• Title/Summary/Keyword: modified method

Search Result 8,761, Processing Time 0.043 seconds

A Reliable Protocol for transfection of mature primary hippocampal neurons using a neuron-glia co-culture system (신경세포-신경교세포 공동배양을 이용한 성숙한 해마신경세포의 효율적인 형질전환 방법)

  • Lee, Hyun-Sook;Cho, Sun-Jung;Jung, Yong-Wook;Jin, Ing-Nyol;Moon, Il-Soo
    • Journal of Life Science
    • /
    • v.17 no.2 s.82
    • /
    • pp.198-203
    • /
    • 2007
  • DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

Comparative Studies on the Enzymatic Properties of Trypsins from Cat-shark and Mackerel -1. Purifications and Reaction Conditions of the Trypsins- (복상어와 고등어의 Trypsin에 관한 비교 효소학적 연구 -1. Trypsin의 정제와 반응조건-)

  • PYEUN Jae-Hyeung;CHO Deuk-Moon;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.24 no.5
    • /
    • pp.273-288
    • /
    • 1991
  • To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

  • PDF

Effect of Paroxetine and Sertraline Treatment on Forced Swim Test-Induced Behavioral and Immune Changes in the Mouse (마우스 강제수영에 의한 행동 및 면역반응 변화에 대한 Paroxetine과 Sertraline의 효과)

  • Eum, Se-Yeun;Jeong, Min-Ho;Lim, Young-Jin;Kim, Bu-Kyung;Jeong, Soo-Jin;Hahn, Hong-Moo;Choe, Byeong-Moo
    • Korean Journal of Psychosomatic Medicine
    • /
    • v.8 no.1
    • /
    • pp.46-57
    • /
    • 2000
  • Objectives : The purpose of the present study was to examine the effect of subacute treatment with the selective serotonin reuptake inhibitors(paroxetine and sertraline) on immobility in the forced swim test(FST) and on FST-induced changes in immune parameters of the mice. Methods : Authors applied a modified method of FST by Porsolt et al. Over 5 BALB/c mice were used for each group of experiments. To explore the changes in immune parameters by FST, authors investigated the production of anti-rat RBC antibody, concanavalin A(ConA)- or lipopolysaccharide(LPS)-stimulated splenocytes proliferation assay and cytokine gene expression. Results : Both paroxetine and sertraline decreased the duration of immobility in a dose-related manner. FST-performed mice showed a significant decrease in mitogenic responses of splenocytes and a slight increasing tendency in anti-rat RBC antibody response. All these responses were attenuated significantly by paroxetine and attenuated nearly nominal significance level by sertraline. The cytokine profiles of ConA-stimulated splenocytes from FST-performed mice showed stronger expression of IL-4 and weaker expression of IL-2 than control mice, and no changes in the expressions of IFN-$\gamma$ and lymphotoxin. IL-6 and IL-10 were not expressed in both group of mice. The pretreatment of paroxetine and sertraline attenuated the altered cytokine expressions in FST-performed mice to some extent. Some alterations of the expressions of IL-6 and IL-10 were observed in the mice which the selective serotonin reuptake inhibitors had been pretreated. Conclusion : The subacute treatment of paroxetine and sertraline attenuated the FST-induced behavioral and immune changes, and these serotonin reuptake inhibitors may exert some modulating effects on the immune system by the induction of cytokine gene expression, especially IL-6 and IL-10.

  • PDF

Results of Postoperative Radiation Therapy of Rectal Cancers - with the Emphasis of the Overall Treatment Time - (직장암의 수술 후 방사선치료의 성적 - 예후 인자와 전체 치료기간이 미치는 영향에 관한 고찰 -)

  • Kim Joo-Young;Lee Myung-Hag;Lee Kyu-Chan
    • Radiation Oncology Journal
    • /
    • v.16 no.3
    • /
    • pp.303-310
    • /
    • 1998
  • Purpose : To evaluate the results of the treatment of locally advanced but resectable rectal cancers and to analyze prognostic factors. especially with the emphasis on the treatment time factor. Materials and Methods : There were 71 patients with rectal cancer who had been treated by curative surgical procedure and postoperative radiotherapy from August 1989 to December 1993. The minimum follow up period was 24 months and the median follow-up was 35 months Radiation therapy had been given by 6 MV linear accelerator by parallel opposing or four-box portals. Whole pelvis was treated up to 5040 cGy in most cases. Systemic chemotherapy had been given in 94$\%$ of the patients, mostly with 5-FU/ACNU regimen. Assessment for the overall and disease-free survival rates were done by life-table method and prognostic factors by Log-Rank tests. Results : Five-year overall survival, disease-free survival were 58.8$\%$ and 57$\%$, respectively. Two-year local control rate was 76.6$\%$. Stage according to Modified Astler-Coller (MAC) system, over 4 positive lymph nodes, over 6weeks interval between definitive surgery and adjuvant radiotherapy and over 7 days of interruption during radiotherapy period were statistically significant, or borderline significant prognostic factors. Conclusion : The treatment results of patients with rectal cancers are comparable to those of other large institutes. The treatment results for the patients with bowel wall penetration and/or positive regional lymph nodes were still discouraging for their high local recurrence rate for the patients with MAC 'c' stage diseases and high distant metastases rate even for the patients with node-negative diseases. Maybe more effective regimen of chemotherapy would be needed with proper route and schedule. To maximize postoperative adjuvant treatment. radiotherapy should be started at least within 6 weeks after surgery and preferably as soon as wound healing is completed. Interruption of treatment during radiotherapy course affects disease-free survival badly, especially if exceeds 7 days. So, the total treatment period trout definitive surgery to the completion of radiotherapy should be kept as minimal as possiable.

  • PDF

A Study on Contents of Vitamin K1 in Local Agricultural Products (지역농산물의 비타민 K1 함량 조사)

  • Lee, A Reum;Kim, Joo Hee;Park, Jae-Ho;Kim, Youngho;Hong, Eui Yon;Kim, Haeng-Ran;Choi, Youngmin;Lee, Junsoo;Eom, Hyun-Ju
    • The Korean Journal of Food And Nutrition
    • /
    • v.29 no.3
    • /
    • pp.301-306
    • /
    • 2016
  • Vitamin K (phylloquinone) is an essential cofactor in the synthesis of active blood-clotting factors II, VII, IX and X. Deficiency of vitamin K leads to inadequate activity of these factors, resulting in bleeding. In this study, we investigated vitamin $K_1$ content of agricultural products that are widely and specifically grown in Korea including 9 leaves and vegetables, 16 fruits, and 11 cereals and specialty crops. Vitamin $K_1$ analysis of the agro-samples was by a validated, modified, reversed phase-HPLC method with fluorescence detection after post-column derivatization. The vitamin $K_1$ content ranged from 1.83 to $682.73{\mu}g$/100 g in leaves and vegetables, 0.17 to $28.22{\mu}g$/100 g in fruits, and ND to $279{\mu}g$/100 g in cereals and specialty crops. Among the 36 samples, high content of vitamin $K_1$ were found in Gugija (Lycium chinense Miller) leaves (average $682.73{\mu}g$/100 g) and Hansan ramie leaves (average $423.12{\mu}g$/100 g); however, mushroom, amaranth and Chinese artichoke showed no detectable levels. The results of ourstudy provide reliable vitamin $K_1$ content of Korean grown agricultural products that expand nutritional information and food composition database.

Package Atmosphere and Quality as Affected by Modified Atmosphere Conditions of Persimmon (Diospyros kaki. cv. Fuyu) Fruits ('부유' 단감의 MA포장 조건에 따른 포장기체조성 및 과실의 품질 변화)

  • Ahn, Gwang-Hwan;Song, Won-Doo;Park, Doo-Sang;Lee, Yeon;Lee, Dong-Sun;Choi, Seong-Jin
    • Korean Journal of Food Science and Technology
    • /
    • v.33 no.2
    • /
    • pp.200-204
    • /
    • 2001
  • Persimmon (Diospyros kaki. cv. Fuyu) fruits were packaged under different conditions, and then stored at $0^{\circ}C$ for 21 weeks. The tried packages were heat-sealed bag of one fruit, heat-sealed bag of five fruits, and tie-sealed bag of five fruits, which used films of three different thickness (0.03 ㎜, 0.05 ㎜ and 0.06 ㎜). Throughout the storage, package free volume, package atmosphere and quality were measured. Package free volume decreased with time with higher rate for heat-sealed bags, in which close contact between fruit and the film was eventually reached in longer storage. However, tie-sealed bags maintained the levels of stabilized free volume. The rate of free volume decrease was faster with thinner film and larger bag size for the packs sealed by the same method. Package atmosphere covered $O_2$ concentration of $1.1{\sim}17.1%$ and $CO_2$ concentration of $1.1{\sim}8.3%$, $O_2$ concentration decreased during storage, whereas $CO_2$ increased. Thinner film package created the internal atmosphere of higher $O_2$ and lower $CO_2$ concentrations. Tie-sealed bags of 5 fruits in the films of 0.05 mm and 0.06 mm thickness maintained the equilibrated package atmosphere of $1.1{\sim}3.0%\;O_2$ and $4.0{\sim}5.5%\;CO_2$, which preserved the fruits best in terms of firmness and less physiological changes of flesh browning and peel browning. At five fruit heat-sealed bag in the films of 0.06 mm thickness with $CO_2\;8.3%$, flesh browning occurred at a high rate, whereas all treatments in the film of 0.03 mm thickness with high $O_2$ and relatively low $CO_2$ contents, flesh and peel browning rates were high.

  • PDF

Electrochemical Characterization of Anti-Corrosion Film Coated Metal Conditioner Surfaces for Tungsten CMP Applications (텅스텐 화학적-기계적 연마 공정에서 부식방지막이 증착된 금속 컨디셔너 표면의 전기화학적 특성평가)

  • Cho, Byoung-Jun;Kwon, Tae-Young;Kim, Hyuk-Min;Venkatesh, Prasanna;Park, Moon-Seok;Park, Jin-Goo
    • Journal of the Microelectronics and Packaging Society
    • /
    • v.19 no.1
    • /
    • pp.61-66
    • /
    • 2012
  • Chemical Mechanical Planarization (CMP) is a polishing process used in the microelectronic fabrication industries to achieve a globally planar wafer surface for the manufacturing of integrated circuits. Pad conditioning plays an important role in the CMP process to maintain a material removal rate (MRR) and its uniformity. For metal CMP process, highly acidic slurry containing strong oxidizer is being used. It would affect the conditioner surface which normally made of metal such as Nickel and its alloy. If conditioner surface is corroded, diamonds on the conditioner surface would be fallen out from the surface. Because of this phenomenon, not only life time of conditioners is decreased, but also more scratches are generated. To protect the conditioners from corrosion, thin organic film deposition on the metal surface is suggested without requiring current conditioner manufacturing process. To prepare the anti-corrosion film on metal conditioner surface, vapor SAM (self-assembled monolayer) and FC (Fluorocarbon) -CVD (SRN-504, Sorona, Korea) films were prepared on both nickel and nickel alloy surfaces. Vapor SAM method was used for SAM deposition using both Dodecanethiol (DT) and Perfluoroctyltrichloro silane (FOTS). FC films were prepared in different thickness of 10 nm, 50 nm and 100 nm on conditioner surfaces. Electrochemical analysis such as potentiodynamic polarization and impedance, and contact angle measurements were carried out to evaluate the coating characteristics. Impedance data was analyzed by an electrical equivalent circuit model. The observed contact angle is higher than 90o after thin film deposition, which confirms that the coatings deposited on the surfaces are densely packed. The results of potentiodynamic polarization and the impedance show that modified surfaces have better performance than bare metal surfaces which could be applied to increase the life time and reliability of conditioner during W CMP.

A COMPARATIVE STUDY ON THE CHEMICAL COMPOSITION AND MECHANICAL PROPERTIES OF FOUR LOW-GOLD-CONTENT DENTAL CASTING ALLOYS MANUFACTURED IN KOREA (한국산(韓國産) 치과주조용(齒科鑄造用) 저금함유합금(低金含有合金)의 조성(組成) 및 기계적(機械的) 성질(性質)에 관(關)한 비교연구(比較硏究))

  • Chang, Ik-Tae;Yang, Jae-Ho;Kim, Chang-Whe;Kim, Kwang-Nam;Lee, Sun-Hyung;Kim, Yung-Soo;Chang, Wan-Shik
    • The Journal of Korean Academy of Prosthodontics
    • /
    • v.19 no.1
    • /
    • pp.17-27
    • /
    • 1981
  • This study was conducted to determine the chemical composition and the mechanical properties of four commercially available low gold-based crown and bridge alloy produced in Korea. Four dental casting gold-silver-palladium alloys, i.e., A, B, C and D (code of alloys) were selected for the evaluation of chemical composition, ultimate tensile strength, elongation. values and Vickers hardness. The chemical composition of test specimens was analyzed by both emission spectrography and wet gravitation method with a 1.5gm of low gold ingot. The tensile properties and Vickers hardness was determined with cast specimens treated in following three conditions; as-cast, softening heat treatment and hardening heat treatment. The tensile testing bars were cast in accordance with the model designed by Gettleman and Harrison (1969) which was modified from the A. D. A. Specification No. 14 for dental chromium-cobalt casting alloy. Nine tensile test specimens were made from a split silicone mold for each of the test alloys to the size of 2.5mm in diameter and a gauge length of 10mm. All four alloys were handled in accordance with conventional methods used in Type III gold alloys. Ultimate tensile strength and elongation were measured on an Instron Universal Tensile Testing Machine (Model 1125, Japan) operated at a crosshead rate of 0.1cm/min. Elongation values were measured using Digital Measuring Microscope (MS-152, FUSOH, Japan). Vickers hardness was determined with a Vickers Hardness Tester (Model VKH-l, Japan) at a 1.0kg load on a mounted tensile test specimen. The following results were obtained from this study; 1. All tested alloys were composed of Au, Ag, Pd, Cu, Zn and Fe in common. The composition rate of gold for all four alloys was found in the range of $42{\sim}47$ weight % as shown below. Alloy A; Au 45%, Ag 40.2%, Pd 5.76%, others 9.04%. Alloy B; Au 47.1%, Ag 29.03%, Pd 6.98%, others 16.92%. Alloy C; Au 45%, .Ag 26.9%, Pd 6.83%, others 21.07%. Alloy D; Au 41.8%, Ag 34.4%, Pd 6.95%, others 16.85%. 3. The ultimate tensile strength of the four alloys was in the range of $31{\sim}82kg/mm^2$. The test results were shown in the below order from the highest value; As-cast condition; D, B, C, A. Softening heat treament; B, C, D, A. Hardening heat treatment; D, B, C, A. 4. The test :results of the elongation rate for each alloy were in the range of $0.5{\sim}18%$. The test results were shown in the below order from the highest value; As-cast condition; A, D, B, C. Softening heat treatment; A, C, D, B. Hardening heat treatment; C, D, B, A. 5. Vickers hardness for each of the four alloys was in the range of $120{\sim}230$. The test results were shown in the below order from the highest value; As-cast condition; C, B, D, A Softening heat treatment; D, B, C, A. Hardening heat treatment; D, A, C, B. 6. There were no differences in the physical properties between as-cast condition and softening heat treatment.

  • PDF

A Fluid Analysis Study on Centrifugal Pump Performance Improvement by Impeller Modification (원심펌프 회전차 Modification시 성능개선에 관한 유동해석 연구)

  • Lee, A-Yeong;Jang, Hyun-Jun;Lee, Jin-Woo;Cho, Won-Jeong
    • Journal of the Korean Institute of Gas
    • /
    • v.24 no.2
    • /
    • pp.1-8
    • /
    • 2020
  • Centrifugal pump is a facility that transfers energy to fluid through centrifugal force, which is usually generated by rotating the impeller at high speed, and is a major process facility used in many LNG production bases such as vaporization seawater pump, industrial water and fire extinguishing pump using seawater. to be. Currently, pumps in LNG plant sites are subject to operating conditions that vary depending on the amount of supply desired by the customer for a long period of time. Pumps in particular occupy a large part of the consumption strategy at the plant site, and if the optimum operation condition is not available, it can incur enormous energy loss in long term plant operation. In order to solve this problem, it is necessary to identify the performance deterioration factor through the flow analysis and the result analysis according to the fluctuations of the pump's operating conditions and to determine the optimal operation efficiency. In order to evaluate operation efficiency through experimental techniques, considerable time and cost are incurred, such as on-site operating conditions and manufacturing of experimental equipment. If the performance of the pump is not suitable for the site, and the performance of the pump needs to be reduced, a method of changing the rotation speed or using a special liquid containing high viscosity or solids is used. Especially, in order to prevent disruptions in the operation of LNG production bases, a technology is required to satisfy the required performance conditions by processing the existing impeller of the pump within a short time. Therefore, in this study, the rotation difference of the pump was applied to the ANSYS CFX program by applying the modified 3D modeling shape. In addition, the results obtained from the flow analysis and the curve fitting toolbox of the MATLAB program were analyzed numerically to verify the outer diameter correction theory.

The Bibliographical Investigation of Sasang Constitution Diagnosis (사상체질(四象體質) 진단법(診斷法)의 문헌적(文獻的) 고찰(考察) - 외형(外形), 심성(心性), 증(證)을 위주(爲主)로 -)

  • Jung, Won-gyo;Kim, Jong-weon
    • Journal of Sasang Constitutional Medicine
    • /
    • v.11 no.2
    • /
    • pp.95-117
    • /
    • 1999
  • 1. Purpose of study There are many Constitution theories. But, In the Sasang Constitutional medicine, the theory and the practice are connected systematically. So it has been used in Korea popularly. Lee Je-Ma suggested that the external form, the mind, the symptom are the criteria of Sasang constitution diagnosis at the Discourse on the Four Principle and the Discourse on the Identifying Four Constitutions. But The clinical importance study of three diagnostic criteria - the external form, the mind, the symptom - has never been studied. Therefore, there are many obstacles of using the three diagnostic criteria totally. And I think that it needs a study which we apply the three diagnostic criteria to practice and approve the clinical importance of one another. I classify and investigate the previous methods of Sasang constitution diagnosis by the three diagnostic criteria in this study. 2. Method of study I investigate the previous methods of Sasang constitution diagnosis bibliographically by Dongyi Soose Bowon, Journal of Constitutional medicine and other books. 3. Result of study (1) Study of the external form The Study of Ki-sang(氣像) and Sa-ki(詞氣) by experience and intuition was done previously, and study of imaginay formulation of each constitution by measuring Chae-hyung(體形) and yong-mo(容貌) has being done recently. (2) Study of the mind For the objectification of study and diagnosis, self-reporting questionnaire has been used. The ideal choice to determine an indivisual constitution, as far as questionnaire go, is to take results from both the modified version Sasang Pattern Identification Questionnaire and QSCCII. (3) Study of the symptom For the study of the symptom - constitution symptom and constitution symptom of disease - the study of principles, the clinical study of the type of disease and symptom, the study which the pathologic view of Oriental medicine apply the view of Sasang Constitutional medicine have been done. We must try to objectfy Sasang Constitutional disease.

  • PDF