• Korean Journal of Animal Reproduction
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• v.18 no.1
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• pp.1-6
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• 1994

The effects of exogenous estrogen on growth, mammary development, and the age at which the first estrous symptoms are detected were investigated for the first time in Korean native heifers at 6 months of age. At 90 days after treatment, weight gains of heifers implanted with 10 mg of estradiol were not different from those of the control heifers. However, weight gams of heifers implanted with 20 mg of estradiol were significantly increased at 90 days by 18%, relative to control values. At 180 days, cumulative weight gains of heifers implanted with either 10 mg or 20 mg of estradiol averaged 6.7% or 17.8%, respectively, greater than controls. But these values were not statistically significant. Regardless of dosage, teat diameter gains in the treatment groups were significantly increased at 90 days about 159%, relative to controls. Cumulative teat diameter gains in the heifers implanted with either 10 mg or 20 mg of estradiol were also significantly increased at 180 days by 100% or 128%, respectively, compared to controls. Teat length gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly increased at 90 days by 200% and 295%, respectively, compared to controls. Cumulative teat length gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly 265% and 325%, respectively, higher than controls. The heifers implanted with either 10 mg or 20 mg of estradiol showed a significant increase in teat volume gains at 90 days of 282% or 246%, respectively, over those of the control heifers. Cumulative teat volume gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly increased at 180 days by 251% and 244%, respectively, compared to control values. However, there were no significant differences m gams of teat diameter, length and volume between heifers implanted with 10 mg and 20 mg of estradiol regardless of 90 days or 180 days. Heifers implanted with 10 mg or 20 mg of estradiol showed the first estrus 20 or 124 days, respectively, faster than controls. Overall, These data strongly suggest that estradiol implantation into heifers stimulates growth and mammary development and hasten the age at which the first estrus in Korean native heifers is observed.

• Korean Journal of Agricultural Science
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• v.19 no.1
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• pp.51-64
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• 1992

This experiment was carried out to elucidate the effects of the thyroid function on lactation in female rats. One hundred and five female rats, whose body weight was approximately 250g with normal parturition, were divided into 3.5 THY, 35 PTU, and 35 CON. The $30{\mu}g$ L-thyroxine per rat was administered subcutaneously for the THY group with 3-days intervals arid 0.03% propylthiouracil solution was drunk for the PTU group. After the treatments body weight, thyroid weights and prolactin levels in serum, and histological changes in thyroid and mammary gland were investigated for 3 weeks with 3-days interval. The results obtained were as follows 1. The body weights of PTU group were lower than those of CON arid THY groups. The changes in body weights were significant between 3 and 6 days and between 15 and 18 days. 2. Differences in thyroid weight among the groups were significant after 9 days, The thyroid weights of PTU group were much higher than those of CON and THY group. 3. The follicular epithelia of PTU group after 6 days showed cuboidal phenomena which were accompanied by hypertrophy and hyperplasia, and this phenomena continued until post weaning period. Those of THY group after 9 days showed a squamous degeneration together with pyknosis. 4. The prolactin concentrations of THY group were higher than the other groups after 12 days. and those of CON group were higher than the others after 18 days. However, those of PTU group were lower all through the period. 5. The secretary epithelial cells of THY and CON groups became cuboidal after 12 days, but after 15 days the differentiation of mammary tissue was progressing in THY group faster than CON group. The degeneration of mammary tissue were observed in PTU group as time lapses, so after 15 days the exfoliation of secretory epithelium and atrophy of alveolus were recognized. 6. The body weights of offspring for all experimental groups were increasing as time lapses, but the values for PTU group were markedly lower than the others.

• The Journal of Korean Obstetrics and Gynecology
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• v.26 no.1
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• pp.1-24
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• 2013

Objectives: The object of this study was to observe the protective effect of Gamijipaesan aqueous extracts(GJS), which has been traditionally used in Korean medicine in obstetrics & gynecological fields as anti-infectious and anti-inflammatory agents, against mastitis induced by Staphylococcus aureus infection in a rat model through antibacterial, antiinflammatory, immunomodulatory, and anti-oxidant effects. Methods: Antibacterial activities of GJS against S. aureus were detected using standard agar microdilution methods, with the effects on the bacterial invasion and intracellular killing of individual test materials in human mammary gland carcinoma cell(MCF-7) and murine macrophages(Raw 264.7) at MIC1/2, MIC and MIC2 concentration levels. In addition, the effects on the cell viability, nitric oxide(NO), tumor necrosis factor(TNF)-${\alpha}$ and interleukin (IL)-6 productions of LPS activated Raw 264.7 cells. The changes on the mammary tissue viable bacterial numbers, myeloperoxidae(MPO), inducible nitric oxide synthetase(iNOS), TNF-${\alpha}$ and IL-6 contents were observed in the S. aureus in vivo intramammary infectious rat model. The anti-bacterial and anti-inflammatory effects were compared with ciprofloxacin and piroxicam, respectively in the present study. Results: MIC of GJS and ciprofloxacin against S. aureus were detected as $0.860{\pm}0.428$ (0.391-1.563) mg/ml and $0.371{\pm}0.262$(0.098-0.782) ${\mu}g/ml$, respectively. In addition, GJS and ciprofloxacin were also showed marked dosage-dependent inhibition of the both bacterial invasion and intracellular killing assays using MCF-7 and Raw 264.7 cells at MIC1/2, MIC and $MIC{\times}2$ concentrations, respectively. $ED_{50}$ against LPS-induced cell viabilities and NO, TNF-${\alpha}$ and IL-6 releases of GJS were detected as 0.72, 0.04, 0.08 and 0.11 mg/ml, and as 19.04, 4.18, 5.37 and 4.27 ${\mu}g/ml$ in piroxicam, respectively. 250 and 500 mg/kg of GJS also inhibit the intramammary bacterial growth, MPO, iNOS, TNF-${\alpha}$ and IL-6 contents in S. aureus in vivo intramammary infected rats, respectively. GJS 500 mg/kg showed quite similar antibacterial and anti-infectious effects as compared with ciprofloxacin 40 mg/kg and also showed similar anti-inflammatory effects as piroxicam 10 mg/kg, in S. aureus in vivo intramammary infectious models. Conclusions: The results obtained in this study suggest that over 250 mg/kg of GJS showed favorable anti-infectious effects against S. aureus infection in a rat model through their antibacterial, anti-inflammatory, immunomodulatory and anti-oxidant effects and therefore expected that GJS can be used as alternative therapies, having both anti-inflammatory and anti-infectious activities. However, more detail mechanism studies should be conducted in future with the efficacy tests of individual herbal composition of GJS and the screening of the biological active compounds in individual herbs. In the present study, GJS 500 mg/kg showed quite similar anti-infectious effects were detected as compared with ciprofloxacin 40 mg/kg treated rats, and also GJS shows quite similar anti-inflammatory effects as compared with piroxicam 10 mg/kg in S. aureus in vivo intramammary infectious rats, but ciprofloxacin did not showed any anti-inflammatory effects, and piroxicam did not showed anti-infectious effects in this study.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.153-158
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• 2008

Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

• The Journal of Korean Obstetrics and Gynecology
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• v.26 no.1
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• pp.25-40
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• 2013

Objectives: The object of this study was to observe the in vitro antibacterial effects of five single(Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix) aqueous herbal extracts, traditionally used for treating various gynecological diseases including mastitis in Korea, against Staphylococcus aureus. Methods: Antibacterial activities against Staphylococcus aureus of aqueous extracts of Pulsatillae Radix, PatrinaeRadix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at Minimal Incubation Concentration(MIC) and $MIC{\times}2$ levels. The effects on the intracellular killing and bacterial invasion of individual test materials were also observed using murine macrophage(Raw 264.7) and human mammary gland carcinoma cell(MCF-7). Results: MIC of aqueous extracts of Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix against Staphylococcus aureus were detected as $0.215{\pm}0.107$ mg/ml, $0.273{\pm}0.107$ mg/ml, $0.469{\pm}0.297$ mg/ml, $11.850{\pm}8.406$ mg/ml, and $0.664{\pm}0.546$ mg/ml, respectively. MIC of Ciprofloxacin was detected as $0.469{\pm}0.297{\mu}g/ml$ at same conditions. In addition, all five single aqueous herbal extracts were also showed marked dosage-dependent inhibition of bacterial growth. The effects of intracellular killing with Raw 264.7 and inhibition of bacterial invasion with MCF-7 cells were detected, in the order of Sophorae Flos, Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix and Sophorae Radix aqueous extracts in the present study. Conclusions: The results obtained in this study suggest that all five single aqueous herbal extracts showed antibacterial effects against Staphylococcus aureus and they also showed dosage-dependent inhibitory effects on the bacterial growth. They showed the significant intracellular killing and inhibition of bacterial invasion effects. It means, all five single aqueous herbal extracts may show potent anti-infectious effects against Staphylococcus aureus for mastitis.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.8
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• pp.1175-1182
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• 2017

Objective: To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases. Methods: TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT). Results: The targeted efficiency for BLG gene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate biallelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.25-34
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• 2003

Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis. To direct hTPO expression in the mammary gland, an expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycin resistance gene (pBT-L neo). Fibroblast cells derived from cow's ear skin tissue were transfected with the expression vector (pBT-L neo) using Lipofectamine. Transfected cells resistant to G418 trea？nt were cultured to form the colonies for more than 2 weeks. The transformed colonies identified by PCR were further expanded prior to nuclear transfer. Reconstructed oocytes with transformed cells were electrofused, activated using calcium ionophore and 6-DMAP, and cultured in vitro for 7 days. Of 35 cell colonies analyzed by PCR, 29 colonies (82.9%) were positive for the hTPO gene. Cleavage and developmental rates to the blastocyst stage of reconstructed embryos with the transformed cells were 65.1% and 23.8%, respectively Of 29 blastocysts that developed from reconstructed embryos with the transformed cells, 27 embryos (93.1%) were transgenic. These results indicate that transgenic bovine embryos can be efficiently produced by somatic cell nuclear transfer using transformed cells.

• Archives of Plastic Surgery
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• v.37 no.5
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• pp.695-698
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• 2010

Purpose: Supernumerary nipple or polythelia is one of the developmental anomalies occurring at the embryonic stage and this anomaly usually arises from the milk line. While this atypical feature is determined during early developmental stage, it may not come out obviously or become troublesome until puberty or lactation. Moreover, sometimes it is confused with a pigmented nevus. Methods: Case 1, a 18-year-old woman with intramammary supernumerary breast consisted of another nipple with middle sized areola on the right lower breast was admitted for a $2.8{\times}3.1\;cm$-sized mass on the right breast which was starting appeared 1 year earlier. The preliminary cytological examination of the material obtained by needle aspiration biopsy from the mass was revealed by fibroadenoma with no malignant change. The patient had the surgical excision of the mass and accessory breast. Case 2, a 16 year-old woman admitted for intra-areolar polythelia of the left breast, even she doesn't have any family history of polythelia. Since she wanted surgical correction of her atypical nipple for aesthetic and psychological reasons, we reconstructed the areola using transposition flaps in an S-plasty design. Results: Case 1, the excised supernumerary nipple showed following histological features. In the superficial layer, an acanthotic and hyperpigmented epithelium with elongated rete ridges was found. In the dermis, there were follicles with hairs surrounded by hypertrophic sebaceous glands. In the deepest portion, abundant secretory glomerules and excretory ducts of apocrine gland type were observed. Case 2, follow-up visits 3 months after the procedure showed a satisfactory result with good shape and projection of the nipple. Conclusion: We report two cases of aberrant mammary tissue who underwent surgical correction, including complete breast (with nipple, areola, and glandular tissue) and intra-areolar polythelia according to the Kajava's classification, and the results were satisfactory.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.1
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• pp.39-46
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• 1985

The fatty acid composition, total number of fatty acid acyl carbon atoms and species of triglycerides from human milk samples obtained during 70 days of lactation from 39 mothers were determined by argentation thin-layer and gas chromatographic procedures. The medium- and long-chain saturated fatty acids(8:0, 1O:0, 12:0 and 14:0) which are formed exclusively by synthesis within mammary gland increased significantly from colostrum to mature milk. Long-chain saturated fatty acids(16 : 0, 22 : 0 and 24: 0) were significantly higher than tile levels found in transitional and mature milk. The precursors of w C- and w 3-series, 18:2 w 6 and 18:3 w 3, were increased slightly in progressing lactation. Colostrum contained significantly higher proportions of 18:1 w 9 and w 6- and w 3- derived long-chain polyunsaturated fatty acids than transitional milk, and these levels were further reduced in mature milk. The triglycerides of human milk lipids which were made up of 30-60 acyl carbon atoms showed a pattern with major contributions made by the glycerides with 44-52 acyl carbon atoms. The levels of triglycerides with less than 46 acyl carbon atoms increased significantly with the elapse of lactation period, whereas those with more than 50 acyl carbon atoms decreased significantly. The fully saturated trig1ycerides increased significantly as the lactation proceeded, but the dienoic triglycerides declined significantly.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.