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Preparation of Polyethersulfone Ultrafiltration Membranes Containing $ZrO_2$ Nanoparticles by Combining Phase-inversion Method/Sol-gel Technique (상변환/졸-겔법에 의한 $ZrO_2$ 나노입자 함유 Polyethersulfone 한외여과 막의 제조)

  • Youm, Kyung-Ho;Lee, Yun-Jae
    • Membrane Journal
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    • v.16 no.4
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    • pp.303-312
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    • 2006
  • The asymmetric hybrid membranes of polyethersulfone (PES) and $ZrO_2$ nanoparticles were prepared via new one-step procedure combining simultaneously the phase-inversion method and the sol-gel technique. The optimum contents of $Zr(PrO)_4\;and\;HNO_3$ catalyst were determined by the adsorption experiments of phosphate anion onto the resulting hybrid membranes. The maximum adsorption of phosphate anion is obtained at the conditions of 0.15 mL $Zr(PrO)_4$ addition per 1 mL PES and 30 mL $HNO_3$ addition per 1 mL $Zr(PrO)_4$. Variation of morphology, performance and incorporated $ZrO_2$ amount of the resulting hybrid membranes were discussed and determined using SEM, pure water flux, TGA, ICP, XRD and contact angle measurements. Increasing $Zr(PrO)_4$ addition into casting solution, pure water flux is increased and $ZrO_2$ amount in the hybrid membrane is maximized at the conditions 0.15 mL $Zr(PrO)_4$ addition per 1 mL PES. The prephosphatation of PES-$ZrO_2$ hybrid membrane was studied to modify the surface characteristics of membrane. Ultrafiltration of bovine serum albumin (BSA) solution was performed in a dead-end cell using both a bare (non-phosphated) and a phosphated hybrid membrane. It is revealed that both the permeate flux and BSA rejection were increased as about 40% by prephosphatation of hybrid membrane. These results may be explained on the basis of the increase of membrane hydrophilicity, which was determined from contact angle measurements.

Effect of Silkworm Hemolymph Collected Large-scale Bleeding Method in Insect Cell Culture (절피원심법으로 대규모 채취한 누에체액의 곤충세포 증식효과)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Kim, Sam-Eun;Han, Myung-Sae
    • Journal of Sericultural and Entomological Science
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    • v.49 no.1
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    • pp.28-32
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    • 2007
  • This study was carried out to investigate the utilization of hemolymph of silkworm, Bombyx mori, as a substitute for fetal bovine serum(FBS) in the insect cell culture. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate for large scale collection. The hemolymph was collected from 5 th instar larva by centrifugation after cutting of the abdominal legs was more appropriate procedure for large scale collection. The cell growth in the medium supplemented with hemolymph(Baekokjam) collected in large scale was almost same as that in the medium hemolymph supplemented with hemolymph collected in small scale. However, the mutant($wE^b$) hemolymph collected in large scale was still less effective in the cell growth, as compared to the Baekokjam hemolymph collected in large scale. The optimum centrifugation condition for large-scale bleeding was 500 rpm and 15 min.

Selection of Mutant Silkworm with Oxidation-deficient Haemolymph for Insect Cell Culture (곤충세포 배지 개발을 위한 체액산화지연 돌연변이 누에계통 선발)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Yoon, Hyung-Joo;Ahn, Mi-Young;Kim, Sam-Eun;Hwang, Seok-Jo
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.47-50
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    • 2007
  • Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.

Micro-Spot Atmospheric Pressure Plasma Production for the Biomedical Applications

  • Hirata, T.;Tsutsui, C.;Yokoi, Y.;Sakatani, Y.;Mori, A.;Horii, A.;Yamamoto, T.;Taguchi, A.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.44-45
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    • 2010
  • We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

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Effects of follicle size and oocyte diameter on in vitro nuclear maturation of Korean native cattle oocyte (난포크기 및 난자직경과 관련된 한우 체외배양 난자의 핵성숙에 관한 연구)

  • Yong, Hwan-yul;Kim, Hyun-il;Lee, Eun-song;Lee, Byeong-chun;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.929-939
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    • 1996
  • The present study was undertaken to establish a relationship between bovine follicle size and oocyte diameter, compare the nuclear maturation competence of oocytes of different diameter groups and the nuclear maturation changes in Korean Native Cattle according to in vitro maturation period. To compare the relationship between follicle size and oocyte diameter, follicles were dissected, measured, and assigned to one of the following size categories($4{\geq}mm$, 3-4mm, 2-3mm, 1-2mm, and < 1mm), investigate the maturation competence in the different-sized oocytes, which were divided into three groups( < $110{\mu}m$, 110 - < $120{\mu}m$, and ${\geq}120{\mu}m$). Oocytes were cultured in the culture medium during 0, 6, 12, 18, and 24hrs, respectively, stained, and measured the nuclear maturation degree according to period. When compared the relationship between follicle size and intrafollicular oocyte diameter, oocyte diameters of three groups of ${\geq}3mm$ follicle-sized were significantly higher than < 3mm (p<0.01). After in vitro maturation, the rates reached to MI stage of < $110{\mu}m$ oocyte groups(25%) was higher than $110-120{\mu}m$ and ${\geq}120{\mu}m$ oocyte groups(11 and 10%) reached to the same stage(p<0.01), and the rates throughout MII stage of $110-120{\mu}m$ and ${\geq}120{\mu}m$ and < $110{\mu}m$(70 and 76%) groups were higher than < $110{\mu}m$(35%)(p<0.01). When nuclear maturation rates were measured according to period, < 6hr groups(7 and 10%) showed lower rates reached to MI than ${\geq}12hr$ groups(100%), 24hr groups(76%) revealed higher rates throughout MII than 18hr groups(40%). These results indicate that the preparation of oocyte for the production of in vitro fertilization embryos and nuclear transplantation ones could be adapted, as follicle increased up to appointed size there was a corresponding increase in oocyte diameter, and differences of nuclear maturation rate revealed according to oocyte diameter and maturation period.

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Effect of Deep Seawater on Expression of μ-Opioid Receptor in Cultured Rat Hippocampal Neurons (배양된 쥐 해마신경세포에서 μ-아편양 수용체의 발현에 대한 해양심층수의 영향)

  • Moon, Il-Soo;Kim, Seong-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.176-182
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    • 2011
  • Deep seawater (DSW) generally refers to seawater at depths equal to or greater than 200 meters. DSW is rich in inorganic materials which have attracted attention for its various applications. In this study we investigated the effects of the DSW upwelled from the East Sea, offshore Yang Yang (KangWon-do, Korea), on the expression of ${\mu}$-opioid receptor (MOR) of cultured rat hippocampal neurons. Neurons were grown in a minimal essential medium containing 10% (v/v) fetal bovine serum and either 25% (v/v) distilled water, or hardness (H) 800, or H 1000 DSW. Cultures grown in the presence of DSW with H 800 and H 1000 exhibited robust MOR immunoreactive signals in both neurons and astrocytes. Interestingly, the increase in MOR immunoreactive signals was more dramatic in astrocytes than in neurons. Statistical analysis revealed that the relative intensities for MOR clusters increased approximately 4-fold in astrocytes cultured in H 800 and H 1000 media. These increases were statistically very significant (p<0.001). In contrast, the increase in intensities for MOR immunoreactive signals was relatively less dramatic in neurons, where only the increase in the H 1000 culture was statistically very significant (p<0.001). These results indicated that DSW promotes expression of MOR in both neurons and astrocytes, and more significantly in the latter.

Dependences of Ultrasonic Parameters for Osteoporosis Diagnosis on Bone Mineral Density (골다공증 진단을 위한 초음파 변수의 골밀도에 대한 의존성)

  • Hwang, Kyo Seung;Kim, Yoon Mi;Park, Jong Chan;Choi, Min Joo;Lee, Kang Il
    • Journal of the Korean Society for Nondestructive Testing
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    • v.32 no.5
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    • pp.502-508
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    • 2012
  • Quantitative ultrasound technologies for osteoporosis diagnosis measure ultrasonic parameters such as speed of sound(SOS) and normalized broadband ultrasound attenuation(nBUA) in the calcaneus (heel bone). In the present study, the dependences of SOS and nBUA on bone mineral density in the proximal femur with high risk of fracture were investigated by using 20 trabecular bone samples extracted from bovine femurs. SOS and nBUA in the femoral trabecular bone samples were measured by using a transverse transmission method with one matched pair of ultrasonic transducers with a center frequency of 1.0 MHz. SOS and nBUA measured in the 20 trabecular bone samples exhibited high Pearson's correlation coefficients (r) of r = 0.83 and 0.72 with apparent bone density, respectively. The multiple regression analysis with SOS and nBUA as independent variables and apparent bone density as a dependent variable showed that the correlation coefficient r = 0.85 of the multiple linear regression model was higher than those of the simple linear regression model with either parameter SOS or nBUA as an independent variable. These high linear correlations between the ultrasonic parameters and the bone density suggest that the ultrasonic parameters measured in the femur can be useful for predicting the femoral bone mineral density.

Helixor A Inhibits Angiogenesis in vitro Via Upregutation of Thrombospondin-1 (Helixor A는 시험관 내에서 thrombospondin-1의 상승조절을 통해 신혈관생성을 억제한다.)

  • Yeom Dong-Hoon;Hong Kyong-Ja
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.895-903
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    • 2005
  • Thrombospondin-1 (TSP-1), a negative regulator in tumor growth and angiogenesis, is cell-type specifically regulated under pathological conditions or by extracellular stimuli, and the regulation of TSP-1 gene expression is important for developing new approaches in tumor therapy. Mistletoe is a parasitir plant that have been used for immunomodulation and antitumor therapy. Helixor A is an aqueous part of mistletoes extract. Here we showed that TSP-1 expression was significantly induced at both mRNA and protein levels in the Hepatocarcinorna cell line (Hep3B) and primary bovine endothelial cell line (BAE) exposed to Helixor A. Our promoter analysis confirmed that the expression of TSP-1 gene was regulated by Helixor A at the transcriptional level. In cell invasion assay, the conditioned media obtained from treatment of these cells significantly reduced the number of invasive cells and also inhibited capillary-like tube formation of BAE cells on Matrigel. Moreover, the inhibitory efforts of the conditioned media on cell invasion and tube formation were reversed by blocking with anti-TSP-1 neutralizing antibodies, suggesting that TSP-1 is involved in Helixor A-indured antiangiogenic effect. Taken together, our results suggest that Helixor A have an antiangiogenic effects through upregulation of TSP-1.

Effect of storage conditions of olive flounder Paralichthys olivaceus serum on enzyme-linked immunosorbent assay (넙치 혈청의 보존 조건이 ELISA 결과에 미치는 영향)

  • Kim, Wi-Sik;Jang, Min-Seok;Kim, Jong-Oh;Kim, Du-Woon;Jung, Sung-Ju;Kim, Seok-Ryel;Park, Myoung-Ae;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.22 no.2
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    • pp.167-172
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    • 2009
  • Antibody-detection enzyme-linked immunosorbent assay (ELISA) was performed to determine whether the absorbance value of ELISA is influenced by the different serum storage conditions of olive flounder Paralichthys olivaceus. Flounder antiserum to bovine serum albumin was stored at -80, -20, 4 and 20${^{\circ}C}$ for 1, 30, 69 and 124 days, respectively. In addition, the flounder antisera were frozen at -80 and -20${^{\circ}C}$, respectively and then repeated 1, 5 and 10 thaw-freeze cycles. No significant difference was shown in ELISA optical density (O.D.) values of sera, which were stored at the above mentioned storage conditions during 124 days. ELISA O.D. values of unfrozen serum samples, which were previously stored at 4${^{\circ}C}$, were almost similar to those of sera undergoing 1, 5, and 10 freeze-thaw cycles after stored at -80 or -20${^{\circ}C}$. In conclusion, the ELISA O.D. values of flounder sera were not affected by various storage conditions: different temperatures (-80, -20, 4 and 20${^{\circ}C}$), durations of storage (1, 30, 69 and 124 days), and repeated thaw-freeze cycles (1, 5, and 10 times).

Effects of Antioxidative stress, Antimutagenicity and Cytotoxicity of Cancer cells in Fractional Extracts from Ganoderma lucidum Karst (영지버섯(Ganoderma lucidum Krast)의 순차 분획물의 산화적 스트레스 억제효과와 항돌연변이 및 암세포 증식 억제 효과)

  • Oh Se-In;Lee Mee-Sook
    • Korean journal of food and cookery science
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    • v.21 no.6 s.90
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    • pp.759-768
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    • 2005
  • This study was performed to determine the antioxidative effect of the hexane, dichloromethane, ethylacetate, butanol and water fractions of Ganoderma lucidum extracts on the inhibition of malondialdehyde(MDA) and bovine serum albumin(BSA) conjugation reaction, the inhibition of lipid peroxidation and the scavenging effect on 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical, the antimutagenic capacity as measured by the Ames test and the inhibitory effect on cancer cell. Ganoderma lucidum is believed to have possible antioxidative capacities, although the results have varied according to the assay method. The most effective antioxidative capacity was inhibition of lipid peroxidation. Among the five fractions, water fraction showed strong inhibition rates on MDA & BSA conjugation reaction, and ethylacetate fractions showed the most effective inhibition rate on lipid peroxidation and scavenging effect on DPPH radical. The indirect and direct antimutagenic effects of ethanol extracts of Ganoderma lucidum were examined by Ames test using Salmonella typhimurium TA98 and TA100. Among the samples, the water fraction did not have any antimutagenic effect. The inhibition rates on mutagenicity in the presence of 2.5 mg/plate were nearly $100\%$ for Salmonella typhimurium TA98 and TA100 except the hexane fraction of the direct mutagenicity mediated by 2-Nitrofluorene in Salmonella typimurium TA98($64.69\%$). Under the 2.5 mg/plate concentration, the inhibitory effects of hexane and dichloromethane fraction were superior to that of the other fractions on the direct mutagenicity for Salmonella typhimurium TA100 and indirect mutagenicity for Salmonella typhimurium TA98 and TA100. The inhibitory effect of Ganoderma lucidum extracts on cell proliferation in HeLa and MCF-7 was investigated by U test. The dichloromethane fraction showed highly antiproliferative effect in HeLa and MCF-7($IC_{50}$: 0.122 mg/mL, 0.272 mg/mL, respectively) cells while the water faction had a weak inhibitory effect($IC_{50}$: 0.691 mg/mL, 10.919 mg/mL respectively). These results suggest that Ganoderma lucidum may have antioxidative, antimutagenic and anticancer capacities and may be a candidate of the prevention and dietetic treatment of chronic diseases and the development of antioxidative, antimutagenic and anticancer functional food.