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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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보육교사의 효능감이 직무스트레스에 미치는 영향에서 정서조절곤란의 조절효과 검증 (The Moderating Effect of Emotional Dysregulation on the Relationship Between Teacher Efficacy and Job Stress of Teachers in Early Childhood Education and Care)

  • 이경숙;채진영;김명식;박진아;이정민
    • 아동학회지
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    • 제37권4호
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    • pp.145-158
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    • 2016
  • Objective: This study investigated the moderating effect of emotional dysregulation on the relationship between teacher efficacy and job stress among teachers in the Early Childhood Education and Care (ECEC). Methods: The participants included 586 ECEC teachers from 99 centers in Seoul, Gyeonggi-do, Daejeon, Chungcheong-do, Jeolla-do, and Gyeongsang-do. The data were analyzed through frequencies, percentages, and Pearson's correlations using SPSS 21.0 (IBM Co., Armonk, NY). To analyze the moderating effect, Ping's (1996) two-step approach was used via AMOS 20.0 (IBM Co., Armonk, NY). Results: The main findings are as follows. First, the mean scores of ECEC teacher efficacy and job stress showed above the average, and the mean score of emotional dysregulation was the nearly average. Second, fit statistics indicated that the proposed model, as revised, provided an acceptable fit to the sample data. This proposed model showed that the emotional dysregulation of teachers in ECEC had a moderating effect on the relationship between teacher efficacy and job stress. Conclusion: These findings imply that the ECEC teachers showed the higher level of self-trust and self-confidence than average regarding their own work, and suffered from the work overload. Also, the positive and supportive working environment would help the ECEC teachers to reduce their emotional dysregulation. In addition, there was a moderating effect of the ECEC teachers' emotional dysregulation on the relationship between teacher efficacy and job stress. These findings imply that the workshop or counselling programs need to be provided to teachers in order to help control their emotion dysregulation and reduce their job stress.

정관운동에 있어서 prostaglandin 의 역할에 관한 연구 (The Study on the Role of Prostaglandin in Contraction of Vas Deferens)

  • 박원규
    • 대한약리학회지
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    • 제19권2호
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    • pp.1-8
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    • 1983
  • Prostaglandin(PG) is ubiquitously distributed in most mammalian tissue and their actions are complicated. Especially in autonomic nervous system, there are evidences indicating that PGs act as neuromodulators i.e., PGs, which are released in the vicinity of autonomic neuroeffector junctions, influence the release and the response of the neurotransmitter. Present study was undertaken to elucidate the interrelationship between $PGF_{2\alpha}$ and adrenergic ${\alpha}_2-receptor$ function in electrical field stimulation induced contractile response of vas deferens in rat. Male rat, weighing 150{\sim}200\;g, was sacrificed and vas deferens was obtained. The isolated vas deferens strip was placed between two platinum electrodes in temperature controlled $(37^{\circ}C)$ muscle chamber containing Tyrode's solution and the electrical field stimulation(EFS) induced contraction was recorded with Grass Polygraph(Model 7) via force displacement transducer (FT .03, Grass). The results are summarized as follows: 1) Electrical field stimulation for 1sec( 1 msec, 40 cps) induced contraction of vas deferens was completely blocked by tetrodotoxin. 2) Bretylium caused marked inhibition of the EFS-induced contraction, hut tyramine and cocaine augmented the contraction. 3) EFS-induced contraction was inhibited or little affected in distal portion of vas deferens by norepinephrine or methoxamine, but the contraction was rather augmented by the ${\alpha}-agonists$ in proximal portion. 4) Clonidine inhibited the EFS-induced contraction proportionally to the concentration in distal portion, which was blocked by yohimbine pretreatment, but in the presence of $PGF_{2\alpha}$ the blockade by yohimbine was reversed. 5) Indomethacin pretreatment reduced the effect of clonidine, but addition of $PGF_{2\alpha}$ after washing-out the indomethacin caused the contraction to the control level. From these results it is suggested that PG synthesis is a necessary step and the PG itself has a permissive role in ${\alpha}_2-adrenoceptor$ action in rat vas deferens.

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Stator Current Processing-Based Technique for Bearing Damage Detection in Induction Motors

  • Hong, Won-Pyo;Yoon, Chung-Sup;Kim, Dong-Hwa
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2005년도 ICCAS
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    • pp.1439-1444
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    • 2005
  • Induction motors are the most commonly used electrical drives because they are rugged, mechanically simple, adaptable to widely different operating conditions, and simple to control. The most common faults in squirrel-cage induction motors are bearing, stator and rotor faults. Surveys conducted by the IEEE and EPRI show that the most common fault in induction motor is bearing failure (${\sim}$40% of failure). Thence, this paper addresses experimental results for diagnosing faults with different rolling element bearing damage via motor current spectral analysis. Rolling element bearings generally consist of two rings, an inner and outer, between which a set of balls or rollers rotate in raceways. We set the experimental test bed to detect the rolling-element bearing misalignment of 3 type induction motors with normal condition bearing system, shaft deflection system by external force and a hole drilled through the outer race of the shaft end bearing of the four pole test motor. This paper takes the initial step of investigating the efficacy of current monitoring for bearing fault detection by incipient bearing failure. The failure modes are reviewed and the characteristics of bearing frequency associated with the physical construction of the bearings are defined. The effects on the stator current spectrum are described and related frequencies are also determined. This is an important result in the formulation of a fault detection scheme that monitors the stator currents. We utilized the FFT, Wavelet analysis and averaging signal pattern by inner product tool to analyze stator current components. The test results clearly illustrate that the stator signature can be used to identify the presence of a bearing fault.

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Conformational Study of Human Serum Albumin in Pre-denaturation Temperatures by Differential Scanning Calorimetry, Circular Dichroism and UV Spectroscopy

  • Rezaei-Tavirani, Mostafa;Moghaddamnia, Seyed Hassan;Ranjbar, Bijan;Amani, Mojtaba;Marashi, Sayed-Amir
    • BMB Reports
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    • 제39권5호
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    • pp.530-536
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    • 2006
  • Thermal conformational changes of human serum albumin (HSA) in phosphate buffer, 10 mM at pH = 7 are investigated using differential scanning calorimetric (DSC), circular dichroism (CD) and UV spectroscopic methods. The results indicate that temperature increment from $25^{\circ}C$ to $55^{\circ}C$ induces reversible conformational changes in the structure of HSA. Conformational change of HSA are shown to be a three-step process. Interestingly, melting temperature of the last domain is equal to the maximum value of fever in pathological conditions, i.e. $42^{\circ}C$. These conformational alterations are accompanied by a mild alteration of secondary structures. Study of HSA-SDS (sodium dodecyl sulphate) interaction at $45^{\circ}C$ and $35^{\circ}C$ reveals that SDS affects the HSA structure at least in three steps: the first two steps result in more stabilization and compactness of HSA structure, while the last one induces the unfolding of HSA. Since HSA has a more affinity for SDS at $45^{\circ}C$ compared to $35^{\circ}C$, It is suggested that the net negative charge of HSA is decreased in fever, which results in the decrease of HSA-associated cations and plasma osmolarity, and consequently, heat removal via the increase in urine volume.

격자기반의 토양수분 추적에 의한 지하수함양량 추정기법 개발 (Groundwater Recharge Assessment via Grid-based Soil Moisture Route Modeling)

  • 김성준;채효석
    • 한국수자원학회논문집
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    • 제33권1호
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    • pp.61-72
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    • 2000
  • 지하수 함양량을 추정하기 위한 방법으로 격자기반의 일변 토양수분 추적기법을 모델링하였다. 본 모델은 GRASS-GIS를 이용하여 모델에 필요한 자료들을 준비하고, 모델은 이들을 ASCII 형태로 받아들여 일별 수행결과(유역출구에서의 유출량, 지표유출 분포도, 토양수분 분포도 등)들을 GRASS상에서 도시하여 주는 형태로 구성하였다. 보청천유역의 일부인 이평교 유역($75.6\;\textrm{km}^2$)을 대상으로 모델의 적용성을 검토하였다. '95년 및 '96년도의 IHP유역 연구보고서 자료를 이용하여 실측 일유출자료와 모의 일유출자료를 비교하였다. 일단위 및 월단위의 유역 토양수분의 변화양상을 공간적으로 확인할 수 있었으며 본 기법을 이용하여 1995년도 보청천의 이평교유역에 대한 지하수 함양량을 계산하였다.

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Methylome Analysis of Two Xanthomonas spp. Using Single-Molecule Real-Time Sequencing

  • Seong, Hoon Je;Park, Hye-Jee;Hong, Eunji;Lee, Sung Chul;Sul, Woo Jun;Han, Sang-Wook
    • The Plant Pathology Journal
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    • 제32권6호
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    • pp.500-507
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    • 2016
  • Single-molecule real-time (SMRT) sequencing allows identification of methylated DNA bases and methylation patterns/motifs at the genome level. Using SMRT sequencing, diverse bacterial methylomes including those of Helicobacter pylori, Lactobacillus spp., and Escherichia coli have been determined, and previously unreported DNA methylation motifs have been identified. However, the methylomes of Xanthomonas species, which belong to the most important plant pathogenic bacterial genus, have not been documented. Here, we report the methylomes of Xanthomonas axonopodis pv. glycines (Xag) strain 8ra and X. campestris pv. vesicatoria (Xcv) strain 85-10. We identified $N^6$-methyladenine (6mA) and $N^4$-methylcytosine (4mC) modification in both genomes. In addition, we assigned putative DNA methylation motifs including previously unreported methylation motifs via REBASE and MotifMaker, and compared methylation patterns in both species. Although Xag and Xcv belong to the same genus, their methylation patterns were dramatically different. The number of 4mC DNA bases in Xag (66,682) was significantly higher (29 fold) than in Xcv (2,321). In contrast, the number of 6mA DNA bases (4,147) in Xag was comparable to the number in Xcv (5,491). Strikingly, there were no common or shared motifs in the 10 most frequently methylated motifs of both strains, indicating they possess unique species- or strain-specific methylation motifs. Among the 20 most frequent motifs from both strains, for 9 motifs at least 1% of the methylated bases were located in putative promoter regions. Methylome analysis by SMRT sequencing technology is the first step toward understanding the biology and functions of DNA methylation in this genus.

비진공법 CuInSe2 태양전지에서 MoSe2의 생성을 억제하기 위한 산화 몰리브데늄 확산장벽 층 (Molybdenum Oxides as Diffusion Barrier Layers against MoSe2 Formation in A Nonvacuum Process for CuInSe2 Solar Cells)

  • 이병석;이도권
    • Current Photovoltaic Research
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    • 제3권3호
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    • pp.85-90
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    • 2015
  • Two-step processes for preparing $Cu(In,Ga)Se_2$ absorber layers consist of precursor layer formation and subsequent annealing in a Se-containing atmosphere. Among the various deposition methods for precursor layer, the nonvacuum (wet) processes have been spotlighted as alternatives to vacuum-based methods due to their potential to realize low-cost, scalable PV devices. However, due to its porous nature, the precursor layer deposited on Mo substrate by nonvacuum methods often suffers from thick $MoSe_2$ formation during selenization under a high Se vapor pressure. On the contrary, selenization under a low Se pressure to avoid $MoSe_2$ formation typically leads to low crystal quality of absorber films. Although TiN has been reported as a diffusion barrier against Se, the additional sputtering to deposit TiN layer may induce the complexity of fabrication process and nullify the advantages of nonvacuum deposition of absorber film. In this work, Mo oxide layers via thermal oxidation of Mo substrate have been explored as an alternative diffusion barrier. The morphology and phase evolution was examined as a function of oxidation temperature. The resulting Mo/Mo oxides double layers were employed as a back contact electrode for $CuInSe_2$ solar cells and were found to effectively suppress the formation of $MoSe_2$ layer.

Chlamydomonas reinhardtii 바이오매스를 이용한 생물학적 수소생산 (Biological hydrogen production using Chlamydomonas reindardtii biomass)

  • 김미선;백진숙;김선창
    • 한국수소및신에너지학회논문집
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    • 제15권4호
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    • pp.309-316
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    • 2004
  • Chlamydomonas reinhardtii UTEX 90 was cultivated with continuous supply of 2% $CO_2$ using TAP media at $25^\circ{C}$ and produced biomass 1.18 g of dry cell weight/L for 4 days. C. reinhardtii algal biomass(CAB) was concentrated to 20 times by volume and converted into hydrogen and organic acids by anaerobic fermentation using Clostridium butyricum. Organic acids in the fermentate of CAB were consecutively used to produce hydrogen by Rhodobacter sphaeroides KD 131 under the light condition. Approximately 52% of starch in the concentrated CAB which had 4-5.8, 24-26 and 6-7 g/L of starch, protein and fat, respectively was degraded by Cl. butyricum at $37^\circ{C}$. During this process, hydrogen and some organic acids, such as formate, acetate, propionate, and butyrate, respectively were produced. Further conversion of the organic acids in anaerobic fermentate of CAB by Rb. sphaeroides KD131 produced hydrogen from the anaerobic fermentate under the illumination of 8 klux using halogen lamp at $30^\circ{C}$. The result showed that hydrogen was evolved by the anaerobic conversion using Cl. butyricum and then by the photosynthetic fermentation using Rb. sphaeroides KD131. It indicated that the two-step conversion process produced the maximum amount of hydrogen from algal biomass which contained carbohydrate, protein, and fat via organic acids.

Development of a Cognitive Level Explanation Model in Brain Injury : Comparisons between Disability and Non-Disability Evaluation Groups

  • Shin, Tae-Hee;Gong, Chang-Bong;Kim, Min-Su;Kim, Jin-Sung;Bai, Dai-Seg;Kim, Oh-Lyong
    • Journal of Korean Neurosurgical Society
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    • 제48권6호
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    • pp.506-517
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    • 2010
  • Objective : We investigated whether Disability Evaluation (DE) situations influence patients' neuropsychological test performances and psychopathological characteristics and which variable play a role to establish an explanation model using statistical analysis. Methods : Patients were 536 (56.6%) brain-injured persons who met inclusion and exclusion criteria, classified into the DE group (DE; n = 300, 56.0%) and the non-DE group (NDE; n = 236, 44.0%) according to the neuropsychological testing's purpose. Next, we classified DE subjects into DE cluster 1 (DEC1; 91, 17.0%), DE cluster 2 (DEC2; 125; 23.3%), and DE cluster 3 (DEC3; 84, 15.7%) via two-step cluster analysis, to specify DE characteristics. All patients completed the K-WAIS, K-MAS, K-BNT, SCL-90-R, and MMPI. Results : In comparisons between DE and NDE, the DE group showed lower intelligence quotients and more severe psychopathologic symptoms, as evaluated by the SCL-90-R and MMPI, than the NDE group did. When comparing the intelligence among the DE groups and NDE group, DEC1 group performed worst on intelligence and memory and had most severe psychopathologic symptoms than the NDE group did. The DEC2 group showed modest performance increase over the DEC1 and DEC3, similar to the NDE group. Paradoxically, the DEC3 group performed better than the NDE group did on all variables. Conclusion : The DE group showed minimal "faking bad" patterns. When we divided the DE group into three groups, the DEC1 group showed typical malingering patterns, the DEC2 group showed passive malingering patterns, and the DEC3 group suggested denial of symptoms and resistance to treatment.