A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in bovine, porcine, chicken, flatfish and japanese eel muscle has been developed and validated. The separation condition for HPLC/UV was optimized with phenyl hexyl ($4.6{\times}150mm$, $5{\mu}m$) column with 10 mM monobasic sodium phosphate buffer (pH 2.5)/acetonitrile (50/50, v/v) as the mobile phase at a flow rate of 1.0 mL/min and detection wavelength was set at 254 nm. Residues were extracted from tissue by blending with methanol and lipid materials were removed with n-hexane. Then, the methanol extract was evaporated to dryness under a nitrogen stream, reconstituted in the mobile phase. Aliquot of the organic extract was decanted and filtered through $0.45{\mu}m$ syringe filter. The $20{\mu}L$ of the resulting solution was injected into the HPLC system. The calibration ranges were $0.5{\sim}5{\mu}g/g$ and calibration curves were linear with coefficients of correlation better than 0.95. The limits of quantification were $0.5{\mu}g/g$ for all muscles. The recoveries of bovine, porcine, chicken, flatfish and japaneseel muscles were 99.8%, 102.4%, 91.0%, 104.0% and 93.0%, respectively. The procedures were validated according to the CODEX guideline, determining specificity, linearity, accuracy, precision, quantitation limit and recovery.
Changes of the activities of the hepatic cells of female mud skipper, Boleophthalmus pectinirostris were investigated under transmission electron microscopy. Monthly changes of gonadosomatic index(GSI) and hepatosomatic index(HSI), variations of protein and nucleic acid contents(total RNA and DNA) of the liver tissues with the gonadal development phase were also studied. GSI began to increase from May(the growing stage), reaching the maximum value in late June(the mature stage), and then it began to decrease from late July(the degenerative stage), reaching the lowest value in late September. Monthly variations of HSI were negatively related to GSI. HSI decreased in the summer season when the ovary was getting mature and reached the maximum in mid October when the ovary was degenerating. In June(the mature stage), the female hepatic cells of the liver tissues became large and nuclei were hypertrophic. The amounts of glycogen particles and lipid droplets in the cells gradually decreased, while a number of granular endoplasmic reticulum increased. It was assumed that well-developed granular endoplasmic reticulum binding ribosomes are supposed to play the leading role in protein synthesis and deposition for vitellogenin in the cytoplasm. In July(the spawning period), glycogen particles and lipid droplets gradually increased, and then these substances were still observed in large quantity in August(the degenerative stage). The protein contents of the liver tissues with the gonadal phases of the ovaries were shown the maximum value($4.720{\pm}0.103\;mg/g$) in June, and afterwards gradually decreased being the minimum($3.640{\pm}0.130\;mg/g$) in July, and then gradually increased in August. The mean total RNA contents per gram of the liver tissues appeared the maximum($0.523{\pm}0.040\;mg/g$) in June, and afterwards gradually decreased to the minimum($0.158{\pm}0.006\;mg/g$) in July and slightly increased in August again. From these results, it could be assumed that protein contents were closely related to RNA contents. The mean total DNA contents per weight (gr) of the liver tissues appeared to be similar although there were some monthly fluctuations. The ratio of the mean total RNA/DNA were 0.745 in June, 0.262 in July, 0.341 in August respectively.
This study was performed to reveal nutritional excellency of popular Korean dishes, which were intended to distribute as a take-out type food suited for Americans. Four Korean dishes were selected due to their high preference from American sensory panels: Jeyuk-bokkeum (B), Kimchi-bokkeum (C), Dubu-jorim (D) and Namul (E). Nine weeks of the experimental periods were composed of two parts. In phase 1(5 weeks of the first half), high fat diets were fed to all treatments including the control group (A). In phase 2 (4 weeks of the last half), freeze-dried four Korean dishes were fed to the treatments except for the control. There was no significant difference in the final body weights among the groups as well as daily body weight gains and FER. Organ weights of the control group tended to be smaller than those of the treatment groups. For visceral fats, perirenal fat pad (RFP) and epidydimal fat pad (EFP) of the groups C and D were decreased by 20% compared with those of the group A. Total cholesterol level of the group C (94.13 mg/dL) was significantly lower than any other groups (p<0.05), but there was no significant difference in triglyceride(TG) level. HDL-cholesterol level of group B(20.38 mg/dL) was significantly higher than that of group D (17.25 mg/dL), but no significant differences were seen in groups C and E (p<0.05). In LDL-cholesterol level, the highest level was seen in group D, the lowest level in group E (p<0.05). From the findings, Kimchi-bokkeum, Dubu-jorim and Namul were superior to those of the control and jeyukbokkeum in terms of the blood lipid profiles. It might be a good Korean style take-out items suited for the Americans’ taste along with a good nutritional quality in the near future.
Sophora tonkinensis Gapnep has been used as a traditional herbal medicine in oriental regions since ancient times. In this study, the effect and mechanism of the MeOH extract of Sophora tonkinensis Gapnep (STME) on adipocite differentiation and adipogenesis in 3T3-L1 preadipocites were investigated. Treatment with STME in the concentration range of 0-200 ${\mu}g$/ml significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner, as determined by a decrease in intracellular lipid droplets and lipid contents measured by Oil Red O staining. In association with the inhibitory effect of lipid accumulation, the expressions of the proteins concerned with adipogenesis in 3T3-L1 preadipocites were also investigated. Treatment with STME reduced the expressions of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ and ${\beta}$ (C/EBP${\alpha}$ and C/EBP${\beta}$) and sterol regulatory element binding protein (SREBP), which are adipocyte specific markers. In flow cytometry analysis, the inhibitory effect of differentiation was caused by G1 arrest and following mitotic clonal expansion cease. Therefore, we also investigated the alteration of G1 phase arrest-related proteins. As a result, the expression of p21 protein was significantly increased, while the expressions of Cdk2, E2F-1 and phospho-Rb were reduced in a dose-dependent manner in STME treated 3T3-L1 cells. According to these results, STME might inhibit differentiation through G1 arrest in 3T3-L1 preadipocytes adipogenesis, and further studies, which are in progress, have to be completed to identify the active compounds.
The present study was performed to identify the factors influencing on early pregnacy and embryo implantation in rabbit. Serum, uterine fluid, and uterine tissue were collected on day 0, 3, 5, 7 and 9 of pregnancy. The intrauterine environment of receptive phase and refractory phase was compared by measuring protein synthetic capacity of endometrium, amino acid composition and concentrations of lipids(phospholipid, cholesterol). The results obstained were as follows : 1. The concentrations of total protein were significantly increased (p<0.01) on day 5 ($7.00{\pm}0.55$), 7($6.29{\pm}0.65$), and 9($6.34{\pm}0.61$), compared to those on day 0($5.50{\pm}0.12g/100m{\ell}$) in serum. The concentration of albumin on day 0 was $0.81{\pm}0.05$ and reached maximum on day 5 ($1.59{\pm}0.07g/100m{\ell}$) in serum. The concentrations of total protein were significantly increased(p<0.01) on day 5($1.56{\pm}0.10$), 7($1.99{\pm}0.22$), compared to those on day 0($0.38{\pm}0.02g/100m{\ell}$) in uterine fluid. The concentration of albumin on day 5($0.78{\pm}0.05g/100m{\ell}$) was higher than those on the other days in uterine fluid. 2. The incorporation rates of [$^3H$]-leucine into protein were significantly increased(p<0.01) on day 5 ($919.6{\pm}97.5$), 7($1445.4{\pm}95.9$) and 9($450.38{\pm}28.71$), compared to those on day 0($328.2{\pm}38.9cpm/mg$ protein) in endometrium. The incorporation rates in colehicine-treated endometrium on day 5 ($1341.9{\pm}73.8$), 7($1729.4{\pm}63.3cpm/mg$ protein) were significantly higher(p<0.01) than those on the other days. 3. The compositions of amino acid were not distinctly changed during early pregnancy in serum. The composition ratios of methione, lysine were distinctly decreased on day 3, compared to those on day 0 in uterine fluid. Those of glycine, alanine were increased on day 9, compared to those on other days but his tidine decreased in uterine fluid. 4. The concentrations of total phospholipid and total cholesterol were significantly decreased(p<0.01) on day 3($77.9{\pm}15.5$, $61.5{\pm}21.2$), compared to those on day 0($164.0{\pm}33.9$, $167.2{\pm}46.2mg/100m{\ell}$)in serum. The concentrations of total phospholipid and total cholesterol on day 9 ($47.3{\pm}13.4$, $37.7{\pm}9.6mg/100m{\ell}$) were significantly higher(p<0.01) than those on the other days in uterine fluid. 5. Total phopholipid/total cholesterol ratios were not significantly changed during early pregnancy in serum. However, total phospholipid/total cholesterol ratios on day 5 ($2.00{\pm}0.42$), 7 ($1.11{\pm}0.77$) and 9 ($1.47{\pm}0.30$) were higher than those on day 3($0.84{\pm}0.41$) in uterine fluid. 6. The concentrations of phosphatidylcholine and phosphatidylserine were significantly increased (p<0.01) on the other days, compared to those on day 0 during early pregnancy in serum. The concentrations of phosphatidylcholine were significantly increased(p<0.01),compared to those on day 0 and those of phosphatidyl-ethanolamine were consistently increased but not significant in early pregnancy in uterine fluid.
In this article, we investigate into the structural changes of liposome to design its functional membranes by the synthesis of two types of liposomes, DPPC liposome and DTAB of hydrocarbon substance/DPPC liposome. The changes of membrane structures are evaluated by the CF fluorescent intensity measured above and below the phase transition temperature of the membrane, $t_c=41^{\circ}C$. CF fluorescent intensities are enhanced by the CF leakage from DPPC liposome at $45^{\circ}C$, while no changes are observed at $20^{\circ}C$. Under the same conditions, it is observed that the intensity enhanced by CF leakage from DPPC/DTAB liposome is larger than that of DPPC liposome alone, which suggests that DPPC/DTAB liposome has irregular arrangement. Under the presence of $Ca^{2+}$, Quin 2 fluorescent intensity in either DPPC liposome or DPPC/DTAB liposome is significantly increasing at $45^{\circ}C$, while almost none of the changes are observed at $20^{\circ}C$. The fluorescent intensity of DPPC liposome turns out to be larger than that of DPPC/DTAB liposome, which suggests that the DPPC/DTAB liposome is structurally more stable than the DPPC liposome. Additionally, when the analysis is done to observe changes in the shapes of membrane surfaces with ANS fluorescent, ANS fluorescent under DPPC or DPPC/DTAB liposome shows each of different appearances at $45^{\circ}C$ and $20^{\circ}C$ respectively. This result indicates that its respective membrane fluidity is changing above and below of the designated temperatures in phase transition. As to the magnitude of change of its membrane fluidity, DPPC liposome is much larger than DPPC/DTAB liposome. As far as the temperature in phase transition measured by DSC are concerned, it is $41^{\circ}C$ and $32^{\circ}C$ for DPPC and DPPC/DTAB liposome respectively, which suggests that DPPC/DTAB liposome has an irregular molecular arrangement in its structure. That is, it is summed up that DPPC/DTAB turns out to be structurally stable, even so, its structure is irregularly arranged.
This study was designed to identify the ultrastructural changes of mouse endometrium during peri-implantation period and obtain the fundamental information for the establishment of 3-dimensional culture system of mouse endometrial cells in vitro. The used female ICR mice ($6{\sim}8$ wks) were conducted on pregnant. The biopsies were obtained from whole uterus at cycle day 1 (D1) and day 5 (D5) after hCG injection and mating. The biopsies materials were fixed 2.5% glutaraldehyde and 1% osmium tetroxide. Subsequently, for observation using light and transmission electron microscopy (LM and TEM), they were dehydrated and embedded in Epon and the embedded biopsies were sectioned and stained. For scanning electron microscopy (SEM), the fixed specimens were dehydrated, dried and coated with gold. 1) For LM, the biopsied materials at D5 (late secretory phase) were appeared the extended stromal layer by increased connective tissues and the fully developed endometrial glands and vessels compared with D1 (early secretory phase). 2) For TEM, the mouse endometrium was consisted of 3-layers, a simple polarized columnar epithelial cells, basement membrane and stromal cells. At D5, the distribution of microvilli, endoplasmic reticulum, Golgi body, lipid and glycogen deposits, secretory granules and surface area of basement membrane were increased. 3) For SEM, the degree of folding and microvilli of surface of mouse epithelial cells was became more and more according to the process of secretory phase, and at D5, implantation time of mouse, the appearance of pinopodes as a specific marker of uterine receptivity was found. The uterine pinopodes of mouse were found in narrow sites at the luminal surface, irregularity and appeared the different stages in the same sample. Therefore, these results indicated that the mouse endometrium was experienced dramatic morphological changes during peri-implantation period.
Journal of the Korean Applied Science and Technology
/
v.37
no.3
/
pp.484-495
/
2020
This study is a mixed surfactant (MimicLipid-MSM1000) that forms the same structure as that of the stratum corneum, sucrose distearate, polyglyceryl-2 dioleate, fermented squalane, ergosterol, 10-hydroxystearic acid, mixture consisting of was synthesized. When using 2~5 wt% of this mixed surfactant, it was possible to make an artificial skin mimetic that forms a multi-layer lamellar structure of 5~30 layers. An emulsion was prepared using this mixed surfactant, and a multi-layered lamellar phase was formed and analyzed mechanically. The appearance of this surfactant was a light brown hard wax, the hydrophilic lipophilic balance (HLB) was 12.53, the critical parameter value was 0.987, and the acid value was 0.13. Stability according to pH change was also stable in acidic (3.8), neutral (7.2) and alkaline (10.8). The particle size of the liquid crystal was found to be the most stable maltese cross lamellar crystalline droplet at 5~25mm. The size of the emulsified particles according to the change in the speed of the homo agitator is 2500 rpm (17.9mm±2.6mm), 3500rpm (12.5mm±2.1mm), 4500rpm (6.2mm±1.8mm) particles were formed. Liquid crystal forming particles were observed through a polarization microscope, and the formation structure of the liquid crystal was precisely analyzed with a scanning electron microscope (cryo-SEM). As an application field, it is expected that it will be widely applicable to the development of various prescriptions, such as various skin care cosmetics, makeup care cosmetics, and scalp protection cosmetics, by using a skin-mimicking surfactant.
Kim, Ki-Woong;Kim, Hyeon-Yong;Park, Sang-Shin;Jeong, Hyo-Seok;Park, Sang-Hoi;Lee, Jun-Yeon;Jeong, Jae-Hwang;Moon, Young-Hahn
BMB Reports
/
v.32
no.3
/
pp.232-238
/
1999
Sex differences in the induction of microsomal cytochrome P-450 (CYP) and the activities of several related enzymes of Sprague-Dawley rats treated with 1-bromopropane (1-BrP) were investigated. Male and female rats were exposed to 50, 300, and 1800 ppm of 1-BrP per kg body weight (6 h a day,S days a week, 8 weeks) by inhalation. The mean body weight of 1-BrP treated groups increased according to the day elapsed, but four and five weeks respectively after the start of the exposure, the mean body weight of male and female rats had significantly reduced in the group treated with 1800 ppm 1-BrP compared with the control group (p<0.01). While the relative weights of liver increased in both sexes, statistical significance in both sexes was found only in the group receiving 1800 ppm/kg of 1-BrP (p<0.01). The total contents of CYP, $b_5$, NADPH-P-450 reductase, NADH $b_5$ reductase, ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-dealkylase (PROD), and p-nitrophenol hydroxylase (pNPH) activities were examined for the possible effects of 1-BrP. No significant changes in the CYP and $b_5$ contents, NADPH-P-450 reuctase, NADH $b_5$ reductase, ethoxyresorufin-O-deethylase (EROD), and pentoxyresorufin- O-dealkylase (PROD) were observed between the control and treated groups. The activity of pNPH increased steadily with the increase in the concentration of 1-BrP in both sexes, but was significantly increased only in the 1800 ppm-treated group of male rats (p<0.05). When Western blottings were carried out with three monoclonal antibodies (MAb 1-7-1, MAb 2-66-3, and MAb 1-98-1) which were specific against CYP1A1/2, CYP2B1/2, and CYP2E1, respectively, a strong signal corresponding to CYP2E1 was observed in microsomes obtained from rats treated with 1-BrP. Glutathione S-transferase (GST) activity and the content of lipid peroxide significantly increased in the treated groups compared with the control group (p<0.05). These results suggest that 1-BrP can primarily induce CYP2E1 as the major form and that GST phase II enzymes play important roles in 1-BrP metabolism, showing sex-dependence in the metabolic mechanism of 1-BrP in the rat liver.
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.8
/
pp.1062-1068
/
2009
Diacylglycerol (DAG) were synthesized by enzymatic esterification of glyceryl monooleate (GMO) and conjugated linoleic acid (CLA) in a shaking water bath. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosa). Effects of reaction time, molar ratio, enzyme road and molecular sieves were studied. Results of normal-phase high performance liquid chromatography (NP-HPLC) analysis were performed. At 1:1, 2:1 and 3:1 (GMO : CLA) molar ratio and Lipozyme TLIM of 20% amount, DAG were produced in 42.6, 54.4 and 54.6 area% in 1 hr, respectively. When different Lipozyme TLIM amounts (2, 5, 10, 20%) were used with 2:1 (GMO : CLA) molar ratio, DAG were produced 21.4 (24 hr), 51.7 (12 hr), 56.2 (6 hr) and 54.4 (1 hr) area%, respectively. The reaction in the absence of molecular sieves increased DAG contents. The maximum DAG concentration conditions were obtained with molar ratio of 2:1 (GMO : CLA), lipase concentration of 10% (of substrate), 10% molecular sieves and reaction time of 6 hours at 55$^{\circ}C$. Under this reaction condition, produced DAG-rich oil was composed of 69 area% DAG, 7.9 area% TAG, 2 area% FFA, and 21.1 area% MAG.
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