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Bone marrow granulocyte reserve in chronic benign idiopathic neutropenia.

Fifteen patients with chronic benign idiopathic neutropenia (CBIN) with neutrophil counts less than 1.0 X 10(9)/1 have been studied. The mean age was 33 years (range 23-50) comprising 11 females and 4 males. Bone marrow cellularity was normal except in two patients who showed slight reduction and one who had a slight increase. Bone marrow differential counts were normal apart from a small increase in the percentage of promyelocytes and reduction in the myeloid/erythroid ratio in some patients. Peripheral blood counts showed no 'compensatory' monocytosis. Epinephrine stimulation tests showed no evidence of excess neutrophil margination. After endotoxin administration there was a one- to two-fold increase in neutrophil counts in three patients, a three-fold increase in three patients and a greater than four-fold increase in the remaining nine patients. The findings suggest that the benign course of CBIN is not due to excess neutrophil margination nor to compensatory monocytosis, but that at least one mechanism includes a functionally adequate release of neutrophils to the peripheral blood under conditions of stress with subnormal delivery of neutrophils under basal conditions. The variability in responses to endotoxin suggests that CBIN is not entirely homogeneous with respect to mechanism. The findings of relatively normal bone marrow cellularity and differential counts and a normal or substantial neutrophil response to endotoxin appear characteristic. They may help predict a benign clinical course in neutropenic patients and assist diagnosis of the CBIN variant of idiopathic neutropenia.

Adult↗

Studies on the myelosuppressive activity of doxorubicin entrapped in liposomes.

The myelosuppressive activity of doxorubicin encapsulated in liposomes of differing lipid composition and size was quantified in mice by measurement of changes in spleen weight, peripheral white blood cells (WBC), and bone marrow nucleated cells. Following i.v. administration of free doxorubicin at a dose of 20 mg/kg, a 90% reduction in marrow cellularity was observed on day 3. The marrow nucleated cell count was similar to control values by day 7. Administration of an equivalent dose of doxorubicin that was encapsulated in large (diameter, approximately 1.0 microns) egg phosphatidylcholine/cholesterol (EPC/Chol)(molar ratio, 55:45) liposomes induced an 80% reduction in bone marrow cellularity that lasted for periods of greater than 7 days. Similar results were obtained following administration of large (1.0 microns) liposomal doxorubicin systems formulated with distearoylphosphatidylcholine/cholesterol (DSPC/Chol) (molar ratio 55:45). In contrast, liposomal doxorubicin prepared using small (diameter, approximately 0.1 micron) DSPC/Chol liposomes induced only a 40% reduction (day 3) in bone marrow cellularity, which returned to control values by day 7. Other indicators of doxorubicin-mediated myelosuppressive activity (spleen weight loss and peripheral leukopenia) correlated well with changes observed in marrow cellularity. An exception to this, however, was observed in animals treated with small (0.1 -micron) DSPC/Chol Liposomal doxorubicin, which displayed peripheral leukopenia for periods of greater than 14 days. This extended leukopenia was not observed following administration of small (0.1 -micron) EPC/Chol liposomal doxorubicin. Marrow-associated liposomal lipid and doxorubicin were quantified to determine if the extent of doxorubicin-mediated myeloid toxicity could be correlated to changes in biodistribution of the entrapped drug. It was demonstrated that 10-20 times more doxorubicin is delivered to the bone marrow when the drug is given encapsulated in large liposomes than when it is associated with small liposomes. These data are useful in defining characteristics of liposomal preparations that modulate the myelosuppressive behaviour of entrapped antineoplastic agents.

Animals↗

[Hypocellular acute leukemia].

To establish diagnostic criteria for hypocellular acute leukemia (HL) 32 cases (mean age 67) with 40% or less bone marrow cellularity were analysed and compared with 40 cases of MDS, 27 cases of AML in the elderly (60 > or = ) and 39 cases of AML in the young (60 <). The mean bone marrow cellularity was 30% in HL, 85% in MDS, 87% in elderly AML and 95% in young AML, respectively. Thus hypocellularity was evident in HL. Blast % in bone marrow of HL patients was 17-70% in all nucleated cells including lymphocytes (ANC), 36-93% in non-lymphocytic cells (NLC) and 50% or more in all cases in non-erythroid/non-lymphocytic cells (NENLC). Thus maturation arrest of blast cells was evident in HL, which corresponds to that of overt AML. Out of 20 cases treated with low-dose ara-C 13 cases (65%) achieved complete remission, but most of them relapsed early by manifesting hypocellular bone marrow again. In conclusion HL is a distinct clinical subtype of AML in the elderly, which can be clearly defined by 40% or less cellularity and 30% or more blasts in bone marrow.

Acute Disease↗

Effects of cadmium on haemopoiesis in irradiated and non-irradiated mice: 1. Relationship to the number of myeloid progenitor cells.

The effects of single subcutaneous injection of cadmium chloride on haemopoiesis in normal (non-irradiated) or irradiated mice were investigated. Cadmium doses used ranged from 1-8 mg/kg body weight Twenty-four hours after treatment with cadmium (doses from 3 to 8 mg/kg) there were no significant changes in bone marrow cellularity and the granulocyte-macrophage progenitor cell (GM-CFC) number per femur in non-irradiated female ICR mice. Similarly, during the 30-day postinjection period bone marrow cellularity and marrow GM-CFC number in mice treated with a cadmium dose of 5 mg/kg were not significantly different from the control values. Cadmium significantly reduced the lethal effects of gamma rays. In addition, increasing the doses of cadmium administered 24 h prior to sublethal irradiation increased the number of endogenous haemopoietic stem cells (endoCFU-S) in a concentration-dependent manner. Pretreatment with cadmium also decreased the radiation damage to endoCFU-S and haemopoietic progenitor cells committed to granulocyte/macrophage development (GM-CFC). The survival of stem cells was higher and the regeneration of cellularity and GM-CFC of irradiated bone marrow was accelerated in mice pretreated with 5 mg Cd/kg body weight in comparison with saline-injected mice.

Animals↗

Fibre content and cellularity of the bone marrow of the iliac crest, vertebral column and sternum in chronic myeloproliferative disorders.

Heterogeneous content of fibres and haematopoesis within the bone marrow may affect diagnosis and staging in chronic myeloproliferative disorders (CMPDs). To evaluate their distribution, we conducted a post mortem histomorphometric study of 22 patients with CMPD in chronic phases. In bone marrow specimens from the anterior and posterior iliac crest (right and left of each), the sternum, the 7th thoracic and the 3rd lumbar vertebra, the argyrophil fibres were counted using the line intersection method and the cellular and fatty bone marrow using the point count method. Statistical analysis was performed by direct comparison of the sites. The distribution of fibres was almost homogeneous in the patients with low fibre content, revealing a random diversity in more advanced stages of marrow fibrosis. 1/22 patient had no fibre increase in one specimen of the iliac crest and overt myelofibrosis in the other sites. 1/22 patient had myelofibrosis in two sites of the iliac crest and no fibre increase in vertebral column and sternum. The bone marrow cellularity was almost homogeneously increased in all patients. Myelofibrosis proved to be a generalised process with heterogeneous grades of severity in different regions of the bone marrow in CMPDs. No topographical bias was found. In contrast to the homogeneous increase of the bone marrow cellularity the topographical heterogeneity of the fibre content may limit the representativity of single bone marrow biopsies in patients with CMPDs.

Adult↗

Immunopotentiating activity of abrin, a lectin from Abrus precatorius Linn.

A non-toxic dose of abrin, (1.25 microg/kg body wt) consecutively for five days in normal mice stimulated specific humoral responses. A noticeable increase was observed in total leucocyte count, lymphocytosis, weights of spleen and thymus, circulating antibody titre, antibody forming cells, bone marrow cellularity and alpha-esterase positive bone marrow cells. The results suggest that abrin can potentiate the humoral immune response of the host.

Abrin↗

Administration of the bacterial extract Broncho-Vaxom enhances radiation recovery and myelopoietic regeneration.

In the present study, we show that the bacterial extract Broncho-Vaxom (BV, 500 micrograms/mouse; free of endotoxin) has radiation recovery activity when administered i.p. 24 h before sublethal irradiation. In the postirradiation period (5-12 days), pretreatment of mice with BV induced significantly increased bone marrow cellularity and accelerated myelopoietic regeneration (committed progenitor granulocyte-macrophage colony-forming cells; GM-CFC) in the bone marrow compared with saline-treated controls. The earlier hemopoietic recovery in BV-injected mice was not associated with an increase in the number of bone marrow GM-CFC and CFU-S (colony-forming units-spleen) within 24 h after injection. Simultaneously, a significant diminution in bone marrow cellularity occurred. In addition, the percentage of both GM-CFC and CFU-S in the S-phase of the cell cycle was significantly increased 24 h after a single treatment. In our experiments colony stimulating activity (CSA) in the serum of treated mice was not observed within 24 h after injection. Administration of BV 24 h prior to lethal irradiation, resulted in an increase in the number of surviving mice. Combined administration of BV (24 h) and indomethacin (24 h and 3 h) to mice, prior to irradiation, caused an additional radioprotective effect. These results demonstrate that BV stimulates myelopoietic regeneration and suggest a mechanism by which this treatment protects mice from otherwise lethal irradiation.

Adjuvants, Immunologic↗

Bone marrow cells of mice under the influence of low lead doses.

The influence of lead on the number of nucleated bone marrow cells and pluripotent stem cells in femur bone marrow was investigated in mice. It was found that neither a single nor 10 days' oral administration of lead at doses of 10-1,000 micrograms PbAc/kg b.w. led to any change in the bone marrow cellularity. After 1 month's oral administration of 1,000 micrograms PbAc/kg b.w., on the other hand, there was a significant reduction of nucleated bone marrow cells and pluripotent stem cells in the femur bone marrow of up to 20% in comparison with controls. Within 14 days after completing the lead administration, however, normal values in the bone marrow cellularity were found again. Three months' oral administration of lead at doses of 100 and 1,000 micrograms PbAc/kg b.w. led to a significant reduction in nucleated bone marrow cells and pluripotent stem cells in the femur bone marrow of up to 28% compared with the controls. In contrast to the experiments with 1 month's oral administration of lead the regeneration of the bone marrow was not completed 14 days after completing the administration of the lead. These results show that lead at doses above 100 micrograms PbAc/kg b.w. causes on long-term administration a clear reduction of the nucleated bone marrow cells combined with a depressed regeneration ability of the pluripotent stem cells.

Animals↗

Chemoprotective activity of an extract of Phyllanthus amarus against cyclophosphamide induced toxicity in mice.

The effect of 75% methanolic extract of the plant Phyllanthus amarus (P. amarus) was studied against cyclophosphamide (CTX) induced toxicity in mice. Administration of CTX (25 mg/kg b.wt, i.p.) for 14 days produced significant myelosuppression as seen from the decreased WBC count and bone marrow cellularity. Administration of P. amarus extract at doses 250 and 750 mg/kg b.wt significantly reduced the myelosuppression and improved the WBC count, bone marrow cellularity as well as the number of maturing monocytes. CTX treatment also reduced the activity of glutathione system and increased the activity of phase I enzyme that metabolize CTX to its toxic side products. P. amarus administration was found to decrease the activity of phase I enzyme. Administration of P. amarus also increased the cellular glutathione (GSH) and glutathione-S-transferase (GST), thereby decreasing the effect of toxic metabolites of CTX on the cells. Administration of P. amarus did not reduce the tumor reducing activity of CTX. In fact, there was a synergistic action of CTX and P. amarus in reducing the solid tumors in mice. Results indicated that administration of P. amarus can significantly reduce the toxic side effects of CTX and is not interfering with the antitumor efficiency of CTX.

Animals↗

Evaluation of tissue disposition, myelopoietic, and immunologic responses in mice after long-term exposure to nickel sulfate in the drinking water.

Female B6C3F1 mice were exposed to graded doses of nickel sulfate to determine a threshold response for myelotoxicity and immunotoxicity, and to identify which of the populations of lymphoreticular cells were most sensitive to the toxic effects of nickel. Animals were given free access to the chemical in the drinking water at 0, 1, 5, or 10 g/l for 180 d. Water consumption, blood and tissue nickel concentrations, body and organ weights, histopathology, immune responses, bone marrow cellularity and proliferation, and cellular enzyme activities were evaluated. There was no mortality. Mice in the 5-g/l and 10-g/l dose groups drank less water than controls; the responses measured in the 10-g/l group may have been due to a combination of dehydration and chemical toxicity. Decreases in body and organ weights were confined to mice in the 10-g/l dose group, except for the dose-related reductions in thymus weights. Blood nickel was measured at 4, 8, 16, and 23 wk of exposure. The mean blood nickel values showed increases between 4 and 8 wk that were proportional to time and dose; thereafter there was no substantial increase in blood nickel in any of the dose groups, except for an increase in the mean blood concentration in the 10-g/l group at 23 wk. The kidney was the major organ of nickel accumulation. The primary toxic effects of nickel sulfate were expressed in the myeloid system. There were dose-related decreases in bone marrow cellularity, and in granulocyte-macrophage and pluripotent stem-cell proliferative responses. In unfractionated bone marrow cells glucose-6-phosphate dehydrogenase enzyme activity from the hexose monophosphate shunt was more sensitive to nickel sulfate than were representative glycolytic or Krebs cycle enzymes, with 25-35% maximum inhibition at 5 g/l and 10 g/l. Aliquots of bone marrow cells were separated into enriched bands of lymphocytes, granulocyte-macrophages, and erythrocytes; enzyme inhibition that occurred in unfractionated bone marrow cell aliquots was only expressed after cell separation in the enriched granulocyte-macrophage cell population, suggesting that these committed stem cells were a primary target of nickel sulfate toxicity. There was one example of systemic immunotoxicity, reduction in the lymphoproliferative response to lipopolysaccharide, and it was regarded as secondary to the primary effect of nickel sulfate on the myeloid system, since this was the only significant change among a panel of seven immune parameters that were evaluated.

Animals↗

The effect of two glucan carboxymethyl derivatives with various substitution degrees on cyclophosphamide immunosuppression in mice.

Carboxymethylglucan (CMG) in two different degrees of substitution of carboxymethyl groups (0.56 and 0.89) was administered to cyclophosphamide (CY) treated (200 mg/kg) C57B1/6 mice in three doses (10, 50, and 100 mg/kg) 24 h after CY. The influence of CMG administration on the cell suppression caused by CY was observed in the subsequent days. The cellularity of spleen, bone marrow and peripheral blood was quantified on days 2, 5, 8, 11, 15, and 21 after CY treatment. CY treated mice developed a significant decrease in peripheral blood cell counts, and spleen and bone marrow cellularity during the initial 5 days, followed by recovery in the next 15 days, with an overshoot reaction in spleen cellularity and erythrocyte levels. The initial cellularity depression and the following recovery was modified by both carboxymethyl derivatives of glucan. Cyclophosphamide treated mice exhibited less pronounced immunosuppression and more rapid hematopoietic recovery when administered with CMG, although this reaction was only of a modest degree. The effects were not dose dependent and the differences between the two glucans were not significant.

Animals↗

Androgen therapy for anemia in renal failure.

Forty-six patients marked renal failure and anemia underwent androgen therapy (up to 500 mg/week), as well as intravenous injections of iron preparations (600-800 mg/month) and hemotransfusions (2500 ml/month) for 4 month. Ferrokinetic indices, total marrow cellularity and erythrocyte and hemoglobin levels were determined at the end of therapy. Androgens were responsible for the increase in Hb level by 1.2 g%. High total bone marrow cellularity was observed in these patients (27.4 +/- 3.2 X 10(9) cells/kg, compared with 14.1 +/- 1.4 X 10(9) cells/kg in normal individuals). Hemotransfusions resulted in a decrease of Hb level by 1.3g% and total bone marrow cellularity to 8.3 +/- 2.2 X 10(9) cells/kg. Ferrokinetic indices became poor. The data obtained are discussed.

Adult↗

7,12-Dimethylbenz[a]anthracene-induced bone marrow toxicity is p53-dependent.

Polycyclic aromatic hydrocarbons (PAHs) are known immunotoxins and carcinogens. Our laboratory and others have demonstrated that metabolism of these compounds by CYP1B1 is required for carcinogenicity and immunotoxicity to occur. Previously, our laboratory reported significantly decreased bone marrow cellularity in mice following 7,12-dimethlybenz[a]anthracene (DMBA) administration. In addition, we have observed that DMBA causes apoptosis via activation of both caspase-8 and -9 in pre-B cells co-cultured with bone marrow stromal cells in vitro. In this study, we investigated the importance of the p53 protein in the bone marrow response to DMBA. Through the use of p53 gene knockout mice, we demonstrated that the effect of DMBA on bone marrow cellularity is p53-dependent. In addition, apoptosis of primary cultures of progenitor B cells cultured with bone marrow stromal cells and DMBA is also p53-dependent. The results of this study provide evidence for the importance of p53 in the signaling pathways by which PAHs cause immunotoxicity.

9,10-Dimethyl-1,2-benzanthracene↗

Bone marrow biopsy in monoclonal gammopathies: correlations between pathological findings and clinical data. The Cooperative Group for Study and Treatment of Multiple Myeloma.

Between January 1987 and October 1989, 561 consecutive untreated patients with monoclonal gammopathy of undetermined clinical importance (MGUS) (n = 295) or with multiple myeloma (n = 266) were evaluated in a multicentre trial. Both bone marrow biopsy and aspiration (performed at different anatomical sites) were required at presentation. Bone marrow biopsy data indicated that changes in bone marrow composition from MGUS to early multiple myeloma and to advanced multiple myeloma followed a precise pattern, including an increased percentage of bone marrow plasma cells (BMPC%), a shift from plasmocytic to plasmoblastic cytology, an increase in bone marrow cellularity and fibrosis, a change in bone marrow infiltration (becoming diffuse rather than interstitial), a decrease in residual haemopoiesis and an increase in osteoclasts. In multiple myeloma the BMPC% of biopsy specimens and aspirate were closely related, although in 5% of cases the difference between the two values was greater than 20%. Some histological features were remarkably associated with each other. For example, BMPC% was higher in cases with plasmoblastic cytology, heavy fibrosis, or reduced residual haemopoiesis. Anaemia was the clinical characteristic most influenced by bone marrow histology. The BMPC% was the only histological variable which affected the greatest number of clinical and laboratory characteristics, including, besides haemoglobin concentration, erythrocyte sedimentation rate, radiographic skeletal bone disease, and serum concentrations of monoclonal component, calcium, beta 2-microglobulin and thymidine kinase activity. These data indicate that comparative bone marrow histology in monoclonal gammopathies has clinical importance.

Aged↗

[Radiomodifying effect of serotonin on the cells of the hematopoietic system].

Mice were injected with serotonin (0.5 and 2.0 mg/mouse) 15 min before and immediately after irradiation (4.5 and 7.0 Gy). With the preparation administered prior to irradiation one could observe an increase in the quantity of CFUc survived (endo- and exo-test), acceleration of restoration of cellularity (in the bone marrow) and growth of the spleen weight. The acceleration of restoration of the bone-marrow cellularity was also noted when serotonin was administered after irradiation. The post-irradiation effect of serotonin was also detected in cells in vitro. It is concluded that the radiomodifying effect of serotonin on the haemopoietic system is due to a decrease in the number of damaged cells and increase in the rate of reproduction of intact cells.

Animals↗

[Morphological observation of a process of bone marrow hypoplastic formation in a case of aplastic anemia].

Bone marrow morphological change was consecutively analyzed form the disease onset to the formation of bone marrow aplasia in a patient with post-hepatitic aplastic anemia. In this case, the mean bone marrow cellularity and absolute numbers of erythroids and megakaryocytes were continuously higher than those in normal subjects for 3 weeks after the appearance of peripheral pancytopenia. During this stage, administration of recombinant human granulocyte colony-stimulating factor (G-CSF) improved marrow myeloid hypoplasia and peripheral neutropenia. During the period in which the marrow cellularity transformed from hyperplasia to hypoplasia, the bone marrow showed a mixture of hyper-, normo- and hypocellular portions, and the decrease in the megakaryocytes was the faster than myeloid and erythroid cells. These findings indicate that (1) ineffective hematopoiesis might be present in the early stage of the disease, (2) G-CSF responsive granulocytic precursors remained during the early stage of the disease, and (3) the marrow aplasia progressed in the manner of aplastic nest formation during the period in which the marrow cellularity declined to hypoplasia. We experienced another case of aplastic anemia showing the same progress of bone marrow findings and speculated that this might be one of the ways of the progression of bone marrow hypoplastic formation in aplastic anemia.

Adolescent↗

The effect of a Titanocene Dichloride derivative, Ti IV (C5H5)(2) NCS(2), on the haematopoietic response of Ehrlich tumour-bearing mice.

The effects of the [Ti IV (C(5)H(5))(2) NCS(2)] metallocene (BCDT), a Titanocene Dichloride derivative, on the growth and differentiation of granulocyte-macrophage progenitor cells [colony-forming unit-granulocyte-macrophage (CFU-GM)] and bone marrow cellularity in normal and Ehrlich ascites tumour-bearing mice were studied. As expected for the Ehrlich ascites tumour-model, concomitant myelosuppression, increased number of spleen CFU-GM and changes in bone marrow cellularity were observed. The treatment of Ehrlich ascites tumour-bearing mice with BCDT (10-30 mg/kg/day) produced a dose-dependent increase in myelopoiesis, a reduction in splenic colonies and a restoration in the total and differential marrow cell counts. We also observed an increase in CFU-GM number when bone marrow cells obtained from normal mice were incubated in vitro with serum from normal mice treated with BCDT. In addition, BCDT prolonged, in a dose-dependent manner, the survival of mice inoculated with Ehrlich ascites tumour. Although it has been previously reported that substitutions in the two halides of the titanocene do not interfere with antitumoural effect, our results with BCDT demonstrated a reduction in antitumour efficacy when compared to previous results with the original titanocene produced in our laboratory.

Animals↗