This study was designed to investigate the effects of ethyl acetate(EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle on cholesterol and lipofuscin(LF) accumulations, acetylcholine(ACh) and its related enzyme activities such as choline acetyltransferase(ChAT) and acetylcholinesterase(AChE), and monoamine oxidase-B(MAO-B) activity, which destroyed the catecholamine related neurotransmitters in brain membranes of Sprague- Dawley (SD) rats. Male SD rats were fed basic diets (control group) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45 days. Cholesterol accumulations in mitocholndria and microsomes were significantly inhibited (11.8-12.1% and 9.6-13.0%, respectvely) in EtOAc-50 and EtOAc-100 groups. ACh levels and ChAT activities were significantly increased about 10% in membranes of EtOAc-100 group compared with control group. AChE activities were significantly increased about 8 -12% in membranes of EtOAc-50 and EtOAc-100 groups compared with control group. MAO-B activities were significantly inhibited about 10% in membrane of EtOAc-l00 group compared with control group. These results suggest that ethyl acetate fraction of pine needle may play an effective role in inhibiting cholesterol and improving a membrane fluidity, and learning and memory impairments. (Korean J Nutrition 37(2): 95 -99, 2004)
This study was conducted to evaluate the effect of dietary antioxidant and energy density on performance and antioxidative status in transition cows. Forty cows were randomly allocated to 4 dietary treatments in a $2{\times}2$ factorial design. High or low energy density diets (1.43 or 1.28 Mcal $NE_L$/kg DM, respectively) were formulated with or without antioxidant (AOX, a dry granular blend of ethoxyquin and tertiary-butylhydroquinone; 0 or 5 g/cow per d). These diets were fed to cows for 21 days pre-partum. During the post-partum period, all cows were fed the same lactation diets, and AOX treatment followed as for the pre-partum period. Feeding a high energy diet depressed the DMI, milk yield, and 4% fat-corrected milk (FCM) of cows. However, AOX inclusion in the diet improved the milk and 4% FCM yields. There was an interaction of energy density by AOX on milk protein, milk fat and total solids contents. Feeding a high energy diet pre-partum increased plasma glucose and ${\beta}$-hydroxybutyrate, whereas dietary AOX decreased plasma ${\beta}$-hydroxybutyrate value during the transition period. There were also interactions between time and treatment for plasma glutathione peroxidase activity and malondialdehyde content during the study. Cows fed high energy diets pre-partum had higher plasma glutathione peroxidase activity 3 days prior to parturition, compared with those on low energy diets. Inclusion of AOX in diets decreased plasma glutathione peroxidase activity in cows 3 and 10 days pre-partum. Addition of AOX significantly decreased malondialdehyde values at calving. Energy density induced marginal changes in fatty acid composition in the erythrocyte membrane 3 days post-partum, while AOX only significantly increased cis-9, trans-11 conjugated linoleic acid composition. The increase in fluidity of the erythrocyte membrane was only observed in the high energy treatment. It is suggested that a diet containing high energy density pre-partum may negatively affect the anti-oxidative status, DMI and subsequent performance. Addition of AOX may improve the anti-oxidative status and reduce plasma ${\beta}$-hydroxybutyrate, eventually resulting in improved lactation performance; the response to AOX addition was more pronounced on the high energy diet.
Park, Cheol-Ho;Kim, Sung-Won;Hwang, You-Jin;Kim, Dae-Young
Journal of Embryo Transfer
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v.27
no.2
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pp.107-111
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2012
Miniature pig sperm cryopreservation is continually researched in biotechnology for breed conservation and reproduction. It is important to control the temperature at each stage of cryopreservation and cryoprotectant. It is also necessary to find the optimal cryoprotectant concentration and chemical elements of the extender. Recently, many studies have used various cryoprotectant materials, such as dimethyl sulphoxide (DMSO), ethylene glycol (EG), antifreeze protein (AFP), amides, and glycerol. Glycerol is a commonly used cryoprotectant. However, glycerol has critical cytotoxic properties, including osmotic pressure and it can cause irreversible damage to live cells. Therefore, We focused on membrane fluidity modifications can reduce cell damage from freezing and thawing procedures and evaluated on the positive effects of trehalose to the viability, chromatin integrity, and motility of boar sperm. Miniature pig sperm was separated from semen by washing with modified- Modena B (mMB) extender. After centrifugation, the pellet was diluted with the prepared first extender. This experiment was designed to compare the effects that sperm cryopreservation using two different extenders has on sperm chromatin. The control group used the glycerol only and it was compared with the glycerol and glycerol plus trehalose extender. Sperm viability and motility were evaluated using WST1 assays and computer-assisted semen assays (CASA). Chromatin structure was examined using acridine orange staining. For the motility descriptors, trehalose caused a significant (p<0.01) increase in total motility ($57.80{\pm}4.60%$ in glycerol vs. $75.50{\pm}6.14%$ in glycerol + trehalose) and progressive ($51.20{\pm}5.45%$ in glycerol vs. $70.74{\pm}8.06%$ in glycerol + trehalose). A significant (p<0.05) increase in VAP ($42.70{\pm}5.73{\mu}m/s$ vs. $59.65{\pm}9.47{\mu}m/s$), VSL ($23.06{\pm}3.27{\mu}m/s$ vs. $34.60{\pm}6.58{\mu}m/s$), VCL ($75.36{\pm}11.36{\mu}m/s$ vs. $99.55{\pm}12.91{\mu}m/s$), STR ($54.4{\pm}2.19%$ vs. $58.0{\pm}1.63%$), and LIN ($32.2{\pm}2.05%$ vs. $36.0{\pm}2.45%$) were also detected, respectively. The sperm DNA fragmentation index was 48.8% to glycerol only and 30.6% to glycerol plus trehalose. Trehalose added group showed higher percentages of sperm motility, stability of chromatin structure than glycerol only. In this study, we suggest that trehalose is effective in reducing freezing damage to miniature pig sperm and can reduce chromatin damage during cryopreservation.
Recently, there were many studies not only to enhance drug delivery effect but to reduce side effect. Drug delivery system(DDS) is able to improve efficiency with decreasing side effect of drug dosage. Among these application fields, DDS is often used as the method of drug dosage into the epidermic skin. We investigated characters of transdermal therapeutic system(TTS) and the skin permeability of that with applying DDS. We investigated the permeation of xanthan gum containing drug in rat skin using borizontal membrane cell model. Permeation properties of materials were investigated for water-soluble drug with oxiniacic acid and also for lipophilic drug with clofibrate. The permeation rate of lipophilic drug was found to be faster than that of water-soluble drug in vitro. The rate differences of both water-soluble drug and lipophilic drug according to drug content were negligible. We used glycerin, PEG 600 and oleic acid as enhancers. These results showed that skin permeation rate of each drug across the composite was mainly dependent on the property of base and chemical property of drug etc.. Proper selection of the polymeric materials which resemble and enhance properties of the delivering drug was found to be important in controlling the skin permeation rate. This result suggests a possible use of natural polymer base as a transdermal delivery system of antihyperlipoproteinemic agent.
Rice (Oryza sativa) cultivars show an impairment of growth and development in response to abiotic stresses such as drought, salinity, heat and cold at the early seedling stage. The tolerance to heat stress in plants has been genetically modulated by the overexpression of heat shock transcription factor genes or proteins. In addition to a high temperature-tolerance that has also been altered by elevating levels of osmolytes, increasing levels of cell detoxification enzymes and through altering membrane fluidity. To examine the heat tolerance in transgenic rice plants, three OsOPT10 overexpressing lines were characterized through a physiological analysis, which examined factors such as the electrolyte leakage (EL), soluble sugar and proline contents. We further functionally characterized the OsOPT10 gene and found that heat induced the expression of OsOPT10 and P5CS gene related proline biosynthesis. It has been suggested that the expression of OsOPT10 led to elevated heat tolerance in transgenic lines.
Liposome as a carrier of topotecan (TPT), a promising anticancer drug, has been reported in attempt to improve the stability and antitumor activity of TPT. However, the biodistr ibution pattern of TPT liposome in vivo and PEG-modified liposome containing TPT have not been studied systemically. In this paper, the in vitro stability and in vivo biodistribution behavior of several liposomes containing TPT with different lipid compositions and PEG-modification were studied. Compared with the 'fluid' liposome (S-Lip) composed of soybean phosphatidylcholine (SPC), the 'solid' liposome (H-Lip) composed of hydrogenated soybean phosphatidylcholine HSPC decreased the leaking efficiency of TPT from liposome and enhanced the stability of liposome in fetal bovine serum (FBS) or human blood plasma (HBP). The results of biodistribution studies in S$_{180}$ tumor-bearing mice showed that liposomal encapsulation increased the concentrations of total TPT and the ratio of lactone form in plasma. Compared with free TPT, S-Lip and H-Lip resulted in 5- and 19- fold increase in the area under the curve (AUC$_{0\rightarrow\propto}$), respectively. PEG- modified H-Lip (H-PEG) showed 3.7-fold increase in AUC$_{0\rightarrow\propto}$ compared with H-Lip, but there was no significant increase in t$_{1/2}$ and AUC$_{0\rightarrow\propto}$ for PEG-modified S-Lip (S-PEG) compared with S-Lip. Moreover, the liposomal encapsulation changed the biodistribution behavior, and H-Lip and H-PEG dramatically increased the accumulation of TPT in tumor, and the relative tumor uptake ratios were 3.4 and 4.3 compared with free drug, respectively. There was also a marked increase in the distribution of TPT in lung when the drug was encapsulated into H-Lip and H-PEG. Moreover, H-PEG decreased the accumulation of TPT in bore marrow compared with unmodified H-Lip. All these results indicated that the membrane fluidity of liposome has an important effect on in vitro stability and in vivo biodistribution pattern of liposomes containing TPT, and PEG-modified 'solid' liposome may be an efficient carrier of TPT.
The protective effects of sea tangle (Laminaria japonica) extract and fucoidan components on the attack of oxygen radicals in kidney were studied, Sprague-Dawley (SD) male rats (210 $\pm$ 5 g) were with fed experimental diets of Dasi-Ex group (sea tangle extract powder of $4.0\%$ added to control diet), Euco-I, II and III groups (fucoidan powder of 1, 2 and $3\%$, respectively, added to Dasi-Ex group) for 45 days, Hydroxyl radical formations were significantly decreased ($10\~15\%$ and $15\~30\%$) in mitochondria and microsomes of Dasi-Ex and Fuco-I, II, III groups compared with control group. Hydrogen peroxide formations were also significantly decreased ($10\~15\%$) in microsomes of Dasi-Ex and Fuco-I, II, III groups compared with control group. Significant differences in mitochondrial basal oxygen radical (BOR) and microsomal induced oxygen radical (IOR) formations of Dasi-Ex and Fuco-I groups could not be obtained, but mitochondrial BOR and microsomal IOR formations were significantly decreased ($12\~15\%$ and $13\~14\%$) in Fuco-II and III groups compared with control group. BOR formations were significantly decreased ($12\~25\%$) in microsomes of Dasi-Ex and Fuco-I, II, III groups, and IOR formations were also significantly decreased ($10\~15\%$) in mitochondria of Fuco-I, II, III groups compared with control group, Significant differences in mitochondrial Mn-SOD activities of Dasi-Ex group could not be obtained, but mitochondrial Mn-SOD activities were dose-dependently increased by $8\%,\;16\%$ and $36\%$ in Fuco-I, II and III groups compared with control group, Mn-SOD activities an microsome were significantly increased about $20\%$ in Dasi-Ex group, while they were remarkably increased about $40\%$ in Fuco-I, II and III groups compared with control group. lipid peroxide contents were significantly decreased about $15\%$ and $15\~25\%$ in mitochondria and microsomes of Fuco-II and III groups. Membrane fluidities resulted in marked increases ($20\~35\%$ and $17\~24\%$) in mitochondria and microsomes of Dasi-Ex and Fuco-I, II and III groups. These results suggest that administrations of fucoidan added to sea tangle may play a pivotal role in attenuating attack of oxygen radicals in kidney.
Cholesterol is the precursor of various steroid hormones, bile acid, and vitamin D with functions related to regulation of membrane permeability and fluidity. However, the presence of excess blood cholesterol may lead to arteriosclerosis and hypertension. Moreover, dietary cholesterol may affect blood cholesterol levels. Generally, cholesterol determination is performed by spectrophotometric or chromatographic methods, but these methods are very time consuming and costly, and require complicated pretreatment. Thus, the development of a rapid and simple analysis method for measuring cholesterol concentration in food is needed. Multi-walled carbon nanotube (MWCNT) was functionalized to MWCNT-$NH_2$ via MWCNT-COOH to have high sensitivity to $H_2O_2$. The fabricated MWCNT-$NH_2$ was attached to a glassy carbon electrode (GCE), after which Prussian blue (PB) was coated onto MWCNT-$NH_2$/GCE. MWCNT-$NH_2$/PB/GCE was used as a working electrode. An Ag/AgCl electrode and Pt wire were used as a reference electrode and counter electrode, respectively. The sensitivity of the modified working electrode was determined based on the amount of current according to the concentration of $H_2O_2$. The response increased with an increase of $H_2O_2$ concentration in the range of 0.5~500 ${\mu}M$ ($r^2$=0.96) with a detection limit of 0.1 ${\mu}M$. Cholesterol oxidase was immobilized to aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, and toyopearl beads. The immobilized enzyme reactors with aminopropyl glass beads and CNBr-activated sepharose showed linearity in the range of 1~100 ${\mu}M$ cholesterol. Na-alginate and toyopearl beads showed linearity in the range of 5~50 and 1~50 ${\mu}M$ cholesterol, respectively. The detection limit of all immobilized enzyme reactors was 1 ${\mu}M$. These enzyme reactors showed high sensitivity; especially, the enzyme reactors with CNBr-activated sepharose and Na-alginate indicated high coupling efficiency and sensitivity. Therefore, both of the enzyme reactors are more suitable for a cholesterol biosensor system.
Journal of the Korean Society of Food Science and Nutrition
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v.32
no.2
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pp.269-277
/
2003
Effects of soy protein concentrate (SPC) containing isoflavone and casein diets on plasma phospholipid (PLs)-fatty acid patterns were investigated in 7-& 40- wk old female rats. Diets containing 16% SPC (soy/young:SY, soy/old: SO) and casein (casein/young : CY, casein/ old: CO) supplemented with 0.5% cholesterol were fed for 4 wks. Fatty acid compositions of plasma PLs were determined by TLC and GLC. Compared to the dietary protein effects, age effects on serum lipids were more profound. The levels of total cholesterol (Chol.), triglyceride, HDL-Chol., (LDL+VLDL)-Chol. and atherogenic index (AI) were higher in older groups (OC & OS) than younger groups (YC &. YS). Soy groups had higher Ell)L-Chol. level and lower (LDL+ VLDL)-Chol. and AI, compared with casein groups. The compositions of C22:0, Cl8:1 $\omega$9 and sum of MUFA in plasma PLs were significantly higher in casein group (CY & CO) than soy group (SY & SO), but those of sum of SFA were higher in soy group. The compositions of C22:0, Cl8:1 $\omega$9, C22:1, Cl8:3$\omega$3 and C22:4$\omega$6 were higher and those of C22:6$\omega$3, sum of $\omega$3, Cl8: 2$\omega$6 C20:4$\omega$6, sum of $\omega$6 and sum of PUFA were lower in plasma PLs of younger rats. The average P/S and $\omega$3/$\omega$6 ratio in older group was higher. The $\Delta$-7 desaturation index (16:0⇒16:1$\omega$7) and $\Delta$-9 desaturation index (18:0⇒18:1$\omega$9) were lower in soy group than casein group, while $\Delta$-6 and $\Delta$-5 desaturation index were not affected by dietary protein. The $\Delta$-4 desaturation index (22:4$\omega$6⇒22:5$\omega$6) were higher and elongation index (20:4$\omega$6⇒22:4$\omega$6) were lower in older group. The ratio of the products of $\omega$3 fatty acid series (Cl8:3) was significantly higher in older group, which indicated that age affected the plasma PUFA metabolism. On the other hand, older rats had higher serum cholesterol level compared with younger rats. Taken together, these changes in fatty acid composition might cause minimal changes in tile membrane fluidity induced by the increase serum cholesterol level.
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