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Hematopoietic effects in mice exposed to arsine gas.

Arsine gas is a potent hemolytic agent. Concern about semiconductor workers prompted an in-depth study of arsine at the National Institute of Environmental Health Sciences to determine the hematopoietic effects of prolonged exposure to this gas. Female B6C3F1 mice were exposed by inhalation to 0, 0.5, 2.5, and 5 ppm arsine, 6 hr/day for 14 days. Body weights of exposed mice were comparable to those of controls, but a marked, concentration-related splenomegaly was observed. Higher level arsine exposure produced statistically significant decreases in red blood cells, hematocrit and hemoglobin, with increases in white blood cell counts and mean corpuscular volume of red blood cells. Erythropoiesis as measured by quantitation of erythroid precursors in culture revealed a marrow reduction of colony-forming unit erythroids/femur cells for all treated groups on Day 3 postexposure and only at the 5 ppm dose group on 24 days postexposure, while splenic erythropoiesis increased at higher concentrations of arsine. There was no alteration in bone marrow cellularity and a less significant effect on granulocyte-macrophage progenitors. A 12-week study of arsine at 0, 0.025, 0.5, and 2.5 ppm (6 hr/day) by inhalation showed similar effects on hematopoiesis in mice. In conclusion, arsine exposure at low concentrations produces a stress on the hematopoietic system characterized by hemolysis, which persists for a prolonged period following exposure.

Animals↗

Peripheral blood stem cells collected in very early remission produce rapid and sustained autologous haemopoietic reconstitution in acute non-lymphoblastic leukaemia.

Haemopoietic reconstitution was achieved in a patient with acute non-lymphoblastic leukaemia (ANLL) in relapse who was autografted with blood-derived stem cells collected during very early remission. The patient received a myeloid progenitor cell dose of 230 x 10(4) CFU-GM/kg body weight. Engraftment was evident in the bone marrow 7 days post-graft. Normal neutrophil and platelet counts were attained by day 14 and blood counts remained normal thereafter. An overshoot in peripheral blood haemopoietic progenitor levels occurred at the end of the second week, presumably the progeny of a family of early progenitor cells. The completeness of haemopoietic reconstitution is further illustrated by the satisfactory nucleated cell and myeloid progenitor cell yield when a bone marrow harvest was performed 4 1/2 months post-graft. Seven months post-graft, the patient remained in complete remission with normal blood counts and bone marrow cellularity, although haemopoietic progenitor levels were slightly reduced. The rapid recovery minimises aplasia-related risks and suggests that such autografting can be carried out safely in first remission. We propose that autografting using very early remission blood cells is a new therapeutic option for patients with acute ANLL.

Adolescent↗

[Bone marrow granulomas].

PURPOSE: To evaluate bone marrow granulomatous lesions in order to establish their etiology. MATERIAL AND METHODS: 2,250 bone marrow biopsies were studied during the period of March 1983-March 1991. Granulomas and/or granulomatous lesions were found in 24 of them (1.06%). A correlation between histological characteristics, special stains: PAS, Ziehl Neelsen and Grocott and cultures were done. Immunohistochemistry was done to evaluate B or T cell-lineage in 4 patients. RESULTS: The 24 patients were biopsied because of the clinical diagnosis of haematological and non-haematological neoplasias, infections, AIDS, sarcoidosis and fever of unknown origin. Bone marrow cellularity ranged from 20% to 75% (M: 49.8%). Myeloid cells were increased in 54% of the cases. The number of granulomas ranged from 1 to 19 (M: 3.9). The epithelioid cells were the predominant component in 66% of the cases. Of the 7 patients with non-Hodgkin's lymphoma, 4 had lymphomatous involvement with granulomatous pattern. These cases showed predominance of lymphoid cells and vessels in addition to epithelioid cells. CONCLUSION: We consider that in order to establish a relationship between infection and granuloma, the identification of a microorganism through a culture is a more reliable test. We couldn't find any morphological characteristic which allowed an etiologic diagnosis of bone marrow granulomas. In case of lesions with a great lymphocytic and vascular proliferation plus the presence of epithelioid cells and fibrosis, NHL with bone marrow involvement with a granulomatous pattern should be strongly considered.

Acquired Immunodeficiency Syndrome↗

Minimally differentiated acute leukemia.

We have studied 35 adult patients with morphologically undifferentiated peroxidase-negative acute leukemia that failed to meet the criteria for acute lymphoblastic leukemia and compared them to patients with FAB M1-M7 seen by the same physicians. The diagnosis of minimally differentiated acute leukemia (MD-AL) was associated with a higher incidence of prior hematologic disease, lower WBC, fewer blood blasts, lower marrow cellularity and a tendency towards older age. Of all patients treated with AML since January 1983, those with MD-AL were less likely to get a complete remission than those with other subtypes (35 vs 64%, p = 0.03). Treatment failure was usually due to resistant disease. Analysis of outcome as a function of drugs used during induction therapy showed an advantage for regimens containing vincristine and prednisone. The leukemic blast cells of nine patients were immunophenotyped for myeloid, lymphoid and megakaryoblast/platelet antigens. Although there were too few for a full statistical analysis as was applied to the larger group of 35 patients with MD-AL, these patients had a lower bone marrow cellularity as compared to FAB M1-M7 and a low remission rate. Eight of these were found to have positive myeloid markers and met the criteria for FAB M0. We conclude that patients with MD-AL form a distinct group with characteristic presenting features and a low response rate. Outcome data suggest that vincristine and prednisone should be included in experimental induction programs.

Acute Disease↗

Increased telomerase activity is associated with shorter survival in patients with chronic phase chronic myeloid leukemia.

BACKGROUND: Significantly elevated telomerase activity (TA) has been found in samples from patients with many malignant hematologic diseases. However, the impact of elevated TA on the course of patients with chronic phase chronic myeloid leukemia (CP-CML) is unknown. METHODS: Using a modified polymerase chain reaction-based telomeric repeat amplification protocol assay, the authors measured TA in bone marrow samples from 93 patients with CP-CML and correlated it with patient characteristics and survival. TA also was measured in bone marrow samples from 29 patients with accelerated/blastic phase CML. RESULTS: Patients with accelerated/blastic phase CML were found to have somewhat higher levels of TA compared with patients with CP-CML (P = 0.07). Among patients with CP-CML, those with high TA progressed to advanced stages of disease sooner (P = 0.05) and had a significantly shorter survival (P = 0.04) than patients with low TA. No correlation was found between TA and patient age, hemoglobin, platelet and leukocyte counts, percentage of peripheral or bone marrow blasts or basophils, or bone marrow cellularity. On multivariate analysis, high TA retained its significance as a factor associated with shorter patient survival (P = 0.02). CONCLUSIONS: The current study data suggest that TA plays a role in the propagation of CP-CML and that the potential of telomerase inhibitors in patients with CML should be explored, even in those with early phase disease.

Aged↗

Alterations in hematopoietic responses in B6C3F1 mice caused by drinking a mixture of 25 groundwater contaminants.

Myelotoxicity parameters were monitored in female B6C3F1 mice exposed to 0, 1, 5, and 10% of a chemical mixture stock in drinking water for 2.5 to 31.5 weeks. The mixture consisted of 25 groundwater contaminants frequently found near toxic waste dumps, as determined by U.S. Environmental Protection Agency (EPA) surveys. Water consumption, body and organ weights, and hematological and histopathological examinations were conducted. No animals developed overt signs of toxicity after 2.5 weeks of treatment. No significant effect on bone marrow cellularity was observed after 2.5, 15.5, or 31.5 weeks of exposure; however, mice exposed to 5% or higher concentrations of the chemical mixture stock solution for 15.5 weeks showed significant suppression of granulocyte-macrophage progenitor cells (CFU-GM) and erythroid precursors (CFU-E) with no changes in body weight, histopathological or hematological parameters. Decreases occurred in erythrocyte mean corpuscular volume of mice exposed to a 10% solution for 15.5 weeks and to 5 and 10% solutions following 31.5 weeks of treatment. In addition, dose-related decreases were found in body, liver, and thymus weights in the 5 and 10% solutions exposure groups after 31.5 weeks. These results suggest that bone marrow may be a sensitive indicator for long-term, low-level exposure of multiple chemicals in mice. Furthermore, long-term exposure to highly contaminated groundwater may present a subtle risk to the hematopoietic system.

Animals↗

[Bone marrow biopsy in non-Hodgkin's lymphoma: experience with 259 samples].

A study performed on 259 cases of bone-marrow biopsy specimens from 121 patients with non-Hodgkin's lymphoma (NHL) is presented after an introduction dealing with several well-known facts. Specific involvement of the initial samples was present in 53.45% of the cases. Such involvement was more frequent in stages III and IV, although no difference was found between the clinical forms, A or B. Stage progression due to positive bone-marrow biopsy occurred in 26% of the patients, and in 25% of these it was from stages 0 to II onto stage IV. Low-grade lymphomas showed higher bone-marrow involvement than high-grade ones (p = 0.025); specific involvement significantly influenced survival in only high-grade lymphomas, not in the low-grade ones, when the series was considered as a whole. Absolute compatibility between lymph-node and bone-marrow cellularity was found in 71% of the cases, and in 85% was taken into account. Six possible histological lesions are described herein, while for practical purposes, the use of 5 patterns of involvement is recommended, namely, interstitial, nodular, patchy, mixed nodular-patchy, and diffuse. Patchy pattern associated to low-grade and skin-involvement lymphomas. Although those patterns had non-significant influence on survival, this last was found when comparing diffuse pattern with the remainders; similarly, the diffuse pattern correlated with significantly lower haemoglobin rates. Finally, fibrosis was present in 58% of the positive biopsies and in 94% of the lesions, it being thus considered as a marker of bone-marrow involvement.

Bone Marrow↗

Effect of naturally occurring triterpenoids glycyrrhizic acid, ursolic acid, oleanolic acid and nomilin on the immune system.

The effect of naturally occurring triterpenoid compounds such as glycyrrhizic acid, ursolic acid, oleanolic acid, and nomilin on the immune system was studied using Balb/c mice. Intraperitoneal treatments with 5 doses of these terpenoid compounds were found to enhance the total white blood cells (WBC) count. In ursolic acid, oleanolic acid and nomilin treated animals the maximum total WBC count was observed on the 6th day, while in glycyrrhizic acid treated animals it was observed only on the 9th day after the drug treatment. In ursolic acid, oleanolic acid and nomilin treated animals the percentage of increase in the total WBC count was to 91.48 +/- 4.6%, 135.75 +/- 6.4% and 117.33 +/- 17.9% respectively. In the glycyrrhizic acid treated animals the total WBC count was increased to 114.9 +/- 18%. Bone marrow cellularity and alpha-esterase positive cells were also enhanced by the treatment with these terpenoids. Treatment with various triterpenoids along with antigen produced an enhancement in the specific antibody titre and the number of plaque forming cells (PFC) in the spleen. Triterpenoids remarkably inhibited delayed type hypersensitivity reaction (DTH). These results indicate the immunomodulatory activity of naturally occurring triterpenoids such as glycyrrhizic acid, ursolic acid, oleanolic acid and nomilin.

Animals↗

Decrease in hematopoietic stem cell domains as a delayed effect of x-irradiation.

Although the hematopoietic integrity of locally X-irradiated sites can be restored for a time even after fairly large doses, a secondary aplasia often occurs some months later. To gain further insight into this delayed effect within the framework of the stem cell regulatory domain hypothesis, we characterized the growth kinetics of spleen colony forming units (CFU-S) in WBB6FI-+/+ bone marrow transplanted into WBB6FI-W/WV mice in which one leg had been exposed to 10-30 Gy of X rays 4-5 months previously. Compared to unirradiated contralateral marrow, fewer CFU-S either reached the previously irradiated marrow or were seeded into sites that could support growth. The initial exponential growth of effectively seeded CFU-S was unchanged, but growth deceleration (inflection point) occurred at a lower level of CFU-S in marrow previously irradiated with 20-30 Gy. This change in the inflection point indicates a radiation dose-dependent decrease consistent with the decrease in bone marrow cellularity. The decrease in effective stem cell domains after 20 Gy was calculated to be about 35%. We interpret these results to reflect the highly localized nature of delayed radiation damage to the marrow microenvironment.

Animals↗

Immunological function in mice exposed to JP-8 jet fuel in utero.

Immunological parameters, host resistance, and thyroid hormones were evaluated in F1 mice exposed in utero to jet propulsion fuel-8 (JP-8). C57BL/6 pregnant dams (mated with C3H/HeJ males) were gavaged daily on gestation days 6-15 with JP-8 in a vehicle of olive oil at 0, 1000, or 2000 mg/kg. At weaning (3 weeks of age), no significant differences were observed in body, liver, spleen, or thymus weight, splenic and thymic cellularity, splenic CD4/CD8 lymphocyte subpopulations, or T-cell proliferation. Yet, lymphocytic proliferative responses to B-cell mitogens were suppressed in the 2000 mg/kg treatment group. In addition, thymic CD4-/CD8+ cells were significantly increased. By adulthood (8 weeks of age), lymphocyte proliferative responses and the alteration in thymic CD4-/CD8+ cells had returned to normal. However, splenic weight and thymic cellularity were altered, and the IgM plaque forming cell response was suppressed by 46% and 81% in the 1000 and 2000 mg/kg treatment groups, respectively. Furthermore, a 38% decrease was detected in the total T4 serum hormone level at 2000 mg/kg. In F1 adults, no significant alterations were observed in natural killer cell activity, T-cell lymphocyte proliferation, bone marrow cellularity and proliferative responses, complete blood counts, peritoneal and splenic cellularity, liver, kidney, or thymus weight, macrophage phagocytosis or nitric oxide production, splenic CD4/CD8 lymphocyte subpopulations, or total T3 serum hormone levels. Host resistance models in treated F1 adults demonstrated that immunological responses were normal after challenge with Listeria monocytogenes, but heightened susceptibility to B16F10 tumor challenge was seen at both treatment levels. This study demonstrates that prenatal exposure to JP-8 can target the developing murine fetus and result in impaired immune function and altered T4 levels in adulthood.

Administration, Oral↗

Locoregional application of 5-fluorouracil to the liver: alteration of pharmacokinetics and bone marrow toxicity in rats with thioacetamide-induced cirrhosis.

Pharmacokinetics and toxicity of 5-fluorouracil (5-FU) applied intra-arterially into the liver were studied in control and cirrhotic rats. Cirrhosis was induced by administering thioacetamide (4.5 mg/rat X day) over 6 months. 5-FU was administered into the hepatic artery at 30 mg/kg b.w. by bolus injection followed by infusion of 0.63 mg/kg X min for 40 min. Pharmacokinetics of 5-FU in plasma was studied by HPLC analysis. 5-FU induced toxicity was examined in a second group of control and cirrhotic rats at 48 and 168 hours after treatment with 5-FU (31.4 mg/kg b.w.) infused into the liver artery for 1 h. Analysis of toxicity was carried out by determining liver enzymes in plasma, bone marrow cellularity and colony forming units in vitro (CFU-C) and in vivo (CFU-S). The results indicate that during the period investigated (85 min) 5-FU was not eliminated from the plasma of cirrhotic rats in contrast to controls. The constant level of 5-FU in the plasma of cirrhotic rats induced a considerably higher myelosuppression in these animals, than the short-lived 5-FU peak in controls.

Alanine Transaminase↗

Immunomodulatory effect of some naturally occuring sulphur-containing compounds.

The immunomodulatory effects of naturally occurring sulphur compounds such as diallyl sulphide (DAS), diallyl disulphide (DADS) and allyl methyl sulphide (AMS) were studied in BALB/c mice. After treatment with five doses (20 mg/dose) of sulphur-containing compounds, the total white blood cell (WBC) count was enhanced significantly in mice. Among the sulphur compounds studied, DADS showed the maximum number of WBC (17,900 cells/mm(3)) on the 6th day, and highest antibody titre of 516 on the 12th day. Administration of DAS and DADS enhanced the weight of vital organs such as the spleen and thymus. DADS, AMS, and DAS administration could enhance the number of plaque forming cells (PFC) in the spleen of the animals. Maximum numbers of PFC (1,409 PFC/10(6) spleen cells) were observed in DADS-treated animals. Bone-marrow cellularity was also increased significantly (P<0.001) in DADS (19.3 x 10(6) cells/femur) and AMS (24.7 x 10(6) cells/femur) treated animals. The number of alpha-esterase positive cells was enhanced in DAS (1,315/4,000 cells), DADS (1,748/4,000 cells), and AMS (1,648/4,000 cells) treated animals, compared to normal animals (1,065/4,000 cells). These results are suggestive of an immunostimulating effect of sulphur compounds.

Adjuvants, Immunologic↗

Yeast-expressed granulocyte-macrophage colony-stimulating factor in cancer patients: a phase ib clinical study.

The in vivo effect of yeast-derived recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) was investigated in 29 patients with advanced malignancy in phase Ib trial. Patients were treated at six different dose levels (30-1000 micrograms/m2/day) with either daily intravenous bolus injection or 24 hours continuous infusion for 5 days or 2 weeks. Administration of rh GM-CSF resulted in a broad spectrum of dose-, route-, and schedule-dependent hematopoietic effects. Sustained infusion of rh GM-CSF elicited a maximum 17-fold average peak increase of the total white blood cell (WBC) count with mainly neutrophils, eosinophils, and monocytes accounting for this rise, and increases in bone marrow cellularity with a shift to immature myeloid elements. Elevation of lymphocytes, platelets and reticulocytes was not induced. Within one week after discontinuation of treatment the leukocytosis had disappeared. Adverse reactions encountered with rh GM-CSF seen in 65% of the patients studied were never life-threatening and always reversible. They included mild myalgias, facial flushing, low-grade fever, headache, bone discomfort, nausea, dyspnoea and transient decline of platelet counts. These results suggest that rh GM-CSF can be safely administered at the doses and schedules employed and that it can induce in vivo some of the biological effects reported in in vitro studies. Although no objective antitumour responses have been seen, the ability of rh GM-CSF to increase turnover and function of leukocytes in vivo may prevent neutropenia and infections, when GM-CSF is adjunctively added to cytotoxic cancer therapy.

Adult↗

Allogeneic blood cell transplantation following reduced-intensity conditioning is effective therapy for older patients with myelofibrosis with myeloid metaplasia.

Standard myeloablative conditioning prior to allogeneic hematopoietic stem cell (HSC) transplantation has been associated with significant toxicity in patients older than 45 years of age with myelofibrosis with myeloid metaplasia (MMM). We sought to evaluate the efficacy of a reduced-intensity conditioning regimen for allogeneic HSC transplantation in this setting. A regimen consisting of fludarabine (30 mg/m(2) intravenously daily for 5 days) and melphalan (70 mg/m(2) intravenously daily for 2 days) followed by transplantation of filgrastim-mobilized peripheral blood cells from HLA-identical siblings was administered to 4 older patients (median age, 56 years; range, 48-58 years) with advanced MMM. All patients achieved prompt neutrophil and platelet engraftment and have experienced a significant regression of splenomegaly and bone marrow fibrosis. All now have normal bone marrow cellularity. With a median follow-up of 13 months (range, 11-19 months), all 4 patients are alive with stable full-donor hematopoietic chimerism. These results support the feasibility and effectiveness of reduced-intensity conditioning prior to allogeneic HSC transplantation for older patients with advanced MMM.

Antineoplastic Combined Chemotherapy Protocols↗

[New data about oligoblastic leukemias. Apropos of an analysis of 120 cases].

We studied 120 aptients within the following condition: moderate bone marrow myeloblastosis (blast cells + promyelocytes less than or equal to 50 p. cent); quantitative and/or qualitative medullary insufficiency; preservation of maturation of the granulocytic line without a gap; WBC count less than 20 000/mm-3. Computer analysis pointed out the following results: 1. This group may be differenciated from the group of acute myeloblastic leukemias by: a) a predilection for subjects older than 60 in 73 p. cent of cases; b) male dominance in 70 p. cent of cases; c) a sharply defined improved prognosis with a median of survival of 15 months and with more than 30 p. cent of survivors for more than two years; d) death in 63 p. cent of teh cases is directly due to a complication of bone marrow insufficiency, and only in 14 p. cent of the cases is related to blastic invasion; e) a complete absence of favourable response to intensive chemotherapy. 2. There are five initial prognostic parameters which are of statistical significance: a) the hemoglobin level; b) the sedimentation rate; c) the platelet count, the level of bone marrow cellularity, and age. 3. Multidimensional analysis makes it appear that the whole population obeys a unidimensional phenomenon and depends on a common "biological axis" constituted by the five prognostic factors which are inter-related. 4. The disease nevertheless may be subdivided into three subgroups; a) High inflammatory hypoplastic forms are a very high risk group; b) The group of slowly invasive acute leukemias in young subjects is a bad or moderately poor risk group; c) The forms with a purely qualitative disorder of myelopoiesis with very low evolutive potential. These observations point to: a) The concept of OBL as an entity of a distinctive type including the preleukemic syndrome, the myelo-monocytic leukemias, and the smoldering leukemias; b) The concept of a unique and basic bone marrow disorder which embraces the OBL and the common acute myeloblastic leukemia.

Adult↗

Telomerase activity is not a prognostic factor in chronic lymphocytic leukemia.

We measured telomerase activity (TA) in bone marrow samples from 214 patients with CLL and correlated it with patients' characteristics and survival. In >50% of cases (126/214; 59%) no detectable TA was found. There was no difference in TA between previously treated (n = 153) and untreated (n = 61) patients (P = 0.4), or patients with various Rai (0-IV) stages (P = 0.85). TA correlated significantly with white blood cell and lymphocyte count (P = 0.02 and 0.01, respectively) but not with bone marrow cellularity, beta2-microglobulin (beta2M), or other patient characteristics. Patients who had no TA had slightly lower beta2M and lower lymphocyte counts (P = 0.5 and 0.04, respectively) as compared with patients with detectable TA. However, there was no correlation between TA and survival. This data suggests that TA may not play a significant role in the clinical behavior of CLL.

Adult↗

In vitro inhibition of murine hematopoietic progenitors and stromal cells by vinorelbine.

Hematopoietic progenitor colony assays were used to establish the effects of the vinca alkaloid vinorelbine (VRB) on murine bone marrow. The in vitro growth of colony-forming units-granulocyte/macrophage (CFU-GM), burst forming units-erythroid (BFU-E) and colony-forming units-mix (CFU-mix) was dose-dependently inhibited by VRB. The highest dose assayed (0.02 microg/ml) suppressed all of the different progenitor cells by 100%. A comparison of the dose-response curves showed that CFU-GM, BFU-E, and CFU-mix exhibited similar-patterns of sensitivity to the cytotoxic action of VRB. Long-term bone marrow cultures have provided a valuable in vitro model for studying the role of the microenvironment of bone marrow. Cellularity of stromal layers was reduced with increasing doses of VRB. The appearance of these layers was altered minimally with the lowest dose used; a gradual loss of cellularity was seen in cultures exposed to 0.05 and 0.075 microg/ml; and a marked loss at the dose of 0.1 microg/ml. Our results show that VRB has an important effect on hematopoietic progenitors at the highest dose tested, while the stromal cells were not affected at a similar dose (0.025 microg/ml), suggesting that the stroma is more resistant to this drug.

Animals↗

Haematological, bone marrow and clinical chemical changes in neonatal foals given canine recombinant granulocyte-colony stimulating factor.

Five Thoroughbred and Quarter Horse cross foals were given 20 micrograms canine recombinant granulocyte-colony stimulating factor (rcG-CSF) per kg bwt intramuscularly (i.m.) on the day of birth and 10 micrograms rcG-CSF/kg for 13 additional days. During this time and for an additional 21 days haematology, bone marrow and clinical chemical analyses were performed. After one day of rcG-CSF administration leucocyte and neutrophil counts increased from 9.16 x 10(9)/l to 23.44 x 10(9)/l and from 6.45 x 10(9)/l to 19.61 x 10(9)/l, respectively. The counts continued to increase for the next 3-4 days and then there was a slight decrease. A second increase followed and the leucocyte and neutrophil counts increased to 52.84 x 10(9)/l and 45.16 x 10(9)/l on the day after the last rcG-CSF administration (Day 15). The counts decreased rapidly immediately after the administration of rcG-CSF was stopped and then at a slower rate. The cell counts were still higher than in the controls at the end of the study period (Day 35). Bone marrow cellularity increased from 10-25% before rcG-CSF was given to 60-80% after 5 days. The increase in cellularity was due to increased myeloid activity because the myeloid to erythroid ratio increased from 2.7 to 8.8. Serum chemistry changes were minimal although foals given rcG-CSF at various times had lower glucose concentrations and increased alkaline phosphatase and gamma glutamyl transferase activities.

Alkaline Phosphatase↗