scholarly journals Multiple Rh messenger RNA isoforms are produced by alternative splicing

Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1074-1078 ◽  
Author(s):  
C Le Van Kim ◽  
B Cherif-Zahar ◽  
V Raynal ◽  
I Mouro ◽  
M Lopez ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Messenger Rna ◽  
Cell Lineage ◽  
Pcr Amplification ◽  
Human Bone ◽  
Protein Isoforms ◽  
Human Bone Marrow ◽  
Chain Reaction ◽  
Splicing Isoforms ◽  
Polymerase Chain

Three Rh-related cDNAs have been isolated from a human bone marrow cDNA library and by polymerase chain reaction (PCR) amplification of human bone marrow and erythroblast mRNAs. They potentially encode a family of Rh protein isoforms that exhibit several unexpected structural properties as compared with the Rh polypeptide encoded by the cDNA clone identified previously. These modifications include several peptide deletions, the predicted alteration of Rh protein topology within the cell membrane, variations in the number and surface exposition of cysteine residues, and the generation of new C-terminal polypeptide segments caused by frameshift mutations. The four Rh mRNAs now described correspond to different splicing isoforms transcribed from the same Rh gene, and all exist in the same cell lineage (erythroid). Moreover, PCR experiments indicated that at least three of these RNA species exist in reticulocytes from donors with different commonly expressed Rh phenotypes. Although the translated proteins have not yet been characterized, these results suggest that the two genes at the RH locus may direct the synthesis of several protein species possibly corresponding to different Rh antigenic variants.

scholarly journals Multiple Rh messenger RNA isoforms are produced by alternative splicing

Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1074-1078 ◽  
Author(s):  
C Le Van Kim ◽  
B Cherif-Zahar ◽  
V Raynal ◽  
I Mouro ◽  
M Lopez ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Messenger Rna ◽  
Cell Lineage ◽  
Pcr Amplification ◽  
Human Bone ◽  
Protein Isoforms ◽  
Human Bone Marrow ◽  
Chain Reaction ◽  
Splicing Isoforms ◽  
Polymerase Chain

Abstract Three Rh-related cDNAs have been isolated from a human bone marrow cDNA library and by polymerase chain reaction (PCR) amplification of human bone marrow and erythroblast mRNAs. They potentially encode a family of Rh protein isoforms that exhibit several unexpected structural properties as compared with the Rh polypeptide encoded by the cDNA clone identified previously. These modifications include several peptide deletions, the predicted alteration of Rh protein topology within the cell membrane, variations in the number and surface exposition of cysteine residues, and the generation of new C-terminal polypeptide segments caused by frameshift mutations. The four Rh mRNAs now described correspond to different splicing isoforms transcribed from the same Rh gene, and all exist in the same cell lineage (erythroid). Moreover, PCR experiments indicated that at least three of these RNA species exist in reticulocytes from donors with different commonly expressed Rh phenotypes. Although the translated proteins have not yet been characterized, these results suggest that the two genes at the RH locus may direct the synthesis of several protein species possibly corresponding to different Rh antigenic variants.

The Effect of the MicroRNA-183 Family on Hair Cell-Specific Markers of Human Bone Marrow-Derived Mesenchymal Stem Cells

2018 ◽  
Vol 23 (4) ◽  
pp. 208-215 ◽  
Author(s):  
Mohammad-Reza  Mahmoudian-Sani ◽  
Mohammad-Saeid Jami ◽  
Ali Mahdavinezhad ◽  
Razieh Amini ◽  
Gholamreza  Farnoosh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Mesenchymal Stem Cells ◽  
Hair Cell ◽  
Hair Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Chain Reaction ◽  
Polymerase Chain

Hearing loss is considered the most common sensory disorder across the world. Nowadays, a cochlear implant can be an effective treatment for patients. Moreover, it is often believed that sensorineural hearing loss in humans is caused by loss or disruption of the function of hair cells in the cochlea. In this respect, mesenchymal cells can be a good candidate for cell-based therapeutic approaches. To this end, the potential of human bone marrow-derived mesenchymal stem cells to differentiate into hair cells with the help of transfection of microRNA in vitro was investigated. MicroRNA mimics (miRNA-96, 182, and 183) were transfected to human bone marrow-derived mesenchymal stem cells using Lipofec­tamine as a common transfection reagent following the manufacturer’s instructions at 50 nM for microRNA mimics and 50 nM for the scramble. The changes in cell morphology were also observed under an inverted microscope. Then, the relative expression levels of SOX2, POU4F3, MYO7A, and calretinin were assayed using real-time polymerase chain reaction according to the ΔΔCt method. The ATOH1 level was similarly measured via real-time polymerase chain reaction and Western blotting. The results showed that increased expression of miRNA-182, but neither miRNA-96 nor miRNA-183, could lead to higher expression levels in some hair cell markers. The morphology of the cells also did not change in this respect, but the evaluation of gene expression at the levels of mRNA could promote the expression of the ATOH1, SOX2, and POU4F3 markers. Furthermore, miRNA-182 could enhance the expression of ATOH1 at the protein level. According to the results of this study, it was concluded that miRNA-182 could serve as a crucial function in hair cell differentiation by the upregulation of SOX2, POU4F3, and ATOH1 to promote a hair cell’s fate.

scholarly journals The proportion of mitoses in different cell lineages changes during short-term culture of normal human bone marrow

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1240-1243
Author(s):  
M Keinanen ◽  
S Knuutila ◽  
CD Bloomfield ◽  
E Elonen ◽  
A de la Chapelle
Keyword(s):  
Bone Marrow ◽  
Cell Lineage ◽  
Great Majority ◽  
Hematopoietic Cell ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Immunofluorescent Staining ◽  
Cytoplasmic Staining ◽  
Cell Lineages ◽  
Mitotic Cells

To determine the hematopoietic cell lineage of mitotic cells in human bone marrow on direct examination and after 24-hour culture, marrow mitoses from four healthy individuals were studied, using a new technique that allows analysis of karyotypes in cells whose cell membrane and cytoplasm have been preserved. Mitoses were identified as being of erythroid lineage by immunofluorescent staining for surface glycophorin A and as being of granulocytic lineage by cytoplasmic staining for Sudan black B. On direct marrow examination without prior culture, the great majority of mitoses (74% to 90%) were of erythroid lineage; only a few (0% to 10%) were granulocytic. After 24-hour culture, the percentage of erythroid mitoses (15% to 40%) decreased, while the percentage of granulocytic mitoses (58% to 87%) increased strikingly. These data indicate that mitotic cells of different hematopoietic cell lineages predominate in marrow at different culture times and offer a plausible explanation for the high frequency of normal karyotypes in acute myeloid leukemia after direct marrow cytogenetic evaluation.

Macrophage inflammatory protein 1-alpha is a potential osteoclast stimulatory factor in multiple myeloma

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 671-675 ◽  
Author(s):  
Sun Jin Choi ◽  
Jose C. Cruz ◽  
Fiona Craig ◽  
Hoyeon Chung ◽  
Rowena D. Devlin ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Messenger Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Bone Marrow Cultures ◽  
Human Bone Marrow Cultures ◽  
Stimulatory Factor ◽  
Normal Controls ◽  
Marrow Cultures

Abstract This study was designed to determine if macrophage inhibitory protein-1 (MIP-1), a recently described osteoclast (OCL) stimulatory factor,1 was present in marrow from patients with multiple myeloma (MM) and possibly involved in the bone destructive process. MIP-1, but not interleukin-1β (IL-1β), tumor necrosis factor-β (TNF-β), or interleukin-6 (IL-6), messenger RNA was elevated in freshly isolated bone marrow from 3 of 4 patients with MM compared to normal controls. Furthermore, enzyme-linked immunosorbent assays of freshly isolated bone marrow plasma detected increased concentrations of hMIP-1 (range, 75-7784 pg/mL) in 8 of 13 patients (62%) with active myeloma, in 3 of 18 patients (17%) with stable myeloma (range, 75-190.3), as well as in conditioned media from 4 of 5 lymphoblastoid cell lines (LCLs) derived from patients with MM. Mildly elevated levels of MIP-1 were detected in 3 of 14 patients (21%) with other hematologic diagnoses (range, 80.2-118.3, median value of 96 pg/mL) but not in normal controls (0 of 7). MIP-1 was not detected in the peripheral blood of any patients with MM. In addition, recombinant hMIP-1 induced OCL formation in human bone marrow cultures. Importantly, addition of a neutralizing antibody to MIP-1 to human bone marrow cultures treated with freshly isolated marrow plasma from patients with MM blocked the increased OCL formation induced by these marrow samples but had no effect on control levels of OCL formation. Thus, high levels of MIP-1 are expressed in marrow samples from patients with MM, but not in marrow from patients with other hematologic disorders or controls, and support an important role for MIP-1 as one of the major factors responsible for the increased OCL stimulatory activity in patients with active MM.

scholarly journals Detection of residual lymphoma cells by polymerase chain reaction in peripheral blood is significantly less predictive for relapse than detection in bone marrow

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3800-3807 ◽  
Author(s):  
JG Gribben ◽  
D Neuberg ◽  
M Barber ◽  
J Moore ◽  
KW Pesek ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Peripheral Blood ◽  
Residual Disease ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Lymphoma Cells ◽  
Polymerase Chain ◽  
Residual Lymphoma ◽  
Minimal Residual

Abstract Polymerase chain reaction (PCR) amplification of the t(14;18) has been shown to be a highly sensitive method to detect minimal residual disease in patients with non-Hodgkin's lymphoma (NHL) whose tumors bear this translocation. The ideal tissue source to detect residual lymphoma would be from a previously involved lymph node. However, lymphoid tissue is rarely available once patients achieve complete remission. Although PCR amplification has been used to detect residual lymphoma cells in both bone marrow (BM) and peripheral blood (PB) of patients in complete remission, it is presently unknown whether BM and PB are equivalent tissue sources to detect residual disease. In the present study, we compared the clinical utility of the detection of residual lymphoma in both the BM and the PB of patients with advanced-stage non- Hodgkin's lymphoma before, at the time of, and after high-dose therapy and autologous BM transplantation (ABMT). The detection of residual lymphoma in either the BM or PB was associated with decreased disease- free survival. However, in the present study, 44% of patients who relapsed had no evidence of circulating lymphoma cells in their PB. At the time of BM harvest, PCR-detectable residual lymphoma cells were detected in 211 of 212 patients; although, in a subset of these patients analyzed, lymphoma cells were detected in the peripheral blood of only 49% of patients. When residual lymphoma cells within the autologous BM are infused into the patient these cells are rapidly detectable circulating in the PB in the patient. These cells continue to circulate during the immediate posttransplant period and be detectable in the PB in the majority of patients who are infused with marrow containing residual lymphoma. We conclude that BM is a more informative tissue source than PB in detecting minimal residual disease at the time of and after ABMT, and that contamination of PB early after ABMT appears to be the consequence of reinfusion of lymphoma cells within autologous marrow.

New Somatic Mutation in the PIG-A Gene Emerges at Relapse of Paroxysmal Nocturnal Hemoglobinuria

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3422-3427 ◽  
Author(s):  
Khédoudja Nafa ◽  
Monica Bessler ◽  
H. Joachim Deeg ◽  
Lucio Luzzatto
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Longitudinal Study ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Single Base ◽  
Single Base Pair ◽  
Polymerase Chain ◽  
American Society

We report a detailed longitudinal study of the first patient to be treated (in 1973) for paroxysmal nocturnal hemoglobinuria (PNH) with syngeneic bone marrow transplantation (BMT). The patient subsequently relapsed with PNH in 1983, and still has PNH to date. Analysis of thePIG-A gene in a recent blood sample showed in exon 6 an insertion-duplication causing a frameshift. Polymerase chain reaction (PCR) amplification of the PIG-A exon 6 from bone marrow (BM) slides obtained before BMT showed that the duplication was not present; instead, we found several single base pair substitutions in exons 2 and 6. Thus, relapse of PNH in this patient was not due to persistence of the original clones; rather, it was associated with the emergence of a new clone. These findings support the notion that the BM environment may create selective conditions favoring the expansion of PNH clones.© 1998 by The American Society of Hematology.

The role of platelet α-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3094-3101 ◽  
Author(s):  
Ranita Sungaran ◽  
Orin T. Chisholm ◽  
Boban Markovic ◽  
Levon M. Khachigian ◽  
Yoshihiro Tanaka ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Mrna Expression ◽  
Stromal Cells ◽  
Messenger Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Dependent Manner ◽  
Platelet Factor ◽  
Dose Dependent

Thrombopoietin (TPO), the specific cytokine that regulates platelet production, is expressed in human bone marrow (BM), kidney, and liver. There appears to be no regulation of TPO in the kidney and liver, but TPO messenger RNA (mRNA) expression can be modulated in the stromal cells of the BM. In this study, we used primary human BM stromal cells as a model to study the regulation of TPO mRNA expression in response to various platelet -granular proteins. We showed that platelet-derived growth factor (PDGF) BB and fibroblast growth factor (FGF) 2 stimulated TPO mRNA expression in both a dose-dependent and time-dependent manner. The addition of 50 ng/mL of PDGF and 20 ng/mL of FGF resulted in maximal induction of TPO mRNA expression in 4 hours. We also found that platelet factor 4 (PF4), thrombospondin (TSP), and transforming growth factor-beta (TGF-β) are negative modulators of megakaryocytopoiesis. We observed suppression in TPO mRNA expression with 1 μg/mL of both PF4 and TSP and 50 ng/mL of TGF-β, with maximal suppression occurring 4 hours after the addition of these proteins. Finally, the addition of whole-platelet lysate produced a dose-dependent inhibition of TPO expression. On the basis of these findings, we propose that the platelet -granular proteins studied may regulate TPO gene expression in BM stromal cells by means of a feedback mechanism.

Macrophage inflammatory protein 1-alpha is a potential osteoclast stimulatory factor in multiple myeloma

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 671-675 ◽  
Author(s):  
Sun Jin Choi ◽  
Jose C. Cruz ◽  
Fiona Craig ◽  
Hoyeon Chung ◽  
Rowena D. Devlin ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Messenger Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Bone Marrow Cultures ◽  
Human Bone Marrow Cultures ◽  
Stimulatory Factor ◽  
Normal Controls ◽  
Marrow Cultures

This study was designed to determine if macrophage inhibitory protein-1 (MIP-1), a recently described osteoclast (OCL) stimulatory factor,1 was present in marrow from patients with multiple myeloma (MM) and possibly involved in the bone destructive process. MIP-1, but not interleukin-1β (IL-1β), tumor necrosis factor-β (TNF-β), or interleukin-6 (IL-6), messenger RNA was elevated in freshly isolated bone marrow from 3 of 4 patients with MM compared to normal controls. Furthermore, enzyme-linked immunosorbent assays of freshly isolated bone marrow plasma detected increased concentrations of hMIP-1 (range, 75-7784 pg/mL) in 8 of 13 patients (62%) with active myeloma, in 3 of 18 patients (17%) with stable myeloma (range, 75-190.3), as well as in conditioned media from 4 of 5 lymphoblastoid cell lines (LCLs) derived from patients with MM. Mildly elevated levels of MIP-1 were detected in 3 of 14 patients (21%) with other hematologic diagnoses (range, 80.2-118.3, median value of 96 pg/mL) but not in normal controls (0 of 7). MIP-1 was not detected in the peripheral blood of any patients with MM. In addition, recombinant hMIP-1 induced OCL formation in human bone marrow cultures. Importantly, addition of a neutralizing antibody to MIP-1 to human bone marrow cultures treated with freshly isolated marrow plasma from patients with MM blocked the increased OCL formation induced by these marrow samples but had no effect on control levels of OCL formation. Thus, high levels of MIP-1 are expressed in marrow samples from patients with MM, but not in marrow from patients with other hematologic disorders or controls, and support an important role for MIP-1 as one of the major factors responsible for the increased OCL stimulatory activity in patients with active MM.

scholarly journals Isolation of an early T-cell precursor (CFU-TL) from human bone marrow

Blood ◽  
1990 ◽  
Vol 75 (5) ◽  
pp. 1064-1068 ◽  
Author(s):  
JM Bertho ◽  
MD Mossalayi ◽  
AH Dalloul ◽  
G Mouterde ◽  
P Debre
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
High Capacity ◽  
Cell Lineage ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Differentiation Potential ◽  
T Cell Clones ◽  
Cell Clones

Abstract CD2-CD3-CD4-CD8- human bone marrow (BM) cells were previously shown to generate T-cell clones in vitro. This capacity was abolished after treatment of this population with anti-CD7 monoclonal antibody and complement. In this study, using rosetting with sheep erythrocytes, complement-dependent cytotoxicity, and specific immunoadherence method, we isolated a minor BM subset that contained more than 80% CD7+CD2-CD3- CD4-CD8- cells with small lymphoid cell morphology. They comprised most early T-cell precursors (CFU-TL) as they displayed high capacity to generate T-cell clones when cultured in limiting dilutions. CFU-TL nature of these cells was also confirmed by the sequential expression of mature T-cell specific markers on their surface after in vitro induction. This BM subset also contained 2% to 3% CFU-GM precursors. Together, these results pointed to the existence of BM CD7+CD2- precursors with high differentiation potential and showed the commitment of most of them to T-cell lineage.

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