Coenzyme Q10 and adriamycin toxicity in mice

Pretreatment for four days with coenzyme Q10 (CoQ10) significantly lowered the acute toxicity in female C3H/HeNCrlBR mice given moderately lethal (15.0 and 20.0 mg/kg) i.p. doses of adriamycin as well as in male ICR/Hla mice given 12.5 mg/kg i.p. adriamycin. In both strains of mice, CoQ10 pretreatment did not protect the mice at higher i.p. adriamycin dose levels. When adriamycin was administered by the clinically-used i.v. route, CoQ10 pretreatment did not reduce acute toxicity at moderately lethal doses in either strain. At higher i.v. adriamycin dose levels, CoQ10 pretreatment significantly enhanced acute toxicity. CoQ10 pretreatment did not alter the antitumor effectiveness of adriamycin (i.p. or i.v.) against the Dunn osteosarcoma.     

Reduction by coenzyme Q10 of the acute toxicity of adriamycin in mice

Pretreatment for four days with coenzyme Q10 (COQ10) reduced the acute toxicity in mice treated with adriamycin. In two sequential protocols, adriamycin allowed only 36 and 42% survival, respectively. Pretreatment with COQ10 allowed 80 and 86% survival, respectively. The differences are significant, p less than 0.05. The mechanism for this reduction in the acute toxicity may be based upon the prevention by the supplementary COQ10 of the inhibition caused by adriamycin to COQ10-dependent enzymes in cardiac and and other tissues. The prospect of diminishing the toxicity of adriamycin in cancer patients remains promising and important.     

Hematopoietic stem cells found in lineage-positive subsets in the bone marrow of 5-fluorouracil-treated mice

It is known that treatment of mice with 5-fluorouracil (5-FU, 150 mg/kg) confers radioprotection. To investigate this effect, we performed bone marrow transplantation (BMT) using C57BL/6-Ly5 congenic mice treated with 5-FU five days prior to experiments. The mononuclear cells (MNC) in 5-FU-treated bone marrow (BM) were 10 times more radioprotective than those in untreated BM. Moreover, the number of BM MNC expressing c-kit on their surface from 5-FU-treated mice was markedly decreased relative to those from untreated controls. These results showed that the surface characteristics of cells that contributed to this radio-protective effect differ from those of stem cells as reported recently. BM MNC of mice treated with 5-FU were separated on the basis of expression of the lineage-specific antigens (Lin), c-kit, and Ly6A/E. When injected into lethally irradiated mice, 1,000 Lin+ and Lin-c-kit+Ly6A/E+ cells showed radioprotective effects such that 100% and 60% survived, respectively. Flow cytometric analysis 165 days after BMT showed that 88.8% and 65.1% of peripheral blood (PB) in mice transplanted with Lin+ and Lin-c-kit+Ly6A/E+ was derived from donor mice, respectively. After six months, donor-derived Lin-c-kit+Ly6A/E+ cells which showed radioprotective effects on a secondary irradiated host were detected from mice transplanted with Lin+ cells from 5-FU-treated mice. Taken together, these findings demonstrated that stem cells expressing Lin+ present in the BM of mice treated with 5-FU other than Lin-c-kit+Ly6A/E+ cells and these Lin+ cells play an important role in the recovery of myeloablative mice.     

Immunological responses critical to the therapeutic effects of adriamycin plus interleukin 2 in C57BL/6 mice bearing syngeneic EL4 lymphoma

A safe and effective therapeutic combination of moderate doses of Adriamycin (doxorubicin, 4 mg/kg, IV, Days 1 and 8 or only Day 8) plus prolonged administration of moderate doses of interleukin 2 (2 micrograms, b.i.d., Days 9-40) was developed in the syngeneic EL4 (5 x 10(4) cells, IP, Day 0) lymphoma--C57B1/6 mouse model and has been reported in the companion paper. The studies described herein demonstrate that the effectiveness of this combination treatment against EL4 lymphoma growing intraperitoneally in C57B1/6 mice was dependent upon the presence of CD8+ cells. Thus, the induction of long-term survivors (60-80%) by Adriamycin plus interleukin 2 was completely ablated by pretreatment of mice with anti-CD8 monoclonal antibody (MAb), whereas pretreatment with anti-CD4 MAb only partially inhibited the therapeutic effects and anti-NK1.1 MAb had no effect. A close association between survival, an increase in phenotypically identified CD8+ cells, and an increase in specific anti-EL4 cytolytic activity was demonstrated with cells from the tumor site (peritoneum) but not consistently with cells from the spleen. No association was observed between survival and modulations in natural killer (NK), lymphokine-activated killer (LAK), or tumoricidal macrophage activity of spleen or peritoneal cells. Taken together the results indicate that, in this model, the most relevant correlate of a therapeutically effective host antitumor response is the level of specific EL4 tumor killing by cells present at the tumor site. Based on the findings reported herein, it can be predicted that weakly immunogenic tumors may be eradicated by immunologic mechanisms elicited in conjunction with properly designed therapeutic regimens.     

Growth and differentiation of bone marrow hematopoietic cells in mice bearing Ehrlich ascite tumor and treated with Dicyclopentadienildichlorotitanium (IV)

In this work we have investigated the growth and differentiation of bone marrow stem cells in mice bearing Ehrlich ascites tumor and treated with three dose-regimens of Dicyclopentadienyldichlorotitanium (IV) (DDCT). We also studied the presence of colony stimulating factors in the serum of DDCT-treated animals, as well as the effects of the drug on the survival of the tumor-bearing mice. The results demonstrated that the myelosuppression developed in the tumor-bearing animals is prevented by the administration of 1, 2 or 3 doses of 15 mg/kg DDCT. In the treatment with three doses, however, 23% of the animals died. Moreover, DDCT treatment in normal animals resulted in increased numbers of CFU-GM. We observed the presence of stimulating factors in the serum of drug-treated animals which induced the growth and differentiation of bone marrow progenitor cells from normal animals in vitro. On the other hand, in vitro addition of the drug to these cultures had no effect. Thus, we conclude that the drug protects against the myelosuppression induced by the tumor and that this protection may be related to an indirect action of the drug.     

Interleukin-5 expression in the bone marrow of sensitized Balb/c mice after allergen challenge

Interleukin-5 (IL-5) is a potent eosinophilopoietic factor implicated in the chronic inflammatory cell accumulation accompanying bronchial asthma. However, its role in stimulating eosinophil differentiation within the bone marrow following allergen exposure remains to be elucidated. The aims of our study were to determine the expression of IL-5 within the bone marrow of sensitized and control mice after allergen exposure, and to investigate the cellular phenotype of IL-5-producing cells. Sensitized Balb/c mice were challenged with either ovalbumin (OVA) or sterile saline. After 6 h, the mice were exsanguinated and the bone marrow prepared for cytospins. Bone marrow-derived cells from OVA-sensitized mice exhibited an increase in IL-5 immunoreactivity and mRNA compared with those from nonsensitized control mice (p < 0. 05). After allergen challenge, there was a further increase in IL-5 expression (p < 0.05) within the bone marrow. Both sensitization and allergen challenge resulted in an increase in the number of cells expressing major basic protein (MBP) (p < 0.05). In nonsensitized mice, the IL-5 mRNA was expressed predominantly by CD34-positive (CD34+) progenitor cells. Following sensitization and allergen challenge, CD3-positive (CD3+) T lymphocytes were the major source of this cytokine. These results demonstrate the presence of IL-5 within the bone marrow of normal Balb/c mice. After sensitization and allergen challenge, the increase in IL-5-producing cells within the bone marrow is attributed by T lymphocytes.     

Preclinical toxicity of liposome-incorporated annamycin: selective bone marrow toxicity with lack of cardiotoxicity

Annamycin (Ann) is a new lipophilic anthracycline antibiotic with a marked ability to circumvent typical multidrug resistance both in vitro and in vivo. Because of its high affinity for lipid membranes and very low solubility in water, Ann has been prepared in a submicron liposome formulation (L-Ann) that is currently being investigated in a Phase I clinical study. We studied the preclinical toxicity of L-Ann in mice and beagle dogs and compared it with that of free Ann in suspension and the parent compound doxorubicin (Dox). In mice, free Ann was about twice as toxic as Dox (LD50 after a single i.v. bolus administration, 8.8 versus 19.9 mg/kg; P < 0.01). The liposomal carrier reduced Ann toxicity by 2-fold (LD50, 15.74 mg/kg for L-Ann versus 8.8 mg/kg for free Ann; P < 0.01). Granulocytopenia was the main toxicity of Ann, either free or liposome incorporated, and was much more profound than with an equitoxic dose of Dox as assessed by blood counts and pathological studies. In chronic mouse studies, L-Ann was remarkably less cardiotoxic than Dox. Cumulative toxicity with the weekly administration of a given fraction of the subacute LD10 was markedly higher with Dox than with L-Ann as assessed by body weight and mortality studies. L-Ann also had less vesicant toxicity than Dox after intradermal administration in mice. Beagle dogs tolerated the mouse-equivalent LD10 dose of L-Ann (1.4 mg/kg) with no side effects, changes in the hematological and biochemical blood parameters, or pathological changes. Our results indicate that: (a) L-Ann is more selectively myelotoxic than Dox and is noncardiotoxic; (b) the liposome carrier plays a major role in the favorable toxicity profile of L-Ann; and (c) the standard one-tenth of the LD10 should be a safe starting dose for Phase I clinical trials with L-Ann in humans.     

Clozapine treatment of HIV-associated psychosis--too much bone marrow toxicity?

Benign cystic mesothelioma of the peritoneum is uncommon and usually occurs in women. We report this condition in a man, who was treated successfully by surgical excision of the tumor.     

Hypercalcemia as the first manifestation of bone marrow T-cell lymphoma

The authors describe the case of a 20-year-old patient where the first leading symptom was hypercalcaemia. A similar case was not published so far in the Czech literature. The disease took a fulminant course and proved fatal nine days after the first symptoms of the disease. The correct diagnosis was established only by necropsy. The adverse course of the disease could not be influenced by repeated haemodialysis nor by the administration of disodium pamidronate (Aredia) and calcitonin. The authors discuss differential diagnostic problems of hypercalcaemias and the pathogenesis of hypercalcaemia in malignant diseases of the haematopoietic system.     

系统性间变性大细胞淋巴瘤骨髓累及的临床病理学研究

目的 探讨系统性间变性大细胞淋巴瘤(S-ALCL)骨髓累及的临床病理学特点、免疫学表型及临床生物学行为.方法 回顾性分析34例S-ALCL病例资料,进行骨髓活检(19例)或涂片(15例).其中ALK(+)24例,ALK(-)10例.HE染色、免疫组织化学染色观察病理形态及免疫表型,原位杂交法检测EB病毒.结果 6例(17.6%)S-ALCL存在骨髓累及,均经骨髓活检标本确定,15例患者骨髓涂片中均未见肿瘤累及.ALK(+)ALCL和ALK(-)ALCL骨髓累及的发生率分别为16.7%(4/24)和20.0%(2/10),差异无统计学意义(P=0.3555).与无骨髓累及病例比较,骨髓累及病例的年龄、性别分布差异无统计学意义(P值分别为0.8089和0.3085).骨髓累及者肿瘤细胞以间质性分布为主[83.3%(5/6)].生存分析统计提示伴有骨髓累及的患者预后明显差于无骨髓累及者(P=0.0407).结论 S-ALCL骨髓累及发生率低,与患者的发病年龄、性别及ALK蛋白的表达无相关性.伴有骨髓累及的S-ALCL患者临床预后差,骨髓活检在判断S-ALCL预后中有重要意义.     

Morphologic, immunophenotypic, and molecular evaluation of bone marrow involvement in non-Hodgkin''s lymphoma

AIM: To determine the tumour proliferative activity in a series of archival cerebral astrocytomas and compare proliferating cell nuclear antigen (PCNA) and Ki-67 labelling indices in the primary and recurring neoplasms following therapeutic radiation. METHOD: Twenty eight cases of pre-irradiated and post-irradiated astrocytomas (ranging from WHO grades I to IV) were stained immunohistochemically using the avidin-biotin horseradish peroxidase technique. Two antibodies, PC10 and MIB-1, were used to establish the proliferating labelling indices, PC10 identifies PCNA and MIB-1 recognises the Ki-67 antigen. RESULTS: Both antibodies showed significantly higher labelling indices in the post-irradiated specimens. However, in general, the Ki-67 indices were lower than those for PCNA. MIB-1 immunoreactivity showed less variation and was more intense than that seen with PC10. The discrepancy between the labelling indices of the pre-irradiated and post-irradiated samples raises questions about the evolution of astrocytomas and the effects of therapeutic intervention. CONCLUSIONS: The data may represent genetic alterations, the natural tumour course, and/or the effect of radiation. Although both of the antibodies reflected the state of growth of neoplastic cells in astrocytomas, MIB-1 was more reliable. A simple immunohistochemical method using proliferation markers does have an important role in the future care of patients with astrocytoma.     

Cells capable of bone production engraft from whole bone marrow transplants in nonablated mice

Allogeneic and autologous marrow transplants are routinely used to correct a wide variety of diseases. In addition, autologous marrow transplants potentially provide opportune means of delivering genes in transfected, engrafting stem cells. However, relatively little is known about the mechanisms of engraftment in transplant recipients, especially in the nonablated setting and with regard to cells not of hemopoietic origin. In particular, this includes stromal cells and progenitors of the osteoblastic lineage. We have demonstrated for the first time that a whole bone marrow transplant contains cells that engraft and become competent osteoblasts capable of producing bone matrix. This was done at the individual cell level in situ, with significant numbers of donor cells being detected by fluorescence in situ hybridization in whole femoral sections. Engrafted cells were functionally active as osteoblasts producing bone before being encapsulated within the bone lacunae and terminally differentiating into osteocytes. Transplanted cells were also detected as flattened bone lining cells on the periosteal bone surface.     

Allogeneic bone marrow transplantation for relapsed and refractory Hodgkin''s disease and non-Hodgkin''s lymphoma

There is national and international interest in increasing the community-based component of undergraduate medical education, but more research is needed on its potential, practicability and effectiveness. The objective of the study was to examine the feasibility and efficacy of general practitioners teaching basic clinical skills to first year clinical medical students in the community. The structure and methods of evaluation of the programme are described. Evaluation tools included semistructured interviews of general practitioner tutors; student questionnaires; assessment of student performance; and costs of the programme. The great majority of the students found the programme enjoyable (81 out of 81, 100%) and educational (79 out of 81, 97%). Students' performance in the end of rotation Objective Structured Clinical Examination suggested that clinical skills are acquired at least as well in the community as in the hospital. Tutors identified the personal benefits of this teaching as development of their own clinical skills and the stimulation of teaching. The programme has been successfully expanded from 24 students to 230 students annually and has demonstrated that community-based teaching can usefully contribute to undergraduate medical education in the area of clinical skills teaching. Key practical issues for schools contemplating similar initiatives are presented.     

Indium bone marrow scintigraphy as an aid in selecting marrow biopsy sites for the evaluation of marrow elements in patients with lymphoma

One hundred and two previously treated lymphoma patients were studied with 111Indium bone marrow scans and bone marrow biopsies. The biopsies were considered to represent sampling errors when the cellularity of the biopsy did not reflect the general state of the marrow organ cellularity as demonstrated by the scan. In each instance the accuracy of the scan was confirmed by either another biopsy or the subsequent clinical course of the patient. Sampling errors were infrequent (1/51) in patients with normal peripheral blood counts and whose marrow had never been involved with tumor. Errors were especially likely (17/51) in patients who had had marrow involvement or those who had anemia, leukopenia, or thrombocytopenia. The 111Indium bone marrow scan allows the clinician to avoid selecting a biopsy site with a high risk for sampling error.     

Bone marrow cell development and trabecular bone dynamics after ovariectomy in ddy mice

This review highlights recent progress in our understanding of the beneficial effects of hormone replacement therapy (HRT) in cardiovascular disease (CVD). The fact that HRT is increasingly advocated has raised concern about possible adverse effects weighed against the potential benefits of HRT regimens. Both favourable and unfavourable effects of oestrogens and HRT regimens on CVD risk factors are increasingly recognized. Consequently, the picture on cardiovascular effects of oestrogen and HRT has become more complicated, and research in this field has extended to novel areas.     

Immunohistochemistry in apparently normal bone marrow trephine specimens from patients with nodal follicular lymphoma

AIM: To establish the role of immunohistochemistry (using a limited panel of antibodies) in detecting minimal involvement by follicular lymphoma in routinely processed bone marrow trephine specimens, which show no obvious morphological (light microscopic) evidence of lymphoma; to determine whether bcl-2 immunostaining in bone marrow distinguishes between benign and malignant infiltrates in a patient with nodal follicular lymphoma. METHODS: Twenty seven consecutively selected paraffin wax embedded, formalin fixed bone marrow trephine specimens were stained with the following antibodies: anti-bcl-2, anti-CD79a, anti-CD3, and kappa and lambda light chains, using the Streptavidin biotin complex technique. RESULTS: Five of the 27 cases, which showed no evidence of involvement by follicular lymphoma on routine stains, showed monotypic B cells on immunohistochemistry. Two of the cases were diffuse, while the remaining three showed mini-aggregates around bony trabeculae. In all five cases the lymphomatous infiltrates were strongly bcl-2 positive. Reactive B lymphoid nodules did not show the same degree of bcl-2 positivity, and negative cells could be discerned within the reactive nodules. CONCLUSIONS: There is merit in studying so-called negative bone marrows immunohistochemically in order to detect minimal involvement by follicular lymphoma. A limited panel of antibodies including anti-bcl-2, anti-CD79a and anti-CD3 is usually adequate to accomplish this. Strongly bcl-2 positive lymphoid aggregates in the bone marrow of patients with nodal follicular lymphoma are indicative of lymphoma.