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PubMed2006

Histological and cytogenetic characterization of bone marrow in relation to prognosis and diagnosis of myelodysplastic syndromes.

Bone marrow (BM) histology of 102 myelodysplastic syndromes (MDS) patients was analyzed retrospectively. All the cases were reclassified according to the World Health Organization (WHO) classification. Karyotype study was conducted for all except one. Fifteen of the MDS cases were hypoplastic. The cellularity in bone marrow histology is sometimes ineffective in the differential diagnosis of MDS and aplastic anemia (AA). Nonetheless, a marked decrease in the number of megakaryocytes (average, 0.3/mm(2); range, 0-2/mm(2)) even in the hyperplastic foci of the marrow of AA was the most important histological feature differentiating AA from MDS, whereas the number of megakaryocytes increased in most MDS cases (44/mm(2); range, 1-240/mm(2)) and also in hypoplastic MDS (14/mm(2); range, 8-26/mm(2)). Hyperplastic marrow had a significantly high frequency of progress to acute myeloid leukemia (AML) and hypoplastic MDS had a lower rate of progress to AML. Severe myelofibrosis had a significantly poor prognosis. An increase in CD34-positive cells in MDS indicated a high rate of progress to AML. As for the patients with refractory cytopenia with multilineage dysplasia (RCMD; the new category under the WHO classification), the increased number of megakaryocytes was correlated with poor prognosis.

Adult↗
PubMed2004

Ehrlich ascites tumor as a tool in the development of compounds with immunomodulatory properties.

In previous works, we have demonstrated that the myeloprotective properties of several natural and synthetic compounds are partly responsible for their antitumor activity in the Ehrlich ascites tumor (EAT) model. In this work, we present information that may be useful to the study of pharmacological and toxicological properties of compounds that affect the hematological compartment. Clonogenic studies in EAT-inoculated mice demonstrated a rapid decrease in bone marrow CFU-GM, whereas a progressive increase in splenic CFU-GM and cellularity was observed, followed by splenomegaly. Bone marrow cellularity declined on the third day after tumor challenge, returning to normal values thereafter. Serum from EAT-bearing mice produced detectable colony-stimulating activity in vitro. Similar results were observed with the conditioned medium from Ehrlich tumor cell cultures, but not with the cell-free Ehrlich tumor ascitic fluid. Tumor inoculation also resulted in a more striking depletion in the number of non-adherent cells in long-term bone marrow cell cultures (LTBMCs) with no bone marrow stroma formation. We speculate that the physiological alterations induced by the EAT growth can be used to assess the ability of compounds to modulate the hematopoietic response.

Animals↗
PubMed1981

[Acute neutropenia in adult severe bacterial infections. Six cases (author's transl)].

A transient neutropenia (duration : less than 48 hours) occurred in 6 patients with severe bacterial infections. The bone marrow myeloïd cellularity was high or normal; a maturation arrest of the myeloïd cells - at a variable stage - was present in each case, as well as circulating granulocyte precursors. Other causes of neutropenia were excluded. A general survey of the literature is made and the mechanisms of this bacterial neutropenia are considered : bone marrow failure seems mostly improbable ; the few previously reported cases, the experimental bacterial neutropenias, the peculiar, transient, evolution we observed, all favor a peripheral mechanism ; the bacterial infection seems to act by; two ways : in septicemia (especially during septic shock) by enhancing leucocyte margination along vascular walls ; in large septic foci (mostly pulmonary) by an important destruction of polymorphonuclear neutrophils. Such bacterial infections share a bad prognosis but the neutropenia itself needs no specific treatment.

Acute Disease↗
PubMed1997

Aspiration to obtain osteoblast progenitor cells from human bone marrow: the influence of aspiration volume.

UNLABELLED: Bone marrow contains osteoblast progenitor cells that can be obtained with aspiration and appear to arise from a population of pluripotential connective-tissue stem cells. When cultured in vitro under conditions that promote an osteoblastic phenotype, osteoblast progenitor cells proliferate to form colonies of cells that express alkaline phosphatase and, subsequently, a mature osteoblastic phenotype. We evaluated the number of nucleated cells in bone-marrow samples obtained with aspiration from the anterior iliac crest of thirty-two patients without systemic disease. There were nineteen male patients and thirteen female patients; the mean age was forty-one years (range, fourteen to seventy-seven years). The prevalence and concentration of the osteoblast progenitor cells also were determined, by placing the bone-marrow-derived cells into tissue-culture medium and counting the number of alkaline phosphatase-positive colony-forming units. In order to assess the effect of aspiration volume, two sequential experiments were performed. In the first experiment, aspiration volumes of one and two milliliters were compared. In the second experiment, aspiration volumes of two and four milliliters were compared. The mean prevalence of alkaline phosphatase-positive colony-forming units in the bone-marrow samples was thirty-six per one million nucleated cells (95 per cent confidence interval, 28 to 47); a mean of 2400 alkaline phosphatase-positive colony-forming units was obtained from a two-milliliter aspirate. There was a significant difference among the patients with respect to the number of alkaline phosphatase-positive colony-forming units in these bone-marrow samples (p < 0.001). Seventy per cent of this variation in the prevalence was due to variation among patients, and 20 per cent was due to variation among aspirates. The number of alkaline phosphatase-positive colony-forming units in the aspirate increased as the aspiration volume increased. However, contamination by peripheral blood also increased as the aspiration volume increased. An increase in the aspiration volume from one to four milliliters caused a decrease of approximately 50 per cent in the final concentration of alkaline phosphatase-positive colony-forming units in an average sample. CLINICAL RELEVANCE: On the basis of these data, we recommend that, when bone marrow is obtained with aspiration for use as a bone graft, the volume of aspiration from any one site should not be greater than two milliliters. A larger volume decreases the concentration of osteoblast progenitor cells because of dilution of the bone-marrow sample with peripheral blood. We estimate that four one-milliliter aspirates will provide almost twice the number of alkaline phosphatase-positive colony-forming units as will one four-milliliter aspirate. In addition, these data confirm that humans differ significantly from one another with respect to the cellularity of bone marrow and the prevalence of osteoblast progenitor cells. Additional studies are necessary to determine if the number or prevalence of alkaline phosphatase-positive colony-forming units in bone marrow is a determining factor in the efficacy of an autogenous bone or bone-marrow graft and to ascertain how the number and function of alkaline phosphatase-positive colony-forming units may change as a function of factors such as age, menopausal status, and selected diseases.

Adolescent↗
PubMed1992

Morphology in patients with severe aplastic anemia treated with antilymphocyte globulin.

One hundred and seventeen patients with severe aplastic anemia (SAA) were treated at our institution between 1976 and 1990 with antilymphocyte globulin (ALG) therapy. Seventy-nine (68%) are alive and probability of survival at 14 years, according to Kaplan and Meier, is 62% +/- 12%. Twenty-six patients developed a late clonal complication: 11 had a myelodysplastic syndrome (MDS) and 17 had paroxysmal nocturnal hemoglobinuria (PNH); two patients had both. The cumulative risk at 10 years is 42%. The development of MDS/PNH after SAA directly affects survival. The probability of being alive at 14 years is 81% +/- 10% for patients with stable disease and 36% +/- 13% for those with clonal evolution (P = .001). To look for predictive signs, we reevaluated peripheral blood and bone marrow cytomorphology at presentation, during regeneration, and in remission. We examined the peripheral blood values for hemoglobin, reticulocytes, granulocytes, thrombocytes, mean corpuscular volume (MCV), and fetal hemoglobin, as well as bone marrow for cellularity, erythropoiesis, myelopoiesis, and megakaryopoiesis. ALG therapy induces slow and incomplete recovery. Although in "remission," ALG patients have lower hemoglobin values, higher reticulocyte counts, lower granulocyte and platelet values, and a higher MCV and fetal hemoglobin than normal controls. They retain a reduced number of megakaryocytes and a persistence of atypical monocytes in bone marrow morphology as stigmata of their disease. Patients with late clonal complications show distinct morphologic abnormalities: patients with PNH have higher MCVs, higher granulocyte and reticulocyte counts, and more dyserythropoiesis at diagnosis and a lower hemoglobin with an increased proportion of erythroblasts in the bone marrow in "remission." Patients who later developed MDS are not different from the total patient population at diagnosis. After therapy, these patients are characterized by the presence of ring sideroblasts and atypical monocytes during regeneration and by a persistent increase in MCV, a higher fetal hemoglobin, lower granulocyte values, and megakaryocytic dysplasia during "remission." Thus, routine morphologic follow-up examination of blood and bone marrow can discover patients at risk for late hematologic complications after ALG therapy.

Adult↗
PubMed2004

Distinct phases in recovery of reconstituted innate cellular-mediated immunity after murine syngeneic bone marrow transplantation.

Defects in immune reconstitution after hematopoietic stem cell transplantation confer extreme infection risk on to the transplant recipient. Perturbations in adaptive immune reconstitution have been well characterized, yet defects in reconstituted innate cellular-mediated immunity remain largely unstudied. Recovery in innate effector cells was defined by using an established murine model of autologous bone marrow transplantation. Cytokine induction after cell culture and systemic stimulation with pathogen-associated molecular patterns was also measured for control, transplant-recipient, and irradiated-only animals. Early reconstitution (7 to 14 days) of donor-derived macrophages, dendritic cells, and polymorphonuclear cells was associated with recovery in interleukin (IL)-12p70 and IL-6 production. Later reconstitution (21 days) of natural killer cells was associated with interferon (IFN)-gamma recovery. Hence, splenocyte innate cellular-mediated immunity recovered to normal levels in cellularity and IL-12p70, IFN-gamma, and IFN-alpha production by 21 days after transplantation. In contrast, levels of systemic cytokine production from transplant-recipient and irradiated-only animals were preserved despite incomplete or absent hematopoietic reconstitution. These results suggest that innate immune responses to systemic inflammatory challenges are largely intact after autologous bone marrow transplantation, whereas local innate cellular-mediated immunity within reconstituting lymphoid organs may be impaired. The disparate effects of autologous hematopoietic stem cell transplantation on host immune function may translate to differences in susceptibility to local versus systemic infectious challenges.

Animals↗
PubMed1974

The cellular composition of the granulocyte series in the normal human bone marrow according to the volume of the sample.

The cellular composition of the granulocytic series in normal human bone marrow was studied in aspirates of different volume. An increase in the proportion of polymorphonuclears was observed in samples of more than 1 ml, thus indicating a dilution of bone marrow by peripheral blood. Certain data from the literature, referring to the differential count in the bone marrow granulocytic series, are discussed. The differences between these data can be explained by the dilution of bone marrow cells by mature granulocytes in blood.

Biopsy, Needle↗
PubMed2005

Criteria for defining a complete remission in acute myeloid leukaemia revisited. An analysis of patients treated in HOVON-SAKK co-operative group studies.

Complete remission (CR) in patients with acute myeloid leukaemia (AML) is the primary endpoint for the evaluation of induction treatment and treatment strategies. However, the choice and application of the criteria for a haematological CR can often become a subject of debate because of regeneration more than 5% blasts may be present at the time of response evaluation; platelet and neutrophil recovery may be incomplete and marrow cellularity can vary. This study examined the individual parameters for CR in 1250 adult patients with de novo AML treated according to three successive study protocols. Patients with < or =5% blasts showed the best overall survival (OS) and the lowest relapse risk (RR). This was independent of blast cells present in the peripheral blood or bone marrow (BM) cellularity. In the same patient group, the presence of extramedullary leukaemia, incomplete platelet (<100 x 10(9)/l) or neutrophil (<1.0 x 10(9)/l) recovery caused a reduced OS and increased RR. In conclusion, < or =5% blasts in the BM, recovery of neutrophils and platelets, and the absence of extramedullary disease constitute the cornerstones for the definition of a haematological CR in patients with AML.

Acute Disease↗
PubMed2007

Osteoporosis of hematologic etiology.

OBJECTIVE: Here we present evidence that overexertion of the hematopoietic system following chronic bleeding plays an important role in the etiology of osteoporosis. MATERIALS AND METHODS: C57BL/6 mice were exposed to chronic bloodletting (0.2 mL twice per month for 10 months), total body irradiation (900 cGy), or aging (20-30 months old). Bone marrow from standard untreated donors was transplanted under the kidney capsules of all three categories of recipients to investigate the influence of each of these conditions on new bone marrow formation. Cellularity and histologic structure of developed osteohematopoietic sites and histomorphometry of lumbar vertebrae were studied, thus assessing the role of bleeding, irradiation, and old age on new bone formation and effects on existing bone. RESULTS: Chronic blood loss led to augmented production of hematopoietic microenvironment, relative reduction in the amount of generated bone, and activation of the bone resorptive process in the newly forming osteohematopoietic complex. Similar results were seen in irradiated and senescent mice. Activity, stimulating expansion of hematopoietic microenvironment, was revealed in the plasma of all three categories of experimental mice. Likewise, quantification of the relative amount of bone and hematopoietic areas in skeletal sites showed a significant reduction in bone tissue of the first lumbar vertebrae of chronically bled mice. CONCLUSIONS: Our experimental data, together with existing clinical observations documenting the role of hematopoietic insufficiency in the development of osteoporosis, confirm our working hypothesis that chronic blood loss may be the primary factor responsible for the rapid and consistent development of postmenopausal osteoporosis.

Age Factors↗
PubMed2004

Circadian regulation of cell cycle and apoptosis proteins in mouse bone marrow and tumor.

Proapoptotic drugs such as docetaxel displayed least toxicity and highest antitumor efficacy following dosing during the circadian rest phase in mice, suggesting that cell cycle and apoptotic processes could be regulated by the circadian clock. In study 1, mouse bone marrow and/or tumor were obtained every 4 h for 24 h in C3H/HeN mice with or without MA13/C mammary adenocarcinoma in order to determine the circadian patterns in cell-cycle phase distribution and BCL-2 anti-apoptotic protein expression. In study 2, mouse bone marrow from B6D2F1 mice was sampled every 3 h for 24 h in order to confirm the BCL-2 rhythm and to study its relation with 24 h changes in the expression of proapoptotic BCL-2-associated X protein (BAX) protein and clock genes mPer2, mBmal1, mClock, and mTim mRNAs. The rhythms in G1-, S- or G2/M-phase cells were shifted in tumor compared with bone marrow. In the tumor, the mean proportion of G2/M-phase cells increased by 75% from late rest to late activity span (P from cosinor = 0.001). No 24 h rhythm was found for BCL-2 in tumors. In contrast to this, in the bone marrow, mean BCL-2 expression varied 2.8-fold in B6D2F1 mice (P=0.025) and 3- or 4.5-fold in tumor-bearing and nontumor-bearing C3H/HeN mice, with a peak during the early rest span (P=0.024 and P<0.001, respectively). BAX varied fivefold during the 24 h span with a major peak occurring near mid-activity (P=0.007). The mean mRNAs of mPer2, mClock, and mBmal1 varied twofold to threefold over the 24 h, with high values during the activity span (P<0.05). In the tumor, the circadian organization in cell-cycle phase distribution was shifted and BCL2 rhythm was ablated. Conversely, a molecular circadian clock likely regulated BCL-2 and BAX expression in the bone marrow, increasing cellular protection against apoptosis during the rest span.

ARNTL Transcription Factors↗
PubMed1976

[Bone marrow's histopathology in tuberculosis of the hematopoietic organs (author's transl)].

Through the reports of the literature, the histopathologic lesions of the bone marrow in tuberculosis of the hematopoietic organs are analysed. Whether the lesions are typical or not, it is necessary to have a bacteriologic proof of tuberculosis. Near these lesions, bone marrow's cellularity is often greater or lower, reticular fibers are often increased and sometimes reticulin fibrosis is marked. Other necrosis and granulomatous inflammation are discussed. Contribution of macrophage is described. The relationships between tuberculosis and hematologic diseases are reported.

Bone Marrow↗
PubMed1995

Characteristics of peripheral blood monocytes and bone marrow macrophages from rats treated with mitomycin C, 5-fluorouracil or phenylhydrazine.

We examined the cellular functional changes in bone marrow macrophages obtained from rats treated intraperitoneally with mitomycin C (MMC), subcutaneously with 5-fluorouracil (5FU) or intraperitoneally with phenylhydrazine (PHZ) for 7 days to define the mechanisms of toxicity in the bone marrow. The functions of peripheral blood monocytes and bone marrow macrophages were also compared. MMC and PHZ caused anemia, and MMC and 5FU leukocytopenia. Changes in nucleated cell counts were different among the three drug-treatment groups, and MMC- and PHZ-treated bone marrow macrophages were activated for responsiveness to interleukin-1 (IL-1), and their migration activity, intracellular components and enzyme activities were increased. On the other hand, cellular functions of bone marrow macrophages of rats treated with 5FU were suppressed.

Animals↗
PubMed1992

The role of interleukin 6 in megakaryocyte formation, megakaryocyte development and platelet production.

Megakaryocytopoiesis is the cellular amplification and differentiation of precursors into immature megakaryocytes, and the cytoplasmic maturation of these megakaryocytes, a process terminating in the release of platelets into the circulation. Interleukin 6 (IL-6) stimulates megakaryocytopoiesis in the bone marrow, increasing platelet numbers in the circulation. IL-6 alone is poorly active on the growth of stem cell populations, but acts in synergy with stem cell factor (c-kit ligand) to expand the committed myeloid progenitor compartments but not the megakaryocyte progenitors. IL-6 has a direct action on megakaryocyte progenitors but only in synergy with low doses of interleukin 3 (IL-3), increasing the number of immature megakaryocytes and enhancing the processes of development into mature megakaryocytes. IL-6 is about 10 times more active on megakaryocytes than on megakaryocyte progenitors in cell culture. It is active alone and will stimulate increases in cell size and DNA content. IL-6 does not appear to stimulate the process of platelet release. IL-6 is found in bone marrow, in both macrophage subsets and megakaryocytes, indicating that it may be an important physiological regulator of both paracrinal (microenvironmental) and autocrinal mechanisms controlling megakaryocyte development in bone marrow.

Animals↗
PubMed1988

Enzymic programs of rat bone marrow and the impact of acivicin and tiazofurin.

The in vivo actions of two antimetabolites, acivicin (NSC-163501) and tiazofurin (NSC-286193), were examined on the enzymic programs of rat bone marrow. From the bone marrow of the femurs, 100,000 g supernatant fractions were prepared; enzymic activities were measured by isotopic assays, and cellularity was determined. In the normal bone marrow, the specific activities of pyrimidine de novo synthetic enzymes, CDP reductase, dTMP synthase, CTP synthase, carbamoyl-phosphate synthase II (synthase II), orotidine 5'-phosphate decarboxylase and aspartate carbamoyltransferase, were 1, 2.7, 5, 10, 63 and 601 nmol/hr/mg protein, respectively, whereas those of the salvage enzymes, deoxycytidine, thymidine, cytidine and uridine kinases were 3, 43, 149, and 367 nmol/hr/mg protein, respectively. In purine biosynthesis, the activities of the de novo synthetic enzymes, IMP dehydrogenase, formylglycinamidine ribonucleotide (FGAM) synthase, GMP synthase, amidophosphoribosyl-transferase (AT) and adenylosuccinate synthase were 16, 8, 107, 78 and 124 nmol/hr/mg protein, respectively, and those of the salvage enzymes, adenine, hypoxanthine and guanine phosphoribosyl-transferases, were 340, 407, and 1018 nmol/hr/mg protein, respectively. The sequence of events was elucidated after a single i.p. injection of acivicin (5 mg/kg) or tiazofurin (200 mg/kg). Within 2 hr after acivicin injection, CTP, GMP and FGAM synthases lost 85-90%, while AT and synthase II lost 50 and 80%, respectively, of their activities. The activities rose to near normal range by 72-96 hr. The bone marrow cellularity decreased, reaching a nadir at 24 and 48 hr, and returning to normal range by 72 and 92 hr; thymidine kinase activity followed a similar pattern. Tiazofurin injection depressed IMP dehydrogenase activity to 20% by 2 hr with a rebound to normal range by 48 and 72 hr. The cellularity decreased more slowly, reaching its lowest point at 24 hr and returning to normal range at 72 hr. For acivicin the marked depletion of the activities of the glutamine-utilizing enzymes and for tiazofurin that of IMP dehydrogenase might account, in part at least, for the bone marrow toxicity of these antimetabolites. Because of the presence in the bone marrow of high activities of purine and pyrimidine salvage enzymes, it should be possible to design methods utilizing nucleosides and nucleobases to protect the bone marrow from the action of antimetabolites.

Animals↗
PubMed1992

[Hematopoietic precursor cells in radiation chimeras restored by bone marrow from thymectomized mice].

Radioprotective capacity of bone marrow CFUs of adult thymectomized mice was studied. Lethally irradiated mice were inoculated with bone marrow of mice thymectomized 8-11 months before. The colony forming capacity and proliferative rate of CFUs were studied 1-7.5 months after obtaining the radiation chimeras. It has been shown that proliferative capacity of bone marrow of adult thymectomized mice was reduced in comparison with that of normal animals. It is related to the decrease (4-fold) of the proliferative rate of bone marrow of thymectomized mice which was inoculated into lethally irradiated recipients 1 month before. We also found that the content of CFUs in bone of those chimeras was reduced later--after 7.5 months. In this period (1-7.5 months) the cellularity of bone marrow did not change.

Animals↗