• Title/Summary/Keyword: mammary

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Transmission of Bovine $\beta-Casein/Human$ Lactoferrin Fusion Gene in Transgenic Cattle

  • Han Yong-Mahn;Koo Deog-Bon;Park Jung-Sun;Kim Young-Hun;Lee Kea-Joung;Lee Kyung-Kwang
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.235-239
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    • 2005
  • This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

Breast Reconstruction with Superior Gluteal Artery Perforator Flap in Asian (동양인에서 위볼기동맥 천공지판을 이용한 유방재건)

  • Jeong, Woo Shik;Lee, Taek Jong;Eom, Jin Sup
    • Archives of Reconstructive Microsurgery
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    • v.22 no.1
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    • pp.7-12
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    • 2013
  • Purpose: Breast reconstruction with lower abdominal tissue can produce the best outcome with acceptable rates of long-term complication. However, for cases in which sufficient abdominal tissue is not available, an superior gluteal artery perforator (SGAP) flap can be considered as the next option for autologous breast reconstruction. Materials and Methods: Among a total of 63 women who underwent breast reconstruction with free autologous tissue transfer from July 2010 to April 2011, SGAP flap was performed for four patients. In two cases, patients did not have enough abdominal tissue for sizable breast reconstruction. In another case, the patient had a long abdominal scar due to donor hepatectomy of liver transplantation. In the last case, which was a revisional case after radiation necrosis of a previous pedicled transverse rectus abdominis musculocutaneous (TRAM) flap, a large amount of healthy skin and soft tissue was needed. SGAP flap was elevated in lateral decubitus position. The internal mammary vessels were used for recipient vessels in all cases. Results: Breast reconstruction was performed successfully in all four cases without flap loss. Donor site complication was not observed, except for one case of seroma. The shape of the reconstructed breast was satisfactory in all patients. Conclusion: SGAP flap is an excellent alternative option for the TRAM or deep inferior epigastric artery perforator flap for breast reconstruction. In terms of narrower width, harder consistency of soft tissue, and shorter pedicle, it is clear that the SGAP flap is less competent than the TRAM flap. However, in cases where abdominal tissue is not available, SGAP flap is the only way of providing a large amount of healthy tissue.

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Changes in Reproductive Function and White Blood Cell Proliferation Induced in Mice by Injection of a Prolactin-expressing Plasmid into Muscle

  • Lee, Jung-Sun;Yun, Bo-Young;Kim, Sang-Soo;Cho, Chunghee;Yoon, Yong-Dal;Cho, Byung-Nam
    • Molecules and Cells
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    • v.22 no.2
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    • pp.189-197
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    • 2006
  • Prolactin (PRL) is a pituitary hormone involved in various physiological processes, including lactation, mammary development, and immune function. To further investigate the in vivo and comparative endocrine roles of PRL, mouse PRL cDNA fused to the cytomegalovirus promoter, was introduced into muscle by direct injection. Previously we studied the function of rat PRL using the same protocol. PRL mRNA was detected in the muscle following injection by RT-PCR and subsequent Southern blot analysis. PRL was also detected and Western blot analysis revealed a relatively high level of serum PRL. In the pCMV-mPRL-injected female mice, the estrous cycle was extended, especially in diestrus stage and the uterus thickening that was shown in normal estrous stage was not observed. In the pCMV-mPRL-injected male mice, new blood vessels were first found at 5 weeks of age and fully developed blood vessels were found after 8 weeks in the testis. The number of Leydig cells increased within the testis and the testosterone level in serum was observed high. Finally, the number of white blood cells (WBCs) increased in the pCMV-mPRL-injected mice. The augmentation of WBCs persisted for at least 20 days after injection. When injection was combined with adrenalectomy, there was an even greater increase in number of WBCs, especially lymphocytes. This increase was returned normal by treatment with dexamethansone. Taken together, our data reveal that intramuscularly expressed mouse PRL influences reproductive functions in female, induces formation of new blood vessels in the testis, and augments WBC numbers. Of notice is that the Leydig cell proliferation with increased testosterone was conspicuously observed in the pCMV-mPRL-injected mice. These results also suggest subtle difference in function of PRL between mouse and rat species.

The Experimental Study on anti-inflammatory Effects of Kagamsunbanghwalmyungeum(KSBH) (가감선방활명음(加減仙方活命飮)의 항염작용(抗炎作用)에 대한 실험적(實驗的) 연구(硏究))

  • Shin, Sun-Mi;Kim, Soo-Min;Kim, Eui-Il;Lee, Jung-Eun;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.1
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    • pp.125-143
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    • 2007
  • Purpose : This study was to investigate the anti-inflammatory effects of Kagamsunbanghwalmyungeum(KSBH) which has been medicated the patient such as breast abscess. Methods : In the study of anti-inflammatory effects, KSBH was investigated using cultured cells and a murine models. As for the parameters of inflammation, level of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were determined in mouse lung fibroblast cell(mLFC) and RAW 264.7 cells. Results : The cytotoxicity on mouse lung fibroblast cells(mLFC) was not served at all concentration of KSBH. KSBH in RAW264.7 cell inhibited IL-l${\beta}$, IL-6, TNF-${\alpha}$, COX-2 and NOS-II mRNA genes expression in a concentration-dependent manner, and specially inhibited NOS-II production very significantly at 100${\mu}$g/ ml. KSBH inhibited NO production significantly in a concentration-dependent manner and ROS production in a concentration-dependent manner. KSBH inhibited IL-l${\beta}$, IL-6 and TNF-${\alpha}$ production significantly in serum of acute anti-inflammation-induced mice. KSBH increased the survival rate from the 3rd day on LPS-induced lethal endotoxemia. Conclusion : These results suggest that Kagamsunbanghwalmyungeum(KSBH) can be useful in treating a lot of women mammary diseases caused by inflammation such as breast abscess.

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Isolation of Brucella spp from sere-positive native bulls and calves below twelve months old (브루셀라병 혈청검사 양성 수소와 12개월령 이하 소에서의 균 분리 및 동정)

  • Ryu, Jae-Yun;Bun, Jung-Hyun;Lee, Hee-Young;Lee, Yong-Chang;Lee, Jong-Jin;Song, Young-Gak;Nam, Hyang-Mi
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.375-384
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    • 2007
  • This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.

Analysis of premature death of Sprague-Dawley rats in carcinogenicity studies

  • Son, Woo-Chan;Kim, Bae-Hwan
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.373-378
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    • 2004
  • To help the interpretation of causes of death, it is critical that the background incidence of factors contributing to death be recorded and archived. Information was gathered from the control groups of 19 rat carcinogenicity studies. All cases of death occurring within the 2-year period were reviewed. Out of 1124 males and 1084 females, 720 male (64.1%) and 689 female (63.6%) decedents were recorded. There was no difference in the probability of survival between two sexes. Analysis of factors contributing to death revealed that 400 males (48.7%) had neoplastic changes, 189 males (23.0%) had non-neoplastic lesions, and 232 males (28.3%) died from unknown causes. In females, these figures were 627 (76.4%), 62 (7.6%) and 132 (16.0%), for neoplastic, non-neoplastic and unknown findings, respectively. It could be suggested that the risk of death by non-neoplastic reasons was higher in the males than in the females, whereas females were more likely to be affected by tumours. In the neoplastic causes of death, pituitary tumours were the most common in both sexes, followed by mammary tumours in females, and haemopoietic tumours in males. In non-neoplastic cause of death, renal diseases were the most common in both sexes, followed by skin diseases and cardiovascular diseases in males, and skin diseases and poditis in males. A relatively large number of animals (28.3% in males and 16.0% in females) were found dead, without any significant clinical or histologically identifiable cause. Most of the animals with pituitary tumours were killed in extremis and the proportion of females (70.1%) being greater than males (46.8%). There were no case which died by accident, and also only minimal incidence which died by bleeding procedures.

Cell Survival, Apoptosis and AMPK-COX-2 Signaling Pathway of Mammary Tumor Cells after Genistein Treatment Combined with Estrogen

  • Lee, Yun-Kyoung;Hwang, Jin-Taek;Kim, Young-Min;Park, Ock-Jin
    • Preventive Nutrition and Food Science
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    • v.12 no.4
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    • pp.197-201
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    • 2007
  • Genistein is an active component of legumes and other related food shown to be associated with prevention of degenerative diseases such as cancer through inducing signaling pathways. Treatment of genistein resulted in the induction of apoptosis in the cultured cancer cells. This induction of apoptosis was demonstrated by the Tunel assay in these cells. Unveiling the potential of genistein in cytotoxicity via apoptosis when it is treated with estrogen can predict the therapeutic capability of genistein in breast cancers in the presence of endogenous estrogen. We have found that apoptosis induced by genistein treatment in the presence of estrogen is agonistic or antagonistic depending on the concentrations and treatment periods applied in MCF-7 breast cancer cells. For the suppression of cell survival, 24 hr of treatment was required to induce a synergistic agonistic response between estrogen and genistein at low concentrations of genistein. After this period, the agonistic pattern of genistein to estrogen disappeared. The decrement of COX-2 expression in MCF-7 cells treated with genistein was accompanied with the activation of AMPK only at a high concentration of genistein. This association between AMPK activation and down-regulation of COX-2 by genistein was dampened in the presence of estrogen. It was also demonstrated that genistein and estrogen regulate cell survival and apoptosis by modulating p53 and caspase-3 in the opposite direction. These results suggest that genistein has the potential to control breast cancer development, and co-treatment with estrogen can cause agonistic or antagonistic action on breast cancer cell control.

Expression of Human Lactoferricin in HC11 Cells (HC11 세포에서 인체 락토페리신의 발현)

  • Nam, Myoung-Soo
    • Korean Journal of Agricultural Science
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    • v.28 no.2
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    • pp.92-98
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    • 2001
  • Lactofenicin is an antibacterial peptide fragment (about 5 kD) derived from lactoferrin (80 kD) that displays the various biological functions. The production of a human lactoferricin (Lactoferricin H) in mouse HC11 mammary epithelial cells was achieved by placing its cDNA under the control of the bovine ${\beta}$-casein gene. To express lactoferricin H in this cell culture system, constructed a hybride-splice signal consisting of bovine ${\beta}$-casein intron I and rabbit ${\beta}$-globin intron II, and a DNA fragment spanning intron 8 of the bovine ${\beta}$-casein gene. Expression of lactofenicin H from this expression vector was identified by RT-PCR, northern and dot blot analysis. RT-PCR using total RNA of HC11 cells transfected with pBL1-cin expression vector yielded a product identified as having a size of the 150bp. Northern blot analysis was identified about 2.3 kb. In dot blot analysis, recombinant lactofenicin H was recognized with anti-human lactofrrnin polyclonal antibody.

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Synthesis and Antitumor Evaluation of Acyclic 1-[${\omega}$-(N^I-2-chloroethyl-N^I-nitrosoureido)alkyl]thymidine Nucleoside Analogues

  • Kim, Jack-C.;Kim, Young-Hyun;Park, Jin-Il;Kim, Seon-Hee;Choi, Soon-Kyu
    • Archives of Pharmacal Research
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    • v.20 no.3
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    • pp.259-263
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    • 1997
  • In the preparation of acyclic thymidine nucleoside analogues, $K_2CO_3$(or NaH) treated thymine in DMSO was alkylated with .omega.-chloroalkyl nitrite (Cl-(CH_2)n-CN; n=1, 2, 3, 4) to provide an isomeric mixture of 1-(${\omega}$-cyanoalkyl)thymine (2a-d) and 1,3-bis(${\omega}$-cyanoalkyl)thymine in approximately 5:1 ratios. Reduction of the cyano function 2a-d with $NaBH_{4}/CoCl_{2}$ center dot$ 6H_{2}O$gave the corresponding 1-(${\omega}$aminoalkyl)thymine (3a-d). The newly formed primary amino function in 3a-d was directly reacted with 2-chloroethylisocyanate to afford the 1-[.omega.($N^{I}$-2-chloroethylureido) alkyl]thymine (4a-d) in good yields. Nitrosation of 1-[5-($ N^{I}-2$-chloroethylureido)pentyl] thymine (4d) with glacial acetic acid and dry $NaNO_{2}$-powder in anhydrous $CH_{2}Cl_{2}$gave two types of regioisomeric nitrosoureas, 1-[5-($N^{I}$--chloroethyl-$N^{I}$--nitrosoureido)pentylithymine (5d) and 1-[5-($N^{I}-2$-chloroethyl-N-nitrosoureido)pentyllthymine in approximately 5 :1 ratios. The in vitro cytotoxicity of the synthesized compounds (2a-d, 3a-d, 4a-d and 5a-d) against three cell lines (K-562, P-388 and FM-3A) are measured as $IC^{50}$ values. Compounds 3b and 4c showed moderate activities against all three cell lines, and all other compounds were found to be not active.

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Synthesis and antitumor evaluation of $\alphamethylene-\gamma-butyrolactone-linked$ to 5-substituted uracil nucleic acid bases

  • Kim, Jack-C.;Kim, Ji-A;Kim, Si-Hwan;Park, Jin-Il;Kim, Seon-Hee;Park, Soon-Kyu;Park, Won-Woo
    • Archives of Pharmacal Research
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    • v.19 no.3
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    • pp.235-239
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    • 1996
  • Six, heretofore undescribed, $5^I-Methyl-5^I-(5-Substituted uracil-1-ylmethyl)-2^I-oxo-3^I-methylenetetrahydrofurans(F, Cl, Br, l, CH_3, H)(6a-f)$were synthesized and evaluated against three cell lines (FM-3A, P-388 and U-937). For the preparation of .alpha.-methylene-.gamma.-butyrolactone bearing 5-substituted uracils (6a-f), the effcient Reformatsky type reaction was employed which involves the treatment of ethyl .alpha.(bromomethyl) acrylate and zinc with the respective 5-substituted uracil-1-ylacetones (5a-f). The acetone derivatives (5a-f) were directly obtained by the respective alkylation reaction of 5-substituted uracils with chloroacetone in the presence of $K_{2}$$CO_{3}$(or NaH). These lactone compounds 6a-f exhibited moderate to significant activity in all of the three cell lines, and 6b, 6c and 6e showed significant antitumor activities (inhibitory concentrations ($IC_{50}$) ranged from 1.3-3.8 .mu.g/ml.

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