• Toxicological Research
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• v.4 no.2
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• pp.181-188
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• 1988

This study was carried out to investigate whether the use of the high endotoxin-sensitive or the low endotoxin-sensitive rabbits (New Zealand White strain) are more suited for the pyrogen test regulated by the Korean Pharmacopeia and United State Pharmacopeia. The results obtained were as follows. 1. The ferbrile responese were increased according to increased endotoxin doses used (i.e., 0.1, 1, 5, 10ng/kg) and the febrile responses reached to its maximum between 90 mins and 120 mins after the intravenous injection of endotoxin.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.162.1-162.1
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• 2003

Endotoxin has been detected by the Limulus amoebocyte lysate (LAL) test. However, aluminum hydroxide used as an adjuvant and adsorbent for the recombinant protein antigen is known to increase efficacy of lipopolysaccharide vaccine in vivo thus interfering endotoxin test. The aim of this study is to determine effect of aluminum hydroxide on the LAL test using the hepatitis B vaccine as a model and to optimize the LAL test condition not to be interfered by aluminum gydroxide. (omitted)

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.132-136
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• 2004

The LAL (Limulus amebocyte lysate) test for the detection and quantification of endotoxin is based on the gelation reaction between endotoxin and LAL from a blood extract of Limulus polyphemus. The test is labor intensive, requiring dedicated personnel, a relatively long reaction time (approximately 1 h), relatively large volumes of samples and reagents and the detection of the end-point is rather subjective. To solve these problems, a miniaturized LOC (lab-on-a-chip) prototype, 62mm (L) ${\times}$ 18 mm (W), was fabricated using PDMS (polydimethylsiloxane) bonded to glass. Using this prototype, in which 2mm (W) ${\times}$ 44.3mm (L) ${\times}$ 100 $\mu\textrm{m}$ (D) microfluidic channel was constructed, turbidometric and chromogenic assay detection methods were compared, and the chromogenic method was found the most suitable for a small volume assay. In this assay, the kinetic-point method was more accurate than the end-point method. The PDMS chip thickness was found to be minimized to around 2 mm to allow sufficient light transmittance, which necessitated the use of a glass slide bonding for chip rigidity. Due to this miniaturization, the test time was reduced from 1 h to less than 10 min, and the sample volume could be reduced from 100 to ca. 4.4 ${\mu}$L. In summation, this study suggested that the LOC using the LAL test principle could be an alternative as a semi-automated and reliable method for the detection of endotoxin.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.242-248
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• 2013

Endotoxins are part of the outer membrane of the cell wall of gram-negative bacteria and are continuously released during bacterial growth. Endotoxins typically induce severe sepsis and septic shock, which cause more than 50% of mortalities. Endotoxins are easily measured in the serum by the limulus amebocyte lysate (LAL) test. However, a nonspecific result is obtained, because the high concentration of serum proteins disturbs the enzyme reaction of the LAL test. In order to solve this problem, the LAL test was performed in this study after the centrifugation of the boiled serum samples to remove the impurities. As a result, among the various conditions examined, endotoxin measurement with the LAL test was the most accurate and repeatable after centrifugation of the boiled serum at $100^{\circ}C$. Moreover, the endotoxin was accurately and repeatedly measured from the prepared sera of mice that had been administered an intraperitoneal injection of purified lipopolysaccharides (LPS) or E. coli. Therefore, the application of centrifugation to remove impurities from boiled serum gives an accurate measurement of endotoxins in the sera of normal subjects or patients, and this will lead to the improved diagnosis and prevention of diseases caused by endotoxins. In addition, the centrifugation of boiled serum samples should be considered and included in the development of endotoxin test kits.

• KSBB Journal
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• v.18 no.5
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• pp.429-433
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• 2003

LAL (Limulus amebocyte lysate) test to detect and quantity endotoxin is based on gellation reaction between endotoxin and LAL from a blood extract of Limulus polyphemus. The test is labor intensive requiring dedicated personnel, takes relatively long reaction time (approximately 1 hr), requires relatively large volume of samples and reagents, and its end-point detection method is rather subjective. To solve these problems, we attempted to develop a miniaturized LOC (lab-on-a-chip) prototype using PDMS and glass. Using the 62 mm (length) ${\times}$ 18 mm (width) prototype in which 2 mm (width) ${\times}$ 44.34 mm (length) ${\times}$ 100 $\mu\textrm{m}$ (depth) microfluidic channel was provided, we compared the various detection methods of gellation, turbidometric, and chromogenic assays to find the chromogenic method to be the most suitable for small volume assay. In this assay, kinetic point method was more accurate than end point method. We also found the PDMS chip thickness should be minimized to around 2 mm to allow sufficient light transmittance, which necessitated a glass slide bonding for chip rigidity. Through the miniaturization, the test time was reduced from 1 hr to less than 10 minutes, and the sample volume could be reduced from 100 ${\mu}\ell$ to 4.4 ${\mu}\ell$. In sum, this study revealed that the mini LOC could be an alternative for a semi-automated and reliable method for LAL test.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.24 no.3
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• pp.310-320
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• 2014

Backgrounds: Endotoxin, which found in the outer membrane of the gram-negative bacteria cell wall, makes up almost all of the lipopolysaccharide(LPS). When people are exposed to endotoxin,it can result in diverse health effects such as an airway irritation and inflammation, fever, malaise, bronchitis, allergic asthma, toxic pneumonitis, hypersensitivity lung disease. Cases among the elderly, children or pregnant can occur more frequently than a healthy adult if they are repeatedly exposed to the existing endotoxin. Therefore, we investigated and assessed the environmental characteristics associated with the airborne endotoxin concentration level in six hospital lobbies. Method: Endotoxin from indoor air in six hospital lobbies was measured by an area sampling method and analyzed according to American Society for Testing and Materials International(ASTM international) E2144-01. Total suspended particulate(TSP), carbon dioxide($CO_2$), temperature and humidity were also measured by using direct reading measurements or airborne sampling equipment at the same time. Environmental characteristics were appropriately divided into two or three groups for a statistics analysis. One-way analysis variable(one-way ANOVA) was used to examine a difference of the endotoxin concentration, depending on the environmental characteristics. In addition, only variables with p-value(p<0.25) were eventually designed to the best model by using multiple regression analysis. Results: The correlation analysis result indicated that TSP(p=0.003) and $CO_2$(p<0.0001) levels were significantly associated with endotoxin concentration levels. In contrast, temperature(p<0.068) and humidity(p<0.365) were not associated with endotoxin concentration. Levels of endotoxin concentration were statistically different among the environmental characteristics of Service time(p=0.01), Establishment of hospital(p<0.001), Scale of hospital(p=0.01), Day average people using hospital(p=0.03), Cleaning time of lobby(p=0.05), Season(p<0.001), and Cleaning of ventilation system(p<0.001) according to ANOVA. Finally, the best model(Adjusted R-square=72%) that we designed through a multiple regression test included environmental characteristics related to Service time, Area of lobby, Season, Cleaning of ventilation system, and Temperature. Conclusions: According to this study, our result showed a normal level of endotoxin concentration in the hospital lobbies and found environmental management methods to reduce the level of endotoxin concentration to a minimum. Consequently, this study recognized to be requirement for the management of ventilation systems and an indoor temperature in order to reduce the level of endotoxin concentration in the hospital lobbies.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.303-307
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• 1991

The hemolyic polypeptide in delta-endotoxin from Bacillus thun'ngiensis subsp. darmstadiensis 73ElO-2 was purified by Sephadex G-IOO gel filtration and DEAE-cellulose ion exchange column chromatography. The purity of hemolysin was confirmed by ouchterlony test and SDS-PAGE. The molecular weight of the purified hemolysin was approximately 64 KDa by SDS-PAGE. The purified hemolysin has not mosquitocidal activity against larvae of Aedes agypti, but hemolytic activity on red blood cells of rat. There is no serological relationship between delta-endotoxin from B. thuringiensis subsp. israelensis and the purified hemolysin from the . strain 73ElO-2.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.393-398
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• 1993

One strain of mosquitocidal Bacillus thuringiensis, H9B, was isolated from soil. The biochemical characteristics and flagella antigenicity of the strain H9B is similar to that of B. thuringiensis subsp. darmstadiensis. The delta-endotoxin of the strain H9B coincided with that of B. thuringiensis subsp. darmstadiensis strain 73E10-2 on agarose double immunodiffusion test. The delta-endotoxin of B. thuringiensis subsp. israelensis contains hemolysin fragment (28 kb) on SDS-PAGE when the delta-endotoxin was solubilized in alkali, while that of the strain H9B does not contain 28 kb protein.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.124-129
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• 2016

Endotoxin, also known as lipopolysaccharide (LPS) produced by the cell wall of gram negative bacteria can be present in any liquid or on any biomaterial. Endotoxin in blood can cause fever and inflammation. In this study, we compared bacterial endotoxin using Escherichia coli O157:H7, Klebsiella oxytoca, Salmonella Typhi, Shigella sonnei and Morganella morganii. Bacteria were cultured for use in the experiment, and diluted to $1.5{\times}10^8CFU/mL$. A check marked sensitivity confirmatory test of the Limulus amebocyte lysate (LAL) reagent was performed to examine the validity. The end point reaction to each bacteria sample was confirmed with 10 fold dilution and then the final reaction end point was confirmed by 2 fold dilution between the dilution step and the upper dilution step. According to the results, in detection of endotoxins in more than 0.015 EU/mL, E. coli O157 was 75~37.5 CFU/mL, K. oxytoca 37.5~18.75 CFU/mL, M. morganii and S. Typhi 3.75~1.875 CFU/mL, and S. sonnei 7.5~3.75 CFU/mL. The resulting value was finally ensured by a confirmation test for the inhibitory factor. Based on this study, conduct of further research on bacterial endotoxin is encouraged.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.71-81
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• 2004

Peri-implantitis could be the result of biomechanical and occlusal overload as well as microbiologic invasion. The dental implant may be more susceptible to dental plaque than the natural tooth, as the predictability of a stable soft tissue attachment complex has not yet been confirmed. With the development of peri-implantitis, the implant surface would be exposed to the oral environment and becomes coated with bacteria. The objective of therapy for this condition is to regain integration of the implant with bone. Since fibroblast adherence to surfaces is impeded by endotoxin, it would seem that decontamination would be desirable to obtain maximum osseointegration. The purpose of this study was to determine whether various chemotherapeutic and mechanical treatments(distilled water, air-powder abrasive, hypersaturated citric acid, tetracycline) can detoxify contaminated titanium implant surface by means of kinetic LAL test. Experimental rough surface titanium disks were fabricated. All of them were divided into two groups(A.a group and P.g group) and each contaminated by A. actinomycetemcomitans and P. gingivalis suspension. Contaminated disks were treated with distilled water, air-powder abrasive, citric acid and tetracycline, and then all disks were placed into LPS-free water for elution. The results were as follows : 1. In A.a group, LPS elute level of all test groups were significantly lower than control group(p<0.05). 2. In A.a group, LPS elute level of test 2, test 3 and test4 groups were significantly lower than that of control group(p<0.05). But, among the test 2, test 3, test4 groups, the significant differences were not detected. 3. In P.g group, LPS elute level of test 2, test 3 and test 4 groups were lower than that of control group(p<0.05). But, among the test groups, the significant differences were not detected. From the result of this study, it would be concluded that air-powder abrasive, hypersaturated citric acid and tetracycline treatments may be effective at reducing endotoxin level on rough titanium implant surfaces, and can be clinically used. But the treatments in peri-implantitis differentially impact osseointegration making one method clinically superior. To gain this knowledges, further molecular biologic and histopathologic studies should be developed.