• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.1
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• pp.43-49
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• 2002

We have cloned cDNAs encoding toxin from the spider, Araneus ventricosus, and constructed a recombinant baculovirus expressing the insecticidal toxin. The cDNAs encoding toxin were cloned from the cDNA library of A. ventricosus. Sequence analysis of the cDNAs encoding the toxin of A. ventricosus revealed that the 240 bp cDNA for AvTox-1 and 192 bp cDNA for AvTox-2 have an open reading frame of 80 and 64 amino acid residues, respectively. The deduced protein sequence of the toxin genes of AvTox-1 and AvTox-2 was aligned to that of the snack Anemonia sulcata and scorpion Centruroides limpidus limpidus, respectively. Northern blot analysis indicated that AvTox-2 toxin gene showed a fat body-spe-cific expression pattern at the transcriptional level. Furthermore, we have explored the possibility of improving baculovirus by incorporating the A. vontricosus toxin gene into Bombyx mori nuclear polyhedrosis virus genome under the control of polyhedrin promoter, The AvTox-2 toxin gene was expressed as approximately 5.8 kDa band in the recombinant baculovirus-injected silkworm larvae. Bioassays with the recombinant virus expressing AvTox-2 on 5th instar silkworm larvae demonstrated a decrease in the time to kill $(LT_{50} days)$ compared to wild-type BmNPV-Kl $(LT_{50} 6.72 days)$ in the injection of 10 viruses. These results indicate that A. ventricosus toxin is a novel member of the spider toxin family, suggesting that the toxin gene can be used in recombinant baculoviruses to reduce insect feeding damage and increase the speed of insect kill.

• Journal of fish pathology
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• v.11 no.1
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• pp.43-50
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• 1998

In order to determine whether the tumor-inducing iridoviruses and the mortality-associated iridoviruses from cultured marine fish in Korea belong to same type, we compared the immunological characteristics of these viruses. The electrophoretical pattern of structural proteins of the tumor-inducing was different from that of the mortality-associated iridoviruses. The antigenicity of structural proteins of these viruses were identified by Western blotting using two monoclonal antibodies against tumor-inducing iridovirus. Two monoclonal antibodies recognized a 150 kDa structural protein of tumor-inducing iridoviruses showed. However, the structural proteins of the mortality-associated iridoviruses did not react with these monoclonal antibodies. These results demonstrate that the antigenicity of the structural proteins of tumor-inducing iridovirus is different from that of mortality-associated iridovirus, indicating that these two iridoviruses belong to different types.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.35-43
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• 2020

Objective: The aim of this research was to evaluate the effect of different concentrate levels in diets based on cactus Opuntia Stricta (Haw.) Haw cladodes on the performance of lactating Girolando cows. Methods: The experiment involved 10 Girolando multiparous dairy cows at 512.6 kg of body weight (BW) and producing 13.2 kg milk/d, allocated into two 5×5 Latin squares. The experimental treatments consisted of control diet composed by cactus Nopalea cochenillifera. Salm-Dyck. cladodes (Nopalea), forage sorghum silage and concentrate at 20% on dry matter (DM) basis, and four concentrate levels diets (20%, 24%, 28%, and 32%) plus cactus Opuntia stricta (Haw.) Haw. cladodes (Opuntia) and forage sorghum silage. Results: Regarding cows fed control diet, the nutrients intake were greater than for cows fed with cactus Opuntia and concentrate. Regarding concentrate levels, intakes of DM, organic matter (OM), crude protein (CP), non-fiber carbohydrates (NFC), and total digestible nutrients of cows increased linearly. Organic matter, CP, and NDF digestibilities were similar in between to control diet and cactus Opuntia-based diets. The digestibility of NFC increased linearly when the concentrate was inserted. The N balance was the same for control diet and cactus Opuntia-based diets, irrespective the concentrate levels. Conclusion: For cows producing 14 kg/d with 3.5% of fat, it is recommended 32% of concentrate to be included in cactus Opuntia-based diets, and the increase in concentrate level promotes a linear increase in milk yield.

• Imaging Science in Dentistry
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• v.38 no.1
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• pp.39-47
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• 2008

Purpose: The aim of this study was to observe a direct effect of irradiation on the periodontopathic Porphyromonas gingivalis (P. gingivalis). Materials and Methods: P. gingivalis 2561 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Changes in viability and antibiotic sensitivity, morphology, transcription, and protein profile of the bacterium after irradiation were examined by pour plating method, disc diffusion method, transmission electron microscopy, RT-PCR, and immunoblot, respectively. Results: Viability of irradiated P. gingivalis drastically reduced as irradiation dose was increased. Irradiated P. gingivalis was found to have become more sensitive to antibiotics as radiation dose was increased. With observation under the transmission electron microscope, the number of morphologically abnormal cells was increased with increasing of irradiation dose. In RT-PCR, decrease in the expression of fimA and sod was observed in irradiated P. gingivalis. In immunoblot, change of profile in irradiated P. gingivalis was found in a number of proteins including 43-kDa fimbrillin. Conclusion: These results suggest that irradiation may affect the cell integrity of P. gingivalis, which is manifested by the change in cell morphology and antibiotic sensitivity, affecting viability of the bacterium.

• Korean Journal of Veterinary Research
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• v.43 no.2
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• pp.247-253
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• 2003

Actinobacillus pleuropneumoniae causes a highly contagious pleuropneumoniae in swine. The bacterium produces several virulence factors such as exotoxin, LPS, capsular polysaccharide, etc. Among them, the exotoxin, called Apx, has been focused as the major virulence factor, and the toxin consists of 4 gene cluster. apx CABD. apxA is the structural gene of toxin and has four different types, I, II, III, and IV. As the first step of development of a new subunit vaccine, the three different types of apxA gene were amplified from A. pleuropneumoniae isolated from Korea by PCR with primer designed based on the N- and C-terminal of the toxin. The sizes of apxIA, IIA and IIIA were 3,073, 2,971 and 3,159bps, respectively. The comparison of whole DNA sequences of apxIA, IIA and IIIA genes with those of the reference strain demonstrated 98%, 99% and 98% homology, respectively. In addition, the phylogenetic analysis was performed based on the amino acid sequences compared with 12 different RTX toxin family using the neighbor-joining method. ApxA proteins of Korean isolates were identical with reference strains in this study. All ApxA proteins were expressed in E. coli with pQE expression vector and identified using Western blot with polyclonal antibodies against culture supernatants of A. pleuropneumoniae serotype 2 or 5. The sizes of each expressed ApxA protein were about 120, 110, 125 kDa (M.W.), respectively. The results obtained in this study could be used for the future study to develop a new vaccine to porcine pleuropneumoniae.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1484-1489
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• 2014

A novel esterase gene, est01, was successfully unearthed from a biogas digester microbiota metagenomic library. The 1,194 bp est01 gene encodes a protein of 44,804 Da (designated Est01). The amino acid sequence of Est01 shows only moderate (33%) identity to a lipase/esterase. Phylogenetic analysis and biochemical characterization confirmed that Est01 is a new member of family VIII esterases. The purified Est01 from recombinant Escherichia coli BL21 (DE3) showed high hydrolytic activity against short-chain fatty acid esters, suggesting that it is a typical carboxylesterase rather than a lipase. Furthermore, the Est01 was even active at $10^{\circ}C$ (43% activity remained), with the optimal temperature at $20^{\circ}C$, and had a broad pH range from 5.0 to 10.0, with the optimal pH of 8.0. These properties suggest that Est01 is a cold-adaptive esterase and could have good potential for low-temperature hydrolysis application.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.33-43
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• 2013

A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

• Journal of the Korean Society of Food Culture
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• v.30 no.1
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• pp.77-85
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• 2015

This study investigated the physicochemical properties and antioxidative activity of soymilk added with buckwheat sprout (SBS), and sensory properties of SBS were analyzed with control soymilk (CS). Moisture content decreased while protein, ash, and lipid contents increased according to content of buckwheat sprout. Solids content and viscosity tended to increase with addition of buckwheat sprout from 0% (CS) to 2.5% (2.5SBS), 3.5% (3.5SBS), and 4.5% (4.5SBS). SBS was found the significant pH drop from 7.08 to 6.43, 6.34, and 6.21. Suspension stability of soymilk slightly decreased with addition of buckwheat sprout. Hunter's color value L of SBS decreased while a and b values increased in comparison with CS. Rutin content was measured 23.78 mg/100 g (2.5SBS), 39.68 mg/100 g (3.5SBS), and 44.80mg/100 g (4.5SBS). Vitamin C content in SBS was higher than CS. Daidzin content increased as buckwheat sprout was added. Total phenolic content increased from 100.95 mg/100 g (CS) up to 315.71 mg/100 g (4.5SBS). For free radical scavenging activity, SBS was significantly higher than CS. In conclusion, SBS can be used as a functional food with higher amounts of rutin, vitamin C, total phenolics, and daidzin. 2.5SBS and 3.5SBS also showed better overall quality characteristics.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.3
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• pp.319-324
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• 2014

Soybean (Glycine max (L.) Merr.) is an important crop for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. But, antinutritional factors in the raw mature soybean are exist. Raffinose and stachyose are main antinutritional factors in soybean seed. Both raffinose and stachyose are carbohydrates, belonging to the raffinose family of oligosaccharides (RFOs). RFOs are not readily digested in humans and cause flatulence or diarrhea. The objective of this research is to obtain the information on raffinose and stachyose content according to genotype and environment. A total of twenty two soybean genotypes (11 cultivars, 3 germplasms and 8 breeding lines) were selected. Each genotype was grown in the field for two years with two replications and harvested in bulk at natural maturity for two years. Content of raffinose and stachyose was detected by HPLC. The raffinose content (g/kg) of 22 genotypes was $2.68{\pm}0.21-5.87{\pm}2.43$ in year 1 and was $3.24{\pm}0.37-9.05{\pm}0.16$ in year 2. The stachyose content (g/kg) was $4.23{\pm}0.98-27.68{\pm}9.90$ at year 1 and was $5.11{\pm}1.09-25.32{\pm}0.35$ in year 2. Genotype and environment have highly significant effects on raffinose and stachyose content. Three genotypes (Da-7, 116-13, and RS-78) have low stachyose content at 5% significant level in two years. A positive correlation ($R^2=0.1985^*$) between raffinose and stachyose was observed in year 2. These informations are valuable in soybean genetics and breeding program related with raffinose and stachyose content.

• Tuberculosis and Respiratory Diseases
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• v.59 no.2
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• pp.142-150
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• 2005

Background : Continuous growth stimulation by various factors, as well as chronic oxidative stress, may co-exist in many solid tumors, such as lung cancer. A new family of antioxidant proteins, the peroxiredoxins (Prxs), have been implicated in the regulation of many cellular processes, including cell proliferation, differentiation and apoptosis. However, a real pathophysiological significance of Prx proteins, especially in lung disease, has not been sufficiently defined. Therefore, this study was conducted to investigate the distribution and expression of various Prx isoforms in lung cancer and other pulmonary conditions. Method : Patients diagnosed with lung cancer, and who underwent surgery at the Ajou Medical Center, were enrolled. The expressions of Prxs, Thioredoxin (Trx) and Thioredoxin reductase (TR) were analyzed using proteomic techniques and the subcellular localization of Prx proteins was studied using immunohistochemistry on normal mouse lung tissue. Result : Immunohistochemical staining has shown the isoforms of Prx I, II, III and V are predominantly expressed in bronchial and alveolar lining epithelia, as well as in the alveolar macrophages of the normal mouse lung. The isoforms of Prx I and III, and thioredoxin were also found to be over-expressed in the lung cancer tissues compared to their paired normal lung controls. There was also an increased amount of the oxidized form of Prx I, as well as a putative truncated form of Prx III, in the lung cancer samples when analyzed using 2-dimensional electrophoresis. In addition, a 43 kDa intermediate molecular weight protein band, and other high molecular weight bands of over 20 kDa, recognized by the anti-Prx I antibody, were present in the tissue extracts of lung cancer patients on 1-Dimensional electrophoresis, which require further investigation. Conclusion : The over-expressions of Prx I and III, and Trx in human lung cancer tissue, as well as their possible chaperoning function, may represent an attempt by tumor cells to adjust to their microenvironment in a manner advantageous to their survival and proliferation, while maintaining their malignant potential.