Could Cobalamin Deficiency Mimic a Clonal Aberration Cytogenetic? a Case Report

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4882-4882
Author(s):  
Marcelo Bellesso ◽  
Daniela Ferreira Dias ◽  
Renato Torrescasana Centrone ◽  
Laura Yolanda Chialanza Garcia ◽  
Annelise Correa Wengerkievicz Lopes ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Vitamin B12 ◽  
Cytogenetic Analysis ◽  
Bone Marrow Aspirate ◽  
Bone Marrow Cells ◽  
Megaloblastic Anemia ◽  
Cobalamin Deficiency ◽  
Normal Range ◽  
Clonal Deletion ◽  
Marrow Cells

Abstract A 32 years old female patient presented to our service complaining asthenia, apathy and loss of her power to work. It was observed, by laboratory test, pancytopenia (Hb: 6.0 g/dL; WBC: 2,100/mm³, platelets 95,000/mm³, Reticulocytes 0.8%) associated with marked elevation of LDH 1,800 U/L (normal range: 240 – 480U/L). Bone marrow aspirate demonstrated morphologic features of megaloblastic anemia. Moreover, low serum concentration of vitamin B12 < 150pg/mL (normal range: 200 – 95-pg/mL), Folic acid 15,9ng/mL (normal range: 3 – 17 ng/mL) confirmed Megaloblastic anemia by cobalamin deficiency. It was not evidenced gastritis. It was initiated the treatment with vitamin B12. However, in the next clinical attendance it was observed an unexpected cytogenetic result: 46,XX,del(9)(q13q22)[3]/76~78,XXX,+1,+3,+4,+9,+11,+12,+17,+20,+21[cp3]/46,XX[34]. It was interpreted that del(9)(q13q22)[3] could be a clonal cytogenetics aberration and the others data due to defects in synthesis of DNA by cobalamin deficiency. It was important, because in diagnosis of Megaloblastic anemia frequently it is not included cytogenetic analysis of bone marrow cells and a new cytogenetic analysis has become necessary due to this data. After two months, a complete hematological recovery was achieved and six months later, bone marrow aspirate and cytogenetic analisys were repeated. Normal morphologic bone marrow cells and normal cytogenetic: 46,XX[20] were evidenced. Therefore, it was really hard to conclude this case. First, cobalamin deficiency could promote this clonal deletion or, as a second hypothesis, a clonal cytogenetic with low proliferative rate was selected due to inefficient hematopoiesis, by vitamin B12 deficiency, and after the recovery of the hematopoiesis, this clonal was suppressed. Disclosures No relevant conflicts of interest to declare.

scholarly journals Bone marrow cells from vitamin B12- and folate-deficient patients misincorporate uracil into DNA

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1656-1661 ◽  
Author(s):  
SN Wickramasinghe ◽  
S Fida
Keyword(s):  
Bone Marrow ◽  
Vitamin B12 ◽  
Bone Marrow Cells ◽  
Megaloblastic Anemia ◽  
Folate Deficiency ◽  
Test Results ◽  
Dnase 1 ◽  
Biochemical Lesion ◽  
Suppression Test ◽  
Marrow Cells

Bone marrow cells from 15 patients with normal deoxyuridine (dU) suppression test results, 3 healthy subjects, and 11 patients with megaloblastic anemia caused by vitamin B12 or folate deficiency were examined for misincorporation of uracil into DNA. Cells were incubated with [5–3H] uridine for 2 hours and their DNA extracted. The DNA was hydrolyzed to deoxyribonucleosides with DNase 1, phosphodiesterase and alkaline phosphatase, and any dU present was separated from other deoxyribonucleosides by Aminex A6 chromatography. The quantity of dU/mg DNA and the radioactivity in the dU peak/mg DNA were then calculated. The results clearly showed that there was markedly increased uracil misincorporation into the DNA of vitamin B12- or folate-deficient marrow cells. Misincorporation of uracil into DNA may be an important biochemical lesion underlying both the megaloblastic change and the ineffectiveness of hematopoiesis in vitamin B12 and folate deficiency.

scholarly journals Bone marrow cells from vitamin B12- and folate-deficient patients misincorporate uracil into DNA

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1656-1661 ◽  
Author(s):  
SN Wickramasinghe ◽  
S Fida
Keyword(s):  
Bone Marrow ◽  
Vitamin B12 ◽  
Bone Marrow Cells ◽  
Megaloblastic Anemia ◽  
Folate Deficiency ◽  
Test Results ◽  
Dnase 1 ◽  
Biochemical Lesion ◽  
Suppression Test ◽  
Marrow Cells

Abstract Bone marrow cells from 15 patients with normal deoxyuridine (dU) suppression test results, 3 healthy subjects, and 11 patients with megaloblastic anemia caused by vitamin B12 or folate deficiency were examined for misincorporation of uracil into DNA. Cells were incubated with [5–3H] uridine for 2 hours and their DNA extracted. The DNA was hydrolyzed to deoxyribonucleosides with DNase 1, phosphodiesterase and alkaline phosphatase, and any dU present was separated from other deoxyribonucleosides by Aminex A6 chromatography. The quantity of dU/mg DNA and the radioactivity in the dU peak/mg DNA were then calculated. The results clearly showed that there was markedly increased uracil misincorporation into the DNA of vitamin B12- or folate-deficient marrow cells. Misincorporation of uracil into DNA may be an important biochemical lesion underlying both the megaloblastic change and the ineffectiveness of hematopoiesis in vitamin B12 and folate deficiency.

scholarly journals 5-Methyltetrahydrofolate related enzymes and DNA polymerase alpha activities in bone marrow cells from patients with vitamin B12 deficient megaloblastic anemia

Blood ◽  
1982 ◽  
Vol 59 (4) ◽  
pp. 832-837
Author(s):  
Y Kano ◽  
S Sakamoto ◽  
K Hida ◽  
K Suda ◽  
F Takaku
Keyword(s):  
Bone Marrow ◽  
Vitamin B12 ◽  
Dna Polymerase ◽  
Methylenetetrahydrofolate Reductase ◽  
Bone Marrow Cells ◽  
Megaloblastic Anemia ◽  
Vitamin B12 Deficiency ◽  
Dna Polymerase Alpha ◽  
B12 Deficiency ◽  
Marrow Cells

The activities of 5-methyltetrahydrofolate (5-CH3THF) related enzymes and DNA polymerase alpha were determined in bone marrow cells obtained from patients with vitamin B12 deficient megaloblastic anemia and compared with those from healthy volunteers and patients with hemolytic anemia. 5-CH3THF homocysteine methyltransferase activity was significantly lower than that in the control subjects. 5,10- methylenetetrahydrofolate reductase activity was only slightly elevated to that in the control subjects. DNA polymerase alpha activity was significantly higher than that in the control. High deoxyuridine suppression test values in vitamin B12 deficient bone marrow cells were improved by tetrahydrofolate, but not by 5-CH3THF. These data indicate that, even though the reverse reaction catalyzed by 5,10- methylenetetrahydrofolate reductase may be operative in vitamin B12 deficiency, it is not sufficient to correct the disturbance in folate metabolism in vitamin B12 deficiency. Increased DNA polymerase alpha activity may be due to compensation for disarranged DNA synthesis.

scholarly journals 5-Methyltetrahydrofolate related enzymes and DNA polymerase alpha activities in bone marrow cells from patients with vitamin B12 deficient megaloblastic anemia

Blood ◽  
1982 ◽  
Vol 59 (4) ◽  
pp. 832-837 ◽  
Author(s):  
Y Kano ◽  
S Sakamoto ◽  
K Hida ◽  
K Suda ◽  
F Takaku
Keyword(s):  
Bone Marrow ◽  
Vitamin B12 ◽  
Dna Polymerase ◽  
Methylenetetrahydrofolate Reductase ◽  
Bone Marrow Cells ◽  
Megaloblastic Anemia ◽  
Vitamin B12 Deficiency ◽  
Dna Polymerase Alpha ◽  
B12 Deficiency ◽  
Marrow Cells

Abstract The activities of 5-methyltetrahydrofolate (5-CH3THF) related enzymes and DNA polymerase alpha were determined in bone marrow cells obtained from patients with vitamin B12 deficient megaloblastic anemia and compared with those from healthy volunteers and patients with hemolytic anemia. 5-CH3THF homocysteine methyltransferase activity was significantly lower than that in the control subjects. 5,10- methylenetetrahydrofolate reductase activity was only slightly elevated to that in the control subjects. DNA polymerase alpha activity was significantly higher than that in the control. High deoxyuridine suppression test values in vitamin B12 deficient bone marrow cells were improved by tetrahydrofolate, but not by 5-CH3THF. These data indicate that, even though the reverse reaction catalyzed by 5,10- methylenetetrahydrofolate reductase may be operative in vitamin B12 deficiency, it is not sufficient to correct the disturbance in folate metabolism in vitamin B12 deficiency. Increased DNA polymerase alpha activity may be due to compensation for disarranged DNA synthesis.

scholarly journals Clonal deletion of postthymic T cells: veto cells kill precursor cytotoxic T lymphocytes.

1992 ◽  
Vol 175 (3) ◽  
pp. 863-868 ◽  
Author(s):  
K Hiruma ◽  
H Nakamura ◽  
P A Henkart ◽  
R E Gress
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Bone Marrow Cells ◽  
High Dose ◽  
Spleen Cells ◽  
Clonal Deletion ◽  
Veto Cells ◽  
Veto Cell ◽  
Marrow Cells ◽  
Ctl Responses

Veto cell-mediated suppression of cytotoxic T lymphocyte (CTL) responses has been proposed as one mechanism by which self-tolerance is maintained in mature T cell populations. We have previously reported that murine bone marrow cells cultured in the presence of high-dose interleukin 2 (IL-2) (activated bone marrow cells [ABM]) mediate strong veto suppressor function. To examine mechanisms by which ABM may suppress precursor CTL (p-CTL) responses, we used p-CTL generated from spleen cells of transgenic mice expressing a T cell receptor specific for H-2 Ld. It was demonstrated that the cytotoxic response by these p-CTL after stimulation with irradiated H-2d/k spleen cells was suppressed by DBA/2 (H-2d) ABM, but not by B10.BR (H-2k) ABM or dm1 (Dd, Ld mutant) ABM. Flow cytometry analysis with propidium iodide staining revealed that these p-CTL were specifically deleted by incubation with H-2d ABM, but not with H-2k ABM. These data indicate that ABM veto cells kill p-CTL with specificity for antigens expressed on the surface of the ABM, and that the mechanism for veto cell activity of ABM is clonal deletion of p-CTL.

scholarly journals Cytogenetic analysis of bone marrow cells of rats treated with toluene

2004 ◽  
Vol 58 (3-4) ◽  
pp. 311-318
Author(s):  
Svetlana Fister ◽  
Slavoljub Jovic ◽  
Jelka Stevanovic ◽  
Milica Kovacevic-Filipovic
Keyword(s):  
Bone Marrow ◽  
Mitotic Index ◽  
Cytogenetic Analysis ◽  
Wistar Rats ◽  
Bone Marrow Cells ◽  
Experimental Conditions ◽  
Structural Aberrations ◽  
Female Wistar Rats ◽  
Cytogenetic Analyses ◽  
Marrow Cells

The paper presents the results of investigations of the effect of toluene on bone marrow cells of female Wistar rats treated intraperitoneally with toluene for 8 or 11 days, in doses of 0.602 ?g/200 g body mass. Cytogenetic analyses were performed on the metaphase figure of chromosomes in order to determine the frequency of structural aberrations ? breaks and gaps. The values of the mitotic index and number of poliploid cells were determined. No significant increase was determined in the frequency of breaks and gaps in chromosomes of treated animals in comparison with the controls, which means that, under the experimental conditions, toluene did not exhibit a definite genotoxic effect. However, it has been determined that there was a significant increase in the value of the mitotic index, as well as a significant increase in the number of poliploid cells in both groups of treated rats in comparison with controls.

scholarly journals Cellular basis of graft versus host tolerance in chimeras prepared with total lymphoid irradiation.

1980 ◽  
Vol 152 (3) ◽  
pp. 736-741 ◽  
Author(s):  
M Gottlieb ◽  
S Strober ◽  
H S Kaplan
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Suppressor Cells ◽  
Third Party ◽  
Whole Body ◽  
Mixed Chimerism ◽  
Clonal Deletion ◽  
Donor Type ◽  
Host Tolerance ◽  
Marrow Cells

BALB/c mice given allogeneic (C57BL/Ka) bone marrow cells after toal lymphoid irradiation become stable chimeras approximately 80% donor-type and 20% host-type cells in the spleen. The chimeras doe not develop graft vs. host disease (GVHD). Purified cells of C57BL/Ka origin from the chimeras mediated GVHD in lightly irradiated C3H (third party), but not in BALB/c (host-strain) mice. Thus graft vs. host tolerance in the chimeras could not be explained by complete immunodeficiency of donor-type cells, serum blocking factors, or suppressor cells of host (BALB/c) origin. Clonal deletion or suppression of lymphocytes reactive with host tissues remain possible explanations. The transfer of donor-type chimeric spleen cells to BALB/c recipients given 500-550 rad whole-body irradiation WBI led to stable mixed chimerism in approximately 50% of recipients. The cells were presumably acting as tolerogens because similarly irradiated BALB/c mice given (BALB/c X C57BL/Ka)F1 spleen or bone marrow cells also became stable mixed chimeras.

scholarly journals Stimulation of proliferation of human myeloid leukemia cells in culture: applications for cytogenetic analysis

Blood ◽  
1986 ◽  
Vol 68 (3) ◽  
pp. 790-793
Author(s):  
J Michaeli ◽  
I Lerer ◽  
EA Rachmilewitz ◽  
E Fibach
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Peripheral Blood ◽  
Cytogenetic Analysis ◽  
Bone Marrow Cells ◽  
Leukemic Cells ◽  
Pronounced Increase ◽  
Stimulation Of ◽  
Marrow Cells ◽  
Mitotic Cells

The use of chromosome banding techniques has provided a valuable diagnostic tool in various malignancies. The application of these methods, however, is often restricted by a low yield of mitotic cells and the patient's unwillingness to comply with repeated bone marrow aspiration. In an attempt to promote mitotic activity of leukemic cells from the bone marrow and peripheral blood, we employed a new method based on culturing the cells in the presence of a conditioned medium derived from a human bladder carcinoma cell line (5637). In addition to colony stimulating factor, this conditioned medium contains a factor that is capable of stimulating leukemic myeloblast proliferation. Bone marrow and peripheral blood mononuclear cells from 58 patients with a variety of myeloid leukemias were cultured for 24 to 120 hours in the presence or absence of conditioned medium. These bone marrow cells showed a pronounced increase in the mitotic index (5- to 50-fold) as compared to unstimulated cultures, and a greater than 100-fold increase as compared to fresh, uncultured bone marrow cells. Analyzable metaphases could be obtained even in marrow samples in which direct or 24-hour G-banding techniques had failed to reveal metaphases. The effect observed on peripheral blood cells was even more dramatic because prior to culture no mitotic cells were detected, whereas up to 2% mitotic cells were found in conditioned medium-stimulated peripheral leukemic cells. Karyotype analysis of 36 out of the 58 leukemic patients has shown that the chromosome changes discovered in conditioned medium stimulated cells were identical to those found in unstimulated cells. New chromosome aberrations, attributable to the stimulation of growth by conditioned medium, were not found. The quality of the metaphases analyzed following conditioned medium stimulation was considerably better than that of unstimulated samples. Frozen cells, when cultured with conditioned medium, were also suitable for cytogenetic analysis. Thus, the use of this conditioned medium permits adequate cytogenetic analysis even in cases where such analysis was previously impossible.

scholarly journals Defect in negative selection in lpr donor-derived T cells differentiating in non-lpr host thymus.

1991 ◽  
Vol 173 (1) ◽  
pp. 127-136 ◽  
Author(s):  
K Matsumoto ◽  
Y Yoshikai ◽  
T Asano ◽  
K Himeno ◽  
A Iwasaki ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Cells ◽  
Interleukin 2 ◽  
Receptor Expression ◽  
Donor Strain ◽  
Clonal Deletion ◽  
Massive Lymphadenopathy ◽  
Single Positive ◽  
Marrow Cells

Transplantation of bone marrow cells of lpr/lpr mice into irradiated normal mice fails to develop massive lymphadenopathy or autoimmunity but causes severe graft-vs.-host-like syndrome. To elucidate an abnormality of lpr/lpr bone marrow-derived T cells, we transplanted bone marrow cells of Mlsb lpr/lpr mice into H-2-compatible Mlsa non-lpr mice. Although lpr/lpr T cell precursors repopulated the host thymus as well as +/+ cells, a proportion of CD4+CD8+ cells decreased, and that of both CD4- and CD8- single-positive cells increased compared with those of +/+ recipients. Notably, in MRL/lpr----AKR and C3H/lpr----AKR chimeras, CD4 single-positive thymocytes contained an increased number of V beta 6+ cells in spite of potentially deleting alleles of Mlsa, whereas V beta 6+ mature T cells were deleted in the MRL/+ ----AKR and C3H/+ ----AKR chimeras. There was no difference between MRL/+ ----AKR and MRL/lpr----AKR chimeras in their proportion of V beta 3+ cells because both host and donor strain lack the deleting alleles. Interleukin 2 receptor expression of mature T cells, in the thymus and lymph node, was obviously higher in the MRL/lpr----AKR chimeras, in particular in the "forbidden" V beta 6+ subset. Moreover, lpr donor-derived peripheral T cells showed vigorous anti-CD3 response. These results indicate that lpr-derived T cells escape not only tolerance-related clonal deletion but also some induction of unresponsiveness in the non-lpr thymus.

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