Changes in lipid components of Gae-bul, Urechis unicinctus, during hot-air drying ($40^{\circ}C$, 7 hrs) were studied. Raw sample contained 1.3% total lipid (TL) which consisted of 35.1% neutral lipid (NL), 18.0% glycolipid (GL) and 46.9% phospholipid (PL), and dried sample contained 5.3% TL which consisted of 51.8% NL, 20.5%GL and 27.7% PL. There were about 40% decrease in PL content and a slight increase in NL content during drying. The NL of raw sample mainly consists of triglyceride (TG, 39.8%), free sterol (FS, 39.6%), free fatty acid (FFA, 12.2%). and also identified diglyceride (DG), monoglyceride and esterified sterol and hydrocarbon in less quantify. The percent of TG and FS decreased, while that of FFA and DG increased during drying. And main components in the PL were phosphatidyl choline (PC,45.6%), phosphatidyl ethanolamine (PE,34.8%), and followed by phosphatidyl serine (PS) and an unknown substance. In the components of PL, PE, PS and PC decreased slightly in order during drying. And major fatty acids of raw and dried samples were generally 16:0, 18: 1, 18:3, and 20:5. The content of the polyenoic acid such as 20:5 decreased. while the saturated acid increased slightly during drying.
Voltage-sensitive ion channels contribute to establishment of the cell excitablity and the generation of the cellular function. At hamster oocytes in the primitive stage during developing process, an inward current elicited by voltage pulses was found to be carried mainly by $Ca^{2+}$. Even at present, $Ca^{2+}$ channels serve as the most probable route to pass this inward current but there is no evidence of the presence of this channels in eggs. To date, both the characteristic properties and the physiological role in the early stage of development remain unclear. Here we examined the characteristic properties of the inward current and changes in this currents at unfertilized oocytes, fertilized zygotes and two-cell embryos using whole-cell voltage clamp technique. The inward current carried reportedly by $Ca^{2+}$ was remained following removing external $Ca^{2+}$ but completely abolished by further replacement of impermeants such as tetramethylammonium ion ($TMA^+$) or $choline^+$ instead of $[Na^+]_0$. Tetrodotoxin did not affect on this inward current remained at $[Ca^{2+}]_0$-free condition. Removal of $Na^+$ ion out of the experimental solution clearly decreased the current. After adding 2mM $Ca^{2+}$ to the $Na^+$-free media, the inward current was restored. Interestingly, this current carried by either $Ca^{2+}$ or $Na^+$ was decreased by the reduction of intracellular $Cl^-$ concentration, or by $Cl^-$ channel blockers such as niflumic acid, DIDS and SITS. When $Cl^-$ concentration was lowered without changes in other ionic components, this inward current was reduced. At fertilized oocytes and two-cell embryos, the inward current carried by $Ca^{2+}$ and $Na^+$ was severely reduced. Also $Cl^-$ component could not be observed. From these results, the inward current is composed of $Ca^{2+}$, $Na^+$ and $Cl^-$ component, suggesting that the channel carrying this inward current is not selective specifically to $Ca^{2+}$. During early stage of development, the voltage-sensitive ion current seems not to contribute essentially to the cell cleavage and differentiation. The loss of $Cl^-$ component after fertilization suggests that $Cl^-$ may play a role in maintaining the viability of unfertilized ova.
Phospholipase D (PLD) was immobilized on a submicro-porous membrane through covalent immobilization. The immobilization was conducted on the porous membrane surface with the treatment of polyethyleneimine, glutaraldehyde, and the anhydrase, in sequence. The immobilization was confirmed using X-ray photon spectrometer. The pH values of phosphatidylcholine (PC) dispersion solution with buffer were monitored with respect to time to calculate the catalytic activities of PC for free and immobilized PLD. The catalytic rate constant values for free PLD, immobilized PLD on polystyrene nanoparticles, and immobilized PLD on a porous cellulose acetate membrane were 0.75, 0.64, and 0.52 s-1, respectively. Reusability was studied up to 10 cycles of PC hydrolysis. The activity for the PLD immobilized on the membrane was kept to 95% after 10 cycles, and comparable to the PLD on the nanoparticles. The stabilities for heat and storage were also investigated for the three cases. The results suggested that the PLD immobilized on the membrane had the least loss rate of the activity compared to the others. From these studies, the porous membrane was feasible as a carrier for the PLD immobilization in the production of phosphatidic acid.
Total lipid contents in rice bran for Poong-San(Tongil) and Dong-Jin(Japonica) were 16.13% and 16.97%, respectively. Neutral lipids for Pong-San(75.20%) were slightly higher than those for Dong-Jin(73.69%), whereas contents of glycolipid for Poong-San(16.71%) were lower than those for Dong-Jin(22.80%). Contents of phospholipid in Poong-San(8.09%) were much higher than those in Dong-Jin(3.51%). Acid, peroxide and thiobarbituric acid value of total lipids extracted from rice bran of Poong-San were slightly lower than those of Dong-Jin. Iodine value showed the reverse trend. The neutral lipids were fractionated and identified as hydrocarbon, esterified sterol, triglyceride, free fatty acid, free sterol, diglyceride and monoglyceride. Triglyceride contents were less than common edible oils, but diglyceride and monoglyceride contents were higher. Among the glycolipids contained in the polar lipids, esterified sterylglycoside(11.46%) was the most abundant. Of the phospholipids, phosphatidyl ethanolamine, phosphatidyl inositol and phosphatidyl choline were the major components. Main fatty acids in the total lipids, three lipid components and stepwise eluted individual lipids were olelic acid, linoleic acid and palmitic acid. The fatty acid composition of the neutral lipids was similar to that of the total lipids. In glycolipids, the content of linoleic acid was higher than that of oleic acid, and palmitic acid was predominant in the fatty acid composition of the esterified sterylglycoside.
The studies are conducted on the changes of soybean lipids in terms of physicochemical characteristics, lipid classess and fatty acid composition during the fermentation process of soybean Koji preparation for daenjang (soybean paste) in a model system using cooked soybean inoculated by Aspergillus oryzae. The total lipids contents were increased during soybean Koji preparation, generally iodine values decreased but acid values increased. Total lipids of soybean Koji consisted of about 90.6% neutral lipids, 7.6% phospholipids and 1.8%, glycolipids indicating that phospholipids contents of soybean Koji was increased when compared to those of cooked soybean. The major components of nonpolar lipids in soybean Koji were free fatty acids(39.6%) and triglycerids(29.2%). Free fatty acids increased as the triglycerides decreased during soybean Koji preparation by the hydrolysis of lipase action. The major components of polar lipids in soybean Koji were phosphatidyl choline and phosphatidyl ethanolamine. Differences were observed in the composition of the polar lipids of cooked soybean and soybean Koji. A little changes also occurred in fatty acid compositions of total lipids, triglycerides and free fatty acids fractions in soybean Koji preparation. Especially a considerable increase of linoleic acid in free fatty acid fraction was observed in soybean Koji.
LEE Eung-Ho;WADA Shun;KOIZUMI Chiaki;OHSHIMA Toshiaki;NONAKA Junsaku
Korean Journal of Fisheries and Aquatic Sciences
/
v.17
no.4
/
pp.291-298
/
1984
The extracted hagfish (Eptatretus burgeri) flesh lipid was separated into following fractions by column chromatography on Bio-beads SX-2 and Sephadex LH-20 prior to gab chromatographic analysis of their fatty acid compositions: polar lipid, triglyceride and free fatty acid. The major fatty acids of total lipid and triglyceride in hagfish were $C_{16:0},\;C_{16:1},\;and\;C_{18:1}$. The ratio of $C_{18:0}/C_{18:1}$ in the total lipid and triglyceride of hagfish was 0.1. The polar lipid of the hagfish muscle was mainly composed of phosphatidyl choline ($65.5\%$) and phosphatidyl ethanolamine ($28.0\%$). The triglyceride obtained was fractionated into four fractions by HPLC on the basis of partition numbers. Both the fatty acid composition and triglyceride composition on the basis of the total carbon number in the acyl chains of the triglyceride were analysed by the GLC. From the information obtained on triglyceride compositions based on the total carbon number by GLC and the partition number by HPLC and fatty acid composition by GLC, the combination of fatty acid in each triglycerides was estimated. A computer was used for estimation of the fatty acid combination in the triglyceride because hagfish lipid triglyceride was composed of various kinds of fatty acids. Fortyfour kinds of triglyceride were estimated. The major triglycerides in hagfish flesh lipid were found to those of ($1{\times}C_{16:0},\;2{\times}C_{18:1};\;13.5\%$), ($1{\times}C_{16:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;7.2\%$), ($1{\times}C_{16:1},\;2{\times}C_{18:1};\;5.4\%$), ($2{\times}C_{16:0},\;1{\times}C_{22:5};\;5.2\%$), ($1{\times}C_{14:0},\;2{\times}C_{18:1};\;4.5\%$), ($2{\times}C_{18:1},\;1{\times}C_{22:5};\;3.6\%$), ($1{\times}C_{14:0},\;1{\times}C_{18:0},\;1{\times}C_{18:1};\;2.7\%$) and ($1{\times}C_{14:0},\;1{\times}C_{16:0},\;1{\times}C_{18:2};\;2.2\%$).
Quantitative analysis of MR spectrum depending on mole concentration of the contrast media in cereberal metabolite phantom was performed. PRESS pulse sequence was used to obtain MR spectrum at 3.0T MRI system (Archieva, Philips Healthcare, Best, Netherland), and the phantom contains brain metabolites such as N-Acetyl Asparatate (NAA), Choline (Cho), Creatine (Cr) and Lactate (Lac). In this study, optimization of MRS PRESS pulse sequency depending on the concentration of contrast media (0, 0.1 and $0.3mmol/{\ell}$) was evaluated for various repetition time(TR; 1500, 1700 and 2000 ms). In control (cotrast-media-free) group, NAA and Cho signals were the highest at TR 2000 ms than at 1700 and 1500 ms. Cr had the highest peak signal at TR 1500 ms. When concentration of contrast media was $0.1mmol/{\ell}$, the metabolites were increased NAA 73%, Cho 249%, Cr 37% at TR 1700 ms compared with other TR, and also signal increased at $0.3mmol/{\ell}$, In $0.5mmol/{\ell}$ of contrast agent, cerebral metabolite peaks reduced, especially when TR 1500 ms and 2000 ms they decreased below those of control group. The ratio of metabolite peaks such as NAA/Cr and Cho/Cr decreased as the concentration of the contrast agent increased from 0.1 to $0.5mmol/{\ell}$. Authors found that the optimization of PRESS sequence for 0.3T MRS was as follows: low density of contrast agent ($0.1mmol/{\ell}$ and $0.3mmol/{\ell}$) made the highest signal intensity, while high density of contrast agent reveals the least reduction of signal intensity at 1700 ms. In conclusion, authors believe that it is helpful to reduce TR for acquiring maximum signal intensity.
The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes ${\beta}$, ${\gamma}$ and ${\delta}$ were obtained and the characteristics of PLC activity on various concentrations of free $Ca^{2+}$ were observed. The activity of PLC was increased with increasing $Ca^{2+}$ concentration and the activity PLC ${\delta}$ was increased higher in the presence of phosphatidyl choline(PC) than in the abscence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate($PIP_2$) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the $PIP_2$ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2%-1% cholic acid. The activities of PLC isoyzmes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC ${\beta}$and ${\delta}$. A large amount of PLC ${\delta}$ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11%-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in $Go{\alpha}$ & Gmix, and the activities of PLC ${\beta}$ and ${\delta}$ were increased in $G{\beta}{\gamma}$ and $Gi{\alpha}$. Activities of PLC ${\beta}$ and ${\gamma}$ were decreased in $Gt{\alpha}$ but PLC ${\delta}$ increased.
Purpose : To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. Materials and Methods : All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% $D_2O$. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), lutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. Results : Symmetrical 2D-COSY and 2D-NOESY pectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,$H{\alpha}$)) at 2.50ppm and methylene protons ($CH_2$,(3,$H_B$)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons ($CH_3$) were observed in NAA. Cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons ($CH_2$(3)) at 2.11 ppm and methylene protons ($CH_2$(4)) at 2.35 ppm, and between the methylene protons($CH_2$ (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. Conclusion : The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.
The purpose of the present study is to investigate the effects of green tea catechin on microsomal phospholipase $A_2(PLA_2)$ activity and the arachidonic acid (AA) cascade in hearts of microwave exposed rats. Sprague-Dawley male rats weighing $100{\pm}10$ g were randomly assigned to one normal group and three microwave exposed groups. The microwave exposed groups were subdivided into three groups: catechin free diet (MW) group, 0.25% catechin (MW-0.25C group and 0.5% catechin (MW-0.5C) group according to the levels of dietary catechin supplementation. Rats were sacrificed $6^{th}$ day after microwave irradiations (2.45 GHz, 15 min). The heart microsome $PLA_2$ activity in the MW group was 130% greater than that of normal groups, whereas there was no significant difference between normal group and MW-0.25C, MW-0.5C group. The per- centage phosphatidyl ethanolamine (PE) hydrolyzed in the heart microsome in the MW was increased 54% by microwave irra- diation, whereas there was no significant difference between normal group and MW-0.5C group. The percentage phosphatidyl choline (PC) hydrolyzed in the heart microsome in the MW group was increased by 104% and by microwave irradiation, whereas there was no significant difference between normal group and MW-0.5C group. The formation of thromboxane $A_2(TXA_2)$ in the heart microsome was 70% greater in the MW group than in the normal group. However, the MW-0.25C and MW-0.5C group maintained the normal level. The formation of prostacyclin ($PGI_2$) in the heart microsome was 21% lower in the MW group than in the normal group, while that of MW-0.25C and MW-0.5C group were maintained in the normal group. The heart microsomal thiobarbituric acid reactive substances (TBARS) concentrations, as an index of lipid peroxide, were 71% greater in the MW group, as compared with normal group. However, the MW-0.25C and MW-0.5C group were 4.6% and 9.2% lower, respectively, than that of MW group. In conclusion, heart function appeared to be improved by green tea catechin supplementation due to its antithrombus action, which in return controls the AA cascade system.
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