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High Dose of FGF-2 Induced Growth Retardation via ERK1/2 De-phosphorylation in Bone Marrow-derived Mesenchymal Stem Cells

  • Shim, Kwang Yong;Saima, Fatema Tuj;Eom, Young Woo
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.49-56
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    • 2017
  • Fibroblast growth factor (FGF)-2 is one of the most effective growth factors to increase the growth rate of mesenchymal stem cells (MSCs). Previously, we reported that low dose of FGF-2 (1 ng/ml) induced proliferation of bone marrow-derived mesenchymal stem cells (BMSCs) through AKT and ERK activation resulting in reduction of autophagy and senescence, but not at a high dose. In this study, we investigated the effects of high dose FGF-2 (10 ng/ml) on proliferation, autophagy and senescence of BMSCs for long term cultures (i.e., 2 months). FGF-2 increased the growth rate of BMSCs in a dose dependent manner for a short term (3 days), while during long term cultures (2 months), population doubling time was increased and accumulated cell number was lower than control in BMSCs when cultured with 10 ng/ml of FGF-2. 10 ng/ml of FGF-2 induced immediate de-phosphorylation of ERK1/2, expression of LC3-II, and increase of senescence associated ${\beta}$-galactosidase (SA-${\beta}$-Gal, senescence marker) expression. In conclusion, we showed that 10 ng/ml of FGF-2 was inadequate for ex vivo expansion of BMSCs because 10 ng/ml of FGF-2 induced growth retardation via ERK1/2 de-phosphorylation and induction of autophagy and senescence in BMSCs.

Drug Interaction of Cimetidine and Cyclosporine in Human (정상지원자에서 Cimetidine과 Cyclosporine의 약물상호작용)

  • Choi, In;Choi, Jun Shik
    • Korean Journal of Clinical Pharmacy
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    • v.7 no.2
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    • pp.51-63
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    • 1997
  • The effect of cimetidine administration on the pharmacokinetic parameters of cyclosporine were determined in healthy voluteers. This study was performed in 10 volunteers of age ranged 22-48 years and body weight 48-62 kg. This study was performed with cross-over design. Mono cyclosporine and cyclosporine metabolites was extracted from whole blood analysed by fluororescence polarization immune assay (TDX-FLX, Abbott). After coadministration of cimetidine (300 mg) with cyclosporine (300 mg) orally, maximum concentration of mono cyclosporine was significantly increased $1221{\pm}143\;ng/ml\;to\;1562{\pm}184\;ng/ml$ (P<0.05), area under the time curve of cyclosporine (12 hr) also was significantly increased $7478{\pm}829\;ng/ml{\cdot}hr\;to\;9721{\pm}879\;ng/ml{\cdot}hr$ (P<0.05) and absolute baioavailability of cyclosporine was increased $50\pm5.6\%\;to\;57.6\pm6.1\%\;(P<0.05)$ compared to control group. The blood concentrations of cyclopsorine metabolites were significantly decrased (P<0.05) after coadministration of cimetidine. In cimetidine pretreated group, blood mono cyclosporine concentrations were increased significan시y $1220.0\pm203.00\;ng/ml\;to\;1510.0\pm204.00\;ng/ml$ compared with control group (P<0.05). In the mono cyclosporine pharmacokinetic parameter after oral administration absorption rate and maximum concentration were significantly higher in cimetidine coadministered and pretreated group than control group (P<0.05). The ratio of metabolites and mono cyclosporine concentrations was decreased significantly from $70.8\%\;in\;control\;to\;34.8\%$ in coadministration of cimetidine orally. As matter of facts these reults are considered to inhibition of cyclosporine hepatic metabolism and increasing of cyclosporine absorption rate in gastrointestinal tract because of maintaining cyclosporine stability in elevated gastric pH by cimetidine. We considered, it appeares that cimetidine increase bioavailability of cyclosporine by increasing oral absorption and by decreasing hepatic clearance. But the absorption and clearance of cyclosporine was highly variable individually, and therefore we consider that cyclosporine blood level monitoring would be essential in patients with cimetidine co-administration.

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Clinical Evaluation of Pleural Biopsy in the Intrathoracic Lesion with Pleural Effusion (흉막 생검법에 대한 임상적 고찰)

  • 안광수
    • Journal of Chest Surgery
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    • v.26 no.4
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    • pp.298-302
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    • 1993
  • The 40 patients who admitted with chief complaints of pleural effusion and were performed closed thoracostomy and pleural biopsy at the same time with only one incision during the period from Mar,1990. To Feb. 1992. At the department of Thoracic & Cardiovascular Surgery; HanYang University were reviewed retrospectively and the results are as follows: 1. The age of patients ranged from 16 to 73-years old [Mean 44.3-years old]. The peak incidence was fifth decade [25 %] and the next was third decades [22.5 %]. 2. 28 patients were male and 12 patients were female with male preponderance[More than 2 times]. 3. The etiologic of pleural effusion were 25 cases of pulmonary tuberculosis[62.5 %], 8 cases of empyema [20 %], and 7 cases of malignant diseases [17.5 %]. 4. The most common chief complaints were dyspnea[21 cases:29.2%], chest discomfort[16 cases:22.2%], and the coughing with sputum [12 cases: 16.7 %]. 5. The duration of symptom were varied from 3 days to lyear [Mean 3.2 weeks]. 6. The amounts of drained pleural effusion after closed thoracostomy were ranged from 100ml to 2,400 ml [Mean 650 ml], but the amounts in case of malignant pleural effusion were varied from 400ml to 1,700ml [Mean 950ml]. 7. The diagnostic rate was 84.6 % with routine examination of tuberculous pleural effusion [Lymphocyte predominance] and the same rate was acquired by pleural biopsy. 8. The diagnostic rate by pleural biopsy in case of malignant pleural effusion was 57.1% and lower than tuberculous pleural effusion. 9. The etiology of malignant pleural effusion were squamous cell carcinoma [3 cases:42.8 %], adenocarcinoma [2 cases:28.6 %] and metasiatic breast cancer [1 case:14.3%].

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Emodin from Polygonum cuspidatum showed Angiogenesis Inhibiting Activity in vitro (호장근으로부터 분리된 emodin의 혈관신생 억제 활성)

  • Lee, Tae-Kyoo;Kim, Jong-Hwa;So, June-No
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.50-54
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    • 2003
  • Polygonum cuspidatum has been used as a fork medicine for a long time. Emodin was purified from the root of P. cuspidatum by thin layer chromatography (TLC) and preparative high perfomance liquid chromatography (HPLC). The effects of emodin on the migration of endothelial cells and in vitro angiogenesis stimulated with vascular endothelial cell growth factor (VEGF) were examined, using human umbilical vein endothelial cells (HUVECs) and porcine pulmonary arterial endothelial cells (PPAECs). Emodin potently inhibited the VEGF-induced migration of (HUVECs) at relatively low cocentrations $(0.1-10\;{\mu}g/ml)$; the inhibition of endothelial cells by emodin was 75.4% at $0.1\;{\mu}g/ml$ and about 90% at $1\;{\mu}g/ml$. Emodin also inhibited VEGF-induced sprout formation in vitro at concentrations of $0.1-10\;{\mu}g/ml$. Emodin was also evaluated for the inhibitory potential on in vivo angiogenesis in a growing chick embryo chorioallantoic membranes (CAM). At a concentration of $1.0\;{\mu}g/ml$ Per disc, emodin was able to induce avacular zone in the CAMs. These findings suggest that emodin is a potent angiogenesis inhibitor and P. cuspidatum is a useful herb in the development of therapeutics for angiogenesis dependent diseases.

Production of L-sorbose from the resting cells of Acetobacter suboxydans (Acetobacter suboxydans의 휴지 균체에 의한 L-sorbose의 생산)

  • Cho, Won-Dai;Ma, Sang-Jo
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.481-487
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    • 1993
  • The production of sorbose from sorbitol in resting cell system of Acetobacter suboxydans was studied. The conversion of sorbose from sorbitol was markedly influenced by several factors such as the substrate concentration, reaction time, temperature, pH, metal ions, growth factors and aeration in the resting cells. Sorbose production rapidly increased in the range of 6 mg/ml cells with the concentration of 5% sorbitol. For production of sorbose from sorbitol, optimal temperature and pH were $30^{\circ}C$ and 6.0. The production of sorbose from sorbitol was activated by 1 mM of $Al^{+3}$ while inhibited by $Ni^{+2}$. The conversion of sorbitol to sorbose was stimulated by the adding of 1 mM p-aminobenzoic acid and nicotinic acid, respectively. During incubation of 1.5 ml of reaction mixture in 50 ml of Erlenmeyer flask, 5% sorbitol was completly converted to sorbose after 20 hours.

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Development of a One-Step PCR Assay with Nine Primer Pairs for the Detection of Five Diarrheagenic Escherichia coli Types

  • Oh, Kyung-Hwan;Kim, Soo-Bok;Park, Mi-Sun;Cho, Seung-Hak
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.862-868
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    • 2014
  • Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MP-PCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: $5{\times}10^1CFU/ml$ for EHEC, $5{\times}10^3CFU/ml$ for ETEC expressing lt and sth, $5{\times}10^4CFU/ml$ for ETEC expressing stp, $5{\times}10^2CFU/ml$ for EPEC, $5{\times}10^4CFU/ml$ for EAEC, and $5{\times}10^2CFU/ml$ for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.

Production of Kids from In vitro Fertilized Goat Embryos and Their Parentage Assessment Using Microsatellite Markers

  • Malakar, D.;Das, S.K.;Mukesh, M.;Sodhi, M.;Goswami, S.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.842-849
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    • 2007
  • The purpose of the present study was to produce live offspring from in vitro fertilized goat embryos. Oocytes were collected from abattoir ovaries and kept in oocyte collection medium. Oocytes were washed 4-5 times with maturation medium containing medium-199 with 5 ${\mu}g/ml$ FSH, 100 ${\mu}g/ml$ LH, 1 ${\mu}g/ml$ estradiol-$17{\beta}$ 50 ${\mu}g/ml$ gentamycin, 10% inactivated estrus goat serum, and 3% BSA (fatty acid free). Oocytes were placed in 100 ${\mu}l$ drops of maturation medium containing granulosa cell monolayer and incubated in a 5% $CO_2$ incubator at $38.5^{\circ}C$ for 27 h. For capacitation of spermatozoa fresh semen was processed and mixed in 3 ml fertilization TALP medium containing 50 ${\mu}g/ml$ heparin and kept in the above incubator for 2 h. The capacitated spermatozoa were coincubated with matured oocytes for fertilization. Cleaved embryos were separated and cultured in embryo development medium with oviductal cells and 494 embryos were produced. Recipient goats were synchronized with two injections of 15 mg $PGF_{{2}{\alpha}}$/goat 10 days apart. Eighty early stage embryos were transferred into the uterotubal junction of 14 surrogate mothers using laparoscopy techniques. One recipient delivered twin kids, whereas another two recipients each.delivered a single kid The parentage of these kids was evaluated using highly polymorphic co-dominant microsatellites markers. From the present study, it was concluded that live goat kids can be produced from in vitro matured and fertilized goat embryos, to the best of our knowledge for the first time in India.

Clinical Experience with using a Rapid Infuse at an Urban Emergency Department (일개 응급센터에서 급속가압수액주입기의 사용경험)

  • Hong, Chong Kun;Song, Hyoung Gon
    • Journal of Trauma and Injury
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    • v.19 no.1
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    • pp.41-46
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    • 2006
  • Purpose: Hypovolemia is not uncommon among trauma patients in the emergency department (ED). Successful resuscitation of a hypovolemic patient often requires rapid intravenous administration of massive amounts of fluid. A rapid fluid infuser is used in the ED for this purpose, there have been no studies of their clinical uses and effectiveness. We studied clinical experience with a rapid fluid infuser at an urban university hospital in Seoul, Korea. Methods: We reviewed the medical records of 38 patients admitted to the ED with a history of application of a rapid fluid infuser from January 2004 to July 2005. Adult trauma patients older than 15 years of age were included in this study. Clinical data on the patients and the volume of fluid used to achieve a stable blood pressure were extracted from their medical records. Results: The total number of adult trauma patients with a history of application of a rapid fluid infuser from January 2004 to July 2005 in the ED was 16. The mean systolic blood pressure for deciding to apply the rapid fluid infuser was $74.9{\pm}12.7mmHg$. The mean time and volume used to achieve a stable blood pressure were 40.4 min and$2947.3{\pm}1339.2ml$, respectively. In all patients, the amount of fluid infused before using the rapid fluid infuser was between 500 ml and 10,000 ml, compared to 1,000 ml and 6,200 ml with the rapid fluid infuser. The mean amount of fluid per min. via the rapid fluid infuser was 85.5 ml. Vital signs were stabilized in 11 patients, 6 of the 11 were discharged alive. Conclusion: The mean amount of fluid delivered per min. via the rapid fluid infuser was much less than expected; thus, there should be clinical guidelines on volume resuscitation with a rapid fluid infuser in the ED. In the future, prospective, multicenter, clinical-data collection is needed for a more sophisticated study.

THE EFFECT OF HYALURONIC ACID ON MOUSE CALVARIA PRE-OSTEOBLASTS OSTEOGENESIS IN VITRO (히알루론산이 골 형성에 미치는 영향에 관한 실험적 연구)

  • Cho, Yong-Min;Min, Seung-Ki;Kim, Soo-Nam;You, Yong-Ouk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.3
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    • pp.216-225
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    • 2002
  • Hyaluronic acid (HA) is an almost essential component of extracellular matrices. Early in embryogenesis mesenchymal cells migrate, proliferate and differentiate, in part, because of the influence of HA. Since the features of embryogenesis are revisited during wound repair, including bone fracture repair, this study was initiated to evaluate whether HA has an effect on calcification and bone formation in an in vitro system of osteogenesis. Mouse calvaria Pre-osteoblast (MC3T3-E1) cells were cultured in ${\alpha}-MEM$ medium with microorganism-derivative hyaluronic acid that was produced by Strep. zooepidemicus which of molecular weight was 3 million units. The dosages were categorized in each 0.5, 1.0 and 2.0 mg/ml concentration experimental groups. After 2 and 4 days cultures in expeirmental and control groups, the tendency of cell proliferation, MTT assay, protein synthesis ability, collagen synthesis and alkaline phosphatase activity were analysed and bone nodule formation capacity were measured with Alizarin Red S stain after 29 days cultures. The cell proliferation was increased in time, especially the group of 0.5 and 1.0 mg/ml concentration of HA were showed prominent cell proliferation. After 2 and 4 days culture, experimental groups in general were greater cell activity in MTT assay. The protein synthesis was increased in all experimental groups compared to control group, especially most prominent in 1.0 mg/ml concentration group. The collagen synthesis capacity were increased in HA experimental groups, especially prominent in 1.0 mg/ml group and the activity of alkaline phosphatase were increased, especially also prominent in 1.0 mg/ml group, compared to control group. Above these, the activity of mouse carvarial pre-osteoblast cells was showed greater bone osteogenesis activity in all applied HA experimental group, especially group of 1.0 mg/ml concentration of HA.

Determination of Cadmium Ions by Designing an Optode Based on Immobilization of Dithizone on a Triacetylecelluose Membrane in Polluted Soil and Water Samples

  • Tavallali, Hossein;Kazempourfard, Fatemeh
    • Journal of the Korean Chemical Society
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    • v.53 no.2
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    • pp.144-151
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    • 2009
  • An optode for cadmium ion determination has been designed by immobilization of dithizone on triacetylcellose membrane. When the optode membrane is introduced into a real samples containing cadmium, there is a color change from green to red, making it possible to use the change in absorbance at 611 nm as the analytical signal. The sensor could be used in the range of 0.3-3 ${\mu}g\;ml^{-1}$ (2.67-26.67 ${\mu}M$) of $Cd^{2+}$ ions with a limit of detection of 0.025 ${\mu}g\;ml^{-1}$ (25 ng $ml^{-1}$). The response time of optode is within 15 min depending on the concentration of $Cd^{2+}$ ions. It can be easily and completely regenerated by dilute EDTA solution. The effect of different possible interfering species has been examined and was shown the optode has a good selectivity. The results obtained for the determination of cadmium ion in polluted soil and water samples using the proposed optode was found to be comparable with the well-established atomic absorption method.