• Title/Summary/Keyword: Species detection

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Ttrosine Hydroxylase in Japanese Medaka (Oryzias latipes): cDNA Cloning and Molecular Monitoring of TH Gene Expression As a Biomarker (송사리 Tyrosine Hydroxylase: cDNA 클로닝 및 생물지표로서의 TH 유전자 발현의 분자생물학적 추적)

  • Shin, Sung-Woo;Kim, Jung-Sang;Chon, Tae-Soo;Lee, Sung-Kyu;Koh, Sung-Cheol
    • Environmental Analysis Health and Toxicology
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    • v.15 no.4
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    • pp.131-137
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    • 2000
  • The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.

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The Changes of Cerebral Metabolic Parameters, Serum Levels of Neuron-Specific Enolase and S-100$\beta$ Protein During Retrograde Cerebral Perfusion Under Profound Hypothermic Total Circulatory Arrest (초저체온하 완전순환정지 시에 이용되는 역행성 뇌관류의 시간에 따른 뇌대사 지표, 혈청 내 neuron-specific enolase, 및 S-100 베타단백의 변화)

  • 김경환
    • Journal of Chest Surgery
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    • v.34 no.9
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    • pp.653-661
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    • 2001
  • Background: Retrograde cerebral perfusion(RCP) is one of the methods used for brain protection during aortic arch surgery. The author previously published the data, however, for the safety of it, there still remains many controversies. The author performed RCP and checked various parameters to clarify the possibility of early detection of cerebral injury. Material and Method: The author used pigs(Landrace species) weighing 25 to 30kg and performed RCP for 120 minutes. After weaning of cardiopulmonary bypass, we observed pigs for another 120 minutes. Rectal temperature, jugular venous oxygen saturation, central venous pressure were continuously monitored, and the hemodynamic values, histological changes, and serum levels of neuron-specific enolose(NSE) and S100$\beta$ protein were checked. Central venous pressure during RCP was maintained in the range of 20 to 25 mmHg. Result: Flow rates(ml/min) during RCP were 224.3$\pm$87.5(20min), 227.1$\pm$111.0(40min), 221.4$\pm$119.5(60min), 230.0$\pm$136.5(80min), 234.3$\pm$146.1(100min), and 184.3$\pm$50.5(120min). Serum levels of NSE did not increase after retrograde cerebral perfusion. Serum levels of S100$\beta$ protein(ng/ml) were 0.12$\pm$0.07(induction of anesthesia), 0.12$\pm$0.07(soon after CPB), 0.19$\pm$0.12(20min after CPB), 0.25$\pm$0.06(RCP 20min), 0.29$\pm$0.08(RCP 40min), 0.41$\pm$0.05(60min), 0.49$\pm$0.03(RCP 80min), 0.51$\pm$0.10(RCP 100min), 0.46$\pm$0.11(RCP 120min), 0.52$\pm$0.15(CPBoff 60min), 0.62$\pm$0.15(60min after rewarming), 0.76$\pm$0.17(CPBoff 30min), 0.81$\pm$0.20(CPBoff 60min), 0.84$\pm$0.23(CPBoff 90min) and 0.94$\pm$0.33(CPBoff 120min). The levels of S100$\beta$ after RCP were significantly higher than thosebefore RCP(p<0.05). The author could observe the mitochondrial swellings using transmission electron microscopy in neocortex, basal ganglia and hippocampus(CA1 region). Conclusion: The author observed the increase of serum S100$\beta$ after 120 minutes of RCP. The correlation between its level and brain injury is still unclear. The results should be reevaluated with longterm survival model also considering the confounding factors like cardiopulmonary bypass.

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Studies on the Physiological Chemistry of Flower Organ and Seed in Ginseng Plant. IV. Variation of Free Amino Acids in the Flower and Seeds of the $F_1$ Plants of the Combinations Panax ginseng ${\times}$ Panax quinquefolium and Panax ginseng ${\times}$ Panax japonicus. (인삼종자형성에 대한 생리화학적 연구 IV. 고려인삼과 미국인삼 및 고려인삼과 죽절인삼 $F_1$의 화기 및 종자 형성과정에 있어서의 유리아미노산의 소장)

  • Jong-Kyu Hwang;Hee-Chun Yang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.14
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    • pp.165-172
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    • 1973
  • The sterile phenomenon is frequently found in the inter-species hybrids of ginseng as in other plants. It is known that among the hybrids between Panax Ginseng (PG) and Panax Quinquefolium (PQ), and between Panax Ginseng and Paxax Japonicus (PI), PG${\times}$PI is fertile only very rarely, while PG ${\times}$ PQ is always sterile. Therefore, in order to clarify the relationship between this sterility phenomenon and the metabolism of free amino acids, the changes of free amino acids through the formation of the flower organs and seeds of two hybrids, PG ${\times}$ PQ and PG ${\times}$ PI were investigated by thin layer chromatography. The results are summarized as follows: 1. Distinct differences in the quantity and number of free amino acids were recognized between PG ${\times}$ PQ, PG ${\times}$ PI and their parent plants. From the hybrid PG ${\times}$ PQ, 19 kinds of ninhyrin sensitive substances were detected in all. They were (1) 17 amino acids: alanine, valine, leucine, phenylalanine, proline, hydroxy-proline, serine, threonine, tyrosine, aspartic acid, glutamic acid, lysine, arginine, ${\gamma}$-amino butyric acid, ${\beta}$-alanine, cysteic acid and tryptophan, and (2) two amides: asparagine and glutamine. From the hybrid PG ${\times}$ PI, in addition to the above 19 substances, methionine and one unknown substance were detected. 2. Generally, alanine, as partie acid, glutamic acid, cysteic acid and asparagine were detected in large amounts in the two hybrids as in PG, PG and PJ but it was a noticeable fact concerning these two hybrids that the largest quantity of asparagine was found at microspore satge and pollen mature stage. 3. The decrease of cysteic acid in the two hybrids at the red ripened stage was the same as in PQ and PJ but opposite to the change in PG. The detection of methionine in PG ${\times}$ PJ was worthy of notice. 4. The change of proline was conspicuously different from that in their parent plants. It was detected as a trace of color at the micros pore stage while asparagine was detected in the greatest amount at that time. It is well known that the quantity of proline is closely related to the sterility of plant. This fact was also found true in the formation of ginseng seeds. It was reported as well that asparagine accumulated when proline decreased. 5. The deficiency of proline seemed to be closely related with the sterility of hybrids and with the degradation of pollen in anther. 6. The difference in the changes of free amino acids between the selfed lines of PG, PQ and PJ, and their hybrids seemed to be caused by the transformation of gene-action system by hybridization. On these phenomena along with proline metabolim and its physiological role in seed formation further studies are required.

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Development of Prevotella nigrescens ATCC $33563^T$-Specific PCR Primers (Prevotella nigrescens ATCC $33563^T$ 균주-특이 중합효소연쇄반응 프라이머 개발)

  • Song, Soo-Keun;Yoo, So-Young;Kim, Mi-Kwang;Kim, Hwa-Sook;Lim, Sun-A;Kim, Do-Kyung;Park, Jae-Yoon;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.212-220
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    • 2008
  • A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC $33563^T$-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC $33563^T$-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC $33563^T$ and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC $33563^T$. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC $33563^T$. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC $33563^T$, especially with regard to the maintenance of the strain.

Arsenic Speciation and Risk Assessment of Miscellaneous Cereals by HPLC-ICP-MS (HPLC-ICP-MS를 활용한 잡곡의 비소 화학종 및 위해 분석)

  • An, Jae-Min;Hong, Kyong-Suk;Kim, Sung-Youn;Kim, Dae-Jung;Lee, Ho-Jin;Shin, Hee-Chang
    • Korean Journal of Environmental Agriculture
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    • v.36 no.2
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    • pp.119-128
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    • 2017
  • BACKGROUND: Miscellaneous cereal have been largely consumed in Korea as due to their physiological functions beneficial to human health. The cereals are currently a social concern because they have been found to contain heavy metals. Thus, monitoring heavy metals in the cereals is an important requirement for food safety analysis. In this study, we determined arsenic concentration in the cereals randomly harvested from different markets. METHODS AND RESULTS: Inorganic arsenic was determined by ICP-MS coupled with HPLC system. The HPLC-ICP-MS analysis was optimized based on the limit of detection and recover test to reach $0.13-1.24{\mu}g/kg$ and 94.3-102.1%, respectively. The concentrations of inorganic arsenic equivalent to daily exposure were levels of $19.91{\mu}g/day$ in mixed grain, $1.07{\mu}g/day$ in glutinous rice, $0.77{\mu}g/day$ in black brown rice, $0.13{\mu}g/day$ in barley and $0.11{\mu}g/day$ in soybeans. CONCLUSION: The levels of arsenic in miscellaneous cereals were found lower than the recommended The Joint FAO/WHO Expert Committee on Food Additives (JECFA) levels, suggesting that the cereals marketed in Korea are not potential concern in risk assessment.

Natural Occurrence of Aflatoxigenic Aspergillus Species and Aflatoxins in Traditional Korean Fermentation Starters, Meju and Nuruk

  • Woo, So Young;Lee, Sang Yoo;Tian, Fei;Jeong, A-Yeong;Yoo, Cha Nee;Kang, Seung Yoon;Chun, Hyang Sook
    • Journal of Food Hygiene and Safety
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    • v.35 no.5
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    • pp.438-446
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    • 2020
  • Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.

Pathogenicity and PCR detection of Vibrio tapetis in Manila clams, Ruditapes philippinarum (양식 바지락, Ruditapes philippinarum에 대한 Vibrio tapetis의 병원성과 PCR법에 의한 진단)

  • Park, Sung-Woo;Lee, Kyung-Hee
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.39-48
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    • 2005
  • Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

The Study on the Methylmercury Analysis and the Monitoring of Total Mercury and Methylmercury in Fish (어류 중 메틸수은 분석법 확립 및 모니터링)

  • Kim, Hee-Yun;Chung, So-Young;Sho, You-Sub;Oh, Geum-Soon;Park, Seong-Soo;Suh, Jung-Hyuk;Lee, Eun-Ju;Lee, Yoon-Dong;Choi, Woo-Jeong;Eom, Ji-Yoon;Song, Min-Soo;Lee, Jong-Ok;Woo, Gun-Jo
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.882-888
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    • 2005
  • Procedure for analysis of methylmercury in fish was developed, involving addition of HCl, extraction with toluene, and clean-up using L-cystein solution. Obtained extract is analyzed by gas chromatography with electron capture detector using Ulbon HR-Thermon-Hg column. Detection limit and recovery of the method were 0.005mg/kg (expressed as Hg), 98-107 (103%), respectively. Total mercury and methylmercury concentrations in 175 commercial fish samples ranged from [mean-max (mean), unit: mg/kg]: 0.014-1.200 (0.270) and 0.006-0.901 (0.168) in tuna-fish, 0.020-0.934 (0.323) and 0.012-0.553 (0.149) in martin-fish, 0.082-0.782 (0.391) and 0.040-0.436(0.201) in shark, 0,023-0.031 (0.026) and 0,013-0.018 (0.015) in salmon, 0.098-0.193 (0.133) and 0.031-0.015(0.090) in tilefish, and 0,031-0.214 (0.089) and 0.016-0.093 (0.042) in canned tuna respectively. No sample of analyzed fish exceeded 1.0mg/kg wet wt., limit for methylmercury established by Codex. In all species examined, estimated weekly intake was lower than Provisional Tolerable Weekly Intake recommended by the JECFA (the Joint FAO/WHO Expert Committee on Food Additives).

A Study on Heavy Metal Contamination and Risk Assessment of Seaweed and Seaweed Products (해조류와 해조류가공품의 중금속 오염실태 및 위해성평가)

  • Lee, Ji-Yeon;Lee, Myung-Jin;Jeong, Il-Hyung;Cho, Young-Sun;Sung, Jin-Hee;Baek, Eun-Jin;Lee, Eun-Bin;Kim, Hye-Jin;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.447-453
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    • 2019
  • In this study, the contamination of 4 types of heavy metals (lead, cadmium, arsenic, and mercury) was monitored in 80 seaweeds and their processed products, and a food safety assessment was also carried out for these heavy metals. Lead, cadmium and arsenic were analyzed by ICP-OES and mercury was analyzed by mercury analyzer. The detection ranges of heavy metals were found as follows: Pb (N.D-0.802 mg/kg), Cd (N.D-0.759 mg/kg), As (0.134-17.296 mg/kg), and Hg (0.0005-0.0331 mg/kg). Pb and Hg showed no significant differences among seaweeds whereas Cd and As were significantly higher in the species hizikia fusiforme (P<0.05). Food safety assessment from seaweed intake was measured by PTWI (Provisional Tolerable Weekly Intake), PTMI (Provisional Tolerable Monthly Intake), and MADL (Maximum Allowable Daily Body Load) as set by JECFA (Joint FAO/WHO Expert Committee on Food Additives). Pb and Hg were 0.197%, 0.036% of PTWI respectively, while Cd was 1.877% of PTMI and As was 0.619% of MADL. Therefore, it was found that heavy metal levels of seaweed were low and was considered to be safe for consumption.