• Korean Journal of Microbiology
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• v.23 no.1
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• pp.13-19
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• 1985

Acc I endonuclease has been isolated from 300g (wet weight) cells of Acinetobacter calcoaceticus. The cells were broken by using French press at 20, 000p.s.i. After ammonium sulfate fractionation, the enzyme was further purified by heparin agarose, DEAE-sephades, Affi.-gel Blue, phosphocellulose, and hydroxylapatite column chromatography. The purified Acc I endonudlease has a single polypeptide species and its subunit molecular weight was 45,000 ${\pm}$ 1,000 daltons as judged by 10% SDS-polyacrylamide gel electrophoresis. The isolated enzyme was essentially free of contaminating nucleases as judged by homochromatography by using a $^{32}P-labeled$ oligonucleotide. The enzyme showed maximum activity at pH values between 8.0 and 11.0 and in the presence of $MgCl_2$. Acc I endonuclease was maximally active in the absence of NaCl and was completely inhibited at 200 mM NaCl.

• The Korean Journal of Food And Nutrition
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• v.33 no.3
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• pp.309-316
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• 2020

In this study, we investigated the protective effects of Ulva lactuca methanol extracts against ultraviolet B (UVB)-induced DNA damage in HaCaT cells. First, the contents of general and antioxidative nutrient contents of Ulva lactuca were measured. The moisture, carbohydrate, crude protein, crude fat and ash were 14.01%, 44.80%, 23.19%, 3.10% and 14.90%, respectively. Magnesium that acts as DNA repair enzyme cofactor was the most abundant mineral followed by Ca, P and Fe. The total phenolic and anthocyanoside contents of Ulva lactuca were 2.69 mg/g and 0.13 mg/g, respectively. Cells treated with Ulva lactuca methanol extracts for 24 hours post UVB exposure increased cell viability in a concentration-dependent manner compared to the non-treated control. Also, Ulva lactuca methanol extracts decreased the levels of UVB-induced DNA damage such as cyclobutane pyrimidine dimer and DNA damage response (DDR) proteins such as p-p53 and p21. These results suggest that Ulva lactuca methanol extracts comprising physiological active substances such as Mg, polyphenols and anthocyanosides promote DNA repair by regulating genes related with DDR.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.4
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• pp.547-555
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• 2020

Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (DHCR24) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs. Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and DHCR24-siRNA in vitro. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzyme-linked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and DHCR24 3'-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the DHCR24 mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor. Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of DHCR24 also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within DHCR24 3'-UTR. Up-regulation of miR-26a inhibited DHCR24 mRNA and protein expression in pGCs. Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of DHCR24 in pGCs.

• Reproductive and Developmental Biology
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• v.28 no.4
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• pp.211-219
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• 2004

The effects of adenosine 5'-triphosphate (ATP) and ATP analogs, P/sub 2y/ purinoceptor agonists, on growth of normal mouse mammary epithelial cells (NMuMG) were examined. Cells were plated onto 24 well plates in DMEM supplemented with 10 % fetal calf serum. After serum starvation for 24 hours, ATP, P/sub 2y/ purinoceptor agonists (AdoPP[NH]P, ATP-α-S, ATP-γ-S, β, γ-me-ATP and 2me-S-ATP), P/sub 2u/ purinoceptor agonist (UTP) and P/sub 2y/ purinoceptor antagonists (Reactive Blue 2, more selective to P/sub 2y/ receptor than PPADS; PPADS) were added. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA (1 hour pulse with 1 μ Ci/ml, 18～19 hours after treatment). ATP, Adopp[NH]P, ATP-α-S or ATP-γ-S, significantly increased DNA synthesis at 1, 10 and 100 μM concentrations with dose-dependency (P<0.05), and the maximum responses of ATP and ATP analogs were shown at 100 μM concentration (P<0.05). The potency order of DNA synthesis was ATP≥ATP- γ -S>Adopp [NH]P>ATP-α-S. β, γ -me-ATP, 2me-S-ATP and UTP did not increase DNA synthesis. In autoradiographic analysis of percentage of S-phase cells, similar results were observed to those of DNA synthesis. Addition of 1, 10 or 100 μM Reactive Blue 2 or PPADS significantly decreased ATP (100 μM)-induced DNA synthesis, however, PPADS was less effective than Reactive Blue 2. In Elvax 40P implant experiment, ATP directly stimulated mammary endbud growth in situ suggesting the physiological regulator of ATP in mammary growth. ATP 100 μM rapidly increased MAPK activity, reaching a maximum at 5 min and then gradually decreasing to the base level in 30 min. ATP analogs, Adopp[NH]P and ATP-γ-S also increased MAPK activity, however, β, γ-me-ATP and 2me-S-ATP did not. The inhibitor of the upstream MAPK kinase (MEK), PD 98059 (25 μM), effectively reduced ATP (100 μM) or EGF(10 ng/ml, as positive control)-induced MAPK activity and DNA synthesis (P<0.05). These results indicate that ATP-induced DNA synthesis was prevented from the direct inhibition of MAPK kinase pathway. Overall results support the hypothesis that the stimulatory effects of normal mouse mammary epithelial growth by addition of ATP or ATP analogs are mediated through mammary tissue specific P/sub 2y/ purinoceptor subtype, and MAPK activation is necessary for the ATP-induced cell growth.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.8 no.1
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• pp.93-101
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• 2002

Objectives This experimental study was carried out to evaluate the effects of Phellinus Linteus on immune activation. Methods In order to investigate the effect of Phellinus Linteus, the following was performed: The fraction of $CD4^+$, $CD8^+$, $CD19^+$ in splenic cell, gene expression of IL-12 (p35), IL-12 (p40) , $IFN-{\gamma}$, and splenic cell proliferation by PL-E. Results PL-E helped $CD4^+$, $CD8^+$, $CD19^+$ expression more effectively compared with control group. PL-E helped IL-12 (p35), IL-12 (p40), $IFN-{\gamma}$ gene expression in splenic cells more effectively compared with control group. PL-E proliferated splenic cells more effectively compared with control group. Conclusions: It is suggested that PL-E is able to activate immune response system.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.141-161
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• 2006

Purpose : This study was performed to investigate the effects of Sobokchukeo- Tang(SCT) on the experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced via the surgical autotransplantation technique in rats. A laparotomy was performed and a $4\;{\times}\;4\;mm$ of the right uterine horn was resected incised and sutured to the peritoneum. And the animals divided into control(n=8) and SCT-treated group(n=8). SCT(1,000 mg/head) was administered orally for 15 days after operation. The weights(body, left uterus, and ovaries) and concentrations of cytokines(MCP-1,$TNF-{\alpha}$, $IL-l{\beta}$) were measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells of the transplanted uterine tissues were performed. Results : - The $volume(mm^3)$ of transplanted uterine tissues of SCT-treated group$(92.88{\pm}41.89)$ was significantly(p<<0.01) decreased than the control group $(404.50{\pm}317.68)$. The concentration(pg/ml) of MCP-1 in ascites of SCT-treated group$(5,256{\pm}1,209)$ was significantly(p<<0.001) decreased than the control group$(8,632{\pm}1,245)$. - The concentration(pg/ml) of $TNF-{\alpha}$ in ascites of SCT-treated group$(521.8{\pm}306.1)$ was significantly(p<<0.01) decreased than the control group$(1,245.2{\pm}362.2)$. - The percentage of COX-2 positive epithelial layer in transplanted uterine tissues of SCT-treated group$(25.0{\pm}7.3)$ was significantly(p<<0.001) decreased than the control group$(50.2{\pm}8.2)$. - The number of mast cells in the stroma of transplanted uterine tissues of SCT-treated group$(16.5{\pm}6.8)$ was significantly(p<<0.05) decreased than the control group$(26.0{\pm}7.7)$. - The number of mast cells in the periphery of transplanted uterine tissues of control group$(71.3{\pm}18.5)$ was significantly(p<<0.01) decreased than the control group$(109.3{\pm}30.2)$. - Proliferation of epithelia, infiltration of inflammatory cells, and microanglogenesis in transplanted uterine tissues of treated group were weakly observed than the control group. Conclusion : From the above results, Sobokchukeo-Tang(SCT) has an inhibitory effect on the development of transplanted uterine tissue in rats and it is related to the decreased concentration of MCP-1 and $TNF-{\alpha}$, and decreased expression of COX-2, and decreased infiltration of mast cells by administration of Sobokchukeo-Tang.

• Korean Journal of Acupuncture
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• v.27 no.2
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• pp.95-105
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• 2010

Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.511-519
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• 2009

A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.

• Journal of Environmental Impact Assessment
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• v.19 no.5
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• pp.465-473
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• 2010

This study was carried out to investigate the effect of EFDC hydrodynamic result on the WASP7.3 water quality modeling result in accordance with the change of number of grid for the dam reservoir to be constructed. The simulated flow and BOD, T-N and T-P loads by the HSPF watershed model was used for boundary conditions and the hydrodynamic modeling results was linked with WASP model to predict future water quality after dam construction. The scenarios for EFDC modeling were composed of Scenario 1(141 grid cells) and Scenario 2(568 grid cells). The results of Scenario 2 showed that BOD, T-N, T-P and Chl-a concentrations were decreased 0.073mg/L(8.5%), 0.032mg/L(2.6%), 0.003mg/L(6.8%), 0.644mg/L(4.2%) compared with those of Scenario 1, respectively. As number of grid cell increased, water quality concentrations were decreased and also it caused the longer running time. Therefore, this study suggests that the consideration of the geometry of water body is more important than the number of grid cells for the prediction of water quality of a dam reservoir in EIA.

• Molecules and Cells
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• v.19 no.1
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• pp.143-148
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• 2005

Heptaplatin, cis-malonato [(4R,5R)-4,5-bis (amino-methyl)-2-isopropyl-1,3-dioxolane] platinum(II) (SKI-2053R, Sunpla) is a new platinum derivative with antitumor activity comparable to cisplatin on various cancer cell lines. Preclinical studies suggest that it is less nephrotoxic than cisplatin. This study was undertaken to examine the combined effect of heptaplatin and ionizing radiation on two established human squamous carcinoma cell lines (NCI-H520, SQ20B). The cytotoxic activity of heptaplatin was concentration-dependent in both cell lines. When low dose heptaplatin was combined with high dose ionizing radiation, there was an additive cytotoxic effect on NCI-H520 cells (P < 0.05), while a moderate dose of heptaplatin and a low dose of ionizing radiation had an additive cytotoxic effect on the growth of SQ20B cells (P < 0.05). FACS analysis and DAPI staining showed that their additive cytotoxic effects were correlated with the induction of apoptosis. Further studies are warranted using heptaplatin and ionizing radiation in squamous cell carcinoma as a substitute for cisplatin.