• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.313-320
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• 2014

The study was conducted to investigate the role of vitamin E in the high altitude hypoxia-induced damage to the intestinal barrier in rats. Sprague-Dawley rats were divided into control (Control), high altitude hypoxia (HH), and high altitude hypoxia + vitamin E (250 mg/kg $BW^*d$) (HV) groups. After the third day, the HH and HV groups were placed in a hypobaric chamber at a stimulated elevation of 7000 m for 5 days. The rats in the HV group were given vitamin E by gavage daily for 8 days. The other rats were given equal volume saline. The results showed that high altitude hypoxia caused the enlargement of heart, liver, lung and kidney, and intestinal villi damage. Supplementation with vitamin E significantly alleviated hypoxia-caused damage to the main organs including intestine, increased the serum superoxide dismutase (SOD) (p< 0.05), diamino oxidase (DAO) (p< 0.01) levels, and decreased the serum levels of interleukin-2 (IL-2) (p< 0.01), interleukin-4 (IL-4) (p<0.001), interferon-gamma ($IFN-{\gamma}$) (p<0.01) and malondialdehyde (MDA) (p<0.001), and decreased the serum erythropoietin (EPO) activity (p<0.05). Administration of vitamin E significantly increased the S-IgA (p<0.001) in ileum and significantly improved the expression levels of occludin and $I{\kappa}B{\alpha}$, and decreased the expression levels of hypoxia-inducible factor 1 alpha and 2 alpha ($HIF-1{\alpha}$ and $HIF-2{\alpha}$), Toll-like receptors (TLR4), P-$I{\kappa}B{\alpha}$ and nuclear factor-${\kappa}B$ p65(NF-${\kappa}B$ P65) in ileum compared to the HH group. This study suggested that vitamin E protectis from intestinal injury caused by high altitude hypoxia environment. These effects may be related to the HIF and TLR4/NF-${\kappa}B$ signaling pathway.

• Journal of Periodontal and Implant Science
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• v.38 no.2
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• pp.153-162
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• 2008

Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study. Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500 ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight $250{\sim}300\;g$). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals) Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area of the defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks. Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.

• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.133-148
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• 1987

Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1604-1609
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• 2016

An HPLC analysis method was developed for standard determination of marmesin as a functional health material in Broussonetia kazinoki extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analyte was detected at 330 nm. The HPLC method was validated in accordance with International Conference on Harmonization guidelines for analytical procedures with respect to specificity, precision, accuracy, and linearity. The limit of detection and quantitation were 6.2 and $18.6{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$>0.9999), and the precision of analysis was satisfactory (less than 0.3%). Recoveries of quantified compound ranged from 100.35 to 101.18%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in B. kazinoki extracts.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.584-592
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• 2004

A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.1-7
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• 2002

Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

• Journal of Chest Surgery
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• v.31 no.5
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• pp.531-535
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• 1998

In 1964, Abbott and Colleagues published the world's first heterotopic heart transplantation technique in the rat. Their method established circulation by end-to-end anastomoses of the graft's aorta and pulmonary artery to the recipient's abdominal aorta and Inferior Vena Cava(IVC), respectively. In 1966, Tomita et al altered Abbott's technique by employing end-to-side rather than end-to-end anastomoses, thus eliminating the hind leg paralysis that sometimes resulted from Abbott's technique. In order to prevent postsuture hemorrhage (since 7-0 silk suture was the finest available at that time), Tomita's aortic anastomosis was done with double up-and-down continuous suture technique. A single layer continuous anstomosis effected the pulmonary artery-IVC anastomosis. The availability of Nylon monofilament suture made it possible for Ono and Lindsey to use a single layer suture technique for the aortic end-to-side anastomosis in their modified rat heart transplantation. We observed survival time between control group and Immunosuppression(Cyclosporine administration, 10mg/Kg${\times}$4 times postoperatively) group after heterotopic heart transplantation in the rat model. The cyclosporine adminstration group survived longer than the control group, thus we concluded that cyclosporine was based on Immunosuppressive drugs.

• Biomolecules & Therapeutics
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• v.6 no.2
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• pp.219-224
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• 1998

The bioequivalence of two clarithromvcin products was evaluated with 16 normal male volunteers (age 23-28 yr, body weight 57.5-75.517g) following single oral dose. Test product was ReYon Clarithromycin tablets (ReYon Pharm. Corp., Korea) and reference product was Klarici $d_{R}$ tablets (Abbott Korea). Both products contain 250 mg of clarithromucin. One tablet of the test or the reference product was administered to the volunteers, respectively, by randomized two period cross-over study (2$\times$2 Latin square method). The determination of clarithromycin was accomplished using a modified agar well diffusion bioassay. As a result of the assay validation, the quantification of clarithromycin in human serum by this technique was possible down to 0.03$\mu$g/ml using 100$\mu$l of serum. The coefficient of variation (C.V.) was less than 10%. Average drug concentrations at each sampling time and pharmacokinetic parameters calculated were not significantly different between two products P>0.05); the area under the curve to last sampling time (24 hr) (AU $Co_{24hr}$ (8.10$\pm$ 1.26 vs 8.22$\pm$ 1.627g . hr/ml), AUC from time zero to infinite (AU $Co_{\infty}$) (8.61 $\pm$ 1.28 vs 8.84$\pm$ 1.71 $\mu$g . hr/ml), maximum plasma concentration ( $C_{msx}$) (0.87$\pm$0.22 vs 0.88$\pm$0.19 $\mu$g/ml) and time to maximum plasma concentration ( $T_{max}$) (2.69 $\pm$0.48 vs 2.56$\pm$ 0.51 hr). The differences of mean AU $Co_{24h}$, $C_{msx}$ and $T_{msx}$ between the two products (1.44, 1.39, and 4.65%, respectively) were less than 20%. The power (1-$\beta$) and treatment difference ($\Delta$) for AU $Co_{24hr}$, and $C_{max}$ were more than 0.8 and less than 0.2, respectivly. Although the power for $T_{max}$ was under 0.8, $T_{max}$. of the two products was not significantly different each other (p>0.05). These results suggest that the bioavailability of ReYon Clarithromycin tablets is not significantly different from that of Klarici $d_{R}$ tablets. Therefore, two products are bioequivalent based on the current results. results.sults.sults.s.s.s.s.s.s.s.

• Korean Journal of Acupuncture
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• v.26 no.4
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• pp.195-209
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• 2009

Objectives : Transient inflammation has been demonstrated to alter visceral sensory function in animal models and acute mucosal inflammation may precede the manifestation of visceral hyperalgesia. Thus in this study we compared effects of herbal acupuncture of Nidus Vespae (NV) applied to the different acupoints in the acute colitis induced by trinitrobenzenesulphonic acid (TNBS) intracolonic injection in rats. Methods : In Male Sprague-Dawley rats, weighing 250 ~ 400 g, TNBS (5 mg/kg) was infused intrarectally through a silicon rubber catheter into the anus under isoflurane anaesthesia. Under general anesthesia, acupoints of LI4 (Hapkok), SI25 (Cheonchu), ST36 (Joksamni), BL25 (Daejangsu) were intramuscularly injected by NV. Expressions of cFos protein in the periaqueductal gray (PAG), locus coeruleus (LC), nucleus of solitary tract (Sol), and the 6th lumbar spinal cord (L6 s.c.) were observed at 24 hrs after TNBS induced colitis by immunohistochemistry. Results : The expression of c-Fos protein in L6 s.c., Sol, LC and PAG increased 24 hrs after TNBS injection into colorectum as compared to normal group. NV herbal acupuncture also inhibited the expression of c-Fos protein in Sol but not L6 s.c., LC, and PAG. NV to ST36 inhibited significantly the c-Fos expression in Sol and PAG. NV to ST25 inhibited the c-Fos protein expression all over the observation area. NV to BL25 showed the inhibitory effects in the areas except LC. Whether or not a role of endogenous opioids, intrathecal injection of naltrexone (30 ug / 30 ul) was applied before the 2nd herbal acupuncture treatment 24 hrs after TNBS-induced colitis in rat. Naltrexone reversed the inhibition of c-Fos protein expression in the spinal cord and brainstem under different conditions such as type of herbal acupuncture compound and choice of acupoint. Conclusions : In summary, these data show that herbal acupuncture of NV inhibits signal pathways such as spinal cord and brain stem ascending hypersensitivity of colorectum after TNBS induced colitis. This effect may be mediated by acupoints through the endogenous opioid system involving the pain modulation.

• Journal of Yeungnam Medical Science
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• v.18 no.1
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• pp.51-58
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• 2001

Background: Cholestatic hepatitis is failure of bile to reach the duodenum with hepatocellular damage and no demonstable obstruction of the major bile ducts. The prognosis is usually good with recovery in less than 4 weeks after withdrawal of the offending drug. However, a prolonged course lasting over 3 months is possible and, in rare cases, progression to ductopenia with development of a vanishing bile duct syndrome occurs. A differential diagnosis with other causes of Chronic liver disease is needed. Materials and Methods: From January 1991 through January 2000, 14 patients diagnosed as cholestatic hepatitis by liver biopsy were included. The possible causative drug, clinical features, laboratory findings, and progression of cholestatic hepatitis were evaluated. The semiquantitative study of liver lesions was performed by two independent observers. Results: Causes of cholestatic hepatitis are 5 cases of oriental medicine, 3 cases of anti-tuberculosis medication, 1 case of ticlopidine and antibiotics and 4 cases of unknown causes. The clinical features of cholestatic hepatitis were jaundice, itching, urine color change, and general weakness. During 6 to 30 months, LFT of 5 patients showed prolonged elevation. Elevated total cholesterol ${\geq}$250 mg/dL in 6 patients, pheripheral blood eosinophilia in 5 patients, auto-antibody positive in 6 patients were observed respectively. The biopsies showed intralobular bilirubinostasis with a mixed portal inflammatory infiltration. Conclusion: In cholestatic hepatitis, durations of abnormal LFT are variable regardless of causative drugs. If cholestatic hepatitis progresses toward chronic course, viral hepatitis, primary biliary cirrhosis, and autoimmune hepatitis should be differentially diagnosed and sequential liver biopsies are needed.