• Biomedical Science Letters
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• v.11 no.4
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• pp.509-515
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• 2005

This study was conducted to determine the kinetics of cyclohexane metabolites (the biomarker on cyclohexane exposure), the changes of hepatic cyclohexane metabolizing enzyme activities and the metabolites of cyclohexane in urine or serum. The rats were sacrificed at 2, 4, 8, 12 and 24 hr after administration of one dose of cyclohexane (1.56 g/kg body weight, i.p.). The metabolites of cyclohexane in urine were identified as cyclohexanol, cyclohexanone, trans-l,2-cyclohexanediol and 1,4-cyclohexanediol with cyclohexane metabolite being 124.00, 0.78, 23.28 and 2.75 (g/g of creatinine, $1\times10^{-3}$). Most of the cyclohexanol and trans-l,2-cyclohexanediol were determined to be in the form of $\beta-glucuronide$ conjugates, whereas cyclohexanone and 1 ,4-cyclohexanediol were found as free forms. In toxicokinetics of serum cyclohexane metabolites, cyclohexanol showed a rapid increase, reaching the plateau at 4 hr, after this time rapidly decreased throughout 24 hr. Changes of cyclohexanone also showed the similar pattern with cyclohexanol except somewhat lower concentration. Trans-l,2-cyclohexanediol, however, showed a gradual increase until 12 hr with the continued same levels throughout 24 hr. On the other hand, 1,4-cyclohexanediol was detected as trace levels at 4 and 12 hr, respectively. The administration of cyclohexane led to a significant increase of hepatic aniline hydroxylase activity from 2 to 8 hr. The activity of hepatic alcohol dehydrogenase showed a significant increase at 4 hr and then were recovered to the level of the control at 24 hr. On the other hand, there were no differences in liver weightlbody weight between the control and cyclohexane-treated animals. However, there were the changes of aniline hydroxylase and alcohol dehydrogenase activities on time-dependent pattern after cyclohexane treatment, which influence on the degree of cyclohexane metabolites both in blood and urine. These results suggest that differential determination of cyclohexane metabolites in urine and serum may be able to be as a biomarker of cyclohexane-exposure in the body. But in this fields further study is needed.

• Journal of Pharmaceutical Investigation
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• v.22 no.3
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• pp.173-183
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• 1992

To study the feasibility of transmucosal delivery of $[D-ala^2]-methionine$ enkephalinamide (YAGFM), its enzymatic degradation and stabilization in various rabbit mucosal extracts were investigated by HPLC method. The degradation of YAGFM was observed to follow the first-order kinetics and the half-lives of YAGFM in the nasal, rectal and vaginal mucosal extracts were found to be 25.7, 3.0 and 7.8 hr, respectively. However, there was no significant difference in degradation rates of YAGFM between the mucosal and serosal extracts obtained from the same mucosal membrane. This finding suggests that even a synthetic enkephalin analog, which is designed to be resistent to aminopeptidases, needs to be fully protected from the enzymatic degradation in mucosal sites for the delivery of the analog through mucosal routes. To inhibit the degradation of YAGFM in various mucosal extracts, effects of enzyme inhibitors such as bestatin (BS), amastatin (AM), thiorphan (TP), thimerosal (TM) and EDTA, alone or in combination, and modified cyclodextrins were observed by assaying YAGFM staying intact during 24 hr-incubation at $37^{\circ}C$. It was found from the results that mixed inhibitors such as TM (0.5 mM)/EDTA (5 mM) or AM $(50{\mu}M)/TM$ (0.5 mM)/EDTA (5 mM) provided very useful means for the stabilization in various mucosal extracts. The latter was found to protect YAGFM from the degradation in the nasal, rectal, and vaginal mucosal extracts by 90.9, 90.4 and 91.3%, respectively, after 24 hr-incubation, suggesting almost complete inhibition of YAGFM-degrading enzymes present in the incubation mixture. However, BS $(50{\mu}M)$, AM 50 $(50{\mu}M)$ or TP$(50{\mu}M)$ alone did not reveal sufficient inhibition except TM (0.5 mM) or EDTA (5 mM). The adddition of $2-hydroxylpropyl-{\beta}-cyclodextrin$(10%) to the nasal mucosal extract, and $dimethyl-{\beta}-cyclodextrin$(10%) to the rectal and vaginal mucosal extracts reduced the first-order rate constants for the degradation of YAGFM by 5.8, 17.3 and 8.9 times, respectively, compared to those with no additive.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.3 s.52
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• pp.273-277
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• 2005

This study was performed to develop new cosmeceutical agents with sebosuppressive activity from native plant extracts in Korea. Inhibitory efforts of the extracts on $5{\alpha}-reductase$ (5-AR) were evaluated by enzyme kinetics analysis using UV-spectrophotometric method. Two kinds of enzyme suspensions as 5-AR sources were prepared from rat liver tissue and cultured hSG cells. The sebosuppressive effects were determined by measuring the total lipid quantity produced in cultured hSG cells after incubation with the extracts. As a result, Pinus thunbergii extracts showed the most potent 5-AR inhibitory effects. Its $K_i$ values were 0.0002% and 0.0014% for rat liver 5-AR and human sebaceous gland 5-AR, respectively. Addition of Pinus thunberii extract to hSG cells showed 48% reduction in total lipid production at 0.005% concentration. In conclusion, Pinus thunbergii extracts can be used as a cosmeceutical agent to regulate sebum production and to alleviate the sebum-involved skin diseases, such as acne and seborrheic dermatitis.

• Applied Chemistry for Engineering
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• v.21 no.6
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• pp.689-693
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• 2010

When natural rubber dissolved in toluene comes into use as a binder of carbon powder, the volatilization of solvent just after the construction of biosensor brought the mechanical robustness on the paste. This characteristic satisfied the pre-requisite condition for the practical use of carbon paste electrode and a biosensor for the determination of hydrogen peroxide was designed. In order to evaluate its electrochemical qualitative and quantitative behaviors, various electrochemical kinetic parameters of the electrode, e.g. the symmetry factor (${\alpha}$, 0.37), the exchange current density ($i_0$, $0.075mAcm^{-2}$), the capacitance of double layer ($C_d$, $9.7{\times}10^{-3}F$), the time constant (${\tau}_A$, 0.92 s), the maximum current ($i_{max}$, $5.92{\times}10^{-7}Acm^{-2}$), the Michaelis constant ($K_M$, $1.99{\times}10^{-3}M$) and others were investigated. Results show that natural rubber is a promising binder of carbon powder.

• Bulletin of the Korean Chemical Society
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• v.27 no.5
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• pp.657-662
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• 2006

Identification of accessible sites in targeted RNAs is a major limitation to the effectiveness of antisense oligonucleotides. A class of antisense oligodeoxynucleotides, known as the “10-23” DNA enzyme or DNAzyme, which is a small catalytic DNA, has been shown to efficiently cleave target RNA at purine-pyrimidine junctions in vitro. We have designed a strategy to identify accessible cleavage sites in the target RNA, which is hepatitis C virus nonstructural gene 3 (HCV NS3) RNA that encodes viral helicase and protease, from a pool of random DNAzyme library. A pool of DNAzymes of 58 nucleotides-length that possess randomized annealing arms, catalytic core sequence, and fixed 5'/3'-end flanking sequences was designed and screened for their ability to cleave the target RNA. The screening procedure, which includes binding of DNAzyme pool to the target RNA under inactive condition, selection and amplification of active DNAzymes, incubation of the selected DNAzymes with the target RNA, and target site identification on sequencing gels, identified 16 potential cleavage sites in the target RNA. Corresponding DNAzymes were constructed for the selected target sites and were tested for RNA-cleavage in terms of kinetics and accessibility. These selected DNAzymes were effective in cleaving the target RNA in the presence of $Mg^{2+}$. This strategy can be applicable to identify accessible sites in any target RNA for antisense oligonucleotides-based gene inactivation methods.

• KSBB Journal
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• v.22 no.1
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• pp.62-65
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• 2007

New strain development strategy using kinetic models and metabolic control analysis was investigated. In this study, previously reported mathematical models describing the enzyme kinetics of intracellular threonine synthesis were modified for mutant threonine producer Escherichia coli TF5015. Using the modified models, metabolic control analysis was carried out to identify the rate limiting step by evaluating the flux control coefficient on the overall threonine synthesis flux exerted by individual enzymatic reactions. The result suggested the production of threonine could be enhanced most efficiently by increasing aspartate semialdehyde dehydrogenase (asd) activity of this strain. Amplification of asd gene in recombinant strain TF5015 (pCL-$P_{aroF}$-asd) increased the threonine production up to 23%, which is much higher than 14% obtained by amplifying aspartate kinse (thrA), other gene in threonine biosynthesis pathway.

• Korean Journal of Food Science and Technology
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• v.14 no.2
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• pp.125-129
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• 1982

Four peroxidase isozymes from horseradish roots (isozymes A, B, C and D) were isolated by chromatography and were thermally inactivated at $70{\sim}97^{\circ}C$ and pH 7.0. The four isozymes had different inactivation rates and the inactivation of each isozymes did not follow first order kinetics. D values of isozymes A, B, C, D and crude enzyme were 594s, 1850s, 2050s, 78s, 130s and z values were $24.0^{\circ}C$, $12.5^{\circ}C$, $18.0^{\circ}C$, $23.7^{\circ}C$ and $24.0^{\circ}C$, respectively. Sephadex gel chromatogram of the thermally treated isozyme C indicated that the shape and molecular weight of the native isozyme changed during inactivation.

• YAKHAK HOEJI
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• v.38 no.2
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• pp.202-210
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• 1994

To investigate the feasibility of mucosal delivery of $[D-Ala^6]$ LHRH, a potent analogue of LHRH, enzymatic proteolysis of $[D-Ala^6]$ LHRH and inhibitory effect of medium chain fatty acid salts(MFA) were studied using rabbit mucosal homogenate. $[D-Ala^6]$ LHRH incubated in homogenates of rectal(RE), nasal(NA) and vaginal(VA) mucosa were assayed by HPLC. The degradation of $[D-Ala^6]$ LHRH followed the first order kinetics. The degradation products were found as $[D-Ala^6]$ $LHRH^{1-7}$(m-i), to a lesser extent, $[D-Ala^6]$ $LHRH^{1-9}$(m-ii) and $[D-Ala^6]$ $LHRH^{1-3}$(m-iii) by the method of amino acid analysis(PITC method). The formation of$[D-Ala^6]$ $LHRH^{1-7}$ was not inhibited by the addition of disodium ethylenediaminetetraacetic acid but inhibited by sodium tauro-24,25-dihydrofusidate, suggesting that endopeptidase 24.11(EP 24.11) cleaves the $Leu^7-Arg^8$ bond of $[D-Ala^6]$ LHRH and is the primary $[D-Ala^6]$ LHRH degrading enzyme. The patterns of $[D-Ala^6]$ LHRH degradation indicated that EP 24.11 exists in each mucosal homogenate with the order of RE>NA>VA. MFA significantly inhibited the proteolysis of $[D-Ala^6]$ LHRH. The addition of sodium caprate(1.0%) or sodium laurate(0.5%) to the each mucosal homogenate completely protected $[D-Ala^6]$ LHRH from the degradation.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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• pp.573-583
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• 2015

B-32 is one of a panel of monoclonal anti-idiotypic antibodies to growth hormone (GH) that we developed. To characterize and identify its potential role as a novel growth hormone receptor (GHR) agonist, we determined that B-32 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assay assays. The results of fluorescence-activated cell sorting, indirect immunofluorescence and competitive receptor binding assays demonstrated that B-32 specifically binds to the GHR expressed on target cells. Next, we examined the resulting signal transduction pathways triggered by this antibody in primary porcine hepatocytes. We found that B-32 can activate the GHR and Janus kinase (2)/signal transducers and activators of transcription (JAK2/STAT5) signalling pathways. The phosphorylation kinetics of JAK2/STAT5 induced by either GH or B-32 were analysed in dose-response and time course experiments. In addition, B32 could also stimulate porcine hepatocytes to secrete insulin-like growth factors-1. Our work indicates that a monoclonal anti-idiotypic antibody to GH (B-32) can serve as a GHR agonist or GH mimic and has application potential in domestic animal (pig) production.

• Advances in Traditional Medicine
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• v.7 no.5
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• pp.477-484
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• 2008

The present study was aimed at investigating the hypouricemic and xanthine oxidase inhibitory activities of the various fractions of the hydromethanolic extract of the leaves of Coccinia grandis L. Voigt (Cucurbitaceae). The leaves of this species was used in traditional medicinal system for the treatment of gout, rheumatism, jaundice, bronchitis, fever, skin eruptions, wounds, etc. The degree of xanthine oxidase inhibition was determined in vitro by measuring the increase in absorbance at 295 nm associated with uric acid formation. Among the fractions tested, the chloroform fraction exhibited highest potency ($IC_{50}$ $17.8\;{\mu}g/ml$). This was followed by the pet-ether ($IC_{50}$ $29.7\;{\mu}g/ml$), ethyl acetate ($IC_{50}$ $41.2\;{\mu}g/ml$) and residual ($IC_{50}$ $47\;{\mu}g/ml$) fractions. The $IC_{50}$ value of allopurinol was $6.1\;{\mu}g/ml$. In addition, the hypouricemic and hepatic xanthine oxidase (XO)/xanthine dehydrogenase (XDH) inhibitory activities of the fractions were examined in vivo using oxonate (280 mg/kg, i.p.) induced hyperuricemic mice. At a dose of 200 mg/kg orally for 7 days, the pet-ether, chloroform and ethyl acetate fractions produced a significant (P < 0.01) reduction in serum urate level and also inhibited hepatic XO/XDH activities when compared to hyperuricemic mice. These inhibitory effects were weaker than that observed for the standard drug, allopurinol (10 mg/kg, p.o.). Lineweaver-Burk analysis of the enzyme kinetics indicated that the mode of inhibition was of a mixed type. These results suggest that the use of Coccinia grandis leaves for the treatment of gout could be attributed to its XO inhibitory activity.