• Korean Journal of Food Science and Technology
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• v.31 no.2
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• pp.553-557
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• 1999

Five fungi (mushrooms), Daedaleopsis styracina, Trichaptum abietium, Coriolus versicolor, Pisolithus tinctorius and Tricholomopsis decora, were screened and examined the fibrinolytic activity and specificity. The extracts of mushrooms showed a level of fibrinolytic activity that was about 3-4 times higher than that of plasmin 1.0 unit. In particular, Pisolithus tinctorius of them showed the greatest enzyme activity (4.71 plasmin unit/mL) by fibrin plate assay, and the highest specificity (1.32 plasmin unit/mL) using chromogenic substrate (N-p-Tosyl-Gly-Pro-Lys p-nitroanilide) by Tricholomopsis decora. And the same molecular mass 54 and 61kDa showing the fibrinolytic activity obtained from all fruiting bodies were confirmed, and it was found that Trichaptum abietium and Tricholomopsis decora have a strong fibrinolytic enzyme with an apparent size of 100 kDa and 84 kDa, respectively on SDS-fibrin zymography activity assay.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.2
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• pp.103-112
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• 2009

Objectives : This study was to evaluate the pharmacological effect of Korea Red Ginseng aqueous extract (KRGE) on serum-deprived apoptosis of neuronal-like pheochromocytoma PC12 cells and to investigate its underlying action mechanism. Methods : KRGE was prepared by extracting Korea Red Ginseng with hot water and concentrating using a vacuum evaporator. Cell viability was determined after incubation of cells with KRGE or chemical inhibitor in serum-deprived medium for 60 h by counting intact nuclei following lysing of the cell membrane. Caspase activities were measured using chromogenic substrates and signal-associated protein phosphorylation and cytochrome c release were determined by Western blot analyses using their specific antibodies. Results : Serum deprivation induced PC12 cell death, which was accompanied by typical morphological features of apoptotic cell, such as nuclear fragmentation, caspase-3 activation, and cytochrome c release. This apoptotic cell death was significantly inhibited by KRGE and caspase-3 inhibitor, but not by the addition of NMA, ODQ, and PD98059. KRGE promoted phosphorylation of Akt and Bad, and this phosphorylation was inhibited by the PI3K inhibitor LY92004. In addition, this inhibitor also reversed KRGE-mediated protection of PC 12 cells from serum deprivation. These results suggested that KRGE protects PC12 cells from serum deprivation-induced apoptosis through the activation of PI3K/Akt-dependent Bad phosphorylation and cytochrome c release, resulting in caspase-3 activation. Conclusions : KRGE should be considered as a potential therapeutic drug for brain diseases including stroke induced by apoptosis of neuronal cells.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.223-229
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• 1988

Bacillus subtilis K-4-3, which produces considerable amount of $\beta$-glucanase was selected among extracellular $\beta$-glucanase-producing bacteria isolated from soil. $\beta$-glucanase was purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-sephacel ion exchange chromatography. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. Its molecular weight was estimated to be 17000 dalton by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature of the purified $\beta$-glucanase were 7.0 and $50^{\circ}C$, respectively. The enzyme was strongly inhibited by 1.0mM of $Fe^{3+}$, and activated by 1.0mm of $Li^{}47+$. The absence of glucose after thin layer chromatography of reaction products revealed that the purified enzyme contains no cellobiase or laminarinbiase activity. The loberation of ki, tri-and tetra-saccharide as reaction products can be explained by endoaction of the enzyme.

• Journal of the Korean Chemical Society
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• v.38 no.7
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• pp.502-508
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• 1994

The measurement of trace amounts of uranium(VI) and thorium(VI) in the solutions containing high concentration of dissolved salts was carried out. The procedure using reversed phase liquid chromatography with trace enrichment techniques has been developed to cope with the high salt content of the samples. 2 ml of sample were passed through a small C_{18}$ reversed phase enrichment column with ${\alpha}$-HIBA eluent (0.11 M, pH 5.5) where the uranium and thorium were separated from other constituents and concentrated. The uranium and thorium were then backflushed from the column onto a deactivated C_{18}$ reversed phase analytical column where furthe separation was achieved with a mixed eluent (pH3.0, 0.17M ${\alpha}$-HIBA/0.0038 M 1-pentanesulfonate). The separated species were determined spectrophotometrically by postcolumn reaction with Arsenazo III, the chromogenic reagent. Detection limits were found within 1 ppb range for both species.

• BMB Reports
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• v.29 no.6
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• pp.500-506
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• 1996

We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.

• Food Science of Animal Resources
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• v.27 no.4
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• pp.496-500
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• 2007

The lactase activities of nine species of lactic acid bacteria were compared using the chromogenic substrate, $o-nitrophenyl-{\beta}-D-galactopyranoside$. Streptococcus thermophilus KACC 91147 had the highest lactase activity among a total of thirty strains of Lactobacillus and S. thermophilus tested, including commercial strains. S. thermophilus KACC 91147 released $0.30{\pm}0.12\;mg/mL$ of galactose in treated milk A ($10^7\;CFU/mL$) and $6.49{\pm}0.38\;mg/mL$ in treated milk B ($10^9\;CFU/mL$ milk) over 2 hours. In milk tolerance tests, the blood glucose level (BGL) of 6 volunteers (2 males and 4 females) clinically diagnosed as lactose intolerant increased 3.0 mg/dl after drinking milk A, but a significant (p<0.05) additional increase of $11.2{\pm}4.18\;mg/dl$ was found after drinking milk B. This result suggests that the addition of S. thermophilus KACC 91147 cells into milk aids the digestion of lactose in milk and ameliorates the symptoms of lactose-intolerant individuals due to the activity of lactase from the lactic streptococci.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.235-242
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• 2000

The purpose of this in vitro study was to evaluate the sealing ability of three sealers(Sealapex, Pulp canal sealer, AH26) used with continuous wave method using an anaerobic bacterial leakage model. 53 extracted human teeth with straight and single canals were prepared with crown-down pressureless technique using .04, .06 taper Profile(Maillefer, Swiss). Master apical file was maintained as #35 K-file. All canals of the experimental teeth were obturated with continuous wave method using System B(Analytic technology, U.S.A.) The teeth were randomly divided into three experimental groups of 15 and two control groups of 4. Experimental group 1 was obturated with Sealapex and group 2 with Pulp canal sealer, and group 3 with AH26. A dual chamber anaerobic bacterial leakage model was assembled. Brain heart infusion with yeast extract, hemin, menadion, and the chromogenic indicator bromocresol purple was used as the culture broth for Fusobacterium nucleatum(VPI 10197), The specimens were incubated in anaerobic chamber at $37^{\circ}C$ and were observed every 2 to 3 clays, The coronal leakage was evaluated through the color change of culture broth in lower chamber for 60 days. The results were as follows: 1. The incidence of bacterial leakage in group 1 (Sealapex group was 80%, 53% in group 2 (Pulp canal sealer), 27% in group 3 (AH26). 2. There were statistically significant differences in leakage scores between group 1 and group 2, and between group 1 and group 3, respectively. (P<0.05) 3. There was no significantly difference in leakage score between group 2 and group 3. (P>0 05)

• Journal of Oral Medicine and Pain
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• v.46 no.2
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• pp.33-40
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• 2021

Purpose: Oral candidiasis is the most common fungal infection in the oral cavity which is usually diagnosed from clinical findings. A retrospective study was conducted to identify risk factors for oral candidiasis and to characterize the demographic and clinical features of affected patients. Methods: From January 1, 2019 to December 31, 2019, it consisted of 90 oral candidiasis patients diagnosed based on clinical finding and treated with antifungal drugs. As a retrospective study of those people, surveys were conducted on sex, age, systemic disease, a use of dentures, complaints of dry mouth, smoking and alcohol consumption, culture on potato dextrose agar (PDA) medium, culture on chromogenic agar (CA) medium and a duration of antifungal treatment. Results: Among 90 selected patients, the male and female ratio was 41:49. Overall, female had a higher infection rate than male in all age groups. In this study, oral candidiasis was not clearly susceptible to dry mouth, smoking or drinking, wearing dentures and association with systemic disease. Among 90 patients with oral candidiasis, 83 had colonies formed on PDA medium and 53 had colonies formed on CA medium. The duration of antifungal treatment was highest between 5 and 8 weeks. In addition, there was statistical significance between the culture results in CA medium and the duration of antifungal treatment. Conclusions: Generally, old age or infants, dry mouth, smoking, a use of dentures and endocrine abnormalities are risk factors to increase oral candidiasis; however, in this study, it was mainly found in the elderly aged 60 or older regardless of sex and the incidence of oral candidiasis was not obviously related with patients with dry mouth, smoking or drinking, denture wearers and endocrine abnormalities. Interestingly, when the fungi were cultured in CA medium, the duration of antifungal treatment was increased.

• Analytical Science and Technology
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• v.20 no.4
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• pp.339-346
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• 2007

To search the ninhydrin derivatives that have high chromogenic and fluorogenic properties, molecular holographic quantitative structure property relationship (HQSPR) models on the reactivity between glycine and ninhydrin analogues as latent fingerprint detector were derived and investigated quantitatively. The ${\varepsilon}LUMO$ (e.v.) energy of ninhydrin molecule was an important factor to reactivity of ninhydrin. And, it is suggested that the nucleophilic reaction by orbital-controlled reaction from the frontier molecular orbital (FMO) interaction between glycine and ninhydrin derivatives was more superior than that of electrophilic reaction by charged controlled reaction. The analytical results in atomic contribution maps also shows that the reactivity of ninhydrin was increased by meta-substituents as strong electron withdrawing groups on the benzo ring. Therefore, it is sugested by HQSPR and QSPR model that the 5,6-dinitroninhydrin molecule would increase the reactivity as much as three times as compared to none substituted ninhydrin molecule.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1271-1283
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• 2007

A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.