• Title/Summary/Keyword: Fluorescent in situ hybridization

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Combined Study of Cytogenetics and Fluorescence in Situ Hybridization (FISH) Analysis in Childhood Acute Lymphoblastic Leukemia (ALL) in a Tertiary Cancer Centre in South India

  • Mazloumi, Seyed Hashem Mir;Madhumathi, D.S.;Appaji, L.;Prasannakumari, Prasannakumari
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3825-3827
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    • 2012
  • FISH is one of the most sensitive molecular methods to detect genetic abnormalities with DNA probes. When cytogenetic studies are normal or insufficient, FISH may detect cryptic rearrangements, rare or slowly proliferative abnormal populations in non-mitotic cells. We cytogenetically evaluated 70 childhood ALL - 67.1% were found to have an abnormal karyotype. The 23 patients (32.9%) with a normal karyotype were analyzed by FISH applying two probes; TEL/AML1 and MYB which detect cryptic rearrangements of t(12;21)(p13;q22) and deletion of (6q) respectively, associated with a good prognosis. Out of 23 patients, one was positive for t(12;21)(p13;q22) (4.3%). None of our patients were positive for MYB del(6q). Two patients showed an extra signal for MYB on chromosomes other than 6 (8.6 %) indicating amplification or duplication. Findings were compared with the available literature. Our study clearly indicated the integrated FISH screening method to increase the abnormality detection rate in a narrow range. FISH is less useful for diagnostic study of patients with suspected del(6q) but it helps in detecting known cryptic rearrangements as well as identification of new abnormalities(translocation , duplication and amplification) at the gene level.

Monitoring of Bacterial Community in a Coniferous Forest Soil After a Wildfire

  • Kim Ok-Sun;Yoo Jae-Jun;Lee Dong-Hun;Ahn Tae-Seok;Song Hong-Gyu
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.278-284
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    • 2004
  • Changes in the soil bacterial community of a coniferous forest were analyzed to assess microbial responses to wildfire. Soil samples were collected from three different depths in lightly and severely burned areas, as well as a nearby unburned control area. Direct bacterial counts ranged from $3.3­22.6\times10^8\;cells/(g{\cdot}soil).$ In surface soil, direct bacterial counts of unburned soil exhibited a great degree of fluctuation. Those in lightly burned soil changed less, but no significant variation was observed in the severely burned soil. The fluctuations of direct bacterial count were less in the middle and deep soil lay­ers. The structure of the bacterial community was analyzed via the fluorescent in situ hybridization method. The number of bacteria detected with the eubacteria-targeted probe out of the direct bacterial count varied from $30.3\;to\;84.7\%,$ and these ratios were generally higher in the burned soils than in the unburned control soils. In the surface unburned soil, the ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ Cytoph­aga-Flavobacterium group, and other eubacteria groups to total eubacteria were 9.9, 10.6, 15.5, 9.0, and $55.0\%,$ respectively, and these ratios were relatively stable. The ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ and Cytophaga-Flavobacterium group to total eubacteria increased immediately after the wildfire, and the other eubacterial proportions decreased in the surface and middle layer soils. By way of contrast, the composition of the 5 groups of eubacteria in the subsurface soil exhibited no significant fluctuations dur­ing the entire period. The total bacterial population and bacterial community structure disturbed by wildfire soon began to recover, and original levels seemed to be restored 3 months after the wildfire.

Community Analysis of the Bacteria in Sponges of Lake Baikal by FISH Method (바이칼 호 Sponge에 서식하는 세균 군집 구조의 분석)

  • Seo, Eun-Young;Kim, Mi-Ree;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.14-18
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    • 2007
  • The bacterial community structures at 2 sponge species belonging to the genus Baikalospongia and Lubomirskia in Lake Baikal were analyzed with fluorescent in situ hybridization (FISH) method. The total bacterial numbers in the genus Baikalospongia ranged from $7.2{\times}10^{7}\;to\;4.2{\times}10^{8}\;cells/ml$, and those in Lubomirskia from $1.2{\times}10^{8}\;to\;1.6{\times}10^{8}\;cells/ml$, while those of lake water were from $2.3{\times}7.7{\times}10^{5}\;cells/ml$. Total bacterial numbers in the sponges were $10^{3}-10^{4}$ times higher than those of lake water. In the genera Baikalospongia and Lubomirskia, the proportions of other unidentified bacterial groups to the Bacteria were 42.0-60.3% and 40.7-51.9%, respectively. These proportions were similar to those in lake water (22.6-46.3%). These results suggest that bacterial compositions in Lake Baikal water and sponges are highly unique.

Prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus

  • Lee, Dongsook;Park, Heeju;Kwak, Sanha;Lee, Soomin;Go, Sanghee;Park, Sohyun;Jo, Sukyung;Kim, Kichul;Lee, Seunggwan;Hwang, Doyeong
    • Journal of Genetic Medicine
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    • v.13 no.2
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    • pp.95-98
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    • 2016
  • We report the prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus. Cytogenetic analysis of parental chromosomes revealed that the mother had a normal 46,XX karyotype, whereas the father exhibited a 46,XY,der(15)t(Y;15) karyotype. We performed cytogenetic analysis of the father's family as a result of the father and confirmed the same karyotype in his mother and brother. Fluorescence in situ hybridization and quantitative fluorescent-polymerase chain reaction analysis identified the breakpoint and demonstrated the absence of the SRY gene in female members. Thus, the proband inherited this translocation from the father and grandmother. This makes the prediction of the fetal phenotype possible through assessing the grandmother. Therefore, we suggest that conventional cytogenetic and molecular cytogenetic methods, in combination with family history, provide informative results for prenatal diagnosis and prenatal genetic counseling.

Correlationship of Vertical Distribution for Ammonia Ion, Nitrate Ion and Nitrifying Bacteria in a Fixed Bed Nitrifying Biofilm

  • Choi, Gi-Chung;Byun, Im-Gyu
    • Journal of Environmental Science International
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    • v.21 no.12
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    • pp.1455-1462
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    • 2012
  • The vertical distributions of nitrifying bacteria in aerobic fixed biofilm were investigated to evaluate the relationship between nitrification performance and microbial community at different HRT. Fluorescent in situ hybridization (FISH) and portable ion selective microelectrode system were adopted to analyze microbial communities and ions profiles according to the biofilm depth. Cilia media packed MLE (Modified Ludzack-Ettinger) like reactor composed of anoxic, aerobic I/II was operated with synthetic wastewater having COD 200 mg/L and $NH_4{^+}$-N mg/L at HRT of 6 hrs and 4 hrs. Total biofilm thickness of aerobic I, II reactor at 4 hrs condition was over two times than that of 6 hrs condition due to the sufficient substrate supply at 4 hrs condition (6 hrs; aerobic I 380 ${\mu}m$ and II 400 ${\mu}m$, 4 hrs; aerobic I 830 ${\mu}m$ and II 1040 ${\mu}m$). As deepen the biofilm detection point, the ratio of ammonia oxidizing bacteria (AOB) was decreased while the ratio of nitrite oxidizing bacteria (NOB) was maintained similar distribution at both HRT condition. The ratio of AOB was higher at 4 hrs than 6 hrs condition and $NH_4{^+}$-N removal efficiency was also higher at 4 hrs with 89.2% than 65.4% of 6 hrs. However, the ratio of NOB was decreased when HRT was reduced from 6 hrs to 4 hrs and $NO_2{^-}$-N accumulation was observed at 4 hrs condition. Therefore, it is considered that insufficient HRT condition could supply sufficient substrate and enrichment of AOB in all depth of fixed biofilm but cause decrease of NOB and nitrite accumulation.

The Bacterial Community Structure in Cheonho Reservoir Dominated by Cyanobacteria (봄철 Cyanobacteria 가 우점한 천호지에서 세균군집구조의 변화)

  • 홍선희;전선옥;안태석;안태영
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.287-292
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    • 2002
  • The composition of bacterial communities was detected in surface water of Cheonho Reservoir dominated by cyanobacteria, using fluorescent in situ hybridization (FISH) method. Total bacterial numbers were very high ranging from 0.6~$1.3{\times}10^7 \cells{\cdot}ml^-1$, whereas the ratio of Eubacteria to total bacteria was 29.8~45.8%, which was lower than that in other freshwater ecosystems. On average only 2.1% of DAPI-stained bacteria were detected by FISH with probes for $\alpha$, $\beta$, and $\gamma$-groups, respectively. Unknown eubacteria which was not bound to any probes except EUB 338, was relatively high. On the other hand, the Cytophaga-Flavobacterium group increased following the change of dominant species from Anabaena sp. to Microcystis sp. This result showed that bacterial communities could be affected by phytoplanktons, especially cyanobacteria.

Development of FISH Methods for Detection of Genus Bacillus (Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.204-208
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    • 2001
  • A technique for detection of Bacillus in soil and waste water treatment system was developed. Mixed col- tured solutions of 22 Bacillus strains were applied for selection of probe and pretreatment method for FISHAmong the three probes known as useful tool for FISH method,S-G-Bacill-0597-a-A-22 was best for detec-tion of Bacillus. Ninety five percent of DAPl count was observed with FISH method with S-G-Bacill-0597-a-A-22 probe. For increasing the permeability to Bacillus cell walls, pretreatment with lysozyme was betterthan that with lysozyme and SDS/DTT (dithiothreitol). Bacillus spore was not detected with FISH. So, Bacil-lus detection in ecosystem requires FISH with pretreatment of lysozyme and spore staining.

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THE EFFECT OF XYLITOL ON THE EXPRESSION OF GTF GENE (gtf 유전자 발현에 대한 xylitol의 영향)

  • Yeom, Chung-Hyun;Chung, Jin;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.2
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    • pp.304-313
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    • 2004
  • Xylitol, a five-carbon natural sugar alcohol, is widely used non-cariogenic sugar substitute. In present study, the effects of xylitol on the expression of mRNA for glucosyltransferase which synthesizes glucan from sucrose were detected by Fluorescent in situ hybridization (FISH) and flow cytometry. FITC fluorescences for mRNA of gtfB, gtfC and gtfD were decreased further with increasing concentration of xylitol from 1% to 10% when detected by FISH. Flow cytometric analysis also showed that the expression of gtfB, gtfC and gtfD was increased by the addition of sucrose and decreased by the addition of xylitol to BHI broth containing 1% sucrose. In conclusion, the expression of gtfB, gtfC and gtfD mRNA was decreased by the addition of xylitol.

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Seasonal and Vertical Change of Bacterial Communities in Lake Soyang (소양호에서 세균군집구조의 계절적.수직적 변화)

  • 김동주;홍선희;안태석
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.242-247
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    • 1999
  • This sludy was conducted to investigate the change of bacterial co~munities with season and depth in Lake Soyang. Korea, using fluorescence in situ hybridization (FISH). The oligonucleotide probes used in this study were EUB338, ALF I b. GAM42a, and CF, The percentage of h e Proleobacteria a $\alpha$-group ranged from 0.70 to 33% the $\beta$-group from 1.0 to 26% they -group from 2.4 Lo 37% and Cytophagn and Flavobactefin groups from 4.7 to 24% duing the study period (April Lo November, 1998). They $\gamma$-group was dominant in spring when Asterionella was dominant. and a $\alpha$-group was dominant in summer when the organic content was low and Dinobryon was dominant. However, a specific group was not dominant in ?dl when cyanobacteria group was dominant and the ratio of eubacleria to total bacteria was very low. Therefore, the bacterial communities in Lake Soyang changed with season and depth, which seems to be associated with the telnporal succession of phytoplanlaons.

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Dynamics of in situ Bacterial Community Structure in the Nak-Dong River (낙동강에서의 세균군집구조의 역동성)

  • Park, Ji-Eun;Yeo, Sang-Min;Lee, Young-Ok
    • Korean Journal of Ecology and Environment
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    • v.37 no.4 s.109
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    • pp.363-367
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    • 2004
  • For comparative analysis of the eubacterial community structure at 8 sampling sites throughout the Nak-Dong River, FISH (fluorescence in situ hybridization) method was employed. The total ratio of each determined eubacterial group such as ${\alpha}\;{\cdot}\;{\beta}\;{\cdot}\;{\gamma}-subclasses$proteobacteria and Cytophaga-Flavobacterium(CF) group to total counts(DAPI) at each site varied 9.3-42.5% with the highest value at uppermost part. And each ratio of determined eubacterial groups reached mostly under 10% except that of CF group (23%) at uppermost part. Furthermore, compared to lower part, upper part represented unexpectedly higher proportions of ${\gamma}-subclass$ proteobacteria comprised almost fast growing bacteria on degradable organics. Also the variations of ammonia-oxidizing bacteria ranged from $2.7{\times}10^4$ to $18.0{\times}10^4$ cells $mL^{-1}$ with the lowest value in lower part and the highest value in mid part whereas those of nitrite-oxidizing bacteria varied 5.2-7.7{\times}10^4$ cells $mL^{-1}$ without noticeable differences throughout the sites. Additionally, the ratio of nitrifying bacteria to total counts ranged from 1.0% to 13.6% with no differences between ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. In conclusion, FISH method introduced in this study for monitoring, normally used for the quantitative analysis of bacteria, provided also good information on their environmental status in the Nak-Dong River.