• Title/Summary/Keyword: Fluorescence In situ Hybridization

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Analysis techniques for fermented foods microbiome (발효식품의 마이크로바이옴 분석 기술)

  • Cha, In-Tae;Seo, Myung-ji
    • Food Science and Industry
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    • v.50 no.1
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    • pp.2-10
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    • 2017
  • Human have eaten various traditional fermented foods for a numbers of million years for health benefit as well as survival. The beneficial effects of fermented foods have been resulted from complex microbial communications within the fermented foods. Therefore, the holistic approaches for individual identification and complete microbial profiling involved in their communications have been of interest to food microbiology fields. Microbiome is the ecological community of microorganisms that literally share our environments including foods as well as human body. However, due to the limitation of culture-dependent methods such as simple isolations of just culturable microorganisms, the culture-independent methods have been consistently developed, resulting in new light on the diverse non-culturable and hitherto unknown microorganisms, and even microbial communities in the fermented foods. For the culture-independent approaches, the food microbiome has been deciphered by employing various molecular analysis tools such as fluorescence in situ hybridization, quantitative PCR, and denaturing gradient gel-electrophoresis. More recently, next-generation-sequencing (NGS) platform-based microbiome analysis has been of interest, because NGS is a powerful analytical tool capable of resolving the microbiome in respect to community structures, dynamics, and activities. In this overview, the development status of analysis tools for the fermented food microbiome is covered and research trend for NGS-based food microbiome analysis is also discussed.

Analysis of the Activated Sludge of a Municipal WWTP by Several Bio-Parameters

  • Cho Sun-Ja;Jung Yong-Ju;Park Tae-Joo;Lee Sang-Joon
    • Journal of Environmental Science International
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    • v.14 no.9
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    • pp.811-815
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    • 2005
  • The activated sludge from the aeration basin of the Su-yeong municipal wastewater treatment plant which has operated by a standard activated sludge process in Busan, Korea was investigated during April 2004 and January 2005 with several bio-indicators. The number of bacteria and fungi per gram of dry weight of MLSS were estimated to be $3.1\times10^6\sim1.5\times10^8\;and\;l.1\times10^3\sim1.1\times10^5$ colony forming units, respectively, by the plate agar method. By cultivation-independent methods, such as 4',6-diamidino-2-phenylindole stain and fluorescence in situ hybridization, the ratio of eubacteria to the entire biomass was evaluated by more than $80\%$ (v/v). The ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to the total eubacteria was detennined to be $7.0\sim9.8\%\;and\;3.3\sim6.2\%$ without heavy variation in spite of a period of relatively low temperature in the basin. It would be expected that the nitrification would occur or at least co-exist throughout the year in the sludge of many municipal WWTP with influents that contain the sufficient nitrogen sources although the WWTP does not have any specialized processes for the removal of nitrogen.

Cytogenetic Characteristics of Chinese Hamster Ovarian Cell CHO-K1

  • Sohn, Sea-Hwan;Cho, Eun-Jung;Jang, In-Surk
    • Reproductive and Developmental Biology
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    • v.30 no.4
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    • pp.263-270
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    • 2006
  • The Chinese Hamster Ovarian cells CHO-K1 are one of the most extensively used cells for the evaluation of gene expression and toxicology. However, these cells are frequently used for biomedical research without consideration of their cytogenetic characteristics. Therefore, we carried out to investigate the karyologic profiles, the frequency and type of chromosome aberration, and the distribution of telomeric DNA on chromosomes of the CHO-K1 cells. The GTG banding and fluorescence in situ hybridization on CHO-K1 cells were performed to characterize the karyotype and the distribution of telomeric DNA The present study revealed that the chromosome modal number of CHO-K1 cells was 2n=20; eight chromosomes appeared to be identical with those of the normal Chinese hamster, whereas the remaining 12 chromosomes were shown to be translocated, deleted, inversed, or rearranged from Chinese hamster chromosomes. The telomeric DNA on CHO-K1 chromosomes was intensively distributed at the centromeres rather than the ends of chromosomes. In addition, three chromosomes had interstitial telomeres and one marker chromosome entirely consisted of telomeric DNAs. The frequency and type of chromosome aberrations in CHO-K1 cells were examined. Of the 822 metaphase spreads, 68 (8.3%) cells resulted in chromosome aberrations of which the chromosome breakage was the most frequently occurred.

Karyotypic Analysis and Physical Mapping of rRNA Gene Loci in Persicaria tinctoria (쪽의 핵형분석과 rRNA 유전자의 염색체상 위치)

  • Choi, Hae-Woon;Lee, Sang-Hoon;Kim, Soo-Young;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.3
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    • pp.195-198
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    • 2008
  • Karyotypic analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rRNA genes were carried out in Persicaria tinctoria H Gross. The somatic metaphase chromosomes were ranged from 2.25 ${\mu}m$ to 1.50 ${\mu}m$ in length. Chromosome number was 2n = 4x = 40 with the basic number of x = 10. The chromosome complement of the species consisted of 16 pairs of metacentrics (chromososomes 1,2,3,4,6,7,8,9, 10, 11, 12, 13, 15, 18, 19 and 20) and 4 pairs of submetacentrics (chromosome 5, 14, 16 and 17). The karyotype formula was K(2n) = 4x = 32 m + 8 sm. In FISH analysis, three pairs of 45S rRNA gene loci on the terminal region of submetacentrics (chromosomes 5, 16 and 17) and two pairs of 5S rRNA gene loci on the centromeric region of metacentrics (chromosomes 9 and 11) were detected, respectively.

Cytogenetic Analysis Using Mitosis, Meiosis Chromosomes and bicolor Fluorescence in situ Hybridization of Bupleurum latissimum Nakai (체세포분열과 감수분열 및 bicolor FISH를 이용한 섬시호의 세포유전학적 분석)

  • Kim, Soo-Young;Bang, Jae-Wook;Lee, Joong-Ku
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.6
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    • pp.354-359
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    • 2006
  • Chromosome analysis using mitosis, meiosis and bicolor FISH were carried out in Bupleurum latissimum Nakai, which is one of the endemic plants in Ulleung island of korea. The somatic methaphase chromosomes number of this plant was 2n = 2x = 16 and the chromosome complements consisted of six pairs of metacentrics and two pairs of submetacentrics. The size of chromosomes ranged 2.40${\sim}$4.20 ${\mu}$m and NOR (nucleolus organizer region) chromosome did not observed using conventional staining. In meiosis chromosomes, metaphase-I and anaphase-I were observed. Metaphase-I anaphase-I showed 8 bivalents and chromosomes migration to make two daughter cells. Using bicolor FISH, one pair of 5S and 45S rDNA signals were detected on the centromeric region of chromosome 3 and the end of short of chromosome 2,respectively. We also observed the NOR using 45S rDNA probe.

A Case of Trisomy 22 with Pulmonary Hypoplasia in a Liveborn Infant (폐형성 부전이 동반된 Trisomy 22 1례)

  • Choi, Myoung-Bum;Kang, Ki Su;Park, Chan-Hoo;Woo, Hyang-Ok;Lee, Jeong-Hee;Lee, Jae-Ik;Paik, Won Young;Youn, Hee-Shang
    • Clinical and Experimental Pediatrics
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    • v.45 no.6
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    • pp.804-808
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    • 2002
  • We report a case of trisomy 22 in a liveborn male infant which was confirmed by fluorescence in situ hybridization(FISH), macrocultures and GTG-banding, and RHA-banding procedures of peripheral white blood cells. The infant showed lung hypoplasia, which is a unique presentation, with other clinical manifestations of previously reported cases of trisomy 22, such as intrauterine growth retardation, cleft palate, micrognathia, large atrial septal defect, limb anomalies, imperforate anus, and hypospadias. Our report gives weight to the previously reported observation that pulmonary hypoplasia may be associated in trisomy 22.

Physical Mapping of rDNAs Using McFISH in Anemarrhena asphodeloides Bunge (지모에서 McFISH를 이용한 rDNAs의 물리지도 작성)

  • Kim, Soo-Young;Choi, Hae-Woon;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.515-518
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    • 2004
  • Anemarrhena asphodeloides, a medicinal plant, has chromosome number of 2n=2x=22. To characterize the somatic metaphase chromosomes, physical mapping of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) was applied. Two pairs of 45S rDNA loci were detected on the terminal regions of the short arm of chromosomes 1 and 3. A pair of 5S rDNA signal was observed on the short arm of chromosome 3. 5S rDNA site seemed to be the same locus as one of the 45S rDNA site. McFISH was very useful tool for the localization and identification of rDNAs on the metaphase chromosomes in A. asphodeloides.

Study on Operational Factors in a Nitrite-Accumulating Submerged Membrane Bioreactor

  • Yoo Ik-Keun;Lim Kyoung-Jo;Lee Won-Sik;Kim Dong-Jin;Cha Gi-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.469-474
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    • 2006
  • Partial nitrification blocking of the oxidation of nitrite ($NO_{2}^{-}$) to nitrate ($NO_{3}^{-}$) has cost-efficient advantages such as lower oxygen and organics demand for nitrification and denitrification, respectively. A nitrifying membrane bioreactor of submerged type was operated for the treatment of synthetic ammonium wastewater with the purpose of nitrite build-up without affecting the efficiency of ammonium oxidation. A high ammonium concentration (1,000 mg/l) was completely converted to nitrate at up to 2 kg $N/m^3$ day under sufficient aeration. The control of pH under sufficient aeration was not a reliable strategy to maintain stable nitrite build-up. When the dissolved oxygen concentration was kept at 0.2-0.4 mg/l by adjusting the aeration rate, about 70% of nitrite content was obtained with ammonium oxidation efficiency higher than 93%. The increase of suction pressure due to membrane fouling was not significant under lowered aerating environment over a 6-month period of operation. The composition of nitrifier community, including relative abundance of nitrite oxidizers in a nitrite-accumulating condition, was quantified by fluorescence in situ hybridization analysis.

De novo interstitial deletion of 15q22q23 with global developmental delay and hypotonia: the first Korean case

  • Kim, Ha-Su;Han, Jin-Yeong;Kim, Myo-Jing
    • Clinical and Experimental Pediatrics
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    • v.58 no.8
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    • pp.313-316
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    • 2015
  • Interstitial deletions involving the chromosome band 15q22q24 are very rare and only nine cases have been previously reported. Here, we report on a 12-day-old patient with a de novo 15q22q23 interstitial deletion. He was born by elective cesarean section with a birth weight of 3,120 g at 41.3-week gestation. He presented with hypotonia, sensory and neural hearing loss, dysmorphism with frontal bossing, flat nasal bridge, microretrognathia with normal palate and uvula, thin upper lip in an inverted V-shape, a midline sacral dimple, severe calcanovalgus at admission, and severe global developmental delay at 18 months of age. Fluorescence in situ hybridization findings confirmed that the deleted regions contained at least 15q22. The chromosome analysis revealed a karyotype of 46,XY,del(15) (q22q23). Parental chromosome analysis was performed and results were normal. After reviewing the limited literature on interstitial 15q deletions, we believe that the presented case is the first description of mapping of an interstitial deletion involving the chromosome 15q22q23 segment in Korea. This report adds to the knowledge of the clinical phenotype associated with the 15q22q23 deletion.

An unusual de novo duplication 10p/deletion 10q syndrome: The first case in Korea

  • Lee, Bom-Yi;Park, Ju-Yeon;Lee, Yeon-Woo;Oh, Ah-Rum;Lee, Shin-Young;Choi, Eun-Young;Kim, Moon-Young;Ryu, Hyun-Mee;Park, So-Yeon
    • Journal of Genetic Medicine
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    • v.12 no.1
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    • pp.49-56
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    • 2015
  • We herein report an analysis of a female baby with a de novo dup(10p)/del(10q) chromosomal aberration. A prenatal cytogenetic analysis was performed owing to abnormal ultrasound findings including a choroid plexus cyst, prominent cisterna magna, and a slightly medially displaced stomach. The fetal karyotype showed additional material attached to the terminal region of chromosome 10q. Parental karyotypes were both normal. At birth, the baby showed hypotonia, upslanting palpebral fissures, a nodular back mass, respiratory distress, neonatal jaundice and a suspicious polycystic kidney. We ascertained that the karyotype of the baby was 46,XX,der(10)($pter{\rightarrow}q26.3::p11.2{\rightarrow}pter$) by cytogenetic and molecular cytogenetic analyses including high resolution GTG-and RBG-banding, fluorescence in situ hybridization, comparative genomic hybridization, and short tandem repeat marker analyses. While almost all reported cases of 10p duplication originated from one of the parents with a pericentric inversion, our case is extraordinarily rare as the de novo dup(10p)/del(10q) presumably originated from a rearrangement at the premeiotic stage of the parental germ cell or from parental germline mosaicism.