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Genotype Frequency of Human Papillomavirus Determined by PCR and DNA Sequencing in Korean Women

  • Lee, Kyung-Ok;Seong, Hye-Soon;Chung, Soo-Jin;Jung, Na-Young;Lee, Hye-Jung;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.2
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    • pp.99-104
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    • 2006
  • Human Papilloma viruses (HPVs) are etiological agents for cervical cancer and are classified into low- and high-risk categories. The aim of this study was to determine the frequency of the HPV genotype in the HPV screening test of Korean women using PCR-direct sequencing. Consensus primers of L1 legion were used for the amplification of HPV DNA and the PCR products (450 bps) obtained were analyzed by automatic sequencing. Sequences were compared with those in GenBank by using the BLAST program. Cervical swab samples of 3,978 women (20-73 years) were tested and the average age was 37.6 years. In this study, 1,174 samples were HPV positive out of 3,978 cervical swab samples screened (29.5%) and 136 samples (11.6%) showed a double infection. A total of 1,310 HPV genotypes were analyzed. The HPV positive rate was the lowest in the 20 years group (69.5%) and most of the samples of the > 60 years group were found HPV positive. Among thirty seven different HPV types identified by sequencing, 21 were HPV high risk types and 16 HPV low risk types were 69.8% (914/1,310) and 26.0% (340/1,310), respectively. In HPV high-risk types, 16 (13.21%), was the most frequently found. HPV 53 (9.62%) and 58 (9.24%) were also frequently found. This group was followed by HPV types 70 (5.50%), 33 (4.73%), 66 (4.20%), 18 (4.05%), 52 (4.05%), 31 (3.97%) and 56 (3.51%) in descending order of frequency. Among HPV low-risk types, 62 (4.20%), 6 (3.59%), 81 (3.59%), 84 (3.51%), and 11 (2.6%) were frequently found. In conclusion, PCR-direct sequencing could be used for quick and reliable typing of known and novel HPVs from clinical specimens. This data could be useful for epidemiological study of HPV and it also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.

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Dietary Risk Assessment for Polycyclic Aromatic Hydrocarbons in Foods (식품중 Polycyclic Aromatic Hydrocarbons의 위해성평가)

  • 이효민;윤은경;박경아;김윤희;정소영;권기성;김명철;송인상;이철호
    • Journal of Food Hygiene and Safety
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    • v.19 no.1
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    • pp.1-8
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    • 2004
  • This study was executed to determine the cumulative dietary risk of PAHs exposed by food ingestion. Food samples including barbecued beef, barbecued pork, grilled chicken, ham, bacon and vegetable oil which were collected from food markets. These samples were saponified, extracted and cleaned up to purify PAHs, and then the purified sample solutions were analyzed by HPLC-FL. Generally, the levels of total PAHs in barbecued beef (0.2 ppb), bacon (0.3 ppb), barbecued pork (0.7 ppb), ham (0.8 ppb), and vegetable oil (1.2 ppb) were low, whereas the level of total PAHs in grilled chicken (9.3 ppb) was significantly high. For the exposure assessment of PAHs due to food ingestion, PAHs levels converted into TEQ$_{BaP}$, the average body weight for 20-73 age group and consumed levels of food proposed from report on the National Health and Nutrition Survey were used. The estimated lifetime average daily intake of dietary PAHs was 4.32${\times}$10$^{-4}$ $\mu\textrm{g}$-TEQ$_{BaP}$kg/day as the mean value. The dietary risk adjusted to cancer potency of benzo(a)pyrene as 7.3 (mg/kg/day)$^{-1}$ was 3.44${\times}$10$^{-6}$ based on current data.ata.

The Effect of LDL on Vibrio vulnificus Septicemia (비브리오 패혈증에 미치는 LDL의 영향)

  • Kim, Jong-Hyeon;Kim, Jong-Suk;Yoo, Wan-Hee;Hur, Hyeon
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.213-217
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    • 2006
  • The halophilic bacterium Vibrio vulnificus is known to be a foodborne pathogen that causes septicemia in human. V. vulnificus infection is characterized by the high fatality rates and the primary attack against a person who have underlying diseases such as liver cirrhosis. However, there is no effective treatment for V. vulnificus septicemia except for classical treatments such as antibiotics. Recently, it has been known that lipoprotein (LDL) plays a major role in the protection against infection and inflammation. Consequently in this paper we analyzed the effects of LDL on V. vulnificus septicemia. We purified V. vulnificus cytolysin, a major virulent factor of V. vulnificus infection and measured inhibitory effects of mouse serum, cholesterol, and LDL on its hemolytic activity. Next experiments were performed to investigate whether LDL has a protective role against septicemia induced by V. vulnificus in mice. Intraperitoneal injection of LDL (1mg as protein) into mice 3hr before V. vulnificus $(1\times10^6\;CFU)$ injection, and V. vulnificus -induced lethality was determined. For the determination the relationship between LDL or cholesterol and prognosis, we determined serum levels of cholesterol and lipoprotein from V. vulnificus septicemia patients (n=15) who had visited the Chonbuk National University Hospital in Chonju. V. vulnificus cytolysin -induced hemolysis of mice erythrocytes was completely inhibited by serum, cholesterol, and low-density lipoprotein. V. vulnificus- induced lethality of mice injected with LDL showed only 40% compared to 100% of control. In survival groups (n=4) of V. vulnificus septicemia patients (n=15), their serum LDL and cholesterol revealed normal levels ($153.3{\pm}40.7,\;LDL;\;190.8{\pm}16.3$, Total cholesterol). However, in death groups (n=11) showed very low levels ($35.6{\pm}13.9,\;LDL;\;59.2{\pm}15.1$, Total cholesterol). Our study indicates that cholesterol and LDL are a prognosis indicator of V. vulnificus septicemia as well as an inhibitor of virulent action of V. vulnificus cytolysin. We suggested that the serum levels of cholesterol or LDL would be major index in the treatment and prevention of V. vulnificus septicemia.

Antihypertensive Effect of Peptide from Sardine Muscle Hydrolyzate on Mild Hypertensive Subjects (정어리 단백질 유래 펩타이드의 경중 고혈압 환자에 대한 강압효과)

  • Kawasaki, T.;Seki, E.;Osajima, K.;Yoshida, M.;Asada, K.;Matsui, T.;Osajima, Y.
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 2004.11a
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    • pp.50-58
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    • 2004
  • Valy-Tyrosine (VY) derived from alkaline pretense hydrolyzate of sardine muscles showed the in vitro Angiotensin I converting enzyme (ACE) inhibitory activity and the in vivo antihypertensive effect in SHR. We investigated the antihypertensive effect of the VY on mild hypertensive subjects including subjects with high-normal blood pressure using a randomized double-blind placebo-controlled study. (1) Nineteen subjects (Age 48.9${\pm}$4.3, M/F;18/1) took a 100ml drink either containing 125${\mu}$g of VY or placebo twice daily for 4 weeks. The reductions of the systolic (SBP) and diastolic blood pressure (DBP) were observed in mild hypertensive subjects (n=5) with averages of 17.8${\pm}$2.5 mmHg (p<0.01 vs placebo) and 11.0${\pm}$2.0 mmHg(p<0.05 vs placebo), respectively. Neither SBP nor DBP changed in the subjects of both the placebo group and the high-normal blood pressure group. (2) A randomized double-blind cross-over placebo-controlled study was carried out in 10 mild essential hypertensive subjects (Age 50.6${\pm}$4.6, M/F;10/0). They took a 100ml drink either containing 62.5${\mu}$g of VY or placebo twice daily for 4 weeks alternatively with a 6-week interval. The percent changes in SBP and DBP were -6.9 % and -5.8 % (p<0.05) one week after the VY drink administration, respectively. No adverse effects such as coughing or allergic phenomena could be observed in any of the subjects of drinking VY during the experimental period. These results suggest that the drink containing at least 125${\mu}$g/day of VY may have a significant antihypertensive effect on mild hypertensive subjects without any adverse effects.

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Bactericidal Efficacy of a Fumigation Disinfectant with Ortho-phenylphenol as an Active Ingredient Against Pseudomonas Aeruginosa and Enterococcus Hirae (Ortho-phenylphenol을 주성분을 하는 훈증소독제의 Pseudomonas aeruginosa와 Enterococcus hirae에 대한 살균효과)

  • Cha, Chun-Nam;Park, Eun-Kee;Kim, Yongpal;Yu, Eun-Ah;Yoo, Chang-Yeol;Hong, Il-Hwa;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.60-66
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    • 2014
  • This test was performed to evaluate the bactericidal efficacy of a fumigation disinfectant containing 20% ortho-phenylphenol against Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus hirae (E. hirae). In preliminary tests, P. aeruginosa and E. hirae working culture suspension number (N value) were $2.8{\times}10^8$ and $4.0{\times}10^8CFU/mL$, respectively. And all the colony numbers on the carriers exposed to the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of bacterial test suspentions by pour plate method), 0.5N2 (the number of bacterial test suspentions by filter membrane method) and 0.5N1, respectively. In addition, the mean number of P. aeruginosa and E. hirae recovered on the control-carriers (T value) was $2.8{\times}10^8$ and $3.4{\times}10^6CFU/mL$, respectively. In the bactericidal effect of the fumigant, the reduction number of $2.8{\times}10^8$ (d value) was 6.46 and 5.19 logCFU/mL, respectively. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. With the results from this study, the fumigation disinfectant containing 20% ortho-phenylphenol has an effective bactericidal activity, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with the pathogenic bacteria.

Comparative Evaluation of the VITEK 2 System and Species-specific PCR Methods for the Detection of Vibrio Species Isolated from Shrimp (새우에서 분리된 Vibrio species 동정을 위한 VITEK 2 system방법과 species-specific PCR방법 비교 평가)

  • Lee, Jeong-Min;Lee, Won-Jun;Kim, Min-Ju;Cho, Yong-Sun;Lee, Jin-Sung;Lee, Hyun-Jin;Yoon, Sang-Woo;Kim, Keun-Sung
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.281-288
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    • 2015
  • Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

A Study on the Content of Minerals in Fortified Food (영양강화식품 중 무기질 함량 조사연구)

  • Kim, Myeong-Gil;Kim, Young-Sug;Kim, Young-Su;Lee, Seong-Bong;Ryu, Kyong-Shin;Yoon, Mi-Hye;Lee, Jong-Bok
    • Journal of Food Hygiene and Safety
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    • v.29 no.2
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    • pp.99-104
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    • 2014
  • This study was done to analyze the contents of minerals and to investigate the ratio of measured values to labeled values and to analyze the ratio of calcium to other minerals in 68 specimen with minerals - fortified commercial beverages, noodles, cereals and grain products. Content of calcium, iron and zinc in samples after microwave digestion was analyzed with an ICP-OES. The measured values of calcium were ranged 82.2~293.1% of the labeled values in 38 samples composed calcium - fortified commercial beverages, noodles, cereals and grain products. The measured values of iron and zinc were ranged 83.3~301.0%, 90.1~314.1% of the labeled values in minerals - fortified commercial beverages, noodles, cereals and grain products, 42, 24 samples. The Ca : Fe ratios were 90.55 (50.55~220.64) in fruit & vegetable juice, 850.41 in fruit & vegetable beverage, 553.49 in blended beverage, 179.07 (118.37~238.01) in soy milk, 204.39(41.64~397.52) in noodle, 296.97(121.64~868.88) in fried noodle, 30.89(15.69~62.05) in cereal and 7.73(0.22~49.92) in grain product. The Ca : P ratios were 1.44(0.96~1.98) in fruit & vegetable juice, 1.92 in fruit & vegetable beverage, 1.66 in blended beverage, 4.23(2.25~7.72) in soy milk, 1.14(0.28~1.97) in noodle, 1.88(1.17~2.42) in fried noodle, 1.29(0.87~2.92) in cereal and 0.30(0.06~1.57) in grain product. The Ca : Mg ratios were 1.85(0.87~5.04) in fruit & vegetable juice, 28.72 in fruit & vegetable beverage, 2.97 in blended beverage, 5.27(2.93~9.36) in soy milk, 3.97(1.34~7.57) in noodle, 6.77(4.63~10.78) in fried noodle, 4.40(2.30~12.55) in cereal and 1.17(0.23~7.48) in grain product. These results suggest calcium contents and the ratio of calcium contents to other minerals in calcium-fortified food products should be strictly controlled. Moreover, to avoid problems with Excessive nutrition, there must be initiatives for better understanding on food labelling and nutrition for fortified food.

A Study on Migration of Heavy Metals from Kitchen Utensils Including Glassware, Ceramics, Enamel, Earthenware and Plastics (유리제 등 조리기구 중 중금속 이행에 관한 연구)

  • Choi, Jae-Chon;Park, Se-Jong;Goh, Hyeah;Lee, Ju Yeun;Eom, Mi Ok;Kim, Meehye
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.334-339
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    • 2014
  • The purpose of our study was to investigate the migration level of lead (Pb), cadmium (Cd), antimony (Sb), arsenic (As), hexavalent chromium ($Cr^{6+}$) and mercury (Hg) from cookwares into food simulants and to evaluate the safety of each heavy metals. The test articles for heavy metals were glassware, ceramics, enamel, earthenware, polypropylene and polyethylene cookwares for Pb and Cd, enamel for Sb, earthenware for As, polyethylene and polypropylene cookwares for $Cr^{6+}$ and Hg. All the article samples of 391 intended for contact with foods were purchased in domestic markets. Pb, Cd, Sb and As were analyzed by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), $Cr^{6+}$ by UV visible spectrophotometer and Hg by mercury analyzer. The migration levels of heavy metals in all the samples were within the migration limits of Ministry of Food and Drug Safety (MFDS). As a result of safety evaluation, our results showed that the estimated daily intakes (EDI, mg/kg bw/day) were $9.12{\times}10^{-6}$ and $8.83{\times}10^{-7}$ for Pb and Cd from ceramics and $1.19{\times}10^{-5}$, $1.23{\times}10^{-5}$ and $7.52{\times}10^{-6}$ for Pb, Cd and Sb from enamel. Tolerable daily intakes (TDI, mg/kg bw/day) were established respectively as 0.0036, 0.00081, 0.0021, and 0.0006 for Pb, Cd, As and Hg by JECFA (Joint FAO/WHO Expert Committee on Food Additives), as 0.0060 for Sb by WHO (World Health Organization). When comparing with TDIs, the EDIs accounted for 0.25% and 0.11% for Pb and Cd from ceramics and 0.33%, 1.52% and 0.13% for Pb, Cd and Sb from enamel.

A Study on Dose-Response Models for Foodborne Disease Pathogens (주요 식중독 원인 미생물들에 대한 용량-반응 모델 연구)

  • Park, Myoung Su;Cho, June Ill;Lee, Soon Ho;Bahk, Gyung Jin
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.299-304
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    • 2014
  • The dose-response models are important for the quantitative microbiological risk assessment (QMRA) because they would enable prediction of infection risk to humans from foodborne pathogens. In this study, we performed a comprehensive literature review and meta-analysis to better quantify this association. The meta-analysis applied a final selection of 193 published papers for total 43 species foodborne disease pathogens (bacteria 26, virus 9, and parasite 8 species) which were identified and classified based on the dose-response models related to QMRA studies from PubMed, ScienceDirect database and internet websites during 1980-2012. The main search keywords used the combination "food", "foodborne disease pathogen", "dose-response model", and "quantitative microbiological risk assessment". The appropriate dose-response models for Campylobacter jejuni, pathogenic E. coli O157:H7 (EHEC / EPEC / ETEC), Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio parahaemolyticus, Vibrio cholera, Rota virus, and Cryptosporidium pavum were beta-poisson (${\alpha}=0.15$, ${\beta}=7.59$, fi = 0.72), beta-poisson (${\alpha}=0.49$, ${\beta}=1.81{\times}10^5$, fi = 0.67) / beta-poisson (${\alpha}=0.22$, ${\beta}=8.70{\times}10^3$, fi = 0.40) / beta-poisson (${\alpha}=0.18$, ${\beta}=8.60{\times}10^7$, fi = 0.60), exponential (r=$1.18{\times}10^{-10}$, fi = 0.14), beta-poisson (${\alpha}=0.11$, ${\beta}=6,097$, fi = 0.09), beta-poisson (${\alpha}=0.21$, ${\beta}=1,120$, fi = 0.15), exponential ($r=7.64{\times}10^{-8}$, fi = 1.00), betapoisson (${\alpha}=0.17$, ${\beta}=1.18{\times}10^5$, fi = 1.00), beta-poisson (${\alpha}=0.25$, ${\beta}=16.2$, fi = 0.57), exponential ($r=1.73{\times}10{-2}$, fi = 1.00), and exponential ($r=1.73{\times}10^{-2}$, fi = 0.17), respectively. Therefore, these results provide the preliminary data necessary for the development of foodborne pathogens QMRA.

Investigation of Microbial Contamination in Oenanthe javanica at Postharvest Environments (미나리(Oenanthe javanica) 수확 후 처리 환경에서의 위생지표세균 및 병원성 미생물 오염도 조사)

  • Kim, Yeon Rok;Lee, Kyoung Ah;Choi, In-Wook;Lee, Young-Ha;Kim, Se-Ri;Kim, Won-Il;Ryu, Song Hee;Lee, Hyo Sub;Ryu, Jae-Gee;Kim, Hwang-Yong
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.268-277
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    • 2014
  • This study assessed microbiological hazards at postharvest stage of dropwort farms (A, B, C, D, E, F, G, H, I) located in 4 different areas in Korea. The samples were assessed for sanitary indication bacteria (total aerobic bacteria, coliform, and Escherichia coli) and pathogenic bacteria (Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus). Total aerobic bacteria and coliform in 9 dropwort farms were detected at the levels of 0~7.00 and 0~4.25 log CFU/g, mL, of $100cm^2$. In particular, microbial contamination in worker's hand showed higher than cultivation environment factors. Escherichia coli was detected in several farms of soil, irrigation water, washing water and worker's hand and also, dropwort in these farms was contaminated with E. coli (positive reaction). In case of pathogenic bacteria, B. cereus was detected at the highest levels in soil. S. aureus was detected qualitatively from only one sample of dropwort washed by water. E. coli O157:H7 and L. monocytogenes were not detected. Although dropwort pass through 2 process (trimming and washing), the microbial contamination was not differ significantly before and after which indicates that current washing system was not effect on reduction of microorganism. From these results, the postharvest environment and workers have been considered as cross-contamination factors. Thus, processing equipments and personal hygiene should be managed to reduce the microbial contamination of dropwort. Accordingly management system such as good agricultural practices (GAP) criteria is needed for the safety of dropwort