Histological alterations in bone marrow in patients with late engraftment after autologous bone marrow transplantation

Bone marrow histology after bone marrow transplantation has rarely been studied. Here, we reviewed the pre- and post-transplant bone marrow biopsies (BMB) of 40 acute myelogenous leukemia (AML) patients autografted in our center, 28 with normal and 12 with delayed peripheral recovery. The two groups were comparable in terms of previous therapy, disease phase and the number of infused cells, and received the same conditioning regimen. In the former group, reduced bone marrow cellularity and mild reticulin abnormalities were usual histological findings; in the latter, five patients had the same pattern, but the other seven had an almost undetectable hematopoietic parenchyma and severe reticulin derangement. One of these seven patients died of reactivated hepatitis B virus infection; the others eventually achieved peripheral recovery, with none of them experiencing a relapse. Autografted AML patients are excellent subjects for histological investigations. They account for the majority of delayed engraftments, the contribution of extramedullary components to the timing of engraftment is minimal, and leukemia relapse cannot be ruled out. These results suggest that BMB is a useful investigation in the work-up of late engraftment. A high degree of reticulin derangement with an almost undetectable hematopoietic parenchyma appear to be the morphological hallmarks of late engraftment.     

Bone marrow histology and CD34 immunostaining in the prognostic evaluation of primary myelodysplastic syndromes

The prognostic impact of bone marrow biopsy (BMB) histology and CD34 immunoreactivity was compared with that of the more conventional parameters (the FAB diagnosis, peripheral blood values, percentage of BM blasts and some common prognostic scores) in 100 MDS patients. Statistical correlations among the cytological, haematological, histological and immunohistochemical parameters and their relationship with clinical outcome were searched for. At univariate analysis, FAB classification ( P  < 0.001), pattern of blastic infiltration at BMB ( P  < 0.005), presence of CD34⁺ aggregates ( P  < 0.0005), percentage of blasts in BM aspirate ( P  < 0.0001) and percentage of CD34 positivity ( P  < 0.0001) proved to be linked to leukaemic transformation and, except for FAB classification, retained a high degree of prognostic significance in terms of survival. Leukaemic transformation occurred in 16/18 patients simultaneously presenting 'large' blastic infiltrates at BMB and CD34⁺ aggregates ( P  < 0.00001); 9/17 evaluable patients died within 12 months of diagnosis ( P  < 0.001). Discriminant functions for leukaemic transformation and survival did not offer any advantage over univariate analysis in the prognostic work-up. The results indicate that the size of blastic aggregates and CD34 positivity allowed patients with a worse prognosis to be identified irrespective of their FAB subtype, but the prognostic impact is considerably greater when both parameters are simultaneously taken into account, as testified by the restricted and homogenous subgroup of patients with both 'large' and CD34-positive aggregates.     

Topical delivery of nerve growth factor for treatment of ocular and brain disorders

Neurotrophins are a family of proteins that support neuronal proliferation, survival, and differentiation in the central and peripheral nervous systems, and are regulators of neuronal plasticity. Nerve growth factor is one of the best-described neurotrophins and has advanced to clinical trials for treatment of ocular and brain diseases due to its trophic and regenerative properties. Prior trials over the past few decades have produced conflicting results, which have principally been ascribed to adverse effects of systemic nerve growth factor administration, together with poor penetrance of the blood-brain barrier that impairs drug delivery. Contrastingly, recent studies have revealed that topical ocular and intranasal nerve growth factor administration are safe and effective, suggesting that topical nerve growth factor delivery is a potential alternative to both systemic and invasive intracerebral delivery. The therapeutic effects of local nerve growth factor delivery have been extensively investigated for different ophthalmic diseases, including neurotrophic keratitis, glaucoma, retinitis pigmentosa, and dry eye disease. Further, promising pharmacologic effects were reported in an optic glioma model, which indicated that topically administered nerve growth factor diffused far beyond where it was topically applied. These findings support the therapeutic potential of delivering topical nerve growth factor preparations intranasally for acquired and degenerative brain disorders. Preliminary clinical findings in both traumatic and non-traumatic acquired brain injuries are encouraging, especially in pediatric patients, and clinical trials are ongoing. The present review will focus on the therapeutic effects of both ocular and intranasal nerve growth factor delivery for diseases of the brain and eye.     

Efficient retrovirus-mediated transduction of primitive human peripheral blood progenitor cells in stroma-free suspension culture.

Retroviral transduction of hematopoietic cells has resulted in unsatisfactory gene marking in clinical studies. Since cytokine-stimulated stem cells have engrafted poorly in animal models, we investigated phenotypic changes during culture of peripheral blood progenitor cells (PBPC). Human CD34(+) HLA-DR(low) cells, immunomagnetically separated from PBPC collections, were found to extrude rhodamine-123, which is characteristic for primitive hematopoietic cells. Cells were grown in suspension cultures supplemented with cytokines. While interleukin-3-containing factor combinations promoted cell proliferation they caused loss of rhodamine-123 extrusion and reduced the frequencies of cobblestone area-forming cells (CAFC). Several other cytokines failed to stimulate cell divisions, which are required for retroviral transduction. A combination including Flt-3 ligand (FL), interleukin-6 and stem cell factor (SCF) preserved an immature phenotype for 5 to 6 days and stimulated cell divisions, which was improved upon addition of leukemia inhibitory factor and interleukin-11. Furthermore, the CAFC frequency among cells treated with these cytokines was increased as compared with widely used cocktails containing interleukin-3, interleukin-6 and SCF. Rhodamine-123 appeared to be a particularly sensitive indicator for differentiation of PBPC. For analysis of gene transfer, amphotropic retroviruses conferring an MDR1 cDNA were added repeatedly for 6 days to cytokine-treated PBPC stroma-free cultures. Proviral cDNA was detected by polymerase chain reaction in 68% of cobblestone areas derived from CD34(+)HLA-DR(low) cells that had been exposed to Flt-3 ligand, interleukin-6 and SCF. In summary, conditions were identified that facilitate efficient transduction of early PBPC with amphotropic retroviruses while preserving a primitive phenotype for extended periods.     

A long-term follow-up analysis in adult acute myeloid leukemia patients after hematopoietic stem cell transplantation

AIMS AND BACKGROUND: Over the last 17 years, 119 adult acute myeloid leukemia patients have undergone hematopoietic stem cell transplantation at our Center. STUDY DESIGN: Seventy patients in first complete remission received hematopoietic stem cell transplantation (28 allogeneic and 42 autologous HSCT) as late intensification after conventional chemotherapy; 38 patients received allogeneic hematopoietic stem cell transplantation in a more advanced phase. A reference group was built up by collecting 40 acute myeloid leukemia patients who received high-dose cytosine arabinoside as late intensification and whose complete remission lasted more than 10 months. RESULTS: Results of the study led to conclude that an earlier timing of allogeneic hematopoietic stem cell transplantation can be recommended in order to treat patients who would otherwise suffer an early relapse. CONCLUSIONS: The outcome of autologous hematopoietic stem cell transplantation in patients not in first complete remission supports the possibility of achieving good quality second complete remissions and suggests that autografting may be a life-saving strategy in selected acute myeloid leukemia patients with advanced disease.     

Double incontinence in urogynecologic practice: a new insight

OBJECTIVES: This study was undertaken to evaluate the prevalence of anal incontinence in an urogynecologic setting and to investigate the relationship between lower urinary tract dysfunction and anal incontinence. STUDY DESIGN: The study included 504 women referred to our urogynecologic outpatient clinic who were prospectively investigated and asked specific questions on anal incontinence. Clinical and instrumental data were compared between women with urinary incontinence and with double incontinence, with further analysis for subgroups in the anal incontinent group of women (passive/urge). For continuous variables, the Wilcoxon rank sum test was used, and the Fisher exact test was applied to dicotomic variables. Logistic regression was used for categorical data. A level of P<.005 was considered significant. RESULTS: Of the investigated women, 20.2% were also anally incontinent. Women with double incontinence showed higher scores for urinary urgency (P=.010), which reached the established level of significance only in the subgroup with urge anal incontinence (P=.003). In this group, a higher prevalence of detrusor overactivity was observed. CONCLUSION: Anal incontinence is highly prevalent among women with lower urinary tract disorders. The existence of subgroups of patients having different kinds of anal and urinary disorders should be taken into consideration both for research purposes and for new treatment perspectives.     

Immunohistochemical evaluation of bone marrow biopsies in myelodysplastic syndromes.

Immunohistochemical studies were performed with monoclonal antibodies (MAbs) reactive on paraffin embedded bone marrow biopsies in 19 patients with myelodysplastic syndromes, 8 of them during r gamma-interferon treatment. CD15 MAbs stained mature myeloid cells predominantly located close to the bone marrow trabeculae. Anti-gpIIIa MAbs permitted precise identification of megakaryocytic cells including precursors and dysplastic megakaryocytes. Labelling with CD45 and CD68 MAbs, recognizing lymphocytes and macrophages respectively, was intense in patients in steady state, but progressively decreased during leukemic transformation. Increase in CD45+ and/or CD68+ cells was also observed in most bone marrow biopsies after 3 months of r gamma-interferon therapy.     

Spleen alterations in hairy cell leukemia: a scanning electron microscopic study

In Hairy Cell Leukemia (HCL) peripheral blood and bone marrow cells show under the scanning electron microscope (SEM) a characteristic surface with numerous ruffles and microvilli. The spleen of a patient affected by HCL was studied by SEM after fresh sectioning and routine preparation. Cells with the typical "hairy" surface were observed infiltrating the red pulp, altering the normal reticular meshwork and causing red blood cell distortion. In the sinuses, hairy cells adhered to the endothelial cells causing sinus dilatation and destruction. Aggregates of hairy cells delimiting pooled erythrocytes were also observed and may represent the "pseudosinuses" described in previous light and transmission electron microscopic studies. These preliminary findings may explain the condition of hypersplenism which characterizes HCL. In addition, SEM is proposed as a rapid and simple method to identify HCL spleen involvement.