Frequent expression of the cell death-inducing gene Bax in Reed- Sternberg cells of Hodgkin's disease

The expression of a cell death-inducing gene, Bax, was investigated in 52 cases of Hodgkin's disease in parallel with Epstein-Barr virus and was compared with the immunodetection of other apoptosis-regulating proteins, Mcl-1, Bcl-2, and Bcl-x. Bax immunostaining was found in 92% of the cases, among them 28% with a strong signal in more than 75% of the Reed-Sternberg cells. Mcl-1 was positive in 80% of the cases, whereas Bcl-2 and Bcl-x were found in 53% and 88% of the cases, respectively. Of 48 (89%) Bax-positive tumors, 43 were found to express apoptosis-inhibiting proteins such as Mcl-1 or Bcl-2. With the exception of 1 case, all Bax-positive tumors also expressed either Bcl- 2, Bcl-x, Mcl-1, or combinations of these anti-apoptotic proteins. No correlation was found between Bax expression and the presence of apoptotic cells as detected by morphology and the in situ 3' OH-DNA end- labeling technique. Our findings show that the apoptosis-inducing gene Bax expression is frequently expressed in Hodgkin's disease, providing a potential explanation for the good chemoresponses generally obtained for patients with this neoplastic disorder.     

Detection of factor XIIIa (active fibrin-stabilizing factor) in normal plasma

Factor XIIIa (active fibrin-stabilizing factor) generated in heat- defibrinated plasma by the addition of thrombin can be measured by 14C- putrescine incorporation into casein. Modification of this assay be substituting 3H-putrescine of high specific activity as the donor amine permits measurement of amine incorporation by plasma even in the absence of added thrombin. Incorporation is calcium dependent, inhibited by iodoacetamide, and absent from congenital factor XIII- deficient plasma and from normal platelets. The transamidating activity detected by radioenzymatic assay catalyzed the formation of gamma-gamma dimers and alpha polymers of fibrin and was thus biologically functional. This fibrin cross-linking activity was absent from factor XIII-deficient plasma. These experiments show (1) some factor XIII is present in plasma as factor XIIIa; (2) this factor XIIIa can cross-link fibrin and thus has biologic activity as well; and (3) this activity is not present in factor XIII-deficient plasma. Factor XIIIa in normal plasma is possibly activated in vivo, perhaps by circulating thrombin, factor Xa, or other proteolytic enzymes.     

Marrow- and spleen-seeding efficiencies of all murine hematopoietic stem cell subsets are decreased by preincubation with hematopoietic growth factors

The cobblestone-area forming cell (CAFC) assay permits a direct measurement of the seeding of primitive and more mature murine hematopoietic stem cell subsets by comparing the number of CAFC in the original transplant with the number of CAFC retrieved from bone marrow (BM) and spleen after transplantation. We found no differences in seeding efficiency between the more mature and primitive CAFC subsets, nor between seeding efficiencies of stem cells from low-density (LD) fractions of normal and day-6 post-5-fluorouracil BM. The data show that 18% to 20% of all intravenously transplanted stem cell subsets seed to the BM, whereas 8% to 10% seed to the spleen. In addition, similar seeding efficiencies were found for day-12 spleen colony- forming unit (CFU-S-12) as was determined by retransplantation. Previously, it has been reported that a 2- to 3-hour preincubation of BM with interleukin-3 (IL-3) enhances the in vivo repopulating ability of a graft. To test whether hematopoietic growth factors affected this increased engraftment by enhancing the seeding of the transplanted marrow, we assessed the 16- to 18-hour seeding efficiency of short- and long-term in vivo repopulating stem cell subsets to BM and spleen using the CAFC assay, after preincubation with or without hematopoietic growth factors. A 2- to 3-hour preincubation with IL-3, or a combination of IL-3, IL-12, and steel factor, at 37 degrees C, led to a substantial decrease in seeding compared with control (which was kept on ice) of all hematopoietic subsets measured, both in spleen and BM. In concert with these data, the long-term in vivo repopulating ability of growth-factor incubated BM was also decreased when compared with control. In conclusion, we have been unable to observe a beneficial effect of growth factor preincubation on the repopulating ability of a graft.     

Recombinant gibbon interleukin 3 supports formation of human multilineage colonies and blast cell colonies in culture: comparison with recombinant human granulocyte-macrophage colony-stimulating factor

The genetic sequences encoding the gibbon and human interleukin 3 (IL 3) proteins were molecularly cloned. The amino acid sequence of the mature gibbon IL 3 protein proved to share 93% homology with the corresponding human protein. We examined the effects of biosynthetic (recombinant) gibbon IL 3 on the proliferation and differentiation of an enriched population of human hematopoietic progenitors and compared the results with the effects of recombinant human granulocyte- macrophage colony-stimulating factor (GM-CSF). Gibbon IL 3 supported the formation of various types of single lineage as well as multilineage colonies by My-10+ bone marrow cells in the presence of human erythropoietin (Ep). In contrast, recombinant human GM-CSF supported the formation of single-lineage colonies and only a small number of multilineage colonies. Both IL 3 and GM-CSF had significant erythroid burst-promoting activity (BPA). Delayed addition of gibbon IL 3 to low serum culture of My-10+ marrow cells supported the formation of blast cell colonies with variable but high replating capability. Human GM-CSF was less effective than IL 3 in support of multipotential blast cell colonies. These results are analogous to the effects of murine IL 3 and GM-CSF on murine progenitors and support the notion that the primary factor for multipotential progenitors is IL 3.     

Diagnostic application of two-color flow cytometry in 161 cases of hairy cell leukemia

Recent immunophenotypic studies of hairy cell leukemia (HCL) have suggested specific patterns of immunoreactivity that may aid in diagnosis. We studied peripheral blood (PB) from 161 cases of HCL using two-color direct immunofluorescence flow cytometry and an extended panel of antibody combinations. Circulating hairy cells were identified by immunophenotypic features in 92% of the cases and could be detected even when representing < or = 1% of circulating lymphocytes. The 133 cases with > or = 2% detectable hairy cells were analyzed in detail. HCL showed a uniform and unique B-cell phenotype, with each of the following features identified in 99% to 100% of cases: (1) positive staining for B-ly7, coexpressed with CD19; (2) very intense, uniform expression of CD11c, with CD19; (3) moderately intense staining for CD25, with CD19; (4) very intense staining for CD22; (5) moderate to very intense staining for CD20; and (6) moderately intense monoclonal surface Ig. Phenotypic variability existed in expression of CD10 (26%) and CD5 (4%). Based on these features, HCL was easily distinguished from 50 cases of chronic lymphocytic leukemia (CLL). Although CLL exhibited frequent expression of CD11c (74%) and CD25 (68%), the intensity of staining was significantly less than HCL. Furthermore, CLL was uniformly positive for CD5 and showed weak staining for CD20, CD22, and surface Ig. B-ly7 proved to be the most specific marker, reacting with 100% of HCL cases, but absent in all cases of CLL. We conclude that two-color flow cytometry with specific antibody combinations is an efficacious method for characterization and sensitive detection of hairy cells in PB. Application of the phenotypic criteria described should help to increase accuracy in diagnosis of HCL.     

Expression of the alloantigen Zwa (or P1A1) on human vascular smooth muscle cells and foreskin fibroblasts: a study on normal individuals and a patient with Glanzmann's thrombasthenia

The cytoadhesin family consists of platelet glycoprotein (GP) IIb-IIIa and the endothelial vitronectin receptor. The beta subunit (GP IIIa) of these complexes expresses the alloantigen Zwa (or PIA1). This alloantigen is not expressed by members of other integrin subfamilies. By using immunoprecipitation and immunoblot techniques, we found that the beta subunit of a heterodimer, expressed by cultured human arterial smooth muscle cells and cultured foreskin fibroblasts, carries the Zwa antigenic determinant. Furthermore, the mobilities of the alpha and beta subunits of these two heterodimers are indistinguishable from those of the alpha and beta subunits of the endothelial vitronectin receptor. Therefore, we propose that the smooth muscle cell and fibroblast heterodimer are members of the cytoadhesin family. In Glanzmann's thrombasthenia, platelet GP IIb-IIIa is absent or severely reduced. Previously, we showed that endothelial cells from a thrombasthenic patient normally synthesize and express a GP IIb-IIIa- related molecule (the vitronectin receptor). Here we show that arterial smooth muscle cells, obtained from the same patient, express a surface molecule indistinguishable from the endothelial vitronectin receptor. We also demonstrate that both the endothelial and the smooth muscle cell GP IIIa-related molecule in this Glanzmann patient express Zwa. Our data indicate that (a) GP IIb-IIIa-related molecules on cell types other than platelets and endothelial cells can express Zwa in vitro, and (b) patients with Glanzmann's disease can express the Zwa antigen. This study substantiates our view that the defect in Glanzmann's disease is restricted to the megakaryocytes/platelets.     

Role of 18F‐FDG‐PET/CT in the management of marginal zone B cell lymphoma

The use of PET in patients with marginal zone B cell lymphoma (MZL) is controversial because of variability of fluorodeoxyglucose (FDG) avidity. We analyzed 40 PET/CT in 25 consecutive patients to compare its performance with CT at staging and as a first‐line response assessment. Sensitivity of PET/CT and CT was 96 and 76%. Mean standard uptake value was 6.1, 6.9 and 3.4 (p = 0.3) in nodal, extranodal and splenic subtypes, respectively. Of 17 patients (extranodal: n = 9; nodal: n = 6; splenic subtype: n = 2) with both imaging tests available at diagnosis, 8 (47%) had more involved areas with PET/CT than with CT, 75% of which were extranodal lesions. PET/CT resulted in upstaging of five patients although treatment of only two of them was changed. Responses of 15 patients with post‐treatment PET/CT were the following: 9 negative and 6 positive of which 3 were isolated residual lesions. Progression was documented in two of these three patients. Response was also assessed by CT in 11 patients. Discrepancies were found in three: Two were in complete remission by CT while PET/CT detected localized residual disease; another patient was in partial remission by CT, whereas PET/CT showed only one positive lesion. Two of these three patients relapsed. Patients with negative post‐treatment PET/CT did not relapse. With a median follow‐up of 50 months (10–152 months), 3‐year overall survival was 100 and 80% for patients with negative and positive post‐treatment PET/CT (p = 0.2). Three‐year disease‐free survival was 86%; the negative predictive value (NPV) was 100%, and the positive predictive value (PPV) was 83.3%. Although a larger number of patients will be required to further confirm these data, we can conclude that PET/CT is a useful imaging tool for both staging and response assessment in patients with nodal and extranodal MZL as a result of its high sensitivity, NPV and PPV. Copyright © 2014 John Wiley & Sons, Ltd.     

Randomized control trial of an integrated therapy for comorbid anger and gambling

Abstract

This study evaluated an integrated treatment for comorbid problem gambling, anger, and substance use. Problem gamblers with comorbid anger problems (N=42), half of whom also had substance use disorders, were randomized to either a 14-week integrated treatment targeting anger and addictions (i.e., both gambling and substance use) or a specialized treatment-as-usual (TAU) for gambling and substance use. Participants were assessed at baseline (Tl), 14 weeks (T2), and 12 weeks follow-up (T3). Relative to the TAU, participants in the integrated anger and addictions treatment reported significantly less gambling at T2 and T3 and less trait anger and substance use at T3. Findings suggest that it is important to screen gambling clients for the presence of comorbid anger and substance use problems and that, when present, these problems need to be addressed concurrently in gambling treatment in order to optimize treatment outcomes.