• Molecules and Cells
• /
• v.45 no.10
• /
• pp.718-728
• /
• 2022

Splicing factor B subunit 4 (SF3B4), a component of the U2-pre-mRNA spliceosomal complex, contributes to tumorigenesis in several types of tumors. However, the oncogenic potential of SF3B4 in lung cancer has not yet been determined. The in vivo expression profiles of SF3B4 in non-small cell lung cancer (NSCLC) from publicly available data revealed a significant increase in SF3B4 expression in tumor tissues compared to that in normal tissues. The impact of SF3B4 deletion on the growth of NSCLC cells was determined using a siRNA strategy in A549 lung adenocarcinoma cells. SF3B4 silencing resulted in marked retardation of the A549 cell proliferation, accompanied by the accumulation of cells at the G0/G1 phase and increased expression of p27, p21, and p53. Double knockdown of SF3B4 and p53 resulted in the restoration of p21 expression and partial recovery of cell proliferation, indicating that the p53/p21 axis is involved, at least in part, in the SF3B4-mediated regulation of A549 cell proliferation. We also provided ubiquitination factor E4B (UBE4B) is essential for p53 accumulation after SF3B4 depletion based on followings. First, co-immunoprecipitation showed that SF3B4 interacts with UBE4B. Furthermore, UBE4B levels were decreased by SF3B4 depletion. UBE4B depletion, in turn, reproduced the outcome of SF3B4 depletion, including reduction of polyubiquitinated p53 levels, subsequent induction of p53/p21 and p27, and proliferation retardation. Collectively, our findings indicate the important role of SF3B4 in the regulation of A549 cell proliferation through the UBE4B/p53/p21 axis and p27, implicating the therapeutic strategies for NSCLC targeting SF3B4 and UBE4B.

• Korean Journal of Microbiology
• /
• v.37 no.4
• /
• pp.253-258
• /
• 2001

A Bacillus circulans KCTC3004 $\beta$-1,3-glucanase gene contained in a recombinant plasmid pLM460 derived from subcloning the original recombinant plasmid pLM530 was trasferred into a new shuttle vector plasmid pLMS1180 by ligating linearized DNAs of pLM460 and pUB110. B. subtilis RM125 and B. megaterium ATCC14945 transformed with pLMS1180 produced the $\beta$-1,3-glucanase substantially. Most of the enzyme was produced during the exponential growth period. The maxium activities of the $\beta$-1,3-glucanase produced by the Bacillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125 (pLM1180) enzyme showed the activity 14 times higher than that of the gene donor cells, followed by the B. megaterium ATCC14945 (pLMS 1180) enzyme with activity 5 times higher than that of the gene donor cells. While E. coli secreted about 7% of the produced enzyme, B. subtilis excreted the enzyme into the medium wholly and B. megaterium about 97% of the total product. The SDS-PAGE of this enzyme produced in E. coli (pLMS1180), B subtilis (pLMS1180) or B. megaterium (pLMS1180) indicated a molecular weight of 38,000. The enzymes overproduced in three different host cells hydrolyzed laminarin to produce mainly laminaribiose, laminaritriose, and laminarioligosaccharides. The plasmid pLMS1180 was stable in B. megaterium, E. coli, but was unstable in B. subtilis.

• Korean Journal of Clinical Laboratory Science
• /
• v.52 no.1
• /
• pp.69-77
• /
• 2020

The tumor suppressor gene, p53, is inactivated in the human hepatocellular carcinoma cells line, Hep3B. Berberine has been reported to inhibit the proliferation of cancer cells. This study examined whether apoptosis was induced in berberine-treated Hep3B cells and observed the association between apoptosis and the expression of p53 and DNA methyltransferase (DNMT). The cell viability was measured using an MTT assay. Apoptosis of Hep3B was measured using annexin V flow cytometry. Berberine-treated cells were examined for their DNMT enzymatic activity, mRNA expression, and protein synthesis. The p53 levels were examined by Western blot analysis. The berberine treatment resulted in increased Hep3B cell death and apoptosis in a time- and dose-dependent manner. The DNMT3b activity, mRNA expression, and protein levels all decreased after the berberine treatment. In contrast, the p53 protein levels increased with a concomitant decrease in DNMT3b. No change in the expression of ERK was observed, but the P-ERK levels decreased in a dose dependent manner. These results indicate that a treatment of Hep3B cells with berberine can reduce the expression of DNMT3b, leading to an increase in the tumor suppressant gene p53 and an increase in cell apoptosis. This shows that berberine can effectively suppress the proliferation of liver cancer cells.

• The Korean Journal of Zoology
• /
• v.31 no.2
• /
• pp.142-146
• /
• 1988

Metabolism of benzo(a)pyrene, the most thoroughly studied PAH, was studied in mouse placental microsomes incubated with $^3$H-labeled B(a)P. B(a)P metabolites were separated using HPLC fitted with a C18- $\mu$ Bondapak column. The single major metabolite by mouse placental microsomes induced by B(a)P was 7, 8-diol B(a)P, while 4, 5-diol B(a)P, 3-OH and quinones constituted minor metabolites. Treatment with 3-methyl-cholanthrene to mice resulted in indudion of hydroxy B(a)P and quinone compounds. Phenobarbital treated mouse placental microsomes also showed elevated level of B(a)P metabolism with 7, 8-diol B(a)P as a major metabolite.

• The Journal of Advanced Prosthodontics
• /
• v.14 no.5
• /
• pp.315-323
• /
• 2022

PURPOSE. The purpose of this study was to investigate the effect of barium silicate filler contents on mechanical properties of resin nanoceramics (RNCs) for additive manufacturing (AM). MATERIALS AND METHODS. Additively manufactured RNC specimens were divided into 4 groups depending on the content of ceramic fillers and polymers: 0% barium silicate and 100% polymer (B0/P10, control group); 50% barium silicate and 50% polymer (B5/P5); 60% barium silicate and 40% polymer (B6/P4); 67% barium silicate and 33% polymer (B6.7/P3.3). The compressive strength (n = 15) and fracture toughness (n = 12) of the specimens were measured, and scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDS) analyses were performed. Independent sample Kruskal-Wallis tests were performed on the compressive strength and fracture toughness test results, and the significance of each group was analyzed at the 95% confidence interval through post-tests using the Bonferroni's method. RESULTS. B6/P4 and B6.7/P3.3 exhibited much higher yield strength than B0/P10 and B5/P5 (P < .05). Compared to the control group (B0/P10), the other three groups exhibited higher ultimate strength (P < .05). The fracture toughness of B6/P4 and B6.7/P3.3 were similar (P > .05). The content of barium silicate and fracture toughness showed a positive correlation coefficient (R = 0.582). SEM and EDS analyses revealed the presence of an oval-shaped ceramic aggregate in B6/P4 specimens, whereas the ceramic filler and polymer substrate were homogeneously mixed in B6.7/P3.3. CONCLUSION. Increasing the ceramic filler content improves the mechanical properties, but it can be accompanied by a decrease in the flowability and the homogeneity of the slurry.

• Applied Biological Chemistry
• /
• v.51 no.3
• /
• pp.159-163
• /
• 2008

Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.

• Journal of Life Science
• /
• v.28 no.4
• /
• pp.389-397
• /
• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Communications of the Korean Mathematical Society
• /
• v.23 no.4
• /
• pp.511-528
• /
• 2008

Let (R, m) be a 2-dimensional regular local ring with algebraically closed residue field R/m. Let K be the quotient field of R and $\upsilon$ be a prime divisor of R, i.e., a valuation of K which is birationally dominating R and residually transcendental over R. Zariski showed that there are finitely many simple $\upsilon$-ideals $m\;=\;P_0\;{\supset}\;P_1\;{\supset}\;{\cdots}\;{\supset}\;P_t\;=\;P$ and all the other $\upsilon$-ideals are uniquely factored into a product of those simple ones [17]. Lipman further showed that the predecessor of the smallest simple $\upsilon$-ideal P is either simple or the product of two simple $\upsilon$-ideals. The simple integrally closed ideal P is said to be free for the former and satellite for the later. In this paper we describe the sequence of simple $\upsilon$-ideals when P is satellite of order 3 in terms of the invariant $b_{\upsilon}\;=\;|\upsilon(x)\;-\;\upsilon(y)|$, where $\upsilon$ is the prime divisor associated to P and m = (x, y). Denote $b_{\upsilon}$ by b and let b = 3k + 1 for k = 0, 1, 2. Let $n_i$ be the number of nonmaximal simple $\upsilon$-ideals of order i for i = 1, 2, 3. We show that the numbers $n_{\upsilon}$ = ($n_1$, $n_2$, $n_3$) = (${\lceil}\frac{b+1}{3}{\rceil}$, 1, 1) and that the rank of P is ${\lceil}\frac{b+7}{3}{\rceil}$ = k + 3. We then describe all the $\upsilon$-ideals from m to P as products of those simple $\upsilon$-ideals. In particular, we find the conductor ideal and the $\upsilon$-predecessor of the given ideal P in cases of b = 1, 2 and for b = 3k + 1, 3k + 2, 3k for $k\;{\geq}\;1$. We also find the value semigroup $\upsilon(R)$ of a satellite simple valuation ideal P of order 3 in terms of $b_{\upsilon}$.

• Molecules and Cells
• /
• v.45 no.3
• /
• pp.122-133
• /
• 2022

The aim of this study was to investigating whether lncRNA H19 promotes myocardial fibrosis by suppressing the miR-29a-3p/miR-29b-3p-VEGFA/TGF-β axis. Patients with atrial fibrillation (AF) and healthy volunteers were included in the study, and their biochemical parameters were collected. In addition, pcDNA3.1-H19, si-H19, and miR-29a/b-3p mimic/inhibitor were transfected into cardiac fibroblasts (CFs), and proliferation of CFs was detected by MTT assay. Expression of H19 and miR-29a/b-3p were detected using real-time quantitative polymerase chain reaction, and expression of α-smooth muscle actin (α-SMA), collagen I, collagen II, matrix metalloproteinase-2 (MMP-2), and elastin were measured by western blot analysis. The dual luciferase reporter gene assay was carried out to detect the sponging relationship between H19 and miR-29a/b-3p in CFs. Compared with healthy volunteers, the level of plasma H19 was significantly elevated in patients with AF, while miR-29a-3p and miR-29b-3p were markedly depressed (P < 0.05). Serum expression of lncRNA H19 was negatively correlated with the expression of miR-29a-3p and miR-29b-3p among patients with AF (r_s = -0.337, r_s = -0.236). Moreover, up-regulation of H19 expression and down-regulation of miR-29a/b-3p expression facilitated proliferation and synthesis of extracellular matrix (ECM)-related proteins. SB431542 and si-VEGFA are able to reverse the promotion of miR-29a/b-3p on proliferation of CFs and ECM-related protein synthesis. The findings of the present study suggest that H19 promoted CF proliferation and collagen synthesis by suppressing the miR-29a-3p/miR-29b-3p-VEGFA/TGF-β axis, and provide support for a potential new direction for the treatment of AF.

• Journal of Practical Agriculture & Fisheries Research
• /
• v.20 no.2
• /
• pp.31-38
• /
• 2018

This study was conducted to investigate the possibility of biochar as an alternative medium to peatmoss using for Aster spathulifolius. We cultivated A. spathulifolius in four potting media with different mixing rates (v/v) of peatmoss (P) and biochar (B) as follows: B0+P3, B1+P2, B2+P1, and B3+P0 with vermiculite 3 + perlite 3. Also, we analyzed the chemical properties of media and the plant growth characteristics. The results were as follows: In case of media's chemical condition, B0+P3 and B1+P2 treatments showed higher tendency (p < 0.05). Plant height on B0+P3 and B1+P2 treatments was much higher than that on other treatments (p < 0.05). Root length on B1+P2 treatment was higher than on B0+P3 treatment (p < 0.05). B0+P3 and B1+P2 treatments showed higher number of leaves and dry biomass than other treatments. Therefore, our results support that Biochar : Peatmoss : Vermiculite : Perlite (1/3 : 2/3 : 1 : 1, v/v) could be a more economical potting medium for A. spathulifolius than peatmoss : vermiculite : perlite (1 : 1 : 1, v/v).