Climacteric phenomon in fruits was delayed ten days on the sub-atmospheric treatment than the normal atmospheric one. Average lipid content in tomatoes sampled was composed of neutral lipid (49%), phospholipid(31%) and glycolipid (13%). Seven constituents each from neutral and phospholipid were seperated through thin-lyaer chromatography. Changes on lipid content during treatments were associated with the climacteric pattern and the advance of maturity; At the climacteric on-set, the fruit color was light pink, and the content of neutral lipid was reduced to minimum. After this stage, this content was reversely increased. The maturity was related with diglyceride and sterol ester in neutral lipid, and phosphatidyl choline and phosphatidyl ethanolamine in phospholipid. The changes of two constituents each on both neutral and phospholipid were reversed each other. It was concluded that changes of lipid content in tomato fruits were closely associated with the advance of fruit maturity and climacteric pattern.
To study lipid components of Panax ginseng produced in Korea, the lipids of fresh ginsengs were extracted with the mixture of chloroform-methanol (2:1, v/v) and those of dried ginsengs were extracted with diethyl ether respectively. The lipid components extracted were separated and quantitated by column, thin layer and gas-liquid chromatographies. The results were summarized as follows : 1. Fresh ginseng contained 0.62% total lipid of which 45.28% were neutral lipids, 18.12% glycolipids, and 36.60% phospholipids. But dried ginseng contained 0.89% total lipids of which 86.48% were neutral lipids, 9.20% glycolipids, and 4.32% phospholipids. 2. Triglycerides (37.6 to 42.5% of the total neutral lipids) and sterol esters (16.5 to 19.6%) in all the fresh and dried ginseng were the major components among the neutral lipids. Monoglycerides, diglycerides, free fatty acids and free sterols were minor components. 3. Digalactosyl diglycerides (23.5% of the total glycolipids) in the fresh ginseng and steryl liglycosides (28.9%) in the dried ginseng were predominant components among the glycopids, respectively, Esterified steryl glycosides and monogalactosyl diglycerides were also identified, and four unknown spots in the fresh ginseng and two unknown spots in the dried ginseng were present. 4. Phosphatidyl cholines (31.3 to 31.9% of the total phospholipids) and phosphatidyl glycerols (34.8 to 36.7%) in all the fresh and dried ginseng were the major components among the phospholipids. Phosphatidyl inositols and phosphatidyl ethanolamines were also identified. 5. The major fatty acids in the fresh and dried ginseng were linoleic $(62.29{\sim}64.32%)$, palmitic $(13.16{\sim}15.63%)$, oleic $(5.73{sim}7.23%)$ and linolenic $(5.73{sim}7.23%)$. The fatty acid compositions in neutral lipid fraction was similar to the pattern in those of the total lipids. But glycolipid and phospholipid fractions contained a lower percent of linoleic acid and a higher percent of palmitic acid than the neutral lipid fraction.
Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
Journal of Life Science
/
v.21
no.7
/
pp.1032-1038
/
2011
Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.
Protective effects of natural components including genistein (4',5,7-trihydroxyisoflavone) from Glycine max MERRILL on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. Genistein $(10{\sim}100\;{\mu}m)$ suppressed photohemolysis in a concentration-dependent manner, and was more effective than the lipid peroxidation chain blocker, ${\alpha}$-tocopherol (Vit. E). Glycoside of genistein, genistin, the water-soluble antioxidant, L-ascorbate, and the iron chelator, myo-inositol hexaphosphoric acid dodecasodium salt (sodium phytate) did not exhibit protective effect against photohemolysis. L-Ascorbate and sodium phytate stimulated photohemolysis at high concentration $(500\;{\mu}m)$. ${\alpha}$-Carotene 3,3'-diol (lutein), a singlet oxygen $(^1O_2)$ quencher, exhibited pronounced protective effect, an indication that $^1O_2$ is important in photohemolysis sensitized by rose-bengal. Reactive oxygen scavenging activities $(OSC_{50})$ of natural antioxidants including genistein on reactive oxygen species (ROS) generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were in the order of sodium phytate > L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. $OSC_{50}$ value of genistein, genistin, ${\alpha}$-tocopherol, L-ascorbate, and sodium phytate were 41.0, 109.0, 9.0, 5.2, and $0.56{\mu}m$ respectively. The order of free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ was L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. These results indicate that genistein can function as an antioxidant in biological systems, particularly skin exposed to solar UV radiation by scavenging $^1O_2$ and other ROS, and to protect cellular membranes against ROS.
Kim, Cho-Hee;Kim, Min-Young;Lee, Su-Yeon;Moon, Ji-Young;Han, Song-Iy;Park, Hye-Gyeong;Kang, Ho-Sung
Journal of Life Science
/
v.19
no.8
/
pp.1073-1080
/
2009
A number of studies have demonstrated that the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce the risks of colorectal, oesophageal and lung cancers. NSAIDs have been shown to exert their anti-cancer effects through inducing apoptosis in cancer cells. The susceptibility of tumor cells to anti-tumor drug-induced apoptosis appears to depend on the balance between pro-apoptotic and anti-apoptotic programs such as nuclear factor kB (NF-kB), phosphatidylinositol 3-kinase (PI3K)-Akt/protein kinase B (PKB) and MEK1/2-ERK1/2 pathways. We examined the effects of pro-survival PI3K and ERK1/2 signal pathways on cell cycle arrest and apoptosis in response to NSAIDs including sulindac sulfide and NS398. We show that simultaneous inhibition of the Akt/PKB and ERK1/2 signal cascades could synergistically enhance the potential pro-apoptotic activities of sulindac sulfide and NS398. Similar enhancement was observed in cells treated with sulindac sulfide or NS398 and 100 ${\mu}$M genistein, an inhibitor of receptor tyrosine kinases (RTKs) that are upstream of PI3K and MEK1/2 signaling. We further demonstrate that NAG-1 is induced and plays a critical role(s) in apoptosis by NSAIDs-based combined treatment. In sum, our results show that combinatorialtreatment of sulindac sulfide or NS398 and genistein results in a highlysynergistic induction of apoptotic cell death to increase the chemopreventive effects of the NSAIDs, sulindac sulfide and NS398.
Background: Phospholipase C(PLC) plays a central role in cellular signal transduction and is important in cellular growth, differentiation and transformation. There are currently ten known mammalian isozymes of PLC reported to this date. Hydrolysis of phosphatidylinositol 4,5-bisphosphate($PIP_2$) by PLC produces two important second messengers, inositol 1,4,5-trisphosphate($IP_3$) and diacylglycerol. PLC-${\gamma}1$, previously, was known to be activated mainly through growth factor receptor tyrosine kinase. Other mechanisms of activating PLC-yl have been reported such as activation through tau protein in the presence of arachidonic acid in bovine brain and activation by $IP_3$, phosphatidic acid, etc. Very recently, another PLC-${\gamma}1$ activator protein such as tau has been found in bovine lung tissue, which now is considered to be AHNAK protein. But there has been no report concerning AHNAK and its associated disease to this date. In this study, we examined the expression of the PLC-${\gamma}1$ activator, AHNAK, in lung cancer specimens and their paired normal. Methods: From surgically resected human lung cancer tissues taken from twenty-eight patients and their paired normal counterparts, we evaluated expression level of AHNAK protein using immunoblot analysis of total tissue extract Immunohistochemical stain was performed with primary antibody against AHNAK protein. Results: Twenty-two among twenty-eight lung cancer tissues showed overexpression of AHNAK protein (eight of fourteen squamous cell lung cancers, all of fourteen adenocarcinomas). The resulting bands were multiple ranging from 70 to 200 kDa in molecular weight and each band was indistinct and formed a smear, reflecting mobility shift mainly due to proteolysis during extraction process. On immunohistochemistry, lung cancer tissues showed a very heavy, dense staining with anti-AHNAK protein antibody as compared to the surrounding normal lung tissue, coresponding well with the results of the western blot Conclusion: The overexpression of PLC-${\gamma}1$ activator protein, AHNAK in lung cancer may provide evidence that the AHNAK protein and PLC-${\gamma}1$ act in concerted manner in carcinogenesis.
Jin, Kyong-Suk;Park, Jung Ae;Lee, Ji Young;Kang, Ji Sook;Kwon, Hyun Ju;Kim, Byung Woo
Microbiology and Biotechnology Letters
/
v.42
no.2
/
pp.170-176
/
2014
In this study, the anti-oxidative and anti-inflammatory activities of Euptelea pleiosperma ethanol extract (EPEE) were evaluated using in vitro assays and cell culture model systems. EPEE possessed a more potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl than the ascorbic acid used as a positive control. EPEE effectively suppressed lipopolysaccharide (LPS), in addition to hydrogen peroxide induced reactive oxygen species on RAW 264.7 cells. Furthermore, EPEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1) and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), dose and time dependently. The modulation of HO-1 and Nrf2 expression might be regulated by mitogen-activated protein kinases and phosphatidyl inositol 3 kinase/Akt as their upstream signaling pathways. On the other hand, EPEE inhibited LPS induced nitric oxide (NO) formation without cytotoxicity. Suppression of NO formation was the result of the down regulation of inducible NO synthase (iNOS) by EPEE. Suppression of NO and iNOS by EPEE may be modulated by their upstream transcription factor, nuclear factor ${\kappa}B$, and AP-1 pathways. Taken together, these results provide important new insights into E. pleiosperma, namely that it possesses anti-oxidative and anti-inflammatory activities, indicating that it could be utilized as a promising material in the field of nutraceuticals.
The Journal of the Korean Society for Microbiology
/
v.8
no.1
/
pp.7-11
/
1973
The authors identified eighty-eight Shigella cultures among about four thousands specimens collected from all over the country in 1972. Of eighty-eight cultures, seventy-seven cultures belonged to Shigella flexneri and eleven cultures to Shigella sonnei. None of cultures belonging to either subgroup A or C was detected in 1972. Of seventy-seven cultures of Shigella flexneri one was $B_{1b}$, fifty-six were $B_{2a}$, nine were $B_{3a}$, six were $B_{4a}$, three were By and one was each of $B_{3b}$ and $B_{3c}$. Of eleven cultures of Shigella sonnei seven cultures appeared to be phase I and the others phase II. Although there was quite a difference found in the incidence of isolating Shigella organisms between different areas as shown in Table 1, it would not be possible to understand that there might not have been the cases or carriers of Shigella in the areas where the organisms were not isolated in 1972. Concerning the biochemical properties it was not possible to compare the results obtained from the decarboxylase and dihydrolase tests with them obtained in previous years except that of lysine decarboxylase tests since they were not reported individually by the different serotypes in the previous reports. These results obtained in 1972 would be the data for the future comparison. In regards to the antibiotics-sensitivity of Shigella cultures the most of them showed sensitive results to nitrofurantoin, ampicillin, cephalosporin, gentamycin and geopen, and the majority of them appeared to be resistant to cloxacillin, tetracycline and streptomycin by means of the In Vitro tests.
For investigation of new utilization as jang-products, Meju was prepared using barely bran. As barley meju was fermented, change of pH was $5.2{\sim}5.6$, it was indistinguishable change. L-value of color was changed from 46.9 to 60.3, that meant it was getting moe dark. The counts of aerobic bacteria were $4.8{\times}10^7{\sim}5.6{\times}10^9$ CFU/g, it was extraordinarily increased during fermentation. Counts of Yeast, molds, and bacteria were $9.1{\times}10^6{\sim}5.0{\times}10^8$ CFU/g, $8.3{\times}10^5{\sim}6.9{\times}10^7$, and $2.0{\times}10^2{\sim}4.5{\times}10^6$ CFU/g, respectively. Crude ash content was $3146.0{\sim}7147.4$ mg%. The level of K was the highest in quantity among the crude ash in barely meju. 7 free sugars(i.e., raffnose, stachyose, inositol, fructose, glucose, arabinose, and maltose), 3 volatile organic acid(i.e., acetic acid, propionic acid, and butyric acid) and 4 non-volatile organic acid(i.e., fumaric acid, ${\alpha}-ketoglutaric$ acid, malic acid, and citric acid) were detected. The content of free amino acid was $596.3{\sim}1580.8$ mg%. Glutamic acid was most abundant component among the amino acids, 2nd abundant component was alanine, it's content was $79.9{\sim}165.3$ mg%, 3rd abundant component was leucine, it's count was $41.7{\sim}161.6$ mg%. Finally, essential amino acid content was revealed $33.2{\sim}40.38%$.
The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$$m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$$m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$$m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$$m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$$d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.
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