• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1394-1406
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• 2015

The AT motif-binding factor (ATBF1) not only interacts with protein inhibitor of activated signal transducer and activator of transcription 3 (STAT3) (PIAS3) to suppress STAT3 signaling regulating embryo early development and cell differentiation, but is required for early activation of the pituitary specific transcription factor 1 (Pit1) gene (also known as POU1F1) critically affecting mammalian growth and development. The goal of this study was to detect novel nucleotide variations and haplotypes structure of the ATBF1 gene, as well as to test their associations with growth-related traits in goats. Herein, a total of seven novel single nucleotide polymorphisms (SNPs) (SNP 1-7) within this gene were found in two well-known Chinese native goat breeds. Haplotypes structure analysis demonstrated that there were four haplotypes in Hainan black goat while seventeen haplotypes in Xinong Saanen dairy goat, and both breeds only shared one haplotype (hap1). Association testing revealed that the SNP2, SNP5, SNP6, and SNP7 loci were also found to significantly associate with growth-related traits in goats, respectively. Moreover, one diplotype in Xinong Saanen dairy goats significantly linked to growth related traits. These preliminary findings not only would extend the spectrum of genetic variations of the goat ATBF1 gene, but also would contribute to implementing marker-assisted selection in genetics and breeding in goats.

• Journal of Plant Biotechnology
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• v.35 no.4
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• pp.281-290
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• 2008

In plant, Receptor-like kinases (RLKs) are protein family, though its function is not yet understood, consisted of a predicted signal sequence, single transmembrane region, and cytoplasmic kinase domain. RLKs are involved in hormonal response pathways, cell differentiation, plant growth and development, self-incompatibility, and symbiont and pathogen recognition. In this study, expression levels of RLG1, RLG5, RLG6, RLG#6, RLG8, RLG10, RLG17, RLG18 and RLG20 were analyzed by Real-time PCR, when rice (Oryzae sativa) was treated abiotic stress. The expression levels of all RLGs were compared each other by analyzed value of threshold cycles ($C_T$). Consequently, RLGs were suppressed by NaCl as salinity stress, and expression of each RLK genes were showed difference treated salicylic acid and wound, respectively. However, All RLGs were induced under low temperature condition. Therefore, our results indicate protection-function of RLK genes to be an early response of rice against cold weather.

• BMB Reports
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• v.39 no.1
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• pp.9-15
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• 2006

To prove whether error catastrophe /lethal mutagenesis is the primary antiviral mechanism of action of ribavirin against foot-and-mouth disease virus (FMDV). Ribavirin passage experiments were performed and supernatants of $Rp_1$ to $Rp_5$ were harvested. Morphological alterations as well as the levels of viral RNAs, proteins, and infectious particles in the BHK-21 cells infected using the supernatants of $Rp_1$ to $Rp_5$ and control were measured by microscope, real-time RT-PCR, western-blotting and plaque assays, respectively. The mutation frequency was measured by sequencing the complete P1- and 3D-encoding region of FMDV after a single round of virus infection from ribavirin-treated or untreated FMDV-infected cells. Ribavirin treatment for FMDV caused dramatically inhibition of multiplication in cell cultures. The levels of viral RNAs, proteins, and infectious particles in the BHK-21 cells infected were more greatly reduced along with the passage from $Rp_1$ to $Rp_5$, moreover, nucleocapsid protein could not be detected and no recovery of infectious virus in the supernatant or detection of intracellular viral RNA was observed at the $Rp_5$-infected cells. A high mutation rate, giving rise to an 8-and 11-fold increase in mutagenesis and resulting in some amino acid substitutions, was found in viral RNA synthesized at a single round of virus infection in the presence of ribavirin of $1000\;{\mu}M$ and caused a 99.7% loss in viral infectivity in contrast with parallel untreated control virus. These results suggest that the antiviral molecular mechanism of ribavirin is based on the lethal mutagenesis/error catastrophe, that is, the ribavirin is not merely an antiviral reagent but also an effective mutagen.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.373-390
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• 1998

35 theses conducted in Korea on the topic of senescence or anti-oxidant were classified on the basis of research methods, animals used for the experiments, and research items. Evaluating these research works with respect to the Free Radical Theory, the following conclusions were reached. 1. Of the 17 theses written in the Oriental medicine aspect, three theses used a single herb, nine theses used a complex prescription, four theses concentrated on the usage of the medicinal acupuncture, and one research paper focused on using scientific components. Common objection of these papers were on the verification of the efficacy of herbs. 2. Of the 18 these written in the Western medicine aspect, five theses used a single drug, seven theses conducted a research on the changes due to senescence, and 6 papers were on variety of topics. The main focus of these works were on the mechanism and pathology related to the senescence rather than on the suppression of senescence. 3. Among the theses written in the Oriental medicine perspective, a total of 48 herbs were utilized. 26 of these herbs has a tonification function on the Kidney. Six out of nine complex prescription mentioned above has a function of tonifying the Kidney. 4. With respect to the research subjects used on the experiments, 8 theses have used Senescence-Accelerated mice, 13 theses have used Sprague-Dawley mice, and remaining 7 papers have used human or other animals. 5. These are the categorization of the research items used: the weight (11) and weight changes of the visceral organs (9), the measurement of the content of peroxide-disease (9), the measurement of enzyme vitality (21), the blood and urine test (10), the experiment concerning immune system (3), the influence on the hepatic capability of metabolizing foreign substance (3), the effect on hepatic cell protection (3), the measurement of both the suppression of Free Radical and ability to create Free Radical (2), the measurement of effect of suppresing MDA(malondialdehyde) (4), the effect of eliminating DPPH Radical (2), and experiements about the functions (2). 6. The rate of vitalization of well known anti-oxidants such as Superoxide dismutase (SOD), Protein-bound SH, Nonprotein-bound SH, Glutathione(GSH), Catalase, and etc. were tested in 17 theses. Considering the conclusions mentioned above, the theses related to the senescence published in Korea elected different animals used for experiments, research items and the methods of research, the end result seems to be a lack of objectivity. Thus, I would argue that research methods to overcome such a deficiency need to be developed systematically.

• Journal of Life Science
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• v.12 no.6
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• pp.688-693
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• 2002

Molecular chaperones prevent the misfolding of newly synthesized polypeptides in the cell. The coexpression of molecular chaperones could be expected to improve the production of soluble and active recombinant proteins. In this study, the effect of coexpression of E. coli GroEL/ES chaperone on the active production of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in E. coli was investigated. Two plasmids, pTCGT1 and pGro7 in which the cgt and the groEL/ES genes are under the control of 77 promoter and araB promoter, respectively, were co-transformed into E. coli. With a series of cultures of recombinant E. coli cells, the optimal concentrations of IPTG and L-arabinose were found be 1 mM and 0.3 mg/$m\ell$, respectively. When IPTG and L-arabinose were added at 0.8~1.0 $OD_{600}$ and 0.4~0.5 $OD_{600}$, active CGTase production was increased significantly. This coexpression condition resulted in 1.5-fold increased level of soluble CGTase (0.7~0.73 unit/$m\ell$), compared to the level of CGTase in the single expression (0.36~0.56 unit/$m\ell$). An SDS-PACE analysis revealed that about 33.6% of CGTase in the total CGTase protein was found in the soluble fraction by coexpression of GroEL/ES chaperone.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.sup3
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• pp.179-183
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• 2016

Breast cancer is the most prevalent type of cancer among women around the world, and mortality is primarily caused by micro-metastatic disease. The complex mechanisms of breast cancer invasion and metastasis are intrinsically related to the malignant cell type so that early detection of micro-metastases can help prolongation of survival for patient. The aim of the present research work was evaluation of the expression status of mammoglobin protein as a candidate molecular marker in the negative sentinel lymph node (SLN). Fifty tumor specimens, and 50 normal adjacent breast tissue samples from the same patients were selected on the basis of having more than 10% tumor content for RNA extraction from SLNs. Tumor samples and normal adjacent breast tissue were archived in the form of frozen fresh tissue in liquid nitrogen. Real-time PCR was performed on a Bioner life express gradient thermal cycler system. Mammoglobin gene overexpression in breast cancer metastasis was investigated. Single marker results were mammaglobin 66.7% and CK19 50.0%, with 58.3% for the two in combination. Due to improved outcome with at least 3 genes (83.3%), it seems, triple marker evaluation will be most likely useful for detecting micro-metastases instead of studying separate genes.

• Korean Journal of Head & Neck Oncology
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• v.19 no.2
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• pp.121-126
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• 2003

Objectives: The c-kit gene encodes a transmembrane receptor-type tyrosine kinase, which is known to have a significant role in the normal migration and development of germ cells and melanocytes. In the previous studies of c-kit gene, c-kit expressions showed only in adenoid cystic carcinomas, lymphoepithelioma-like carcinomas and myoepithelial carcinomas, but not in others and mutation was not found in any types of salivary carcinoma. We investigate the c-kit expression which may be useful to differentiating adenoid cystic carcinomas from others, and mutation of the gene which may not be exist nor the mechanism of c-kit activation in salivary carcinomas. Material and Methods: The archival tissue samples from 42 salivary carcinomas of major and minor salivary glands were studied for c-kit expression by immunohistochemistry and gene mutation by polymerase chain reaction amplification and single strand conformational polymorphism. Results: The c-kit expressions were noted in 22/24 adenoid cystic carcinomas, 7/9 mucoepidermoid carcinomas, 2/3 acinic cell carcinomas, 3/4 malignant mixed tumors, and one undifferentiated carcinoma. The mutation of c-kit gene was found in 3/24 adenoid cystic carcinomas, 3/8 mucoepidermoid carcinomas, one acinic cell carcinoma, and 2/4 malignant mixed tumors. Conclusion: c-kit protein overexpression is seen in a variety of salivary gland carcinomas, and the mutation of the gene may be the mechanism of c-kit activation in these neoplasms.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.311-318
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• 2002

A new bacteriocin producing lactic acid bacteria having antagonistic activity against Lactobacillus plantarum, was isolated from Kimchi. It was identified as Leuconostoc mesenteroides, and designated as Leuconostoc mesenteroides B7. The bacteriocin from Leuconostoc mesenteroides B7 named as bacteriocin B7 was stable in the pH range $2.5{\sim}9.5$. Bacteriocin B7 was active over a wide temperature range from $4^{\circ}C$ to $120^{\circ}C$. It was inactivated by proteinase K, trypsin, ${\alpha}-chymotrypsin$, and protease treatments indicating its proteinous nature. Tricine-SDS-PAGE of the purified bacteriocin B7 showed the presence of a single band, having a molecular mass of about 3,500 dalton. Mixed culture of the producer and the indicator, Lb. plantarum KFRI 464 or Lb. delbruekii KFRI 347, increased production of bacteriocin B7. This result suggested the presence of bacteriocin inducing factor in the indicator strain. The inducing factor was localized in cell debris and intracellular faction of the indicator cell, Lb. plantarum KFRI 464. Treatment of the inducing factor with proteinase K destroyed inducing activity. This result strongly suggested that the inducing factor is a protein.

• Biomolecules & Therapeutics
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• v.28 no.5
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• pp.443-455
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• 2020

The thioredoxin (Trx) system plays critical roles in regulating intracellular redox levels and defending organisms against oxidative stress. Recent studies indicated that Trx reductase (TrxR) was overexpressed in various types of human cancer cells indicating that the Trx-TrxR system may be a potential target for anti-cancer drug development. This study investigated the synergistic effect of auranofin, a TrxR-specific inhibitor, on sulforaphane-mediated apoptotic cell death using Hep3B cells. The results showed that sulforaphane significantly enhanced auranofin-induced apoptosis by inhibiting TrxR activity and cell proliferation compared to either single treatment. The synergistic effect of sulforaphane and auranofin on apoptosis was evidenced by an increased annexin-V-positive cells and Sub-G1 cells. The induction of apoptosis by the combined treatment caused the loss of mitochondrial membrane potential (ΔΨm) and upregulation of Bax. In addition, the proteolytic activities of caspases (-3, -8, and -9) and the degradation of poly (ADP-ribose) polymerase, a substrate protein of activated caspase-3, were also higher in the combined treatment. Moreover, combined treatment induced excessive generation of reactive oxygen species (ROS). However, treatment with N-acetyl-L-cysteine, a ROS scavenger, reduced combined treatment-induced ROS production and apoptosis. Thereby, these results deduce that ROS played a pivotal role in apoptosis induced by auranofin and sulforaphane. Furthermore, apoptosis induced by auranofin and sulforaphane was significantly increased through inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway. Taken together, the present study demonstrated that down-regulation of TrxR activity contributed to the synergistic effect of auranofin and sulforaphane on apoptosis through ROS production and inhibition of PI3K/Akt signaling pathway.

• Journal of Life Science
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• v.33 no.2
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• pp.191-198
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• 2023

As a member of the focal adhesion complex of the plasma membrane, Src is a nonreceptor tyrosine kinase that controls cell adhesion and motility. However, how Src activity is regulated in the plasma membrane microdomain in response to components of the extracellular matrix (ECM) remains unclear. This study compared and investigated the activity of Src in response to three representative ECM proteins: collagen type 1, fibronectin, and laminin. Genetically encoded FRET-based Src biosensors for plasma membrane subdomains were used. FRET-based biosensors allow the real-time analysis of protein activity in living cells based on their high spatiotemporal resolution. The results showed that Src activity was maintained at a high level under all ECM conditions of the lipid raft, and there was no significant difference between the ECM conditions. In contrast, Src activity was maintained at a low level in the non-lipid raft membrane. In addition, the Src activity of lipid rafts remained significantly higher than that of non-lipid raft regions under the same ECM conditions. In conclusion, this study demonstrates that Src activity can be controlled differently by lipid rafts and non-lipid raft microdomains.