• Korean Journal of Head & Neck Oncology
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• v.33 no.1
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• pp.21-29
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• 2017

Methylation of CpG islands in the promoter region of genes acts as a significant mechanism of epigenetic gene silencing in head and neck squamous cell carcinoma (HNSCC). DNA methylation markers are particularly advantageous because DNA methylation is an early event in tumorigenesis, and the epigenetic modification, 5-methylcytosine, is a stable mark. In the present study, we assessed the genome-wide preliminary screening and were to identify novel methylation biomarker candidate in HNSCC. Genome-wide methylation analysis was performed on 10 HNSCC tumors using the Methylated DNA Isolation Assay (MeDIA) CpG island microarray. Validation was done using immunohistochemistry using tissue microarray of 135 independent HNSCC tumors. In addition, in vitro proliferation, migration/invasion assays, RT-PCR and immunoblotting were performed to elucidate molecular regulating mechanisms. Our preliminary validation using CpG microarray data set, immunohisto-chemistry for HNSCC tumor tissues and in vitro functional assays revealed that methylation of the Homeobox B5 (HOXB5) and H6 Family Homeobox 2 (HMX2) could be possible novel methylation biomarkers in HNSCC.

• KSBB Journal
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• v.20 no.3
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• pp.220-227
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• 2005

Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. To understand the potential target biosynthetic genes critical for the doxorubicin everproduction, a doxorubicin-specific DNA microarray chip was fabricated and applied to reveal the growth-phase-dependent expression profiles of biosynthetic genes from two doxorubicin-overproducing strains along with the wild-type strain. Two doxorubicin-overproducing 5. peucetius strains were generated via over-expression of a dnrl (a doxorubicin-specific positive regulatory gene) and a doxA (a gene involved in the conversion from daunorubicin to doxorubicin) using a streptomycetes high expression vector containing a strong ermE promoter. Each doxorubicin-overproducing strain was quantitatively compared with the wild-type doxorubicin producer based on the growth-phase-dependent doxorubicin productivity as well as doxorubicin biosynthetic gene expression profiles. The doxorubicin-specific DNA microarray chip data revealed the early-and-steady expressions of the doxorubicin-specific regulatory gene (dnrl), the doxorubicin resistance genes (drrA, drrB, drrC), and the doxorubicin deoxysugar biosynthetic gene (dnmL) are critical for the doxorubicin overproduction in S. peucetius. These results provide that the relationship between the growth-phase-dependent doxorubicin productivity and the doxorubicin biosynthetic gene expression profiles should lead us a rational design of molecular genetic strain improvement strategy.

• Genomics & Informatics
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• v.12 no.4
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• pp.240-246
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• 2014

Mutation in HNF1B, the hepatocyte nuclear factor-$1{\beta}$ (HNF-$1{\beta}$) gene, results in maturity-onset diabetes of the young (MODY) 5, which is characterized by gradual impairment of insulin secretion. However, the functional role of HNF-$1{\beta}$ in insulin secretion and glucose metabolism is not fully understood. We identified a family with early-onset diabetes that fulfilled the criteria of MODY. Sanger sequencing revealed that a heterozygous P159L (CCT to CTT in codon 159 in the DNA-binding domain) mutation in HNF1B was segregated according to the affected status. To investigate the functional consequences of this HNF1B mutation, we generated a P159L HNF1B construct. The wild-type and mutant HNF1B constructs were transfected into COS-7 cells in the presence of the promoter sequence of human glucose transporter type 2 (GLUT2). The luciferase reporter assay revealed that P159L HNF1B had decreased transcriptional activity compared to wild-type (p < 0.05). Electrophoretic mobility shift assay showed reduced DNA binding activity of P159L HNF1B. In the MIN6 pancreatic ${\beta}$-cell line, overexpression of the P159L mutant was significantly associated with decreased mRNA levels of GLUT2 compared to wild-type (p < 0.05). However, INS expression was not different between the wild-type and mutant HNF1B constructs. These findings suggests that the impaired insulin secretion in this family with the P159L HNF1B mutation may be related to altered GLUT2 expression in ${\beta}$-cells rather than decreased insulin gene expression. In conclusion, we have identified a Korean family with an HNF1B mutation and characterized its effect on the pathogenesis of diabetes.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.596-606
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• 2019

N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ${\Delta}swrI$ with or without supplementing exogenous N-hexanoyl-L-homoserine lactone ($C_6-HSL$) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing $C_6-HSL$. Furthermore, fermentation product analysis indicated that ${\Delta}swrI$ could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ${\Delta}swrI$ appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, ${\alpha}$-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.

• Fisheries and Aquatic Sciences
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• v.22 no.9
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• pp.20.1-20.8
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• 2019

Background: Despite the favorable geo-climatic potential of Cameroon, the national production of tilapia remains low due to poor tilapia growth reported by fish farmers. One of the underlying reasons is the early female maturation at a very small size and precocious breeding in earthen ponds, resulting in overpopulation which leads to stunted growth and therefore to the production of unmarketable fish size. Studies have shown that dietary supplementation of G. kola enhanced growth in young Clarias gariepinus and Oreochromis niloticus. It was also reported that G. kola inhibited spawning in Tilapia adult females. Therefore, this study sought to assess the effects of Garcinia kola as growth promoter and inhibitor of gonadal development in young Oreochromis niloticus. Methods: A total of 108 juveniles weighing $13.32{\pm}0.62g$ were randomly distributed in 9 hapas of 12 fishes each (9 females and 3 males) and fed for 70 days with three isonitrogenous diets, 40% crude protein with increasing Garcinia kola supplementation levels of 0 (normal diet), 6% and 10% (experimental diets). Physico-chemical parameters of the water (temperature, dissolved oxygen, pH, nitrate, nitrite, ammonia, and transparency) were measured twice a week. Every 14 days, fish were harvested, counted, and weighed. At the end of the experiment, three fish of each sex per replicate were sacrificed and their gonad and liver collected and weighed. Data were statistically analyzed using one-way analysis of variance repeated measure followed by Newman-Keuls multiple tests. Results: The results showed that all physico-chemical parameters of the water were within the recommended values for Tilapia culture. Tilapia fed 6% Garcinia kola supplemented diet displayed higher final body weight in males ($38.60{\pm}3.50g$) and females ($36.77{\pm}3.62g$) compared to those receiving normal diet ($36.23{\pm}1.36g$ and $25.87{\pm}3.32g$; respectively to the final body weight in males and females). The gonadosomatic index and hepatosomatic index indicated no significant variation in males while in females, these were significantly low in the experimental fish compared to control fish. Conclusion: The results of this study demonstrated that supplementation of G. kola seeds in diets of young Tilapia improved growth performance and impaired gonadal development in females.

• BMB Reports
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• v.56 no.10
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• pp.563-568
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• 2023

DNA methylation regulates gene expression and contributes to tumorigenesis in the early stages of cancer. In colorectal cancer (CRC), CpG island methylator phenotype (CIMP) is recognized as a distinct subset that is associated with specific molecular and clinical features. In this study, we investigated the genome-wide DNA methylation patterns among patients with CRC. The methylation data of 1 unmatched normal, 142 adjacent normal, and 294 tumor samples were analyzed. We identified 40,003 differentially methylated positions with 6,933 (79.8%) hypermethylated and 16,145 (51.6%) hypomethylated probes in the genic region. Hypermethylated probes were predominantly found in promoter-like regions, CpG islands, and N shore sites; hypomethylated probes were enriched in open-sea regions. CRC tumors were categorized into three CIMP subgroups, with 90 (30.6%) in the CIMP-high (CIMP-H), 115 (39.1%) in the CIMP-low (CIMP-L), and 89 (30.3%) in the non-CIMP group. The CIMP-H group was associated with microsatellite instability-high tumors, hypermethylation of MLH1, older age, and right-sided tumors. Our results showed that genome-wide methylation analyses classified patients with CRC into three subgroups according to CIMP levels, with clinical and molecular features consistent with previous data.

• Journal of Sericultural and Entomological Science
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• no.11
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• pp.31-44
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• 1970

This experiment is conducted fer method cf bring to perfectly from defective cutting sapling 25 a production method of sapling which is to substitute for conventional graf ting. 1. In green wood cutting root comes out after 15 days of cutting with nearly straight development of root system; after 40 days of cutting, roots with total length of 1119 mms 43 roots, and 5.4 grs a root in total weight obtained. 2. Survival percentage of green wood cutting ranges between 56％ and 90％, average 73％ of that and it varies with natural characteristics of varietes. The results show variety of Gaeryang-Suban and Iljiroe with 80％ as a best ones in contrast with the variety of Shipyung. 3. The varietics or Gaeryang-Suban. Iljiroe, Suwon-Sang No.4, Rosang makes of much more roots than Yongchonchuwoo and Shipyung do. 4. Root ability made good number of roots commercially, when cutting is conducted soaking in 0.01 ％ NAA solution or 0.02％ NAA solution for 2 or 3 seconds as a chemical promoter. 5. Economical measures for increase of scion adapted 1) 2 scions from a green wood 50 cm long should be taken and basal pan of scion at its middle portion should be cut right under the node. 2) Scions below 50cm long may be used. Small scions or growthceased shoots h3ve also considerable root ability enough to be used. 3) Thus far, up 100,000 scions might be produced in 10 a. 6. We can find number of root increased, when cutting the opposite side in obliquity manner at 450. 7. When 110,000 saplings in 10 a. for the production of bring to perfectly from defective cutting sapling planted, quality of stocks does not go to bad condition without any obstacles in practical use. 8. Although the times or grafting delayed until middle of July, quality of sapling goes just a little down. Grafting may be conducted from early June to middle of July separately in several times, and the green wood of prunned mulberry in spring is available for a scion after end of June. 9. 10 grs weight of defective cutting sapling makes 95％ of complete sapling, otherwise 5 or 10 grs in weight of one becomes 80％ of complete sapling with its quality as similar as grafted one. 10. When the sapling planted, its branches should be cut, leaving 3 or 4 buds at the bottom of new branches. 11. In view of economical stand point, production cost of bring to perfectly from defective cutting sapling obtains 52％ of grafting cost.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.140-145
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• 2012

The flowering locus T (FT) gene, of which expression will be controlled at high temperature by heat shock promoter (it printed as to HSproFT), was introduced into spray-type chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) 2 cultivars ('Pink PangPang' and 'Pink Pride' by co-cultivation with Agrobacterium tumefaciens strain C58C1 harboring pCAMBIA2300 containing the HSproFT gene. After leaf segments of the 2 cultivars were infected with the A. tumafaciens with C58C1 as explants, shoots were regenerated from the explants cultured on the $1^{st}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 10 mg/L kanamycin + 0.7% plant agar, pH 5.8). The shoots were transferred into the $2^{nd}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 20 mg/L kanamycin + 0.7% plant agar, pH 5.8). One hundred seventeen plantlets from 'Pink PangPang' and 5 ones from 'Pink Pride' were confirmed as transformants by PCR analysis. Twenty six of the transformants and non-transformants were acclimatized and established well in a green house. Eights of 26 transgenic lines showed flower bud 1.7~10 days earlier than nontransgenic plants, and 24 of them flowered 1~6 days earlier than non- transgenic plants. The shape and color of flower of all HSproFT-transgenic lines were not different with those of non- transgenic plants.