• Title/Summary/Keyword: reduction of pressure

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Inflammatory Reponse of the Lung to Hypothermia and Fluid Therapy after Hemorrhagic Shock in Rats (흰쥐에서 출혈성 쇼크 후 회복 시 저체온법 및 수액 치료에 따른 폐장의 염증성 변화)

  • Jang, Won-Chae;Beom, Min-Sun;Jeong, In-Seok;Hong, Young-Ju;Oh, Bong-Suk
    • Journal of Chest Surgery
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    • v.39 no.12 s.269
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    • pp.879-890
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    • 2006
  • Background: The dysfunction of multiple organs is found to be caused by reactive oxygen species as a major modulator of microvascular injury after hemorrhagic shock. Hemorrhagic shock, one of many causes inducing acute lung injury, is associated with increase in alveolocapillary permeability and characterized by edema, neutrophil infiltration, and hemorrhage in the interstitial and alveolar space. Aggressive and rapid fluid resuscitation potentially might increased the risk of pulmonary dysfunction by the interstitial edema. Therefore, in order to improve the pulmonary dysfunction induced by hemorrhagic shock, the present study was attempted to investigate how to reduce the inflammatory responses and edema in lung. Material and Method: Male Sprague-Dawley rats, weight 300 to 350 gm were anesthetized with ketamine(7 mg/kg) intramuscular Hemorrhagic Shock(HS) was induced by withdrawal of 3 mL/100 g over 10 min. through right jugular vein. Mean arterial pressure was then maintained at $35{\sim}40$ mmHg by further blood withdrawal. At 60 min. after HS, the shed blood and Ringer's solution or 5% albumin was infused to restore mean carotid arterial pressure over 80 mmHg. Rats were divided into three groups according to rectal temperature level($37^{\circ}C$[normothermia] vs $33^{\circ}C$[mild hypothermia]) and resuscitation fluid(lactate Ringer's solution vs 5% albumin solution). Group I consisted of rats with the normothermia and lactate Ringer's solution infusion. Group II consisted of rats with the systemic hypothermia and lactate Ringer's solution infusion. Group III consisted of rats with the systemic hypothermia and 5% albumin solution infusion. Hemodynamic parameters(heart rate, mean carotid arterial pressure), metabolism, and pulmonary tissue damage were observed for 4 hours. Result: In all experimental groups including 6 rats in group I, totally 26 rats were alive in 3rd stage. However, bleeding volume of group I in first stage was $3.2{\pm}0.5$ mL/100 g less than those of group II($3.9{\pm}0.8$ mL/100 g) and group III($4.1{\pm}0.7$ mL/100 g). Fluid volume infused in 2nd stage was $28.6{\pm}6.0$ mL(group I), $20.6{\pm}4.0$ mL(group II) and $14.7{\pm}2.7$ mL(group III), retrospectively in which there was statistically a significance between all groups(p<0.05). Plasma potassium level was markedly elevated in comparison with other groups(II and III), whereas glucose level was obviously reduced in 2nd stage of group I. Level of interleukine-8 in group I was obviously higher than that of group II or III(p<0.05). They were $1.834{\pm}437$ pg/mL(group I), $1,006{\pm}532$ pg/mL(group II), and $764{\pm}302$ pg/mL(group III), retrospectively. In histologic score, the score of group III($1.6{\pm}0.6$) was significantly lower than that of group I($2.8{\pm}1.2$)(p<0.05). Conclusion: In pressure-controlled hemorrhagic shock model, it is suggested that hypothermia might inhibit the direct damage of ischemic tissue through reduction of basic metabolic rate in shock state compared to normothermia. It seems that hypothermia should be benefit to recovery pulmonary function by reducing replaced fluid volume, inhibiting anti-inflammatory agent(IL-8) and leukocyte infiltration in state of ischemia-reperfusion injury. However, if is considered that other changes in pulmonary damage and inflammatory responses might induce by not only kinds of fluid solutions but also hypothermia, and that the detailed evaluation should be study.

Use of Noninvasive Mechanical Ventilation in Acute Hypercapnic versus Hypoxic Respiratory Failure (급성 환기부전과 산소화부전에서 비침습적 환기법의 비교)

  • Lee, Sung Soon;Lim, Chae-Man;Kim, Baek-Nam;Koh, Younsuck;Park, Pyung Hwan;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.987-996
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    • 1996
  • Background : We prospectively evaluated the applicability and effect of noninvasive ventilation (NIV) in acute respiratory failure and tried to find out the parameters that could predict successful application of NIV. Methods : Twenty-six out of 106 patients with either acute ventilatory failure (VF: $PaCO_2$ > 43 mm Hg with pH < 7.35) or oxygenation failure (OF: $PaO_2/AO_2$ < 300 mm Hg with $pH{\geq}7.35$) requiring mechanical ventilation were managed by NIV (CPAP + pressure suppon, or BiPAP) with face mask. Eleven out of 19 cases with VF (57.9%) (M : F=7 : $55.4{\pm}14.6$ yrs) and 15 out of 87 cases with OF (17.2%) (M : F=12 : 3, $50.6{\pm}15.6$ yrs) were s uilable for NIY. Respiratory rates, arterial blood gases and success rate of NIV were analyzed in each group. Results: 81.8% (9/11) of YF and 40% (6/15) of OF were successfully managed on NIV and were weruled from mechanical ventilator without resorting to endotracheal intubation. Complications were noted in 2 cases (nasal skin necrosis 1, gaseous gastric distension 1). In NIV for ventilatory failure, the respiration rate was significantly decreased at 12 hour of NIV ($34{\pm}9$ /min pre-NIV, $26{\pm}6$ /min at 12 hour of NIV, p=0.045), while $PaCO_2$ ($87.3{\pm}20.6$ mm Hg pre-NIV, $81.2{\pm}9.1$ mm Hg at 24 hour of NIV) and pH ($7.26{\pm}0.04$, $7.32{\pm}0.02$, respectively, p <0.05) were both significantly decreased at 24 hour of NIV In NIV for oxygenation failure, $PaCO_2$ were not different between the successful and the failed cases at pre-NIV and till 12 hours after NIV. The $PaO_2/FIO_2$ ratio, however, significantly improved at 0.5 hour of NIV in successful cases and were maintained at around 200 mm Hg (n=6 : at baseline, 0.5h, 6h, 12h : $120.0{\pm}19.6$, $218.9{\pm}98.3$, $191.3{\pm}55.2$, $232.8{\pm}17.6$ mm Hg, respectively, p=0.0211), but it did not rise in the failed cases (n=9 : $127.9{\pm}63.0$, $116.8{\pm}24.4$, $100.6{\pm}34.6$, $129.8{\pm}50.3$ mm Hg, respectively, p=0.5319). Conclusion : From the above results we conclude that NIV is effective for hypercapnic respiratory failure and its success was heralded by reduction of respiration rale before the reduction in $PaCO_2$ level. In hypoxic respiratory failure, NIV is much less effective, and the immediate improvement of $PaO_2/FIO_2$ ratio at 0.5h after application is thought to be a predictor of successful NIV.

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One-stop Evaluation Protocol of Ischemic Heart Disease: Myocardial Fusion PET Study (허혈성 심장 질환의 One-stop Evaluation Protocol: Myocardial Fusion PET Study)

  • Kim, Kyong-Mok;Lee, Byung-Wook;Lee, Dong-Wook;Kim, Jeong-Su;Jang, Yeong-Do;Bang, Chan-Seok;Baek, Jong-Hun;Lee, In-Su
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.33-37
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    • 2010
  • Purpose: In the early stage of using PET/CT, it was used to damper revision but recently shows that CT with MDCT is commonly used and works well for an anatomical diagnosis. This hospital makes the accuracy and convenience more higher in the diagnosis and evaluate of coronary heart disease through concurrently running myocardial perfusion SPECT examination, myocardial PET examination with FDG, and CT coronary artery CT angiography(coronary CTA) used PET/CT with 64-slice. This report shows protocol and image based on results from about 400 coronary heart disease examinations since having 64 channels PET/CT in July 2007. Materials and Methods: An Equipment for this examination is 64-slice CT and Discovery VCT (DVCT) that is consisted of PET with BGO ($Bi_4Ge_3O_{12}$) scintillation crystal by GE health care. First myocardial perfusion SPECT with pharmacologic stress test to reduce waiting time of a patient and get a quick diagnosis and evaluation, and right after it, myocardial FDG PET examination and coronary CTA run without a break. One-stop evaluation protocol of ischemic heart disease is as follows. 1)Myocardial perfusion SPECT with pharmacologic stress: A patient is injected with $^{99m}Tc$-MIBI 10 mCi and does not have any fatty food for myocardial PET examination and drink natural water with ursodeoxcholic acid 100 mg and we get SPECT image in an hour. 2)Myocardial FDG PET: To reduce blood fatty content and to increase uptake of FDG, we used creative oral glucose load using insulin and Acipimox to according to blood acid content. A patient is injected with $^{18}F$-FDG 5 mCi for reduction of his radiation exposure and we get a gated image an hour later and get delay image when we need. 3) Coronary CTA: The most important point is to control heart rate and to get cooperation of patient's breath. In order to reduce a heart rate of him or her below 65 beats, let him or her take beta blocker 50 mg ~ 200 mg after a consultation with a doctor about it and have breath-practices then have the examination. Right before the examination, we spray isosorbide dinitrate 3 to 5 times to lower tension of bessel wall and to extension a blood wall of a patient. It makes to get better the shape of an anatomy. At filming, a patient is injected CT contrast with high pressure and have enough practices before the examination in order to have no problem. For reduction of his radiation exposure, we have to do ECG-triggered X-ray tube modulation exposure. Results: We evaluate coronary artery stenosis through coronary CTA and study correlation (culprit vessel check) of a decline between stenosis and perfusion from the myocardial perfusion SPECT with pharmacologic stress, coronary CTA, and can check viability of infarction or hibernating myocardium by FDG PET. Conclusion: The examination makes us to set up a direction of remedy (drug treatment, PCI, CABG) because we can estimate of effect from remedy, lesion site and severity. In addition, we have an advantage that it takes just 3 hours and one-stop in that all of process of examinations run in succession and at the same time. Therefore it shows that the method is useful in one stop evaluation of ischemic heart disease.

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Weight loss effects of Bariatric Surgery after nutrition education in extremely obese patients (고도비만환자에서 베리아트릭 수술 (Bariatric Surgery) 후 영양교육이 체중감량에 미치는 효과)

  • Jeong, Eun-Ha;Lee, Hong-Chan;Yim, Jung-Eun
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.30-45
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    • 2015
  • Purpose: This study was planned to determine the characteristics of extremely obese patients during Bariatric surgery and to evaluate how the difference in the number of postsurgical personal nutritional educations they received affected the weight loss. Methods: This is a retrospective study on the basis of the medical records of extremely obese patients for 15 months after receiving gastric banding. A total of 60 people were selected as the study subjects and they were divided into the Less Educated Group and the More Educated Group according to the average number of personal nutritional educations they received. We investigated both groups to determine the general characteristic, health related lifestyle habits, obesity related complications and symptoms in possession, and eating habits before their surgery, the body composition measurement result, obesity determination indices at 1, 3, 6, 9, 12, and 15 months before and after their surgery, and the biochemical parameters at 6 months before and after their surgery. Results: Body fat and weight showed rapid reduction until 6 months after the surgery, but thereafter reduced slowly depending on the result of body composition measurement. Regarding body fat and weight, the More Educated Group, who received nutrition education more often, showed significantly lower levels than the Less Educated Group at 15 months after surgery. Regarding BMI and degree of obesity, the More Educated Group showed significantly lower levels than the Less Educated Group at 15 months after surgery. Here, we were assured that BMI is reversely proportional to the number of personal nutritional educations at 15 months, which is more outstanding after surgery than before surgery. Conclusion: Long-term nutritional education is a key factor for the extremely obese patient in maintaining the effects of Bariatric surgery on weight and body fat reduction onwards. In the next stage, considering the characteristics of the study subjects, adoption of individual nutrition education is recommended for postsurgical prospective arbitration of obesity in order to monitor blood pressure, obesity related complications, symptoms in possession, and how eating habits and health related life habits change, and to judge the actual effect of the nutritional education method at the same time.

A Study on the Leakage Protection with Polypropylene Mat in Irrigation Canal (Polypropylene Mat에 의(依)한 용수로(用水路)의 누수방지(漏水防止)에 관(關)한 연구(硏究))

  • Kang, Sin-Up;Kang, Yea-Mook;Cho, Seung-Seup
    • Korean Journal of Agricultural Science
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    • v.6 no.2
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    • pp.166-184
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    • 1979
  • In order to prevent the water loss in the irrigation canal constructed on the sandy gravel layer or on the other highly permeable ground layer, lining has been practiced. Many studies have been done so far on the lining method to prevent the water loss in the irrigation canal and recently studies on the lining with plastic film or polyethylene film were also reported. However, the plastic film or polyethylene film has low strength and is liable to break, and water loss from pin hole caused by contacting with sand or gravel is highly predicted. This study was then conducted to find proper lining and buring method in canal construction of polypropylene mat after coated with vinyl, as one way to overcome the shortcoming frequently observed when plastic or usual polyehtylene film were used. Eventhough rather longer periods of experiments are needed to attain reliable and accurate results on the variation of durability, the durability of asphalt coated area, or on the damage due to freeze after burial or exposure of polypropylene mat, the experiemental results obtained during one year of period are summarized as follows: 1. The curvature at the area between canal bottom and side slope had increased stability and saved consruction cost. The relationship among the variation of curvature, the reduction of polypropylene mat and the reduced amount of soil cutting at each side slope was presented in Fig. 7 through 9. 2. The depth of covering material to protect polypropylene mat was desired to be over 30cm, considering the water depth, side slope, canal cleaning practices, traffic, or back pressure of irrigation period. 3. In order to increase the canal stability and to prevent slope erosion, sandy soil was required, to be placed under ground, and coarse gravel should cover the surface area of canal. 4. The studies on the stability of side slope in the canal should consider the passive area on the bottom and the slope should be about 1 to 2, considering the slope stability, allowable velocity and tractive force. 5. When compared with earth lining, the lining with polypropylene mat coated with vinyl was responsible to save 28% and 37% of canal lining cost, when the soil carrying distances were 500 and 700m. respectively. 6. The water interception was almost completely attained when the polypropylene mat coated with vinyl was used for lining. But further studies were assumed to be necessary for the use of asphalt since the strength of polypropylene mat connected with asphalt will vary with duration.

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The Respiratory and Hemodynamic Effects of Prone Position According to the Level of PEEP in a Dog Acute Lung Injury Model (잡종견 급성폐손상 모델에서 Prone position 시행시 PEEP 수준에 따른 호흡 및 혈류역학적 효과)

  • Lim, Chae-Man;Chin, Jae-Yong;Koh, Youn-Suck;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.140-152
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    • 1998
  • Background: Prone position improves oxygenation in patients with ARDS probably by reducing shunt Reduction of shunt in prone position is thought to be effected by lowering of the critical opening pressure (COP) of the dorsal lung because the pleural pressure becomes less positive in prone position compared to supine position. It can then be assumed that prone position would bring about greater improvement in oxygenation when PEEP applied in supine position is just beneath COP than when PEEP is above COP. Hemodynamically, prone position is expected to attenuate the lifting of cardiac fossa induced by PEEP. Based on these backgrounds, we investigated whether the effect of prone position on oxygenation differs in magnitude according to the level of PEEP applied in supine position, and whether impaired cardiac output in supine position by PEEP can be restored in prone position. Methods: In seven mongrel dogs, $PaO_2/F_1O_2$(P/F) was measured in supine position and at prone position 30 min. Cardiac output (CO), stroke volume (SV), pulse rate (PR), and pulmonary artery occlusion pressure (PAOP) were measured in supine position, at prone position 5 min, and at prone position 30 min. After ARDS was established with warmed saline lavage(P/F ratio $134{\pm}72$ mm Hg), inflection point was measured by constant flow method($6.6{\pm}1.4cm$ $H_2O$), and the above variables were measured in supine and prone positions under the application of Low PEEP($5.0{\pm}1.2cm$ $H_2O$), and Optimal PEEP($9.0{\pm}1.2cm$ $H_2O$)(2 cm $H_2O$ below and above the inflection point, respectively) consecutively. Results : P/F ratio in supine position was $195{\pm}112$ mm Hg at Low PEEP and $466{\pm}63$ mm Hg at Optimal PEEP(p=0.003). Net increase of P/F ratio at prone position 30 min, however, was far greater at Low PEEP($205{\pm}90$ mm Hg) than at Optimal PEEP($33{\pm}33$ mm Hg)(p=0.009). Compared to CO in supine position at Optimal PEEP($2.4{\pm}0.5$ L/min), CO in prone improved to $3.4{\pm}0.6$ L/min at prone position 5 min (p=0.0180) and $3.6{\pm}0.7$ L/min at prone position 30 min (p=0.0180). Improvement in CO was attributable to the increase in SV: $14{\pm}2$ ml in supine position, $20{\pm}2$ ml at prone position 5 min (p=0.0180), and $21{\pm}2$ ml at prone position 30 min (p=0.0180), but not to change in PR or PAOP. When the dogs were turned to supine position again, MAP ($92{\pm}23$ mm Hg, p=0.009), CO ($2.4{\pm}0.5$ L/min, p=0.0277) and SV ($14{\pm}1$ ml, p=0.0277) were all decreased compared to prone position 30 min. Conclusion: Prone position in a dog with saline-lavaged acute lung injury appeared to augment the effect of relatively low PEEP on oxygenation, and also attenuate the adverse hemodynamic effect of relatively high PEEP. These findings suggest that a PEEP lower than Optimal PEEP can be adopted in prone position to achieve the goal of alveolar recruitment in ARDS avoiding the hemodynamic complications of a higher PEEP at the same time.

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Recent Progress in Air-Conditioning and Refrigeration Research : A Review of Papers Published in the Korean Journal of Air-Conditioning and Refrigeration Engineering in 2016 (설비공학 분야의 최근 연구 동향 : 2016년 학회지 논문에 대한 종합적 고찰)

  • Lee, Dae-Young;Kim, Sa Ryang;Kim, Hyun-Jung;Kim, Dong-Seon;Park, Jun-Seok;Ihm, Pyeong Chan
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.29 no.6
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    • pp.327-340
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    • 2017
  • This article reviews the papers published in the Korean Journal of Air-Conditioning and Refrigeration Engineering during 2016. It is intended to understand the status of current research in the areas of heating, cooling, ventilation, sanitation, and indoor environments of buildings and plant facilities. Conclusions are as follows. (1) The research works on the thermal and fluid engineering have been reviewed as groups of flow, heat and mass transfer, the reduction of pollutant exhaust gas, cooling and heating, the renewable energy system and the flow around buildings. CFD schemes were used more for all research areas. (2) Research works on heat transfer area have been reviewed in the categories of heat transfer characteristics, pool boiling and condensing heat transfer and industrial heat exchangers. Researches on heat transfer characteristics included the results of the long-term performance variation of the plate-type enthalpy exchange element made of paper, design optimization of an extruded-type cooling structure for reducing the weight of LED street lights, and hot plate welding of thermoplastic elastomer packing. In the area of pool boiling and condensing, the heat transfer characteristics of a finned-tube heat exchanger in a PCM (phase change material) thermal energy storage system, influence of flow boiling heat transfer on fouling phenomenon in nanofluids, and PCM at the simultaneous charging and discharging condition were studied. In the area of industrial heat exchangers, one-dimensional flow network model and porous-media model, and R245fa in a plate-shell heat exchanger were studied. (3) Various studies were published in the categories of refrigeration cycle, alternative refrigeration/energy system, system control. In the refrigeration cycle category, subjects include mobile cold storage heat exchanger, compressor reliability, indirect refrigeration system with $CO_2$ as secondary fluid, heat pump for fuel-cell vehicle, heat recovery from hybrid drier and heat exchangers with two-port and flat tubes. In the alternative refrigeration/energy system category, subjects include membrane module for dehumidification refrigeration, desiccant-assisted low-temperature drying, regenerative evaporative cooler and ejector-assisted multi-stage evaporation. In the system control category, subjects include multi-refrigeration system control, emergency cooling of data center and variable-speed compressor control. (4) In building mechanical system research fields, fifteenth studies were reported for achieving effective design of the mechanical systems, and also for maximizing the energy efficiency of buildings. The topics of the studies included energy performance, HVAC system, ventilation, renewable energies, etc. Proposed designs, performance tests using numerical methods and experiments provide useful information and key data which could be help for improving the energy efficiency of the buildings. (5) The field of architectural environment was mostly focused on indoor environment and building energy. The main researches of indoor environment were related to the analyses of indoor thermal environments controlled by portable cooler, the effects of outdoor wind pressure in airflow at high-rise buildings, window air tightness related to the filling piece shapes, stack effect in core type's office building and the development of a movable drawer-type light shelf with adjustable depth of the reflector. The subjects of building energy were worked on the energy consumption analysis in office building, the prediction of exit air temperature of horizontal geothermal heat exchanger, LS-SVM based modeling of hot water supply load for district heating system, the energy saving effect of ERV system using night purge control method and the effect of strengthened insulation level to the building heating and cooling load.

The Effects of Aminophylline on the Superoxide Anion Generation of Neutrophils from Established Human Sepsis Caused by Acute Pneumonia (급성 폐렴에 의한 패혈증 환자에서 Aminophylline이 혈중 호중구의 과산화물 음이온 유리에 미치는 영향)

  • Kim, Yong-Hoon;Park, Jun-Young;Cha, Mi-Kyong;Lee, Sang-Moo;Kim, Hyeon-Tae;Uh, Soo-Taek;Chung, Yeon-Tae;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.1
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    • pp.16-22
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    • 1993
  • Background: The Microbicidal and cytotoxic activities of neutrophils are to a large extent dependent on a burst of oxidative metabolism which generates superoxide anion, hydrogen peroxide, and other reactive products of oxygen. The respiratory burst of PMN is initiated by intracellular calcium mobilization that follows immune or particular stimulation and is very sensitive to modulation by c-AMP or adenosine. Despite its antagonism against adenosine, earlier study has demonstrated potent theophylline inhibition of the PMN respiratory burst at variable ranges of blood concentrations of theophylline in the healthy normal volunteers and in the septic animals pretreated or early post-treated with aminophylline (AMPH) or pentoxifylline. However it is unclear whether theophylline inhibits the superoxide generation or not in the established human sepsis caused by acute pneumonia, as taking into consideration of the fact that full activation of neutrophils have occurred within minutes after the septic insult in the animal experiments. Methods: We measured the $O_2$ generation of peripheral arterial neutrophils obtained from 11 human septic subjects caused by acute pneumonia before and 1 hour after completion of continuous AMPH infusion. Patients were identified and studied within 48 hour of admission. All subjects were administered an intravenous loading and maintenance dose of AMPH. The generation of $O_2$ was measured at a discrete time point (60 min) by the reduction of ferricytochrome c.PMA (10 ${\mu}g/ml$) was used as a stimulating agent. PMNs were isolated at a concentration of $2{\times}10^6$ cells/ml. The arterial oxygen tension, blood pressure and heart rates were also checked to evaluate the systemic effects of AMPH in the acute pneumonia. Results: The mean serum concentration of AMPH at 60 minutes was $8.8{\pm}0.6{\mu}g/ml$. Sixty minutes after AMPH infusion the generatition of $O_2$ was decreased from $0.076{\pm}0.034$ to $0.013{\pm}0.004$(OD) (p<0.05) and from $0.177{\pm}0.044$ to $0.095{\pm}0.042$(OD) (p<0.01) in the resting and stimulated PMNs respectively. $PaO_2$ was not changed after AMPH infusion. Conclusion: AMPH may compromise host defense by significant inhibition of neutrophil release of superoxide anion and it had no effect on improving $PaO_2$ in the acute pneumonia.

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Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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Development of Near-Isogenic Line of japonica Rice Cultivar Saenuri without Lipoxygenase-3 (새누리 벼 품종 배경 lipoxygenase-3 결핍 자포니카 근동질계통 개발)

  • Park, Hyun-Su;Lee, Keon-Mi;Kim, Ki-Young;Kim, Jeong-Ju;Shin, Woon-Cheol;Baek, Man-Kee;Kim, Choon-Song;Park, Seul-Gi;Lee, Chang-Min;Suh, Jung-Pil;Cho, Young-Chan
    • Korean Journal of Breeding Science
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    • v.51 no.3
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    • pp.190-200
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    • 2019
  • It is reported that the absence of lipoxygenase-3 (LOX-3) may contribute to a reduction in stale flavor after the storage of rice. To improve the quality of stored rice of the Korean japonica rice cultivar, we conducted a breeding program to develop near-isogenic rice without LOX-3 in the genetic background of Saenuri, a mega variety of Korea. In the first step of the breeding program, we used a donor parent of LOX-3 null, Daw Dam, and a recurrent japonica parent, Sindongjin, to develop HR27873-AC12 by backcross (BC1), color test for introgression of lox-3, and anther culture for rapid fixation. In the second step, we used the donor parent, HR27873-AC12, and the recurrent parent, Saenuri, to develop HR28896-31-3-1-1 by backcross (BC1), marker-assisted selection (MAS) for lox-3, and phenotypic selection (PS) for agronomic traits. Finally, in the third step, we developed HR30960-186-2-1-2-1 (Jeonju624), derived from a cross between Saenuri and HR28896-31-3-1-1, by MAS for lox-3 and PS with high selection pressure for agronomic characteristics. Jeonju624 was confirmed with the introgression of lox-3 by molecular marker. Jeonju624 was a mid-late maturing rice with similar agronomic characteristics to Saenuri, lodging tolerance with short culm, erect plant architecture, and resistance to bacterial blight and rice stripe virus. The yield components of Jeonju624 were mostly similar to Saenuri, except for the 1,000-grain weight of brown rice. The appearance of the grain of Jeonju624 was better than that of Saenuri, and the characteristics of cooked rice were similar to those of Saenuri. In the genetic background analysis using 406 KASP (Kompetitive Allele-Specific PCR) markers, Jeonju624 was confirmed to be the near-isogenic line (NIL) of Saenuri with a 95.8% recovery rate. Jeonju624 is the NIL of Saenuri without LOX-3, and overcomes the linkage drag of Daw Dam with similar agronomic characteristics and genetic background to Saenuri. Jeonju624 can be utilized as a practical cultivar to improve the quality of stored rice, breeding material for the introgression of lox-3, and genetic material to elucidate the effect of introgressed genes.