• 제목/요약/키워드: ${\alpha}$-Method

Search Result 3,584, Processing Time 0.025 seconds

High-Level Production of High-Purity Human and Murine Recombinant Prion Proteins Functionally Compatible to In Vitro Seeding Assay

  • Hwang, Hae-Gwang;Kim, Dae-Hwan;Lee, Jeongmin;Mo, Youngwon;Lee, Se-Hoon;Lee, Yongjin;Hyeon, Jae Wook;Lee, Sol Moe;Cheon, Yong-Pil;Choi, Eun-Kyoung;Kim, Su Yeon;Lee, Yeong Seon;Son, Young-Jin;Ryou, Chongsuk
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.10
    • /
    • pp.1749-1759
    • /
    • 2018
  • Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

Production of Transgenic Animals by the Testis-Mediated Gene Transfer II. Production of Transgenic Korean Native Goats (정소실질내 유전자 도입에 의한 형질전환동물의 생산 II. 형질전환 한국재래산양의 생산)

  • 윤창현;장규태;김성현;박미령;주학진;오석두;이병오
    • Korean Journal of Animal Reproduction
    • /
    • v.23 no.1
    • /
    • pp.13-18
    • /
    • 1999
  • The totipotential spermatogonial stem cells of adult testis which give rise to mature sperm cells is well-known to incorporate foreign DNA as well as those of somatic cells. Also, the integration rates of foreign DNA after haploid stages are generally known to decrease and /or is simply bound foreign DNA into the sperm plasma membrane. To overcome these problems, liposome and DNA complexes were used to determine how direct injection of these complexes into testis were integrated into sperm genome and resulted in transgenic offspring. To study this purpose, cation liposome was gently mixed with WAF/hGH DNA (1 : 2) and the complexes were injected into testis. At 10, 20, 40, 60, and 80 days after direct injection into testis, mature sperm cells were recovered by using artificial virgin method from two goats and each semen except a part of semen used for DNA analysis such as PCR or Southern blotting was cryopreserved for the artificial insemination. The results obtained in this study are summarized as follows. 1. By PCR, the presence of exogenous DNA was confirmed up to 80 days after injection with liposome/DNA complexes. The highest integration rates were obtained at day 40 after direct injection. This results suggested that spermatogonial stem cells were integrated exogenous DNA into their genome. 2. Among 23 Korean Native Goats which were artificially inseminated, 4 goats resulted in pregnancy and produced 7 young goats. 3. Two young goats were confirmed as a transgenic by PCR and Southern blot analysis. Therefore, our results suggested that testis-mediated gene transfer can be used as a feasible tools for the production of transgenic livestock.

  • PDF

Effect of Astragalus Membranaceus Extract against Improvement of Myelosuppression and Quality of Life in 5-Fluorouracil Treated Mice (황기 추출물이 5-Fluorouracil을 투여한 생쥐의 골수억제 및 삶의 질에 미치는 영향)

  • Kwon, Chang-Hyun;Yoo, Hwa-Seung;Bang, Sun-Hwi;Lee, Young-Min;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan
    • The Journal of Internal Korean Medicine
    • /
    • v.28 no.2
    • /
    • pp.304-320
    • /
    • 2007
  • Objective : To evaluate the effect of Astragalus Membranaceus Extrac (AME) on myelosuppression, activity and immune modulation in 5-fluorouracil (5-FU) treated mice. Method : We carried out complete blood count, histological analysis of bone marrow, and cell colony forming assay for hematopoietic progenitor to evaluate the effect of AME on myelosuppression and conducted swimming test, survival rate, nitric oxide (NO) assay, 51Cr release assay in natural killer cell, mRNA expression of $IL-1{\beta}$, IL-2, IL-4, IL-6, IL-10, $TNF-{\alpht}$, $IFN-{\gamma}$, $TGF-{\beta}$ and GM-CSF in spleen cells to evaluate the effect of AME on quality of life (QOL). Results : AME improved 5-FU induced myelosuppression and peripheral blood count was recovered effectively, had significant efficacy to protect against chemotherapy induced marrow-destruction and on hematopoiesis compared with the control group, improved increase survival rate and the swimming time, had a stimulatory effect on macrophage activation and NK cell activity, and up-regulated cytokine gene transcription (IL-2, IL-6, $IFN-{\gamma}$) in murine immunologic system. Conclusion : We can conclude that AM is an effective herbal agent for improvement of myelosuppression and QOL in 5-FU treated mice.

  • PDF

AGL gene mutation and clinical features in Korean patients with glycogen storage disease type III (한국인 제3형 당원병 환자의 임상상 및 AGL 유전자형)

  • Ko, Jung Min;Lee, Jung Hyun;Kim, Gu-Hwan;Yoo, Han-Wook
    • Journal of The Korean Society of Inherited Metabolic disease
    • /
    • v.6 no.1
    • /
    • pp.15-23
    • /
    • 2006
  • Purpose: Glycogen storage disease type III (GSD-III), is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), which is responsible for the debranching of the glycogen molecule during catabolism. The disease has been demonstrated to show clinical and biochemical heterogeneity, reflecting the genotype-phenotype heterogeneity among different patients. In this study, we analyzed mutations of the AGL gene in three unrelated Korean GSD-III patients and discussed their clinical and laboratory implications. Methods: We studied three GSD-III patients and the clinical features were characterized. Sequence analysis of 35exons and part exon-intron boundaries of the AGLgene in patients were carried out by direct DNA sequencing method using genomic DNA isolated from patients' peripheral leukocytes. Results: The clinical features included hepatomegaly (in all patients), seizures (in patient 2), growth failure (in patients 1), hyperlipidemia (in patients 1 and 3), raised transaminases and creatinine kinase concentrations (in all patients) and mild EKG abnormalities (in patients 2). Liver transplantation was performed in patient 2due to progressive hepatic fibrosis. Administration of raw-corn-starch could maintain normoglycemia and improve the condition. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 with c.1510_1511insT (p.Y504LfsX10), and patient 3 with c.3416 T>C (p.L1139P) and c.l735+1 G>T (Y538_R578delfsX4) mutations. Except R428K mutation, 4 other mutations identified in3 patients were novel. Conclusion: GSD-III patients have variable phenotypic characteristics resembling GSD-Ia. The molecular defects in the AGL gene of Korean GSD-III patients were genetically heterogeneous.

  • PDF

Meauring Method of Magnetic Particles' Coercivity Distribution and Its Applications (자성분말의 보자력 분포도 측정방법과 그 응용)

  • 홍양기;박상준;정홍식
    • Journal of the Korean Magnetics Society
    • /
    • v.5 no.3
    • /
    • pp.216-221
    • /
    • 1995
  • Both manufacturing parameters and particles' chemical composition controll coercivity and its distribution of magnetic particles. The coercivity and its distribution are important properties for high density magnetic recording, so these are used as tool for evaluation of reproducibility of magnetic particles. We report in this paper the applications of dM/dH versus H curve, which is derived from magnetic hysteresis loop, to the evaluation of coercivity distribution of magnetic particles and oxidation studies of recording metal particles. The coercivity distri-bution can be estimated from the full width half rnaximun (FWHM) and the peak shape of the dM/dH versus H curve. The peak shape of the curve depends upon distribution of particles' coercivity. The peak of dM/dH versus H curve becomes broad and lor is splitted into two or rmre peaks. It depends on uniformity of particles' coercivity. When the coercivity difference between Ba-Ferrite and ${\gamma}-Fe_{2}O_{3}$ is larger than 600 Oe, the peak becomes broad and is consequently splitted into two peaks. Ununiformly substituted Ba-Ferrite particles show broad peak. It is apparent that the analysis of the curve is one of sensitive measuring techniques for determination of coercivity distri-bution and studies on magnetic properties of substituted Ba-Ferrite and oxidation of magnetic recording metal particles.

  • PDF

Volatile Aroma Compounds and Their Characteristics of Labiatae by Solid-Phase Microextraction (SPME) (Solid-Phase Microextraction (SPME)에 의한 꿀풀과 약초의 향기성분과 그 특성)

  • Song, Yong-Eun;Ku, Chang-Sub;Mun, Sung-Phil;Ryu, Ji-Sung;Kim, Dae-Hyang;Choi, Joung-Sik;Choi, Yeong-Geun
    • Korean Journal of Medicinal Crop Science
    • /
    • v.10 no.2
    • /
    • pp.120-125
    • /
    • 2002
  • This study was carried out to find the possibility of use of Solid Phase Microextraction (SPME) for extracting the volatile aroma compounds in the five aromatic plants (Agastache rugosa O. Kuntze, Mentha arvensis Linne, Thymus quinquecostatus Celakovsky, Elsholtzia splendens Nakai, Schizonepta tenuifolia Briquet) belongs to the Labiatae. In the result of the analysis, the volatile aroma compounds were mainly composed monoterpene alcohol (linanol, menthol, ${\alpha}-terpineol$, borneol), monoterpene ketone (limonene, menthone) and sesquiterpene (trans-caryophyllene,${\delta}-cadinene)$. The volatile aroma compounds of Agastache rugosa O. Kuntze and Mentha arvensis Linne were extracted by SPME more identified than the SDE. However, Schizonepta tenuifolia Briquet more identified by the SDE and in Elsholtzia splendens Nakai similar to the SDE. Especially, the SPME showed the sesquiterpene contents was more than the SDE. The major volatile aroma compounds were difference but the composition of those between the SPME and the SDE showed no difference. Within the results, the SPME showed the most convenient and a rapid extraction method to analysis of the volatile aroma compounds.

Synthesis of Fe3O4-δ Using FeC2O4·2H2O by Thermal Decomposition in N2 Atmosphere (N2분위기에서 FeC2O4·2H2O의 열분해에 의한 Fe3O4-δ합성)

  • Park, Won-Shik;Oh, Kyoung-Hwan;An, Suk-Jin;Suhr, Dong-Soo
    • Korean Journal of Materials Research
    • /
    • v.22 no.5
    • /
    • pp.253-258
    • /
    • 2012
  • Activated magnetite ($Fe_3O_{4-{\delta}}$) was applied to reducing $CO_2$ gas emissions to avoid greenhouse effects. Wet and dry methods were developed as a $CO_2$ removal process. One of the typical dry methods is $CO_2$ decomposition using activated magnetite ($Fe_3O_{4-{\delta}}$). Generally, $Fe_3O_{4-{\delta}}$ is manufactured by reduction of $Fe_3O_4$ by $H_2$ gas. This process has an explosion risk. Therefore, a non-explosive process to make $Fe_3O_{4-{\delta}}$ was studied using $FeC_2O_4{\cdot}2H_2O$ and $N_2$. $FeSO_4{\cdot}7H_2O$ and $(NH_4)_2C_2O_4{\cdot}H_2O$ were used as starting materials. So, ${\alpha}-FeC_2O_4{\cdot}2H_2O$ was synthesized by precipitation method. During the calcination process, $FeC_2O_4{\cdot}2H_2O$ was decomposed to $Fe_3O_4$, CO, and $CO_2$. The specific surface area of the activated magnetite varied with the calcination temperature from 15.43 $m^2/g$ to 9.32 $m^2/g$. The densities of $FeC_2O_4{\cdot}2H_2O$ and $Fe_3O_4$ were 2.28 g/$cm^3$ and 5.2 g/$cm^3$, respectively. Also, the $Fe_3O_4$ was reduced to $Fe_3O_{4-{\delta}}$ by CO. From the TGA results in air of the specimen that was calcined at $450^{\circ}C$ for three hours in $N_2$ atmosphere, the ${\delta}$-value of $Fe_3O_{4-{\delta}}$ was estimated. The ${\delta}$-value of $Fe_3O_{4-{\delta}}$ was 0.3170 when the sample was heat treated at $400^{\circ}C$ for 3 hours and 0.6583 when the sample was heat treated at $450^{\circ}C$ for 3 hours. $Fe_3O_{4-{\delta}}$ was oxidized to $Fe_3O_4$ when $Fe_3O_{4-{\delta}}$ was reacted with $CO_2$ because $CO_2$ is decomposed to C and $O_2$.

Bioequivalence of Prepulsid Tablet to Cisaple Tablet (Cisapride 5 mg) (프레팔시드 정(시사프리드 5 mg)에 대한 시사플 정의 생물학적 동등성)

  • Kwak, Son-Hyuk;Nam, Jin-Kyung;Jiang, Ge;Han, Jung-Hee;Woo, Jong-Soo;Rhee, Gye-Ju;Park, Jong-Woo;Koo, Sun-Hoe;Hwang, Sung-Joo
    • Journal of Pharmaceutical Investigation
    • /
    • v.30 no.1
    • /
    • pp.55-59
    • /
    • 2000
  • Bioequivalence of two cisapride tablets, test drug ($Cisple^{\circledR}$ tablet: Hanmi Pharm Co., Ltd.) and reference drug ($Prepulsid^{\circledR}$ tablet: Janssen Pharm. Co., Ltd.), was evaluated according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty two healthy male volunteers were divided randomly into two groups and administered the drug orally at the dose of 10 mg as cisapride in a $2{\times}2$ crossover study. There was a week washout period between administrations. Blood samples were taken at predetermined time intervals for 36 hr and the plasma concentration of cisapride was determined by a HPLC method. $AUC_{0-36hr}$ (area under the plasma concentration-time curve from time zero to 36 hr), $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were estimated from the plasma drug concentration-time data. Analysis of variance (ANOVA) revealed no difference in $AUC_{0-36hr},\;C_{max}\;and\;T_{max}$ between two products. The apparent differences of these parameters between two products were less than 20% (i.e., 5.38, 6.17 and 0.00% for $AUC_{0-36hr},\;C_{max}\;and\;T_{max},$ respectively). The powers $(1-\beta)$ for $AUC_{0-36hr},\;C_{max}\;and\;T_{max}$ were over 0.9. Minimal detectable differences $(\Delta)$ at ${\alpha}=0.05,\;1-{\beta}=0.8$ were less than 20% (i.e. 17.67, 14.84 and 19.72% for $AUC_{0-36hr},\;C_{max}\;and\;T_{max},$ respectively). The 90% confidence intervals $(\delta)$ for these parameters were also within ${\pm}20%$ $(i.e.\;-4.97\;{\le}{\delta}{\le}\;15.73,\;-2.53{\le}{\delta}{\le}\;14.86\;and\;-11.55{\le}{\delta}{\le}\;11.55$ for $AUC_{0-36hr},\;C_{max}\;and\;T_{max},$ respectively). These results satisfied the criteria of KFDA guidelines for bioequivalence, indicating the two tablets of cisapride were bioequivalent.

  • PDF

Anti-bacterial Effects of Aqueous Extract Purified from the Immature Cone of Red Pine (Pinus densiflora) (미성숙 솔방울 열수추출물의 항균성)

  • Jeong, Kyung Hui;Hwang, In Sik;Kim, Ji Eun;Lee, Young Ju;Kwak, Moon Hwa;Lee, Young Hee;Lee, Jae Ho;Hwang, Dae Youn;Jung, Young Jin
    • Textile Coloration and Finishing
    • /
    • v.26 no.1
    • /
    • pp.45-52
    • /
    • 2014
  • Red pine (Pinus densiflora) is widely used traditional medicine, pharmacological and nutritional values from which the phytochemical compounds are derived. The present study was aimed to examine the antibacterial effects in the absence and presence of a immature red pine cone extract against 13 microorganisms. The components in the aqueous extract from immature red pine cone were identified by GC-MS. About 1.4% of total polyphenolic compound was measured in aqueous extract collected from immature red pine cone. Also, the high concentration of ${\beta}$-phellenandrene, ${\alpha}$-pinene, limonene, bornyl acetate and aldehyde was detected in total ion chromatograms. Of total 13 microorganisms, 4 microorganisms including Pseudomonas aeruginosa, Vibrio cholera, Listeria monocytogenes, Klebsiella pneumonia were effectively killed by aqueous extract of immature red pine cone. The highest anti-bacterial effect was detected in P. aeruginosa, followed by V. cholera, L. monocytogenes and K. pneumonia. In case of P. aeruginosa, the largest diameter of inhibition zone was maintained to 1/2 solution treated cells and slightly decreased at 1/4 and 1/8 solution treated cells. Also, in test used V. cholera and L. monocytogenes, the inhibition zone was strongly formed in only 1 and 1/2 solution treated cells, while K. pneumonia showed the very small diameter of inhibition zone in all concentrations. Therefore, these results suggested that the aqueous extracts of immature red pine cone should be considered as a new and potentially important anti-bacterial substrate to effectively prevent the microbial infection and penetration.

Growth and Scintillation Characteristics of CsI(Br) Single Crystals (CsI(Br) 단결정의 육성과 섬광특성)

  • Oh, M.Y.;Jung, Y.J.;Lee, W.G.;Doh, S.H.;Kang, K.J.;Kim, D.S.;Kim, W.;Kang, H.D.
    • Journal of Sensor Science and Technology
    • /
    • v.9 no.5
    • /
    • pp.341-349
    • /
    • 2000
  • CsI(Br) single crystals doped with 1, 3, 5 or 10 mole% $Br^-$ ions, as an activator, were grown by Czochralski method. The lattice structure of grown CsI(Br) single crystal was bcc and its lattice constant was $4.568\;{\AA}$. The absorption edge of the CsI(Br) single crystals was observed at 243 nm. The spectral range of the luminescence excited by 243 nm of wavelength was $300{\sim}600\;nm$, and its peak emission appeared at 440 nm. The luminescence intensity was maximum when CsI(Br) was doped with 3 mole % $Br^-$ ions. The energy resolutions of the CsI(Br) scintillator doped with 3 mole % $Br^-$ ions were 15.0% for $^{137}Cs$(662 keV), 13.1% for $^{54}Mn$(835 keV), and 18.0% and 6.3% for $^{22}Na$(511 keV and 1275 keV), respectively. The decay curves had fast and slow components, and the fast component was about 41 ns independent on the concentration of the $Br^-$ ions. The time resolution of CsI(Br) scintillators decreased with increasing of the concentration of $Br^-$ ions.

  • PDF