• Tuberculosis and Respiratory Diseases
• /
• v.52 no.5
• /
• pp.485-496
• /
• 2002

Background : The NF-${\kappa}B$ transcription factors control various biological processes including the immune response, acute phase reaction and cell cycle regulation. NF-${\kappa}B$ complexes are retained in the cytoplasm in the basal state and various stimuli cause a translocation of the NF-${\kappa}B$ complexes into the nucleus where they bind to the ${\kappa}B$ elements and regulate the transcription of the target genes. Recent reports also suggest that NF-${\kappa}B$ proteins are involved in oncogenesis, tumor growth and metastasis. High expression of NF-${\kappa}B$ expression was reported in many cancer cell lines and tissues. The constitutive activation of NF-${\kappa}B$ was also reported in several cancer cell lines supporting its role in cancer development and survival. The anti-apoptotic action of NF-${\kappa}B$ is important for cancer survival. NF-${\kappa}B$ also controls the expression of several proteins that are important for cellular adhesion (ICAM-1, VCAM-1) suggesting a role in cancer metastasis. In lung cancer, high expression levels of the NF-${\kappa}B$ subunit p50 and c-Rel were reported. In fact, high expression does not mean a high activity, and the activation pattern of NF-${\kappa}B$ in lung cancer has not been reported. Materials and Methods : In this study, the NF-${\kappa}B$ nuclear binding activity in the basal and TNF-${\alpha}$ stimulated states were exmined in various lung cancer cell lines and compared with the normal bronchial epithelial cell line. Twelve lung cancer cell lines including the non-small cell and small cell lung cancer cell lines (A549, NCI-H358, NCI-H441, NCI-H552, NCI-H2009, NCI-H460, NCI-H1229, NCI-H1703, NCI-H157, NCI-H187, NCI-H417, NCI-H526) and BEAS-2B bronchial epithelial cell line were used. To evaluate the NF-${\kappa}B$ expression and DNA binding activity, western blot analysis and an electrophoretic mobility shift assay with the nuclear protein extracts. Results : The basal expressions of the p65 and p50 subunits were observed in the BEAS-2B cell line and all lung cancer cell lines except for NCI-H358 and NCI-H460. The expression levels of p65 and p50 were increased 30 minutes after stimulation with TNF-${\alpha}$ in BEAS-2B and in 10 lung cancer cell lines. In the NCI-H358 and NCI-H460 cell lines, p65 expression was not observed in the basal and stimulated states and the two p50 related protein levels were higher after stimulation with TNF-${\alpha}$ These new proteins were smaller than p50 and are thought to be variants of p50. In the basal state, NF-${\kappa}B$ was nearly activated in the BEAS-2B and all lung cancer cell lines. The DNA binding activity of the NF-${\kappa}B$ complexes was markedly higher after stimulation with TNF-${\alpha}$ In the BEAS-2B and all lung cancer cell line except for NCI-H358 and NCI-H460, the activated NF-${\kappa}B$ complex was a p65/p50 heterodimer. In the NCI-H358 and NCI-H460 lung cancer cell lines, the NF-${\kappa}B$ complex was variant of a p50/p50 homodimer. Conclusion : The NF-${\kappa}B$ activation pattern in the lung cancer cell lines and the normal bronchial epithelial cell lines was similar except for the activation of a variant of the p50/p50 homodimer in some lung cancer cell linse.

• Journal of Life Science
• /
• v.22 no.2
• /
• pp.209-219
• /
• 2012

The properties of lactate dehydrogenase (LDH, EC 1.1.1.27) eye-specific $C_4$ isozyme were studied by polyacrylamide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme kinetics. Furthermore, we proposed the optimal conditions for measuring the activity of LDH eye-specific $C_4$ isozyme. The isozymes were detected in the cytosol of eye tissues from Lepomis macrochirus and Micropterus salmoides and were more similar to the $A_4$ than the $B_4$ isozyme. LDH/CS in the eye tissue of L. macrochirus was increased in September, so the ratio of anaerobic metabolism was high. The electrophoretic patterns of mitochondrial LDH were similar to those of cytosolic LDH in the eye tissues of L. macrochirus and Micropterus salmoides. LDH eye-specific $C_4$ isozyme from eye tissue was purified by preparative native-PAGE. The activities of LDH eye-specific $C_4$ isozymes in L. macrochirus and M. salmoides were reduced at concentrations greater than 0.2 mM and 0.1 mM of pyruvate, respectively. These concentrations remained at 5.2% and 15.8% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The LDH activities of eye tissues were reduced at concentrations greater than 22 mM and 24 mM of lactate, respectively, in L. macrochirus and M. salmoides. The ${K_m}^{PYR}$ of eye-specific $C_4$ was 0.088 mM in L. macrochirus and it was 0.033 mM in M. salmoides. The activities of cytosolic and mitochondrial eye-specific $C_4$ isozymes were high in ${\alpha}$-ketobutyric acid. Furthermore, the activities of eye tissue and eye-specific $C_4$ isozyme had to be measured with 0.5 mM of pyruvate and a buffer solution of pH 7.5. As a conclusion, the eye-specific $C_4$ isozyme in M. salmoides has a high affinity for pyruvate and exhibits maximum activity at a lower concentration of pyruvate and at higher concentration of lactate than that in L. macrochirus. Therefore, it seems that the energy produced by the LDH eye-specific $C_4$ isozyme in M. salmoides was used at the first stage of predatory behavior.

• The Korean Journal of Ecology
• /
• v.16 no.4
• /
• pp.515-526
• /
• 1993

In order to study the ecotypic variation of Rohinia pseudo-acacia L. distributed in southern area of Korean peninsula, 15 local populations(Daejin, Sokcho, Kangneung, Mt. Surak, Hongcheon, Kwangneung, Namhansanseong, Chungju, Yesan, Andong, Jeonju, Dalseong, Changweon, Mokpo and Wando), located from $34^{\circ}18'N\;to\;38^{\circ}36'N$, were selected based on the latitudes and geographical distances. Seeds of these populations were collected and protein contents of seeds and their band patterns were investigated. The seed proteins of all populations were electrophoresed on SDS-polyacrylamide gel. Total number of protein bands were 35, whose molecular weights ranged from 17, 258 daltons to 142, 232 daltons. The number of bands of seed proteins was 23 in Dalseong and Hongcheon and was 32 in Daejin and Sokcho, showing an increasing tendency in the number of bands as the latitude goes high. The local populations were classified into 3 local types based on protein analysis: the middle north east coastal type(Daejin, Sokcho. Kangneung), the central type (Mt. Surak, Hongcheon, Kwangneung, Namhansanseong, Chungju) and the southern type(Yesan, Andong, Jeonju, Dalseong, Changweon, Mokpo, Wando). According to the results of cluster analysis by UPGMA based on the similarity index(c0efficient of Jaccard) of the patterns, 3 local types were subdivided further into 6 types: the middle north east coastal type(Sokcho, Kangneung), the north central type I (Mt. Surak, Hongcheon), the north central type II (Narnhansanseong, Chungju, Daejin), the north central type III (Kwangneung), the south central type (Yesan, Dalseong, Jeonju) and the southern type(Andong, Changweon, Mokpo, Dalseong, Wando). The No. 12 band of the separated seed proteins showed the highest colored density in the preparations from all the populations. The No. 11~13 and No. 23~28 bands also showed high densities. As a whole, southern type populations (Changweon, Mokpo, Wando) showed high protein contents and high colored density. Total protein contents of the seeds in each population were variable from 9. 68mg / g (Mt. Surak) to 17.30mg/g (Jeonju), showing an increasing trends toward low latitudes.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.12 no.1
• /
• pp.46-50
• /
• 1983

In order to find out the possibility of utilizing red pepper seed as food resources of fats and proteins, a series of studies were conducted. The red pepper seed contained 27.6% of crude fat and 22.2% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.4% neutral lipid, where as compound lipid were 4.6%. Among the neutral lipid separated by thin layer chromatography, triglyceride was 85.6%, sterol ester 4.9%, free fatty acids 3.4%, diglyceride 2.5%, sterol 2.2% and monoglyceride 1.1%, respectively. The predominant fatty acids of red pepper seed oil were linoleic acid (57.1-75.4%), palmitic acid (13.9-21.3%) and oleic acid (8.0-15.1%), especially glycolipid contained 1.7% of linolenic acid and small amount of myristic acid and arachidic acid. The salt soluble protein of red pepper seed was highly dispersible in 0.02M sodium phosphate buffer containing 1.0M $MgSO_4$, and the extractability of seed protein was about 25.0%. Glutamic acid and arginine were major amino acids of red pepper seed protein. The electrophoretic analysis showed 6 bands in seed protein, and the collection rate of the main protein fraction purified by sephadex G-100 and G-200 was about 62.2%. Glutamic acid (19.9%) was major amino acid of the main protein, followed by glycine and alanine. The molecular weight of the main protein was estimated to be 93,000.

• Korean Journal of Food Science and Technology
• /
• v.14 no.3
• /
• pp.250-256
• /
• 1982

A series of studies were conducted to find out the possibility of utilizing grape seed as resources of food fats and proteins, and the results of the studies are as follows: The grape seed contained 25.1%, of crude fat and 12.0% of crude protein. The lipid, fractions obtained by silicic acid column chromatography were mainly composed of about 95.5% neutral lipid, whereas compound lipid was only 4.5% level. Among the neutral lipid by thin layer chromatography, triglyceride was 91.89%, sterol ester, sterol, diglyceride and free fatty acid were 3.24%, 2.87%, 1.20% and 0.80%, respectively The predominant fatty acids of total and neutral lipids were linoleic acid $(69.72{\sim}71.72%)$ and oleic acid $18.09{\sim}19.46%)$, but those of glycolipid and phospolipid were linoleic acid $(31.49{\sim}38.18%)$, oleic acid $(20.20{\sim}35.27%)$ and palmitic acid $(26.80{\sim}39.98%)$. The major fatty acids of triglyceride separated from neutral lipid were oleic acid (43.08%), linoleic acid (38.42%) and palmitic acid (11.60%). The salt soluble protein of grape seed was highly dispersible in 0.02M sodium phosphate buffer containing about 1.0M $MgSO_4$, and the extractability of seed protein was 31%. Glutamic acid was the major amino acid in salt soluble protein, followed by arginine and aspartic acid. The electrophoretic analysis showed 3 bands in grape seed protein, and the collection rate of the main protein fraction purified by Sephadex G-100 and G-200 was 82%. Glutamic acid, aspartic acid and arginine were the major amino acids of the main grape seed protein. The molecular weight for the main protein of the grape seed was estimated to be 81,000.

• Journal of Dairy Science and Biotechnology
• /
• v.27 no.2
• /
• pp.7-16
• /
• 2009

Appenzeller cheese samples were prepared by addition of 0.5, 1.0, and 2.0% green tea (Camellia sinensis, CS) powder and control cheese. We examined various quality characteristics of the novel cheese, such as viable-cell counts, pH, water-soluble nitrogen (WSN), non-casein nitrogen (NCN), non-protein nitrogen (NPN), and catechin level during maturation for 16 weeks at $14^{\circ}C$. To develop a Korean natural cheese containing green tea powder, we also analyzed the changes in the polyacrylamide gel electrophoresis pattern, chemical composition, and sensory qualities. The viable cell counts of the samples were not significantly different. Until the $3^{rd}$ week, the pH of the CS cheese decreased with an increase in the maturation time. However, the pH gradually increased by the $12^{th}$ week, while WSN, NCN, NPN also increased. The WSN, NCN, NPN, and catechin values for the CS cheese samples were significantly higher than the values for the control cheese. The polyacrylamide gel electrophoretic pattern of caseins for the CS cheese indicated that this cheese degraded more rapidly than the control cheese did. In the sensory evaluation, cheese with 1.0% CS powder showed the highest scores in taste and appearance and good scores in flavor and texture. These results indicate that 1.0% CS is the optimal value for addition to cheese, and cheese containing 1.0% CS shows good physiological properties and reasonably high overall sensory acceptability.

• Korean Journal of Environmental Agriculture
• /
• v.25 no.1
• /
• pp.7-13
• /
• 2006

Plastichouse cultivation for crops and vegetables in the winter has been widely popularized in Korea. In the vinylhouse Ultraviolet B penetration is lower than in the field, and so some problems, as plant overgrowth and outbreak of disease, occurred frequently. The effect of artificial supplement ultraviolet B $(UV-B:280{\sim}320nm)$ radiation on the physiological responses and yield of perilla (perilla frutescens) was investigated UV-B ray was radiated on perilla with the 10th leaf stage at the distance of 90, 120 and 150 cm from the plant canopy for 30 days after planting in the vinylhouse. The production of fresh perilla leaves was high in the order of plastic house, ambient+50% of supplemental UV-B, ambient ambient+100% of supplemental UV-B. Enhanced UV-B radiation affected the intensity of thirty-three proteins in 2-dimensional electrophoretic analysis of proteins and ten proteins out of them seemed to be responsive to UV-B : a protein was, ATP synthase CF1 alpha chain, down regulated and nine proteins (Chlorophyll a/b bindng protein type I, Chlorophyll a/b binding protein type II precursor, Photosystem I P700 chlorophyll a apoprotein A2, DNA recombination and repair protein recF, Galactinol synthase, S-adenosyl-L-methionine, Heat shock protein 21, Calcium-dependent protein kinase(CDPK)-like, Catalase) were up-regulated.

• The Korean Journal of Pharmacology
• /
• v.30 no.1
• /
• pp.125-136
• /
• 1994

To characterize the SR Ca-release channel protein complex of crustacean, $^{45}Ca-release,\;[^3H]ryanodine$ binding, and immunoblot studies were carried out in the crayfish and/or lobster skeletal sarcoplasmic reticulum. Bmax and affinity of crayfish SR to ryanodine were lower than those of lobster SR. AMP (5mM) increased $[^3H]ryanodine$ binding significantly in both vesicles (P<0.05). $Mg^{2+}$(5mM) or tetracaine(1mM) inhibited $[^3H]ryanodine$ binding significantly in both vesicles (P<0.001), but ruthenium red $(10\;{\mu}M)$ inhibited it moderately. In SDS polyacrylamide gel electrophoretic analysis of crayfish SR vesicles, there was a high molecular weight band that showed similar mobility with Ca-release channel protein of lobster skeletal SR, but more rapid mobility (HMWBr) than that of rabbit skeletal SR (HMWBS). Immunoblot analysis showed that polyclonal Ab to lobster skeletal SR Ca-release channel protein was react with HMWBr of crayfish skeletal SR, but not with that of HMWBs of rabbit skeletal SR. ^{45}Ca-release from crayfish skeletal SR vesicles was increased by the increase of extravesicular calcium from $1{\mu}M$ to 1mM. This Ca-release phenomenon was similar, but more sensitive in the low concentration of $Ca^{2+}$, compared to that from lobster SR vesicles. AMP (5mM) or caffeine (10mM) did not affect to $^{45}Ca-release.\;^{45}Ca-release$ was inhibited slightly ($3{\sim}8%\;by\;Mg^{2+}$) (5mM) or tetracaine (1mM), and moderately (23%) by high concentration of ruthenium red $(300\;{\mu}M)$. From the above results, it is suggested that SR Ca-release channel protein of crustacean has different properties from that of the rabbit, and similar properties between crayfish and lobster in functional and immunological aspects, but Ca-release via crayfish channel may be more sensitive to calcium.

• The Korean Journal of Pharmacology
• /
• v.29 no.2
• /
• pp.275-282
• /
• 1993

Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.

• Korean Journal of Agricultural Science
• /
• v.5 no.2
• /
• pp.56-67
• /
• 1978

This study was conducted to characterize comparatively the accumulative patterns of protein and oil, temporal changes in electrophoretic components of proteins during seed development and maturation for the soybean varieties with high, medium and low protein contents. 1. The dry matter of the developing seed increases slowly for the first 22 days after flowering, followed by rapid linear increase for 20 to 30 days and further slow increase for 5 to 15 days attaining its maximum. During the period 12 to 27 days after flowering the protein content of seed increases rapidly while oil content increases rapidly. Following this period of rapid changes, there was period of slow increase until 40 to 47 days after flowering and no seizable further change in the content of both protein and oil. 2. The high protein variety, Saikai # 20, was characterized by shorter period and lower rate of decrease in protein content during the early period, followed by longer period and higher rate of increase in protein content, with earlier stop of oil accumlation during the seed development. 3. The low protein and high oil variety, Shelby, was characterized by longer period of decrease in protein content and shorter period of increase in protein content in contrast to the longer period of slow oil increase during seed development. 4. The temporal pattern of protein component accumulation during seed development was distinctly different among varieties differing in protein content. The time of distinct appearance of all the protein components identifiable in the matured seeds was in accordance with the end of d crease in the protein content of seed. A component having Rm of 0.03 which was absent in the matured seeds was identifiable during the first 17 days after flowering. 5. The high protein variety, Saikai # 20, had much higher compositioral ratio of the component a from the early days of seed development and it continued to increase until 47 days after flowering, while the increase in the composition of the component a stopped as early as 27 days after flowering in the other lower protein varieties. 6. The composition of the component b increased during the period from 17 to 42 days after flowering in all the varieties tested, but the rate of increase during the period was lowest in the high protein variety, Saikai # 20.