• Journal of Environmental Science International
• /
• v.6 no.1
• /
• pp.69-73
• /
• 1997

Endotoxin from the cell wall of marine V. vujnificus was .extracted using the hot phenol-water method, injected endotoxin into rat, and tested the toxic effect of endotoxin on the blood component In rat blood. The results showed that blood glucose, blood urea nitrogen, white blood cell, and reticulocyte were Increased and red blood cell was the same as the number of control group(normal blood), but platelet was decreased. Above results suggested that endotoxin induced a malfunction of liver and that the Increase of white blood cell was for the removal of foreign toxic substance.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.7
• /
• pp.727-733
• /
• 2009

Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

• BMB Reports
• /
• v.41 no.11
• /
• pp.820-825
• /
• 2008

Bacillus thuringiensis Cyt2Aa toxin is a mosquito-larvicidal and cytolytic $\delta$-endotoxin, which is synthesized as a protoxin and forms crystalline inclusions within the cell. These inclusions are solubilized under alkaline conditions and are activated by proteases within the larval gut. In order to assess the functions of the N-and C-terminal regions of the protoxin, several N- and C-terminal truncated forms of Cyt2Aa were constructed. It was determined that amino acid removal at the N-terminal, which disrupts the $\beta$1 structure, might critically influence toxin production and inclusion formation. The deletion of 22 amino acids from the C-terminus reduced the production and solubility of the toxin. However, the removal of more than 22 amino acids from the C-terminus or the addition of a bulky group to this region could result in the inability of the protein to adopt the proper folding. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the production and folding of the B. thuringiensis cytolytic $\delta$-endotoxin.

• Journal of environmental and Sanitary engineering
• /
• v.8 no.1
• /
• pp.129-138
• /
• 1993

The marked spread of the cockraches of recent years has raised a great social problem in urban areas. The cockroach have to remove1 because transmit a disease to human as pest insect, but particulars are not yet reported on biological control agent for the cockroach removal. This study was tried for the first time on biological control for the cockroach removal. The obtained results were as follows : 1. The isolated were spore-forming bacillus 1098 strain in soil. The No. 109(TH 109) strain of the among spore-forming bacillus was showed the poisonous against Cockroach. 2. The biological characteristics and flagella antigenicity of the strain is similar to Bacillus thur-ingiensis subsp. indiana. 3. TH 109 strain have the delta-endotoxin of cuboid shap. 4. This delta-endotoxin of product by TH 109 strain was toxic to the cockroach(Blattella gemzanica. L).

• Journal of Microbiology and Biotechnology
• /
• v.6 no.3
• /
• pp.189-193
• /
• 1996

Submicron-sized polymeric particles (SSPP) were used to remove nucleic acids and endotoxins from cell lysates. The positively charged SSPP selectively adsorb nucleic acids and endotoxins and form complexes with them. The complexes can be easily removed by sedimentation or centrifugation. The removal of nucleic acids and endotoxins using SSPP also can be accomplished in the presence of cell and cell debris. Therefore, nucleic acids and endotoxins can be removed in an initial step of the down-stream processes. In bakers yeast and E. coli lysate systems, the level of DNA could be reduced more than three orders of magnitudes and endotoxins more than seven orders of magnitudes concurrently willi the cell debris removal process using SSPP.

• BMB Reports
• /
• v.42 no.7
• /
• pp.462-466
• /
• 2009

Lipopolysaccharide (LPS), found in the outer membrane of Gram negative bacteria, only exerts its toxic effects when in free form. LPS has three major parts, lipid A, the toxic component, along with a core polysaccharide and O-specific polysaccharide. LPS monomers are known to have molecular masses between 10 to 30 kDa. Under physiological conditions, LPS exists in equilibrium between monomer and vesicle forms. LPS removal by 100 kDa ultrafiltration was more efficient (99.6% of LPS removed) with a low concentration of protein (2.0 mg/ml) compared to a high concentration (20.1 mg/ml). In the presence of different detergents (0.5% Tween 20, 1.0% taurodeoxycholate and 1.0% Triton X-100), LPS removal was more efficient at low protein concentrations (2.0 mg/ml) compared to high protein concentrations (20.1 mg/ml).

• Journal of Periodontal and Implant Science
• /
• v.30 no.2
• /
• pp.429-442
• /
• 2000

Based on current evidence in the literature, it is known that endotoxin is a weakly adherent surface phenomenon and that power-driven instruments can be used to accomplish definitive root detoxification and maximal wound healing without overinstrumentation of root and without extensive cementum removal. And one of the newly developed curette tips used with low power of piezoelectric ultrasonic scaler, is effective to remove calculus and not to remove the excessive cementum. The purpose of this study is therefore, to assess the influence of ultrasonic power and various working parameters on root substitute removal when instrumentation is performed with the curette tip on piezoelectric ultrasonic scaler. This study assessed defect depth, width and area resulting from instrumentation using a piezoelectric ultrasonic scaler with a curette type tip in vitro to acrylic resin block as a root substitute. The working parameters was standardized by the sledge device which controls lateral force(0.5 N, 1 N, 2 N) and instrumentation time(5 sec, 10 sec, 20 sec) and power setting was adjusted 0,2,4,8 in P mode. Power setting had the greatest influence on defect depth compared to lateral force and instrumentation time(standardized regression parameter estimates${\pm}$standard error, $0.37{\pm}0.02$, $0.19{\pm}0.02$, $0.07{\pm}0.02$). The effects on defect area also greatest for power setting($0.57{\pm}0.03$) compared to lateral force and instrumentation time($0.33{\pm}0.03$, $0.12{\pm}0.03$). The effect of the power setting on the defect width($0.15{\pm}0.01$) is not so great as defect depth or defect area compared to lateral force($0.12{\pm}0.01$) and effect of instrumentation time is minimal($0.02{\pm}0.01$). It could be concluded that the power setting has the greatest influence on the defect depth and area in curette type tip with low power of piezoelectric ultrasonic device. Many parameters can be adjusted in various situation in clinical use of piezoelectric ultrasonic scaler but the power setting is the first parameter to be adjusted.

• Journal of Periodontal and Implant Science
• /
• v.27 no.1
• /
• pp.205-215
• /
• 1997

In periodontal regeneration treatment, access to the frucation area is very difficult. Thus complete removal of plaque, calculus and endotoxin is somewhat impossible. In this study, teeth that were extracted due to periodontal disease were used. The furcation area was treated with periodontal curette, ultrasonic scaler, roto bur and they observed using SEM. The result was follows 1. The group treatment with curette showed remaining plaque, the cementum existed in most of the surface and partial dentinal tubule orifice could be seen. 2. The group treatment with ultrasonic scaler showed less removalof plaque compared to curette and irregular surface could be seen. 3. The group treatment with roto bur showed cleaner surface and many dentinal tubule orifice could be seen compared to the curette and ultrasonic scaler groups. Thus when suing treatments such as bone grafting or guided tissue regeneration, it is considered that the furcation area should be treatment with Roto bur.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.12
• /
• pp.2156-2164
• /
• 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

• Journal of Periodontal and Implant Science
• /
• v.24 no.1
• /
• pp.131-143
• /
• 1994

For maintenance of exposed implant in healthy state, it is necessary to treat the surface of implant fixture and provide the surface adjustable to surrounding tissues. Variable techniques have been introduced such as citric acid and air-abrasive system to treat the failed implant. Although when the rough surface of HA coated implant was exposed to oral environment, the surface treatment method with citric acid or air-abrasive system is effective for removal of bacterial endotoxin, it is unsuccessful to prevent plaque deposition due to difficulty in removal of rough surface of HA coated implant. Thus, in this study the method that removes bacterial endotoxin and makes smooch surface without alteration of surface characteristics was studied. HA coated disc manufactured by IMZ Co. Was treated with high speed diamond bur, low speed diamond bur, stone bur, rubber point, jetpolisher. And then its surface state was examined with profilometer and SEM to evaluate the surface smoothness, and its surface component was analyzed with EDX to evaluate wheter the surface characteristics were altered or not. As a result, following results were obtained. When the surface roughness of each implant disc was measured by profilometer, the group I showed a $R_{max}\;2.11{\mu}m$ and the group II, III, IV, V showed a $R_{max2}\;4.17{\mu}m$, $7.28{\mu}m$, $8.61{\mu}m$ and $39.44{\mu}m$ respectively. That is, surface smoothness was highest in the group I and it has been gradually decreased in the group II, III, IV and V. Under the SEM examination, the group I showed relatively smooth surface and the group II showed slightly rougher surface than the group I due to partially remaining HA particles while most HA particle was removed. The group III and IV showed rough topography due to HA particles that was not grinded, and HA coated surface in group V showed very irregular surface with deep groove and prominence. In cross-sectional view, the group I showed uniform surface, and the group III, IV showed rough surface due to remaining HA particles but the thickness of HA coating was remarkably reduced. The group II has similar pattern in group I, and the group V showed about $40{\mu}m$ thickness although it was not constant. By analysis of surface component with EDX, the group II in which the grinding was effective showed a small quantity of calcium and phosphorous and the group III, IV, in which the grinding was incomplete showed calcium and phosphorus peak. In all experimental group, no other than titanium, aluminum, calcium, phosphorus was observed.