• Title/Summary/Keyword: 기질 농도

Search Result 960, Processing Time 0.021 seconds

Effects of High Glucose and Advanced Glycosylation Endproducts(AGE) on the in vitro Permeability Model (당과 후기당화합물의 생체 외 사구체여과율 모델에 대한 역할)

  • Lee Jun-Ho;Ha Tae-Sun
    • Childhood Kidney Diseases
    • /
    • v.10 no.1
    • /
    • pp.8-17
    • /
    • 2006
  • Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.

  • PDF

Time Course Change of Phagocytes and Proinflammatory Activities in BALF in Endotoxin-induced Acute Lung Injury (시간별 내독소 정맥주입으로 유발된 급성폐손상의 변화양상에 대한 고찰)

  • Moon, Seung-Hyug;Oh, Je-Ho;Park, Sung-Woo;NamGung, Eun-Kyung;Ki, Shin-Young;Im, Gun-Il;Jung, Sung-Whan;Kim, Hyeon-Tae;Uh, Soo-Tack;Kim, Yong-Hoon;Park, Choon-Sik;Jin, Byeng-Weon
    • Tuberculosis and Respiratory Diseases
    • /
    • v.44 no.2
    • /
    • pp.360-378
    • /
    • 1997
  • Background : Severe acute lung injury(ALI), also known as the adult respiratory distress syndrome(ARDS), is a heterogenous nature of dynamic and explosive clinical synrome that exacts a mortality of approximately 50%. Endotoxin(ETX) is an abundant component of the outer membrane of gram-negative bacteria capable of inducing severe lung injury in gram-negative sepsis and gram-negative bacterial pneumonia, which are among the most common predisposing causes of ARDS. The influx of PMNs into airway tissue is a pathological hallmark of LPS-induced lung injury. And there is a substantial evidence suggesting that cytokines are important mediators of lung injury in gram-negative sepsis. However, the kinetics of phagocytes and cytokines by an exact time sequence and their respective pathogenic importance remain to be elucidated. This study was performed to investigate the role of phagocytes and proinflammatory cytokines in ETX-induced ALI through a time course of changes in the concentration of protein, $TNF{\alpha}$ and IL-6, and counts of total and its differential cells in BALF. The consecutive histologic findings were also evaluated. Method : The experimental animals, healthy male Sprague-Dawley, weighted $200{\pm}50g$, were divided into control- and ALI- group. ALI was induced by an intravenous administration of ETX, 5mg/kg. Above mentioned all parameters were examined at 0(control), 3, 6, 24, 72 h after administration of ETX. $TNF{\alpha}$ and IL-6 cone. in BALF were measured by a bioassay. Results : The protein concentration and total leukocyte count(TC) in BALF was significantly increased at 3h compared to controls(p < 0.05). The protein conc. was significantly elavated during observation period, but TC was significantly decreased at 72h(p < 0.05 vs. 24h). There was a close relationship between TC and protein cone. in BALF(r = 0.65, p < 0.001). The PMN and monocyte count was well correlated with TC in BALF, and the correlation of PMN(r = 0.97, p < 0.001) appeared to be more meaningful than that of monocyte(r = 0.61, p < 0.001). There was also a significant correlation between protein cone. and PMN or monocyte count in BALF(PMN vs. monocyte : r = 0.55, p < 0.005 vs. r = 0.64, p < 0.001). The count of monocyte was significantly elavated during observation period though a meaningful reduction of PMN count in BALF at 72h, this observation suggested that monocyte may, at least, partipate in the process of lung injury steadly. In this study, there was no relationship between IL-6 and $TNF{\alpha}$ cone., and $TNF{\alpha}$ but not IL-6 was correlated with TC(r = 0.61, p < 0.05) and monocyte(r = 0.67, p < 0.05) in BALF only at 3, 6h after ETX introduced. In particular, the IL-6 cone. increased earlier and rapidly peaked than $TNF{\alpha}$ cone. in BALF. In histologic findings, the cell counts of lung slices were increased from 3 to 72h(p < 0.001 vs. NC). Alveolar wall-thickness was increased from 6 to 24h(p < 0.001 vs. NC). There was a significant correlation between the cell counts of lung slices and alveolar wall-thickness(r= 0.61, p < 0.001). This result suggested that the cellular infiltrations might be followed by the alterations of interstitium, and the edematous change of alveolar wall might be most rapidly recovered to its normal condition in the process of repair. Conclusion : We concluded that although the role of PMN is partly certain in ETX-induced ALI, it is somewhat inadequate to its known major impact on ALL Alveolar macrophage and/or non-immune cells such as pulmonary endothelial or epithelial cells, may be more importantly contributed to the initiation and perpetual progression of ETX-induced ALI. The IL-6 in ETX-induced ALI was independent to $TNF{\alpha}$, measured by a bioassay in BALF. The early rise in IL-6 in BALF implies multiple origins of the IL-6.

  • PDF

Application of Westgard Multi-Rules for Improving Nuclear Medicine Blood Test Quality Control (핵의학 검체검사 정도관리의 개선을 위한 Westgard Multi-Rules의 적용)

  • Jung, Heung-Soo;Bae, Jin-Soo;Shin, Yong-Hwan;Kim, Ji-Young;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
    • /
    • v.16 no.1
    • /
    • pp.115-118
    • /
    • 2012
  • Purpose: The Levey-Jennings chart controlled measurement values that deviated from the tolerance value (mean ${\pm}2SD$ or ${\pm}3SD$). On the other hand, the upgraded Westgard Multi-Rules are actively recommended as a more efficient, specialized form of hospital certification in relation to Internal Quality Control. To apply Westgard Multi-Rules in quality control, credible quality control substance and target value are required. However, as physical examinations commonly use quality control substances provided within the test kit, there are many difficulties presented in the calculation of target value in relation to frequent changes in concentration value and insufficient credibility of quality control substance. This study attempts to improve the professionalism and credibility of quality control by applying Westgard Multi-Rules and calculating credible target value by using a commercialized quality control substance. Materials and Methods : This study used Immunoassay Plus Control Level 1, 2, 3 of Company B as the quality control substance of Total T3, which is the thyroid test implemented at the relevant hospital. Target value was established as the mean value of 295 cases collected for 1 month, excluding values that deviated from ${\pm}2SD$. The hospital quality control calculation program was used to enter target value. 12s, 22s, 13s, 2 of 32s, R4s, 41s, $10\bar{x}$, 7T of Westgard Multi-Rules were applied in the Total T3 experiment, which was conducted 194 times for 20 days in August. Based on the applied rules, this study classified data into random error and systemic error for analysis. Results: Quality control substances 1, 2, and 3 were each established as 84.2 ng/$dl$, 156.7 ng/$dl$, 242.4 ng/$dl$ for target values of Total T3, with the standard deviation established as 11.22 ng/$dl$, 14.52 ng/$dl$, 14.52 ng/$dl$ respectively. According to error type analysis achieved after applying Westgard Multi-Rules based on established target values, the following results were obtained for Random error, 12s was analyzed 48 times, 13s was analyzed 13 times, R4s was analyzed 6 times, for Systemic error, 22s was analyzed 10 times, 41s was analyzed 11 times, 2 of 32s was analyzed 17 times, $10\bar{x}$ was analyzed 10 times, and 7T was not applied. For uncontrollable Random error types, the entire experimental process was rechecked and greater emphasis was placed on re-testing. For controllable Systemic error types, this study searched the cause of error, recorded the relevant cause in the action form and reported the information to the Internal Quality Control committee if necessary. Conclusions : This study applied Westgard Multi-Rules by using commercialized substance as quality control substance and establishing target values. In result, precise analysis of Random error and Systemic error was achieved through the analysis of 12s, 22s, 13s, 2 of 32s, R4s, 41s, $10\bar{x}$, 7T rules. Furthermore, ideal quality control was achieved through analysis conducted on all data presented within the range of ${\pm}3SD$. In this regard, it can be said that the quality control method formed based on the systematic application of Westgard Multi-Rules is more effective than the Levey-Jennings chart and can maximize error detection.

  • PDF

Rapid Statistical Optimization of Cultural Conditions for Mass Production of Carboxymethylcellulase by a Newly Isolated Marine Bacterium, Bacillus velezensis A-68 from Rice Hulls (통계학적 방법을 사용한 해양미생물 Bacillus velezensis A-68균주의 섬유소 분해효소 생산 조건 최적화)

  • Kim, Bo-Kyung;Kim, Hye-Jin;Lee, Jin-Woo
    • Journal of Life Science
    • /
    • v.23 no.6
    • /
    • pp.757-769
    • /
    • 2013
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater, identified as Bacillus velezensis by analyses of 16S rDNA and partial sequences of the gyrA, and designated as B. velezensis A-68. The optimal conditions for production of CMCase by B. velezensis A-68 were established using response surface methodology (RSM). The optimal concentrations of rice hulls and yeast extract, and initial pH of the medium for cell growth were 60.2 g/l, 7.38 g/l, and 7.18, respectively, whereas those for production of CMCase were 50.0 g/l, 5.00 g/l, and 7.30. The analysis of variance (ANOVA) implied that the most significant factor for cell growth as well as production of CMCase was yeast extract. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ in the medium for cell growth were 7.50, 1.00, 0.10, and 0.80 g/l, respectively, which were the same as those for production of CMCase. The optimal temperatures for cell growth and production of CMCase were 30 and $35^{\circ}C$, respectively. The maximal production of CMCase under optimized conditions was 83.8 U/ml, which was 3.3 times higher than that before optimization. In this study, rice hulls, agro-byproduct, were developed as a substrate for production of CMCase and time for production of CMCase was reduced to 3 days using a newly isolated marine bacterium.

Protein Engineering of Flavin-containing Monooxygenase from Corynebacterium glutamicum for Improved Production of Indigo and Indirubin (인디고와 인디루빈의 생산을 증대하기 위한 플라빈-함유 모노옥시게나제의 단백질공학)

  • Jung, Hye Sook;Jung, Hae Bin;Kim, Hee Sook;Kim, Chang Gyeom;Lee, Jin Ho
    • Journal of Life Science
    • /
    • v.28 no.6
    • /
    • pp.656-662
    • /
    • 2018
  • Flavin-containing monooxygenases from Corynebacterium (cFMOs) were mutagenized based on homology modeling to develop variants with an enhanced indigoid production capability. The four mutants, F170Y, A210G, A210S, and T326S, which fused to a maltose-binding protein (MBP), were constructed, and their biochemical properties were characterized. Of these, purified MBP-T326S required a higher concentration of exogenous FAD (100 mM) than the wild-type MBP-cFMO for optimal activity and showed a 3.8-fold increase in the $k_{cat}/K_m$ value at $100{\mu}M$ FAD compared to that of MBP-cFMO at $2{\mu}M$ FAD. The indole oxygenase activities of MBP-T326S decreased to 63-77% compared to that of the MBP-cFMO In addition, MBP-T326S displayed a very low level of futile NADPH oxidase activities (21-24%) in the absence of a substrate. Mutant proteins except for T326S displayed similar $K_m$ and increased $k_{cat}/K_m$ values compared to the wild-type. MBP-F170Y and -A210S mutants showed elevated indole oxygenase activity higher than 3.1- and 2.9-fold, respectively, in comparison with MBP-cFMO. When indigoid production was carried out in LB broth with 2.5 g/l of tryptophan, Escherichia coli expressing cFMO produced 684 mg/l of indigo and 104 mg/l of indirubin, while cells harboring T326S produced 1,040 mg/l of indigo and 112 mg/l of indirubin. The results indicate that the production of indigo was 13% higher when compared to a previous report in which an E. coli expressing FMO from Methylophaga produced 920 mg/l of indigo. The protein engineering of cFMO based on homology modeling provided a more rational strategy for developing indigoid-producing strains.

Studies on the Enzymes Produced by Basidiomycetes -Part II. Properties of Cellulase and Xylanase- (담자균(擔子菌)이 생산(生産)하는 효소(酵素)에 관(關)한 연구(硏究) -제2보(第二報) Cellulase 및 Xylanase의 성질(性質)-)

  • Hong, Jai-Sik;Kwon, Yong-Ju
    • Applied Biological Chemistry
    • /
    • v.24 no.4
    • /
    • pp.260-266
    • /
    • 1981
  • Some properties of cellulase and xylanase produced from Pleurotus ostreatus 301 and Lentinus edodes 3-1 during its growth in rice straw medium were investigated. The cellulase activities of P. ostreatus 301 and L. edodes 3-1 were increased in proportion to substrate concentration within 0.6% and 0.8%, respectively, and xylanase activities of two strains were increased within 1%. The reducing sugar production of cellulase and xylanase in two strains were proportionaly increased until 30 min. and 60 min. respectively. The opium pH for cellulase activities of P. ostreatus 301 and L. edodes 3-1 were pH 4.0 and pH 4.5, respectively, and xylanase activities of two strains were pH 5.0. The stable pH range for cellulase activities of P. ostreatus 301 was within 4.0 to 6.0 and L. edodes 3-1 was within 3.0 to 5.0, Xylanase activities of P. ostreatus 301 was within 4.5 to 6.0 and L. edodes 3-1 was within 3.5 to 6.0. The optium temperature for cellulase activities of P. ostraeatus 301 and L. edodes 3-1 were $40^{\circ}C$ and $50^{\circ}C$, respectively, but xylanase activities of P. ostreatus 301 and L. edodes 3-1 were $50^{\circ}C$ and $45^{\circ}C$, respectively. Thermal stability of enzymes were below of optimum temperature and these were mostly inactivate at $70^{\circ}C$ for 10 min of the metalic ions tested, cellulase activities of L. edodes 3-1 was increased by $Co^{++},\; Mg^{++}$ at the concentration of $10^{-2}M$, but were greatly inhibited by $Hg^{++},\;Cu^{++}$ in two strains. Xylanase activities were increased by $Ca^{++},\;Co^{++},\;Mg^{++}$ and $Cd^{++}$ but was greatly inhibited by $Hg^{++}$.

  • PDF

Evaluation of Solidified Fuel Value of Dairy Cattle Manure Digested by Semi-Dry Anaerobic Digestion Method (젖소분뇨 반 건식 혐기소화 잔재물의 고체연료화 가능성 평가)

  • Jeong, Kwang-Hwa;Kim, Jung Kon;Lee, Dong-jun;Cho, Won-Mo;Ravindran, B.;Kwag, Jung-Hoon
    • Journal of the Korea Organic Resources Recycling Association
    • /
    • v.24 no.4
    • /
    • pp.95-103
    • /
    • 2016
  • The objective of this study was to investigate feasibility of semi-dry anaerobic digestion using dairy cattle manure and to evaluate solidified fuel value of semi-dry anaerobic digestate. To evaluate semi-dry anaerobic digestion using dairy cattle manure, 950 mL bottle type anaerobic reactor was set in the constant temperature room maintained at $35^{\circ}C$. To produce anaerobic digestate for making solidified fuel, acrylic cylindrical anaerobic digester(1,000 mm width ${\times}$ 450 mm height) was set in the constant room temperature to carry out batch test of semi-dry anaerobic digestion using same dairy cattle manure. Moisture content of dairy cattle manure and inoculum solution for anaerobic digestion were 80.64% and 96.83%, respectively. The dairy cattle manure and the inoculum solution was mixed by 1:1 ratio(v/v) for anaerobic digestion. Water content and VS/TS(Volatile Solids/Total Solids) of mixture of substrate and inoculum were 89.74% and 83.35%, respectively. In case of non-inoculated anaerobic digester, the biogas was not produced. By the semi-dry anaerobic digestion, the calorific value of the digestate was reduced by 20% compare to fresh dairy cattle manure. In other hand, ash content increased from 15% to 18.4%. The contents of Cr, Pb, Cd and S of pellet produced from anaerobically digested dairy cattle manure were not against the standard regulation for livestock manure solidified fuel. Therefore, it can be used as fuel that anaerobic digestate produced after semi-dry anaerobic digestion using dairy cattle manure.

Effects of Dry Eye Symptoms on Work Productivity and General Activity in Newly Building (신축건물에서 안구건조증이 작업생산성과 일상활동에 미치는 영향)

  • Kim, Hyojin;Park, Chan-Jung;Lim, Byung-Seo;Kim, Ho-Hyun
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.19 no.3
    • /
    • pp.389-396
    • /
    • 2014
  • Purpose: The study examined dry eye symptoms of occupants in a newly constructed building and its effects on their work productivity and general activity. Methods: The study subjects were 33 office workers who spent more than eight hours per day on average in a new building constructed in the past three months. The indoor air quality of the new building was evaluated by measuring aldehydes, temperature and humidity. The level of dry eye symptoms was classified into normal, mild, moderate and severe by using the Ocular Surface Disease Index (OSDI). The experience of LASIK surgery, use of eye makeup, daily use time of a computer and smart-phone, and average daily working hours were also examined. The Work Limitation Productivity Questionnaire was used as the questionnaire about work productivity and general activity to measure the impairment level on a ten-point scale. Results: The concentration variation of formaldehyde in the office was $42.42{\pm}6.30{\mu}g/m^3$. The temperature and humidity were $26.2{\pm}0.70^{\circ}C$ and $40{\pm}1%$, respectively. The respondents with normal, mild, moderate and severe dry eye symptoms were 15.2%, 18.2%, 18.2% and 48.5%, respectively. The severity of dry eye symptoms and impairment of work productivity and general activity demonstrated high correlations of 0.599 and 0.655, respectively (p<0.001). Compared to the normal case, severe dry eye symptoms demonstrated significantly high impairment of work productivity and interruption of general activity (p<0.001). The case of serious dry symptoms showed the possibilities of having impairment level of work productivity and interruption of general activity above three points 3.26 times (p=0.032) and 2.25 times (p=0.045), respectively, higher than that of the normal case. Conclusions: It was confirmed that dry eye symptoms among office workers in a newly constructed building affects work productivity and general activity.

CHEMICAL DEGRADATION AND WEAR OF LIGHT-CURED COMPOSITE RESINS (광중합형 복합레진의 화학적 분해와 마모에 관한 연구)

  • Yang, Kyu-Ho;Jung, Hee-Kyung;Choi, Nam-Ki;Kim, Seon-Mi
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.34 no.2
    • /
    • pp.273-284
    • /
    • 2007
  • The aim of this study was to evaluate the resistance to degradation and to compare the wear resistance characteristics of four esthetic restorative materials in an alkaline solution(0.1N NaOH). The composite resins studied were Composan LCM flow(Promedica, Germany). Clearfil ST(Kuraray medical, Japan), Durafi VS1(Heraeus Kulzer, U.S.A), Point 4(Kerr, U.S.A). The results were as follows : 1. The mass loss of each brand was $1.02{\sim}6.04%$ and highest value in Durafil VS$(6.04{\pm}0.29%)$. 2. The sequence of the degree of degradation layer depth was in descending order by Durafil VS, Clearfil ST, Point 4 and Composan LCM flow. There were significant differences between Point 4, Composan LCM flow and the others (p<0.001). 3. The sequence of the Si loss was in descending order by Clearfil ST, Durafil VS, Composan LCM flow and Point 4. There were significant differences among the materials (p<0.001). 4. On SEM, destruction of bonding between matrix and filler and on CLSM, the depth of degradation layer of specimen surface was observed. 5. The sequence of maximum wear depth was in descending order by Durafil VS, Composan LCM flow, Point 4 and Clearfil ST. There were no significant differences among the materials (p>0.001) 6. The correlation coefficient between Si loss and degradation layer depth (r=0.892, p<0.01) and Si loss and mass loss(r=0.736, p<0.01) were relatively high. These results indicate that hydrolytic degradation, wear and another factor may consider as evaluation factors of composite resins.

  • PDF

Distribution and Growth of Bacteria in the Hypertrophic Lake Shiwha (과영양성 시화호에서 박테리아의 분포 및 성장)

  • Choi, Dong-Han;Kang, Sulk-Won;Song, Ki-Don;Huh, Sung-Hoi;Cho, Byung-Cheol
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
    • /
    • v.2 no.2
    • /
    • pp.92-100
    • /
    • 1997
  • Distribution of bacterial abundance and production was investigated during October, 1995-August, 1996 in Lake Shiwha constructed artificially in 1994. Its water column was distinguished by two layers: the brackish surface layer with salinity ranged from 6 to 20‰ and the saline hypoxic/anoxic bottom layer with salinity of 17 to 27‰ Except for samples collected in March, 1996 (on average 13 ${\mu}g\;l^{-1}$), chlorophyll a concentration ranged from 27.6 to 249.5 ${\mu}g\;l^{-1}$ in the euphotic zone, indicating the hypertrophic condition of Lake Shiwha during most of the studied period. In this study, bacterial productions measured by $^3H$-thymidine incorporation method were similar to those by $^{14}C$-leucine incorporation method. In hypertrophic, surface waters of Lake Shiwha, bacterial abundance and production ranged from 1.4 to $19.5{\times}10^9\;cells\;l^{-1}$ and from 1.6 to $126.5{\times}10^7\;cells\;l^{-1}\;h^{-1}$ respectively; 2 to 4 fold and 2 to 30 fold higher than those in eutrophic coastal waters outside of Lake Shiwha, respectively. Turnover times of bacterial community in the surface layer of Lake Shiwha ranged from 0.2 to 8.9 day, indicating that bacteria in the lake seemed to adapt to the hypertrophic condition. In the hypoxic bottom layer, bacterial abundance and production was up to 3 fold and 20 fold lower than those in the surface layer, and showed slow bacterial growth. Significant correlations between the bacterial abundance, production, and community turnover time with water temperature indicate water temperature was the important factor controlling distribution and growth of bacteria. However, during summer season, bacterial production seemed to be regulated by supply of substrates.

  • PDF