• Title/Summary/Keyword: hybridization(FISH)

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White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

SUPPRESSION OF HYDROGEN CONSUMING BACTERIA IN ANAEROBIC HYDROGEN FERMENTATION

  • Park, Woo-Shin;Jang, Nam-J.;Hyun, Seung-H.;Kim, In-S.
    • Environmental Engineering Research
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    • v.10 no.4
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    • pp.181-190
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    • 2005
  • Severe loss or hydrogen occurred in most anaerobic hydrogen fermentation reactors. Several selected methods were applied to suppress the consumption of hydrogen and increase the potential of production. As the first trial, pH shock was applied. The pH of reactor was dropped nearly to 3.0 by stopping alkalinity supply and on]y feeding glucose (5 g/L-d). As the pH was increase to $4.8{\pm}0.2,$ the degradation pathway was derived to solventogenesis resulting in disappearance of hydrogen in the headspace. In the aspect of bacterial community, methanogens weren't detected after 22 and 35 day, respectively. Even though, however, there was no methanogenic bacterium detected with fluorescence in-situ hybridization (FISH) method, hydrogen loss still occurred in the reactor showing a continuous increase of acetate when the pH was increased to $5.5{\pm}0.2$. This result was suggesting the possibility of the survival of spore fanning acetogenic bacteria enduring the severely acidic pH. As an alternative and additive method, nitrate was added in a batch experiment. It resulted in the increase of maximum hydrogen fraction from 29 (blank) to 61 % $(500\;mg\;NO_3/L)$. However, unfortunately, the loss of hydrogen occurred right after the depletion of nitrate by denitrification. In order to prevent the loss entangled with acetate formation, $CO_2$ scavenging in the headspace was applied to the hydrogen fermentation with heat-treated sludge since it was the primer of acetogenesis. As the $CO_2$ scavenging was applied, the maximum fraction of hydrogen was enhanced from 68 % to 87 %. And the loss of hydrogen could be protected effectively.

Evolutionary Study on the Dark Chub (Zacco temmincki) 1. Geographic Distribution and Seasonal Variation of Two Allelomorphs of MDH (갈겨니(Zacco temmincki)의 진화에 관한 연구 1. MDH 동위효소 2형의 지리적 분포 및 계절적 변이)

  • 양서영;민미숙
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.232-241
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    • 1989
  • Two cytosol malate dehydrogenase allelomorphs are found in the Dark Chub, Zacco temmincki, a freshwater fish inhabiting Korean waters. Mdh-1MM(hereafter referred to as A-type) is distributed along the south and southeastern parts of Korea, whereas the B-type(Mdh-1MS) is found everywhere except the southeast. Several syrnpatric populations can be found in the southern areas with no hybridization between them. The pattern of geographic distribution would indicate that A-type is superior adaptation to colder water temperature; this is collaborated by evidence collected from a sympatric length of the Tongchon stream, Namhae. The relative warmth of the water along this portion of the stream's length was determined its altitude and the time of year. Frequency of B-types found was directly proportional to the relative warmth of the water and inversely proportional to the nurnber of A-types found in the sarne station. Current data tends to suggest that the B-type of Zacco temrnincki first differentiated from Zacco platypus approximately one million years ago (Yang et al., 1984) while A-type subsequently branched from B-type 300,000 years later(Yang and Mm, 1987) through a progressive adaptation toward colder temperature.

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Expression of Cu/Zn Superoxide Dismutase (Cu/Zn-SOD) mRNA in Shark, Schyliorhinus torazame, Liver during Acute Cadmium Exposure

  • Cho, Young-Sun;Ha, En-Mi;Bang, In-Chul;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Journal of Aquaculture
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    • v.18 no.3
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    • pp.173-179
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    • 2005
  • Superoxide dismutase (SOD), an antioxidant enzyme catalyzing the first step for scavenging the reactive oxygen species is important as an early warning indicator to address various biological stresses. For this reason, the monitoring the expressed pattern of SOD gene in fish organs is one of important biomarkers to assess the aquatic pollution caused by many toxic chemicals. Based on the Northern blot hybridization, semi-quantitative and/or realtime RT-PCRs, the alteration of SOD gene transcripts in shark liver was examined during the experimental acute exposures to cadmium. The expression of SOD at mRNA level was up-regulated both by injection (0, 0.5, 1 or 2 mg $CdCl_2/kg$ body weight for 48 hours) and by immersion (0 or $5{\mu}M$ Cd for 0, 1, 4 and 7 days) treatments of cadmium. The transcriptional stimulation of shark SOD gene by cadmium exposure was dependent upon doses and durations: there was a trend toward more increase in higher dose and longer durations of exposure. The hepatic SOD mRNA levels showed also a general agreement with the tissue cadmium concentrations accumulated in immersion exposure. This result may provide useful strategy to develop a fine molecular biomarker at mRNA level for detecting aquatic pollution caused by toxic metals.

Anaerobic Ammonium Oxidation Process in an Upflow Anaerobic Sludge Blanket Reactor with Granular Sludge Selected from an Anaerobic Digestor

  • Tran, Hung-Thuan;Park, Young-Joo;Cho, Mi-Kyeoung;Kim, Dong-Jin;Ahn, Dae-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.199-204
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    • 2006
  • The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with $NH_4^+\;and\;NO_2^-$ as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescence in situ hybridization (FISH) technique. By the last month of operation, the consumed $NO_2^--N/NH_4^+-N$ ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influent-shutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.

The frequency of chromosomal abnormalities and the prenatal cytogenetic analyses for couples with recurrent abortions

  • Choi, Soo-Kyung;Park, So-Yeon;Han, Jung-Yeol;Ryu, Hyun-Mee;Jun, Jong-Young
    • Journal of Genetic Medicine
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    • v.2 no.2
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    • pp.59-63
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    • 1998
  • Between 1988-1998, cytogenetic analyses were performed for 1,476 couples and 162 women with recurrent abortions. We applied GTG-banding, high resolution-banding and FISH (fluorescent in situ hybridization) techniques in this study. The frequency of balanced translocations was 3.6% (112/3114). Of them, 74 cases (2.38%) were reciprocal translocations and 38 (1.22%) were robertsonian translocations. Chromosome aberrations were more frequent in women (80 cases) than in men (32 cases). No phenotypical abnormalities were found in all carriers who had experienced recurrent spontaneous abortions or experienced giving birth to malformed offsprings. Prenatal cytogenetic analyses were carried out on 40 subsequent pregnancies for carrier couples with balanced translocation. The fetal karyotypes showed that 13 cases (32.5%) were normal, 25 (62.5%) were balanced translocations, and two (6%) were unbalanced translocations. It is believed that the frequency of chromosomal abnormalities in patients with recurrent spontaneous abortion is higher than that of the normal population. Most of the fetal samples showed normal karyotypes or balanced translocations matching that of one of their parents. Although the incidence of chromosomal imbalance in the fetuses was relatively low in prenatal cytogenetic analysis, individuals with balanced translocations are predisposed to giving birth to malformed offsprings with partial trisomy or monosomy. Therefore, we recommend the cytogenetic and the prenatal cytogenetic analysis for those who experiences recurrent abortion as well as in case they become pregnant, to prevent the birth of offsprings with chromosomal abnormalities.

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Low Level of TERC Gene Amplification between Chronic Myeloid Leukaemia Patients Resistant and Respond to Imatinib Mesylate Treatment

  • Mohamad Ashari, Zaidatul Shakila;Sulong, Sarina;Hassan, Rosline;Husin, Azlan;Sim, Goh Ai;Wahid, S. Fadilah Abdul
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1863-1869
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    • 2014
  • The amplification of telomerase component (TERC) gene could play an important role in generation and treatment of haematological malignancies. This present study was aimed to investigate copy number amplification status of TERC gene in chronic myeloid leukaemia (CML) patients who were being treated with imatinib mesylate (IM). Genomic DNA was extracted from peripheral blood of CML-IM Resistant (n=63), CML-IM Respond (n=63) and healthy individuals (n=30). TERC gene copy number predicted (CNP) and copy number calculated (CNC) were determined based on $Taqman^{(R)}$ Copy Number Assay. Fluorescence in situ hybridization (FISH) analysis was performed to confirm the normal signal pattern in C4 (calibrator) for TERC gene. Nine of CML patients showed TERC gene amplification (CNP=3), others had 2 CNP. A total of 17 CML patients expressed CNC>2.31 and the rest had 2.31>CNC>1.5. TERC gene CNP value in healthy individuals was 2 and their CNC value showed in range 1.59-2.31. The average CNC TERC gene copy number was 2.07, 1.99 and 1.94 in CML-IM Resistant patients, CML-IM Respond and healthy groups, respectively. No significant difference of TERC gene amplification observed between CML-IM Resistant and CML-IM Respond patients. Low levels of TERC gene amplification might not have a huge impact in haematological disorders especially in terms of resistance towards IM treatment.

The Effects of Human Adipose Tissue-derived Stem Cells on Degenerative Change of Knee in Rabbit Model (가토 모델에서 인체지방유래 줄기세포가 슬관절의 퇴행성 변화에 미치는 영향)

  • Jeong, Ki-Hwan;Kim, Seok-Kwun;Jeong, Jae-Oo;Heo, Jeong;Kwon, Yong-Seok;Lee, Keun-Cheol
    • Archives of Plastic Surgery
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    • v.35 no.6
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    • pp.637-644
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    • 2008
  • Purpose: The survival of bone marrow derived stem cell was reported several times. But the survival of adipose tissue derived stem cells(hASCs) was not mentioned on. We studied the adipose tissue derived stem cell's survival and effect on articular cartilage in rabbits. Methods: Osteoarthritis was induced in twenty New Zealand white rabbits by intraarticular injection of monosodium iodoacetate(MIA). After four weeks, hASCs were also injected into the knee joints space without any vehicle, but the control group received phosphate buffered saline only. The histologic grade of articular cartilage was measured in 4 and 8 weeks after the transplantation of hASC and the viability of injected stem cells measured by Fluorescent in situ Hybridization (FISH) examination. Results: After 4 and 8 weeks from hASCs transplantation, histologic grade was not significantly difference between two groups(p>0.05), and the Y chromosome of the transplanted hASCs was not detected in articular cartilage. Conclusion: We found that direct injection of hASC in joint space didn't work on damaged articular cartilage repair.

한국산 검정망둑(Tridentiger obscurus)과 민물검정망둑(T.brevispinis)의 분류학적 고찰

  • 김종범;양서영
    • Animal Systematics, Evolution and Diversity
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    • v.11 no.3
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    • pp.359-377
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    • 1995
  • Morphological and isozyme analyses were used to examine closely related species of Tridentiger obscurus and \ulcorner: brevispinis, a pair of cryptic species which has intricate problem with regard to species boundary, and to clarify the taxonomic position. In this study, it was revealed from their 17 allopatric streams that there are differences of band ornamentation on the lateral side and three genetic markers between two species. However, genetic relationship between them (S = 0.813, D = 0.192) was within the intraspecific level of similarity. However, at the sympatry of two species no hybrid was found except at Bangjuk stream where the electrophoretic hybrids with slight deficiency of heterozygosity were observed. It is shown that at sympatry there is a finer microenvironmental subdivision between two species at all populations including Bangjuk stream. To manifest the extent of assortative mating, four syrnpatric subpopulations at Bangjuk stream were analyzed for departure from Hardy-Weinberg equillibrium by F-statistics. All subpopulations were deviated significantly from the expectation under random mating. The evidences presented here supports the recognition of Tridentiger obscurus and T. brevispinis as semispecies in the late stage of reproductive isolation

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Piggery Waste Treatment using Partial Nitritation and Anaerobic Ammonium Oxidation (부분질산화와 혐기성 암모늄산화를 이용한 돈사폐수처리)

  • Hwang, In-Su;Min, Kyung-Sok;Lee, Young-Ok
    • Journal of Korean Society on Water Environment
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    • v.22 no.4
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    • pp.599-604
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    • 2006
  • Nitrogen removal with the combined SHARON (Single reactor system for high ammonium removal over nitrite)ANAMMOX (Anaerobic ammonium oxidation) process using the effluent of ADEPT (Anaerobic digestion elutriated phased treatment) slurry reactor with very low C/N ratio for piggery waste treatment was investigated. For the preceding SHARON reactor, ammonium nitrogen loading and removal rate were $0.97kg\;NH_4-N/m^3_{reactor}/day$ and $0.68kg\;NH_4-N/m^3_{reactor}/day$ respectively. In steady state, bicarbonate alkalinity consumption for ammonium nitrogen converted to $NO_2-N$ or $NO_3-N$ was 8.4 gram per gram ammonium nitrogen. The successive ANAMMOX reactor was fed with the effluent from SHARON reactor. The loading and removal rate of the soluble nitrogen defined as the sum total of $NH_4-N$, $NO_2-N$ and $NO_3-N$ in ANAMMOX reactor were $1.36kg\;soluble\;N/m^3_{reactor}/day$ and $0.7kg\;soluble\;N/m^3_{reactor}/day$, respectively. The average $NO_2-N/NH_4-N$ removal ratio by ANAMMOX was 2.41. Fluorescence in situ hybridization (FISH) analysis verified that Candidatus Kuenenia stuttgartiensis were dominate, which means that they played an important role of nitrogen removal in ANAMMOX reactor.