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PubMed1977

Constant post-irradiation repopulation rates and linear relationship between cellular blood response and number of transplanted bone marrow cells in inbred mice.

Graded doses of syngeneic bone marrow cells were transplanted into lethally irradiated mice. Repopulation curves of peripheral blood granulocytes and platelets were apparently exponential and parallel after doses larger than 5 X 10(5) cells. The blood platelet Td was reduced from 111 h to 53-57 h, and granulocyte Td from 57 to 40 h in transplanted groups. The mean blood cell counts were reproducible enough to be used as a biological assay of the amount of bone marrow cells transplanted. Linear relationship between increment of blood cells up to day 16 and number of bone marrow cells transplanted on day 1 was demonstrated (1,200 granulocytes and 14,300 platelets/mul blood per 10(5) bone marrow cells). The linearity suggested a mean Td less than 22.5 h of proliferating bone marrow cells, and allowed a rough estimation of mouse bone marrow stem cell radiosensitivity (Do 76 rad).

Animals↗
PubMed1987

Differential repair of gamma-ray-induced DNA strand breaks by various cellular subpopulations of mouse jejunal epithelium and bone marrow in vivo.

We have examined the induction and repair of gamma-ray-induced DNA strand breaks in different subpopulations of cells in mouse jejunal epithelium and bone marrow using a modification of the alkaline elution methodology whereby different populations of cells are selectively labeled with radioactive DNA precursors. Mice were labeled by intraperitoneal injection with between 0.5 and 2.0 mu Ci/g of [3H]thymidine at various times prior to irradiation with 10 Gy of gamma rays. In the studies with jejunal epithelium, the timing of the injection of the radiolabel relative to the irradiation was varied between 6 and 72 h, depending on the cell population of interest. The DNA damage and repair characteristics representative of both the total cell population and the radiolabeled fraction of these cells were then measured. Little difference was noted in the amount of initial damage induced in these different populations of cells. However, for both the jejunum and bone marrow, cells that incorporated the radiolabel within 6 h after injection (i.e., rapidly proliferating cells) repaired their strand breaks more rapidly than did the remainder of the population. In the case of jejunum, the repair capacity of the radiolabeled cell population progressively diminished as the cells matured and differentiated so that cells that contained the radiolabel 72 h after injection (i.e., mature villus cells) actually repaired their strand breaks more slowly than did the bulk cells.

Animals↗
PubMed1985

Diagnostic value of bone marrow imaging with 111indium-transferrin and 99m technetium-colloids in myelofibrosis.

111Indium--transferrin (111In) and 99mTechnetium-colloids (99mTc) bone marrow imaging of 55 myelofibrosis (MF) cases has been compared with clinical, histological, and iron-kinetics data. The best correlations are seen between the splenic uptake of 111In with the spleen/sacrum ratio of 59Fe at the first hour (r = 0.69, P less than 0.001) and also with the splenic erythropoiesis histologically assessed in ten splenectomized patients (r = 0.75, P less than 0.01). Moreover, sacrum uptake of 111In, when compared with sacrum uptake of 59Fe (r = 0.51, P less than 0.001) and with hematopoietic cellularity of the bone marrow (r = 0.57, P less than 0.001) reflects faithfully the hematopoietic cell content of the marrow. Thus, 111In bone marrow imaging provides a noninvasive and useful tool for the diagnosis of myeloid metaplasia in MF. Ferrokinetic studies still appreciate with better insight the amount of ineffective erythropoiesis or hemolysis and remains therefore of great value when splenectomy is discussed.

Adult↗
PubMed1988

Simultaneous analysis of DNA content and surface antigens in human bone marrow.

In order to identify when cellular expansion occurs during hematopoietic maturation, a method was developed for the simultaneous analysis of one or two cell-surface antigens and DNA content on bone marrow cells while preserving their light-scatter properties. Proliferation in a population defined by light-scatter and surface-antigenic characteristics was assessed by measuring the percentage of cells in this population having more than 2C amount of DNA ("proliferation index"). Viable, low-density (1.077 g/cm3), bone marrow cells, stained with monoclonal antibodies conjugated with fluorescein or phycoerythrin, were fixed with paraformaldehyde and subsequently treated with the detergent, Tween 20. The UV-excitable DNA stain Hoechst 33342 was used to quantify DNA content in the cells without interference with immunofluorescence. A FACS IV flow cytometer was used, equipped with the first laser at 488 nm emitting for light scattering and immunofluorescence measurements and the second laser emitting at 360 nm for the Hoechst excitation. The Hoechst uptake was the same for all bone marrow populations, yielding a tight coefficient of variation (CV) (average 5.0%) for the G0/G1 DNA peak. This permitted high sensitivity of cell detection in S, G2, and M phases of the cell cycle, while preserving light-scattering properties of the cells and maintaining cell surface immunofluorescence. The lowest "proliferation index" detected using this technique was 0.08% in a sample obtained from a patient with chronic lymphocytic leukemia. Normal helper T lymphocytes in marrow had approximately 0.5% of the cells in S, G2, or M phase. We show that the erythroid lineage, in the adult normal bone marrow, is the most active in proliferation among all hematopoietic lineages.

Adult↗
PubMed2005

Matrix metalloproteinase-9 modulation by resident arterial cells is responsible for injury-induced accelerated atherosclerotic plaque development in apolipoprotein E-deficient mice.

OBJECTIVE: Although matrix metalloproteinase-9 (MMP-9) has been implicated in atherosclerotic plaque instability, the exact role it plays in the plaque development and progression remains largely unknown. We generated apolipoprotein E (apoE)-deficient (apoE-/-) MMP-9-deficient (MMP-9-/-) mice to determine the mechanisms and the main cell source of MMP-9 responsible for the plaque composition during accelerated atherosclerotic plaque formation. METHODS AND RESULTS: Three weeks after temporary carotid artery ligation revealed that while on a Western-type diet, apoE-/- MMP-9-/- mice had a significant reduction in intimal plaque length and volume compared with apoE-/- MMP-9+/+ mice. The reduction in plaque volume correlated with a significantly lower number of intraplaque cells of resident cells and bone marrow-derived cells. To determine the cellular origin of MMP-9 in plaque development, bone marrow transplantation after total-body irradiation was performed with apoE-/- MMP-9+/+ and apoE-/- MMP-9-/- mice, which showed that only MMP-9 derived from resident arterial cells is required for plaque development. CONCLUSIONS: MMP-9 is derived from resident arterial cells and is required for early atherosclerotic plaque development and cellular accumulation in apoE-/- mice.

Animals↗
PubMed1988

Bone marrow biopsy during induction chemotherapy for acute myeloid leukaemia identifies only 50% of patients with resistant disease.

A study was carried out to determine whether bone marrow biopsy performed on day 6 of induction therapy for acute myeloid leukaemia (AML) can identify those patients with resistant disease who would need an intensification of the first course of induction. Bone marrow biopsies were performed on day 6 of induction chemotherapy in 44 patients with AML treated with daunorubicin, cytosine arabinoside and thioguanine. Biopsies were assessed for blast count, trephine cellularity and leukaemic index. Discrimination between patients who went on to achieve remission and those with resistant disease was best achieved using the reduction in bone marrow cellularity from pretreatment marrow to day-6 marrow. However, this discriminator identified only 50% of the patients with resistant disease and included 13% of patients who achieved remission with the first course of chemotherapy. The other parameters of response were even less effective at discriminating between chemotherapy-resistant and chemotherapy-responsive disease.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2001

Incidence of fat embolism syndrome after cemented or cementless bilateral simultaneous and unilateral total knee arthroplasty.

This prospective study enrolled 100 patients undergoing unilateral total knee arthroplasty (TKA) and 100 patients undergoing bilateral simultaneous TKA. Bilateral simultaneous TKAs were done under 1 anesthesia with one side immediately following the other. To determine hemodynamic changes and to detect fat and bone marrow embolization, arterial and right atrial blood samples were obtained before insertion (baseline) and 1, 3, 5, and 10 minutes after insertion of the femoral alignment rod. Arterial and right atrial blood samples were obtained 1, 3, 5, and 10 minutes after insertion of the tibial component broach. Blood samples were obtained at 24 and 48 hours after the operation. Arterial blood pressure, heart rate, right atrial pressure, arterial oxygen tension, and carbon dioxide tension were monitored at corresponding times. The presence of fat was determined with oil red O fat stain, and the presence of cellular contents of bone marrow was determined with Wright-Giemsa stain. Fat embolism was found in 65 patients (65%) with a bilateral TKA and 46 patients (46%) with a unilateral TKA. Bone marrow cell embolism was found in 12 patients (12%) with a bilateral TKA and in 4 patients (4%) with a unilateral TKA. Six patients with positive bone marrow cells (2 patients with a unilateral TKA and 4 patients with a bilateral TKA) had neurologic manifestations.

Adult↗
PubMed1990

A 17-kd polypeptide, sensitive to bacterial collagenase, is synthesized by bone marrow macrophages.

A novel polypeptide with a molecular weight of 17 kd (17k protein) was identified in bone marrow cell cultures. The synthesis of 17k protein is elevated in cell cultures maintained under Dexter conditions, which support myelopoiesis. The predominance of macrophages in the stromal layer of these cultures and the observation that a mouse myelomonocytic cell line P388D1 is capable of synthesizing large amounts of 17k protein led us to the study of its synthesis by bone marrow macrophages. Metabolic labeling with [14C]proline and partial amino acid analysis of 17k protein demonstrated that the polypeptide contains relatively high amounts of proline and is also sensitive to degradation with bacterial collagenase. However, no hydroxyproline is detectable in 17k protein, and it is extensively degraded with bacterial collagenase. However, no hydroxyproline is detectable in 17k protein, and it is extensively degraded by proteolysis with pepsin, using conditions under which collagen triple helices are resistant to degradation, suggesting that collagen-like structures are not contained in 17k protein. This polypeptide is found predominantly in the cellular layers of bone marrow macrophage cultures. Incorporation of [14C]proline into 17k protein is diminished by increasing concentrations of colony-stimulating factor 1 (CSF-1). The 17k protein may be involved in macrophage proliferation because its synthesis is inhibited by CSF-1, which is required for the maintenance of bone marrow macrophages in vitro.

Animals↗
PubMed2000

[New reticulocyte indices and their utility in hematologic diagnosis].

Reticulocyte count in manual method has been the assay traditionally used to evaluate the status of erythropoiesis in haematological disorders with disturbances in erythropoietic activity. But it is a qualitative rather than a quantitative test due to the high variability in manual microscopic method. Automated reticulocyte counting based on flow cytometry has provided much objective and exactly measure of percentage and absolute number of reticulocytes than microscopic method. Besides these two traditional parameters automatic reticulocyte counters can detect differences in the amounts of cellular RNA present in red blood cells which reflects their maturational stages. There are three related parameters which describes the reticulocyte maturation: RMI (reticulocyte maturity index), HFR (high fluorescence reticulocytes), IRF (immature reticulocyte fraction). These parameters can be used as the earliest signs of marrow engraftment after autologous or allogeneic bone marrow transplantations. Reticulocyte cellular indices such as MCVr (mean reticulocyte volume), CHr (reticulocyte hemoglobin content), CHCMr (reticulocyte hemoglobin concentration) entered recently in few automated reticulocyte counters allows for assessment of the functional state of the erythropoiesis in the diagnosis and monitoring of the iron deficiency and rhEpo therapy. Although there is no agreement between various methods of staining and counting of the reticulocytes and lack of standardization materials for this assay automated reticulocyte counting and new reticulocyte parameters probably will improve the diagnosis and monitoring of many haematological diseases.

Erythropoiesis↗
PubMed2005

Response criteria for myelofibrosis with myeloid metaplasia: results of an initiative of the European Myelofibrosis Network (EUMNET).

The European Myelofibrosis Network (EUMNET), a European research network on myelofibrosis with myeloid metaplasia (MMM), has developed a definition of response for the disease by using clinicohematologic, histologic, and cytogenetic criteria. A core set of 5 clinicohematologic criteria was selected out of 9 candidates on the basis of their sensitivity to change measured in 196 patients treated either during clinical trials or routine clinical practice. A consensus panel of 16 international experts was convened and asked to score the level of response in 104 patient profiles as major, moderate, minor, or no response according to changes of the clinicohematologic criteria. Using the experts' consensus as the gold standard, the performance of 100 possible definitions of response was evaluated. Criteria for major or moderate clinicohematologic response were determined to be changes in hemoglobin (Hb) and spleen size and the presence of constitutional symptoms, while changes in platelet count and white blood cell (WBC) count served as complementary criteria and were of value for defining minor responses. A histologic response was defined by changes in bone marrow fibrosis and cellularity grades. The combined use of these response definitions should help standardize the design and reporting of future clinical studies in MMM.

Adult↗
PubMed1999

Different effect of granulocyte colony-stimulating factor or bacterial infection on bone-marrow cells of cyclophosphamide-treated or irradiated mice.

In the present study, the effect of treatment with granulocyte colony-stimulating factor (G-CSF) on cellular composition of the bone marrow and the number of circulating leucocytes of granulocytopenic mice, whether or not infected with Staphylococcus aureus, was assessed. With two monoclonal antibodies, six morphologically distinct cell populations in the bone marrow could be characterised and quantitated by two-dimensional flow cytometry. Granulocytopenia was induced by cyclophosphamide or sublethal irradiation. Cyclophosphamide predominantly affected the later stages of dividing cells in the bone marrow resulting in a decrease in number of granulocytic cells, monocytic cells, lymphoid cells and myeloid blasts. G-CSF administration to cyclophosphamide-treated mice increased the number of early blasts, myeloid blasts and granulocytic cells in the bone marrow, which indicates that this growth factor stimulates the proliferation of these cells in the bone marrow. During infection in cyclophosphamide-treated mice the number of myeloid blasts increased. However, when an infection was induced in cyclophosphamide and G-CSF-treated mice, the proliferation of bone-marrow cells was not changed compared to that in noninfected similarly treated mice. Sublethal irradiation affected all bone-marrow cell populations, including the early blasts. G-CSF-treatment of irradiated mice increased only the number of myeloid blasts slightly, whereas an infection in irradiated mice, whether or not treated with G-CSF, did not affect the number of bone-marrow cells. Together, these studies demonstrated that irradiation affects the early blasts and myeloid blasts in the bone marrow more severely than treatment with cyclophosphamide. Irradiation probably depletes the bone marrow from G-CSF-responsive cells, while cyclophosphamide spared G-CSF responsive cells, thus enabling the enhanced G-CSF-mediated recovery after cyclophosphamide treatment. Only in these mice, bone marrow recovery is followed by a strong mobilisation of mature granulocytes and their band forms from the bone marrow into the circulation during a bacterial infection.

Agranulocytosis↗
PubMed1985

Xenogeneic monoclonal antibody to an Ly-6-linked murine cell surface antigen: differential reactivity with T cell subpopulations and bone marrow cells.

The patterns of cellular and strain reactivity of a monoclonal antibody (6C3 MAb) derived from the fusion of SP2/0 cells with splenocytes from rats immunized against MRL/MpJ-lpr/lpr T cells were characterized by using flow cytofluorometry (FCF) analysis. This MAb was found to stain 70 to 90% of T cells of mice with the lpr/lpr genotype and 20 to 60% of T cells of congenic +/+ strains. Dual-parameter FCF analysis of Lyt-2 vs 6C3 expression revealed the existence of several Lyt-2- and Lyt-2+ T cell subsets, one of which (Lyt-2- bright 6C3+) was expanded in lpr/lpr-bearing mice. The 6C3 MAb stained only 2 to 5% normal thymocytes but reacted with 40 to 50% bone marrow (BM) cells. A strain survey demonstrated the expression of the 6C3 antigen on peripheral T cells (and BM cells) of all strains examined, with the exception of NOD, NZB/B1NJ, and ST/bJ. Interestingly, in the positive strains, two types of 6C3 staining patterns of T cells were observed: bimodal or trimodal. Study of BXH and CXB recombinant inbred (RI) strains demonstrated that the bimodal and trimodal 6C3 patterns are associated with the Ly-6.1 and Ly-6.2 phenotypes, respectively. Linkage of 6C3 expression with the Ly-6 locus was confirmed by using the congenic C3H.B6-Ly-6b strain. Moreover, the 6C3 staining of T cells in Ly-6.2 strains was reduced by preincubation with the H9/25 and SK-142-446 MAb, which are known to recognize Ly-6.2-associated antigens. Therefore, the 6C3 MAb appears to detect a frame-work determinant on an Ly-6-linked antigen that is absent from T cells of NOD, NZB, and ST/bJ mice. Analysis of (NZB x C58) NX8 RI strains demonstrated a correlation between the lack of 6C3 expression on T cells and unresponsiveness in autologous mixed lymphocyte reaction (a property of NZB/B1NJ mice). The 6C3 MAb should prove useful for further genetic and biochemical analysis of the Ly-6 locus and its product(s), and for the delineation of functional subsets of T cells and BM cells in normal and lpr/lpr-bearing mice.

Animals↗