• Title/Summary/Keyword: 24-hour pH

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Peptide Production from the Washing Liquid of the Fish Paste of Alaska pollak (Theragria chalcogramma) by Immobilized Enzyme (고정화 효소를 이용한 명태고기풀 수세액으로부터 Peptide 생산에 관한 연구)

  • SHIN Suk-U;SUETSUNA Kunio
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.466-472
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    • 1997
  • Peptides separated from fish paste washing liquid of an Alaska pollak (Theragria chalcogramma) were purified and characterized. The fish paste washing liquid (supernatant) was separated by centrifugation of fish paste homogenate. The fish paste washing liquid of $0.5\%$ concentration was hydrolyzed for 24 hour at $50^{\circ}C$ by immobilized protease in bioreactor and decomposing liquid of protein having $50\%$ decomposing rate (OPA method) was obtained. The crude peptide fractions were obtained from this liquid by Dowex 50w $(H^+)$ column chromatograpy. Purified peptides (SP-fraction peptides) were fractionated by using SP-Sepadex C-25 $(H^+)$ column chromatography. Molecular weights and amino acid compositions of these peptides were estimated by Sephadex G-50 column chromatography and HPLC, respectively. when the washed peptides was eluated with $0.6\~0.9\%\;and\;1.2\~2.0\%$ of NaCl, peptides composed of weakly basic amino acids and strongly basic amino acid were respectively eluted. Molecular weights of each peptide fractions showed the broad distribution from 1,000 Da to 3,000 Da in the order of SP-4>SP-3>SP-2>SP-1. Peptides contained a large quantity of glycine, arginine, glutamic acid, and alanine in the washed peptide and its SP-tractions, respectively.

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Spectrophotometric Determination of Aluminium Ion in Drinking Water by Flow Injection Analysis (흐름주입분석법에 의한 음용수 중 알루미늄 이온의 분광광도법 정량)

  • Choi, Yong-Wook;Jin, Jae-Young
    • Journal of the Korean Chemical Society
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    • v.44 no.5
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    • pp.422-428
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    • 2000
  • Optimum analytical conditions of the aluminium ion were established by flow injection analysis. Eriochrome Cyanine R(ECR) dye reacts with the aluminium ion at pH 6.0 to form a complex that exhibits maximum absorption at 535 nm. Reaction conditions including the mixing and the reaction coil length, the concentration and the pH of the buffer solutio, temperature, and injection loop volume were optimized to intro-duce this reaction into flow injection analysis. The results were as follows. A mixing coil length of 0.5 m and a reaction coil length of 4.0 m, the pH 6.0 and 1M of acetate buffer solution, the ECR concentration of 0.56 mM, the reaction temperature of 40$^{\circ}C$, the injection loop volume of 300${\mu}L$ were chosen as optimum conditions. Under these conditions the detection limit of the aluminiumion was less than 0.05 mg/L and the repeatability was better than 1%. A sampling frequency of 24 times for an hour was achieved. Interfering ions such as $F^-$, HP$O_4^{2-}$, $Fe^{2+}$, $Fe^{3+}$, $Mn^{2+}$, and other anions were tested, interference did not occur up to 1,000mg/L of ion concentration and up to 2,CO0mg/L of sulfate ion con-centration. This method was applied for the determination of aluminium ion in tap water and ground water of Jeonju and the Gochang area. The results showed that the aluminium residual in tap water of the Jeonju area was at a mean of 0.478mg/L and that in tap water of the Gochang area was at a mean of 0.278mg/L. Aluminium ion residual of the tap waters in the Jeonju area was higher level than that in the Gochang area. Aluminium residual in the ground water of the Jeonju area was 0.386 mg/L and was lower compared to 0.564 mg/L for the Gochang area.

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The Proteasome Inhibitor MG132 Sensitizes Lung Cancer Cells to TRAIL-induced Apoptosis by Inhibiting NF-κ Activation (폐암세포주에서 NFκ 활성 억제를 통한 Proteasome 억제제 MG132의 TRAIL-유도성 Apoptosis 감작 효과)

  • Seo, Pil Won;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.6
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    • pp.476-486
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    • 2008
  • Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 2. Intraeryrhrocytic culture of Babesia gibsoni by microaerophilous stationary phase(MASP) (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 2. Babesia gibsoni의 적혈구내 배양)

  • Suh, Myung-deuk;Joo, Bo-hyun
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.359-365
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    • 1998
  • This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

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Effects of Hormones on the Proliferation of Stromal Vascular Cells from Hanwoo Cattle Adipose Tissues

  • Lee, S.C.;Lee, H.J.;Kim, D.W.;Kim, J.W.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.2
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    • pp.161-166
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    • 2000
  • This study was designed to determine the effects of the insulin-like growth factor (IGF-1) and estradiol $17-{\beta}$ on the in vitro proliferation of stromal vascular cell from Hanwoo omental, subcutaneous, intermuscular and intramuscular adipose tissues. Cells were cultured in M199+20% newborn calf serum and the proliferation of cells was measured by direct microscopic cell counting and change of genomic DNA amount. Cell numbers increased slightly over the first 72 hour of culture and then increased greatly, regardless of adipose tissue depots. In IGF-1 treatment, the number of omental preadipocytes maintained highest level from the beginning to the 20th day of culture. However, in estradiol-$17{\beta}$ treatment, those tended to be lower than the control from the beginning of culture and significantly lower at the 24th day. When IGF-1 was added to subcutaneous preadipocytes, the numbers of cells were higher from 11th day than those from other treatments, although there was no statistical significance. For intermuscular preadipocytes treated with IGF-1, its numbers were significantly (p<0.05) higher at 11th day, and in the other days it showed a similar tendency to those of the subcutaneous tissue. In this experiment, preadipocytes were taken from 24 month old fully matured steers and the highest proliferation rate was shown in intramuscular tissue followed by those of subcutaneous preadipocytes. Addition of $5{\mu}M$ estradiol-$17{\beta}$ to the growth medium failed to promote the replication of Hanwoo preadipocytes, as indicated by direct cell counts and total genomic DNA content. As the culture period proceeded, the amounts of DNA were increased, but the patterns of increment were not consistent with the results of cell numbers.

Antioxidant Activity of Saururus chinensis Pretreated by Mechanochemical Technology - Nitrite Scavenging and Electron Donating Ability -

  • Song, Won-Seob;Park, Keum-Joo;Choung, Eui-Su
    • Korean Journal of Plant Resources
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    • v.24 no.3
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    • pp.314-318
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    • 2011
  • Saururus chinensis is used as a raw material of an anti-rheumatic, poultice, sedative and stomachic. The tea of dried Saururus chinensis leaves has a function of relaxing pains in back and breast and of treating stomach ailments. The plant has the functions of anti-inflammatory, depurative, diuretic, febrifuge and refrigerant. The extraction of functional material is carried out by methanol and ethanol solvents. Mechanochemical grinding is applied as a pretreatment process before extraction to enhance the extraction efficiency by increasing the surface area of the materials while changing the chemical properties of the materials. Extraction of functional materials from the Saururus chinensis after grinding as a pretreatment using the mechanochemical technology was accomplished in this study to investigate the effect of grinding on the antioxidant activities of the extract. Saururus chinensis was ground by the planetary ball mill and the morphology was analyzed by SEM. Mechanochemical pretreatment increased the yields of functional materials from 5.9 g to 6.4, 7.0, and 8.1 g after grinding of 30 minutes, 1 hour and 2 hours, respectively. Nitrite scavenging ability increased from 53.0-71.0% to 61.0-79.0% for the methanol extraction in the solution of pH 1.2. Also, Electron donating abilities were increased from 7.90-33.51% to 11.97-38.51% for the methanol extraction. The extract concentration for the half inhibition of DPPH radicals was reduced from $278{\mu}g/mL$ in the original sample to $263{\mu}g/mL$ in the sample after grinding for the methanol extraction.

Current Status(1994) of Neonatal Surgery in Korea -Survey among the members of Korean Association of Pediatric Surgeons- (1994년도 한국신생아외과의 현황 - 대한소아외과학회 회원대상 전국조사 -)

  • Kim, W.K.;Kim, S.Y.;Kim, S.K.;Kim, I.K.;Kim, J.C.;Park, K.W.;Park, Y.S.;Park, W.H.;Park, J.S.;Song, Y.T.;Oh, S.M.;Lee, D.S.;Lee, M.D.;Lee, S.C.;Chang, S.I.;Chung, E.S.;Jung, P.M.;Joo, J.S.;Choi, S.O.;Huh, Y.S.;Hwang, E.H.
    • Advances in pediatric surgery
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    • v.2 no.1
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    • pp.26-32
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    • 1996
  • To understand the current status of neonatal surgery in Korea, a survey was made among the 27 members of Korean Association of Pediatric Surgeons. Response rate among surgeons was 78 percent, eighteen hospitals participated in this study. Five hundred and three cases of neonatal surgical patients were analyzed. In Korea, about 50% of cases were treated at the hospital in the capital city area. Regional number of patients were closely related to the regional population. Imperforate anus(19%), atresia/stenosis of gut(12%), and Hirschsprung's diseases(12%) were sitting at the top in the list. Majority of operation was done within the first week of life, especially during the first 24 hour period. Eighty per cent was major or so called index cases. Mortality in so-called index cases was 17%. High mortality was observed in patients with diaphragmatic hernia(47%), gastrointestinal perforation(65%) and esophageal atresia(28%). Low birth weight babies showed higher mortality in gastrointestinal perforation, esophageal atresia and abdominal wall defect. These were compared to 1993 survey of Japanese Society of Pediatric Surgeons.

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Effects of Salinity and Moisture Content on Aerobic Composting of Food Wastes (염분도와 수분함량이 음식폐기물의 호기성 퇴비화에 미치는 영향)

  • 박석환
    • Journal of Environmental Health Sciences
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    • v.24 no.1
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    • pp.120-131
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    • 1998
  • This study was performed to define the physicochemical characteristics of food waste and food wastewater, and to find the effect of moisture content variation and salinity variation on aerobic composting for food wastes. In moisture content variation experiment, the samples of 2-1, 2-2, 2-3 and 24 were prepared by the moisture content of 83.8%, 70.9%, 64.8% and 45.1%, respectively. In salinity variation experiment, the samples of 3-1, 3-2, 3-3 and 3-4 were prepared by the salinity of 0.99%, 1. 69%, 1.75% and 2.34%, respectively. In both experiments, aerobic composting reactors were operated by the mode which was composed of half an hour's stirring and 2 hour's aeration per day, for 45 days. The followings are the conclusions that were derived from this study. 1. In the study of physicochemical characteristics of food waste and food wastewater, the values of pH were 4.19 and 3.96, the values of salinity were 0.91% and 1.17%, and the values of conductivity were 7.6 mS/cm and 18.2 mS/cm, respectively. 2. In food waste, the moisture content was 60.3%, organic compound content was 96.1%, total carbon was 48.0%, total nitrogen was 1.5%(therefore, C/N ratio was 32), and the concentration of total phosphorus was 1.34 mg/kg. 3. The time of temperature ascending was delayed, the highest temperature was lowered, the duration period of high temperature was shortened by the increasing of moisture content. In the higher moisture content, anaerobic condition was formed, bad smell was released, insects were gathered and multiplicated, and the reaction rate of composting was reduced. 4. In moisture content experiment, C/N ratios were changed from the range of 31.2-34.8 at the beginning phase to that of 20.4-28.4 at the last phase. 5. In salinity experiment, the reduction rate of volume was increased(40.3%) when the salinity was decreased(0.99%). Also, the reduction rate of mass was increased(51.8%) when the salinity was decreased(0.99%). This fact denotes that salinity hinders the process of composting. 6. the concentrations of total nitrogen and total phosphorus were increased from 0.74% to 1.10%, and from 0.82 mg/kg to 3.44 mg/kg, respectively when the salinity was decreased from 2.34% to 0.99%.

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Inflammation Scan Using $^{99m}Tc-HMPAO$ Labelled Leukocytes ($^{99m}Tc-HMPAO$를 이용한 자가백혈구표지 및 그를 이용한 염증병소의 스캔)

  • Yang, Woo-Jin;Chung, Soo-Kyo;Shinn, Kyung-Sub;Bahk, Yong-Whee;Kim, Hoon-Kyo
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.2
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    • pp.219-223
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    • 1989
  • Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

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Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.