PMC7164144_01
Bone Marrow aspirate smears, Wright-Giemsa Stain 1000X
13039_2020_482_Fig1_HTML.jpg
PMC7164144_01_13039_2020_482_Fig1_HTML.jpg
Fig1
BM morphometric and flow cytometric analysis revealed proportionally normal myeloid, monocytic and lymphoid elements with no increased blasts, plasma cells and no aberrant myeloid or lymphoid maturation (Fig. 1). Bone marrow aspirate smears from the patient were stained with Wright- Giemsa Stain 1000X (Fig. 1).
G-banded metaphase chromosome preparations from unstimulated overnight bone marrow culture. All 20 chromosomes analyzed exhibited 44 chromosomes and no normal chromosome 14. Karyotype: 44,XY,rob(13;14) (q10,q10), rob(14;15) (q10;q10) [20] . The same heterologous RT formation was observed both in peripheral blood and epithelial tissue derived metaphase preparations. These karyotype results from multiple tissue types indicate the constitutional nature of the double RTs
13039_2020_482_Fig2_HTML.jpg
PMC7164144_01_13039_2020_482_Fig2_HTML.jpg
Fig2
FISH metaphase analysis proved the presence of 44 chromosomes containing (13;14) and (14;15) Robertsonian translocations (Figs. 2 and 4). Figure 2 shows the ISCN karyotype: 44,XY,rob(13;14)(q10;q10),rob(14;15)(q10;q10). No MDS/AML specific chromosome aberration was observed in the karyotype analysis or interphase and metaphase MDS FISH panel studies (Figs. 2 and 3). Cytomics FC500 with CXP software and fluorescent monoclonal antibodies (data not shown). Conventional G-banded metaphase chromosome analysis was performed on an unstimulated overnight bone marrow culture, a 72 h PHA-stimulated peripheral blood culture and bone marrow fibroblast cultures (Fig. 2).
FISH studies for MDS. BM metaphase spreads were hybridized with fluorescent DNA probes for: a: 5q31(EGR1 orange), 5p15.2(D5S721 green), b: 7q31(D7S486 orange), CEP 7(alpha satellite green) c: CEP 8(alpha satellite orange), d: 11q23(5'MLL green,3'MLL orange), e: 13q14.3(D13S319 orange), 13q34(green), f: 20q12 (D20S108 orange) No MDS specific FISH aberrations were observed
13039_2020_482_Fig3_HTML.jpg
PMC7164144_01_13039_2020_482_Fig3_HTML.jpg
Fig3
No MDS/AML specific chromosome aberration was observed in the karyotype analysis or interphase and metaphase MDS FISH panel studies (Figs. 2 and 3).
FISH characterization of double heterologous Robertsonian translocations. Bone marrow metaphase hybridized with fluorescent DNA probes for 15q22 (PML green), 15q11.2 (SNRPN orange), Cen15 (alpha satellite aqua), chromosome 14q32 (5'IGH green, 3'IGH orange) and chromosome 13q14(RB-1 green). Note the absence of a normal chromosome 14
13039_2020_482_Fig4_HTML.jpg
PMC7164144_01_13039_2020_482_Fig4_HTML.jpg
Fig4
FISH metaphase analysis proved the presence of 44 chromosomes containing (13;14) and (14;15) Robertsonian translocations (Figs. 2 and 4). Figure 2 shows the ISCN karyotype: 44,XY,rob(13;14)(q10;q10),rob(14;15)(q10;q10). A double heterozygous RT consisting of rob(13q;14q) and rob (14q;15q) was further confirmed by metaphase FISH analysis using multiple FISH probes encompassing various regions of chromosome 13, 14 and 15 (Fig. 4).