• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.9-14
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• 1989

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

• Parasites, Hosts and Diseases
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• v.26 no.4
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• pp.239-244
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• 1988

In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot

• KSBB Journal
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• v.18 no.3
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• pp.190-196
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• 2003

An analytical system detecting DNA particularly utilizing a concept of membrane strip chromatography initially applied to home-version tests for, such as, pregnancy and ovulation has been developed. We have chosen S. typhimurium as model analyte among food-contaminating microorganisms that occurred in high frequencies, and invA gene, as a detection target, specific to Salmonella species. This gene was able to be amplified by PCR under optimal conditions employing newly designed primers in our laboratory. The PCR product was specifically measured via hybridization between the analyte and a DNA probe, which was a totally different feature from the conventional gel electrophoresis detecting the products based only on the molecular size. It is notable thar the DNA probe sequence was specially designed such that no separation of excess primers present after PCR was required. This was immobilized on a nitrocellulose (NC) membrane via streptavidin-biotin linkage minimizing a steric effect when the hybridization with the amplified DNA took place. The analyrical system detected the microorganism in a concentration of minimum $10^3$ cfu/mL (i.e., 10 cells per system), estimated from the standard curve, 20 to 40 minutes after adding the sample. This sneitivity was approximately 10 times higher than that of gel electrophoresis as an analytical tool conventionally used. Furthermore, the assay was able to be run at room temperature, which would ofter an extra advantage to users.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.3
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• pp.354-361
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• 2002

The aim of this study is to develop and establish rapid, convenient, and accurate method of analyzing salivary IgA against S. mutans semi-quantitatively. Relative salivary IgA titer was calculated as maximum dilution fold of S. mutans protein that was not detected by salivary antibody after measuring relative intensity of the immune blot bands by densitometry. Analyses were performed in caries-experienced and non-experienced children. Mean IgA titer of non-experienced group shows higher level than that of caries-experienced without statistical significance due to high individual variety of antibody titer in non-experienced group: $2^{6.278{\pm}2.260}$ in non-experienced group and $2^{5.730{\pm}0.499}$ in caries-experienced group(p=0.464). Those results suggest that naturally induced salivary IgA antibodies against S. mutans were present in all subjects, but high titer of antibodies were not achieved in caries-experienced group. On the contrary, antibody titer in non-experienced group shows marked individual variations suggesting that antibody production is multifactorial. In conclusion, immune-slot blot method developed in this study would be useful and applicable in semi-quantitative analysis of antibodies.

• Applied Chemistry for Engineering
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• v.30 no.4
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• pp.429-434
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• 2019

A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

• Journal of Food Hygiene and Safety
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• v.36 no.1
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• pp.1-8
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• 2021

The inverted Y-shaped strip detection method based on acetylcholinesterase (AChE) was developed for the rapid detection of organophosphorus and carbamate pesticides. The inactivation of AChE by organophosphorus and carbamate pesticides has been well known. The AChE catalyzes acetylthiocholine into thiocholine having (-) and (+) charges, and the (+) charge results in aggregation of gold nanoparticle (GNP). Malaoxon and carbofuran were used as standard organophosphorus and carbamate for the development of the inverted Y-shaped strip, respectively. In order to optimize the method, various angles of the Y-shaped strip, different types of nitrocellulose membrane, and concentration of AChE were tested as key parameters. The detection limit of the method was 10 ng/mL for both malaoxon and carbofuran pesticides. No cross-reaction was observed to other pesticides such as atrazine, cyanazine, simazine, bifenthrin, boscalid, metalaxyl, and chlorobenzilate. Recoveries from lettuce spiked when known concentrations of malaoxon and carbofuran were found ranging from 96.4 to 100.7% and 81 to 112.7%, respectively. This study suggests that the inverted Y-shaped strip method based on AChE may be a useful tool for the sensitive, specific, rapid detection of organophosphorus and carbamate pesticides in agricultural products.

• Clean Technology
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• v.27 no.2
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• pp.124-131
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• 2021

As declarations of carbon neutrality are spreading throughout the world, much research is being conducted on biodegradable polymers. In this study, nanocellulose, which comprises the largest amount of natural polymer currently available in the world, was proposed as a substitute for non-biodegradable polymers. We chose to modify the surface functional group of crystalline nanocellulose using glycidoxypropyl trimethoxysilane (GPTMS), which is a silane coupling agent, and the product was then used to form a film with low density polyethylene (LDPE). We then conducted measurements using a Fourier transform infrared spectrophotometer (FT-IR) in addition to measuring hydrophilic/lipophilicity of the surface functional group modification of crystalline nitrocellulose as well as that of a polymer composite using the hybrid nanocellulose (H-NC). For compatibility with petroleum-based polymers, the best tensile strength and transparency was found when the H-NC was reacted at pH 14 and 1 wt% compared with LDPE. From the test results, we found that it is possible to modify the surface functional groups of nanocellulose using a silane coupling agent. In addition, the high compatibility of nanocellulose with petroleum-based polymers is expected to help in reaching carbon neutrality by reducing the use of fossil fuels.

• Conservation Science in Museum
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• v.27
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• pp.67-90
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• 2022

A C-46 transport aircraft, which can be thought of as a large cultural heritage item from the modern period, was subjected to paint analysis and conservation treatment in preparation for its exhibition. The C-46 is the first aircraft ever dispatched to overseas combat zones by the Korean Air Force and carried out missions during the Vietnam War. The aircraft is mainly made of aluminum and shows signs of corrosion on its surface, including pitting and etching, as well as gray and white powdery attachments. In the analysis of the paint, diatomite(SiO₂·nH₂O) was confirmed in the red paint, titanium dioxide(TiO₂) was identified in the white paint, black iron oxide(Fe₃O₄) was detected in the black paint, and colcothar(Fe₃O₄) mixed with putty was confirmed in the blue paint. Infrared spectroscopy revealed the use of alkyd resin in the paint on the main body and nitrocellulose in the Taegeuk pattern. During the conservation treatment, mechanical cleaning, such as sanding, was conducted to remove paint and varnish from the surface. Corrosion was removed by sanding and cleaning with chemical solvents, and new paints and varnishes were applied. Through the paint analysis and conservation treatment, the aircraft was made available for exhibition in a stable condition.

• Journal of Dental Rehabilitation and Applied Science
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• v.19 no.2
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• pp.87-96
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• 2003

The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

• Tuberculosis and Respiratory Diseases
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• v.46 no.4
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• pp.473-480
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• 1999

Background: Early diagnosis of tuberculosis is critical, especially in Korea, an area where tuberculosis is endemic. Because antibody responses to some membrane proteins of Mycobacterium tuberculosis are not comparable, and the policy of BCG vaccination and the prevalence of tuberculosis are different from country to country, the usefulness of the serological diagnostic tests is questionable in Korea, even though they have been confirmed to be useful in other countries. In the specific context of Korea, we tried to evaluate the validity of the ICT Tuberculosis Test (ICT), a membrane-based antibody kit that purports to detect the 5 M. tuberculosis complex-specific antigens including 38-kDa protein. Method: 68 patients with tuberculosis were tested : 37 had no history of previous tuberculosis, and 31 were reactivated cases. The control group comprised 77 subjects : 25 healthy adults, 35 hospital workers with frequent contact with tuberculosis patients, and 17 in-patients with non-tuberculous respiratory diseases. Results: The diagnostic sensitivities of the ICT were 87% and 73% in patients with versus without previous history of tuberculosis, respectively. The sensitivities of smear-positive and smear-negative patient groups were 81% and 73%, respectively. Both of the two patients with extrapulmonary tuberculosis tested positive through the ICT. The specificities of the ICT were 88%, 94%, and 94% in healthy adults, hospital workers, and non-tuberculous patients, respectively, with an overall specificity of 92%. Conclusion: It is suggested that when combined with traditional techniques, the ICT is an useful tool for the diagnosis of pulmonary tuberculosis. The procedure is simple, easy to perform, rapid, and needs no equipment. It shows 73% sensitivity and 92% specificity for the diagnosis of tuberculosis.