• KSBB Journal
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• v.14 no.5
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• pp.593-599
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• 1999

In this study, optimal conditions to infect CD34 positive cells containing hematopoietic stem cells obtained from cord blood and bone marrow were found using two different retroviral vectors expressing human growth hormone (hGH) and $\beta$-galactosidase. CD34 positive cells were successfully infected with recombinant retroviruses only when the CD34 positive cells were co-cultured with packaging cells secreting recombinant retroviruses. To find the highest infection efficiency for the gene transfer, CD34 positive cells from cord blood were co-cultured with packaging cells secreting recombinant retroviruses encoding E. coli lacZ gene. The highest infection efficiency was obtained when CD34 positive cells were cultured for 3 days, and then co-culturing was done for another 2 days. When CD34 positive cells from bone marrow were co-cultured with packaging cells secreting recombinant retroviruses encoding hGH gene, the maximum amount of hGH was also secreted at the same conditions found above, i.e. 3 days of culture and 2 days of co-culture. These results show that there are optimal conditions for the gene transfer into hematopoietic stem cells regardless of sources of target cells or retroviral vectors used to infect.

• KSBB Journal
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• v.23 no.3
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• pp.231-238
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• 2008

The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

• Development and Reproduction
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• v.6 no.2
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• pp.97-104
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• 2002

In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.

• Development and Reproduction
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• v.6 no.2
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• pp.117-122
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• 2002

Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX＋Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX＋Oil group and in OVX＋E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX＋Oil and OVX＋E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

• Development and Reproduction
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• v.10 no.1
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• pp.55-61
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• 2006

There is growing concern that dietary soy intake is associated with protection of breast cancer. However, questions persist on the potential adverse effects of the main soy constituent genistein(GS) on female reproductive physiology. In this study, we examined whether prepubertal exposure to GS affected on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. GS(100mg/kg/day) was administrated daily from postnatal day 25(PND 25) to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the day after VO occurred. Gross anatomy and tissue weight were compared to test the GS's effect on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in tissues. Specific radioimmunoassay(RIA) were carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptors(PR), total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a results, advanced VO was shown in the GS group(PND $31.2{\pm}0.6$) compared to the vehicle group (PND $35.3{\pm}0.7$). GS treatment significantly increased wet weight of ovaries and uteri compared to the vehicle group. Increased serum LH levels were also shown in the GS group. Graafian follicles and corpora lutea(CL) were observed only in the ovaries from GS treated animals. Similarly, hypertrophy of luminal and glandular uterine epithelium were found only in the GS group. Collectively, these effects were probably due to the estrogenic effects of GS. In the semi-quantitative RT-PCR studies, the transcriptional activities of PR in both ovary and uterus from GS-treated group were significantly higher than those from the vehicle group. The present studies demonstrated that acute exposure to GS, at levels comparable to the ranges of human exposure, during the critical period of prepubertal stage activates the reproductive system resulting precocious puberty in immature female rats.

• Journal of Dairy Science and Biotechnology
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• v.14 no.1
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• pp.57-69
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• 1996

This study was desiged to carry out an observation study on the cognitive level of health benefitis of fermented milk and subsequntly, on prevention of gastrointestinal diseases for the consumers in Korea. Data used in this study were collected from two different source ; 1) 987 university students living in Seoul, were selected randomly and interviwed induvidual from May 25 to September 30 1994, to investigate the awareness status as a consomer of fermented milk, and 2) health benefits of fermented milk with respect to prevention gastric intestinal diseases such as typhoid fever, paratyphoid fever and bacillary dysentery in Korea with the raw data from the Yearbook of Health and Social Statistics by Ministry of Health and Social Affairs (1976${\sim}$1995) and Ministry of Agriculture, Forestry & Fisheries (1976${\sim}$1995), Republic of Korea. The results of this study were as follows : The effects of femented milk on health appeared to be attributed to taking care of an environment of the gastrointestinal tract regarding diarrhea, constipation and digestion, with showing 75.1% of male and 71.1% of female of university students in Seoul answered positively, The correlation coefficients of statistics between amounts of consumption per capita of fermented milk and incidence rates of gastrointestinal diseases during the period from 1975 to 1994 in the whole country was r = -0.308 (p<0.05). those of Seoul a major comsuming area of fermented milk in the country and Kangwon province, a minor consuming area were r = -0.704 (p<0.01) and r = +0.262 (n.s.), respectively. In conclusion, the results obtained in this study suggest that fermented milk exert its effect on gastrointestinal tract. Furthermore, the results warrant a futher study in the field of preventive medicine and health science with respect to efficacy of fermented milk consumption.

• Journal of Dairy Science and Biotechnology
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• v.16 no.2
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• pp.119-136
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• 1998

The lipids of milk provide energy and many essential nutrients for the newborn animal. They also have distinctive physical properties that affect the processing of dairy products. Milk fat globules mainly consist of neutral lipids like triacylglycerols, whereas the globule membranes contain the complex lipids mostly, Phospholipids are a small but important fraction of the milk lipids and are found mainly in the milk fat globule membrane and other membranous material in the skim-milk phase. The milk fats of ruminant animals are characterized by the presence of relatively high concentrations of short-chain fatty acids, especially butyric and hexanoic acids, which are rarely found in milks of non-ruminants. The fatty acids of milk lipids arise from de novo synthesis in the mammary gland and uptake from the circulating blood. The fatty acid compositions of milks are usually complex and distinctive, depending on the nature of the fatty acids synthesized de novo in the mammary gland and those received from the diet in each species. The content and composition of milks from different species vary widely; presumably, these are evolutionary adaptations to differing environments. The actual process by which these globules are formed is unkonwn, but there are indications that triglyceride-containing vesicles which bleb from endoplasmic reticulum may serve as nucleation sites for globules. Recent studies on milk have centred on the manipulation of milk lipids to increase specific fatty acids, i.e. 20-carbon omega-3 fatty acids (eicosapentaenoic acid 20:5n3, decosahexaenoic acid 22:6n3) from marine sources because the fatty acids are closely associated with a decreased risk of coronary heart disease.

• Journal of fish pathology
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• v.10 no.2
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• pp.97-111
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• 1997

In the winter of 1995, mass mortality occurred in cultured flounder, Paralichthys olivaceus in Gurongpo, Kyoungbuk, Korea. From the observations of moribund and dead fish, parasitic ciliates, which were shown as white spots to the naked eye, were considered to be involved in the mass mortality. From heavily infected flounders, histopathological, morphological and biological characterization of these ciliates were carried out. In the histological observation, many ciliates were found under the epithelia of gill filaments and skin, and caused hyperplasia of epithelial and mucus cells at the infected areas. The ciliates found on the body surface, fins and gills were very similar to Cryptocaryon irritans. However the ciliates showed two different patterns of reproductian, i.e., typical form(palintomy)and atypical form(budding plus multiple fission) at $16^{\circ}C$ of water temperature. The occurrence ratio between typical and atypical form was about 3:2. Tomitogenesis takes 8-14 days in the typical and 13-15 days in the atypical form. In the viability test at different temperatures and salinities, the typical form died below 30‰ at $12^{\circ}C$, below 20‰ at $16^{\circ}C$, below 15‰ at $20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. On the other hand, the atypical form died below 20‰ at $12^{\circ}C$, below 15‰ at 16-$20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. The results suggested that the atypical has better viability at low salinity than that of the typical at low temperatures. In the excystment time and success rates of excystment according to temperatures, the typical form showed 8 days, 30% at $12^{\circ}C$ : 6.5 days, 50%, at $16^{\circ}C$ : 5.5 days, 75% at $20^{\circ}C$ : and 7 days, 10% at $24^{\circ}C$, respectively. On the other hand, the atypical form showed 15.5 days at $12^{\circ}C$ : 14 days, 76.6% at $16^{\circ}C$ : 12 days, 72.2% at $20^{\circ}C$ : 10 days 31.6% at $24^{\circ}C$, respectively. The results suggested that the atypical form had longer excystment time than that of the typical form at any temperature and showed better stability at low temperatures.

• Korean Journal of Food Science and Technology
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• v.5 no.1
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• pp.6-15
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• 1973

In an attempt to develop composite flours based on raw materials available in Korea, six domestic resources were investigated with respect to their physical and chemical properties, nutritive value and economy. The results are summarized as follows: 1) Flours from naked barley, sweet potato, potato, corn, defatted rice bran and defatted soybean were compared in respect of their proximate composition, thiamine content and protein score. 2) In color comparison, naked barley and defatted soy flours were comparable to wheat flour whereas corn and sweet potato flours were a little inferior. In raising power, naked barley and defatted soy flours were similar to wheat flour and others were inferior. 3) In maximum viscosity of flours by amylograph, naked barley, corn and potato flours were higher than wheat flour and others were lower. In viscosity in cooling, corn flour was similar to wheat flour and naked barley and potato flours were lower. Addition of $10{\sim}20%$ defatted soy flour into other flours generally caused the lowering of viscosity. Addition of emulsifiers such as glyceryl monostearate, calcium stearyl lactylate, Methocel and Emulthin into the flours manifested different effects on the amylogram. 4) In nutritional test by rat, diets consisting of naked barley, sweet potato, potato or corn flours, each containing defatted soy flour (at 10% level with exception of 20% for sweet potato flour) and naked barley flour fortified with lysine were similar to wheat flour in digestibility, but were superior in NPU and biological value. 5) In price estimation, sweet potato and corn flours were competitive with wheat flour, but naked barley flour was a little expensive. On the other hand, barley and sweet potato are promising in terms of domestic production.

• BMB Reports
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• v.36 no.1
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• pp.82-94
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• 2003

Although debates still exist whether Helicobacter pylori infection is really class I carcinogen or not, H. pylori has been known to provoke precancerous lesions like gastric adenoma and chronic atrophic gastritis with intestinal metaplasia as well as gastric cancer. Chronic persistent, uncontrolled gastric inflammations are possible basis for ensuing gastric carcinogenesis and H. pylori infection increased COX-2 expressions, which might be the one of the mechanisms leading to gastric cancer. To know the implication of long-term treatment of antiinflammatory drugs, rebamipide or nimesulide, on H. pylori-associated gastric carcinogenesis, we infected C57BL/6 mice with H. pylori, especially after MNU administration to promote carcinogenesis and the effects of the long-term administration of rebamipide or nimesulide were evaluated. C57BL/6 mice were sacrificed 50 weeks after H. pylori infection. Colonization rates of H. pylori, degree of gastric inflammation and other pathological changes including atrophic gastritis and metaplasia, serum levels and mRNA transcripts of various mouse cytokines and chemokines, and NF-${\kappa}B$ binding activities, and finally the presence of gastric adenocarcinoma were compared between H. pylori infected group (HP), and H. pylori infected group administered with long-term rebamipide containing pellet diets (HPR) or nimesulide mixed pellets (HPN). Gastric mucosal expressions of ICAM-1, HCAM, MMP, and transcriptional regulations of NF-${\kappa}B$ binding were all significantly decreased in HPR group than in HP group. Multi-probe RNase protection assay showed the significantly decreased mRNA levels of apoptosis related genes and various cytokines genes like IFN-$\gamma$, RANTES, TNF-$\alpha$, TNFR p75, IL-$1{\beta}$ in HPR group. In the experiment designed to provoke gastric cancer through MNU treatment with H. pylori infection, the incidence of gastric carcinoma was not changed between HP and HPR group, but significantly decreased in HPN group, suggesting the chemoprevention of H. pylori-associated gastric carcinogenesis by COX-2 inhibition. Long-term administration of antiinflammatory drugs should be considered in the treatment of H. pylori since they showed the molecular and biologic advantages with possible chemopreventive effect against H. pylori-associated gastric carcinogenesis. If the final concrete proof showing the causal relationship between H. pylori infection and gastric carcinogenesis could be obtained, that will shed new light on chemoprevention of gastric cancer, that is, that gastric/cancer could be prevented through either the eradication of H. pylori or lessening the inflammation provoked by H. pylori infection in high risk group.