Association between atl and non-hematopoietic neoplasms

A high incidence of multiple primary neoplasms has been observed in our patients with ATL in comparison to persons with other forms of hematologic malignancy who we have observed during the past 23 years (1963–1985). Five of 15 patients with ATL (33·3 per cent) have had at least one other associated neoplasm in comparison to only 44 of 1156 patients with other forms of hematological malignancy (3·8 per cent). The incidence figures for secondary neoplasms associated with the other hematologic malignancies were 4·3 per cent (16/370) for acute non-lymphocytic leukemia (ANLL), 2·2 per cent (2/90) for acute lymphocytic leukemia (ALL), 4·8 per cent (1/21) for acute unclassifiable leukemia, 2·2 per cent (5/225) for chronic myelogenous leukemia, 4·7 per cent (2/43) for chronic lymphocytic leukemia, 5·9 per cent (8/136) for malignant monoclonal gammopathy and 3·7 per cent (10/271) for malignant lymphoma. The incidence of multiple neoplasms in patients with ATL in comparison to those with other hematological malignancies was statistically significant (p<0·01 or p<0·001). The neoplasms associated with ATL have been adenocarcinoma of the thyroid or stomach, and squamous cell carcinoma of the larynx, lip or lung. We identified ATL-derived factor (ADF) in the cytoplasm of the secondary neoplasms of the ATL patients by means of indirect immunofluoroscopy and immunohistochemical techniques utilizing anti-ADF antibody. We also identified ras p21 products in these neoplasms by means of p21 ras monoclonal antibody studies. The possibility that HTLV-I was the cause of the secondary neoplasms thus was investigated. HTLV-I provirus genome was not found in all the six cases of non-ATL leukemic cells of the patients with anti-HTLV-I antibodies as determined by means of Southern blot analysis utilizing pX DNA probe. These findings suggest that there is some association between ATL cells and pre-malignant cells through ADF or other unknown factors in the activation of ras oncogenes. Subsequent suppression of host immune defence mechanisms in ATL patients permits evolution of the secondary neoplasms.     

Upregulation of CD200 is associated with regulatory T cell expansion and disease progression in multiple myeloma

Immune dysfunction is an important feature of multiple myeloma (MM) leading to infections, enhancement of tumour growth and resistance to chemotherapy. The overexpression of CD200, expansion of T regulatory (Treg) cell and increased levels of immune modulatory cytokines like IL10, IL6 and transforming growth factor beta (TGFβ) were suggested to have a role in this context. The aim of this study was to assess CD200 expression, Treg percentage by flow cytometry and immune modulatory cytokines (IL10, IL6, TGFβ) by enzyme‐linked immunosorbent assay in MM patients at diagnosis. This study included 50 MM patients at diagnosis and 20 healthy controls. The positive CD200 expression was detected in 72% of MM patients. Among the CD200 positive group, 4/13 patients (30.8%) were classified as stage I, 18/23 (78.3%) were in stage II and 14/14 (100%) were in stage III; according to International scoring system. Treg percentage was significantly higher in stage III, followed by stage II then stage I (p < 0.01). Serum IL6, IL10 and TGFβ were significantly higher in MM patients as compared with controls (p < 0.01, p < 0.01, p < 0.05, respectively). The increased expression of CD200 and Treg percentages was associated with increased severity biomarkers (serum LDH and β2 microglobulin). The degree of CD200 expression was significantly positively correlated to Treg percentage (r = 0.565, p < 0.01). Analysis of the CD200 negative patients had a better progression free survival (p = 0.032) and overall survival (p = 0.04) as compared with those positive for CD200 expression. These findings illustrate a clear correlation between myeloma cell CD200 expression level and the frequency of immunosuppressive Treg cells. In conclusion, increased expression of CD200, expansion of suppressive Treg cells and elevation of cytokines might have a role in MM progression in this cohort of patients. Copyright © 2015 John Wiley & Sons, Ltd.     

Interleukin-4 inhibits proliferation of human leukemic megakaryoblast cell line mhh 225

The effects of six recombinant human cytokines: erythropoietin, GM-CSF, G-CSF, interleukin-3, -4 and -6 on the proliferation and differentiation of a human multilineage myeloid leukemia cell line MHH 225, established from the bone marrow of an AML(M7) patient in our laboratory determined by changes in antigen expressions using monoclonal antibodies in APAAP technique were examined in liquid suspension culture. The MHH 225 cells have been growing exponentially without cytokines or conditioned media. About 90 per cent of MHH 225 cells are CD33⁺CD34⁺CD3^–CD7^–CD19^–CD20^–TdT^– with 57.6 per cent, 28.3 per cent and 7.8 per cent of them being CD41⁺, glycophorin A⁺ and CD15⁺, respectively. After five days of treatment with erythropoietin, GM-CSF, G-CSF or IL-6 no change was observed in MHH 225 cell antigens expression. IL-3 (100 U/ml) induced a moderate increase in only CD13 and alpha naphthyl esterase positive cells from 6.5 ± 1.9 per cent and 5.7 ± 2.4 per cent in control cultures to 21.6 ± 3.0 per cent and 19.1 ± 2.8 per cent, respectively. On the other hand, 100 U/ml IL-4 significantly increased the number of CD13, CD15 and alpha naphthyl esterase positive cells to 48.9 ± 5.0 per cent, 47.2 ± 3.6 per cent and 46.1 ± 3.0 per cent, p < 0.001, respectively. Also, 100 U/ml IL-4 decreased the number of CD41 positive cells from 57.6 ± 2.8 per cent to only 25.9 ± 3.6 per cent and did not change the number of CD33 or glycophorin A positive cells. The present results showed that out of the six myelopoietic growth factors tested, IL-4 was the only one to inhibit selectively the proliferation of CD33⁺CD41⁺ leukemic megakaryoblast cells suggesting that IL-4 may have a lineage regulatory effect in favour of a myeloblastic CD33⁺CD13⁺CD15⁺ at the expense of a megakaryoblastic CD33⁺CD41⁺ amplification in human leukemia cells and with apparently no effect on leukemic erythroblast cells. The MHH 225 cell line provides a useful tool and freely available model to scientists for studying signal transduction via IL-4 and for studies of‘lineage switch’.     

ALL-TRANS RETINOIC ACID (ATRA) IN THE TREATMENT OF ACUTE PROMYELOCYTIC LEUKEMIA (APL)

Acute promyelocytic leukemia is characterized by the reciprocal translocation of chromosomes 15 and 17. All-trans retinoic acid (ATRA) efficiently induces differentiation of the abnormal promyelocytes. In this study, we had used ATRA as the primary induction therapy for 17 newly diagnosed patients, and as the salvage therapy for 11 patients who relapsed from or were resistant to chemotherapy. All patients received subsequent consolidation chemotherapy. Complete remission (CR) rate, early death rate (within 28 days of diagnosis) were then compared to an historical control of 50 APL patients treated with combination chemotherapy; and event-free survival of the 17 newly diagnosed patients was compared to the historical control. In the ATRA group, 26 of the 28 patients (93 per cent) attained complete remission. Two of 28 (7 per cent) died within 28 days of ATRA therapy. There was no case of primary resistance to ATRA. Combination chemotherapy was added to ATRA in five patients due to rapidly increasing leucocyte count. There was one case of retinoic acid syndrome which resolved with steroid. When compared to the 50 cases of historical control, there is significant improvement in the overall CR rate (92 per cent versus 59 per cent, p=0·001) and a significant reduction in the early mortality rate (7 per cent versus 41 per cent, p=0·001). Moreover, when the survival result of the 17 newly diagnosed patients were compared with the control, there is a significant improvement in the projected EFS at 3 years (64 per cent versus 25 per cent, p=0·007). In conclusion, ATRA was showm to improve the CR rate, reduce induction mortality and significantly prolong the event-free survival.     

Re‐evaluation of prognostic markers including staging,serum free light chains or their ratio and serum lactate dehydrogenase in multiple myeloma patients receiving novel agents

International Staging System (ISS), serum free light chain ratio (sFLCR) and lactate dehydrogenase (LDH) are well known, easily assessed independent prognostic indicators of outcome in multiple myeloma (MM). The purpose of the study was to re‐examine the prognostic contribution of these variables in a multicenter setting with special attention to MM patients treated with autologous stem cell transplantation (ASCT) or novel agents (NA). Three hundred and five symptomatic newly diagnosed MM patients were retrospectively studied. Twenty‐seven per cent, 32% and 41% were in ISS stages 1, 2, and 3, respectively. Fifty‐six per cent of them presented kappa light chain monoclonality; median sFLCR was 27.04 (0.37–1.9 × 10⁵) and 47.97 (0.26–2.3 × 10⁷) for kappa patients and lambda patients, respectively; patients with sFLCR above median constituted the high sFLCR group. Thirty‐one per cent of patients had increased LDH. As first line treatment, 55.7% received conventional treatment and 44.3% NA. After induction, 24% underwent ASCT, whereas 76% received NA at any line, either bortezomib (82.5%), thalidomide (48%) or lenalidomide (27%). When the 305 patients were analyzed together, staging, high sFLCR and abnormal LDH were predictive of survival. The same was true for patients that never received NA, whereas neither high sFLCR nor abnormal LDH constituted adverse factors in patients that received NA frontline. In the last group of patients, no difference was observed between ISS stages 2 and 3. The median 5‐year survival of patients that never received NA versus those who did frontline was 29% vs 47%, 7% vs 52% and 24% vs 40% in patients with abnormal LDH, high sFLCR and ISS stage 3, respectively (p = 0.03, p < 0.00001 and p = 0.035). In conclusion, patients gaining the most from NA are those with an aggressive disease as reflected by advanced stage, abnormal LDH and high sFLCR. In addition, the adverse impact of these three variables is obscured by NA. Copyright © 2012 John Wiley & Sons, Ltd.     

Serum concentrations and clinical significance of soluble CD40 ligand in patients with multiple myeloma

Multiple myeloma (MM) is a disease of plasma cells that express the CD40 receptor. Binding of the CD40 by its natural ligand, CD40 ligand (CD40L), produces growth arrest and/or apoptosis in MM. To evaluate serum levels of soluble CD40L (sCD40L) in MM patients and to correlate them with markers of disease activity and angiogenesis, such as vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), interleukin-6 (IL-6), proliferation marker Ki-67 proliferation index (Ki-67 PI) and bone marrow plasma cell infiltration, fifty-eight MM patients were studied in diagnosis and 43 of them after completion of treatment. Serum levels of sCD40L, VEGF, HGF and IL-6 were measured by ELISA, whereas Ki-67 PI and bone marrow plasma cell infiltration were measured by immunohistochemistry. Pre-treatment levels of sCD40L in MM patients were higher compared to controls and to their levels after effective treatment. Treatment regimen did not affect the degree of reduction of sCD40L levels, whereas patient in partial remission had increased levels compared to those with better response. Significant differences were found among disease stages. There were also positive correlations between CD40L with HGF, VEGF, IL-6 and Ki-67 PI. Elevated serum sCD40L is found in patients with advanced MM stage and can be reduced after effective treatment. Increased levels of this mediator are correlated with angiogenic cytokines, providing evidences that CD40L/CD40 interactions play a significant role in the mechanisms of angiogenesis in MM patients.     

Emerging roles of endoglin/CD105 and angiogenic cytokines for disease development and progression in multiple myeloma patients

Angiogenesis is an essential process for the expansion of multiple myeloma (MM), in which many angiogenic factors participate. Endoglin (CD105) is a transforming growth factor‐β co‐receptor, being mainly expressed in angiogenic endothelial cells and has been used as a marker of tumor angiogenesis, having prognostic potential. The aim of the study was to evaluate serum levels of soluble CD105 (sCD105) in MM patients, both during diagnosis and after effective conventional chemotherapy, in the plateau phase, and to correlate them with the clinical stage of the disease, as well as with the known angiogenic factors vascular endothelial growth factor, angiogenin and interleukin‐18 (IL‐18). Serum levels of the aforementioned factors were measured, by enzyme‐linked immunosorbent assay, in 56 newly diagnosed MM patients, in 35 of them who entered plateau phase and in 24 healthy controls. Bone marrow aspirations were also performed in all patients to determine plasma cell infiltration. All measured cytokines were higher in MM patients compared with controls and with advancing disease stage (p < 0.001 for all cases). Furthermore, the values of all factors decreased significantly in the plateau phase (p < 0.001 for all cases). Serum levels of sCD105 correlated with the other angiogenic cytokines, whereas only serum levels of angiogenin had prognostic value for the survival. In conclusion, CD105 and the angiogenic cytokines vascular endothelial growth factor, angiogenin and IL‐18, seem to have emerging roles both in angiogenesis and tumor growth in MM. Copyright © 2013 John Wiley & Sons, Ltd.     

Prognostic Biomarker Study in Pathologically Staged N1 Non-Small Cell Lung Cancer

Purpose: The prognostic influence of 6 biomarkers correlated to histologic subtypes of non-small cell lung cancer (NSCLC) on loco-regional control, overall survival, disease-free survival (DFS), and distant disease control (DDC) rates, all measured at 5 years, were examined.Materials & Methods: Cell blocks from the primary tumors of 137 patients with pathologically staged N1 NSCLC at MDACC were analyzed by 6-biomarker status correlated to histological subtypes and their outcomes.Results: The ranges of biomarker values were as follows: apoptotic index, 0.2–2.8%; mitotic index, 0–1.8%; the proportion of cells in S+G2M, 3–36%; p53 status, 0–100%; Ki-67, 0–9.3%; DNA index, 1.0–2.74. Subtypes of 137 cases from the postoperative pathology specimen showed that 74 patients had squamous carcinoma and 63 patients had adenocarcinoma. Mean and median lengths of follow-up were 4.21 years and 2.43 years, respectively. Patients with squamous cell carcinoma (SCC) had a better 5-year survival (p = 0.006), DFS (pp = 0.002) than patients with adenocarcinoma (AC). Among patients with AC, the DNA index was a significant predictor of 5-year DFS (p = 0.02), DDC rate (p = 0.04), and local-regional control (p < 0.05). Higher apoptosis (p = 0.03) and mitosis indices (p = 0.03) were also univariate predictors of increased distant disease among patients with AC. Multivariate analysis of patients with AC revealed that the DNA index and Ki-67 were the only significant independent predictors of distant metastasis (p < 0.04 and p < 0.02, respectively) and DFS (p < 0.04 for both). Among patients with SCC, univariate analysis showed that S+G2M proportion (p < 0.05) and Ki-67 levels (p < 0.02) were significant predictors for local-regional control; for SC, multivariate analysis showed that only mitosis was a significant predictor in this case for overall survival (p < 0.04).Conclusion: Spontaneous apoptotic index and Ki-67 were significantly higher in SC than in AC. Patients with SC had less distant metastasis better DFS and overall survival than those with AC. Multivariate analysis revealed that DNA index and Ki-67 status were significant predictors for DDC and DFS in patients with AC, but only mitotic index was a significant predictor of overall survival for patients with SCC.     

The use of immunohistochemistry to distinguish reactive mesothelial cells from malignant mesothelioma in cytologic effusions

BACKGROUND:

The distinction of benign from malignant mesothelial proliferations in cytologic specimens can be problematic. In this study, the authors investigated the utility of immunohistochemical (IHC) markers in making this distinction.

METHODS:

Archival paraffin‐embedded cell blocks of pleural and peritoneal fluids from 52 patients with malignant meothelioma (MM) and 64 patients with reactive mesothelial hyperplasia (MH) were retrieved. IHC stains included desmin, epithelial membrane antigen (EMA), glucose‐transport protein 1 (GLUT‐1), Ki67, and p53.

RESULTS:

Desmin was positive in 84% (54 of 64) cases of reactive MH and in 6% (3 of 52) of MM cases (P < .001). EMA was positive in 9% (6 of 64) of benign and 100% (52 of 52) of malignant cases (P < .001). GLUT‐1 was positive in 12% (5 of 43) of benign and 47% (7 of 15) of malignant cases. Ki67 showed strong nuclear positivity in >40% of mesothelial cells in 9% (6 of 64) of benign and 16% (8 of 49) of malignant cases (P = .38). p53 showed strong nuclear positivity in 2% (1 of 46) of benign and 47% (7 of 15) of malignant cases (P < .001). EMA positivity and desmin negativity were found in 2% (1 of 64) of reactive MH cases and 98% (49 of 52) of MM cases (P < .001). EMA negativity and desmin positivity were found in 86% (55 of 64) of reactive MH cases and 0% of MM cases.

CONCLUSIONS:

The combination of positive EMA and negative desmin strongly favors MM; conversely, a combination of negative EMA and positive desmin favors a reactive process. Likewise, strong membranous positivity for GLUT‐1 and/or strong nuclear staining for p53 favors a mesothelioma. Ki67 proliferative index showed no significant difference between reactive MH and MM cases. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Appearance of monoclonal plasma cell diseases in whole‐body magnetic resonance imaging and correlation with parameters of disease activity

The aim of our study was to assess in which way different infiltration patterns of monoclonal plasma cell diseases in whole‐body (wb) magnetic resonance imaging (MRI) are associated with clinical stages, plasma cell content in bone marrow samples and established serum markers of disease activity. Institutional review board approval was obtained. We performed wb‐MRI in 547 consecutive, unselected and untreated patients with monoclonal gammopathy of undetermined significance (MGUS, n = 138), smoldering myeloma (SMM, n = 157) and multiple myeloma (MM, n = 252) on two 1.5 T MRI‐scanners with body array coils. The studies were evaluated in consensus by two experienced radiologists blinded to the diagnosis. We observed focal lesions in 23.9% (MGUS), 34.4% (SMM) and 81.3% (MM), respectively. A diffuse infiltration pattern was detected in 38.4%, 45.9% and 71%, respectively. The differences between all infiltration patterns were significant (p < 0.0001). The presence of focal lesions and the presence of a diffuse bone marrow infiltration was associated with an increased plasma cell percentage in bone marrow samples (median 22% vs. 14%, 26% vs. 10%, both p < 0.0001) and monoclonal protein concentration (median 18 g/dl vs. 13 g/dl, p = 0.003, 20 g/dl vs. 11 g/dl, p < 0.0001). Further categorization of the diffuse infiltration patterns in wb‐MRI into “salt‐and‐pepper,” moderate and severe identified significant associations with M‐protein (median g/dl for S+P/moderate/severe 23/18/25, p = 0.04), plasma cell percentage in the bone marrow (median 25%/24%/40%, p = 0.02), and age (median years 67/60/57, p < 0.0001). Bone marrow infiltration in wb‐MRI is significantly different between the various stages of plasma cell disease and correlates well with established markers of disease activity.     

Expression of L‐type amino acid transporter 1 (LAT1) as a prognostic and therapeutic indicator in multiple myeloma

L‐type amino‐acid transporter 1 (LAT1) plays a key role in cell growth and survival. To determine the prognostic significance of LAT1 in multiple myeloma (MM), we investigated the expression of LAT1 and its functional subunit, 4Fc heavy chain (CD98), on myeloma cells by immunohistochemistry in 100 newly diagnosed MM patients. High expression (moderate or strong staining intensity) of LAT1 and CD98 was detected in 56% and 45% of patients, respectively. The LAT1 expression score was positively correlated with Ki‐67 index (r = 0.631, P < 0.001), and there was a statistically significant difference in Durie–Salmon stage between patients with high and low LAT1 expression (P = 0.03). In 43 patients treated with melphalan and prednisolone, the overall response rate was significantly higher in the high LAT1 expression group (60.0%) than in the low LAT1 expression group (17.6%) (P = 0.03). Multivariate analysis confirmed that high expression of LAT1 was a significant prognostic factor for predicting poor overall survival independently from the International Staging System (both P = 0.01). Here, we show that the overexpression of LAT1 is significantly associated with high proliferation and poor prognosis in newly diagnosed MM patients. Thus, LAT1 may be a promising pathological marker for identifying high‐risk MM.     

The prognostic significance of X-ray changes at presentation and reassessment in patients with multiple myeloma

The relationship between presenting skeletal X-ray findings and survival in multiple myeloma was assessed in 172 consecutive patients treated at this institute. All patients were investigated, treated and followed up according to one protocol. The shortest survival was seen in those with normal X-rays (3-year actuarial 11 per cent) and the longest in those with minimal lytic changes (3-year actuarial 44 per cent). Patients with osteoporosis alone and those with extensive lytic lesions had intermediate survivals of 32 per cent and 33 per cent respectively. Of the other major prognostic features only anaemia demonstrated a significant correlation with X-ray findings in terms of survival (p < 0.001) suggesting that those with normal X-rays may have a more diffuse marrow involvement accounting for their poor prognosis. The response on bone X-rays in 102 patients who completed nine cycles of chemotherapy was also analysed. Patients with evidence of healing had a 3-year actuarial survival of 37 per cent, those with progression 36 per cent and those with stable X-rays 65 per cent (p < 0.01). Normal bone X-rays at presentation and radiological evidence of healing following therapy appear to be bad prognostic signs in multiple myeloma.     

Epirubicin and docetaxel as first-line treatment for hormonal receptor positive metastatic breast cancer: the predictive value of luminal subtype: A retrospective cohort analysis

Abstract

Background: We retrospectively evaluated the efficacy of first-line epirubicin and docetaxel in patients with metastatic, hormonal receptor (HR)-positive, and human epidermal growth factor receptor-2-negative breast cancer. A subgroup analysis evaluated the predictive value of immunohistochemistry-defined luminal subtype.

Methods: We included patients with at least one visceral and measurable site of metastatic disease. Patients were grouped as luminal A (HR⁺ and Ki67<13%) or luminal B (HR⁺ and Ki67>13%).

Results: Forty-four patients were entered and prognostic variables were similar between the subgroups. Luminal B patients achieved higher objective response rate than luminal A (69% versus 19%; P?=?0·001), longer time to progression (12·2 months versus 8·6 months; P?=?0·039), and longer overall survival (24·6 months versus 19·5 months; P?=?0·041). The multivariate analysis confirmed the predictive value of luminal B subtype for longer time to progression.

Conclusions: Identification by Ki67 labelling index of human epidermal growth factor receptor-2-negative luminal A could predict a substantial benefit from systemic chemotherapy. Endocrine therapy would be the most appropriate therapy for luminal A tumours.     

Low frequency of apoptosis in Epstein-Barr virus-associated gastric carcinoma with lymphoid stroma

In order to understand the character of Epstein-Barr virus (EBV)-associated gastric carcinoma with lymphoid stroma (GCLS), we examined cell proliferation and death in comparison with gastric carcinoma without prominent lymphoid stroma (conventional gastric carcinoma). The EBV-encoded small RNA I (EBER-I) and apoptotic cells were examined by in situ hybridization (ISH) and by terminal deoxynucleotidyl tramferare (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL), respedvely, on formalin-fmed paraffin-embedded surgical speci-mens from 22 GCLSs and 23 conventional gastric carcinomas. lmmunostaining was performed for the detection of Ki-67 antigen (Ki-67), P53 protein (P53) and bcl-2 oncoprotein (BCL-2). The percentages of apoptotic and Ki-67-positive cells were expressed as apoptotic indices (Al) and Ki-67 labeling indices (KI), respectively. EBER- I was detected in I9 (86%) GCLSs in contrast to none in conventional carcinomas. Mean Al was 1.8 ± 0.6 in the EBER- I -positive GCLSs and 3.3 ± 1.7 in the conventional gastric carcinomas, the value being significantly lower in the former (p < 0.01). Mean KI was 40.0 ± 10.8 in the GCLSs and 48.5 ± 9.1 in the conventional gastric carcinomas, the value being significantly lower in the former (p < 0.05). P53 expression showed no significant difference between the 2 categories of carcinoma.     

Reinforced HEAV'D therapy for adult acute lymphoblastic leukemia: Improved results and revised prognostic criteria

Thirty-six adults with acute lymphoblastic leukemia (ALL) were treated with adriamycin, vincristine. prednisolone, and asparaginase for remission induction, followed by vincristine-adriamycin-cyclophosphamide consolidation courses, cranial irradiation, a short ara-C plus VM-26 pulse, and vincristine plus cyclophosphamide rotating weekly with ara-C plus VM-26 for three months (reinforced HEAV'D). Thirty-one patients achieved a complete remission (86 per cent). Compared with historical results from a prior study, age >30 years, absolute blast count >15 × 10⁹/1, and CD10-negative immunophenotype were not associated with higher relapse rate and shorter survival, suggesting a positive effect from intensification therapy with ara-C and VM-26 in these poor prognostic categories. However, patients with an abnormal karyotypic pattern or a positive molecular study for BCR-ABL rearrangement detecting t(9;22) had a far greater likelihood of treatment failure (probability of remission at 3 years 0·10) than those with normal karyotype or negative molecular study (probability 0·70), and those not studied or with insufficient methaphases (probability 0·50) (p<0·05 by log-rank test). These results underline the prognostic importance of chromosomal abnormalities and the usefulness of ara-C and VM-26 in the management of adult ALL.     

Effects of raloxifene on normal breast tissue from premenopausal women

Summary The objective of this study was to evaluate the effects of raloxifene on normal breast tissue. A randomized, double-blind study was carried out in 30 ovulatory, premenopausal women of 18–40 years of age, who had been diagnosed with fibroadenoma of the breast. The patients were divided into two groups: Group A (placebo, n=16) and Group B (raloxifene 60 mg, n=14). The medication was given for 22 days, beginning on the first day of the menstrual cycle. An excisional biopsy was carried out on the 23rd day during which a sample of normal breast tissue was collected to evaluate the presence of the proliferating cell marker Ki-67. Student’s t-test was used for the statistical analysis of data (p<0.05). Mean percentage of stained nuclei in groups A and B was 10.96 ± 1.27 and 1.21 ± 0.26, respectively (p<0.001). Raloxifene significantly reduced the proliferative activity of normal breast tissue in premenopausal women.     

Cellular Proliferation by Multiplex Immunohistochemistry Identifies High-Risk Multiple Myeloma in Newly Diagnosed,Treatment-Naive Patients

IntroductionTherapeutic options for multiple myeloma (MM) are growing, yet clinical outcomes remain heterogeneous. Cytogenetic analysis and disease staging are mainstays of risk stratification, but data suggest a complex interplay between numerous abnormalities. Myeloma cell proliferation is a metric shown to predict outcomes, but available methods are not feasible in clinical practice.Patients and MethodsMultiplex immunohistochemistry (mIHC), using multiple immunostains simultaneously, is universally available for clinical use. We tested mIHC as a method to calculate a plasma cell proliferation index (PCPI). By mIHC, marrow trephine core biopsy samples were costained for CD138, a plasma cell–specific marker, and Ki-67. Myeloma cells (CD138⁺) were counted as proliferating if coexpressing Ki-67. Retrospective analysis was performed on 151 newly diagnosed, treatment-naive patients divided into 2 groups on the basis of myeloma cell proliferation: low (PCPI ≤ 5%, n = 87), and high (PCPI > 5%, n = 64).ResultsMedian overall survival (OS) was not reached versus 78.9 months (P = .0434) for the low versus high PCPI groups. Multivariate analysis showed that only high-risk cytogenetics (hazard ratio [HR] = 2.02; P = .023), International Staging System (ISS) stage > I (HR = 2.30; P = .014), and PCPI > 5% (HR = 1.70; P = .041) had independent effects on OS. Twenty-three (36%) of the 64 patients with low-risk disease (ISS stage 1, without high-risk cytogenetics) were uniquely reidentified as high risk by PCPI.ConclusionPCPI is a practical method that predicts OS in newly diagnosed myeloma and facilitates broader use of MM cell proliferation for risk stratification.     

A comparative study of Ki-67 antigen expression between luminal A and triple-negative subtypes of breast cancer

Tumor biomarkers such as hormone receptors, HER-2 and Ki-67 are used routinely in clinical practice for classification of molecular subtypes of breast cancer. Cell proliferation evaluated by Ki-67 antigen expression is important to determine tumor aggressiveness. However, there is a paucity of studies comparing Ki-67 expression in an expressive number of cells among molecular subtypes of breast cancer, particularly among less and more aggressive tumors, such as luminal A and triple-negative, which have led us to the present study. The current study included invasive ductal carcinoma samples of 59 patients, which were divided into two groups: luminal A (n = 29) and triple-negative (n = 30). For immunohistochemical reaction, the samples were incubated with monoclonal anti-Ki-67 antibody (clone MIB1) and cells expressing Ki-67 protein were identified by dark brown staining of the nuclei, counting at least 600 cells per slide. The mean percentages of stained nuclei were analyzed by Student’s t test (p < 0.05). The mean percentage of nuclei stained with anti-ki-67 was 10.14 and 77.22 in luminal A and triple-negative breast cancers, respectively (p < 0.0001). Our study showed a high cell proliferation of triple-negative breast cancer in comparison with luminal A, justifying its aggressiveness and poor clinical outcome.     

Successful bone marrow transplantation in sensitized aplastic anemia patients using total lymphoid irradiation for conditioning: long-term follow-up

Between June 1986 and November 1994, 22 previously transfused patients with severe aplastic anemia (SAA) were treated with high-dose cyclophosphamide (CY) (50 mg/kg over 4 consecutive days) and 7 Gy total lymphoid irradiation (TLI) in two fractions before allogeneic bone marrow transplantation (BMT) from HLA-identical sibling. Graft-versus-host-disease (GVHD) prophylaxis included the combination of methotrexate and cyclosporine A in all cases. Actuarial survival at 5 years is 73±9 per cent for the entire group and 86±13 per cent for the seven patients ⩽18 years. The incidence of graft failure was 0 per cent, and of acute GVHD and chronic GVHD was 31·5 per cent and 24 per cent respectively. Prolonged interval from diagnosis to BMT adversely influenced survival (P=0·03). No hypothyroidism or secondary malignancies have been documented in this series. Our findings indicate that survival with CY-TLI is comparable to that obtained using preparative regimens without radiation. The changing role of radiotherapy in pretransplant immunosuppression for SAA is discussed. © 1996 John Wiley & Sons, Ltd.