• Journal of Food Hygiene and Safety
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• v.34 no.6
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• pp.557-561
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• 2019

This study investigated the efficacy of DNA extraction methods for real-time PCR detection of foodborne pathogenic bacteria in livestock manure composts. Livestock manure composts were inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Bacillus cereus and incubated in enrichment broth. For DNA extraction, enriched samples were treated following boiling method, by chloroform, C₁₈ powder, and proteinase K. As a result, 4 species of bacteria were detected by real-time PCR when subjected to boiling for 30 min and treated with proteinase K. These results suggest that detection of foodborne pathogens by real-time PCR from livestock manure composts could be applicable using effective DNA extraction methodology such as the boiling method or proteinase K.

• The Korean Journal of Zoology
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• v.29 no.3
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• pp.171-180
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• 1986

The effects of proteases, neuraminidase and EDTA on adhesion of amoebae on the substratum, ultrastructure and biochemical composition of the cell surface were studied by concanavalin A (con A) cytochemistry and SDS PAGE. By con A cytochemistry the glycocalyx of the plasmalemma was easily subdivided into outer filamentous (F) layer and the inner amorphous (A) layer. On treatment with neuraminidase, amoebae attached to the substratum and spreaded better than untreated cells exposing the more con A binding sites in A- and F-layer. When the cells were treated with trypsin or proteinase K, cells stayed unattached for 12 and 48 hr, respectively. Con A binding sites of A layer and all of those glycoproteins were removed by proteinase K. On the other hand, trypsin damaged all of the con A binding sites in both A- and F-layer without significant change in PAS-stained profile of the plasmalemma. Some of the mucopolysaccharides of the cell surface were released by these enzymes and EDTA. When the cells were incubated with monovalent con A they did not attch on the substratum and cytolysed. From these results adhesion of amoebae on the substratum appears to be mediated by the interaction of the glycoproteins and mucopolysaccharides of the A layer.

• Korean Journal of Medicinal Crop Science
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• v.12 no.1
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• pp.47-52
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• 2004

The root of Patrinia villosa Jussieu (Valerianaceae) has long been used for treatment of infectious diseases in Korea. In this study, the anti-inflammatory effect of the Patrinia villosa root water extract (PVWX) was investigated in proteinase-activated receptor-2 (PAR2)-mediated rat paw edema. Paw edema was induced by injection of trypsin or $trans-cinnamoyl-LIGRLO-NH_2\;(tc-NH_2)$ into hindpaw of rats. PVWX. (10, 50, 100 and 200 mg/kg) was orally administered 1 h before the induction of inflammation. At doses of 50, 100 and 200 mg/kg, PVWX. showed significant inhibition on both change in paw volume and vascular permeability. PVWX. (100 mg/kg) significant1y inhibited PAR2 agonists-induced myeloperoxidase (MPO) activity in paw tissue. These results indicate that PVWX has an anti-inflammatory action in PAR2-mediated paw edema.

• Korean Journal of Food Science and Technology
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• v.22 no.5
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• pp.569-574
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• 1990

The influence of NaCl and pH on degradation of chicken breast muscle myofibrillar proteins by porcine leukocyte lysosomal proteinases was investigated. The degradation reactions were carried out at $38^{\circ}C$ for 24hours under different conditions. The degradation of myofibrillar proteins by leukocyte lysosomal enzymes at various pH values was limited to partial hydrolysis. Reactions at higher pH values resulted in lower molecular weight degradation products while reactions at lower pH resulted in higher molecular weight degradation products. When NaCl was added into the reaction mixture, enzyme activities of degradation were increased at all pH values studied, as evidenced by NPN-analysis and SDS-PAGE. More severe degradation was observed with higher salt concentration. The concentration of 0.5M NaCl in the reaction mixture gave more degradation of myosin heavy chain by enzyme than that of 0.1M NaCl.

• BMB Reports
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• v.41 no.11
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• pp.814-819
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• 2008

Interleukin-32 (IL-32) induces a variety of proinflammatory cytokines and chemokines. The IL-32 transcript was reported originally in activated T cells; subsequently, it was demonstrated to be abundantly expressed in epithelial and endothelial cells upon stimulation with inflammatory cytokines. IL-32 is regulated robustly by other major proinflammatory cytokines, thereby suggesting that IL-32 is crucial to inflammation and immune responses. Recently, an IL-32$\alpha$-affinity column was employed in order to isolate an IL-32 binding protein, neutrophil proteinase 3 (PR3). Proteinase 3 processes a variety of inflammatory cytokines, including TNF$\alpha$, IL-$1{\beta}$, IL-8, and IL-32, thereby enhancing their biological activities. In the current study, we designed four PR3-cleaved IL-32 separate domains, identified by potential PR3 cleavage sites in the IL-32$\alpha$ and $\gamma$ polypeptides. The separate domains of the IL-32 isoforms $\alpha$ and $\gamma$ were more active than the intrinsic $\alpha$ and $\gamma$ isoforms. Interestingly, the N-terminal IL-32 isoform $\gamma$ separate domain evidenced the highest levels of biological activity among the IL-32 separate domains.

• Parasites, Hosts and Diseases
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• v.26 no.2
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• pp.95-106
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• 1988

Specific or non-specific cytolytic processes of free-living amoebae causing meningoencephalitls have been emphasized and the cytolytic ability related to hydrolases in Entantoeba sp. and Naegleria sp. has also been reported since the latter half of 1970's. However, no information on hydrolase activities in Acanthamoeba sp. is available. Hydrolases in Acanthamoeba culbertsoni, a pathogenic species of free-living amoebae, were assayed and compared with those in a non-pathogenic species, A. royreba. Pathogenicity of these two species was confirmed through experimental infection to BALB/c mice. Hydrolase activities and cytotoxic effects between pathogenic and non.pathogenic species were compared in the trophozoites cultured in CGV media and in CHO cell line, respectively. The results are summarized as follows: 1. The mice infected with A. culbertseni were all dead 15 days after nasal inoculation, and the mean survival time was 8.5 days. Also the mice infected with this pathogenic species manifested typical meningoencephalitis, whereas the mice infected with A. royreba did not. 2. Hydrolases detected both in the cell extracts and culture media were acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$, neutral proteinase and acid proteinase, all of which were detected with remarkably higher rate in A. culbertsoni than in A. royreba. 3. A. cuzbertsoni revealed strong cytotoxicity for the target CHO cells, whereas A. royreba did not show any specific cytotoxicity. About 80% of the target cells mixed with A. culbertsoni were dead 48 hours after cultivation, and more than 95% of the target cells were dead 72 hours after cultivation. 4. Hydrolase activities in A. culbertsoni cultured with the target cell line were assayed according to the culture time. The activities of acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$ and acid proteinase in this pathogenic amoeba were detected higher in amoeba extracts than in culture media up to 120 hours after cultivation, but after 120 hours of cultivation those activities were detected higher in culture media than in the amoeba Iysates. Neutral proteinase activity in A. culbertsoni increased more in EBSS medium than in the Iysate specimens although the activity in the extracts was generally steady according to the cultivation time. Summarizing the above results, it is concluded that there were differences in hydrolase activities between Pathogenic A. culbertsoni and non-pathogenic A. royreba, and that some hydrolase activities were detected remarkably higher in A. culbertsoni which revealed strong cytotoxicity to the target CHO cell line.

• Journal of Nutrition and Health
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• v.6 no.1
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• pp.39-46
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• 1973

This study was projected to get basic data which can provide a basis for future direction in nutritional education, and also to find the way how to improve the nutritional supply by evaluating the current nutritional intake of average high school students through the survey study of their daily packed lunch. Five hundred twenty seven students from two boys high school and two girls high school including one general and one vocational school respectively were chosen as random sampling technique. Four hundred forty nine among the 527 students had brought lunch. The contents of lunch box were weighed and converted into nutritional values according to the food composition table and compared with recommended dietary allowances. The results compared and classified by sex, School and housewives' educational level were as follows: 1. The nutritional supply in the lunch box was 671 Cal of energy and 22.3 gm of protein for male students which were respectively 55.9% and 74.2% of the dietary recommendations. On the other side female student's lunch boxes were found to contain 495 Cal of energy and 21.3gm of protein which are respectively 61.8% and 80% of the dietary prescriptions. Excluding niacin, all vitamins and minerals were found to be short. 2. Calorie intake in the vocational high school was found to be higher than in the general high school but lower in protein intake especially significant difference (P<0.01) in animal protein. 3. From the nutritional point of view the educational backgrouud of the housewives was not found to have any influence in the way of preparing the lunch boxes. 4. Nutrients of lunch box were heavily inclined to grain rather than to side dishes.

• Tuberculosis and Respiratory Diseases
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• v.40 no.1
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• pp.43-51
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• 1993

Background: The polymerase chain reaction (PCR) is a very sensitive method for the detecting of mycobacterial DNA. There are many reports revealing the efficacy of PCR for the diagnosis of M. tuberculosis, but there are many different methods for DNA extraction from Mycobacterium tuberculosis. Bead beater method is a very useful method for DNA extraction from clinical spectimens, but its procedures are relatively complicated and time-consuming. So we studied other methods for the DNA extraction from Mycobacterium tuberculosis $H_{37}Rv$ and some clinical specimens (5 smear positive sputa and 5 smear negative CSF). Method: We extracted the mycobacterial DNA with 6 different methods from H37Rv strain and clinical specimens. The methods included SDS-microwave oven method, NaOH lysis method, Triton X-100-Proteinase K method, Lysis buffer method, SDS-proteinase K method and bead beater method. The target DNA was 123bp of IS6110 and was detected by examination of ethidium bromide-stained agarose gels. Results: Among 6 methods, SDS-proteinase K method, bead beater method, lysis buffer method and triton X-100-proteinase K method were excellent, but SDS-proteinase K method was the best method in the aspect of simplicity and cost-effectiveness. Conclusion: We suggest that SDS-porteinase K method is a simple and convinient method and might be the best method for the extraction of mycobacterial DNA.

• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.145-150
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• 2004

Cabbage plants were transformed with the potato proteinase inhibitor II (PINII) gene, bar gene, and hpt gene using Agrobacterium. The expression of the PINII gene was driven by its own promoter which was wound-inducible. Ten transgenic plants were obtained from medium containing hygromycin as a selection antibiotic. The integration and expression of PINII and bar genes were confirmed by Southern and Northern hybridization. Growth and development of diamondback moths (Plutella xylostella) and tobacco cutworm (Spodoptera litura) larvae were examined on $T_1$ plants. The weight of the larvae and pupae of these two insects grown on transgenic plants was not different compared to those grown on wild type plants. However, the pupation and emergence rate of diamondback moths and tobacco cutworms fed on some transgenic plants was lower than on wild type plants. These results suggest that the PINII transgene under the control of a wound-induced promoter may be used for control of insects in transgenic cabbage through reduction of insect progeny number.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.14-19
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• 2000

Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.