• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.1045-1048
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• 2007

Two genes, dps encoding decaprenyl diphosphate synthase and dxs encoding 1-deoxy-D-xylulose 5-phosphate synthase, were isolated from Rhizobium radiobacter ATCC 4718. DNA sequencing analysis of the dps and dxs genes revealed an open reading frame of 1,077 bp and 1,920 bp, respectively. The heterologous expression in Escherichia coli BL21(DE3) was carried out in order to identify their functions. Recombinant E. coli BL21(DE3) harboring the dps gene produced $CoQ_{10}$ as well as $CoQ_8$ and $CoQ_9$, whereas E. coli harboring only the dxs gene produced more $CoQ_8$ compared with the wild-type E. coli. Additionally, the coexpression of dps and dxs genes in E. coli was carried out. The recombinant E. coli harboring only the dps gene produced $0.21{\pm}0.04\;mg/l$ of $CoQ_{10}$, whereas the coexpressed E. coli with dps and dxs genes produced $0.37{\pm}0.07\;mg/l$ of $CoQ_{10}$. HPLC analysis also showed that the $CoQ_{10}$ fraction (100% of the total CoQs distribution) was increased from $15.86{\pm}0.66%$ (only dps) to $29.78{\pm}1.80%$ (dps and dxs).

• Journal of Pharmaceutical Investigation
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• v.28 no.3
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• pp.193-198
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• 1998

Bioequivalence of two cilostazol tablets, the $Pletaal^{TM}$ (Korea Otsuka Pharmaceutical Co., Ltd.) and the $Losazol^{TM}$ (Kyoung Dong Pharmaceutical Co., Ltd.), was evaluated according to the guideline of KFDA. Fourteen normal male volunteers (age $20{\sim}28$ years old) were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After two tablets containing 50 mg of cilostazol were orally administered, blood was taken at predetermined time intervals and the concentration of cilostazol in plasma was determined with an HPLC method using UV detector. The pharmacokinetic parameters $(C_{max},\;T_{max}\;and\;AUC_t)$ were calculated and ANOVA was utilized for the statistical analysis of parameters. The results showed that the differences in $C_{max},\;T_{max}\;and\;AUC_t$ between two tablets were 3.14%, 10.0% and 7.35%, respectively. The powers $(1-{\beta})$ for $C_{max},\;T_{max}\;and\;AUC_t$ were 89.67%, 80.97% and 83.87%, respectively. Detectable differences $({\Delta})$ and confidence intervals were all less than 20% except $T-{max}$, but confidence interval of $T_{max}$ was also less than 20% at the significance $level({\alpha})$ of 0.1. All of these parameters met the criteria of KFDA for bioequivalence, indicating that $Losazol^{TM}$ tablet is bioequivalent to $Pletaal^{TM} tablet$.

• BMB Reports
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• v.40 no.6
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• pp.1016-1020
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• 2007

In this study, dioscin was isolated from Dioscoreae Rhizoma (DR), which is the rhizome of Dioscorea batatas DECNE. that inhabits broad areas of Korea and Japan. To determine whether dioscin induced growth hormone (GH) release, we evaluated its induction effects on GH release both in vitro and in vivo. The 70% methanol extract of DR, and its n-hexane and n-BuOH fractions, induced rat GH (rGH) release in rat pituitary cells 10-fold, 8-fold, and 5-fold higher than the control ($0.36{\pm}0.02 nM$), respectively (p < 0.05 each). The dioscin-induced rGH release of the cells was concentration-dependent and its $ED_{50}$ was $1.14{\times}10^{-5} M$. Within 90 minutes after intravenous administration of $10{\mu}g$/kg (p < 0.05 at $t_{max}$), dioscin caused the greatest increase in rGH concentration ($C_{max}$) in the rat plasma ($34.16{\pm}14.10 ng/ml$) (n = 4), which was twice as high as the control group ($12.88{\pm}3.29 ng/ml$) (n = 27).

• BMB Reports
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• v.40 no.6
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• pp.979-985
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• 2007

Induction of growth hormone (GH) by Glycyrrhizae Radix (GR), one of the most popular herbal medicine, and its major ingredients were studied in rat pituitary cells in vitro and in vivo assay. The MeOH extract and the n-hexane (HX) fraction of GR induced rat GH (rGH) release up to 1.89 times ($0.34{\pm}0.04 nM$) and 4.59 times ($0.83{\pm}0.03 nM$), compared to the basal level (p < 0.05). Among many ingredients isolated and purified from GR both glycyrrhetinic acid and glycyrrhizin induced significantly rGH release compared to the control (p < 0.05). After an intravenous injection of rat growth hormone releasing hormone (rGHRH) ($10{\mu}g$/kg) as positive control, in SD rats, $T_{max}$ of plasma rGH level was 10 min, $C_{max}$ was $3.84{\pm}0.01 nM$ (n = 3), and enhanced plasma rGH level returned to the baseline in 90 min. Both $AUC_{0-90}$ (area under the curve) of plasma rGH level after HX fraction and that after rGHRH administration were increased significantly from the basal level, respectively (p < 0.01). In conclusions, HX fraction is the most active fraction of MeOH extract of GR in rGH induction.

• Archives of Pharmacal Research
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• v.26 no.11
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• pp.929-936
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• 2003

Methanol extract (MeOH), n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), butanol (BuOH) and aqueous ($H_2O$) fractions of Eucommiae Cortex including geniposidic acid (GA), geniposide (GP) and aucubin (AU) were tested for their therapeutic efficacy on osteoporosis. The contents of GA, GP and AU in the cortex and leaf of Eucommia ulmoides Oliver were quantified by HPLC. The effect of Eucommiae Cortex on the induction of growth hormone (GH) release was studied by using rat pituitary cells. The proliferation of osteoblast-like cells increased by herbal extracts was assayed using a tetrazolium (MTT), alkaline phosphatase (ALP) activity, and [$^3H$]-proline incorporation assays. The inhibition of osteoclast was studied by using the coculture of mouse bone marrow cells and ST-2 cells. As a result, the GA, GP and AU were present in the cortex more than in the leaf of E. ulmoides Oliver. The MeOH (1mg/mL), Hx, $CHCl_3$ and EA fractions (each 20 $\mu$ g/mL) had potent induction of GH release. The $CHCl_3$ exhibited the potent proliferation of osteoblasts. The AU, GP and GA were increased proliferation of osteoblasts. In addition, GA ($IC_{50}: 4.43{\times}10^{-7}$M), AU and GP were significantly inhibited proliferation of osteoclast. In summary, it is thought that the components in a part of the fractions of Eucommiae Cortex participate in each step of mechanism for activating osteoblast to facilitate osteogenesis, and suppress osteoclast activity to inhibit osteolysis.

• Toxicological Research
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• v.24 no.4
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• pp.245-251
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• 2008

Dietary restriction (DR) is the most efficacious intervention for retarding the deleterious effects of aging. DR increases longevity, decreases the occurrence and severity of age-related diseases, and retards the physiological decline associated with aging. The beneficial effects of DR have been mostly studied in non-neuronal tissues. However, several studies have showed that DR attenuate neuronal loss after several different insults including exposure to kainate, ischemia, and MPTP. Moreover, administration of the non-metabolizable glucose analog 2-deoxy-D-glucose (2DG) could mimic the neuroprotective effect of DR in rodent, presumably by limiting glucose availability at the cellular level. Based on the studies of chemically induced DR, it has been proposed that the mechanism whereby DR and 2DG protect neurons is largely mediated by stress response proteins such as HSP70 and GRP78 which are increased in neurons of rats and mice fed a DR regimen. In addition, DR, as mild metabolic stress, could lead to the increased activity in neuronal circuits and thus induce expression of neurotrophic factors. Interestingly, such increased neuronal activities also enhance neurogenesis in the brains of adult rodents. In this review, we focus on what is known regarding molecular mechanisms of the protective role of DR in neurodegenerative diseases and aging process. Also, we propose that DR is a mild cellular stress that stimulates production of neurotrophic factors, which are major regulators of neuronal survival, as well as neurogenesis in adult brain.

• Molecules and Cells
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• v.28 no.1
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• pp.67-71
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• 2009

Estrogen receptor ${\alpha}$ ($ER{\alpha}$) mediates the mitogenic effects of estrogen. $ER{\alpha}$ signaling regulates the normal growth and differentiation of mammary tissue, but uncontrolled $ER{\alpha}$ activation increases the risk to breast cancer. Estrogen binding induces ligand-dependent $ER{\alpha}$ activation, thereby facilitating $ER{\alpha}$ dimerization, promoter binding and coactivator recruitment. $ER{\alpha}$ can also be activated in a ligand-independent manner by many signaling pathways, including protein kinase A (PKA) signaling. However, in several $ER{\alpha}$-positive breast cancer cells, PKA inhibits estrogen-dependent cell growth. FoxH1 represses the transcriptional activities of estrogen receptors and androgen receptors (AR). Interestingly, FoxH1 has been found to inhibit the PKA-induced and ligand-induced activation of AR. In the present study, we examined the effects of PKA activation on the ability of FoxH1 to represses $ER{\alpha}$ transcriptional activity. We found that PKA increases the protein stability of FoxH1, and that FoxH1 inhibits PKA-induced and estradiol-induced activation of an estrogen response element (ERE). Furthermore, in MCF7 cells, FoxH1 knockdown increased the PKA-induced and estradiol-induced activation of the ERE. These results suggest that PKA can negatively regulate $ER{\alpha}$, at least in part, through FoxH1.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.570-575
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• 1998

To develop a cotton derivatives with prolonged antimicrobial activities, homosulfamine (Hs) was coupled to cotton xanthate (CX) via chelate bond in the presence of Cu(II) ion by one- and two-bath processes. In one-bath process, CX was treated with Cu(II)-Hs solution. In two-bath process, CX was treated with Cu(II) ion solution to produce CX-Cu(II) complex, which was isolated and treated in turn with Hs solution. Effects of concentration, Cu(II)/Hs ratio, and pH on the binding of Hs were investigated at $10{\circ}C$ . In one-bath process, binding of hs took place readily with optimum pH around 5-6. the amount of binding increased to give a maximum within 5 min and decreased slowly to establish an equilibrium withhin an hour. In two-bath process, binding of Hs was much lower than that of one-bath process. Release of Hs from CX-Cu(II)-Hs was investigated by batch and flow method. antimicrobial activiteis of CX-Cu(II)-Hs was investigated by batch and flow method. Antimicrobial activities of CX-Cu(II)-Hs were tested against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli and it showed prolonged activity compared to that of free Hs.

• Journal of Pharmaceutical Investigation
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• v.28 no.4
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• pp.289-294
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• 1998

Bioequivalence of two aceclofenac tablets, the $Airtal^{TM}$ (Daewoong Pharmaceutical Co., Ltd.) and the $Senital^{TM}$ (Hana Pharmaceutical Co., Ltd.), was evaluated according to the guideline of KFDA. Fourteen normal male volunteers (age $20{\sim}29$ years old) were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 100 mg of aceclofenac was orally administered, blood was taken at predetermined time intervals and the concentration of aceclofenac in plasma was determined with an HPLC method using UV detector. The pharmacokinetic parameters ($C_{max}$, $T_{max}$ and $AUC_t$) were calculated and ANOVA was utilized for the statistical analysis of parameters. The results showed that the differences in $C_{max}$, $T_{max}$ and $AUC_t$ between two tablets were 3.69%, 2.44% and 0.51%, respectively. The powers $(1-{\beta})$ for $C_{max}$, $T_{max}$ and $AUC_t$ were 87.85%, 98.70% and more than 99%, respectively. Detectable differences $({\Delta})$ and confidence intervals were all less than ${\pm}20%$. All of these parameters met the criteria of KFDA for bioequivalence, indicating that $Senital^{TM}$ tablet is bioequivalent to $Airtal^{TM}$ tablet.

• Archives of Pharmacal Research
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• v.26 no.1
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• pp.34-39
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• 2003

The traditional Asian medicinal herb, roots of Astragalus (A.) membranaceus (Leguminosae), is used for many purposes, some of which are purported to stimulate the release of growth hormone in vivo. Extracts of A. membranaceus were tested to determine whether they stimulate the release of growth hormone in rat pituitary cell culture. A. membranaceus was extracted sequentially with 80% ethanol (fraction A), n-hexane (fraction B); the test compound from the herbal extraction was isolated using silica gel column chromatography and was identified with spectral data. Test compound was also extracted by traditional boiling water methods. Induction of growth hormone in pituitary cell culture was conducted with isolated compounds and extracted fractions of A. Radix (dried roots of A. membranaceus). The fraction A was not active in the rat pituitary cell culture, but the fraction B derived from the ethanol fraction stimulated the release of growth hormone in culture. Six compounds from fraction B (1-6) were isolated and identified previously. The compounds 1,2-benzendicarboxylic acid diisononylester (1), $\beta$-sitosterol (2), and 3-Ο-$\beta$-D-galactopyranosyl-$\beta$-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z, 12Z-octadecadienoic acid (4), stigmast-4-en-6$\beta$-o1-3-one (6) and 98-E, a mixture of 1'-9, 12-octadecadienoic acid (Z,Z)-2',3'-dihydroxy-propylester (7) and 1'-hexadecanoic acid-2',3'-dihydroxy-propylester (8) stimulated the release of growth hormone in the rat pituitary cell culture significantly compared to the control. In conclusions, four compounds isolated from extracts of A. Radix induced growth hormone release in the rat pituitary cell culture. The 98-E isolate was the most active inducer of growth hormone release.