• Biomolecules & Therapeutics
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• 제20권1호
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• pp.81-88
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• 2012

Histone deacetylases (HDACs) are enzymes involved in the remodelling of chromatin, and have a key role in the epigenetic regulation of gene expression. Histone deacetylase (HDAC) inhibitors are emerging as an exciting new class of potential anti-cancer agents. In recent years, a number of structurally diverse HDAC inhibitors have been identifi ed and these HDAC inhibitors induce growth arrest, differentiation and/or apoptosis of cancer cells in vitro and in vivo. However, the underlying molecular mechanisms remain unclear. This study aimed at investigating the anti-tumor activity of various HDAC inhibitors, IN-2001, using T47D human breast cancer cells. Moreover, the possible mechanism by which HDAC inhibitors exhibit anti-tumor activity was also explored. In estrogen receptor positive T47D cells, IN-2001, HDAC inhibitor showed anti-proliferative effects in dose-and time-dependent manner. In T47D human breast cancer cells showed anti-tumor activity of IN-2001 and the growth inhibitory effects of IN-2001 were related to the cell cycle arrest and induction of apoptosis. Flow cytometry studies revealed that IN-2001 showed accumulation of cells at $G_2$/M phase. At the same time, IN-2001 treatment time-dependently increased sub-$G_1$ population, representing apoptotic cells. IN-2001-mediated cell cycle arrest was associated with induction of cdk inhibitor expression. In T47D cells, IN-2001 as well as other HDAC inhibitors treatment significantly increased $p21^{WAF1}$ and $p27^{KIP1}$ expression. In addition, thymidylate synthase, an essential enzyme for DNA replication and repair, was down-regulated by IN-2001 and other HDAC inhibitors in the T47D human breast cancer cells. In summary, IN-2001 with a higher potency than other HDAC inhibitors induced growth inhibition, cell cycle arrest, and eventual apoptosis in human breast cancer possibly through modulation of cell cycle and apoptosis regulatory proteins, such as cdk inhibitors, cyclins, and thymidylate synthase.

• 한국식품저장유통학회지
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• 제24권4호
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• pp.524-528
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• 2017

Ricinus communis, belongs to the family Euphorbiaceae, has been known as medicinal plants for treatment of inflammation, tumors, antidiabetic, hepatoprotective and laxative. Compared to many pharmacological studies, the effect of R. communis extract on regulating adipogenesis as therapeutic drug for treating obesity has not been reported. R. communis extract (RCE) was investigated to determine its effects on the adipogenesis by monitoring the status of $Wnt/{\beta}-catenin$ signaling and factors involving the differentiation of adipocytes. The differentiation of 3T3-L1 cells monitored by Oil Red O staining was inhibited in concentration dependent manner by RCE. The luciferase activity of HEK 293-TOP cells containing pTOPFlash with Tcf4 response element-luciferase gene was increased approximately 2-folds by the treatment of RCE at concentrations of $100{\mu}g/mL$ compared to the control. Activation of the $Wnt/{\beta}-catenin$ pathway by RCE was further confirmed by immunocytochemical analysis which shows an increment of nuclear localization of ${\beta}-catenin$. In addition, safety of RCE was verified through performing neural stem cell morphology assay. Among the identified flavonoids in RCE, isoquercitrin was the most abundant. Therefore, these results indicate that the adipocyte differentiation was significantly reduced by isoquercitrin in R. communis. In this study, RCE suppresses the adipogenesis of 3T3-L1 cells via the activation of $Wnt/{\beta}-catenin$ signaling.

• Molecules and Cells
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• 제20권2호
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• pp.288-296
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• 2005

Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.

• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1551-1558
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• 2014

The esterase gene Est8 from the thermophilic bacterium Bacillus sp. K91 was cloned and expressed in Escherichia coli. The monomeric enzyme exhibited a theoretical molecular mass of 24.5 kDa and an optimal activity around $50^{\circ}C$ at pH 9.0. A model of Est8 was constructed using a hypothetical YxiM precursor structure (2O14_A) from Bacillus subtilis as template. The structure showed an ${\alpha}/{\beta}$-hydrolase fold and indicated the presence of a typical catalytic triad consisting of Ser-11, Asp-182, and His-185, which were investigated by site-directed replacements coupled with kinetic characterization. Asp-182 and His-185 residues were more critical than the Ser-11 residue in the catalytic activity of Est8. A comparison of the amino acid sequence showed that Est8 could be grouped into the GDSL family and further classified as an SGNH hydrolase. Est8 is a new member of the SGNH hydrolase subfamily and may employ a different catalytic mechanism.

• Environmental Analysis Health and Toxicology
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• 제20권1호
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• pp.13-21
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• 2005

The purpose of this study was to determine the effects of bisphenol A (BPA), which is known to have estrogenic activity, on the early development of medaka fish (Oryzias latipes). The fertilized eggs of medaka were treated with BPA at different concentrations for 3 weeks. Embryonic growth, deformation, hatching success, and gonadal differentiation were determined to observe the effects of this chemical. Also we tried to measure the estrogenic activity of bisphenol A using ELISA and RT-PCR methods. By using this techniques, we evaluated the induction of vitellogenin, an estrogen-regulated gene from the whole body-homogenates of larvae. At results, a reduced blood circulation was seen in embryos and peritoneal edema and hindrance of yolk-sac absorption were observed in larvae of treated group. However, BPA at the concentrations tested (2～200 ㎍/L) did not have severe adverse effects on the early life-stages. According to the observation of gonadal histology, inter-sex or sex -reversal was not found in all test fish. After the exposure was ended, vitellogenin mRNA and protein levels were measured in larvae and then their levels were found to be increased in treated group with 200㎍/L. These results indicate that BPA can induce the expression of vitellogenin in early life-stages as well as in adult male fish.

• 동국한의학연구소논문집
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• 제8권1호
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• pp.117-131
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• 1999

Yangkyuksanhoa-tang is frequently used for cerebrovascular accident(CVA). The present study was performed to investigate the effect of Yangkyuksanhoa-tang on the peri vascular immunoreactive nerve fiber of the basilar artery after experimentally induced subarachnoid hemorrhage(SAH). Sprague Dawley rats weighing between 350-400g were used. The SAH induced by injection of the fresh autologus heart blood(0.3-0.4ml) into the cisterna magna through the posterior atlanto-occipital membrane. Sample group was given a $3.3m{\ell}/kg/day$ of Yangkyuksanhoa-tang extracts for 2 days after SAH. The experimental animals divided into 48hrs after SAH. The changes of perivascular immunoreactive nerve fiber was examined by using indirect immunofluorescence method. The meshlike perivascular nerve fiber appeared in the basilar artery of normal rats. In basilar artery of SAH elicitated rat, the distribution of calcitonin gene-related peptide (CGRP)-immunoreactivity(IR) and vasoactive intestinal polypeptide(VIP)-IR of the perivascular nerve fiber were remarkably diminished, also dopamine beta hydroxylase(DBH)-IR, neuropeptide Y(NPY)-IR and serotonin-IR were diminished. In SAH elicitated rat with Yangkyuksanhoa-tang treatment, the CGRP-IR and VIP-IR degree were repaired as well as normal rat's, but DBH-IR, NPY-IR and serotonin-IR had no changes. These results provide the basic data to investigate the effect of Yangkyuksanhoa-tang on the vasospasm after SAH.

• Journal of Microbiology
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• 제39권2호
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• pp.95-101
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• 2001

In an effort to develop an efficient expression and secretion system for heterologous proteins in Aspergilius nidulans, the PCR-amplified coding sequence for alkaline pretense (AlpA) of A. oryzae was cloned into a fungal expression vector downstream of A. nidulans aicA (alcohol dehydrogenase) promoter to yield pRAAlp. Transformation of A. nidulans with pRAAlp gave stable transformants harboring various copy numbers (3 to 10) of integrated alpA gene, from among which 6 representatives were selected. On a medium containing 0.8% ammonium sulfate that represses the expression of the host's own pretense, the alcA prumoter-controlled AlpA expression was strongly induced by threonine but repressed by glucose. The level of AlpA secretion was highest (approximately 666 mU/ml) in transformant ALP6 containing the largest copy number integrated alpA. However, the level of AlpA secretion was not necessarily proportional to the copy numbers of the integrated alpA genes. The N-terminal sequence or the secreted mature AlpA was determined to be Gly-Leu-Thr-Thr-Gln-Lys-Ser and its molecular mass to be approximately 34 kDa, indicating that AlpA is properly processed by the removal of 121 N-terminal amino acids.

• KSBB Journal
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• 제18권4호
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• pp.294-300
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• 2003

본 연구에서는 L. fermentum KLB12을 열 전처리 함으로서 제제화 과정 동안 거치게 되는 열 스트레스에 대한 내성이 증진됨을 확인하고, 최적의 열 전처리 조건을 수행하였다. 또한 열 전처리 뿐만 아니라, 저온과 열 전처리 조건에도 열 스트레스에 대한 간섭 효과를 확인하였다. 그리고 내성 증진에 신규 단백질 합성이 필요함을 확인하였으며 나아가, 2-D electrophoresis를 통하여 7개의 신규 단백질을 확인하였다. 따라서 이 균주를 제제화하기 위한 방법으로 열 전처리를 이용할 경우 생균력 유지에 큰 효과를 얻을 수 있다고 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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• pp.319-325
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• 2015

Among solute carrier proteins, the organic anion transporters (OATs) play an important role for the elimination or reabsorption of endogenous and exogenous negatively charged anionic compounds. Among OATs, SLC22A9 (hOAT7) transports estrone sulfate with high affinity. The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss. The present study was performed to search the SNP within exon regions of SLC22A9 in Korean females with osteoporosis. Fifty healthy controls and 50 osteoporosis patients were screened for the genetic polymorphism in the coding region of SLC22A9 using GC-clamped PCR and denaturing gradient gel electrophoresis (DGGE). Six SNPs were found on the SLC22A9 gene from Korean women with/without osteoporosis. The SNPs were located as follows: two SNPs in the osteoporosis group (A645G and T1277C), three SNPs in the control group (G1449T, C1467T and C1487T) and one SNP in both the osteoporosis and control groups (G767A). The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively. The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and $12.74{\mu}M$, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively. The present study demonstrates that the SLC22A9 variant F426S is causing inter-individual variation that is leading to the differences in transport of the steroid sulfate conjugate (estrone sulfate) and, therefore this could be used as a marker for certain disease including osteoporosis.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1446-1454
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• 2021

Herein, we cloned and expressed an endo-β-1,4-glucanase gene (celA1805) from Bacillus subtilis B111 in Escherichia coli. The recombinant celA1805 contains a glycosyl hydrolase (GH) family 8 domain and shared 76.8% identity with endo-1,4-β-glucanase from Bacillus sp. KSM-330. Results showed that the optimal pH and temperature of celA1805 were 6.0 and 50℃, respectively, and it was stable at pH 3-9 and temperature ≤50℃. Metal ions slightly affected enzyme activity, but chemical agents generally inhibited enzyme activity. Moreover, celA1805 showed a wide substrate specificity to CMC, barley β-glucan, lichenin, chitosan, PASC and avicel. The K_m and V_max values of celA1805 were 1.78 mg/ml and 50.09 µmol/min/mg. When incubated with cellooligosaccharides ranging from cellotriose to cellopentose, celA1805 mainly hydrolyzed cellotetrose (G4) and cellopentose (G5) to cellose (G2) and cellotriose (G3), but hardly hydrolyzed cellotriose. The concentrations of reducing sugars saccharified by celA1805 from wheat straw, rape straw, rice straw, peanut straw, and corn straw were increased by 0.21, 0.51, 0.26, 0.36, and 0.66 mg/ml, respectively. The results obtained in this study suggest potential applications of celA1805 in biomass saccharification.