Aberrant expression of microRNAs (miRNAs) plays important roles in carcinogenesis and tumor progression. However, the expression and biological role of miR-301b in triple-negative breast cancer (TNBC) remains unclear. Here we aimed to evaluate the roles and mechanisms of miR-301b in TNBC cells. miR-301b expression was assessed in TNBC specimens and cell lines by quantitative Real-Time PCR (qRT-PCR). TNBC cells were transfected with miR-301b mimics, inhibitors or Cylindromatosis (CYLD) small interfering RNA (siRNA) using Lipofectamine 2000. The functional roles of miR-301b were determined by cell proliferation, colony formation, and apoptosis assays. Western blots and qRT-PCR were used to measure the expression of mRNAs and proteins in the cells. We found that miR-301b was upregulated in TNBC specimens and cell lines. Overexpression of miR-301b promoted cell proliferation in TNBC cells, while inhibited the apoptosis induced by 5-FU. CYLD was downregulated by miR-301b at both mRNA and protein levels in TNBC cells. Dual-luciferase report assay confirmed that miR-301b downregulated CYLD by direct interaction with the 3'-untranslated region(3'-UTR) of CYLD mRNA. $NF-{\kappa}B$ activation was mechanistically associated with miR-301b-mediated downregulation of CYLD. However, inhibition of miR-301b reversed all the effects of miR-301b. In conclusion, miR-301b plays an oncogenic role in TNBC possibly by downregulating CYLD and subsequently activating $NF-{\kappa}B$ p65, and this may provide a novel therapeutic approach for TNBC.
Proceedings of the Korean Society for Bioinformatics Conference
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2005.09a
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pp.267-271
/
2005
The goal of data mining is to extract new and useful knowledge from large scale datasets. As the amount of available data grows explosively, it became vitally important to develop faster data mining algorithms for various types of data. Recently, an interest in developing data mining algorithms that operate on graphs has been increased. Especially, mining frequent patterns from structured data such as graphs has been concerned by many research groups. A graph is a highly adaptable representation scheme that used in many domains including chemistry, bioinformatics and physics. For example, the chemical structure of a given substance can be modelled by an undirected labelled graph in which each node corresponds to an atom and each edge corresponds to a chemical bond between atoms. Internet can also be modelled as a directed graph in which each node corresponds to an web site and each edge corresponds to a hypertext link between web sites. Notably in bioinformatics area, various kinds of newly discovered data such as gene regulation networks or protein interaction networks could be modelled as graphs. There have been a number of attempts to find useful knowledge from these graph structured data. One of the most powerful analysis tool for graph structured data is frequent subgraph analysis. Recurring patterns in graph data can provide incomparable insights into that graph data. However, to find recurring subgraphs is extremely expensive in computational side. At the core of the problem, there are two computationally challenging problems. 1) Subgraph isomorphism and 2) Enumeration of subgraphs. Problems related to the former are subgraph isomorphism problem (Is graph A contains graph B?) and graph isomorphism problem(Are two graphs A and B the same or not?). Even these simplified versions of the subgraph mining problem are known to be NP-complete or Polymorphism-complete and no polynomial time algorithm has been existed so far. The later is also a difficult problem. We should generate all of 2$^n$ subgraphs if there is no constraint where n is the number of vertices of the input graph. In order to find frequent subgraphs from larger graph database, it is essential to give appropriate constraint to the subgraphs to find. Most of the current approaches are focus on the frequencies of a subgraph: the higher the frequency of a graph is, the more attentions should be given to that graph. Recently, several algorithms which use level by level approaches to find frequent subgraphs have been developed. Some of the recently emerging applications suggest that other constraints such as connectivity also could be useful in mining subgraphs : more strongly connected parts of a graph are more informative. If we restrict the set of subgraphs to mine to more strongly connected parts, its computational complexity could be decreased significantly. In this paper, we present an efficient algorithm to mine frequent subgraphs that are more strongly connected. Experimental study shows that the algorithm is scaling to larger graphs which have more than ten thousand vertices.
Kim Jong Kwan;Kim Hyoung Soon;Bae Young Chun;Lee Sang Min;Kim Kyung Yo;Joo Jong Cheon
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.4
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pp.1192-1198
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2004
Sasang Constitutional Medicine is a major branch of Korean Traditional Medicine. The differences of disease susceptibility to be shown in Sasang constitution may be due to genetic factors. Therefore, I examined interrelationship among cerebral infarction (CI), apolipoprotein E (apo E) gene polymorphism, and Sasang constitutional classification. Apo E is a key protein modulating the highly atherogenic apoB containing lipoproteins and is a candidate gene for the development of coronary artery disease (CAD). The ε2 and/or ε4 alleles were the first to be implicated in premature CAD, which resulted in this polymorphism being extensively studied. I investigated the association between apo E genotype and CI by case-control study in a Korean population. I also classified CI patients and control group into groups according to Sasang Constitutional Medicine. 218 CI patients and 379 controls without CI were examined. Apo E genotype was determined by 8% polyacrylamide gel separation after DNA amplification. A frequency of apo E ε3/ε3 in the apo E genotype distribution was higher in the CI patients compared with that in controls. Also, it was widely known that Taeumin was easily attacked with CI, but there was no association between apo E polymorphim and Taeumin. However, the Taeumin constitution did not enhance the relative risk for CI in the subjects with apo E ε2 and/or ε4 alleles. No differences in the apo E genotypes frequencies were observed in the Taeumin compared with that in the other constitutions. In addition, I investigated whether the DD(deletion/deletion) or ID(insertion/deletion) genotype of angiotensin converting enzyme (ACE) gene, a candidate gene for CI, was associated with CI, Taeumin constitution, and apo E polymorphism. As a result, the frequency of Taeumin constitution was significantly higher in CI patients with both apo E ε3/ε4 and ACE ID/DD genotypes than in the remaining Sasang constitutions. In summary, it was concluded that the apo E polymorphism is a major risk factor for CI in Koreans and the ACE ID/DD genotype enhanced the relative risk for CI in the subjects with apo E ε3/ε4 genotype and Taeumin constitution.
The effect of metolachlor [2-chloro-N-(2-ethyl 1-6-methylphenyl)-N-(2-methoxy-l-methylethyl)-acetamide] on starch, sugar and protein content in relation with ${\alpha}$-amylase activity, and its interaction with GA in the germinating stage of rice were determined. The distinctive phytotoxic symptom was the failure of the primary leaf of rice to break the coleoptile. An inhibitory effect of metolachlor was nullified by the external application of GA $10^{-3}$M. A significantly slower starch degradation accompanying the lower content of sugars was observed in the gain of the metolachlor treated seedlings. Further, the total ${\alpha}$-amylase activity was significantly lower in the grain of rice seedlings treated with metolachlor than that of the untreated one, and the lower total ${\alpha}$-amylase activity could be due to an inhibition of ${\alpha}$-amylase formation.
The selection of multienzyme complex-producing bacteria under aerobic condition was conducted for improving the degradation of lignocellulosic substances. The criteria for selection were cellulase and xylanase enzyme production, the presence of cellulose-binding domains and/or xylan-binding domains in enzymes to bind to insoluble substances, the adhesion of bacterial cells to insoluble substances, and the production of multiple cellulases and xylanases in a form of a high molecular weight complex. Among the six Bacillus strains, isolated from various sources and deposited in our laboratory, Paenibacillus curdlanolyticus B-6 strain was the best producer of cellulase and xylanase enzymes, which have both cellulose-binding factors (CBFs) and xylan-binding factors (XBFs). Moreover, multiple carboxymethyl cellulases (CMCases) and xylanases were produced by the strain B-6. The zymograms analysis showed at least 9 types of xylanases and 6 types of CMCases associated in a protein band of xylanase and cellulase with high molecular weight. These cells also enabled to adhere to both avicel and insoluble xylan, which were analyzed by scanning electron microscopy. The results indicated that the strain B-6 produced the multienzyme complex, which may be cellulosome or xylanosome. Thus, P. curdlanolyticus B-6 was selected to study the role and interaction between the enzymes and their substrates and the cooperation of multiple enzymes to enhance the hydrolysis due to the complex structure for efficient cellulases and xylanases degradation of insoluble polysaccharides.
The interactive effect of dietary fat supplementation and milk yield level on dairy cows performance under heat stress has not been thoroughly investigated. The purpose of this study was to evaluate the effect of production level, the source of fat supplements and their interaction on dairy cows performance under heat stress. In this study, 64 Holstein multiparous cows were divided into 2 groups and received one of two rations having either calcium salts of fatty acids (Ca-FA) or high-palmitic acid (PA) supplements (2.8% of DM; dry matter). After completing the experiment and based on maturity-equivalent milk, cows were divided into two groups of high-yielding (14,633 kg) and medium-yielding (11,616 kg). Average temperature humidity index (THI) was 71 during the trial period. Apparent digestibility of dry matter (p = 0.04), organic matter (p = 0.05), and neutral detergent fiber (NDF; p = 0.04) for cows fed Ca-FA were greater than cows fed PA. The milk fat content in high-producing cows was 0.3% greater than medium-producing cows (p = 0.03). The milk protein content in cows fed Ca-FA was greater than cows fed PA (p < 0.01). High-producing cows had greater serum cholesterol (p = 0.02) than medium-producing cows. The cows fed PA tended to have a greater BUN than cows fed Ca-FA (p = 0.06). Alanine aminotransferase and aspartate aminotransferase tended to be increased by PA, which indicates that cows in PA treatment may have experienced more adverse effect on the liver function than cows on Ca-FA. Therefore, under heat stress and in 90 d trial, milk production level does not affect the cows' response to PA or Ca-FA. Although cows fed Ca-FA received lower energy than those fed PA, they compensated for this shortage likely with increasing the digestibility and produced a similar amount of milk.
The consumption of foods rich in TDF should not be associated with impaired mineral absorp-tion and long-term mineral status. In surveys of populations consuming high amounts of TDF e.g Third World populations and vegetarinas gross deficiencies in mineral nutrition have not been noted. If mineral status is low among these groups it is most likely caused by the inadequacy or imbalance of the diet and not by the TDF. The key word is interaction which should be inte-rpreted in dietary imbalances that produce nut-rient deficiencies. There are no strong data to support the concept that TDF inhibits mineral absorption through a binding chelation mechanism. Limited data sug-gest that positively charged groups on polymers such as chitosan and cholestyramine will decrease iron absorption in humans and animals. Because TDF does not contain positively charged groups future research should be directed at the possible role of protein consumed along with TDF and the combination of effects on mineral nutrition Phytic acid is acknowledged as a potent chela-tor of zinc. However its association with zinc and its propensity to lower Zn bioavaiability may enhance the absorption of other elements notably copper and iron. The importance of interactions among nutrients including TDF will gain addi-tional attention in the scientific community. Soluble and insoluble dietary fiber function di-fferently in the intestine. Insoluble fibers accele-rate movement through the intestine. Soluble die-tary fibers appear to regulated blood concentra-tions of glucose and cholesterol albeit by some unknown mechanism. In creased viscosity produ-ced by the SDF in the intestine may provide an explanation of how this class of polymers affects plasma glucose cholesterol and other nutrients. Employing a double-perfusion technique in the rat we demonstrated that viscosity produced by SDF will delay transfer of zinc into the circulatory system. This delayed absorption should not be interpreted as decreased utilization. A great deal of additional research is required to prove the importance of luminaly viscosity produced by SDF on slowing nutrient absorption or regulating bllod nutrient homeostasis. Increased intake of TDF in the total human diet appears desirable. A dietary intake of 35g/day should not be considered to have a negative effect on mineral absorption. It is important to educate people that an intake of more than 35g TDF/day may cause an imbalance in the diet that can adve-rsely affect mineral utilization. Acknowledgments. Appreciation is given to Dr. George V. Vahouny(deceased) who was intense a great competitor in and out of science and who gave the author inspiration Portions of this work were supported by the University of Missouri Ag-ricultural Station and by a grant from the Univer-rch Support Grant RR 07053 from the National Institutes of Health. Contribution of the Missouri Agriculatural Experiments Station Journal Series No. 10747.
A study was conducted to determine the effects of restricted feeding and feed form on the growth performance, characteristics of carcass and digestive tract, and days to first egg of Japanese quail (JQ, Coturnix coturnix japonica). A total of 240 oneweek-old JQ chicks were allocated randomly into 4 experimental groups that consisted of 3 replicates according to a 2${\times}$2 factorial arrangement for two feeding methods (ad libitum, AF and restricted feeding, RF) and two diet forms (mash, MD and crumble, CD). The JQ chicks were placed in a room with floor battery brooders and fed a commercial starter diet from 7 to 14 d of age. According to the experimental design, four treatments (1: ad libitum MD, 2: restricted MD, 3: ad libitum CD, and 4: restricted CD) were applied. Feed restriction was applied by 30% reduction of ad libitum feed intake for both MD and CD from 15 to 28 d of age. All birds were fed ad libitum with treatment diets from 29 d of age until the first laid egg seen (45 d of age). The commercial starter diet, MD and CD were in the same nutrient content (240 g crude protein with 13.4 MJ ME per kg diet). The body weight and overall feed conversion ratio (g feed/g gain) were higher (p<0.05) for the AF quails than the RF at 42 d of age. Carcass weights, dressing percentage and percentage yields of breast and back were similar for AF and RF groups at 42 d of age. The RF delayed (p<0.05) onset of egg production 2 days compared to the AF. Quail fed with the CD showed higher value (p<0.05) for carcass weight and dressing percentage at 42 d of age compared to birds fed with the MD. The interaction effect of feeding method${\times}$feed form on any of the studied parameters was not significant. The results suggest that feed restriction as in the present study can achieve a better feed conversion without reduction in carcass weight, and a significant benefit of feeding the crumble diet over the mash diet was obtained in terms of carcass weight in the JQ.
Kim, Kee-Soon;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.26
no.4
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pp.355-362
/
2000
Neoplastic growth is characterized by alterations of oncogenes and antioncogenes. The interaction between activated oncogenes and functional deletion of antioncogene appears to be the driving force directing normal cells to uncontrolled growth resulting in tumor. In addition to those genes mentioned, other genes controlling the entry of cells into the cell cycle have recently been implicated in cancer development. The overexpression of the cyclin D1 gene, which has been mapped to 11q13, either by gene rearrangement or amplification has been noted in various malignant tumors. The product of the cyclin D1 gene forms a complex with cyclin-dependent protein kinases(CDK4) that governs a key transition in the cell cycle. The relationships between the overexpression of cyclin D1 assessed by immunihistochemistry and the amplification of the cyclin D1 gene by differential polymerase chain reaction(DPCR) using primers for dopamin D2 receptor gene in 13 cases of squamous cell carcinomas of the oral cavity have been studied. The semiquantitative assay of cyclin D1 amplification has been made by cyclin D1/dopamin D2 receptor(CD/DR) ratio. The results were as follows; 1. In the normal tissue and the tumor, the CD/DR ratios were 0.82 and 1.36 respectively. This implicates 1.65-fold amplification of cyclin D1 gene in tumor compared to that in normal tissue. 2. The tumor tissue which showed overexpression of cyclin D1 by immunohistochemistry revealed 2-fold amplification of cyclin D1 compared to the normal tissue. 3. The tumor tissue which showed mild expression of cyclin D1 by immunihistochemistry revealed 1.7-fold amplification of cyclin D compared to the normal tissue. 4. The cyclin D1 was overexpressed in the tumor tissue at the rate of 38%. Above results suggest that cyclin D1 has close correlation with the development of carcinoma in the oral cavity. But further studies were needed to elucidate the carcinogeneic mechanisms by comparative studies among cyclin D1, pRb and p53.
In the present report, a p[ossibility of the interaction fo Ser-33 and Asp-112 residues in folding of tryptophan synthase $\alpha$ subunit was explored by examining the effect of single or double substitution of these residues on folding of $\alpha$ subunit in E. coli. $\alpha$ subunit of which Ser-33 was substituted with Leu (SL33) was accumulated as insoluble aggregate form, when overproduced in E. coli, whereas $\alpha$ subunit of which Asp-112 was replaced by Asn (DN112) or Gly (DG112) was accumulated as soluble form to the similar extent as wild type $\alpha$ subunit was. When these alterations were combined into one protein, the synergistic effect of residues 33 and 112 on the amount of aggregate form was shown. The amount of doubly altered SL33/DG112 $\alpha$ subunit as aggregate form was increased 5-13 fold that of SL33 $\alpha$ subunit, and the amount of SL33/DG112 $\alpha$ subunit as aggregate form was decreased 3-4 fold that of SL33 $\alpha$ subunit. Aggregates are derived from the specific association of partially folded or unassembled subunits in the folding process. Therefore, this result suggests that residues 33 and 112 of $\alpha$ subunit may unteract during the folding of this enzyme in E. coli.
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