bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Clinical Significance of Programmed Death1 Ligand-1 Expression on Leukemia Cells in Adult T Cell Leukemia Patients.
Authors: Yoshimitsu Makoto , Kozako Tomohiro , Fujiwara Hiroshi , Arima Naomichi , Tei Chuwa 
Published: April 2020

Abstract
[Purpose and background] Adult T-cell leukemia/lymphoma (ATLL) caused by HTLV-I virus infection is one of the most aggressive lymphoid malignancies with short prognosis, of which best-reported median survival by combined chemotherapy is 13 months. Even at the disease onset, opportunistic infections are frequently complicated. Thus, host immune system both against tumor and infection is considered to be severely impaired in ATLL patients. However precise mechanism underlying that immunodeficiency still remains undetermined. Recently the lines of evidence that co-inhibitory molecules;programmed death-1/programmed death-1 ligand (PD-1/PD-L1) pathway in T-cell activation seems to play a crucial role in tumor escape from host immune surveillance system have been accumulated. In this study, we investigated the expression of PD-L1 on ATLL cells (CD4+/CD25+), and as their counterparts, CD4+ lymphocytes from HTLV-I asymptomatic carriers (ACs) to define their clinical significance.
[Patients and Methods] Peripheral blood CD4+ lymphocytes from 11 ATLL patients (9; untreated, 2; progressive state), 12 ACs and 2 healthy donors (HDs) were investigated for PD-L1 expression. We simultaneously measured co-inhibitory receptor, PD-1, expression on CD8+ HTLV-I Tax-specific cytotoxic lymphocytes (anti-Tax CTLs), as a marker of immunity against HTLV-I.
[Results] PD-L1 expression was detected on CD4+ lymphocytes in 3 of the 11 ATLL patients and 2 of them were in progressive state. In contrast, none of the ACs or HDs expressed PD-L1 on CD4+ lymphocytes, while those cultured with IL-2 for 48hs transiently expressed PD-L1. Anti-Tax CTLs were detected in 6 of the 11 ATLL patients and all of those CTL expressed PD-1 simultaneously. Anti-Tax CTLs were not detected in the 2 cases in progressive state. Anti-Tax CTLs were detected in 9 of 12 ACs, and PD-1 also co-expressed on those CTLs.
[Conclusions] CD4+ ATLL cells in progressive state expressed cell surface PD-L1, but not in untreated patients or ACs. This is a striking finding that exclusively aggressive form of ATLL tumor cells expressed cell surface PD-L1, which renders tumor cells to evade from host immune surveillance systems. Nonetheless large scale studies are needed to confirm these findings. In addition, Interestingly, co-inhibitory molecule PD-1 expressed on anti-Tax CTLs in not only ATLL patients but also ACs. Because PD-1 expression of CTL means “exhausted”, our observation suggests that anti-HTLV-I virus immunity is already exhausted both ATLL patients and ACs. Further experiments are warranted to define the significance of the correlation between PD-L1 on ATLL cells and PD-1 on anti-HTLV-I CTL in ATLL patients and ACs.
Alterations of Cell Signaling Pathways in Pediatric B-ALL Patients.
Authors: Accordi Benedetta , Espina Virginia , te Kronnie Gertruuy , Liotta Lance A. , Petricoin Emanuel F. , Basso Giuseppe 
Published: April 2020

Abstract
The outcome for children with Acute Lymphoblastic Leukemia (ALL) has improved in the last few decades with current therapy resulting in an event free survival exceeding 75%. Despite of this, 25% of patients still relapse or resist to conventional therapies. Proteomics, in particular the Reverse Phase Protein Arrays (RPA), offers a new approach to study both the potential molecular mechanisms of drugs chemosensitivity versus resistance, and identify new drug targets in the patients that do not respond to therapy. RPA, which can quantitatively measure dozens to hundreds of phosphorylated kinase substrates from only a few thousand cells, can be used to profile the working state of cellular signaling pathways in a manner not possible with gene microarrays, since post-translational modifications cannot be accurately portrayed by global gene expression patterns alone. We employed RPA to study the phosphorylated cellular pathways in bone marrow aspirate samples collected prior to treatment. The phosphorylation status of 95 key signaling proteins, including pro-survival, pro-apoptotic and those involved in cell cycle control were analyzed from 120 pediatric B-ALL specimens collected at the Pediatric Oncohematology Laboratory (University of Padova) over the past 15 years. Molecular network analysis and phosphoprotein profiling were performed using commercially available software. Our results indicate that there may be a difference between the molecular networks of activated kinase substrates within leukemic patients depending on disease state, genotype, or response therapy. The discovery of the molecular mechanisms related to drug resistance could play an important role for individualizing therapy and may reveal new strategies to improve treatment outcome.
Double Minute Chromosome in Acute Myeloid Leukemia.
Authors: Amin Alpesh J. , Shaw Maria , Tadros James , Benn Howard , Maroules Michael 
Published: April 2020

Abstract
Introduction: Double minutes (dmin) are small chromatin particles that lack a centromere and represent a form of extrachromosomal gene amplification. They have been found in a variety of solid tumors but are rare in hematological malignancies. C-MYC and MLL are the most common genes amplified on the dmin in AML. Here we report a case of AML in which FISH did not demonstrate C-MYC or MLL gene amplification on the dmin.
Case Report: A seventy-seven-year-old man was diagnosed with acute leukemia. Histology and flow cytometry (blasts positive for CD13, CD33, CD34, HLA-DR, CD15, CD117 and cytoplasmic MPO) were consistent with M2 AML. Chromosome analysis revealed abnormal karyotype with hypodiploidy 42–44 and loss of chromosome 5, 7,13,17,18, and Y. Structural rearrangements involving chromosomes 2,7,11,19 and 20 in addition to 2 marker chromosomes and dmin. FISH did not reveal C-MYC gene amplification. FISH did reveal MLL gene amplification, but not on the dmin. The patient received induction chemotherapy, and expired on day 14 due to cardiac complications.
Discussion: Double minutes are a mechanism for upregulated oncogene expression. After the development of molecular technology, FISH, it became possible to identify the amplified gene. A previous review (Thomas et al. Double minute chromosomes in monoblastic (M5) and myeloblastic (M2) acute myeloid leukemia: two case reports and a review of literature. Am J Hematol 2004 Sep;77(1):55–61), found C-MYC to be amplified on the dmin in 25 cases, MLL was found to be amplified on the dmin in 5 cases, and only 3 cases in which neither gene was found to be amplified on the dmin. Double minutes are associated with advanced age at presentation, a more aggressive clinical course, poor outcome with therapy, and shortened survival. The survival in most cases is reported to be less than 12 months.
Can Fibroblasts Be Reprogrammed into Macrophages?.
Authors: Feng Ru , Graf Thomas 
Published: April 2020

Abstract
Previous work showed that B cell precursors can be reprogrammed into functional macrophages by the enforced expression of the bZip transcription factor C/EBPalpha. The efficient activation of myelomonocytic genes, such as Mac-1, required the co-operation with endogenous PU.1 (Xie et al. 2004), reflecting the fact that many myelomonocytic genes are regulated by a combination of the two transcription factors. We therefore asked: Is C/EBPa and PU.1 sufficient to convert non-hematopoietic cells into macrophages? To test this, NIH-3T3 cells were co-infected with PU.1-GFP and C/EBPa-hCD4 retrovirusesor control vectors encoding the indicators GFP and hCD4 only. Uninfected cells in the retrovirus treated cultures served as additional controls. Our results showed that ~25% of the PU.1 only infected cells express Mac-1 and that this percentage could be increased ~3 fold by co-expression with C/EBPa. In addition, most cells also expressed CD45 and some expressed F4/80 antigen. The PU.1 infected and the double infected cells, but not the C/EBPa only infected cells, also expressed a number of other myelomonocytic genes as detected by RT-PCR. These included CSF-1R (M-CSFR), GM-CSF Ralpha, Lysozyme, CD32, PYK2 as well as endogenous PU.1. The PU.1 induced reprogramming of fibroblasts required the DNA binding and transcription activation domains, but not the PEST domain of the transcription factor. To test whether the reprogrammed cells have functional macrophage properties, we generated two stable cell lines co-expressing C/EBPa and PU.1 delta PEST (wild type PU.1 is toxic in long-term cultures). These cells were morphologically altered, ingested carboxylated particles, and expressed functional Fc-gamma receptors but were unable to phagocytize antibody coated red blood cells. Remarkably, the two cells lines acquired CSF-1 dependence for growth. In accordance with this finding they exhibited a 10–15 fold reduction of CSF-1 production compared to NIH3T3 cells. The response observed was not restricted to fibroblast cell lines since both embryonic and adult fibroblasts could also be partially reprogrammed by co-infection with PU.1 and C/EBPa in that they expressed Mac-1, CD45, F4/80 and IA MHC antigens. In conclusion, enforced expression of PU.1 and C/EBPa converts fibroblasts into macrophage like cells, indicating that the combination of these two transcription factors is sufficient to regulate the majority of genes that define the myelomonocytic phenotype.
CD34 Expression and Clinical Outcome of Childhood Acute Leukemia.
Authors: Kim Nam Kyun , Won Sung Chul , Lyu Chuhl Joo , Oh Seung-Hwan , Kim Hwang Min 
Published: April 2020

Abstract
Purpose: CD34 is a surface glycophosphoprotein which is expressed on early hematopoietic stem cells. It has been thought that leukemic blast with CD34 expression was associated with poor prognosis. In this study, correlation between CD 34 expression and clinical outcome of childhood acute leukemia was explored.
Method: Seventy-six patients who were diagnosed as acute leukemia in Department of Pediatrics, Yonsei University Medical Center, Seoul, Korea were enrolled in this study. All patients were analyzed retrospectively. Myeloid markers expression in acute lymphocytic leukemia (ALL) as well as CD34 presentation in ALL and acute myeloid leukemia (AML) were analyzed. Three-year event-free-survival (EFS) was calculated according to whether those antigens were expressed or not.
Result: Fifty-eight and eighteen patients were ALL and AML, respectively. Median follow up period for all patients was 2.2 years (0.1–4.6 years). Boys and girls were 33 and 25 in ALL patients, respectively. In childhood AML, 11 and 7 patients were male and female, respectively. EFS for all patients with acute leukemia was 74.1± 5.4%. Three-year EFS of AML with or without CD34 expression was 83.3±15.2 % and 46.7±15.4 %, respectively. But, the difference was not different statistically (P=0.164, Fig. 1). In ALL, Three-year EFS was 80.7±8.9 % and 78.8±7.2 % for group of CD34 negative and positive, respectively (p=0.998, Fig. 2).
Conclusion: According to data, CD34 expression of AML in children might contribute to the survival rate, however EFS was not different statistically. We think that because of small number of cohort, the statistical results were not significantly different. In ALL, there were no statistically significant differences in EFS between CD34 expression or not.
Fig. 1. Event-free survival for acute myeloid leukemia according to CD34 presentation Fig. 1. Event-free survival for acute myeloid leukemia according to CD34 presentation Fig. 2. Event-free survival for acute lymphocytic leukemia according to CD34 presentation Fig. 2. Event-free survival for acute lymphocytic leukemia according to CD34 presentation
The Expression and Regulation of ADAMTS13 in Hematopoietic System.
Authors: Liu Fang , Bai Xia , Dai Master Lan , Dong Ningzheng , Ruan Changgeng 
Published: April 2020

Abstract
Background and Objective In 2001, VWF-cleaving protease was identified as ADAMTS13, a member of the ADAMTS family of metalloprotease. ADAMTS13 is now found as the mainly cleaving enzyme of VWF in plasma. It is synthesized predominantly in the liver, although variable expression has been observed in other issues including bone marrow. Although ADAMTS13 was detected only in platelet, not other blood cells, it is unsure what kind of situations would be when we come to immature hematopoietic cells. So we investigated ADAMTS13 in cell lines such as M07e, MEG01, HEL, k562, SHI-1, NB4, MR2 etc., to see if there is ADAMTS13 produced by blast cell of megakaryocyte and granulocyte and so on.
Methods We determined ADAMTS13 cDNA by RT-PCR and ADAMTS13 expression by flow cytometry and western blotting with some mono-clone antibodies against ADAMTS13. During all those test, we use human platelets as positive control.
Results ADAMTS13 was detectable in all those cell lines. It was highly expressed in M07e and MR2, and a bit lower in MEG01 and HEL, and much lower in k562, SHI-1 and NB4. While to our surprise, ADAMTS13 was up-regulated when NB4 was treated by 1μmol/L ATRA for 72 hours. When CD11b was rising from 21% up to 95% in NB4 after treatment, expression of ADAMTS13 followed by from 24% to 80%.
Conclusions Now we know what kinds of cells may express ADAMTS13 in bone marrow, the major part of it is localized in megakaryocyte. ADAMTS13 can highly expressed in NB4 after treatment with ARTA.
The Experience on Multiparametric Molecular Diagnostic Approach of Pediatric Acute Lymphoblastic Leukemia (ALL) in Greece.
Authors: Parcharidou Agapi , Sambani Constantina , Stavropoulou Christina , Paterakis George , Belesi Chrysoula , Papadhimitriou Stefanos , Tsitsikas Constantinos , Georgakakos Vassilios , Papadakis Vassilios , Polychronopoulou Sophia 
Published: April 2020

Abstract
The rate of success in the treatment of pediatric acute lymphoblastic leukemia (ALL) has been increased steadily during the last decades. The five years’ event free survival rate is nearly 80% for children with ALL. Attempts to boost cure rates further with the use of hematopoietic stem cell transplantation have improved for some but not all, subtypes of ALL. The best hope for continued progress lies in a better understanding of the pathogenesis, the basis of resistance to chemotherapy, and finally better organized clinical trials. The present study has been based on organizing and exploring new clinical correlations among clinical data obtained from molecular genetic profile, in vitro chemosensitivity and genetic polymorphisms of detoxifying enzymes. During the last 3 years 43 newly diagnosed ALL patients, 27 boys and 16 girls, aged 23 months to 14 years old were included in this study. Bone marrow and/or peripheral blood samples were studied for karyotyping aberrations. The presence of the specific translocations t(12;21), t(9;22), t(4;11) and t(1;19) was investigated using RT-PCR and FISH. Furthermore, FISH was also used for the detection 9p deletions and MLL rearrangements. Immunophenotype of blasts and DNA index were studied by flow cytometry. In vitro chemosensitivity studies were performed by the MTT assay (ELISA).The GSTT1 genetic polymorphism (null genotype) was detected by multiplex PCR and NQ1 genetic polymorphism was detected by PCR -RFLPs. A cytogenetic/molecular result was achieved in 39/43 patients. Structural or numerical aberrations were detected in 7/39 patients. 9/39 patients were positive for the TEL/AML1 (23%) and 3/39 for the BCR/ABL fusion genes. One patient showed only one MLL allele, no patient had MLL rearrangement and 5/19 patients presented 9p deletion. A null GSTT1 genotype was observed in 5/43children (11,62%) and 14/43 patients were heterozygotes for NQ1(32,5%). 8/36 patients presented in vitro chemoresistance and 8/43 patients had DNA index &gt;1 (18,6%).In our series of patients the frequency of t(12;21) does not seem to differ significantly from the literature data. The patients who showed chemoresistance had also unfavorable prognostic markers according to cytogenetic/molecular diagnostic data or clinical characteristics. The number of the patients is low to correlate detoxifying enzymes to toxicity or response during treatment. The multiparametric diagnostic approaches in pediatric ALL seem to be of great importance in diagnosis and tailored therapy leading to high rates of cure. Our center’s effort is the optimal characterization of the pediatric ALL profile in Greece by the use of multiparametric diagnostic methods targeting a better outcome.
Bi-Allelic Involvement in AML: Examples of 16q Aberrations in Two Cases.
Authors: Howard Julie , Brazma Diana , Vaughan Beverly , Nacheva Elisabeth P. 
Published: April 2020

Abstract
Acute Myeloid Leukaemia is a heterogeneous clonal disorder of the bone marrow, which has characteristic acquired chromosomal abnormalities. Cytogenetic rearrangements of chromosome 16 have a well-established association with AML subtype M4Eo although they have also been described in other AML FAB types. Patients with rearrangements involving CBFb (16p13) and MYH11 (16q22) are associated with a more favourable prognosis. We describe two cases of AML where both homologues of chromosome 16 are involved in separate rearrangements; this to our knowledge has not been reported previously. Further more both patients have had previous treatment and thus the abnormality of the second homologue of chromosome 16 is thought to be treatment related, in-spite of which they have responded well and are both currently in remission. In case 1 (male, AML M5a) it is chromosome16 at band q22 that is involved, inv(16)(p13q22) and t(16;17)(q22;q11), while in case 2 (female, t-AML) chromosome 16 band q24 appears rearranged in der(16)t(8;16)(q1?3;q24) and t(16;21)(q24;q22). Extensive molecular cytogenetic investigations including mapping are presented and candidate gene aberrations discussed.
Differences in Proteasome Activity in Plasma of Patients with Hematologic Malignancy.
Authors: Ostrowska Halina , Kloczko Janusz , Klimiuk Miroslawa , Sokolowski Jaroslaw , Blusiewicz Eliza 
Published: April 2020

Abstract
The proteasome-dependent proteolytic pathway plays a crucial role in pathogenesis of hematologic malignancy, and as such it is a novel target for therapeutic drugs. High proteasome levels measured by conventional technique ELISA have been demonstrated in human malignant cells and in the plasma of patients with some hemopoietic malignancies and solid tumors. The objective of this study was to evaluate the activity of the 20S proteasome in the plasma from patients with hematologic malignancy.
The study included normal subjects (n=15) and untreated patients with acute lymphoblastic leukemia (ALL, n=13), acute myeloblastic leukemia (AML, n=21), multiple myeloma (MM, n= 26) and chronic lymphocytic leukemia (CLL, n=30). The 20S proteasome activity in plasma (50μg of protein) was assayed spectrofluorimetrically using synthetic peptide substrate, Suc-Leu-Leu-Val-Tyr-AMC (100μmol/L) in the presence of 0.03% SDS that selectively activates the chymotrypsin-like activity of 20S proteasome. Enzyme units (U) were expressed as pmol AMC/min.
The average activity of 20S proteasome in plasma from normal subjects was found to be 4.42 ± 0.56U/mg. The highest activity of the 20S proteasome was found in plasma from patients with ALL with a median of 55,5 U/mg (range 10.5–225.6 U/mg, p&lt;0.01). High proteasome activity was also observed in plasma from patients with AML with a median of 28.6 U/mg (13.0–64 U/mg; p&lt;0.01). In patients with MM, the average plasma proteasome activity was 11.1 ± 3.1 U/mg (range 8–18.8 U/mg; p&lt;0.01). In contrast, in patient with CLL plasma proteasome activity was similar to that in normal subjects (4.2± 1.1; p=0.05). There was a significant positive correlation with plasma lactic dehydrogenase activity (LDH) (a global marker of cell lysis), but not with white blood cell (WBC) and blast cell counts in all the examined groups of patients.
The obtained results indicate the presence of high level of 20S proteasome in the plasma of patients with ALL, AML and MM, but not with CLL, which has potential chymotrypsin-like activity.
DEAH-Box Splicing Factor Gene, Prp16 Amplification in Acute Myeloid Leukemia.
Authors: Ma Yupo , Barnett Tammy , Chai Li , Yang Jianchang , Alipio Zaida , Pei Lin , Amin Hesham M. , Fink Louis , Di Chunhui , Yan Hai , Ward David 
Published: April 2020

Abstract
Aberrant mRNA splicing has been observed frequently in solid tumors and shown to play a functionally significant role in tumorgenesis. Here we demonstrate that the DEAH-Box splicing factor gene, Prp16, is amplified in fresh human acute myeloid leukemia (AML) and in established AML cell lines. Prp16, an RNA-dependent ATPase required for pre-mRNA splicing, maps to chromosome 16q22, a region frequently altered in AML. Amplification of the Prp16 gene was initially detected using digital karyotyping, a powerful technique for analyzing genome-wide alterations in DNA copy number and verified by quantitative real-time PCR. Analysis of mRNA expression profiles revealed that Prp16 transcripts were present at high levels in 22 of 39 cases of AML (56%) and in both AML cell lines examined. There was a strong correlation between gene amplification and mRNA expression levels. To our knowledge, this is the first demonstration linking aberrant expression of a splicing enzyme to leukemogenesis. The classification and clinical outcome of the AML cases is being correlated with the presence or absence of Prp16 amplification. The identification of leukemia-specific splicing event(s) may provide a novel target for therapeutic intervention.