• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.60-60
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• 2001

Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis by influencing on the development and maturation of megakaryocyte and platelet production. To induce hTPO production in the mammary gland, expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycine resistance gene for transfection into fibroblasts. Bovine fibroblast cells derived from female ear skin were transfercted with the expression vector using Lipofectamine (Life Technology, NY). Transected cells resistant to G4l8 treatment (600 $\mu\textrm{g}$/$m\ell$) were recovered and colony formation was initiated at 13 days. The colonies with about 1 cm diameter were picked and analysed by PCR. Single transfected cells were individually transferred to enucleated oocytes. After electrofusion, the reconstructed embryos were exposed to calcium ionophore (5uM) for 5 min followed by treatment with 6-DMAP (2.5 mM) for 4h. The nuclear transfer embryos were cultured in CRlaa medium at 38.5C, 5% $CO_2$ for 7 days. Twenty three of 29 (79.3%) colonies were proved to be hTPO transfectants by PCR. The colonies were further passaged and used to produce transgenic embryos using nuclear transfer. Cleavage and developmental rates of reconstructed embryos to the blastocyst stage were 65.1% and 39.4%, respectively Of 22 blastocysts that developed from reconstructed embryos with the transfected cell, 20 embryos (90.9%) were positive for hTPO by using PCR analysis. The results suggest that somatic cell nuclear transfer is efficient for production of transgenic embryos.

• Genomics & Informatics
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• v.21 no.2
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• pp.19.1-19.13
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• 2023

Liver cancer, particularly hepatocellular carcinoma (HCC), poses a significant global threat to human lives. To advance the development of innovative diagnostic and treatment approaches, it is essential to examine the hidden features of HCC, particularly its 3D genome architecture, which is not well understood. In this study, we investigated the 3D genome organization of four HCC cell lines-Hep3B, Huh1, Huh7, and SNU449-using in situ Hi-C and assay for transposase-accessible chromatin sequencing. Our findings revealed that HCC cell lines had more long-range interactions, both intra-and interchromosomal, compared to human mammary epithelial cells (HMECs). Unexpectedly, HCC cell lines displayed cell line-specific compartmental modifications at the megabase (Mb) scale, which could potentially be leveraged in determining HCC subtypes. At the sub-Mb scale, we observed decreases in intra-TAD (topologically associated domain) interactions and chromatin loops in HCC cell lines compared to HMECs. Lastly, we discovered a correlation between gene expression and the 3D chromatin architecture of SLC8A1, which encodes a sodium-calcium antiporter whose modulation is known to induce apoptosis by comparison between HCC cell lines and HMECs. Our findings suggest that HCC cell lines have a distinct 3D genome organization that is different from those of normal and other cancer cells based on the analysis of compartments, TADs, and chromatin loops. Overall, we take this as evidence that genome organization plays a crucial role in cancer phenotype determination. Further exploration of epigenetics in HCC will help us to better understand specific gene regulation mechanisms and uncover novel targets for cancer treatment.

• Journal of Veterinary Science
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• v.24 no.5
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• pp.72.1-72.16
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• 2023

Background: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the surface of Streptococcus dysgalactiae, coded with gapC, is a glycolytic enzyme that was reported to be a moonlighting protein and virulence factor. Objective: This study assessed GAPDH as a potential immunization candidate protein to prevent streptococcus infections. Methods: Mice were vaccinated subcutaneously with recombinant GAPDH and challenged with S. dysgalactiae in vivo. They were then evaluated using histological methods. rGAPDH of mouse bone marrow-derived dendritic cells (BMDCs) was evaluated using immunoblotting, reverse transcription quantitative polymerase chain reaction, and enzyme-linked immunosorbent assay methods. Results: Vaccination with rGAPDH improved the survival rates and decreased the bacterial burdens in the mammary glands compared to the control group. The mechanism by which rGAPDH vaccination protects against S. dysgalactiae was investigated. In vitro experiments showed that rGAPDH boosted the generation of interleukin-10 and tumor necrosis factor-α. Treatment of BMDCs with TAK-242, a toll-like receptor 4 inhibitor, or C29, a toll-like receptor 2 inhibitor, reduced cytokines substantially, suggesting that rGAPDH may be a potential ligand for both TLR2 and TLR4. Subsequent investigations showed that rGAPDH may activate the phosphorylation of MAPKs and nuclear factor-κB. Conclusions: GAPDH is a promising immunization candidate protein for targeting virulence and enhancing immune-mediated protection. Further investigations are warranted to understand the mechanisms underlying the activation of BMDCs by rGAPDH in a TLR2- and TLR4-dependent manner and the regulation of inflammatory cytokines contributing to mastitis pathogenesis.

• Korean Journal of Veterinary Research
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• v.63 no.4
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• pp.34.1-34.9
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• 2023

Dogs exhibit patterns of health issues that vary by life stage. An understanding of disease incidence with respect to breed and age/life stage could be an important component of canine health management and welfare. This study aimed to describe the age-specific disease incidence of 3 small dog breeds that attended veterinary clinics in the Republic of Korea, based on data from electronic veterinary medical records (EVMRs). A total of 40,785 EVMRs from Maltese (n = 21,355), Miniature Poodle (n = 11,658) and Shih Tzu dogs (n = 7,772) were analyzed. Common health problems in 3 small dog breeds were 'diseases of the skin' and 'diseases of the ear,' respectively. Among dogs aged ≤ 3 years, 'preventive medicine' was the most common cause cited for veterinary clinic visits. Among dogs aged 4 to 8 years, the most frequent health problems were 'diseases of the skin.' Among dogs aged 9 to 13 years, 'heart diseases,' 'kidney diseases,' 'mammary gland tumor,' and 'neoplasia (unspecified)' were considerably more frequent, compared to the rates in dogs ≤3 years. Among dogs aged ≥ 14 years, 'heart diseases' and 'sneezing/cough' were the main health problems. In all breeds, the frequencies of 'diseases of circulatory system,' 'diseases of respiratory system,' 'diseases of the nervous system,' 'endocrine' and 'neoplasia' increased rapidly with aging. This surveillance could inform strategies for disease screening tests and management based on life stage in these dog breeds and enable more effective health management.

• Journal of the Korean Society of Radiology
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• v.83 no.3
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• pp.687-692
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• 2022

Male breast cancer is rare, accounting for approximately 1% of breast cancers. Metastasis from extra-mammary malignancy to the breast in men is extremely rare. The most common primary tumors that metastasize to the breast in male are prostate, lung, stomach, colorectal cancer, melanoma, and sarcoma. To our knowledge, only a few cases of bladder cancer presenting with metastasis to the male breast have been reported, and metastasis with infiltration rather than mass is extremely rare. We report imaging findings on mammography and ultrasonography in a 59-year-old male with bladder cancer metastatic to the breast.

• IMMUNE NETWORK
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• v.20 no.4
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• pp.33.1-33.19
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• 2020

We tested how adjuvants effect in a cancer vaccine model using an epitope derived from an autoactivation loop of membrane-type protease serine protease 14 (PRSS14; loop metavaccine) in mouse mammary tumor virus (MMTV)-polyoma middle tumor-antigen (PyMT) system and in 2 other orthotopic mouse systems. Earlier, we reported that loop metavaccine effectively prevented progression and metastasis regardless of adjuvant types and TH types of hosts in tail-vein injection systems. However, the loop metavaccine with Freund's complete adjuvant (CFA) reduced cancer progression and metastasis while that with alum, to our surprise, were adversely affected in 3 tumor bearing mouse models. The amounts of loop peptide specific antibodies inversely correlated with tumor burden and metastasis, meanwhile both T_H1 and T_H2 isotypes were present regardless of host type and adjuvant. Tumor infiltrating myeloid cells such as eosinophil, monocyte, and neutrophil were asymmetrically distributed among 2 adjuvant groups with loop metavaccine. Systemic expression profiling using the lymph nodes of the differentially immunized MMTV-PyMT mouse revealed that adjuvant types, as well as loop metavaccine can change the immune signatures. Specifically, loop metavaccine itself induces T_H2 and T_H17 responses but reduces T_H1 and T_reg responses regardless of adjuvant type, whereas CFA but not alum increased follicular T_H response. Among the myeloid signatures, eosinophil was most distinct between CFA and alum. Survival analysis of breast cancer patients showed that eosinophil chemokines can be useful prognostic factors in PRSS14 positive patients. Based on these observations, we concluded that multiple immune parameters are to be considered when applying a vaccine strategy to cancer patients.

• 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.

• 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 Chest Surgery
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• v.38 no.8 s.253
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• pp.551-556
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• 2005

Background: Complete arterial off-pump coronary artery bypass grafting (OPCAB) by sequential anastomoses with one or two arterial grafts provides favorable outcomes. However, problems of insufficient graft length, hypopefusion, kinking of graft, and unfavorable course of graft may be encountered. To solve these problems, we have used different technique with multiple arterial Y composite graft to allow end-to-side rather than sequential anastomoses and evaluated the results of this method. Material and Method: Between February 2003 and October 2004, If patients underwent total arterial OPCAB using multiple arterial V composite grafts with left internal mammary artery (LIMA), radial artery (RA), and right internal mammary artery (RIMA). We divided RA into multiple segments by number of distal target site after measuring of individual proper length and constructed arterial composite graft. One of segments was sutured end-to-side to LIMA and other segment was sutured end-to-side to the previously constructed radial graft. Postoperative graft patency was evaluated in 6f patients by multi-slice computed tomegraphy. Result: An average of $2.5\pm0.6$ arteries and $3.7pm0.7$ distal anastomoses per patient were done. There was no perioperative myocardial infarction, clinical hypoperfusion syndromes, and operative mortality. Postoperative mean CK-MB level was $17.4pm29.7\;IU/L.$Overall graft patency was $99.1\%\;(214/216)(LIMA:\;100\%,\;RA:\;98.4\%,\;RIMA:\;100\%).$ Conclusion: This technique allows total arterial OPCAB without technical problems and provides excellent early clinical results and graft patency. We believe that this technique is more convenient in the obtuse marginal area compared to sequential technique, and helpful in patients who require complex arterial grafting.

• Investigative Magnetic Resonance Imaging
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• v.7 no.1
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• pp.22-30
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• 2003

Purpose : The efficacy of magnetic resonance imaging for evaluating coronary artery disease has been reported. In this study, we evaluated the usefulness of breath-hold segmented K-space cine MR imaging for evaluating the patency of coronary artery bypass grafts (CABG). Materials and Method s : Thirty eight patients with a total of 92 CABGs (36 internal thoracic arteries and 56 saphenous vein grafts) were evaluated using segmented K-space cardiac-gated fast gradient echo sequence (2D-FASTCARD) MR imaging. MR magnitude images were evaluated from the hard copies by two independent observers. A graft was defined as patent if it was seen as a bright small round area on at least two consecutive images throughout the cardiac cycle at a position consistent with the expected location for that graft. Results : MR images were obtained successfully for 23 patients (61%). The sagittal planes were most helpful in visualizing the cross-section of sapheneous vein bypass graft to left circumflex artery branch, whereas the transverse planes were used for identification of internal mammary artery grafts to left anterior descending coronary artery or its branch and identification of saphenous vein grafts to right coronary artery. Forty five grafts were visible using this MR technique, while the grafts were not visible on seven saphenous vein grafts and two internal mammary artery grafts. In two patients showing symptoms of myocardial ischemia, one or two bypass grafts were not visible. Imaging, perpendicular plane to a CABG was important to visualize the flow inside the CABG with maximum sensitivity. Conclusion : Evaluation of patency of the bypass graft was clinically feasible by 2D-FASTCARD MR imaging, whereas any invisible bypass grafts should be further studied by contrast-enhanced MR angiography or by conventional angiography for confirmation of abnormalities.