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Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers (역전사 중합효소 연쇄반응(RT-PCR)에 의한 HCV-RNA의 검출 : Biotin 및 방사성옥소 표지 Primer로 구성된 Kit의 이용)

  • Ryu, Jin-Sook;Moon, Dae-Hyuk;Cheon, Jun-Hong;Chung, Yoon-Young;Park, Hung-Dong;Chung, Young-Hwa;Lee, Young-Sang
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.2
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    • pp.220-226
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    • 1994
  • This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR ) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patient's serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was defected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows ; 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79%) of the same group with conventional method. 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possiblity of false positivity and false negativity.

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Comparison Study on Efficacies of Disinfectants and Sanitizers Among Methods for Quantitative Surface Test (살균소독제의 정량적 표면시험방법별 유효성 비교)

  • Kim, Ae-Young;Kim, Yong-Su;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.238-244
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    • 2010
  • Currently, in vitro suspension tests using tubes are used as a authorized test method for sanitizers and disinfectants. However, the methods could not accurately assess the efficacy of sanitizers and disinfectant on the food-contacted surfaces in the field. This study evaluated the effectiveness of 5 kinds of representative sanitizers and disinfectants against E. coli and S. aureus to compare three quantitative surface testing methods that have been internationally standardized. As a result, the ASTM E2111-05 (ASTM(1)) test method obtained 5.18 $\pm$ 0.03 and 5.27 $\pm$ 0.04 log cfu/carrier reduction in dealing with E. coli and S. aureus, respectively, the ASTM E2197-02 (ASTM(2)) test method obtained 4.63 $\pm$ 0.04 and 3.97 $\pm$ 0.03 log cfu/carrier reduction and the CEN EN 13697 test method should 6.14 $\pm$ 0.05 and 5.31 $\pm$ 0.10 log cfu/carrier reduction in clean condition (CEN(1)) but 4.37 $\pm$ 0.02 and 4.06 $\pm$ 0.01 log cfu/carrier reduction in dirty condition (CEN(2)). Among them, CEN(1) showed the highest bactericidal effects, whereas ASTM(2) and CEN(2) revealed low performance (p < 0.05). In conclusion, the bactericidal effects of the ASTM(2) method and the CEN EN 13697 method adopting stainless steel were lower than the ASTM(1) method, which uses glass. The effectiveness assessment results among nationally accredited test methods were different each other. This implies that they could not fit for in the accurate evaluation of sanitization and disinfection on food-contact surfaces in practical food-processing fields. These results could be used as a basic data for establishment of an official surface test methods applicable in the field.

Changes of Browning, Microbiological and Sensory Characteristics of Concentrated Garlic Juices during Storage (마늘 농축액의 저장 중 갈변도, 미생물 및 관능적 특성의 변화)

  • 배수경;김미라
    • Korean journal of food and cookery science
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    • v.14 no.4
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    • pp.394-399
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    • 1998
  • The juice of garlic (Euichun variety) was extracted and concentrated by heating at 90$^{\circ}C$, by using a rotary vacuum evaporator at 45$^{\circ}C$, or by freezing at -50$^{\circ}C$ until the volume was reduced to 70% of the original's. The concentrated garlic juice was packed into 15 ml test tubes wrapped with aluminum foil and kept at 4$^{\circ}C$ or 25$^{\circ}C$ for 60 days. Changes of browning, microbiological and sensory characteristics of the concentrated garlic juices were monitored every 10 days. The specific gravity and viscosity of the prepared juices decreased in the juices concentrated at 90$^{\circ}C$, 45$^{\circ}C$ and -50$^{\circ}C$ in order. Browning of the concentrated garlic juices was slower during the storage at 4$^{\circ}C$ than at 25$^{\circ}C$. Browning occurred rapidly in the juice concentrated at 45$^{\circ}C$ during the storage, especially at 25$^{\circ}C$. The numbers of mesophilic and psychrotrophic bacteria in the juices did not increase significantly during the storage, which means the garlic juices had good shelf-life. The CFUs/ml of garlic juice concentrated at 90$^{\circ}C$ were lower about 1 to 2 log cycles than those in other concentrated juices. The juice concentrated at 90$^{\circ}C$ showed the weakest garlic odor and the strongest cooked odor among the juices. The juice concentrated at -50$^{\circ}C$ had the freshest odor, especially stored at 4$^{\circ}C$, but the juice concentrated at 90$^{\circ}C$ had lowest score in fresh odor. Brown color was dark in the juice concentrated at 45$^{\circ}C$ and green color of all the juices did not change significantly during the storage.

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A Study on the Fouling of Ultrafiltration Membranes Used in the Treatment of an Acidic Solution in a Circular Cross-flow Filtration Bench (순환식 막 모듈 여과장치를 이용한 산성용액의 수처리 공정 시 발생하는 한외여과막 오염에 관한 연구)

  • Kim, Nam-Joon;Choi, Chang-Min;Choi, Yong-Hun;Lee, Jun-Ho;Kim, Hwan-Jin;Park, Byung-Jae;Joo, Young-Kil;Kang, Jin-Seok;Paik, Youn-Kee
    • Membrane Journal
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    • v.19 no.3
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    • pp.252-260
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    • 2009
  • The effects of the treatment of an acidic solution at pH 2 on polyacrylonitrile ultrafiltration (UF) membranes were investigated using a circular cross-flow filtration bench with a membrane module. A substantial reduction in the membrane permeability was observed after 80 hours' treatment of the acidic solution. In addition, the analyses of the sample solutions by ultraviolet/visible absorption spectroscopy and gas chromatography/mass spectrometry (GC/MS), which were taken from the feed tank as a function of the treatment time, showed that a new organic compound was produced in the course of the treatment. From a thorough search of the mass spectral library we presumed the new compound to be 1,6-dioxacyclododecane-7,12-dione (DCD), one of the well-known additives for polyurethane. Based on further experimental results, including the scanning electron microscope (SEM) images and the solid-state NMR spectra of the membranes used for the treatment of the acidic solution, we suggested that the decrease of the permeate flux resulted not from the deformation of the membranes, but from the fouling by DCD eluted from the polyurethane tubes in the filtration bench during the treatment. Those results imply that the reactivity to an acidic solution of the parts comprising the filtration bench is as important as that of the membranes themselves for effective treatments of acidic solutions, for efficient chemical cleaning by strong acids, and also in determining the pH limit of the solutions that can be treated by the membranes.

ANTIMICROBIAL EFFECT OF ANTIBIOTICS AND ROOT CANAL CEMENTS ON THE PREDOMINANT PATHOGENIC ANAEROBIC MICROFLORA IN ROOT CANALS (근관내 주요 혐기성 병인균에 대한 수종 항생제와 근관충전용 세멘트의 항균효과에 관한 연구)

  • Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.18 no.2
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    • pp.515-525
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    • 1993
  • The purpose of this study was to evaluate the susceptibility of anaerobic microorganisms to certain antibiotics and root canal cements. Prevotella intermedia(Bacteroides intermedius) ATCC 25611(serotype A), Fusobacterium nucleatum ATCC 25586, Actinomyces viscosus ATCC 15987 which are the predominant pathogenic anaerobes in dental root canals were cultured in BHI for 48 hours(Fig.1). After each $200{\mu}l$ of those broths with microorganisms was streaked on each surface of blood agar plate, 2 to 5 antibiotic discs which are impregnated with Tetrncycline, Erythromycin, Ampicillin, Clindamycin, or Vancomycin were applied on each surface of blood agar plate and cultured for 5 days anaerobically in the anaerobic chamber (Fig.2). 15 antibiotic discs for each kind of antibiotics and each species of microorganisms were tested. Also each kind of root canal cement tubes which include Zinc oxide eugenol cement, Zinc phosphate cement, Calcium hydroxide powder+DD.W., Calcium hydroxide paste(Pulpdent Tempcanal), or Vitapex(Table 1) were applied on the inoculated BAPs after $200{\mu}l$ of each experimental species of microorganisms was streaked on the surface of blood agar plates, and they were cultured for 5 days anaerobically in the anaerobic chamber(Fig.3). The sensitivity(antimicrobial effect) was determined by the diameter of the inhibition zone. The results are as follows: 1. The results of antibiotic susceptibility test(Table 2) 1) All of the tested antibiotics had antimicrobial activity with various degrees. 2) In Prevotella intermedia (old Bacteroides intermedius), the diameter of inhibition zone to Erythromycin($37.87mm{\pm}2.20$) was largest, those to Tetracycline($26.20mm{\pm}2.96$), Vancomycin($21.53mm{\pm}1.96$), Clindamycin($18.73mm{\pm}0.96$) was smaller than former orderly, and That to Ampicillin ($7.87mm{\pm}0.83$) was smallest. 3) In Actinomyces viscosus, the diameter of inhibition zone to Erythromycin($28.73mm{\pm}1.22$) was largest, those to Ampicillin($21.73mm{\pm}1.03$), Clindamycin($21.33mm{\pm}1.59$) was similarly next order, that to Vancomycin($19.00mm{\pm}1.96$) was smaller than Clindamycin, and that to Tetracycline($11.93mm{\pm}0.70$) was smallest. 4) In Fusobacterium nucleatum, the diameter of inhibition zone to Ampicillin($31.07mm{\pm}1.91$) was largest, that to Erythromycin($28.87mm{\pm}0.92$), Clindamycin($20.47mm{\pm}1.51$), Vancomycin ($16.73mm{\pm}0.96$), Tetracycline ($12.13mm{\pm}1.06$) are smaller than former orderly. 2. The results of root canal cements and pastes(Table 3) 1) The external diameter of tube is 4mm, so 4mm of the inhibition zone diameter means non-susceptable. Prevotella intermedia (old Bacteroides intermedius) was non-susceptable to Calcium hydroxide powder+D.D.W., Calcium hydroxide paste(pulpdent Tempcanal), and Actinomyces viscosus was non-susceptable to Zinc phosphate cement, Calcium hydroxide powder + D.D.W., Calcium hydroxide paste(pulpdent Tempcanal). 2) In Prevotella intermedia (old Bacteroides intermedius), the diameter of inhibition zone to Zinc oxide eugenol cement($13.67mm{\pm}3.30$) was largest, that to Vitapex($9.20mm{\pm}2.96$), Zinc phosphate cement($6.13mm{\pm}2.07$) was smaller than former. 3) In Actinomyces viscosus, the diameter of inhibition zone to Zinc oxide eugenol cement($17.40mm{\pm}5.20$) was largest and that to Vitapex($8.80mm{\pm}1.70$) was next order. 4) In Fusobacterium nucleatum, the diameter of inhibition zone to Vitapex($42.33mm{\pm}17.2$) was largest and those to Calcium hydroxide paste(Pulpdent Tempcanal)($14.47mm{\pm}3.72$) and Zinc oxide eugenol cement($8.93mm{\pm}2.71$), Zinc phosphate cement($8.20mm{\pm}2.27$), Calcium hydroxide powder+D.D.W.($5.53mm{\pm}2.10$)was next orderly. And then In Zinc oxide eugenol cement and Zinc phosphate cement group, two of fifteen samples showed no inhibition zone, in Calcium hydroxide powder + D.D.W. group, 8 of 15 samples showed no inhibition zone.

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Effect of Phosphorous Acid on Control of Phytophthora Blight of Red Pepper (고추 역병에 대한 아인산 (phosphorous acid)의 방제 효과)

  • Lee, Yong-Se;Ryu, Yeon-Ju;Cho, Jeong-Sang;Lim, Tae-Heon;Chang, Tae-Hyun
    • Korean Journal of Environmental Agriculture
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    • v.20 no.3
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    • pp.180-185
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    • 2001
  • Control effects of phosphorous acid were investigated on Phytophthora blight of red pepper plants in greenhouse and field. In vitro test, the mycelial growth of Phytophthora capsici was inhibited by the phosphorous acid more than 97% at 1,000 ${\mu}g$ and 10,000 ${\mu}g$ a.i./mL in the liquid and solid culture, respectively. At concentration of 10 ${\mu}g$ a.i./mL of phosphorous acid, in the liquid culture the mycelial growth of P. capsici was inhibited 46.2%, however inhibited only 4.9% on the soild culture. Zoosporangial formation was also inhibited 89.1% by phosphorous acid at 100 ${\mu}g$ a.i./mL. Phosphorous acid affected more zoosporangial formation of P. capsici than its mycelial growth. At the concentrations of 10,000, 1,000 and 100 ${\mu}g$ a.i./mL of phosphorous acid, germination of zoosporangia was inhibited 100, 84.3 and 44.2%, respectively. Mycelial growth and zoosporangial formation of P. capsici were little affected at the concentration of 10 ${\mu}g$ a.i./mL of phosphorous acid. Cermination of zoosporangia was also little affected at this concentration, however growth of the germ tubes was inhibited and the abnormal mycelial growth was observed. Phosphorous acid suppressed the incidence of Phytophthora blight of red pepper plants up to $77.0{\sim}62.0%$, in greenhouse. Phosphorous acid suppressed the incidence of Phytophthora blight of red pepper plants up to 54.0% at the conventional culture in field. Treatments of phosphorous acid increased up to 113% in height, 135% in number of fruit, and 129% in weight of fruit.

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Mutational Analysis of MECP2 Gene in 34 Rett Syndrome (Rett 증후군 34례의 MECP2 유전자 변이에 관한 연구)

  • Park, Sang Jo;Hwang, Tae Gyu;Son, Byeong Hee;Kim, Chul Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1263-1272
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    • 2002
  • Purpose : Rett syndrome(RTT) is an X-linked dominant neurodevelopmental disorder affecting 1 per 10,000-15,000 female births worldwide. It was initially described by Andreas Rett in 1966. RTT involves developmental regression characterized stereotypic hand movements, tremors, gait apraxia, seizures, deceleration of head growth after the age of 6-18 months. The disease-causing gene was identified as MECP2 on chromosome Xq28. We carried out mutational analysis of MECP2 genes in RTT patients. Methods : Whole blood(5 cc) of 34 sporadic RTT patients was collected in EDTA-anticoagulated tubes. Genomic DNA was extracted from peripheral blood using the E.Z.N.A. blood DNA kit. Four exons of the MECP2 gene were amplified by PCR in 34 Korean with RTT. We carried out PCR divided the exon three into two parts and the exon four into five parts. Primer sequences designed by Amir et al. in 1999 were almost used(AF030876). Sequencing primers used were the same as PCR. DNA sequencing reactions were performed using an ABI 377 DNA sequencer and ABI PRISM dye terminator cycle sequencing reaction kit(Perkin-elmer). The results were compared with the normal DNA sequence(X99686). To confirm the change of sequence on novel mutations, RFLP analysis was performed. Results : The MECP2 mutations were detected in 23(67.6%) of the 34 patients. The mutations consisted of 12 different types including nine missense and three nonsense mutations. Of these, three (L100V, G161E and T311M) mutations were newly identified. Most of the mutations discovered are located within MBD(39.1%) and TRD(39.1%). In this study, three(T158M, R270X, R306C) mutations were identified high frequency. Conclusion : MECP2 gene was also an important cause of Korean RTT patients. MECP2 gene study is an important tool for diagnosis of Korean RTT patients.

Variations of Fish Community and Fish Density on Artificial Reefs (인공어초어장의 어류 군집상과 어획량 변동)

  • LEE Jeong-Woo;KANG Young-Shil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.535-548
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    • 1994
  • Community structures and fish densit were investigated on three different types of artificial reefs, dice, turtle artificial reef and tubes, constructed in the Korean waters. Variations of fish fauna according to type of artificial reef and the proper artificial reef in each area for optimizing harvest were discussed. Fish were captured by trammel gill net during May, June, September and November, 1988 and both identified and counted. Fourty-five fish species were found in the artificial reefs. Of these, Sebastes spp., Hexagrammos otakii, Pleuronectidae, Navodon modestus and Stephanolepis cirrhifer showed high occurrence-frequency. The dominant species groups were coastal settlement, demersal or rock fishes such as Pleuronectidae, Rajiformes, Stephanolepis cirrhifer, Navodon modestus, Hexagrammos otakii and Sebastes spp. in all of the Artificial reefs except the oceanic area of southern waters. Scomber japonicus was predominant in the oceanic area of southern waters. Composition of demersal, rock and pelagic fishes were different depending on the types of artificial reef. Dice artificial reefs were occupied by rock fish, on the other hand turtle artificial reefs were dominated by dermersal fish. Fish density was high at the dice artificial reef in all survey areas except the middle area of Eastern waters, with high fish density evident in the Tube artificial reef. Fish community structures were remarkably different between Dice and Turtle artificial reefs. The Tube artificial reef showed intermediate characteristics between the above two types of artificial reefs. The coastal areas of Southern waters and the middle and southern areas of Western waters revealed similar fish fauna. Results from the oceanic areas of Southern waters were well associated with the middle and southern areas of Eastern waters.

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Morphological Adaptation of Zostera marina L. to Ocean Currents in Korea (한국산 거머리말(Zostera marina L.)의 해류에 대한 형태적 적응)

  • Lim, Dong-Ok;Yun, Jang-Tak;Han, Kyung-Shik
    • Korean Journal of Environment and Ecology
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    • v.23 no.5
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    • pp.431-438
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    • 2009
  • The main purpose of this research is to prepare and provide basic materials for the propagational strategy of eelgrass by investigating on the morphological adaptation of Korean Zostera marina to ocean currents. An eelgrass plant mainly consists of rhizome, leaf sheath, leaves and roots. The rhizome is the horizontal stem of the plant that serves as the backbone from which the leaves and roots emerge. The leaf sheath is the bundle at the base of the leaves that holds the leaves together, protecting the meristem, the primary growth point of the shoot. Leaves originate from a meristem which is protected by a sheath at the actively growing end of the rhizome. As the shoot grows, the rhizome elongates, moving across or within the sediment, forming roots as it progresses. The aggregated leaves from the leaf sheath are found to have two cell layers on one side and multiple layers of airy tissues called aerenchyma on the other. The aerenchyma tissues are developed in multi-layered cell structures surrounding the veins which are formed in the leaf sheath. Generative shoots are made of rhizomes, which are circular or ovoidal, stem, and spathe and spadix. The transverse section of rhizome and the stem and central floral axis is found to be circular, ovoid and in the shape of convex respectively, and the vascular bundle, which is a part of transport system, has one large tube in the center and two small tubes on both sides. The layers of collenchyma cells numbered from 12 to 15 in the stem, and from 7 to 12 in the rhizome. The seed coat is composed of sclereids, small bundles of sclerenchyma tissues, which prevent the influx of sea water from the outside and help endure the environmental stress. In conclusion, alternative multi-layer structure in circular, convex type aggregated leaf base are interpreted to morphological adaption as doing tolerable elastic structure through movement of seawater. The generative shoots develop long slim stem and branches in circular or ovoidal shapes to minimize the adverse impacts of sea current, which can be interpreted as the plant's morphological adaptation to its environment.

The Comparison of Methods Processing Cells Recovered by Bronchoalveloar Lavage (기관지폐포세척액에서 세포수 측정을 위한 처리방법의 비교)

  • Park, Jae-Seuk;Kim, Jae-Yeal;Lee, Gwi-Lae;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.176-183
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    • 1998
  • Background: The total and differential cell count of bronchoalveolar lavage(BAL) fluid are useful assessing activity, prognosis and response to therapy in diffuse interstitial lung disease. But controversy exist as to the appropriate method in processing BAL fluid. Therefore we investigated the effect of gauze filtration, centrifugation and different storage time of BAL fluid on the total and differential cell count. Methods: We obtained BAL fluid from 6 persons with no active lung lesion and divided pooled BAL fluid into several siliconized glass tubes and filtered through 0, 1, 2, 4 folds of cotton guaze(pore size: 1mm), and compared total cell count using hemocytometer after trypan blue staining and differential cell count after Wright-Giemsa staining of cytocentrifuged preparations. And we also counted total and differential cell count after centrifugation(400g for 30 min) and various storage time(2hr, 24hr, and 48hr). Results: There was no difference in total and differential cell count according to folds of gauze filtraion. But without gauze filtration, mucus threads that hampered total and differential cell count were found in 2 cases (33%). Centrifugation resulted in loss of total cell count($24{\pm}18%$) without change in differential cell count. There was no change in total cell count after 2hr storage but significant cell loss was found after 24hr storage time(24hr : $28{\pm}21%$, 48hr : $41{\pm}24%$). However there was no change in differential cell count with 48hr storage time. Conclusion: Total and differential cell count of BAL fluid may be best performed after cotton gauze filtration without centrifugation and within 2 hours.

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