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.
Changes of protein and amino acids composition in improvement-Meju inoculated with Aspergillus oryzae were emamined at various time intervals over 6-day test period. To investigate those changes systematically, Disc gel electrophoresis, gel fiteration and amino acid analyzer were used. Following results were obtained; 1. Nitrogen solubility of the soybean meal in $Na_{2}SO_{4},\;MgSO_{4},\;Na_{2}CO_{3},\;NaCl\;and\;Na_{2}HPO_{4}$ solutions of various concetrations were determinated. The salt soluble protein of soybean meal was highly dispersible on 0.4M $Na_{2}SO_{4}$ solution and the extractability of protein was 33%. 2. From the quantitative fractionation of soybean proteins, albumin content (46.0%) was highest followed by globulin (33.9%), glutelin (19.5%) and prolamin (2.4%). During Meju incubation period, albumin and prolamin increased gradually but glutelin decreased. Globulin content was not changed substantially. 3. When albumin was fractionated by Sephadex G-200, the following results were obtained. Soybean albumin showed fraction which was reduced to 3 fraction at 0-day of incubation. The number of fraction, however increased to 8 after 6-day of incubation. 4. Amino acids of albumin in soybean and Meju appeared to be 17 kinds. Glutamic acid and aspartic acid were the highest. In amino acid composition of cooked soybean albumin, arginine, aspartic acid, glutamic acid and glycine remained higher than those of Meju throughout incubation period. 5. The major fraction of albumins from soybean and Meju fractionated by Sephadex G-200 showed 17 kinds of amino acid. Aspartic acid and glutamic acid were the highest. During Meju incubation period, the change of amino acid composition was investigated; threonine, serine, lysine, histidine, alanine, isoleucine, leucine, phenylalanine and $NH_3$ was increased gradually, the others decreased. 6. According to the electrophoretic pattern, soybean protein showed 13 bands which decreased to 3-after cooking. During incubation, those bands increased gradually to 10 bands after 6-days.
This study was carried out to acguire some basic information on nutritional and lhysiological effects of vesicular arbuscular mycorrhizae(VAM) on mulberry trees inoculated with Glomus mosseae, Gerd. & Trap. grown in clay for 65 days and treated with 5 different levels of phosphorus, ie 30, 60, 120, 240, 480 ppm as (NH4)2 HPO4. At the End of the expermental period the levels of fixed phosphate in the soil was measared. And the native VAM fungi were collected to select the most effective VAM species on mulberry tree. The nutritional and biochemical effects of VA mycorrhizae on the mulberry leaves were also studied. Those results are as follow. 1. The mulberry trees grown in clay and inoculated with VAM were heavier in shoot dry weight as much as 197% than uninoculated plants. But in vermioulite, uninoculated mulberry trees were heavier as much as 135% than inoculated. 2. The rates of endo mycorrhizal formation in clay was highest at 60 ppm level of phosphorus, and vesicles in roots were formed in 240ppm and 480ppm level of phosphorus, but not in 30ppm, 60ppm and 120ppm. 3. The greatest growth responses of VAM inoculation was found at 60ppm level of hosphorus, and the optimum phosphorus level for VAM responese appeared to be 60ppm. 4. VAM was also to absorb soil-fixed phosphate. VAM abosrbed Fe-bound phosphate most efficiently and Ca-bound phosphate with ease but not Al-bound phosphate and Al-Fe occluded phosphate. 5. Three species of Gigaspora and one species of Glamus growing naturally in mulberry plantations were collected and tested for the growth responses. Gigaspora tricalipta and Gigaspora calospora revealed the greatest growth responses on mulberry tree among tested VAM fungi. 6. Mulberry leaves inoculated with VA mycorrhizal fungi contained 9.8% more phosphate and 15.2% more nitrogen, 22.2% more water-soluble carbohydrates and 15.2% more proteins than uninoculated plants. 7. The electrophoretic pattern of mulberry leaf protein inoculated with VAM fungi has 19 bands. 8. The patterns of peroxidase zymogram and Amulase zymogram were different between the mulberry leaves inoculated and uninoculated with VA mycorrhizal fungi. The peroxidase zymogram of inoculated leaves has 1 less majour band than unioculated leaves The amylase zymogram of inoculated leaves has 2 bands near the +pole, but that of uninoculated leaves has 1 band near the $pole.
Background : The NF-${\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.
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.
Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.
The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.
Lee, Jieun;Choi, Eun-Ji;Park, Sun Young;Jeon, Ga Young;Jang, Ja-Young;Oh, Young Jun;Lim, Seul Ki;Kim, Tae-Woon;Lee, Jong-Hee;Park, Hae Woong;Kim, Hyun Ju;Jeon, Jung Tae;Choi, Hak-Jong
Microbiology and Biotechnology Letters
/
v.42
no.3
/
pp.267-274
/
2014
High pressure processing (HPP) is a non-thermal method used to prevent bacterial growth in the food industry. Currently, pasteurization is the most common method in use for most milk processing, but this has the disadvantage that it leads to changes in the milk's nutritional and chemical properties. Therefore, the effects of HPP treatment on the microbiological and chemical properties of milk were investigated in this study. With the treatment of HPP at 600 MPa and $15^{\circ}C$ for 3 min, the quantity of microorganisms and lactic acid bacteria were reduced to the level of 2-3 log CFU/ml, and coliforms were not detected during a storage period of 15 d at $4^{\circ}C$. An analysis of milk proteins, such as ${\alpha}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin by on-chip electorophoresis revealed that the electrophoretic pattern of the proteins from HPP-treated milk was different from that of conventionally treated commercial milk. While the quantities of vitamins and minerals in HPP-treated milk were seen to be comparable to amounts found in raw milk, the enzyme activity of lipase, protease and alkaline phosphatase after HPP treatment was reduced. These results suggest that HPP treatment is a viable method for the control of undesirable microorganisms in milk, allowing for minimal nutritional and chemical changes in the milk during the process.
Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.
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