• Development and Reproduction
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• v.11 no.3
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• pp.253-261
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• 2007

Lactadherin is a glycoprotein of human milk fat globule membrane that binds to mucin and butyrophilin forming the protein complex. Especially, mucin and lactadherin in human milk efficiently protect infants with poor immune functions right after birth from infections by microorganisms and play important roles for their early survival, growth and development. Lactadherin inhibits the propagation and growth of rotavirus that is a global pathogen causing infants' diarrhea. Recently this protein was known to promote neovascularization and its deficiency related to develop Alzheimer's disease. In this study, the basic biochemical and physiological aspects of lactadherin were investigated. Messenger RNAs were isolated from mammary tissues from Korean women patients to clone a 1.2 kb cDNA and sequenced its DNA to determine its amino acid sequences. The cDNA was cloned to express its 43 kD protein in E. coli, which was confirmed by Western blot. The recombinant protein was purified and injected to 2 rabbits to raise antibodies against it. The semi-purified milk fat globule membrane proteins from Korean women was analyzed by Western blot using the rabbit antibody to give 70, 55, 46, 30 kD bands. Also several polymorphism and SNPs of lactadherin gene from Korean women were observed compared with those of Caucasian women.

• 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.

• Journal of Veterinary Clinics
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• v.27 no.3
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• pp.252-256
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• 2010

Bovine mastitis, defined as an inflammation of the mammary gland, is usually associated with bacterial infection. Thus, treatment and control of mastitis relies primarily on antimicrobial therapy. This study investigated the bactericidal actions of silver ion against causing various bovine mastitis pathogens. Minimal inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs) and bactericidal activity times and concentrations of silver ion against pathogens were determined. The effect of silver ion on bacterial morphology was studied by scanning electron microscopy (SEM). The MICs and MBCs of silver ion for various bacteria strains ranged from 1.9-15.6 ${\mu}g$/ml. SEM images indicated formation of a pit, distortion and disruption of cell walls in silver treated Staphylococcus aureus and Escherichia coli. The results demonstrate that silver ion has a bactericidal capacity against causing various pathogens of bovine mastitis and suggest that silver ions may be exploitable as a therapeutic/preventative tool of bovine mastitis.

• Journal of Biomedical Engineering Research
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• v.33 no.2
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• pp.89-97
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• 2012

The purpose of this study is to develop the guideline of the physicochemical and mechanical properties evaluation for silicone gel filled breast implants. First of all, the use and development status for silicone gel filled breast implants were investigated, and then, standard and criteria about performance evaluation established by the international organizations such as ASTM, FDA guidance and ISO were examined. To evaluate the mechanical properties, data research and testing for breaking strength, elongation, tensile set, joint intensity, silicone gel cohesion, weight loss from heating, static rupture resistance, impact resistance test, fatigue test, and gel bleed were performed. On the other hand, to evaluate the physicochemical properties, volatile matter, extent of cross linking, heavy metals, and extractable were analyzed. In this study, results for general function, mechanical properties and physicochemical properties were examined and reviewed for the accordance with international standard, and objective and standardized guideline was provided.

• Parasites, Hosts and Diseases
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• v.55 no.6
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• pp.601-606
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• 2017

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.

• Development and Reproduction
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• v.14 no.1
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• pp.13-19
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• 2010

We developed a novel dicistronic system for the expression of target cDNA sequences in the milk of transgenic animals using goat beta-casein/hGH fusion construct, pGbc5.5hGH (Lee, 2006) and internal ribosome entry site (IRES) sequences of encephalomyocarditis virus (EMCV). Granulocyte colony-stimulating factor (hG-CSF) cDNA was linked to 3' untranslated region of hGH gene in the pGbc5.5hGH via EMCV IRES sequences. Transgenic mice were generated by microinjection and transgene expression was examined in the milk and mammary gland of transgenic mice at 10 days of lactation. Northern blot analysis showed that hGH gene and hG-CSF cDNA were transcribed as a single dicistronic mRNA. The hG-CSF and hGH proteins were independently translated from the dicistronic mRNA and secreted into the milk of transgenic mice. The highest concentration of hG-CSF and hGH in the milk of transgenic mice were $237{\mu}g/m{\ell}$ and $8,990{\mu}g/m{\ell}$, respectively. In contrast, another hG-CSF expression cassette, in which hG-CSF genomic sequences were inserted into a commercial milk-specific expression vector (pBC1), generated a lower level ($91{\mu}g/m{\ell}$) of hG-CSF expression in the milk of transgenic mice. These results demonstrated that the novel pGbc5.5hGH-based dicistronic construct could be useful for an efficient cDNA expression in the milk of transgenic animals.

• BMB Reports
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• v.35 no.4
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• pp.371-376
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• 2002

The receptor activator of nuclear factor kappa B (RANK) is a member of the tumor necrosis factor (TNF) receptor superfamily. It plays a critical role in osteoclast differentiaion, lymph node organogenesis, and mammary gland development. The stimulation of RANK causes the activation of transcription factors NF-${\kappa}B$ and activator protein 1 (AP1), and the mitogen activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK). In the signal transduction of RANK, the recruitment of the adaptor molecules, TNF receptor-associated factors (TRAFs), is and initial cytoplasmic event. Recently, the association of the MAPK kinase kinase, transforming growth factor-$\beta$-activated kinase 1 (TAK1), with TRAF6 was shown to mediate the IL-1 signaling to NF-${\kappa}B$ and JNK. We investigated whether or not TAK1 plays a role in RANK signaling. A dominant-negative form of TAK1 was discovered to abolish the RANK-induced activation of AP1 and JNK. The AP1 activation by TRAF2, TRAF5, and TRAF6 was also greatly suppressed by the dominant-negative TAK1. the inhibitory effect of the TAK1 mutant on RANK-and TRAF-induced NF-${\kappa}B$ activation was also observed, but less efficiently. Our findings indicate that TAK1 is involved in the MAPK cascade and NF-${\kappa}B$ pathway that is activated by RANK.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.5937-5946
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• 2012

The term endocrine disruptors is used to describe a variety of natural and manmade substances that have the capacity to potentially interfere with and modify the normal physiology of endocrine system either by mimicking, blocking or modulating the actions of natural endogenous hormones. The rising incidence of breast cancer over the last 50 years and the documented higher incidence in urban as compared to rural areas suggest a relationship to the introduction and increased use of xenoestrogens in our environment. The literature has developed over the last decades where initial experiments on endocrine disruptors did not support an involvement in breast cancer, and then evidence mounted implicating various environmental factors including hormones, endocrine disrupting chemicals and non-endocrine disrupting environmental carcinogens in the pathogenesis of breast cancer. Available data support the hypothesis that exposure to endocrine disruptors in utero leaves a signature on mammary gland morphogenesis so that the resulting dysgenic gland becomes more predisposed to develop tumors upon exposures to additional insults later on during life. Exceptionally, exposure to phytoestrogens could be beneficial to human health. Most of the available data are from well developed countries while the developing countries are still understudied regarding these issues. Here, we raise a note of caution about potential role of environmental toxins including endocrine disruptors in breast cancer development and call for serious measures to be taken by all involved parties in the developing world.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.12
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• pp.5159-5164
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• 2016

Objective: The dynamics of morbidity, disability and death rates due to malignant neoplasms in the population in Uralsk city of the Republic of Kazakhstan were studied for 2011-2015, with a focus on age and sex, as well as tumor location. Methods: Statistics for total morbidity, primary disability and mortality from cancer in the adult population of the city of Uralsk for 2011-2015 were calculated per 100 thousand. Estimation of morbidity was based on data from form - $N{\underline{o}}12$ ${\ll}$Report on the number of diseases registered in patients living in the area of health care organizations and patient population under medical observation". Evaluation of primary disability was based on form $N{\underline{o}}7$ ${\ll}$The distribution of newly recognized disabled by disease class, age, sex and disability groups" for 2011-2015 in Ural city and analysis of cancer was carried out using annual form 7 "Report on the sick, and diseases of malignant neoplasms". Result: The most common localizations of cancer were the trachea, bronchi, lungs, stomach and mammary glands. High death rates were noted for patients with cancer of the trachea, bronchi, lung, as compared to stomach and esophagus. Conclusion: The results of our investigation and data in the literature indicate that regional characteristics influence the impact of risk factors associated with cancer. An unfavorable environmental background contributes to ill health of urban populations, contributing to development of cancer. Moreover behavioral risk factors are very important, such as smoking, alcohol drinking, and an unhealthy diet. All these factors require urgent adoption of a package of measures for prevention, early detection and timely treatment. Detailed study of cancer is necessary to develop national programs and activities for prevention and control.

• Journal of the Korean Society of Visualization
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• v.13 no.1
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• pp.43-48
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• 2015

Collective cell migration is a fundamental phenomenon observed in various biological processes such as development, wound healing, and cancer metastasis. During the collective migration, cells undergo changes in their phenotypes from those of stable to the migratory state via the process called epithelial-mesenchymal transition (EMT). Recent findings in biology and biochemistry have shown that EMT is closely related to the cancer invasion or metastasis, but not much of the correlations in kinematics and physical forces between the neighboring cells are known yet. In this study, we aim to understand the cell migration and stress distribution within the expanding cell cluster. We constructed the in vitro cell cluster on the hydrogel, employed traction force microscopy (TFM) and monolayer stress microscopy (MSM) to visualize the physical forces within the expanding cell monolayer. During the expansion, cells at the cluster edge exhibited enhanced motility and developed focal adhesions that are the essential features of EMT while cells at the core of the cluster maintained the epithelial characteristics. In the aspect of mechanical stress, the cluster edge had the highest traction force of ~90 Pa directed toward the cluster core, which means that cells at the edge actively pull the substrate to make the cluster expansion. The cluster core of the tightly confined cells by neighboring cells had a lower traction force value (~60 Pa) but the highest intercellular normal stress of ~800 Pa because of the accumulation of traction from the edge of the monolayer.