• Korean Chemical Engineering Research
• /
• v.55 no.3
• /
• pp.369-378
• /
• 2017

Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

• Korean journal of applied entomology
• /
• v.43 no.3 s.136
• /
• pp.225-231
• /
• 2004

An endoparasitoid wasp, Cotesia plutellae, has been used for a biological control agent against the diamondback moth, Plutellae xylostella. It has a symbiotic polydnavirus in their reproductive tract, which is required for its successful parasitization. Here, we measured a specific replication time of the polydnavirus during female development of C. plutellae. We, also, analyzed the reproductive potentials of female C. plutellae under mating or different host conditions. At $25^{\circ}C$, pupal C. plutellae began to develop adult tissues such as compound eyes and wings since day 2. At day 5, all adult tissues including antennae were developed and were ready to emerge. With polyclonal antibody raised against C. plutellae polydnavirus, an immunobloting could confirm virus replication at day 4 during pupal stage. Virus particles could be visualized by transmission electron microscope in the oviduct lumen of day 5 pupae. After adult eclosion, venom gland and ovarian calyx increased in size, though ovarioles did not. Mated females layed large number of eggs (over $60\%$) at first 4 days during their mean longevity of ca. 8 days at $25^{\circ}C$. Unmated females showed less active ovipositional behavior, where all the eggs developed into males. C. Plutellae parasitized both P. xylostella and fall webworm, Hyphantria cunea. However, C. Plutellae developed faster and showed higher successful paarasitization in P. xylostella than in H. cunea.

• Composites Research
• /
• v.29 no.4
• /
• pp.125-131
• /
• 2016

In case of CNT mixing with epoxy, epoxy matrix needs to be diluted. This work studied the effect of the dilution condition of epoxy on CNT dispersion. The optimum solvent condition using acetone and DMF was found via mechanical and solubility methods which affects, the epoxy performance. The dispersion using acetone was better than the DMF and thus higher mechanical properties. Four mixing types of CNT particle were performed. To verify the effects of each step between dilution and dispersion, the dispersion between epoxy and CNT was evaluated via the electrical resistance and optical methods. The optimum dispersion was obtained via mechanical test and thermal analysis by DSC. Among four types, the best was to disperse CNT after epoxy and hardeners were diluted respectively.

• Asian Journal of Turfgrass Science
• /
• v.20 no.1
• /
• pp.83-91
• /
• 2006

This research was initiated to enhance the tilth of fine-textured soil for turf growth by incorporation of crumb rubber shredded from used tires. A specific objective was to determine the physical properties of soil mixes amended with different grade and amount of crumb rubber in soils. Two soils and three different grades(3.5, 6.5 and 9.5 mm) of crumb rubber were used. The soils selected were an Arenzville silt loam(coarse-silty, mixed, nonacid, mesic Typic Udifluvents) and a Hosmer silty clay loam(fine-silty, mixed, mesic Typic Fragiudalfs). The amount of crumb rubber mixed in soil ranged from 0 to 0.4 $g{\cdot}g^{-1}$(using 0.05 $g{\cdot}g^{-1}$) increments and 0 as a control. For each treatment, soil cores were constructed following the recommendation by the United States Golf Association Green Section Record. Results indicated that porosity of the mixes decreased as the amount of crumb rubber increased. Regardless of the grade of crumb rubber, mixes with less than 0.15 $g{\cdot}g^{-1}$ of crumb rubber in fine-textured soil could not enhance their macro-porosity and hydraulic conductivity. However, as the amendment increased over 0.15 $g{\cdot}g^{-1}$, the tilth of the mixes had improved significantly macro-porosity, hydraulic conductivity and air permeability, as compared with a control.

• Journal of the Korean Chemical Society
• /
• v.59 no.1
• /
• pp.22-30
• /
• 2015

The effect of diphtheria toxin on cell membrane lipids was studied by examining the phospholipase D (PLD) activity and free fatty acids (FFA) release in HepG2 cells. The diphtheria toxin effects on lipid alteration show apparently maximal at pH 5.1, stimulating PLD activity nearly 3.5 fold and enhancing FFA release approximately 5 fold over the control. These results indicate that the membrane is perturbed and its lipid component is rearranged during the diphtheria toxin translocation. Digitonin, a random membrane perturbing detergent, exhibit about four-fold higher perturbation effect over the diphtheria toxin at neutral pH. This observation suggests that the membrane perturbation induced by diphtheria toxin appears to be rather selective. To investigate the cause of the membrane perturbation, Cibacron blue, an inhibitor of membrane pore formation, and hemagglutinin, an influenza virus with fusion peptide, were tested for their effects on diphtheria toxin action. Cibacron blue decreased the diphtheria toxin effect by almost 50%, but the lipid alteration induced by hemagglutinin was similar to the diphtheria toxin effect. These observations imply that the membrane perturbation induced by diphtheria toxin may be caused by a combination of pore formation and insertion of hydrophobic peptide of toxin to the membrane as well. Additionally, we found that the diphtheria toxin increased the HepG2 cells permeability but the cells viability was maintained at high level at the same time. DNA fragmentation which is related to apoptosis was not induced by the toxin. Under these conditions, we could demonstrate that the lipid alteration of HepG2 cells was brought about by diphtheria toxin at acidic pH.

• Applied Microscopy
• /
• v.37 no.4
• /
• pp.249-258
• /
• 2007

The Pancreatic islet are the clusters of endocrine cells scattered through out the exocrine pancreas. Transplantation of a sufficient pancreatic islets can normalize blood glucose level so that may prevent devastating complications of type I diabetes(IDDM) and other side effects of the IDDM. Recently, there are several approaches to transplant sufficient pancreatic islet, and it was comprised in increase or regeneration of the endogenous $\beta$-cell mass from donor's pancreas, but relatively few studies have been devoted to the morphological characters of the isolated and 3 day cultured pancreatic islets. We investigated morphological pattern of intracellular structure of isolated and 3 day cultured pancreatic islets. The morphological characters of the pancreatic islets were observed by scanning electron microscope and transmission electron microscope, and insulin distribution of the each islets were observed by transmission electron microscope, and were labeled with insulin antibody. Intracellular structures including nuclei, mitochondria, RER, Golgi complex and many secretory granules were normally appeared in the isolated pancreatic islets which was extracted immediately dornor's pancreas, however, There is a significant morphological changes between the 3 day cultured pancreatic islets and isolated islets. 3 day cultured pancreatic islet's $\beta$-cells had normal nuclei but increased cytoplasm mass and RER and developed Golgi complex. Insulin secretory granules were decreased in numbers rather than isolated pancreatic islet. In this study, the pattern of intracellular structure variation was examined during pancreatic islet culture. Most distinct features are variation of the insulin secretory granules, and developed RER, and dilated golgi complex. Therefore, we suggested that the various change of the morphological characters on cultured pancreatic islets were responsible for the function(biosynthesis and secretion of insulin) and growth. These results were also cultured islets have greater ability to recover and maintain normoglycemia than isolated islet transplantation.

• Journal of Soil and Groundwater Environment
• /
• v.7 no.3
• /
• pp.85-94
• /
• 2002

Overall objective of this study was to evaluate the electrochemical characteristics of fine soils during the electrokinetic(EK) remediation. Zeta potential of kaolinite as a function of solution pH and surfactant concentration was investigated to make a relationship with electroosmotic flow direction and rate. During the EK experiments, pH of pore solution, electroosmotic permeability($k_e$), electric conductivity($\delta_e$) and voltage distribution was measured, respectively, The point of zero charge(PZC) of kaolinite was estimated to be about 4.2 and the zeta potential of kaolinite above PZC was more negative as solution pH increased. Sorption of surfactants on the kaolinite altered the zeta potential of kaolinite. resulting from the variation of electrochemical characteristics of kaolinite surface. hs the EK experiment progressed, low pH was predominant over most of the kaolinite specimen and thus resulted in very low mass and charge flow. The $k_e$ and $\delta_e$ was also affected by the variation of voltage drop across the EK column with time. Results from this study implied that zeta potential of kaolinite affected by the pH variation of pore solution and voltage distribution in soil column played important role in the determination of mass and charge flow during EK process. It was also suggested that pH adjustment or addition of suitable sorbates could alter the electrochemical characteristics of soil surface and thus maintain high mass and charge flow rate with time.

• Journal of fish pathology
• /
• v.11 no.2
• /
• pp.129-136
• /
• 1998

In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

• Journal of Yeungnam Medical Science
• /
• v.23 no.2
• /
• pp.182-192
• /
• 2006

Background: Atherosclerosis has emerged as the leading cause of death in developed countries. At present, human umbilical vein endothelial cells (HUVEC) are most commonly used for the investigation of Endothelial cells (EC). However, HUVEC are not found in arteries but only in veins. Currently there are many reports on methods used to isolate EC;, most of these methods require special equipment to remove contaminating smooth muscle cells (SMC). Materials and Methods: The method described here may be used to isolate not only ECs but also SMCs;,the approach presented here did not require special equipment. Rat aorta was treated with 2 mg/ml of type II collagenase solution for 45 minutes. The isolated cells from the aorta were incubated in medium G for a week;, only ECs could be separated. After the collagenase treatment, the rest of aorta was cut lengthwise, and left undisturbed to obtain SMCs in the culture dish for 10 days. To verify the purity of the isolated cells, we performed immunofluorescence and evaluated the results with transmission electron microscopy analysis. Results: The immunofluorescence study demonstrated specific expression of CD31 and ${\alpha}$-smooth muscle actin in the isolated ECs and SMCs, respectively. Cultured ECs and SMCs showed their own fine structure characteristics. Conclusion: These results suggest that this method for isolating ECs and SMCs may be especially useful for the study of atherosclerosis.

• Polymer(Korea)
• /
• v.28 no.2
• /
• pp.135-142
• /
• 2004

Poly(vinyl acetate) (PVAc) prepared by emulsion polymerization has broad applications for additives such as paint binder, adhesive for wood and paper due to its low glass transition temperature which help to plasticize substrate resins. Since emulsion polymerization has a disadvantage that surfactant and ionic initiator degrade properties of the product polymer, poly(vinyl acetate-co-butyl acrylate) (VVc-BA) was synthesized using potassium persulfate as catalyst and poly(vinyl alcohol) (PVA) as protective colloid to prevent the degradation. The copolymer latex product was internally plasticized and has enhanced colloid stability, adhesion, tensile strength and elongation. During VAc-BA emulsion polymerization, no coagulation and complete conversion occur with the reactant mixture of 0.7wt% potassium persulfate, 15wt% poly(vinyl alcohol) (PVA-217), and the balanced monomer that the weight ratio of vinyl acetate to butyl acrylate is 19. As the concentrations of PVA increase, the copolymerization becomes faster and polymer particles are more stable, resulting in enhanced mechanical stability of the VAc-BA copolymer. However, the size of the polymer particles decreases with increasing PVA contents. Properties of the VAc-BA copolymer, such as minimum film formation temperature, glass transition temperature, surface morphology, molecular weight and molecular weight distribution, tensile strength and elongation, were characterized using differential scanning calorimeter, transmission electron microscope and other instruments.