scholarly journals Analysis of Unrejoined Chromosomal Breakage in Human Fibroblast Cells Exposed to Low- and High-LET Radiation

2002 ◽  
Vol 43 (S) ◽  
pp. S181-S185 ◽  
Author(s):  
HONGLU WU ◽  
YOSHIYA FURUSAWA ◽  
KERRY GEORGE ◽  
TETSUYA KAWATA ◽  
FRANCIS A. CUCINOTTA
Keyword(s):  
Human Fibroblast ◽  
Fibroblast Cells ◽  
Chromosomal Breakage ◽  
Human Fibroblast Cells ◽  
High Let Radiation ◽  
High Let

G2 chromatid damage and repair kinetics in normal human fibroblast cells exposed to low- or high-LET radiation

2004 ◽  
Vol 104 (1-4) ◽  
pp. 211-215 ◽  
Author(s):  
T. Kawata ◽  
H. Ito ◽  
T. Uno ◽  
M. Saito ◽  
S. Yamamoto ◽  
...  
Keyword(s):  
Human Fibroblast ◽  
Fibroblast Cells ◽  
Normal Human Fibroblast ◽  
Human Fibroblast Cells ◽  
High Let Radiation ◽  
High Let ◽  
Normal Human

scholarly journals Induction of Chromatin Damage and Distribution of Isochromatid Breaks in Human Fibroblast Cells Exposed to Heavy Ions

2002 ◽  
Vol 43 (S) ◽  
pp. S169-S173 ◽  
Author(s):  
TETSUYA KAWATA ◽  
HISAO ITO ◽  
KEN MOTOORI ◽  
TAKUYA UEDA ◽  
NAOYUKI SHIGEMATSU ◽  
...  
Keyword(s):  
Human Fibroblast ◽  
Heavy Ions ◽  
Fibroblast Cells ◽  
Human Fibroblast Cells

scholarly journals Two Sp1/Sp3 Binding Sites in the Major Immediate-Early Proximal Enhancer of Human Cytomegalovirus Have a Significant Role in Viral Replication

2005 ◽  
Vol 79 (15) ◽  
pp. 9597-9607 ◽  
Author(s):  
Hiroki Isomura ◽  
Mark F. Stinski ◽  
Ayumi Kudoh ◽  
Tohru Daikoku ◽  
Noriko Shirata ◽  
...  
Keyword(s):  
Viral Replication ◽  
Human Cytomegalovirus ◽  
Significant Role ◽  
Binding Sites ◽  
Human Fibroblast ◽  
Immediate Early ◽  
Fibroblast Cells ◽  
Recombinant Viruses ◽  
Human Fibroblast Cells ◽  
Hcmv Replication

ABSTRACT We previously demonstrated that the major immediate early (MIE) proximal enhancer containing one GC box and the TATA box containing promoter are minimal elements required for transcription and viral replication in human fibroblast cells (H. Isomura, T. Tsurumi, M. F. Stinski, J. Virol. 78:12788-12799, 2004). After infection, the level of Sp1 increased while Sp3 remained constant. Here we report that either Sp1 or Sp3 transcription factors bind to the GC boxes located at approximately positions −55 and −75 relative to the transcription start site (+1). Both the Sp1 and Sp3 binding sites have a positive and synergistic effect on the human cytomegalovirus (HCMV) major immediate-early (MIE) promoter. There was little to no change in MIE transcription or viral replication for recombinant viruses with one or the other Sp1 or Sp3 binding site mutated. In contrast, mutation of both the Sp1 and Sp3 binding sites caused inefficient MIE transcription and viral replication. These data indicate that the Sp1 and Sp3 binding sites have a significant role in HCMV replication in human fibroblast cells.

O-083 Non-invasive, label-free optical analysis to detect aneuploidy within the inner cell mass of the preimplantation embryo

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Y Tan ◽  
S B Mahbub ◽  
C A Campugan ◽  
J Campbell ◽  
A Habibalahi ◽  
...  
Keyword(s):  
Hyperspectral Imaging ◽  
Human Fibroblast ◽  
Preimplantation Embryo ◽  
Mouse Embryos ◽  
Treated Mouse ◽  
Fibroblast Cells ◽  
Label Free ◽  
Primary Human Fibroblast ◽  
Non Invasive ◽  
Human Fibroblast Cells

Abstract Study question Can we separate between control and reversine-treated cells within the inner cell mass (ICM) of the mouse preimplantation embryo by using label-free and non-invasive hyperspectral microscopy? Summary answer Hyperspectral microscopy is able to discern between control and reversine-treated cells using cellular autofluorescence in the complete absence of fluorescence tags. What is known already Embryo mosaicism (containing cells that are euploid (46 chromosomes) and aneuploid (deviation from the expected number of chromosomes)) affects up to 17.3% of human blastocyst embryos. Current diagnosis of aneuploidy in the IVF clinic involves a biopsy of trophectoderm (TE) cells or spent media followed by sequencing. In some blastocyst embryos these approaches will fail to diagnose of the proportion of aneuploid cells within the fetal lineage (ICM). Study design, size, duration The impact of aneuploidy on cellular metabolism was assessed by using cellular autofluoresence and hyperspectral microscopy (broad spectral profile). Two models were employed: (i) Primary human fibroblast cells with known karyotypes (4-6 independent replicates, euploid n = 467; aneuploid n = 969) and reversine induced aneuploidy in mouse embryos (5-8 independent replicates, 30-44 cells per group). Both models were subjected to hyperspectral imaging to quantify native cell fluorescence. Participants/materials, setting, methods The human model is comprised of euploid (male and female) and aneuploid (triploid and trisomies: 13, 18, 21, XXX, and XXY) primary human fibroblast cells. For the mouse model, we treated embryos with reversine, a reversible spindle assembly checkpoint inhibitor, during the 4- to 8-cell division. Individual blastomeres were dissociated from control and reversine treated 8-cell embryos. Blastomeres were either imaged directly or used to generate chimeric blastocysts with differing ratios of control:reversine-treated cells. Main results and the role of chance Following unsupervised linear unmixing, the relative abundance of metabolic cofactors was quantified: reduced nicotinamide adenine dinucleotide (NAD(P)H) and flavins with the subsequent calculation of the optical redox ratio (ORR: Flavins/[NAD(P)H + Flavins]). Primary human fibroblast cells displayed an increase in the relative abundance of NAD(P)H with a decrease in flavins, leading to a significant reduction in the ORR for aneuploid cells (P < 0.05). The mouse embryos displayed an identical trend as the human model between control and reversine-treated embryos. Mathematical algorithms were applied and able to distinguish between (i) euploid and aneuploid primary human fibroblast cells, (ii) control and reversine-treated mouse blastomeres and (iii) chimeric blastocysts with differing ratios of control and reversine-treated cells. The accuracy of these separations was supported by receiver operating characteristic curves with areas under the curve. We also showed that hyperspectral imaging of the preimplantation embryo does not impact on embryo developmental competence, pregnancy outcome and offspring health in a mouse model. We believe the role of chance is low as both human somatic cells and mouse embryos showed a consistent shift in cellular metabolism in response to human fibroblast cells that are aneuploid and reversine treated mouse embryos. Limitations, reasons for caution Further validation of our approach could include sequencing of the ICM of individual blastocysts to determine the proportion of aneuploid cells in ICM and correlate this with the metabolic profile obtained through hyperspectral imaging. Wider implications of the findings With hyperspectral imaging able to discriminate between (i) euploid and aneuploid human fibroblast cells and (ii) control and reversine-treated mouse embryos, this could be an accurate, non-invasive and label-free optical imaging approach to assess mosaicism within the ICM of mouse embryos, potentially leading to a new diagnostic tool for embryos. Trial registration number Not applicable

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