• Journal of Life Science
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• v.28 no.1
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• pp.99-104
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• 2018

Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

• Journal of Dairy Science and Biotechnology
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• v.38 no.2
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• pp.89-98
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• 2020

The purpose of this study was to investigate the probiotic properties of lactic acid bacteria (LAB) isolated from a Korean traditional food kimchi. Gram staining was performed by Macrogen (Macrogen, Inc.) for identification of the LAB. Five strains of LAB were identified, including DKGF9 (Lactobacillus plantarum), DKGF1 (L. paracasei ), DKGF8 (L. casei ), DK207 (L. casei ), and DK211 (L. casei ). The biological activities of the isolated strains were assessed. The results showed that heat resistance of the strains was similar to or higher than the commercial strain L. acidophilus LA-5. Indirect testing of the ability of the strains to attach to the mucin layer revealed that DKGF9, DKGF1, and DKGF8 have high binding affinities for the mucous layer. All strains showed antimicrobial activity similar to or higher than the commercial strain LA-5. In proteolysis experiments, the diameters of proteolysis zones of the five strains increased in the period of 24-72 h, with DKGF1 exhibiting the largest zone diameter. Three strains were selected based on their antioxidant activities. Among the five isolated strains, L. paracasei DKGF1 showed potential probiotic activity, and thus, it may be useful for the development of health-promoting products.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.671-675
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• 2002

The present study was conducted to elucidate functional properties of sulfated polysaccharides from ascidian tunics, In physical properties of the crude polysaccharides, emulsion ability and foaminess were more excellent compared with chitin and chitosan, particular dye binding capacity was prominent. Anti-blood coagulation of partially purified sulfnted polysaccharides showed with respect to APTT (Activated partial thromboplastin time). Especially, active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed highest activity, which was approximately $20\%$ of the activity of heparin. ACE inhibitory activity also similar to anticoagulant activity. Active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed about $34\%$, in ACE inhibitory activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.4
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• pp.460-465
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• 2005

Water and ethanol extracts from Rue were prepared, and their growth inhibiting activity against Helicobacter pylori and other biological activities were examined. Total phenolic compounds in the water and ethanol extracts were present at the concentration of 16.39 mg/g, and 17.07 mg/g, respectively. At the concentration of $200\;{\mu}g/mL$ of phenolic compounds concentration, water extract produced 12 mm inhibition zone while ethanol extract produced 13 mm. The ABTS [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] radical decolorization and antioxidant protection factor (PF) were determined for extracts from Rue. Water extracts showed $96\%$ inhibition rate on ABTS, but ethanol extracts showed higher PF (1.2) than water extracts (0.8). Water extracts had higher electron donation ability on DPPH than ethanol extracts. But ethanol extracts had higher ACE (angiotensin converting enzyme) and xanthine oxidase inhibitory activities than water extracts. Rosemarinic acid and quercetin were the most abundant phenolic compounds as analyzed by HPLC.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.9
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• pp.1308-1313
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• 2005

The physiological activity of Lycii fructus extracts were examined. Total phenolic contents in the ethanol extracts (9.5 mg/g) of Lycii fructus were higher than that of water extracts (8.7 mg/g). The chlorogenic acid ($1.7{\mu}g$ in water extracts and $1.3{\mu}g$ in $60\%$ ethanol extract) was the most abundant phenolic compound as analyzed by HPLC. The ABTS [2,2'-azinobis (3-othylbenfothiaznoline-6- sulfornic acid)] radical decolor-ization electron donating ability (DPPH) and antioxidant protection factor (PF) were determined for extracts from Lycii fructus. Water extract ($76.7\%$ on ABTS, $92.6\%$ on DPPH and 1.1 on PF) showed higher inhibition rate than $60\%$ ethanol extracts ($52.8\%,\;88.8\%$ and 1.0). Thiobarbituric acid reactive substance (TBARS) was determined as $1.5{\times}10^{-3}\;{\mu}M$ in $60\%$ ethanol extract. Ethanol extracts was more effective in decreasing TBARS than water extracts. The water extracts from Lycii fructus had higher angiotensin converting enzyme (ACE) inhibition activity than ethanol extracts. The result will be useful for functional foods application and under-standing the physiological activities of Lycii fructus.

• Applied Biological Chemistry
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• v.49 no.4
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• pp.298-303
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• 2006

The biological and antioxidative activity of Phellinus linteus extracts from gradient ethanol concentrations were examined. The phenol contents of Phellinus linteus(28.36 mg/100 ml) was higher in the 80% ethanol extracts than other extracts. Electron donation ability on DPPH of 80% and 90% ethanol extracts(94.12% and 94.14% inhibition) from Phellinus linteus were the highest. The antioxidant activity against water soluble materials of Phellinus linteus ethanol extracts showed totally high inhibition rates above 80%, especially in 80% and 90% ethanol extracts, they showed each 94.12% inhibition and 94.14% inhibition. The inhibition against ABTS [2,2azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] radical decolorization of 80% ethanol extracts was the highest as 96.2%. The antioxidant protection factor (PF) against lipid soluble materials was the highest in 80% ethanol extracts as 1.69 PF, and TBARS of 80% and 90% ethanol extracts were lower as $1.15{\times}100{\mu}M$ and $1.21{\times}100{\mu}M$ than control($1.95{\times}100{\mu}M$. Angiotensin converting enzyme and xanthine oxidase inhibitory activity of 80% ethanol extracts from Phellinus linteus was higher as 95.10%, 85.07% than other extracts. The results to analized of simple phenolic compounds of Phellinus linteus ethanol extrcts with HPLC showed that they were procatecuic acid, caffeic acid and coumaric acid.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1391-1398
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• 2005

The anti-thrombic properties of the oriental herbal medicine Daejowhan(DJW, 大造丸) which consists of 11 kinds of herbs (indicated as ratio) of Rehmanniae Radix 24%, Hominis Placenta 5%, Testudinis Carapax 9%, Eucommiae Cortex 9%, Asparagi Radix 9%, Phellodendri Cortex 9%, Achyranthis Radix 7%, Liriopis Tuber 7%, Angelicae Sinensis Radix 7%, Ginseng Radix 5% and Schizandrae Fructus 3% were investigated. The water extracts from DJW inhibited Platelet-activating factor(PAF) induced platelet aggregation. DJW was extracted with methanol and further fractionated by ethylacetate. A 70% methanol extract showed a strong inhibition against PAF-induced aggregation in vitro and in vivo assays. The ethylacetate soluble fraction was shown to have inhibitory effect on PAF-induced platelet aggregation in vitro assay. The ethylacetate soluble fraction specially protected against the lethality of PAF, while verapamil did not afford any protection. These results indicate that the water extracts and alcoholic-fractions inhibit the action of PAF in vivo by an antagonistic effect on PAF, so that it may be useful in treating disorders caused by PAF, such as acute allergy, inflammation, asthma, gastrointestinal ulceration, toxic shock and so forth. DJW was investigated regarding its assumed anti-thrombic action on human platelets which was deduced from its ability to suppress Arachidonic acid(AA)-induced aggregation, exocytosis of ATP, and inhibition of Cyclooxygenase(COX) and Thromboxane synthase(TXS) activity. The latter two effects were estimated from the generation of Prostaglandin $E_2(PGE_2)$ and Thromboxane $A_2(TXA_2)$ respectively. Exogenously applied AA ($100{\mu}mol/{\ell}$) provoked a $89\%$ aggregation of platelets, the release of 14 pmol ATP, and the formation of either 225 pg $TXA_2$ or 45 pg $PGE_2$, each parameter being related to 106 platelets. An application of DJW 5 min before AA dose-dependently diminished aggregation, ATP-release and the synthesis of $TXA_2$ and $PGE_2$ with $IC_{50}$ values of 74, 108, 65, $72{\mu}g/m{\ell}$, respectively. The similarity of the $IC_{50}$ values suggest an inhibition of COX by DJW as primary target, thus suppressing the generation of $TXA_2$ which induces aggregation of platelets and exocytosis of ATP by its binding on $TXA_2$-receptors.

• Korean Journal of Community Nutrition
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• v.6 no.5
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• pp.726-733
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• 2001

This study was designed to investigate the effect of iron supplementation on the iron nutritional status and anemia of high school girls in Korea. One hundred thirty-five female students residing in Ulian metropolitan city in Korea diagnosed as having anemia or iron deficiency participated in this study. One or two tablets of iron medicine(80-160 mg Fe as ferrous sulfate/day) were administered to all participants for 3 months. Subjects were evaluated with a questionaire, measurement of hematological indices before and after iron supplementation. The average height and weight of respondents were 161.62 $\pm$ 4.68 cm and 53.87 $\pm$ 6.10 kg, respectively. Daily intakes of energy were 1597.8 $\pm$ 302.35 kcal(76.0% RDA). Iron intakes were 13.72 $\pm$ 4.17 mg (76.3% of RDA) and calcium intakes were 580.74 $\pm$ 177.21(72.5% of RDA) before iron supp]ementation. At baseline, 63% of all participants had depleted store(serum ferritin 12 ug/ml and/or transferrin saturation(TS) < 14%). After iron supplementation, this proportion declined to 19.3%. 55.6% of subjects had 12 ug/m1 of basal ferritin concentration before iron supplementation, and this proportion declined to 16.3% after iron supplementation. The basal hemoglobin(Hb) concentrations were 12.13 $\pm$ 1.01 g/dl and they increased to 12.79 $\pm$ 0.81 g/dl, which showed significant difference artier iron supplementation(p < 0.001). The basal ferritin and TS(%) were 13.24 $\pm$ 11.66 ng/ml, 18.42 $\pm$ 10.12% and they significantly increased to 32.95 $\pm$ 21.14 ng/ml, 33.53 $\pm$ 16.64%, respectively(p < 0.001). The basal total iron binding protein(TIBC) were 467.81 $\pm$ 97.24 ug/dl and they significantly decreased to 325.05 $\pm$ 48.89 ug/dl(p < 0.001) after iron supplementation. The number of tablets administered was positively correlated with serum iron(t = 0.553, p < 0.01), serum ferritin(t = 0.557, p < 0.01), TS(%)(t = 0.588, p < 0.01) and negatively correlated with TIBC(t= -0.409, p <0.01). The anemia symptoms such as ‘Shortening of breath when going upstairs(p < 0.01)’, ‘Tired out easily(p < 0.01)’, ‘Feeling blue(p < 0.001)’, ‘Decreased ability to concentrate(p < 0.01)’, and ‘Poor memory(p < 0.001)’improved significantly after iron supplementation. In this study, daily iron supplementations were efficacious in improving the iron status and anemic symptoms of female high school students. Regular check-ups and nutrition education for adolescents are necessary because of their vulnerability to iron deficiency. Further studies are needed to determine the minimum effective dose of iron and to examine the adverse effect of long-term iron supplementation.

• Food Science and Preservation
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• v.24 no.7
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• pp.885-890
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• 2017

For the construction of the microbial monitoring method, anti-Salmonella polyclonal antibodies (pAbs) were produced from a rabbit and purified by saturated ammonium sulfate precipitation and protein A affinity column. The reactivity of anti-Salmonella pAbs was compared to that of commercial ones by using an indirect ELISA. The specificity of anti-Salmonella pAbs was investigated using 20 Salmonella serotypes and 20 non-Salmonella strains. A capturing ability of anti-Salmonella pAbs was investigated by exposing antibody-immobilized gold biosensor to different concentration of Salmonella mixture. Anti-Salmonella pAbs were successfully produced and purified with an antibody concentration of 2.0 mg/mL The reactivity of purified anti-Salmonella pAbs was greater than that of commercial one at all tested concentrations. All Salmonella serotypes, except S. Diarizonae, showed excellent binding efficiency with purified anti-Salmonella pAbs. Moreover, the purified anti-Salmonella pAbs showed excellent specificity against all non-Salmonella strains. The anti-Salmonella pAbs immobilized on the gold biosensor demonstrated the successful capturing capability against Salmonella with a dose-response manner. Therefore, the anti-Salmonella pAbs exhibited sufficient reactivity, specificity, as well as capturing capability against Salmonella to be considered as a bio-recognition element.

• BMB Reports
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• v.33 no.6
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• pp.454-462
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• 2000

Interleukin-4(IL-4) is known to be a major cytokine regulating immunoglobulin E(IgE) response by the induction of IgE production and type II IgE receptor(IgER II: CD23) expression. Recently, however, the role of neuroendocrine factors has been implicated in modulating the IgE response. Among various neuroendocrine growth factors, we investigated the effects of the insulin-like growth factor-1(IGF-1) since IL-4 and IGF-1 share common intracellular signaling molecules, such as the insulin receptor substrate-1/2(IRS-1/2) to induce a specific cellular response. In the human peripheral blood mononuclear cell (PBMC) cultures, IGF-1 was capable of inducing a substantial level of IgE production in a dose-dependent manner. It also noticeably upregulated the IL-4-induced or IL-4 plus anti-CD40-induced IgE production. Similarly, the IGF-1-induced IgE production was enhanced by IL-4 or anti-CD40 in an additive manner, which became saturated at high concentrations of IGF-1. Although IGF-1 alone did not induce IgER II (CD23) expression, it augmented the IL-4-induced surface CD23 expression in a manner similar to the action of anti-CD40. These results imply that IGF-1 is likely to utilize common signaling pathways with IL-4 and anti-CD40 to induce IgE and IgER II expression. In support of this notion, we observed that IGF-1 enhanced the IL-4-induced signal transducers and activators of transcription 6(STAT6) activation and independently induced $NF-{\kappa}B$ activation. Both of these bind to the IgE(C) or IgER II (CD23) promoters. Together, our data suggest that IL-4 and IGF-1 work cooperatively to activate STAT6 and $NF-{\kappa}B$. This leads to the subsequent binding of these transcription factors to the $C{\varepsilon}$ and CD23 promoters to enhance the expression of IgE and IgER II. The observed differential ability of IGF-1 on the induction of IgE vs. IgER II is discussed based on the different structure of the two promoters.