bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Mesenchymal Stem Cells Express HLA-G: Role in MSC Mediated Immunosuppressive Effect.
Authors: Chapel Alain , Nasef Aisha , Mathieu Noelle , Frick Johanna , Bouchet Sandrine , Gorin Norbert C. , Thierry Dominique , Fouillard Loic 
Published: October 2019

Abstract
Mesenchymal stem cells (MSC) possess immunomodulatory properties, being able to suppress allogenic T cell response and modify maturation of antigen presenting cells. Recently its role for the treatment of graft versus host disease has been reported. The mechanisms of immunosuppression induced by MSC are being currently investigated.
HLA-G is a non-classical major histocompatibility complex (MHC) class I antigen HLA-G known by its strong immune-inhibtory property. We examined if HLA-G plays a role as a mediator of MSC induced immunosuppression. The expression of HLA-G on human MSC alone and in mixed lymphocytes reaction MSC/MLR was analyzed.
We found that HLA-G can be detected by RT-PCR, immunofluorescence, and by flow cytometry using intracellular staining (52.4 % ± 3.6). Using ELISA a level of 38.7 ± 5.2 ng/ml of soluble HLA-G was detected in the culture supernatant. HLA-G protein expression level is constitutive and is not modified upon stimulation by allogenic lymphocytes in MSC/MLR. We analysed the functional role of HLA-G protein expressed by MSC using anti HLA-G blocking antibody (87G). We found that blocking HLA-G protein can restore lymphocytes proliferation in MSC/MLR (25.4%, P= 0.04).
Our findings provide evidence supporting the immunosuppressive properties of MSC and the role of HLA-G. It emphasizes the potential application of MSC as a relevant therapeutic candidate in transplantation.
A Novel Translocation (4;12)(q11;q13) in an Acute Promyelocytic Leukemia-Like Case Lacking RARα Rearrangements.
Authors: Qiu Hui Y. , Xue Yong Q. , Pan Jin L. , Wu Ya F. , Wang Yong , Shen Juan , Zhang Jun 
Published: October 2019

Abstract
We report a novel chromosomal translocation t(4;12)(q11;q13) in a 37-year-old male with acute promyelocytic leukemia (APL)-like morphologic changes but lacking RARα rearrangements. His blast cells had some morphologic features evocative APL such as heavy azurophilic granules, bundles of Auer rods except regular round or oval nuclei. Immunophenotype of blast cells showed positivity for CD13 and CD33, and negativity for CD34 and HLA-DR which were compatible with the diagnosis of APL. Disseminated intravascular coagulation (DIC) was present. Chromosome study of BM cells showed that 29 out of 35 metaphases had a consistent karyotype of 46, XY, t (4;12)(q11;q13) which was confirmed by chromosome painting with whole chromosome paint probes 4 and 12. However, fluorescence in site hybridization, reverse transcriptase-polymerase chain reaction (RT-PCR) and multiplex RT-PCR did not demonstrate any evidence for RARα rearrangements including PML-RARα, PLZF-RARα, NuMA-RARα, NPM-RARαand STAT5b-RARα fusion genes. Sequential treatment with arsenic trioxide and chemotherapy had no effect, he died of bleeding due to DIC. In view of the fact as mentioned above, this case should be rated as transitional M2–M3.
Retroviral Insertional Mutagenesis Screen in a C/EBPalpha Proliferative Genetic Background.
Authors: Hasemann Marie S. , Sørensen Annette B. , Pedersen Finn S. , Nerlov Claus , Porse Bo 
Published: October 2019

Abstract
The CCAAT enhancer binding protein alpha (C/EBPalpha) transcription factor plays a key role in the regulation of growth and differentiation of the granulocytic lineage in the hematopoietic system. Consistently, mice lacking C/EBPalpha have no mature neutrophils and die within a few hours after birth. In contrast, homozygous knockin mice in which the wild type Cebpa allele has been replaced with a mutant allele (BRM2) deficient in repressing the activity of E2F family members, are viable. At 8 weeks of age these animals display myeloid dysplasia with absence of neutrophil granulocytes. Strikingly, in older BRM2/BRM2 knockin mice the myeloid dysplastic phenotype progress into other myeloid malignancies such as myeloid proliferative syndrome and acute myeloid leukemia. These findings strongly suggest that secondary mutations in other loci must occur during the phenotypic progression.
In order to identify genes that cooperate with C/EBPalpha in the development of leukemia in BRM2/BRM2 mice a so-called retroviral insertion mutagenesis screen was performed. Inbred newborn BRM2/BRM2 and wildtype mice were injected with SRS19-6 retrovirus and when disease is evident the mice are euthanized and analyzed. As expected the BRM2/BRM2 mice have a shorter latency than wildtype mice (182 vs. 260 days). The mice have enlarged spleen, thymus, and lymph nodes and were further characterized by histology, flow cytometry and Southern blotting in order to determine the hematopoietic phenotypes. Most abundantly was the AML-like phenotype, but also T-cell lymphomas are developing. Finally, the loci carrying retroviral insertions loci are identified through a splinkerette-aided PCR strategy.
This study provides a better understanding of the genes involved in the development of myeloid leukemia.
Deregulation of c-Myc in Primary Effusion Lymphoma by KSHV LANA.
Authors: Bubman Darya , Guasparri Ilaria , Cesarman Ethel 
Published: October 2019

Abstract
Primary effusion lymphoma (PEL) is a rare subtype of non-Hodgkin’s lymphoma, which is associated with infection by Kaposi’s sarcoma herpesvirus (KSHV/HHV-8). PELs lack expression of B-cell markers and rearrangements in common oncogenes. c-Myc is a transcription factor that plays an important role in cellular proliferation, differentiation and apoptosis. Lymphomas frequently have deregulated c-Myc expression due to chromosomal translocations, amplifications or abnormal stabilization. However, no structural abnormalities were found in the c-Myc oncogene in PELs. Given that c-Myc is often involved in lymphomagenesis, we hypothesized that it is deregulated in PELs. We report that PELs have abnormally stable c-Myc protein. Normally, the turnover of c-Myc protein is stringently regulated by post-transcriptional modifications, which include phosphorylation of c-Myc threonine 58 (T58) by glycogen synthase-3 beta (GSK-3 beta). Our data show that the impaired c-Myc degradation in PELs is associated with drastic underphosphorylation of c-Myc T58. The KSHV latency-associated nuclear antigen (LANA) is responsible for this deregulation. Overexpression of LANA in HEK 293 cells leads to abnormal stabilization of c-Myc protein and inhibition of GSK-3 beta-mediated phosphorylation of c-Myc T58. LANA also increases c-Myc protein, but not RNA levels when overexpressed in primary B cells. Conversely, when LANA is eliminated from PELs using RNA interference, there is a post-transcriptional effect leading to a reduction in c-Myc protein. The ability of LANA to prolong the half-life of c-Myc might play an important role in the pathobiology of PEL.
Expression of Multidrug Resistance 1, Lung Resistance Protein and Breast Cancer Resistance Protein Genes in Chronic Lymphocytic and Chronic Myelogeneous Leukemia.
Authors: Ural A. Ugur , Iseri Ozlem Darcansoy , Mutlu Pelin Kaya , Kars Meltem Demirel , Avcu Ferit , Gunduz Ufuk 
Published: October 2019

Abstract
One of the major problems in the treatment of leukemia is the development of drug resistance to chemotherapeutic agents, which is already present at diagnosis or after chemotherapy as a minimal residual disease. The resistance may be originated from genetic or epigenetic mutations during prior growth of the leukemic clone. In this study, the expressions of three multidrug resistance (MDR) genes were investigated. Expression levels of multidrug resistance resistance gene 1 (MDR1), breast cancer resistance protein (BCRP) gene, and lung resistance protein (LRP) gene expression levels were determined in peripheral blood samples from 16 cases of chronic lymphocytic leukemia (CLL) and from 23 cases of chronic myelogeneous leukemia (CML) using RT-PCR. The expression of each of these genes was then expressed as a ratio in relation to β2-microglobulin gene expression in densitometric measurements. MDR1, BCRP and LRP expression levels was detected in 56,3%, 18,8% and 50% of CLL patients, respectively, with no difference for stage or response to the treatment. Four out of 16 (25%) CLL patients expressed none of these genes. The other 25% CLL patients expressed all of these genes. MDR1, BCRP and LRP expression was detected in 47,8%, 39,1% and 30,4% of CML patients, respectively, with no difference for progression or response to imatinib mesylate. Eight out of 23 (34,8%) CML patients expressed none of these genes. Four out of 23 (17,4%) CML patients expressed all of these genes, and two out of whom rapidly progressed to acute leukemia and unresponsive to the treatment. In conclusion, MDR and LRP overexpression seems to be a frequent event in CLL and CML patients; however no conclusion can be drawn on their prognostic role and response to the treatment.
Genetic Mechanisms of Cytarabine Resistance in Leukemia: Cell Culture and Mouse Transplantation Models.
Authors: Yin Bin , Diers Miechaleen D. , Rathe Susan K. , Lowe Scott W. , Largaespada David A. 
Published: October 2019

Abstract
Chemoresistance remains a major barrier to successful clinical treatment of acute myeloid leukemia (AML). However, the underlying molecular mechanisms of chemoresistance in AML are largely unknown. Previously, we reported that loss of tumor suppressor genes, Nf1 and Trp53, can confer upon myeloid leukemia cells partial resistance to cytarabine (Ara-C), a mainstay of AML chemotherapy. Microarray gene expression profiling revealed that a group of lysosome function-related genes, and a major regulator of apoptosis, Bcl-xL, were upregulated in in vitro Ara-C resistant myeloid leukemia cell lines. We now report that the lysosome number and size is increased in resistant cells. Also, overexpression of Bcl-xL in both murine and human myeloid leukemia cell lines increased Ara-C resistance to a variant extent in different lines. Furthermore, we found that the key enzyme involved in intracellular activation of Ara-C, Cda, can do so, too, but to a less extent than Bcl-xL. Interestingly, in our Ara-C resistant AML cell lines which were retrovirally induced in BXH-2 stain of mice, we were able to detected new proviral insertion sites which may cause mutations associated with the leukemia resistance feature. We have now examined the possibility of modeling AML chemoresistance in mouse, and through this model, to identify and characterize AML relapse-related genetic abnormalities. During establishing such a model, treatment dosage, intervals, courses, routes of administration, the number of injection, food preparation and supplements have been considered for testing. Ara-C treatment caused a decrease in body weights and peripheral blood cell counts. One major lethal toxicity response, intestine deterioration, appeared to be correlated with high doses of Ara-C. We arrived at a regimen comprised of two 10-day treatment courses at 90 mg/kg/day by one intraperitoneal injection, with a 5-day break between treatment courses. This treatment course was found to give rise to 10~30% lethality due to Ara-C toxicity but it can also suppress the onset of transplanted AML disease in recipient mice for nearly two weeks compared to saline-treated mice as a control. Interestingly, the tumors collected from Ara-C treated transplants displayed higher resistance than saline treated tumors in a methylcellulose colony-forming assay. Testing of apoptosis regulatory genes for Ara-C response in primary tumors is in process.
Antiproliferative and Proapoptotic Effects of Akacid-Medical-Formulation on Hematologic Cell Lines.
Authors: Neuwirt Hannes , Salvador Christina , Wabnig Elisabeth , Tiefenthaler Martin , Culig Zoran , Konwalinka Guenther 
Published: October 2019

Abstract
We have previously shown that Akacid-medical-formulation (AMF) exerts its antiproliferative effects on various malignant solid cell lines, including those derived from prostate via downregulation of mitogen-activated protein kinases Erk 1/2 and cell cycle regulators, e.g. cdk-2, -4, cyclin E, -D1 (Neuwirt et al. 2006). However, the effect of AMF on proliferation and induction of apoptosis in malignant hematologic cell lines has not been investigated yet. Therefore, we performed 3H-thymidine incorporation assays to assess the growth inhibition of HL-60, U-937, K-562, CEM-C7H2 cells after treatment for 48 hours with various concentrations of AMF (0.3 – 100 μM). All cell lines were dose-dependently inhibited by AMF with an IC50 of about 2.1μM. As cellular growth arrest is known to be one major reason for resistance to chemotherapeutics, we investigated the effect of AMF on the cell line CEM-C7H2-6E2, in which G1/0-phase arrest can be induced by tetracyclin-regulated expression of the cell cycle inhibitor p16INK4A (Fig. 1A). Flow cytometric analysis using Annexin-V / propidium-iodide staining showed that in G1-phase arrested cells apoptosis was induced to a similar extent (LD50 of 10μM) as compared to proliferating control cells (CEM-C7H2-2C8) (Fig. 1B). In addition, we tried to find out whether AMF induces apoptosis via the caspase-9 dependent intrinsic or the caspase-8 dependent extrinsic pathway. For this purpose, two different cell lines stably transfected with tetracyclin-responsible plasmids encoding for the antiapoptotic proteins bcl-2 (CEM-C7H2-10E1) and CrmA (CEM-C7H2-2E8) were used. After 48 hours of treatment with AMF (1 – 30μM) we found that overexpression of neither bcl-2 nor CrmA could inhibit AMF-induced apoptosis compared to control cells (LD50 15μM). Data on apoptosis obtained by flow cytometry were confirmed by Western blot analysis on activated caspase-8, -9, -3, and cleaved PARP in CEM-C7H2 cells. Caspase-8 and -9 were not activated after 48 hours. However, it is interesting that downstream effectors of apoptosis, cleaved PARP and caspase-3, were strongly induced (1000 % of control at 30μM). In conclusion, our results show a potent antiproliferative effect of AMF on leukemic cell lines. Induction of apoptosis could not be inhibited either by G1-phase arrest or overexpression of the antiapoptotic proteins bcl-2 or CrmA. Although we found a substantial activation of the downstream effectors of apoptosis by AMF including caspase-3 and PARP, the upstream pathway of activation remains unclear.
Figure 1 Figure 1.
Molecular Relapse Monitoring and Treatment in Acute Promyelocytic Leukemia (APL) Patients.
Authors: Shuravina E.N. , Parovitchnikova E.N. , Misyurin A.V. , Demidova I.A. , Isaev V.G. , Savchenko V.G. 
Published: October 2019

Abstract
Molecular monitoring of PML/RARA chimeric gene expression is stated to be necessary for diagnosis of acute promyelocytic leukemia (APL), as well as for monitoring of minimal residual disease (MRD). In our study we aimed to modify leukemia treatment according to MRD results. PML/RARA expression was measured by RT-PCR. Synthesis of primers and RT-PCR were performed using recommendations of BIOMED-1 Concerted Action (1999). Sensitivity of method was estimated as 10−4 – 10−6. Fresh bone marrow (BM) aspirates were investigated before treatment, during induction and consolidation, every 2–3 months during maintenance therapy, and after completion of treatment. A total of 73 morphologically suspected APL patients were included in the study; expression of PML/RARA was found in 69 patients (94.5%), bcr1/2 type in 31 patients (45%), and bcr3 type in 38 patients(55%). MRD screening during maintenance therapy was performed in 47 PML/RARA-positive APL patients. Median follow-up was 4 years (2 mo – 8 years). If chimerical transcript was revealed in a BM sample once, this was considered probable molecular relapse, and when PML/RARA was find twice in 2–3 weeks, we considered this proven molecular relapse. Molecular relapses were revealed in 19 of 47 (40.4 %) APL patients. In 6 of 19 patients (31.5 %) relapses were proven, and in 13 (68.5%) relapses were probable. When the molecular relapses were detected, maintenance chemotherapy was changed in 3 of 13 (23%) patients with probable relapses and in 5 of 6 (83%) patients with proven relapses. None of them developed hematological relapse. Therapy was not changed in 11 patients, and 3 of 10 (30%) patients with probable molecular relapse and 1 pt with proven relapse demonstrated recurrence of the disease. Thus, 4 of 11 (36%) patients with unchanged therapy developed hematological relapse. These results are significantly different from patients with modified maintenance treatment (P=0.01). Interesting data were obtained while investigating BM samples obtained from 5 patients in whom BM sampling were performed at 2 different sites (sternal and iliac). Simultaneously performed RT-PCR of those samples showed different results in 3 of 5 (60%) patients.
Though we have a small cohort of patients, we may conclude that it is advisable to change chemotherapy when proved or probable molecular relapse is revealed., When the single positive result of chimerical transcript expression is obtained, it is important to repeat RT-PCR analysis in 2–4 weeks in order to prove the molecular relapse, and then to adjust the maintenance program. However, even when it is impossible to perform the second analysis, changing the therapy may be advisable.
Detection of Chromosomal Aberrations in CLL and Correlation with Clinical Staging.
Authors: Durak Beyhan , Akay Meltem Olga , Kaytaz Behiye , Burul Ismigul , Ozdemir Muhsin , Artan Sevilhan , Gulbas Zafer 
Published: October 2019

Abstract
Chromosome abnormalities detected in CLL are important markers for determination of treatment approaches. This study was aimed to evaluate usefulness of FISH panels for testing chromosome aberrations in CLL and to correlate the aberrations with clinical staging of disease and responses to chlorombucil therapy. Cytogenetics and FISH analyses from bone marrow samples of 67 CLL patients were performed according to standard unstimulated short term culture methods. CLL specific FISH panel included probes for 11q22.3 (LSIATM), CEP12, 13q14 (LSID13S19 and LSID13S25), 17p13.1 (LSIp53). FISH analysis was performed in all samples. The most commonly seen abnormality was 13q14 deletion followed by deletions of ATM, p53 and trisomy12. Of 19 cases with 13q14 deletion, 11 had only this abnormality whereas the others had trisomy12, p53 and ATM loci deletions (multiple abnormalities). Of 11 cases with single abnormality, 8 were at stage A, 3 was at stage C and all patients achieved benefits from chlorombucil therapy except one patient. Failed therapy response was seen in 9 patients with multiple abnormalities and the disease was at stage C (4 cases) and B (5 cases). Patients showing p53 and ATM deletions related with poor prognosis had progressive disease and no benefit could be achieved from chlorombucil therapy. Of 6 cases with trisomy 12, 4 were at early stage and response to the therapy was good but in two cases at later stage, therapy benefit could be achieved in one while no benefit was seen in the other one. The results showed that FISH analysis is more informative than karyotype analysis in CLL. Detection of p53 and ATM loci deletions is an important marker for administration of second level therapy.
The Investigation of JAK2 V617F Mutation in Chinese Patients with Hematological Malignancies.
Authors: Zhang Sujiang , Li Jianyong , Li Weida , Song Junhong , Duan Limin , Lu Mize , Qiu Hongxia 
Published: October 2019

Abstract
Myeloproliferative Diseases (MPD) are a spectrum of pathogenetically related disorders of varying clinical manifestations, characterized by neoplastic expansion of relatively mature granulocyte, erythroid, megakaryocyte, or monocyte and eosinocyte lineage cells. Recently a novel point mutation affecting the Janus tyrosine kinase 2 (JAK2 V617F) was identified as pathogenetically mechanisms by multiple competing groups. To investigate its prevalence and clinical significance in Chinese patients with hematological malignancies, we introduced Allele-specific PCR (AS-PCR) combined with sequence analysis to screen JAK2 V617F mutation.
A total of 98 Chinese MPD patients and 120 additional hematological malignancies including AML, ALL, MDS were analyzed for the JAK2 V617F mutation. 98 MPD patients were referred for PV (n=57), ET (n=18), IMF (n=12), HES (n=2) and CML (n=9). 2 ml peripheral blood samples of MPD and 5–10 ml bone marrow samples of AML, ALL, MDS at the time of initial diagnosis were obtained with informed consent and genomic DNA was isolated presently. In addition, peripheral blood samples from 20 healthy donors were also collected as control. All samples were first screened by AS-PCR. The positive samples were subsequently confirmed by sequence analysis.
The results showed that JAK2 V617F mutation was detected in 43 of 57 PV patients (75.4%), 7 of 18 ET patients (38.9%) and 5 of 12 IMF patients (41.7%). None of the AML, ALL, MDS, CML was found JAK2 V617F. There is no statistical difference of JAK2 V617F positive ratio between PV, ET and IMF. Furthermore, the mutation was not detected in the HES patient and 20 healthy controls. There is no other mutations and polymorphisms throughout exon 12 of JAK2.
To our knowledge, this is the first report of JAK2 V617F mutation in a number of Chinese patients with hematological malignancies especially BCR/ABL-negative MPD. The incidence of JAK2 V617F of our study is a little lower compared with other publications especially in PV patients. The main reason may be firstly attributed to ethnic difference. In addition, with more MPD patients introduced and more sensitive methods such as ARMS-PCR or Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) applied, more JAK2 V617F mutation will be identified.
Gene Expression Signatures Separate B-Cell Chronic Lymphocytic Leukemia Prognostic Subgroups Defined by ZAP-70 and CD38 Expression Status.
Authors: Dürig Jan , Hüttmann Andreas , Klein-Hitpass Ludger , Sellmann Ludger , Dührsen Ulrich 
Published: October 2019

Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is a heterogenous disease with a highly variable clinical course and analysis of ZAP-70 and CD38 expression on B-CLL cells allowed for identification of patients with good (ZAP-70−CD38−) and poor (ZAP-70+CD38+) prognosis. DNA microarray technology was employed to compare eight ZAP-70+CD38+ with eight ZAP-70−CD38− B-CLL cases. The expression of 358 genes differed significantly between the two subgroups including genes involved in B cell receptor signaling, angiogenesis and lymphomagenesis. Three of these genes, i.e. IRTA4/FcRH2, angiopoietin 2 and Pim2 were selected for further validating studies in a cohort of 94 B-CLL patients. IRTA4/FcRH2 expression as detected by flow cytometry was significantly lower in the poor prognosis subgroup as compared to ZAP-70−CD38− B-CLL cells. In healthy individuals IRTA4/FcRH2 protein expression was associated with a CD19+CD27+ memory cell phenotype. Angiopoietin 2 plasma concentrations were two-fold higher in the poor prognosis subgroup (p<0.05). Pim2 was significantly overexpressed in poor prognosis cases and Binet stage C. Disease progression may be related to proangiogenic processes and strong Pim2 expression.