bluuebunny/crossref_metadata_2025 huggingface dataset:
Solitary Plasmacytoma. Presentation of 22 Cases of Two Centres from Buenos Aires, Argentina
Authors: Dragosky Marta , Alcaraz Susana , Anetta Isabel , Corzo Ariel , Devotto Laura , Deza Ernesto Gil , Luchetta Patricia , Marquez Marisa
Published: April 2020

Solitary plasmacytoma is a rare plasmatic cell tumor. Plasma cell neoplasia usually are generalized disease, in a small number of cases, less than 5 %, presents as a single bone lesion: solitary bone plasmacytoma (SBP) or develops outside the bone: extramedullary plasmacytoma (EMP).

Diagnostic criteria are: single bone tumor or single extramedullary mass of clonal plasma cells, normal bone marrow, no evidence of disseminated disease on skeletal survey, no anemia, hypercalcaemia or impairment of renal function, absent or low serum or urinary monoclonal immunoglobulin.

Material and Methods We present 22 patients treated at the Departament of Hematology of Marie Curie Hospital and Henry Moore Institute, 13 with SBP and 9 with EMP diagnosis. All were studied with the criteria of multiple myeloma. Prevalence of male 3.4/1 in both. Ages: SBP: 27–68 range, medium 47.23, EMP: 29–74 range, medium 55.

Locations of SBP: 5 in rachis (D3, D5, D7, D7, D8), 2 in clavícula, 2 in humerus, 1 in femur, sternum, etmoides, frontoparietal. The EMP: 7 in the head and neck: 3 in sinuses, 2 oropharynx, 1 uvula, maxillary mass (in area irradiated by NHL 7 years before), 1 lung + amyloid, 1 inguinal mass. Presence of a monoclonal paraprotein in serum/urine, was founded in 6 SBP (46%): 2 IgG kappa, 2 IgG lambda, 2 IgA lambda and in 2 EMP (22%): 1IgG lambda, 1 light chain lambda.

Results Treatment: SBP: in 10 RT + QT (VAD, Tal/Dex), 2 RT, 1 QT. Surgery was used to stabilize spine and/or long bones. EMP: 4 surgery + RT, 3 QT (VAD, Tal/Dex), 1 surgery + QT, 1 QT + RT. 8 patients underwent autologous bone marrow transplantation, 7 with SBP and 1 EMP, 5 of them aged under 36 years SBP: 3 patients achieved CR (10, 39, 77 months), 1 had local bone relapse, 2 without follow up, 3 of 7 transplanted patients relapsed, with evolution to multiple myeloma, 8 months, 23 months and 7 years later; the other 4 remain in CR, 27 to 82 months, media 36. EMP: 8 of 9 remain in CR, 2 to 120 months, media 35. One patient had a ileocecal relapse (initial site: oropharyngeal).

Conclusions Our series present male predominance, as reported by other studies. SBP prevail in axial skeleton and long bones in 61.5 %. Presents a worst development and prognosis, even though after ABMT. EMT was most frequently localized in head and neck (77,7%), in upper airways and oral cavity. The evolution was favorable, with only a single relapse. Diverse therapeutic methods happen to be useful. Low monoclonal paraprotein was present in 46 % of BPS and 22 % EMP.
A Clinical Study of Bortezomib in Combination with Dexamethasone and/or Doxorubibcin for the Treatment of Patients with Relapsed or Refractory Multiple Myeloma
Authors: Jin Jie , Qian Jiejing , Meng Haitao , Qian Wenbin , Tong Hongyan , Tong Yin , Mao Liping
Published: April 2020

Purpose: This study assessed the safety, tolerability, and response rate of bortezomib in combination with dexamethasone and/or doxorubibcin in patients with relapsed or refractory multiple myeloma.

Methods: Botezomib was administered 1.3mg/m2 on days 1, 4, 8, 11 of a 28-day cycle. Intravenous dexamethasone was administered 20–40mg/d on day 1–2, 4–5, 8–9,11–12, and 5 patients were also given 20mg/d doxorubibcin on day 1–4.

Results: Thirty relapsed or refractory myeloma patients (table 1) were enrolled; two patients could not be evaluated because they were lost to follow-up. Of the remaining patients 19 are male, 9 are female. Median age of the patients were 61(49–78). Median number of prior therapies was 4 (1–11). 22 of 28 patients who had been evaluated were assessable for response according to the EBMT criteria (European Group for Blood and Marrow Transplantation criteria). One CR(3.6%), twelve immunofixation-positive CRs[nCR] (42.9%), six PRs(21.4%), and three MR (10.7%) were observed. All patients who had an objective response were alive as of this analysis. Nine patients died during follow-up. Mean response time (time to best response)for the 28 patients was 25 days. Most common Grade ≥3 toxicities (table 2) were peripheral neuropathy(3/28), thrombocytopenia (3/28), rash(one in 28 patients). Six patients(21.4%) suffered herpes after one or two cycles.

Table 1. Patents and Disease Characteristics (N=28)

No. % Abbreviations: Ig, immunoglobulin; Parameter 61 49–78 Age, years Median Range 19 Sex Male Female 9 Paraprotein type IgG IgA Light-chain only 18 β2..Cmicroglubulin, 3 64.3 10.7 Range 7 25.0 Prior treatments 1262–8691 Median 4 Range 1–11

Table 2. Treatment-emergent adverse events

Total No. of patients with event(%) Adverse event 0 Hematologic Neutropenia No. with 3(10.7) grade ≥3 event Thrombocytopenia No. 0 with grade ≥3 event Anemia No. with 6(21.4) grade ≥3 event 5(17.8) Nonhematologic herpes Diarrhea 10(35.7) Fatigue Rash No. with 1(3.6) grade ≥3 event Tachycardia Peripheral 1(3.6) neuropathy Total No. No. 16(57.1) with grade ≥ 3 event 3(10.7)

Conclusion: Bortezomib plus dexamethasone given on a 28-day schedule showed encouraging activity with manageable toxicity and represents a promising treatment for multiple myeloma patients.
Toxicity of Bortezomib: Is There An Influence of Application Rates of Doxorubicin and Dexamethason Administration?a Clinical Monitoring
Authors: Steinborn Wolfram , Stein Wolfgang , Klebe Sandra , Kiehl Michael
Published: April 2020

Background: Bortezomib has shown significant anti-tumor activity in multiple myeloma. There are varied protocols including bortezomib, doxorubicin and dexamethason with different application rates. We observed in a retrospective analysis a wide range of toxicity, so that we have to modified our standard approach. The purpose of our analysis was to assess the frequency of toxicity and their alteration after changing the protocol.

Patients and methods: Eleven patients were treated for two 28-day cycles using our standard protocol. Bortezomib was given at 1.3 mg/m2 (days 1,4,8,11) and dexamethason at 40 mg (days 1–4, 9–12, 17–20) as well as doxorubicin at 9 mg/m2 (days 1–4). Seven patients received a third modified cycle composed of Bortezomib at 1.3 mg/m2 (days 1,4,8,11), dexamethason at 40 mg days 1–4 and doxorubicin at 20 mg/m2 days 1 and 4.

Results: One patient had unergone prior autologous transplantation, the other ten patients (90%) were newly diagnosed. All patients reported moderate peripheral neuropathic pain during the first two cycles. Eight patients (72%) developed severe adverse effects, twenty-five percent during the first cycle and seventy-five percent after the second one. The age of the affected patients range from 65 to 77 years (average age 70.5 years), the mean age of the unaffected is 64.7 years (54 to 72 years). Grade 3–4 toxic effects included infections (100%), neutropenia (25%), acute renal insufficiency (25%) and pulmonary edema (1 patient). Within the group of infections a pneumonia were diagnosed in six patients (75%), herpes zoster in two patients (25%) and an erysipelas in one patient. Two patients (25%) with pneumonia are died from a septic-toxic shock. The seven patients receiving the modiefied third cycle were without any grade 3–4 toxicity.

Conclusions: The age seems to have an influence on adverse effects (p=0.19) but there is a high significant benefit concerning the toxicity of bortezomib depending on application rates of doxorubicin and dexamethason.
No Evidence for Increased Prevalence of JAK2 V617F in Women with a History of Recurrent Miscarriage
Authors: Dahabreh Issa J. , Jones Amy V. , Voulgarelis Michael , Alafakis-Tzannatos Christina , Giannouli Stavroula , Varla-Leftherioti Marighoula , Keramitsoglou Theodora , Zoi Christine , Loukopoulos Dimitris , Fotiou Stelios , Cross Nicholas C.P. , Zoi Katerina
Published: April 2020

Myeloproliferative neoplasms (MPN) have been associated with increased rates of pregnancy complications. Women with essential thrombocythemia (ET), and, much less frequently, polycythemia vera (PV) present at a childbearing age and it has been reported that up to a third of MPN pregnancies may fail as first trimester miscarriages. A recent study has demonstrated that the JAK2 V617F mutation, seen in nearly all PV cases and half of those with ET, was significantly more common in women with a first unexplained pregnancy loss compared to controls, although the overall incidence was low (1.06% of women with a first unexplained pregnancy loss versus 0.2% in controls, p<0.001). We hypothesized that JAK2 V617F may be further enriched in women who had multiple miscarriages. We studied peripheral blood leukocyte DNA from 389 women who had suffered at least three consecutive spontaneous early miscarriages (<10 weeks gestation) or at least one late pregnancy loss (>10 weeks) by a tetra-primer amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) assay with a sensitivity of detection of 1% JAK2 V617F. 92% of women had a history of early miscarriage and 14.1% of late miscarriage. Detailed clinical information was available for 230 (59%) cases: the mean age was 35.9 years (standard deviation=5.4; range: 20–50) and the median number of miscarriages was 3 (range: 3–7). Factors known to be associated with recurrent miscarriages were excluded. The JAK2 V617F mutation was not detected in any case, yielding a 95% confidence interval for the prevalence of the mutation ranging from 0 to 0.009 and excluding a 1% prevalence of JAK2 V617F in this population (p=0.038 for the overall population; p=0.057 for early miscarriages only; 2-tailed Clopper-Pearson exact test). Our study therefore demonstrates no enrichment of JAK2 V617F among unselected women with a history of recurrent miscarriages. Although we cannot exclude the possibility of a small, significant excess of V617F over background levels, our findings indicate that a latent maternal JAK2 V617F-positive MPN is an unlikely cause of miscarriage.
Flow Cytometry Analysis of Peripheral Blood CD34+ Cells in Patients with Primary Myelofibrosis
Authors: Agarwal Archana M , Samuelson Scott James , Preobrazhensky Sergey , Parker Charles J. , Hickman Kimberly , Perkins Sherrie L. , Prchal Josef T. , Bahler David W.
Published: April 2020

Primary myelofibrosis (PMF) is a clonal chronic myeloproliferative disorder characterized by the accumulation of megakaryocytes in the bone marrow (BM), variable degrees of BM fibrosis, tear-drop erythrocytes, increased numbers of CD34+ hematopoietic progenitors in the peripheral blood (PB), and extramedullary hematopoiesis. Since the antigenic properties of the circulating CD34 cells may yield clues to disease pathogenesis and have not been extensively studied, we used five-color flow cytometry to examine these cells from 20 well characterized patients with PMF and 10 normal controls. Bone marrow biopsies, molecular and cytogenetic studies were also reviewed. As expected, the percentages of peripheral-blood CD34 cells were significantly higher in the PMF patients (mean 1.4%, range, range 0.065–7.15) compared to the controls (mean 0.05%, range 0.01–0.57). The mean fluorescence intensity (MFI) values related to HLA-DR expression were increased (more than 3 fold) on the CD34+ cells in 12/20 (60%) PMF patients relative to normal control levels, while increased levels of CD13 were seen in 5/20 (25%) of PMF patients. CD33 and CD117 expression were similar on the CD34+ cells in both groups. Aberrant expression of lymphoid antigens was observed in 6/20 (30%) with CD7, 6/20 (30%) with CD4, and 3/20 (15%) with CD56 on CD34 positive cells in PMF. In the18 cases also studied with antibodies against CD45RA and CD123, the majority of CD34+ CD38 + cells phenotypically resembled megakaryocyte-erythroid precursors (CD45RA−, CD123−) in 5 cases, common myeloid progenitors (CD45RA−, CD123+) in 12 cases, and granulocyte-macrophage progenitors (CD45RA+, CD123 +) in 1 case. JAK2-V617F mutations were detected in 9 of 20 cases, but were present in only 1 of 5 cases showing predominately megakaryocyte-erythroid precursors. The percentage of CD34+ cells also expressing CXCR4 (CD184) appears to be increased in some patients relative to normal controls in contrast to other reported studies. In conclusion, the peripheral blood CD34+, progenitor cells in PMF patients are heterogeneous phenotypically resembling megakaryocyte-erythroid precursors in approximately 30% of cases, and common myeloid progenitors in approximately 70% of cases. In addition, these cells often show phenotypic abnormalities (increased intensity of HLA-DR and CD13 expression) that can be detected with flow cytometry relative to normal peripheral blood CD34+ cells. Patterns of antigen expression in PMF also appear to differ from those reported for CD34 positive cells in other myeloproliferative disorders which may help in early diagnosis and/or monitoring treatment responses.
Enhancment of SCA1 Positive Hematopoietic Progenitors in Polycythemic Mice Inhibits Leukemogenesis
Authors: Ben-David Yaacov , Usenko Tatiana , Haeri Mehran , Vecchiarelli-Federico Laura , Prchal Josef T.
Published: April 2020

PV is a myeloproliferative disease, displaying a pronounced increase in the number of erythroid cells. Despite this increase, PV patients exhibit a lower incidence of acute leukemia and in particular erythroleukemia compared to its related disorder CML. In this study we induced erythroleukemia in a mouse model of familiar polycythemia, carrying a truncated form of the human erythropoietin-receptor (Epo-R). Previous studies have shown that these mice develop a disease similar to patients with familiar polycythemia, from whom this gain-of-function receptor was isolated and introduced as transgene. We show that the progression of erythroleukemia induced by Friend virus is significantly delayed in this genetic mouse model of polycythemia. These mice exhibit a massive enrichment of Sca1+/cKit− progenitors and mature immune cells compared to normal mice. Co-transplantation experiments revealed that Sca1+/cKit− progenitors inhibit the robust tumorigenic potential of Sca1−/cKit+ erythroleukemic cells.

In order to determine the mechanism of erythroleukemia inhibition, we established cell lines from Sca1+/cKit− cell populations and demonstrated a similar anti-leukemic inhibition in vivo. These cells also express cell surface markers for early B-cells (B220) and macrophages (Mac1), and are able to undergo differentiation into macrophages and dendritic cells following treatment with GM-CSF and IL4. We have shown that these cells also secrete factors that inhibit proliferation of erythroleukemic cells in culture, such as Nitric Oxide (NO). Accordingly, an NO inhibitor can suppress the inhibitory effect of the Sca-1+/cKit−/B220+/Mac1+ cells. To further demonstrate the importance of these leukemic inhibitory cells, we induced a polycythemia in normal mice through administration of Epo. Epo administration delayed the development of Friend virus-induced erythroleukemia and significantly increased the number of leukemic inhibitory Sca1+ cells. Our study also demonstrates a significant increase in the number of leukemic inhibitory cells with anti-proliferative effects in response to Epo and granulocyte macrophage-colony stimulating factor (GM-CSF) stimulation. We therefore propose that administration of Epo and GM-CSF in combination with NO agonistic drugs, as well as the identification of novel drugs that can induce the inhibitory Sca-1+/cKit−/B220+/Mac1+ cells, may lead to improved therapies for the treatment of myelodysplastic syndrome. Together, our results identify Sca-1+/cKit−/B220+/Mac1+ cells as potent cellular inhibitors of leukemia and unravel a potentially novel therapeutic approach.
Sequence Analysis of MPL Exon 10 of ET and MF JAK2 V617F Negative Patients Reveals a New Mutation
Authors: Bento Celeste , Duarte Marta , Almeida Helena , Menezes Cristina , Estevinho Alexandra , Coucelo Margarida , Kaeda Jaspal , Ribeiro Maria Leticia
Published: April 2020

Background. Diagnosis and classification of myeloproliferative disorders was greatly enhanced by reports describing JAK2V617F mutation in Polycythemia Rubra Vera (PRV) patients and approximately 50% of Essential Thrombocythemia (ET) and Myelofibrosis (MF) patients. More recently mutations mapping to the juxtamembrane region of the thrombopoietin receptor MPL were described in 5–10% of the patients with MF and 1% of the patients with ET. The reported data imply the Exon 10 MPL mutations lead to receptor activation in the absence of thrombopoietin binding and constitutive phosphorylation of JAK2, STAT3, STAT5, AKT and ERK. However, thus far only mutations mapping to codons 515 and 505 have been published. The aim of this study was to determine the frequency of JAK2V617F and MPL exon 10 mutations in MF and ET patients seen at the Hematology Department, Centro Hospitalar de Coimbra.

Methods. Sixty three patients with ET and 12 with MF, diagnosed according to the Polycythemia Vera Study Group (PVSG) criteria, were studied. Genomic DNA was extracted from peripheral blood samples collected into EDTA. Samples were tested for JAK2 V617F mutation by allele specific polymerase chain reaction (ASO-PCR). Subsequently these samples were screened for MPL exon 10 mutations by direct sequencing.

Results. 45 (71.4%) of the 63 patients with ET and 6 (50%) of the 12 MF patients had JAK2V617F mutation. Of the remaining 6 MF patients 3 had MPL mutations; two with W515L and one with W515K amino acid substitutions. Direct sequencing failed to detect exon 10 MPL mutations in 17 of the 18 ET JAK2 V617F negative patients. The remaining patient had a previously unreported MPL mutation mapping to codon 524 where C was substituted by T, predicting a replacement of arginine residue by cysteine (R524C).

Conclusions. JAK2 V617F mutation was detected in 71% of the ET patients and in 50% of the MF. MPL mutations were found in 2 out of 12 MF patients (17%) and in one of the 63 ET patients (1.6%). The MPL codon 515 mutations are reported to activate mpl protein independent of ligand binding. The prevalence of MPL mutations among MF patients is higher than the described in previous reports, however this maybe due to the small number of patients studied. Whilst there is genotype and phenotype correlation between MPD and MPL mutations described the precise functional role in the development of MPD remain unclear. Similarly, whether the previously unreported MPL R524C mutant allele presented here leads to activation of mpl protein like codon 515 mutations is unclear. But nonetheless such mutations assist in accurate diagnosis and classification of Philadelphia negative MPD patients with the view to optimization of clinical management of these patients.
JAK2V617F Mutation Status and Thrombosis in Essential Thrombocythemia
Authors: Cacciola Emma , Di Francesco Ernesto , Pezzella Francesca , Tibullo Daniele , Cacciola Rossella R
Published: April 2020

Thrombosis frequently complicates the essential thrombocythemia (ET) and hypercytosis may play a role in the its occurrence as well as the platelet and coagulant activation. Therefore, we evaluated myeloproliferative and platelet and coagulant activation markers such as platelets (plts), white blood cell (WBC), haemoglobin (Hb) and hematocrit (Hct) and β-thromboglobulin (βTG) and platelet factor 4(PF4) and prothrombin fragment 1+2 (F1+2), thrombin-antothrombin (TAT), tissue factor (TF), von Willebrand factor (vWF) and d-dimer (DD), respectively in ET patients and thrombosis. As has been reported that the Janus Kinase 2 (IAK2) V617F mutation gives ET a proliferative advantage, we also evaluated the JAK2 mutation. The study group consisted of fifty ET patients (24 males and 26 females, mean age 58 years) who fulfilled PVSG and WHO. Their mean duration of disease was 7 years. Of 50 patients, 18 were on hydroxyurea whereas 32 were not receiving any cytoreduction. All patients were on antiplatelets. Plts, WBC, HB and HCT were measured by automated analyser. βTG, PF4, F1+2, TAT, TF and DD and vWF were assayed by ELISA and immunoturbimetric assay, respectively. Jak2 mutation was analysed by melt curve analysis. Of 50 patients, 24 were JAK2 V617F-positive and 26 were wild-type (WT). Among JAK2-positive ET, 11 experienced thrombosis including 5 episodes of myocardial infarction (MI), 3 of deep vein thrombosis (DVT), 2 of transient ischemic attack (TIA) and 1 of erytromelalgia (E), whereas of WT ET, 5 developed thrombosis such as 1 DVT, 2 TIA and 2 E. All patients had thrombocytosis (684±299×109/l) and normal WBC (7.8±2.2×109/l) and HB(13±1.8 g/dl) whereas HCT and βTG were higher in JAK2-positive ET than WT (41±5.2 % vs 37±5.2 %) ( p=0.034) (266±83 IU/ml vs 193±18 IU/ml) (p<.0001). PF4 (113±39 IU/ml vs 6.9±2.2 IU/ml), F1+2 (3.4±3.6 nmol/l vs 0.7±0,2 nmol/l), TAT (16.9±28 μg/l vs 2.6±0.9 μg/l) and TF (93±132 pg/ml vs 6.4±2.8 pg/ml) (p<.0001 and p<.0001 and p=0.001 and p<.0001, respectively) were increased in all patients whereas vWF and DD were normal (56±42 % and 185±208 ng/l). There was no correlation between JAK2 mutation and plts and WBC and HB and any activation marker. A positive correlation there was between JAK2 mutation and HCT (p=0.034) and thrombosis (p=0.059). These data suggest that JAK2 V617F mutation is responsible for a hematopoietic expansion which might play a role in the pathogenesis of thrombosis in ET.
Concurrent Chromosomal Alterations at 3q27, 8q24 and 18q21 in An Atypical Form of Burkitt’s Lymphoma
Authors: Hamarshi Majdi SM , Kishk Maha abu , Mahafzah Mahmoud , Walloch Jami
Published: April 2020

Introduction: Chromosomal translocations are common in non-Hodgkin’s lymphomas (NHL), most frequently involving the genes bcl-2 in the t(14;18) of follicular lymphoma (FL), c-myc in the t(8;14) of Burkitt’s lymphoma (BL) and bcl-6 in the t(3;14) of follicular or diffuse large B-cell (DLBC) lymphoma. We report the clinical features, pathology and genetic findings in an exceedingly rare case of Burkitt’s lymphoma that showed concurrent involvement of these three chromosomal loci.

Case Report: This is a 65 year old Caucasian female who presented with a rapidly growing right supraclavicular lymph node over a few weeks. FNA biopsy showed typical morphology of Burkitt’s lymphoma. Similar morphologic features were found on the bone marrow biopsy. There was widespread disease with no CNS involvement. Flow cytometry from peripheral blood and immunohistochemistry on the cellblock showed B-cell phenotype positive for CD 10, CD19, CD20 (negative CD20 by immunohistochemistry), HLA-DR, cytoplasmic CD79a, and negative for CD34 and TdT. The interesting finding was the lack expression of surface or cytoplasmic immunoglobulin and expression of weak Bcl-6. Almost 90–95% expressed Ki67. The cytogenetic analysis reportedly demonstrated a complex karyotype t(3;8;14), and t(14;18) involving c-myc (8q24), bcl-2 (18q21), and bcl-6 (3q27). After 7 cycles of hyper CVAD-R she had bone marrow biopsy which showed residual disease. She also had a biopsy confirmed relapse as left arm nodule and left leg nodular infiltrate at 8 and 12 months form the diagnosis, respectively.

Discussion: This is a complex case of high grade B-cell lymphoma with morphology suggestive of Burkitt’s lymphoma. However the classification was challenging by the lack of surface immunoglobulin expression that might be expected in mature B-cell neoplasm “DLBCL, FL”, and the lack of TdT and CD34 that might be expected in precursor B-cell neoplasm “BL”. The diagnosis was highly dependent on the cytogenetic findings, which was significant for the presence of t(8;14) albeit in a three way translocation t(3;8;14), and t(14;18) involving c-myc (8q24), bcl-2 (18q21), and bcl-6 (3q27). The lymphoma was therefore described as “Burkitt’s transformation”. This is a rare translocation pattern, but has been described in follicular lymphoma, grade 3; diffuse large cell lymphoma; and Burkitt’s lymphoma.

Conclusion: BL might lack surface immunoglobulin expression making the diagnosis of high grade B-cell lymphoma challenging if based on the morphology and immunophenotyping alone. The cytogentetic findings better delineate sub-types of lymphoma. Molecular evidence of multiple oncogene deregulations, especially when involving the c-myc gene, appears to be associated with a dire clinical outcome.
Immunohistochemical Prognostic Marker for Diffuse Large B Cell Lymphoma in Patients Treated with CHOP Like Chemotherapy: Validation of the Algorithm by Hans Et Al. and Analysis of Individual Markers
Authors: Suh Cheolwon , Huh Jooryung , Oh Sukjoong , Jo Jeong-Hyeon , Kim Shin , Khang Joon
Published: April 2020

Background) Recently, many predictive models of the clinical outcome of diffuse large B cell lymphoma (DLBCL) have been proposed for better stratification of subgroups. However, studies of the protein expression have reported conflicting results, predominantly due to the lack of universally accepted cut-off value for positivity. Here we retrospectively analyzed the expression of CD10, BCL6, MUM1, BCL2, and CD5 in Korean DLBCL patients treated with CHOP like regimen using the widely used algorithm by Hans et al. (Blood 2004) and compared the results with different cut-off values. We also investigated the clinical significance of individual biomarkers.

Methods) Of the DLBCL cases retrieved from the file of Pathology, Asan medical center (1999–2004). Ninety four (94) cases with adequate tissue were included. Clinical characteristics, treatment and outcome of these patients were collected from the AMC data base. Immunohistochemical stains were performed on tissue microarray (TMA) sections of the archival material. According to Hans’ algorithm, cases were divided into germinal center B-cell like (GCB) (either CD10+ or BCL6+MUM1−)(19.1%) and activated B cell-like (ABC) (CD10−BCL6− or BCL6+MUM1+), with a universal cut-off value of 30% or more of the positively-stained tumor cells. The percentage of the positivity was also recorded in each case. The four cutpoints (0%, 1–30%, 31–50%, 51–75%, and 76–100%) were investigated for each marker for best correlation with the clinical outcome.

Results) During the 5-year median follow up, median overall survival was not reached. Hans’ algorithm divided the cases into 18 GCB (19.1%) and 76 ABC tumors (80.9%), with no clinical significance (3-year PFS: GCB group 63%, ABC group 62%). Among individual biomarkers, CD5− group (cut point: any positivity), bcl-6+ group (cut point: any positivity) and CD10+ group (cut point: 30%) had longer PFS, and bcl-6+ group (cut point: any positivity) had longer OS in univariate analysis. Multivariate Cox regression analysis showed CD10 (p=0.003) as independent prognostic marker in PFS; and bcl-6 (p=0.03) as independent prognostic marker in OS.

Conclusion) The present study shows the lack of predictability of Hans’ algorithm in DLBCL patients treated with CHOP like regimen among Koreans, while CD10, Bcl-6, and CD5 had the prognostic significance using different cut-off values. The result suggests that the proposed cut-off value may not be applied universally for best correlation, and that the optimal cut-off value may need to be optimized for individual laboratory.
Immunohistochemical Expression of Germinal Center and Non-Germinal Center Markers in Primary Gastric Diffuse Large B-Cell Lymphoma
Authors: Hurtado Fernando , Beltran Brady , Riva Luis , Salas Renzo , Morales Domingo , Carrasco Antonio , Quinones Pilar , Castillo Jorge
Published: April 2020

BACKGROUND: Recent studies have shown a correlation between diffuse large B-cell lymphoma (DLCBL) prognosis and molecular features using genome profiles by cDNA microarrays. Later reports have shown that immunohistochemical tests using markers as CD10, bcl-6 and MUM-1 to identify germinal center B-cell (GCB) and non-GCB patterns can have a similar value and are gaining major importance in assessing prognosis in patients with DLBCL. Gastric DLCBL is the most frequent extranodal non-Hodgkin lymphoma.

AIM: To evaluate the frequency of GCB and non-GCB subgroups in primary gastric DLCBL and the impact of these tests in prognosis.

PATIENTS AND METHODS: Patients older than 18 years with a diagnosis of primary gastric DLBCL were identified in a single institution from January 2002 to December 2005 and included in this analysis. Immunohistochemical stains were performed using antibodies against CD20, CD10, bcl-6 and MUM1 in all cases. Survival curves were obtained using the Kaplan-Meier method.

RESULTS: Twenty-nine patients were included in this study. The median age at diagnosis was 68 years (range 40 to 86 years). Thirteen male and 16 female patients were included. All cases received chemotherapy with CHOP regimen. Cases were subclassified in GCB and non-GCB using CD10, bcl-6, and MUM1 expression. Four cases (14%) were considered GCB and 25 cases (86%) non-GCB. There were no statistical differences between both groups in the following variables: age, ECOG performance status, serum LDH levels, clinical stage and International Prognostic Index (IPI) score. No statistical difference in survival was observed between the GCB and non-GCB groups. The 3-year overall survival for the entire group was 30%.

CONCLUSION: Immunohistochemical expression of non-GCB pattern is very frequent in primary gastric DLBCL in Peru. However, it does not show an effect in survival, probably related to the small number of cases included in the present study.
Age-Related EBV-Associated B-Cell Lymphoproliferative Disorders: Comparison with EBV-Positive Classical Hodgkin Lymphoma in Elderly Patients
Authors: Asano Naoko , Yamamoto Kazuhito , Tamaru Jun-Ichi , Oyama Takashi , Ishida Fumihiro , Ohshima Koichi , Yoshino Tadashi , Nakamura Naoya , Mori Shigeo , Yoshie Osamu , Shimoyama Yoshie , Morishima Yasuo , Kinoshita Tomohiro , Nakamura Shigeo
Published: April 2020

Age-related Epstein-Barr virus-associated (EBV+) B-cell lymphoproliferative disorder (LPD) is a disease group of EBV+ large B-cell lymphoma characteristically seen in elderly patients without predisposing immunodeficiency. This disease is now going to be listed in the up-coming WHO classification the 4th version. Age-related EBV+ LPD, especially of a polymorphous subtype, which occupied about one-third of cases, is often featured by a rich infiltration of reactive cells in the background, posing the differential diagnostic issue with classical Hodgkin lymphoma (cHL) associated with EBV. However, the clinicopathological differentiation between these two EBV-positive diseases remains to be elucidated. The aim of this study is to clarify the clinicopathological differences between the polymorphic subtype of age-related EBV+ LPD (n=34) and EBV+ cHL (n=108) of patients of 50 years and more of age. Age-related LPD was more closely associated with the following clinical parameters than cHL: a higher age at onset (71 vs. 63 years), lower male predominance (male:female ratio, 1.4 vs. 3.3), and a higher ratio of involvement of the skin (18% vs. 2%), the gastrointestinal tract (15% vs. 4%) and the lung (12% vs. 2%). Furthermore, age-related LPD was histopathologically characterized by a higher ratio of geographical necrosis, a greater increase (>30%) of cytotoxic T cells in the background lymphocytes, a higher positive rate for CD20 and EBNA2 together with an absence of CD15 expression than EBV+ cHL. As predicted by the clinical profile, age-related LPD had a significantly poorer prognosis than EBV+ cHL (P = 0.0001). This analysis has clearly demonstrated that the polymorphous subtype of age-related LPD constitutes an aggressive group with an immune response distinct from those of EBV+ cHL.
Bilateral Optic Nerve Invasion of Extranodal Marginal Zone B Cell Lymphoma with Leukemic Subtype
Authors: Sung Hwa Jung , Kim Eui Bae , Lee Se Ryeon , Park Kyong Hwa , Choi In Keun , Oh Sang Chul , Kim Byung Soo , Kim Yeul Hong , Kim Jun Suk , Seo Hee Yun , Kim Sun Hyung , Yoon Soo-Young
Published: April 2020

Extranodal marginal zone B cell lymphoma(MZBL), leukemic subtype is a rare B cell neoplasm characterized by peripheral lymphocytosis and bone marrow infiltration without nodal involvement. We report the case of a man who presented a simultaneous bilateral optic neuropathy as the initial clinical manifestation of extranodal MZBL. A 62-year-old man was referred to the Neuro-Ophthalmology Department because of a decrease in visual acuity, dyschromatopsia, visual field defects. Optic disc swelling was present in both eyes with peripapillary hemorrhages. Bilateral anterior ischemic optic neuropathy was diagnosed. Marked visual improvement occurred after 24 hours of intravenous methylprednisolone therapy, and vision completely recovered within a week. Eight weeks later, vision deteriorated again with bilateral optic disc swelling. A magnetic resonance image of the brain and orbit showed only enhancement of the optic nerve. Complete blood count(CBC) studies revealed hyperleukocytosis (13699/uL) with 72 % lymphocytes. There was no anemia and thrombocytopenia. Bone marrow(BM) study showed hypercellular with diffuse infiltration of small lymphocytic cells comprising 65.5% of all nucleasted cells. On immunophenotyping of BM aspirate, lymphocytic cells were CD2-CD3-CD5-CD10-CD23-CD33-CD34-CD117-CD138-cMPO-CD7+CD13+CD19+CD20+CD22+CD45+CD79+FMC7+kappa+lamda-. Cervicothoracic & abdominal CT scan showed no hepatosplenomegaly and no systemic lymphadenopathy. Depending on findings of BM and CT scan, extranodal marginal zone B cell lymphoma, leukemic subtype was diagnosed. Lumbar puncture revealed a white blood cell count of 790/uL with 90% lymphocyte. Flow cytometry was significant for a predominant population of CD3-CD4-CD8-CD20+CD13+ B-cell lymphocytes, diagnostic of cerebrospinal fluid involvement by lymphoma. So we could diagnosed the patient as optic nerve invasion of MZBL without optic nerve biopsy. The patient took several intrathecal chemotherapy with MHA(methotrexate, hydrocortisone, cytosine arabinoside)and 4th systemic treatment with R-CVP(rituximab, cyclophosphamide, prednosone). After all the visual acuity were improved and the infiltrative lesion of lymphoma on both optic nerve was found improved on the following MRI of orbit. The laboratory finding of CBC and CSF were completely normalized.
Nasal-Type NK/T Cell Lymphoma: Clinical Features, Immunophenotype and Treatment Outcome
Authors: Qian Shenxian , Shi Pengfen , Gao DaQian , Chen Kuang , Tan Junfeng , Liu LiRong , Xie Yaping
Published: April 2020

The clinical characteristics and prognosis remain unclear for nasal-type NK/T-cell lymphoma. The aim of this study is to determine the clinical features and outcome. 26 patients diagnosed as nasal-type NK/T cell lymphoma were included in the analysis. Immunophenotype was analyzed by immunohistochemical staining for LCA, CD79α, CD20, CD56, CD3, CD45RO, All the cases of nasal NK/T cell lymphoma were LCA, The positivity rates of CD45RO and CD56 were 89.8%, and 66%, respectively. Tumor cells didn’t express antigens of B and histiocyte, According to Ann Arbor system, 2, 7, 12, and 5 patients had stage I, II, III, and IV. 24 patients received combined chemotherapy(CHOP plus Ara-c) and radiotherapy, and 2 patients received chemotherapy alone. The disease is characterized by predominant young males, a propensity for nodal involvement, frequent stage II–III diseases, low frequency of elevated LDH, low-risk international prognostic index (IPI), high sensitivity to radiotherapy, and intermediate sensitivity to chemotherapy. The 3-year overall survival and progression-free survival for all patients were 88% and 71%, respectively The age, B symptoms, stage, and IPI were important prognostic factors. Combined chemotherapy(CHOP plus Ara-c) and radiotherapy tended to improve the survival for patients with stage and II or III diseases. The distant extranodal dissemination were the primary failure patterns. nasal-type NK/T-cell lymphoma appears to have distinct clinical characteristics and favorable outcomes.
Assessment of FDG-PET Imaging in T-Cell and NK-Cell Lymphomas
Authors: Chalapud-Revelo Juan , Sobrevilla-Calvo Pedro , Carlos Cadena-Eumaña , Silvia Rivas-Vera , Omar Lopez-Navarro , Juan Labardini
Published: April 2020

BACKGROUND: Malignant T-cell lymphomas are a heterogeneous group of disorders that frequently involve extranodal regions. Treatment and prognosis depend on accurate staging and histological evaluation. PET is widely used in staging and response evaluation of B-cell lymphomas, however, the use of FDG-PET have not been well studied for T-cell and natural killer cell lymphomas, and there is controversy if PET can detect accurately extranodal tumor activity.

PATIENTS AND METHODS: We retrospectively evaluated FDG-PET in 38 patients with active T-cells, and T/NK-cells lymphoma diagnosed according to the World Health Organization (WHO) classification, from May 2007 to May 2008. We also evaluated the Standard Uptake values (SUV) according the different subtypes and the accuracy of PET in detecting tumor activity in the skin and nasal region.

RESULTS: Histological subtypes included were extranodal NK/T-cell lymphoma nasal type (ENKL, n=12); Anaplastic T-cell lymphoma (ATCL, n=11); peripheral T-cell lymphoma (PTL, n=7); mycosis fungoides and Sezary syndrome (MF/SS, n=5), angioimmunoblastic (An=3). FDG-PET detected a lymphoma lesion in at least one site in 35 out of 38 patients (91%). The positivity rate was high in all histological subtypes ENKL 92%, ATCL 100%, PTL 86%, MF/SS 80%, A 100%. Among 13 patients who had cutaneous lesions only 7 had FDG-avid skin lesions (sensitivity =53.8% and specificity=100%) and among 11 patients with nasal region lesions, 91% had FDG-avid cutaneous lesions (sensibility = 91%, specificity=100%). The SUV Value in the different subtypes is shown in the table and there was not statistical differences between the groups (p= 0.067)

Lymphoma N Mean SD(+/−) Range An 3 4.1 1.7 2.7–6 MF/SS 5 4.2 3.0 1.9–8.6 ATCL 11 8.3 5.3 1.6–20.8 PTL 7 3.5 1.6 2.2–6.0 ENKL 12 7.2 4.1 2.2–13.8 Total 38 6.35 4.3 1.6–20.8

CONCLUSIONS: Most of the T and T/NK cell lymphomas were detected by FDGPET. All cases, except one, of ENKL nasal type were FDG-avid, but only half of cutaneous lesions were detected by PET. In the whole T-Cell lymphomas group the mean SUV was 6.35, ATCL had the highest SUV, and we did not find any statistically differences between the groups. We conclude that PET has a high sensitivity for detection of active ENNK nasal type lymphomas and this information should be the basis for designing further studies to assess its value in staging, tumor response and as a prognostic factor.
Stage-Specific Switches in Alternative Pre-mRNA Splicing during Late Erythropoiesis Are Conserved from Mouse to Human
Authors: Gee Sherry , Villalobos Jonathan , Yamamoto Miki , Clark Tyson A. , Kang Jeong-Ah , Wickrema Amittha , Chasis Joel Anne , Conboy john G.
Published: April 2020

Spatial and temporal regulation of alternative pre-mRNA splicing determines which exons are incorporated into mature mRNA, modulating mRNA coding capacity to ensure synthesis of appropriate protein isoforms throughout normal differentiation and development. During erythropoiesis, a stage-specific switch in pre-mRNA splicing activates incorporation of protein 4.1R exon 16, thereby increasing 4.1R affinity for spectrin and actin and mechanically strengthening red blood cell membranes. We are exploring the hypothesis that stage-specific changes in pre-mRNA splicing regulate expression of other critical genes during terminal erythropoiesis. Last year we described exon microarray and RT-PCR studies that revealed several novel pre-mRNA splicing switches in terminally differentiating human erythroid progenitors. These alternative splicing events involved well-annotated exons with consensus exon-intron boundaries, supporting a model in which these events represent a regulated alternative splicing program rather than a breakdown of splicing integrity in late erythropoiesis. Here we report additional evidence for this model by showing that several erythroid stage-specific switches in alternative pre-mRNA splicing are conserved between human and mouse. Primary mouse splenic erythroblasts from FVA-infected mice were cultured in vitro under differentiation conditions and used as the source of RNA for analysis of murine erythroid splicing events. From a total of seven internal cassette exons whose splicing was activated in late human erythroblasts, five exhibited an analogous splicing switch in murine erythroblasts. Comparative genomic analysis showed that these alternative exons are embedded in regions of unusually high sequence conservation among vertebrate species, suggesting that important regulatory signals are contained within the adjacent introns. Indeed, the flanking introns for several of these exons contain binding motifs for Fox2, an RNA binding protein and known splicing regulator for many tissue-specific splicing events. Further analysis of the conserved erythroid splicing events revealed the following:

three splicing switches occur in transcripts encoding RNA binding proteins (MBNL2, HNRPLL, and SNRP70), suggesting significant changes in the RNA processing machinery of late erythroblasts; and three of these alternative exons encode premature stop codons that could induce nonsense mediated decay (NMD) and contribute to down-regulation of these genes during terminal erythropoiesis.

Consistent with the latter hypothesis, inhibition of NMD in murine erythroblast cultures led to increased accumulation of mRNA isoforms containing the premature stop codons. Together these results suggest the existence of a highly regulated alternative splicing program that is critical for late erythroid differentiation.
Activation of TAO2 Kinase by Arsenic Trioxide in Leukemic Cells
Authors: McNeer Jennifer L. , Dolniak Blazej , Kroczynska Barbara , Sassano Antonella , Platanias Leonidas
Published: April 2020

Arsenic Trioxide (As2O3) has major efficacy in the treatment of acute promyelocytic leukemia (APL), but its use in other malignancies is limited by the need for high intracellular concentrations to induce apoptosis. Prior work in our laboratory has demonstrated that the p38 MAP kinase (MAPK) pathway is activated following treatment of cells with As2O3 and exhibits negative regulatory effects on As2O3-induced apoptosis and growth suppression. In the current study, we sought to identify upstream effector mechanisms by which the p38 pathway is activated by As2O3 in leukemic cells. We found that the MAPK kinase kinase TAO2 (thousand and one amino acid protein kinase 2) is phosphorylated on Ser181 after treatment of NB4, NB4.306, and U937 cells with arsenic. Such phosphorylation was rapid, occurring as early as after 5 minutes of As2O3 treatment. In addition, our data indicate that such phosphorylation occurs downstream of As2O3-induced redox reactions, as demonstrated by increased phosphorylation in cells pretreated with the oxidizing agent buthionine sulfoximine (BSO) and decreased phosphorylation following pretreatment with the reducing agent dithiothreitol (DTT). Arsenic treatment of the cells also resulted in activation of the kinase domain of TAO2, as evidenced in in vitro kinase assay studies using ATF2 as an exogenous substrate. siRNA-mediated TAO2 knockdown resulted in inhibition of As2O3-induced p38 phosphorylation, suggesting that this kinase acts as an upstream effector of the arsenic-activated p38 MAPK pathway. Moreover, in studies to determine the functional relevance of TAO2 in the induction of As2O3-dependent antileukemic responses we found that siRNA-mediated TAO2 knockdown enhanced the suppressive effects of As2O3 on KT1-derived leukemic progenitor (CFU-L) growth in clonogenic assays in methylcellulose. Altogether, our data demonstrate that TAO2 is activated during arsenic treatment of leukemic cells lines and acts as an upstream activator of the p38 MAPK pathway. Such activation appears to occur in a negative feedback regulatory manner to compensate for the suppressive effects of As2O3 on leukemic cell growth. Importantly, these findings raise the possibility that targeting TAO2 may provide a novel approach to enhance the generation of the antileukemic properties of As2O3.
Differential Glucose Transporter Expression during Perinatal and Postnatal Erythropoiesis
Authors: Montel-Hagen Amelie , Blanc Lionel , Boyer-Clavel Myriam , Jacquet Chantal , Vidal Michel , Sitbon Marc , Taylor Naomi
Published: April 2020

Glucose is a major source of energy for living organisms and its transport in vertebrates is a universally conserved property. Of all cell lineages, human erythrocytes express the highest level of the Glut1 glucose transporter with >200,000 molecules/cell. However, we recently reported that erythrocyte Glut1 expression is a specific trait of vitamin C-deficient mammalian species, comprising only higher primates, guinea pigs and fruit bats (Montel-Hagen et al., Cell, 2008). We now show that in all other tested mammals, including mice, rats, dogs and cows, Glut1 is in fact transiently expressed in erythrocytes during the neonatal period. This is in marked contrast with humans, where Glut1 is present at equivalently high levels on both neonatal and adult RBC. In mice, we found that Glut1 expression was not associated with primitive erythropoiesis but was highly expressed during definitive fetal erythropoiesis. Indeed, this transporter was present at significantly earlier stages of erythropoiesis in fetal spleen and liver than immediately following birth. It was therefore important to determine whether erythrocyte Glut1 expression in mice is specifically associated with fetal erythropoiesis or alternatively, is common to any physiological state where an extensive erythropoiesis is provoked. Induction of a hemolytic anemia in adult mice resulted in a massive erythropoiesis with significant increase in glucose uptake but notably, Glut1 was not detected. Rather, in these conditions as well as following birth, Glut4, an insulin-sensitive transporter previously thought to be responsible for glucose uptake in muscle and adipose tissue, was highly expressed. Following birth, the concomitant repression of Glut1 and induction of Glut4 was associated with a significantly augmented ratio of the Sp3 to Sp1 zinc-finger transcription factors. Thus, in contrast to humans, murine Glut1 is highly expressed during definitive erythropoiesis and is then downregulated at birth. Further erythroid development is characterized by the upregulation of a distinct glucose transporter, Glut4. Expression of distinct glucose transporters in nonhuman erythrocytes, regulated at the transcriptional level, therefore characterizes different states of erythroid development and differentiation.