Archives of Pharmacal Research
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• v.19 no.5
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• pp.359-367
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• 1996

The pharmacokinetics of LB20304 was investigated following intravenous (IV) and oral administration to rats and dogs. Additionally, in vitro metabolism and serum protein binding studies were also conducted. The total body clearance, apparent volume of distribution, terminal half-life, and extent of bioavailability were 21.8 ml/min/kg, 2265 ml/kg, 93.6 min, and 30.8% for rats; and 7.95 ml/min/kg, 4144 ml/kg, 363 min, and 81.1% for dogs, respectively. LB20304 was stable in the liver microsome containing NADPH generating system and its serum protein binding was 58.5-65.8% for rats, 19.1-29.6% for dogs, and 56.9-59.6% for humans. Its tissue concentration levels in lever, stomach, small intestine, and kidney were 9.5 to 26.1 times greater than plasma level, but the concentration in testis was quite low and that in brain was negligible in rats. The 48 hr urinary recovery of the dose was 44% for IV dosing and 14% for oral dosing, shereas the 48 hr biliary recovery of the dose was 6.4% for IV dosing and 4.5% for oral dosing in rats. In summary, the pharmacokinetic properties of LB20304 were characterized by its good oral absorption, long plasma half-life, and good tissue distribution.

• Korean Journal of Community Nutrition
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• v.7 no.2
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• pp.277-285
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• 2002

The purpose of this study was to develop a semi-quantitative food frequency questionnaire (SQ-FFQ) for subjects aged 50yr and over and to evaluate the validity of this SQ-FFQ. Dietary intake was assessed using SQ-FFQ that included 98 commonly consumed flood items selected from the results of the Korean Health and Nutritional Survey, 1998. Subjects (n = 2,660) aged 50yr and over were recruited from 7 metropolitan cities and 8 small cities. Each subject was interviewed using this SQ-FFQ developed in our laboratory and 24hr-recall method. Excluding incomplete data, Data from 1,149 subjects were used in this validity study. The nutrient intakes assessed by this SQ-FFQ were validated by comparing with the results from 1 day 24-hour recalls. Pearson's correlation coefficients between two methods were 0.71, 0.64, 0.53, and 0.43 for energy, carbohydrate, protein, and fat, respectively for all subjects. Spearman's correlation coefficients were higher than those of Pearson's correlation coefficients. Kappa values for energy, carbohydrate, protein, and fat were 0.79, 0.72, 0.70, and 0.64, respectively. The percentage for misclassification of the lowest quartile into the highest quartile or vice versa was 1.25-1.39% for all nutrients. Therefore, this SQ-FFQ seems to be useful in assessing the nutritional status of the middle-aged and elderly subjects in Korea.

• BMB Reports
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• v.29 no.6
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• pp.549-554
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• 1996

Phosphoinositide turnover in response to platelet-derived growth factor, epidermal growth factor, and bradykinin was evaluated in NIH 3T3 cells. Platelet-derived growth factor and bradykinin induced a significant increase in incorporation of $^{32}P$ into phosphatidylinositol (PI), phosphatidylinositol 4-monophosphate (PIP), and phosphatidylinositol 4.5-bisphosphate ($PIP_2$) in serum-starved NIH 3T3 cells. However, epidermal growth factor increased incorporation of $^{32}P$ into these phosphoinositides by only a small amount. Stimulation with platelet-derived growth factor, not bradykinin, caused a rapid elevation of PI and PIP kinase activities that were maximally activated within 10 min. The maximal levels of their elevation in cells with plateletderived growth factor stimulation were 3.2-fold for PI kinase, and 2.1-fold for PIP kinase. Short term pretreatment of NIH 3T3 cells with phorbol 12-myristate 13-acetate, activator of protein kinase C. caused an approximately 60% decrease in platelet-derived growth factor-induced PI kinase activities, indicating the feedback regulation of phosphoinositide turnover by protein kinase C. These results suggest that although the enhancement of phosphoinositide turnover is a rapidly occurring response in platelet-derived growth factor- or bradykinin-stimulated NIH 3T3 cells, phosphoinositide kinases may be associated with initial signal transduction pathway relevant to platelet-derived growth factor but not to bradykinin.

• YAKHAK HOEJI
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• v.57 no.4
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• pp.250-257
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• 2013

Biosimilars are important drugs in medicine and contain many glycosylated proteins. Thorough analysis of the glycosylated protein is a prerequisite for evaluation of biosimilar glycan drugs. A method to assess the diversity of N-glycosylation sites and N-glycans from biosimilar glycan drugs has been developed using two separate methods, LC-MS/MS and MALDI-TOF MS, respectively. Development of sensitive, accurate, and efficient methods for evaluation of glycoproteins is still needed. In this study, analysis of both N-glycosylation sites and N-glycans of glycoprotein was performed using the same LC-MS/MS with two different nano-LC columns, nano-C18 and nano-porous graphitized carbon (nano-PGC) columns. N-glycosylated proteins, including RNAse B (one N-glycosylation site), Fetuin (three sites), and ${\alpha}$-1 acid glycoprotein (four sites), were used, and small amounts of each protein were used for identification of N-glycosylation sites. In addition, high mannose N-glycans (one type of typical glycan structure), Mannose 5 and 9, eluted from RNAse B, were successfully identified using nano-PGC-LC MS/MS analysis, and the abundance of each glycan from the glycoprotein was calculated. This study demonstrated an accurate and efficient method for determination of N-glycosylation sites and N-glycans of glycoproteins based on high sensitive LC-MS/MS using two different nano-columns; this method could be applied for evaluation of the quality of various biosimilar drugs containing N-glycosylation groups.