• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.70-77
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• 2011

This study was conducted to evaluate the efficacy of human milk fortifier (HMF) on growth and nutritional status in growing rats fed infant formula supplemented with HMF. Three week-old male Sprague-Dawley rats were divided into three groups and fed regular formula (RF), premature formula (PF) and regular formula fortified with HMF (RF+HMF) diets for 3 weeks. There was no significant difference in weight gain among groups. However, a significant increase of food intake was observed in PF and RF+HMF groups compared with RF group. With increasing food intake, the intakes of carbohydrate and protein were significantly increased in PF and RF+HMF groups. The weight of perirenal fat was significantly increased in rats fed RF+HMF; however, the weights of liver, kidney and spleen were not significantly different among groups. Although total lipids, total cholesterol, HDL-cholesterol concentrations of serum were not significantly different among groups, triglyceride was significantly increased in PF group. The triglyceride and total-cholesterol of liver were significantly increased in rats fed regular formula fortified with HMF and PF compared with RF group. Glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), creatinine (Cre) and blood urea nitrogen (BUN) in serum showed no significant difference among groups. The concentration of growth hormone was significantly increased in PF group compared with other groups. The concentration of hemoglobin was significantly increased in rats fed PF and RF+HMF. These results suggest that the supplementation of human milk fortifier in growing rats may promote growth as increasing food intake and lipid contents in tissues and prevent the anemia of infants.

• Journal of Animal Science and Technology
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• v.59 no.7
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• pp.16.1-16.6
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• 2017

Background: This study was performed to investigate the impact of exogenous ghrelin on the pancreatic ${\alpha}$-amylase outputs and responses of pancreatic proteins to ghrelin that may relate to pancreatic exocrine. Methods: Sprague-Dawley male rats (9 weeks old, $300{\pm}10g$) were injected with ghrelin via intraperitoneal (i.p.) infusion at dosage of 0, 0.1, 1.0 and $10.0{\mu}g/kg$ body weight (BW), respectively. The plasma ghrelin and cholecystokinin (CCK) level were determined using enzyme immunoassay kit; the mRNA expression of ghrelin receptor ($GHSR-1{\alpha}$) and growth hormone (GH) receptor were assessed by reverse transcription PCR; the expressions of pancreatic ${\alpha}$-amylase activity, extracellular-signal-regulated kinases (ERK), phosphorylated extracellular-signal-regulated kinases (pERK) and c-Jun N-terminal kinase (JNK) were evaluated by western blotting; moreover the responses of pancreatic proteins to ghrelin were analyzed using the two-dimensional gel electrophoresis system. Results: The exogenous ghrelin (1.0 and $10.0{\mu}g/kg\;BW$) elevated the level of plasma ghrelin (p < 0.05), and suppressed the expression of pancreatic ${\alpha}$-amylase at a dose of $10.0{\mu}g/kg\;BW$ (p < 0.05). No difference in the level of plasma CCK was observed, even though rats were exposed to any dose of exogenous ghrelin. In addition, a combination of western blot and proteomic analysis revealed exogenous ghrelin ($10.0{\mu}g/kg\;BW$) induced increasing the JNK and ERK expressions (p < 0.05) and four proteins such as Destrin, Anionic trypsin-1, Trypsinogen, and especially eukaryotic translation initiation factor 3 in rat pancreas. Conclusions: Taken together, exogenous ghrelin by i.p. infusion plays a role in the pancreatic exocrine secretion via mitogen-activated protein kinase signaling pathway.

• Journal of Preventive Medicine and Public Health
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• v.38 no.3
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• pp.330-336
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• 2005

Objectives : The aim of this study was to investigate the effects of bisphenol A (BPA), an estrogen-like environmental endocrine disrupter, on the placental function and reproduction in rats. The mRNA levels of the placental prolactin-growth hormone(PRL-GH) gene family, placental trophoblast cell frequency and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats ($160g{\pm}20g$) were detected by the presence of the copulatory plug or sperm in the vaginal smear, which marked Day 0 of pregnancy. Pregnant rats were divided into three groups. The control group was intraperitoneally injected with a sesame oil vehicle. The two remaining groups were injected with 50 or 500 mg/kg B.W/day of BPA, resuspended in sesame oil, on either days 7 to 11 or 16 to 20 of pregnancy, with the rats sacrificed on either day 11 or 20, respectively. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction. The hormone concentrations were analyzed by radioimmunoassay, and the frequency of the placental trophoblast cells observed by a histochemical study. Reproductive data, such as the placental weight and litter size, were surveyed on day 20. The fetal weight was surveyed for 4 weeks after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the PRL-GH gene family, such as placental lactogen I, Iv and II, prolactin like protein A, C and Cv, and decidual prolactin-related protein were significantly reduced due to BPA exposure. The mRNA levels of the Pit-1a and b isotype genes, which induce the expression of the PRL-GH gene family in the rat placenta, were also reduced due to BPA exposure. The PL-Iv and PL-II concentrations were reduced in the BPA exposed group. During the middle to last stage of pregnancy (Days 11-20), a high dose of BPA exposure reduced the frequency of spongiotrophoblast cells, which are responsible for the secretion of the PRL-GH hormones. Reproductive data, such as the placental and fetal weights and the litter size, were reduced, but that of the pregnancy period was extended in the BPA exposed compared to the control group. Conclusions : BPA disrupts the placental functions in rats, which leads to reproductive disorders.

• Journal of Life Science
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• v.22 no.4
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• pp.516-523
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• 2012

The purpose of this study was to investigate the effect of exercise and/or L-arginine on abdominal fat, IGF-1 on GH/IGF-1 axis, fibrinogen, and PAI-1 in aged and obese rats. Male Sprague-Dawley rats were treated with a D-galactose aging inducing agent (50 mg/kg) given intraperitoneally for 12 weeks. Thirty-two male Sprague-Dawley rats were treated and divided into four groups: aging-high fat diet group (AG+HF), AG+HF with L-arginine intake group (AG+LA), AG+HF with exercise group (AG+EX), and AG+EX with L-arginine intake group (AG+LA+EX). The experimental rats underwent treadmill training (60 min/day, 6 days/week at 0% gradient) for 12 weeks. L-arginine was given orally (150 mg/kg/day) for 12 weeks. After the experiment, blood was collected from the left ventricle and abdominal fat was extracted. The results showed that GH was significantly increased in AG+EX and AG+AL+EX. IGF-1 was significantly increased in both the AG+AL+EX and AG+EX group ($p$<0.05), while fibrinogen and PAI-1 were not significantly different among the groups. Abdominal fat was significantly decreased in the AG+LA, AG+EX, and AG+LA+EX groups ($p$<0.05) compared with the AG+HF group. In conclusion, this study suggests that exercise alone or L-arginine alone or a combination not only increases the GH and IGF-1 concentration, but also decreases the abdominal fat mass.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

• Applied Microscopy
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• v.41 no.2
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• pp.109-116
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• 2011

Polycystic ovary syndrome (PCOS) is hormonal imbalance condition as the endocrine and metabolic disorder that induces the infertility and various complications in reproductive age women. Estradiol valerate (EV) is used hormone replacement therapy in menopausal women and is reported that excessive administration of EV induces the PCOS. Nerve growth factor (NGF) is the factor to regulate the survival and maturation of developing neuronal cell and is also synthesized in ovary. And NGF is overexpressed in EV-induced polycystic ovary (PCO) as previously reported. Therefore, this study examined the possibility of NGF as can be used the biological marker in diagnosis of PCOS, the hormonal imbalance condition, using PCO and CHO (chinese hamster ovarian) cell lines. The concentration of EV treatment is optimized a 1 mg as not influence on the proliferation of CHO cell but 2 mg and 3 mg of EV treatment have the inhibition effect at initial stage. The morphological change was not observed in CHO cell after dose dependent manner treatment of EV. Expression of NGF mRNA and protein is significantly increased at 30 min after EV treatment in CHO cells compared to that of control. And NGF protein expression is strongly increased in PCO tissue, which observed many follicular cysts compared to normal ovary tissue. Taken together, overexpression of NGF may be act as a molecule to induce an abnormal development of follicle, suggesting that NGF can be used as a biological marker in diagnosis of PCOS.

• Herbal Formula Science
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• v.13 no.2
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• pp.1-16
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• 2005

Objectives: To find out the effects GGTl, an antiobestic drug widely used in clinics, has on the blood-antiobestic index and the toxicity index using the data from the hGHTg obese male rats. We looked closely into both of the two indices because GGTl antiobestic effect can happen not only by pharmacological action, but also by its toxicity. Also, we verified the difference in effect between GGTl and reductil (sibutramine), which has been approved by the FDA of the United States. Methods: After performing the experiments for 8 weeks on the hGHTg obese male rats divided into three groups: the control group, the GGTl group, and the reductil (RD) group, we anesthetized the rats with Diethyl ether and took a 3ml blood sample from the heart. Then, after coagulating the blood in room temperature by using the plasma separator, we centrifuged it for 25 minutes in 3,000rpm using the high-speed refrigerated centrifuge. We kept the separated plasma in a deep freezer at $-80^{\circ}C$, and repeatedly measured the antiobestic index and the toxicity index twice using the hematology biochemistry analyzer. Also, in order to judge the indirect toxicity index, we separated liver from kidney and observed them. Results: When we looked at the results of the analysis of covariance on the measuring elements related to the antiobestic index (TC, HDL, LDL, TG, and GLU), there was no significant difference among the groups in all measuring elements. Also, the results of the analysis of covariance on the two roups (RD group and GGTl group) showed that the p-values had no significant difference under the level of significance 0.05. When we looked at the result of the analysis of covariance on the measuring elements related to the toxicity index (GOT, GPT, GGT, CREA, UA, ALB, and TP), we could see that the p-values in GPT, ALB, and TP have a significant difference among the groups. Also, the results of the analysis of covariance about the measuring elements related to the toxicity index on both groups, RD group and GGTl group, showed no significant difference in the p-values of all of the measuring elements in the two groups, RD and GGTl group. Conclusions: In conclusion, through this experiment, the safety of GGTl has been approved, and although the verification on its medical effect has not been clearly approved, when we consider the fact that it belongs to the same group as reductil, an antiobestic drug approved by the FDA of the United States, we could indirectly verify that GGTl has an antiobestic effect. We believe that when doing a sample design for a future experiment, it needs to be performed on a greater sample size based on the power analysis that needs to be performed primarily in experiments, and a more accurate verification is needed through more systematic experiment plans.

• Journal of Life Science
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• v.16 no.6
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• pp.1014-1028
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• 2006

This study attempted to investigate the developmental alterations of the cerebral cortex. The effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by lectin histochemical methods. To investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, Con A and LCA) were detected with by IHC using ABC kit. In the case of injection of EGEE, the reactions to Con A and LCA were shown in binding phase in the cerebral cortex commonly, and the reactions to PNA, RCA-1 and LCA were shown partially, the number of lectins to be shown reaction were decreased, and there were no reactions to DBA, SBA, BSL-1, RCA-1 and UEA-1. The reaction to Con A was similar to control group during developmental phases. The reaction to LCA was increased in the fetal, suckling, and weanning phases compared with control group. But there were no reactions to SBA and sWGA, the reaction to PNA was decreased in the frontal and occipital cortex and no reaction to sWGA in the fetal phase. There were no reactions to sWGA and PNA in the suckling phase and, no reaction to PNA and sWGA. The reaction to Con A was decreased in the frontal, parietal and occipital cortexes and, the reaction to LCA was decreased in the frontal and occipital cortexes in adult phase. As the results, the effects of EGEE, environmental hormone on the each part of cerebral cortex have shown differences. But, It had deep effect on the differentiation and growth in the cerebral cortex pathologically. In particular, the effect was severe in suckling phase. $Galactosyl-({\beta}-1,3)-N-acetyl-D-galactosamine$,${\beta}-N-acetyl-D-galactosamine$ and ${\beta}-N-acetyl-D-glucosamine$ were decreased while ${\alpha}-D-mannose$ and ${\alpha}-D-glucose$ were increased. It affected the sugar metabloism, and it was severe in fetal and suckling phases.