• Journal of Embryo Transfer
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• v.19 no.3
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• pp.219-227
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• 2004

This study was conducted to develop a serum-free, defined medium of IVM of pig oocytes. The TCM-199 with supplemented with polyvinylalcohol(PVA), polyvinylpyrrollidone(PVP) and porcine follicular fluid(pFF) were used as basal medium. The effects of the these additives on the rates of maturity and development under in-vitro fertilization and in vitro culture were examined and subsequently considered on the possibilities be sustituted for the bovine serum albumin(BSA). Maturation rate of pig oocytes in IVM media containing PVA(82.4％), pFF(89.4％) and BSA(90.0％) were significantly higher(P<0.05) than that of PVP(78.6％). Cleavage rate after IVF of PVP(64％) was significantly lower(P<0.05) than these of PVA(73％), pFF(77％) and BSA(73％) supplements. in vitro development rates to morulae and blastocyst on PVP(54％) were also significantly lower(P<0.05) than these of the supplements of PVA(63％), pFF(69％) and BSA(65％). In comparison of maturation and fertilization rates of pig oocytes in each supplements, the maturity rates of PVA(82.4％), pFF(89.4％) and BSA(90.0％) were significantly lower(P<0.05) than that of PVP(72.4％) and while, the fertilization rates of pFF(87.1％) and BSA(89.1％) were significantly higher(P<0.05) than these of PVA(78.0％) and PVP(70.6％). It may be concluded that PVA and pFF can be substituted far BSA in medium for culturing pig oocytes; however, it may be considered that PVP were limited to for BSA in the in vitro culture of the embryos.

• Journal of Aquaculture
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• v.7 no.1
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• pp.63-76
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• 1994

The effects of different dietary lipids on growth, body chemical composition, and nutrient partition of the Korean rockfish (43 g of initial mean weight) were studied. Fish were fed three experimental diets containing 1) $4.5\%$ squid liver oil, 2) $1\%$ squid liver oil and $3.5\%$ beef tallow, and 3) $1\%$ squid liver oil and $3.5\%$ soybean oil. Growth and feed efficiency were detarmined during a 19-week feeding period at water temperature of $15.5^{\circ}C$, and at the end of which body chemical compositions were analyzed. Remained fish were fasted for 45 days, and four fish were taken at 6, 12 and 24 hours, and 3, 15 and 45 days after the fasting to determine tissue compositions and hematological changes. Growth, feed efficiency, protein and lipid retention efficiency, hepatosomatic index(HSI), viscerosomatic index (VSI), and chemical composition of whole body and dorsal muscle were not affected by the different dietary lipids. Liver moisture content of fish fed squid liver oil diet was higher than that of fish fed beef tallow or soybean oil diet. Liver lipid content of fish fed beef tallow diet was higher than that of fish fed the other diets. Fatty acid composition of dorsal muscle and liver were affected by the different dietary lipids ; high levels of 20 : 5n-3 and 22 : 6n-3 from fish fed the diet containing squid liver oil, 18 : 1 from fish fed the diet containing beef tallow, and high 18 : 2n-6 and 18 : 3n-3 from fish fed the diet containing soybean oil were observed. Both HSI and VSI of fish fed three diets decreased with time after the begining of starvation. Liver glycogen did not change during the first 15 days of starvation and decreased thereafter, and that was not affected by the different dietary lipids. Lipid and protein contents in the dorsal muscle of fish decreased up to 15 days of starvation and remained unchanged thereafter, these were not different from each other. Glucose, free fatty acid, triglyceride and phospholipid concentrations in Fish serum were varied for the first 15 days of starvation, after that the concentrations of fish serum remained relatively stable in all the treatment groups with prolonged starvation. The results indicate that Korean rockfish can utilize fish oil, animal fat or vegetable oil equally as energy source when n-3HUF A is adquate.

• Korean Journal of Food Science and Technology
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• v.42 no.5
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• pp.586-592
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• 2010

This study investigated the effects of Stevia rebaudian bertoni leaf extract on lipid profiles in C57BL/6J mice, as compared to stevioside. Fourty mice were divided into four groups: NC (normal diet and DW), HC (high fat diet and DW), HLSV (high fat diet and stevia leaf extract, 1 mL/kg/day), and HSS (high fat diet and stevioside, 1 mL/kg/day). Serum triglyceride concentrations were significantly lower in the HLSV and HSS groups than in the HC group. Epididymal fat pad weights were significantly lower in the HLSV and HSS groups than in the HC group. We also evaluated the serum and liver carnitine levels (NEC, AIAC, ASAC, TCNE) of all the groups. These results were supported by the mRNA expression of enzymes related to lipid metabolism (ACC, PPAR${\alpha}$, ACS, CPT-I) assessed by RT-PCR. Overall, the results show the antiobesity effects of stevia leaf extract as compared to stevioside in high fat diet induced obese mice.

• Development and Reproduction
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• v.5 no.1
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• pp.73-79
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• 2001

Recently many studies have reported that total and bioavailable androgens reduced in male and female athletes and that physical exercise reduces the body weight and increases the reproductive abnormalities such as oligomenorrhea, anovulation, inadequate luteal phase, and delayed puberty in women by the inhibition of the hypothalamic-pituitary-gonadal (HPG) axis . In addition, high mileage endurance 겨nning, psychological stress, and military endurance training in men also reduce the secretion of reproductive hormones. To investigate the efffcts of physical endurance exercise on the secretion of reproductive hormones in men, androgenic hormones from adrenal glands and testis were measured in serum by the conventional radioimmunoassays after long-term (more than3 months), short-term (1 week), and acute (1${sim}$2 hours) physical exercises. Androgenic hormones from adrenal glands and testis such as total testosterone (TT), free testosterone (fT), dehydroepiandrosterone (DHEA), and androstenedione (A) decreased after thesestrenuous endurance trainings, whereas ACTH, cortisol, and dehydroepiandrosterone sulfetes (DHEAS) increased. Conadotropins (LH and FSH) were not idluenced by the physical exercises. Based upon the present results, we assume that the decrease in adrenal and testicular androgens by physical endurance exercises might be associated with the reproductive abnormalities in athletes by unknown factor(s) in addition to the HPG axis disturbance.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

• The Korean Journal of Nuclear Medicine
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• v.36 no.6
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• pp.333-343
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• 2002

Purpose: A new linker, hydrazino nicotinamide (HYNIC), was recently introduced for labelling of liposome with $^{99m}Tc$. In this study we synthesized HYNIC derivatized PEG (polyethylene glycol)-liposomes radiolabeled with $^{99m}Tc$. Materials and Methods: In order to synthesize HYNIC-DSPE (distearoyl phosphatidyl ethanolamine) which is a crucial component for $^{99m}Tc$ chelation, first of all succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized from 6-chloronicotinic acid by three sequential reactions. A DSPE derivative of succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was transformed into HYNIC-DSPE by HCI/dioxane. HYNIC-PEG-liposomes were prepared by hydration of the dried lipid mixture of EPC (egg phosphatidyl choline): PEG-DSPE : HYNIC-DSPE:cholesterol (1.85:0.15:0.07:1, molar ratio). The HYNIC-PEG-liposomes were labeled with $^{99m}Tc$ in the presence of $SnCl_2{\cdot}2H_2O$ (a reducing agent) and tricine (a coligand). To investigate the level of in vivo transchelation of $^{99m}Tc$ in the liposomes, the $^{99m}Tc$-HYNiC-PES-liposomes were incubated with a molar excess of DTPA, cysteine or glutathione solutions at $37^{\circ}C$ for 1 hour. The radiolabeled liposomes were also incubated in the presence of human serum at $37^{\circ}C$ for 24 hours. Results: 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized with 77.3% overall yield. The HYNIC concentration in the PEG-coated liposome dispersion was 1.08 mM. In condition of considering the measured liposomal size of 106 nm, the phospholipid concentration of $77.5\;{\mu}mol/m{\ell}$ and the liposomal particle number of $5.2{\times}10^{14}$ liposomes/ml, it is corresponded to approximate 1,250 nicotinyl hydrazine group per liposome in HYNIC-PEG-liposome. The removal of free $^{99m}Tc$ was not necessary because the labeling efficiency were above 99%. The radiolabeled liposomes maintained 98%, 96% and 99%, respectively, of radioactivity after incubation with transchelators. The radiolabeled liposomes possessed above 90% of the radioactivity in serum. Conclusion: These results suggest that the HYNIC can be synthesized easily and applied in labelling of PEG-liposomes with $^{99m}Tc$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.935-942
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• 2000

This study was designed to investigate the effect of flower of Pueraria lobata on liped peroxidation and activities of alcohol metabolic enzymes in alcohol-treated rats. Male Spra gue-Dawley rats were given 25% ethanol (Alcohol), 25% ethanol and 5 mg tectorigenin/kg B.W.(Alc.-Tec), 25% ethanol and 5mg kaikasaponin III/kg B.W. (Alc-Kai). The contents of serum total lipid, triglyceride and phospholipid were increased by ethanol treatment and were lower in the Alc.-Tec and Alc.-Kai group than in the Alcohol group. Decreased serum HDL-cholesterol by alcohol treatment was recovered by tectorigenin and kaikasaponin III. Microsomal cytochrome P-450, aniline hydroxylase and aminopyrine N-demethylase activities were increased by ethanol and were lower in the Alc. Tec and Alc.-Kai group than in the Alcohol group. Activity of hepatic alcohol dehydrogenase was increased by ethanol and was higher in the Alc.-Tec and Alc.-Kai group than in the Alcohol group. Microsomal ethanol oxidizing system activity was higher in Alc.-Tec group than in the other group. No significant difference was found in catalase activity among treatment groups. These data indicate that tectorigenin and kaikasaponin III were effected alcohol metabolic enzyme system and the liver damage associated with chronic ethanol consumption.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.151-157
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• 2003

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

• Clinical and Experimental Reproductive Medicine
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• v.33 no.1
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• pp.15-23
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• 2006

Objective: Pituitary adenylate cyclase-activating polypeptide (PACAP), a novel hypothalamic neuropeptide, has been suggested to play a role in ovarian folliculogenesis. The present study evaluated the effect of PACAP on the growth of preantral follicles. Methods: Preantral follicles were mechanically isolated from ovaries of 21-day-old rats and cultured in groups for 3 days in serum-free medium in the absence or presence of PACAP-38 ($10^{-6}M$). Results: Treatment with PACAP-38 resulted in an increase in follicle diameter by 75% whereas treatment with follicle stimulating hormone (FSH) increased follicle diameter by 65%. PACAP-38 treatment enhanced the granulosa cell proliferation as measured by thymidine incorporation analysis. Furthermore, the production of progesterone by cultured granulosa cells and GFSHR-17 cell line was stimulated by PACAP-38. Interestingly, PACAP enhanced FSH action on stimulation of SF-1 and aromatase gene expression. Conclusion: The present results demonstrate that PACAP stimulated preantral follicle growth by potentiating proliferation and by stimulating steroidogenesis.

• Journal of Life Science
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• v.19 no.5
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• pp.664-670
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• 2009

The ${\beta}$3-adrenergic receptor (ADRB3) is expressed mainly in visceral adipose tissue and is thought to contribute to lipolysis and the delivery of free fatty acids to the portal vein. This study was aimed at evaluating the association between high-density lipoprotein cholesterol (HDL-C) and ADRB3 genetic polymophisms. A total of 991 healthy examinees who were examined in a university hospital, located in Busan City, between May and December 2006 were enrolled in this study. Height, weight, body mass index, waist circumference, and systolic and diastolic blood pressures of the subjects were examined. Intravenous concentrations of fasting blood glucose, total cholesterol, HDL-C, low-density lipoprotein cholesterol, and triglyceride were also measured. After extracting DNA from the subjects, mutations of the +188T>C (Trp64Arg) of exon 1 and +3893T>C of intron 2 on the ADRB3 gene were genotyped using the single base extension method. We have identified a novel mutation of ADRB3 that is located in intron 2. The frequency of its minor allele was 0.164. Both the +188T>C mutation of exon 1 and +3893T>C mutation of intron 2 were significantly associated with HDL-C. The mean concentration of serum HDL-C was significantly lower in the presence of their minor allele 'C'. These results suggest that both mutations of +188T>C of exon 1 and +3895T>C of intron 2 have significant associations with HDL-C in the Korean population.