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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Determination and Survey of Fluoroquinolones Residue in Chicken Muscle by HPLC with Fluorescence Detector (액체크로마토그래피-형광검출기를 이용한 닭고기 중 플루오로퀴놀론계 항균물질 정량분석 및 잔류조사)

  • 박은정;임지흔;이성모
    • Journal of Food Hygiene and Safety
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    • v.19 no.1
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    • pp.12-18
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    • 2004
  • Ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin in chicken muscle were seperated by liquid extraction and determined with high performance liquid chromatography (HPLC) with fluorescence detector. Analysis was carried out using following conditions; Cl8 column (250${\times}$4.6 mm i.d. 5 ${\mu}{\textrm}{m}$ particle size), mobile phase composed of D.W. (containing 0.4% triethylamine and phospholic acid): methanol : acetonitrile (800:100:100, v/v/v), isocratic pump at a flow rate of 1.0 $m\ell$/min and 50 ${mu}ell$ of injection volume, fluorescence detector with EX278 nm/EM.456 nm. The calibration curves of four fluoroquinolones showed linearity (${\gamma}$$^2$$\geq$0.999) at concenration range of 0.025-0.6 $\mu\textrm{g}$/ml. The recoveries in fortified chicken muscle represented more than 80% with low coefficient of variation (〈10%) for concentration range of four fluoroquinolones. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin were 23.5, 3.4, 3.0 and 2.5 ng/g in chicken muscle, respectively. We also monitored fluoroquinolones residue in muscle of chickens (broiler 1:227, Korean native chicken 219, laying chicken 77) using EEC-4-plate screening and HPLC conformation methods. Ten(broiler 5, Korean native chicken 5) out of the fifteen samples which were positively detected by EEC-plate screening method from 1,523 chicken meat were confirmed with ciprofloxacin and enrofloxacin by HPLC. The ranges of residual concentration were 0-0.12 ppm for ciprofloxacin and 0.01-6.79 ppm for enrofloxacin. In conclusion, our method could be applied effectively to determine four fluoroquinolones residues in chicken meat, and further survey for fluoroquinolones residue in chicken meat are needed for more effective control of fluoroquinolones used in livestock.

A Study on Establishing a Standardized Process for the Development and Management of Food Safety Health Indicators in Korea (우리나라 식품안전보건지표의 개발 및 운용과정 정립에 대한 연구)

  • Byun, Garam;Choi, Giehae;Lee, Jong-Tae
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.217-226
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    • 2015
  • This study was conducted to establish a standardized process for developing food safety health indicators. With this aim, we proposed a standardized process, accessed the validity of the suggested process by performing simulations, and provided a method to utilize the indicators. Developing process for domestic environmental health indicators was benchmarked to propose a standardized process for developing food safety health indicators, and DPSEEA framework was applied to the development of indicators. The suggested standardized process consists of an exploitation stage and a management stage. In the exploitation stage, a total of 6 procedures (initial indicators suggestion, candidate indicators selection, data availability assessment, feasibility assessment, pilot study, and final indicator selection) are conducted, and the indicators are routinely calculated and officially announced in the management stage. The exploitation stage is operated by an interaction between a task force team who manages the overall process, and an advisory committee (minimum of 4 in academia, 2 in research, 4 in specialists of Ministry of Food and Drug Safety) who reviews and performs evaluations on the indicators. The standardized process was simulated with 45 initial indicators, and total of 4 indicators (17 detailed indicators) were selected: 'Proportion of domestic fruit/vegetable receiving 'acceptable' in the evaluation of pesticide/herbicide residues', 'Food-borne disease outbreaks', 'Food-borne legal infectious disease incidence', 'Salmonellosis incidence'. Synthetic food safety health index was derived by calculating percent difference with the data from 2010 to 2012. Results showed that when comparing the year 2010 to 2011, and 2011 to 2012, the overall food safety status improved by 10.37% and 9.87%, respectively. In addition, the contribution of indicators to the overall food safety status can be determined by looking into the individual indicators, and the synthetic index may be illustrated to enhance the ease of interpretation to the public and policy makers. In overall, food health safety indicators can be useful in many ways and therefore, attention should be drawn to conduct further studies and establish related legislations.

Residues of ampicillin in blood of cultured olive flounder by oral, injection and dipping administration (Ampicillin의 경구, 주사 및 약욕 투약에 따른 넙치 혈액에서의 잔류량 변화)

  • Jung, Sung Hee;Seo, Jung Soo;Park, Myoung Ae
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.211-219
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    • 2012
  • The residue levels of ampicillin (AM) in cultured olive flounder, Paralichthys olivaceus (average 300g) at $20{\pm}1.0^{\circ}C$ were studied by oral, intramuscular and dipping administration (routes). The concentrations of AM in the plasma were determined by HPLC-UV detector. The average recoveries of AM in spiked samples between 0.01~10 ppm were ranging from 84.45% to 91.26% for plasma. The limit of detection for AM was 0.05 ppm by using this method. Plasma concentrations of AM were determined after oral dosage (10, 20 and 40 mg/kg body weight), intramuscular injection (5, 10 and 20 mg/kg body weight) and dipping (10, 20 and 40 ppm; 1 h). Samples were taken at 1, 3, 5, 10, 15, 24, 30, 48, 96, 144, 216, 264 and 360 h post-administration. In oral dosage of 10, 20 and 40 mg/kg, it's peak concentrations were $3.62{\pm}0.97$, $5.20{\pm}0.70$ and $11.18{\pm}0.87{\mu}g/ml$, respectively at 10 h post-administration, but AM was not measurable at 144, 360 and 360 h post-administration, respectively. In intramuscular injection of 5, 10 and 20 mg/kg, it's peak concentrations were $6.92{\pm}1.29{\mu}g/m1$, $9.89{\pm}2.22{\mu}g/ml$ and $19.85{\pm}2.97{\mu}g/ml$, respectively at 5 h post-administration, but AM was not measurable at 216, 264 and 264 h post-administration, respectively. In dipping of 10, 20 and 40 ppm, it's peak concentrations were $4.39{\pm}1.10$, $9.57{\pm}1.51$ and $11.61{\pm}1.92{\mu}g/ml$, respectively at 3 h post-administration, but AM was not measurable at 264, 264 and 360 h post-administration, respectively. Therefore, the plasma distribution and elimination levels of AM in olive flounder were dosage-dependant manner in all administration routes.

Studies on Microbiological and Chemical Characteristics of Gamma-Irradiated Soybean Powder (감마선 조사된 대두분의 미생물학적 및 이화학적 특성 연구)

  • Kim, Hee-Yun;Song, Kyong-Hee;Hong, Jin-Hwan;Kim, Dong-Sul;Han, Sang-Bae;Lee, Eun-Ju;Kang, Kil-Jin;Chung, Hyung-Wook;Im, Moo-Hyeog;Kim, Chang-Min;Kwon, Yong-Kwan;Chin, Myung-Shik;Song, In-Hwan;Byun, Myung-Woo;Bae, Dong-Ho;Shin, Il-Shik;Ha, Sang-Chul
    • Korean Journal of Food Science and Technology
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    • v.34 no.6
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    • pp.962-968
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    • 2002
  • Effects of gamma-irradiation of 1 to 10 kGy on the microbial growth, contents of amino acids, fatty acids, and free sugars, and changes in acid values in soybean powder were studied. Irradiation doses at $3{\sim}5\;kGy$ inhibited the mold growth completely in two kinds of imported soybean powders. Contents of sulfur-containing amino acids, such as cysteine, in both soybean powders decreased with irradiation, whereas no significant changes in free amino acid and fatty acid contents of both soybean powders were observed. Free sugar contents of stachyose and sucrose in Chinese soybean powder decreased with increasing irradiation dose level, whereas, those of other sugars remained unchanged. Results of this study confirm that $3{\sim}5\;kGy$ irradiation can be safely applied to apply to soybean powder without causing significant quality deteriorations microbiologically and chemically.

Hazard Analysis for the Cultivation Stage of Strawberry Farms for Securing Preliminary Data to Establish the Good Agricultural Practices (농산물우수관리제도 확립의 기초자료 확보를 위한 딸기농장 재배단계의 위해요소 분석)

  • Lee, Chi-Yeop;Lee, Won-Gyeong;Song, Jeong-Eon;Kim, Kyeong-Yeol;Shim, Won-Bo;Yoon, Yo-Han;Kim, Yun-Shik;Chung, Duck-Hwa
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.97-108
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    • 2012
  • Physical, chemical and biological hazards of strawberry farms at the cultivation stage were analyzed to establish the GAP(Good Agricultural Practice) system. Samples were collected from the plants, cultivation environments(water, soil and air), and personal hygiene (hand, glove, and clothes) of three strawberry farms(A, B, and C) and were tested to analyze physical, chemical (heavy metals and pesticide residues), and biological(sanitary indications and foodborne pathogens) hazards. Physical hazards such as insects and pieces of metal and glass were found in the strawberry farms and can be potential bow for strawberry products. Heavy metal and pesticide residue as chemical hazards were detected at levels lower than the regulation limit. In case of biological hazards, total bacteria and coliform were detected at the levels of 1.6~7.3 and 1.3~5.6 log CFU/g, leaf, mL, hand or $100cm^2$. However, Escherichia coli was not detected in all samples. Bacillus cereus and Staphylococuus aureus were detected at levels of ${\leq}$ 1.1~6.1 log CFU and 4.7~5.4 log CFU/g, mL, hand or $100cm^2$, whereas Listeria monocytogenes, E. coli O157 and Salmonella spp. were not detected in all samples. This study demonstrates that various harzards were in strawberry farms at the growing stage. Therefore proper management such as GAP is needed to prevent the occurrence of food poisoning associated with the hazards revealed in this study.

Risk Analysis for the Harvesting Stage of Tomato Farms to Establish the Good Agriculture Practices(GAP) (GAP 모델 확립을 위한 토마토 농장 수확단계의 위해요소 조사 및 분석)

  • Lee, Chae-Won;Lee, Chi-Yeop;Heo, Rok-Won;Kim, Kyeong-Yeol;Shim, Won-Bo;Shim, Sang-In;Chung, Duck-Hwa
    • Journal of agriculture & life science
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    • v.46 no.4
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    • pp.141-153
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    • 2012
  • Samples collected from six tomato farms(A, B, C : soil culture, D, E, F : Nutriculture) located in Gyeongsangnam-do were tested for the analyses of biological(sanitary indications, major foodborne pathogens, fungi), chemical(heavy metals, pesticides) and physical hazards. The highest levels of total bacteria(7.5 log CFU/g) and coliforms(5.0 log CFU/g) in soil culture farms were higher than those of nutriculture farms(total bacteria: 2.5 log CFU/mL, coliforms: 0.6 log CFU/mL). In crops and personal hygiene soil culture farms showed a slightly higher contamination levels. From all farms, the levels of fungi in soil farms were higher than those of nutrient solution. In case of major pathogens, Bacillus cereus and Staphylococcus aureus were detected in all sample with the exception of nutrient solution. Meantime, Escherichia coli, Listeria monocytogenes, E.coli O157 and Salmonella spp. were not detected. For airborne bacteria, soilculture farms showed less contamination than nutriculture farms. A piece of glass and can was confirmed asphysical hazards. Heavy metal(Cd, Pb, Cu, Cr, Hg, Zn, Ni and As) and pesticide residues as chemical hazards were detected, but their levels were lower than the regulation limit. These results demonstrate that potential hazards on harvesting stage of tomato fam were exposed. Therefore, proper management is needed to prevent biological hazards due to cross-contamination, while physical and chemical hazards were in appropriate levels based on GAP criteria.

Reduction of Stem Inside-cavity and Improvement of Flower Quality in Chrysanthemum 'Baekma' by pH Stabilization and Foliar Spray of Ethephon (양액 pH 안정화와 에세폰 살포에 의한 국화 '백마'의 줄기동공 경감과 절화품질 향상)

  • Hwang, In Taek;Cho, Kyung Chul;Kim, Hee Gon;Ki, Gwang Yeon;Yoon, Bong Ki;Choi, Kyung Ju;Lim, Jin Hee;Choi, Sung Ryul;Shin, Hak Ki
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.4
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    • pp.238-243
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    • 2010
  • This study was conducted to investigate effects of nutrient solution pH control agent and foliar spray of ethephon on the reduction of stem inside-cavity and improvement of flower quality in chrysanthemum 'Baekma'. Changes of pH in nutrient solution as affected by the kind of pH control agent showed more settled pattern in the plot of KOH treatment than in others. Plant growth and development such as cut flower length, leaf numbers, cut flower weight and petal number were the greatest in the plot of KOH. However, the size and area of stem cavity was large in KOH rather than $KHCO_3$. As changes in the absorbing patterns of mineral elements as affected by nutrient solution pH control agent in closed system, contents of total nitrate, phosphorus, potassium and calcium by $KHCO_3$ was absorbed into the plant less than KOH, so that it remained a lot of mineral element residues rather than KOH in closed system. Plant growth as affected by the foliar spray of ethephon showed growth retardation effect in the plot of solution diluted to 1 : 500 and growth promotion effect in the plot of solution diluted to 1 : 1,000 or 1 : 2,000. The number of petals was the best in the plot of foliar spray of ethephon solution diluted to 1 : 1,000 before flowering at 45 days showing 331 petals compared to control showing 302 petals. The size and area of stem cavity as affected the foliar spray of ethephon was smaller 1 mm and 7%, respectively, in the plot of solution diluted to 1 : 2,000 before flowering at 30 days than in control. Therefore, treating pH stabilization using KOH after floral initiation stage with the foliar spray of ethephon solution diluted to 1 : 2,000 before flowering at 30 days would help to reduce stem cavity size and improve flower quality in hydroponically grown chrysanthemum 'Baekma'.

Production of Antimicrobial Compounds and Cloning of a dctA Gene Related Uptake of Organic Acids from a Biocontrol Bacterium Pseudomonas Chlororaphis O6 (생물적 방제균 Pseudomonas chlororaphis O6의 길항 물질 생산 및 유기산 흡수에 관련된 dctA 유전자의 클로닝)

  • Han, Song-Hee;Nam, Hyo-Song;Kang, Beom-Ryong;Kim, Kil-Yong;Koo, Bon-Sung;Cho, Baik-Ho;Kim, Young-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.36 no.3
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    • pp.134-144
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    • 2003
  • A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

Nutrient Balance and Vegetable Crop Production as Affected by Different Sources of Organic Fertilizers (유기자원에 따른 양분수지 및 작물생산)

  • Agus, Fahmuddin;Setyorini, Diah;Hartatik, Wiwik;Lee, Sang-Min;Sung, Jwa-Kyung;Shin, Jae-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.1
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    • pp.1-13
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    • 2009
  • Understanding the net nutrient balance in a farming system is crucial in assessing the system's sustainability. We quantified N, P and K balances under vegetable organic farming in a Eutric Haplud and in West Java, Indonesia in five planting seasons from 2005 to 2007. The ten treatments and three replications, arranged in a completely randomized block design, included single or combined sources of organic fertilizers: barnyard manure, compos ts or green manures. The organic matter rates were adjusted every planting season depending on the previous crop responses. The result sshowed that the application of ${\geq}20$ t $ha^{-1}$ barnyard manure per crop resulted in positive balances of N, P, and K, except in the second crops of 2006 where potassium balance were -25 to -11 kg $ha^{-1}$ under the treatments involving cattle barnyard manure, because of low K content of these treatments and high K uptake by Chinese cabbage. Application of 20 to 25 t $ha^{-1}$ of plant residue or 5 t $ha^{-1}$ of Tithonia compost also resulted in a negative K balance. Soil available P increased significantly under ${\geq}25$ t $ha^{-1}$ barnyard manure and that under chicken manure had the highest available P. Accordingly, chicken barnyard manure gave the highest crop yield because of relatively higher N, P, and K contents. Plant residues gave the lowest yield due to the lowest nutrient content among all sources. Reducing the use of barnyard manure to 12.5 t $ha^{-1}$ and substituting it with Tithonia compost, Tithonia green manure or vegetable plant residue compost gave insignificantly different yield compared to the application of 25 t $ha^{-1}$ barnyard manure singly. In the long run, application of 25 t ha-1 cattle, goat, and horse manure or about 20 t $ha^{-1}$ chicken manure is recommendable for sustaining the fertility of this Andisol for vegetable production.