• Journal of Korean Society of Forest Science
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• v.86 no.4
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• pp.407-414
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• 1997

The resistance of a non-transgenic poplar clone, 'Ogy' and three transgenic poplar lines to the cottonwood leaf beetle, Chrysomela scripta F., was evaluated by in vitro feeding. The lines were transformed with neomycin phosphotransferase II(NPT II) as a selectable marker, proteinase inhibitor II(pin2) as a resistance gene, and CaMV 35S as a promoter. An efficient method of sterilizing the beetle eggs and introducing them into plant tissue cultures was developed. The resistance of the transgenic lines was investigated in terms of effects tin leaf area consumed, insect weight, insect developmental stages, and plantlet root dry weight after feeding. Also, leaf area consumed was examined by leaf age as measured through leaf plastochron index(LPI). The leaf area consumed and insect weight were highly significant between transformants and control, and insect development in vitro was significant among the transgenic lines. Larval infestation was the most severe around LPI 4 to 5 which were young leaves. The system provided a quick, highly controlled method to screen developing transgenic plantlets directly.

• Journal of Sericultural and Entomological Science
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• v.15 no.1
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• pp.9-14
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• 1973

This study has been carried out to investigate proteinase activity of digestive juice and sucrase activity of midgust tissue in the 5th day of the 5th instar influenced by the dietary composition (quantitative ratio between carbohydrate and protein) and tile rearing temperature during the 4th moulting period. The larvae grew on three kinds of semi-synthetic diet. The A-diet has more carbohydrate than the others, the B-diet has carbohydrate in 1 : 2 with protein, and the C-diet has more protein than the others. All the diets were kept at 16$^{\circ}C$, 25$^{\circ}C$ and 33$^{\circ}C$ during the 4th moulting period. Proteinase activity of digestive juice at the 5th day of the 5th instar was analyzed by Anson's hemoglobin method. Sucrase activity of midgut tissue at the 5th day of 5th instar was analyzed by Somogyi-Nelson's method. The results were as follows. 1. The dietary composition influencing contents of blood sugar was not related to the rearing temperature during the 4th moulting period. The contents of blood sugar appeared to increase in A-diet, B-diet and C-diet order, while proteinase and sucrase activity were stronger in C-diet, B-diet and A-diet order. 2. All kinds of diets showed almost the same fact that proteinase activity at 16$^{\circ}C$ was stronger than that at 32$^{\circ}C$. 3. It was found that sucrase activity became gradualy stronger at 32$^{\circ}C$, 25$^{\circ}C$ and 16$^{\circ}C$ in order in all kinds of diets. 4. There was an interaction in proteinase activity between the dietary composition and the rearing temperature in male larval digestive juice during the 4th mouiting period. On the other hand, there was an inter-acion in sucrase activity between the dietary composition and the rearing temperature in both female and male larval midgut tissue during the 4th moulting period.

• Parasites, Hosts and Diseases
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• v.35 no.1
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• pp.39-46
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• 1997

The present study was undertaken to determine whether live T. uaginnlis degrades human secretory IgA, serum If and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite Iysate, and excretory-secretory product (ESP) of T ucginnlis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA, serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the Iysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as I-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. uaginnlis to HeLa cells were decreased when live T. vusinalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T ucginclis may play a part role in host pathogenesis by T. uosinnlis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.45-50
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• 1998

Hypocotyl explants of flowering cabbage were precultured on MS medium without kanamycin and then cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring insect resistantce proteinase inhibitor II(PI-II) gene in MS liquid medium adjusted pH 5.5 for 72hr. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. After 4 weeks of subculture, kanamycin-resistant shoots were obtained from selection medium. Leaves of putative transformants survived on MS selection medium containing 30 mg/L kanamycin. Incoporation of the PI-II gene into flowering cabbage was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled probe for PI-II gene was hybridized to the expected amplified genomic DNA fragment of about 500 by from transgenic flowering cabbage.

• The Korean Journal of Mycology
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• v.14 no.2
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• pp.101-107
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• 1986

The properties of carboxyl proteinase which was contained in Poria cocos (Schw.) Wolf were investigated by means of the purification with 0.65 ammonium sulfate saturation, DEAE cellulose and Sephadex G-75 gel filtration. This enzyme was found to hydrolyze only peptide bond between glutamyl-L-tyrosine of carbobenzoxy-L-glutamyl-L-tyrosine among the synthetic substrates of carbobenzoxy-L-glutamyl-L-tyrosine, hippuryl- L-phenylalanine and hippuryl-L-arginine. This enzyme was inhibited by $Zn^{+2},\;Fe^{+2},\;Ca^{+2},\;CN^{-1},\;P_2O_7^{-4}$ ions, but stimulated by $Hg^{+2}$ ion. Also, this enzyme was inhibited by organic compounds such as L-lysine, L-phenylalanine, hippuryl-L-phenylalanine, diazoacetyl-DL-norleucine methyl ester (DAN) and 1,2-epoxy-3-(P-nitrophenoxy)propane(EPNP). In particular, the activity was inhibited by L-lysine till 20 minutes of preincubation time rapidly, and by DAN in the presence of $Cu^{+2}$ ion more rapidly after 30 minutes than DAN in the absence of $Cu^{+2}$ ion. L-Lysine was found to be a competitive inhibitor and its $K_i$ value was determined to be 0.12 mmole by Dixon plot.

• Biomedical Science Letters
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• v.8 no.3
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• pp.161-165
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• 2002

Apoptosis can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of rat tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. I describe a new modified method for formalin-fixed, paraffin-embedded tissue sections, pretense pretreatment to permeate the tissue sections that involves an TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) is acknowledged as a method of choice in the rapid identification and quantification of the apoptotic cell fraction in paraffin tissue preparations. TUNEL was performed without apoptosis and with apopotosis samples to each of the three concentrations of proteinase K (10, 25, 40 mg/ml) pretreatments. In this study, I show that chemical pretreatments of the tissue sections in proteinase K (25 mg/ml for 15 min at room temperature) considerably enhances the sensitivity of this nick end labelling technique.

• Journal of Chest Surgery
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• v.27 no.5
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• pp.364-373
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• 1994

Alpha 1-Proteinase inhibitor[PI] was known as a major protective enzyme against to excessive hydrolytic and proteolytic reaction. So, it was suggested that Alpha 1-PI may implicated in growth of bronchogenic cancer. This study was undertaken to investigate the role of Alpha 1-PI in local invasion of bronchogenic cancer. Three groups of patients were studied; Preliminary research group of 15 bronchogenic cancer patients, Main research group of 13 bronchogenic cancer patients and Normal control group of 10 nephrectomy donor. Serum Alpha 1-PI level was observed in each group of patients during pre-and postoperative days. Pre-operative serum Alpha 1-PI level in preliminary research group [329.2$\pm$14.21mg/dl]and main research group[406.2$\pm$39.30mg/dl] were higher than in normal control group[236.2$\pm$19.55mg/dl] significantly[p<0.005]. Serial Alpha 1-PI level in each group during pre-and postoperative days shows peaked at 3rd. postoperative day in preliminary and main research group, thereafter decreased gradually. Immunohistochemical study for Alpha 1-antitrypsin[A1AT] was carried out by ABC[avidin-biotin peroxidase complex] method using Alpha-1 antitrypsin DAKOR to tumor tissues of 13 lung cancer patients in main research group. 6 cases[46.2%, squamous cell ca.;5, adenocarcinoma;1] of above 13 cases show positive immunoreactivity for A1AT. In conclusion, alpha 1-PI and elastase are disclosed that have defined actions for lung cancer growing or spreading.

• Applied Biological Chemistry
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• v.33 no.4
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• pp.287-292
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• 1990

Eight(I through VII) of Bowman-Birk proteinase inhibitor have been isolated from soybean with DEAE-Sephadex A-50. Inhibitor VII was a typical BBTI, showing high cysteine content(17%/mole) ud low trypsin to chymotrypsin inhibiting activity(TIA/CIA= 1.0) with the independent reactive site to each enzyme. Dissociation constant of trypsin-BBTI and chymotrypsin-BBTl complexes were $9.17{\times}10^{-9}M$ and $5.14{\times}10^{-8}M$, respectively. Inhibitor Vll was extremely heat stable. Six hours heat treatment at $100^{\circ}C$ caused only 50% decrease in it's original inhibiting activity. Except inhibitor III,6 other isoinhibitors differed from a typical BBTI in TIA/CIA, values, ranging from 3 to 29.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.135-139
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• 1987

Aspergillus sp. MK-24 was cultured at 3$0^{\circ}C$ in the medium consisting of 2% glucose, 0.2% NaNO$_3$, 0.02% $K_2$HPO$_4$, 0.02% MgSO$_4$ 7$H_2O$, 0.02% KCl, and at initial pH of 5.0. The production of the inhibitor on venom proteinase reached to the maximum in 7 days. Sodium nitrate or potassium nitrate as a nitrogen source was favorable. The production of inhibitor was not affected by the addition of most of the inorganic salt used but depressed by lead, zinc, cobalt, mercuric or silver salts.

• Development and Reproduction
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• v.10 no.4
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• pp.263-269
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• 2006

To evaluate the potentiality of industrial use of cysteine proteinase inhibitor (cystatin) of tilapia egg and serum stability of the tilapia cystatin on low temperature storage and heat treatment was studied. When the eggs were stored at $4^{\circ}C$ for 3 days the cystatin activity was not changed much, while the supernatant of egg homogenate lost its cystatin activity significantly, remaining only about 65% of initial activity. When the eggs and serum were subjected to repeated freeze at $-20^{\circ}C$ and thaw at room temperature once a day, the egg cystatin was decreased after 5 cycles of freeze and thaw. However the serum cystatin was not changed by the 5 times repetition of freeze and thaw. More than 80% of egg cystatin activity was remained when the egg was heated at $35^{\circ}C$ for 30 min, but less than 10% was remained when heated at $50^{\circ}C$. On the other hand, the serum cystatin was very resistant to heat, remaining about 74% after heating at as high as $80^{\circ}C$ for 30 min. In summary, the egg cystatin was more stable when stored as intact form of egg rather than as supernatant of homogenate when stored at refrigeration. Egg cystatin was relatively stable against repeated freeze-thaw, and serum was found to be more stable in cysteine proteinase inhibitory activity than egg. Egg cystatin was not very resistant to heat treatment, while serum cystatin was quite resistant to high temperature heat treatment. These results suggest that tilapia egg and serum, especially the serum, would be a useful source for cysteine proteinase inhibitor in surimi production.