bluuebunny/crossref_metadata_2025_split huggingface dataset:
Usefulness of Functional Image Analysis Using Positron Emission Tomography for the Discrimination between Hypoplastic Myelodysplastic Syndromes and Aplastic Anemia.
Authors: Takatoku Masaaki , Sato Toshihiko , Hatano Kaoru , Omine Ken , Mori Masaki , Muroi Kazuo , Nagai Tadashi , Ozawa Keiya 
Published: April 2020

Abstract
Myelodysplastic syndromes (MDS) and aplastic anemia (AA) are classified bone marrow failure syndromes. Their clinical and pathological features partly overlap. It is sometimes difficult to distinguish hypoplastic MDS from AA by using current common diagnostic methods such as bone marrow biopsy and chromosome analysis. Although magnetic resonance image (MRI) is useful for diagnosis of MDS with hypercellular bone marrow, it is difficult to discriminate between hypoplastic MDS and AA using this method because the high intensity patterns on T1-enhanced images are similar in these disorders. Quantitative imaging with fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) has been recognized as a useful examination for discrimination between benign and malignant regions in various conditions. Since the decrease in FDG uptake at the late phase is much slower in a malignant region than in a benign region, dual time point imaging provides more accurate information than does conventional single time point scanning. In this study, we investigated the usefulness of dual time protocol FDG-PET in differential diagnosis of hypoplastic MDS and AA. Seventeen patients (7 with typical MDS (RA), 4 with hypoplastic MDS, 6 with AA) and 30 healthy adults agreed to participate in this study. Bone marrow biopsy, FDG-PET, MRI and CT were carried out. Spine, femur and iliac lesions were detected by their increased FDG uptake at 60 and 120 min after injection of 0.12 mCi/kg of FDG. The mean lesional standardized uptake values (SUVmean) at 60 and 120 min after injection of FDG were determined. While the median SUVmean of normal lumbar regions at 60 min was 1.123 ± 0.219, that of MDS cases was 2.06 (range, 1.62–2.32). At 120 min, the median SUVmean of normal cases was 1.180 ± 0.119, whereas that of MDS cases was 2.44 (range, 2.08–2.98). It is noteworthy that the SUVmean of four hypoplastic MDS cases was also high (ranges: 1.96–2.23 at 60 min and 1.97–2.52 at 120 min), suggesting that bone marrow in hypoplastic MDS has a hyper-metabolic state of glucose like other malignant disorders. We also observed a patchy hot area, which may be a visualization of ineffective hematopoiesis, throughout the spine image of hypoplastic MDS. In contrast, the SUVmeans at both time points of four AA cases were not different from those in normal control cases. These results suggest that functional imaging analysis using an FDG-PET dual time protocol enables discrimination between hypoplastic MDS and AA, in which MR images similar intensities.
Detection of sjTRECs within Peripheral Blood T Cells Evaluation of Recent Thymic Output Function in Myelodysplastic Syndrome.
Authors: Du Xin , Li Yang-qiu , Weng Jian-yu , Lu Ze-sheng , Guo Rong , Wu Sui-jing , Zhong Li-ye , She Miao-rong , Lin Wei , Huang Zi-lun 
Published: April 2020

Abstract
Objective
The myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal hematopoietic stem cell disorders, while, immunological abnormalities are frequently observed in patients with MDS. Several reports revealed that about 10% of MDS patients have clinical autoimmune disorders like skin vasculitis, rheumatic disease, or autoimmune hemolytic anemia. Furthermore, serological immunological abnormalities like hyper- or hypogammaglobulinemia, positivities of antinuclear antibody, positivities of direct Coombs test, or inverted CD4/8 ratios were found in 18–65% of patients with MDS. Recently immunosuppressive therapies including prednisolone, antithymocyte globulin, and cyclosporin A (CsA) are used to treat cytopenia in some patients with MDS. But it isn’t very clear the immunosuppressive mechanism in MDS and the value of the treatment. To analyze the content of signal joint Tcell receptor excision DNA circles signal joint T cell receptor excision DNA circles(sjTRECs) within peripheral blood mononuclear cells (PBMCs), thereby to infer the level of naive T cells and the recent thymic output function in patients with myelodyspoastic syndrom. Methods Quantitative detection of sjTRECs in DNA of peripheral blood mononuclear cells from 13 normal individuals and 8 patienets were performed by real-time polymerase chain reaction (PCR) and TaqMan technique.
Results
The median value of sjTRECs copies P1 000 PBMCs was 4.37±3.64 in normal individuals whereas it was1.07 ±1.40 copies P1 000 PBMCs in myelodysplastic syndrom patients (P &lt;0. 05). Conclusions MDS Patients decrease in recent thymic output function.
Analysis of the T-Cell Receptor Repertoire in the Study of Immunological Abnormalities in Myelodysplastic Syndromes (MDS).
Authors: Du Xin , Li Yangqiu , Weng Jianyu , Zhong Liye , Chen Shaohua , Yang Lijian , Lu Zesheng 
Published: April 2020

Abstract
Introduction
The myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal hematopoietic stem cell disorders, while, immunological abnormalities are frequently observed in patients with MDS[1]. Several reports [2,3]revealed that about 10% of MDS patients have clinical autoimmune disorders like skin vasculitis, rheumatic disease, or autoimmune hemolytic anemia. Furthermore, serological immunological abnormalities like hyper- or hypogammaglobulinemia, positivities of antinuclear antibody, positivities of direct Coombs test, or inverted CD4/8 ratios were found in 18–65% of patients with MDS. Recently immunosuppressive therapies including prednisolone, antithymocyte globulin, and cyclosporin A (CsA) are used to treat cytopenia in some patients with MDS. We reported four patients with MDS. Rearrangements of the TCR-beta genes were seen in these patients using RT-PCR and Genescan analysis (CDR 3 length analysis). Also they had skewed TCR usages using TCR repertoire analysis.
Methods
Two patients with refractory anemia(RA), two with refractory anemia with blasts(RAEB). four males from 41to 68 yearsold. Complementarity determining region 3(CDR3) of TCR Vβ with 24 variable region gene was amplified in peripheral blood mononuclear cells, which were drawn from five patients with myelodysplastic syndromes (MDS) using RT-PCR, to observe the expression of TCR Vβreceptoroire T cells, the PCR products were further analyzed by genescan to evaluating clonality of T cells (CDR 3 length analysis), and compare results with age-matched healthy donors and patients with graft versus host disease(GVHD).
Results
We found a significantly higher number of skewed Vb profiles in the MDS and GVHD patients compared with donors. In peripheral blood T cells, Only 2-11 Vb subfamily T cells could be identified in MDS patinets, clonal expansion T cells could be found in Vb1, 3,13, 14 and 21 subfamilies.
Disussion
In this study, we evaluated the total T-cell repertoire of 4 MDS patients using gendscan analysis to look for evidence of T-lymphocyte clonality. This analysis showed that all 4 patients exhibited extensive skewing of their TCR spectratypes, suggesting clonal or oligoclonal T-cell expansions. Epperson [10]also reported the same results. As we know, acute and chronic graft-versus-host disease(GVHD) is a major complication after allogeneic bone marrow transplantation. GVHD is mediated by T cells that are derived from the BM graft. In this study, we compared the results of GVHD patients with that of MDS patients, and found that these two groups patients show TCR Vb skew distribution and clonal expansion. These findings provide further evidence that T cell mediated immune processes are a feature of MDS.
Anti-Tumor Activity of a Novel Bcr-Abl/Lyn Dual Inhibitor, NS-187, in Murine CNS Ph+ Leukemia Models.
Authors: Yokota Asumi , Kimura Shinya , Oyama Tatsuya , Ashihara Eishi , Naito Haruna , Urasaki Yoshimasa , Ueda Takanori , Terui Yasuhito , Hatake Kiyohiko , Hirabayashi Kazuko , Maekawa Taira 
Published: April 2020

Abstract
The penetration of imatinib mesylate (Gleevec™) into the central nervous system (CNS) is poor. Hence the CNS becomes a sanctuary site for patients who are on prolonged imatinib therapy. P-glycoprotein (P-gp) plays an important role in limiting the distribution of imatinib to the CNS, and it is well known that imatinib is a substrate of P-gp. We have recently identified a specific dual Bcr-Abl/Lyn inhibitor, NS-187, which can override imatinib-resistance. NS-187 was 25–55 and at least 10 times more potent than imatinib in vitro and in vivo, respectively. The purpose of this study was to investigate whether NS-187 can inhibit the growth of Ph+ leukemic cells in the CNS. In our preliminary pharmacokinetic study, the intracranial concentration of NS-187 was 10% of its serum concentration, suggesting the involvement in P-gp. To determine whether NS-187 is effluxed by P-gp, we examined the growth-inhibitory effects of NS-187 alone and in combination with a P-gp inhibitor, verapamil or cyclosporin A, on K562 cells and on a multidrug-resistant (MDR) K562/D1-9 cell line overexpressing P-gp. The K562/D1-9 cell line was 10 times more resistant to NS-187 than the parental K562 cell line, and P-gp inhibitors abolished this resistance, indicating that the action of NS-187, like that of imatinib, is affected by the P-gp-related MDR system. Even though NS-187 was found to be a substrate for P-gp, it inhibited the growth of K562/D1-9 cells at a concentration which could be achieved in the brain. we therefore tested the anti-tumor effects of NS-187 in murine CNS leukemia models. mice were inoculated into right cerebral ventricle with 1×105 BaF3/wt bcr-ablGFP cells (Balb/c-nu/nu mice) or 1×106 K562GFP cells (NOD/SCID mice). Five days after inoculation, mice were randomized into groups of 4 and orally administrated twice a day with vehicle, imatinib or NS-187 for 14 consecutive days. Sixteen days after inoculation, three mice from each group were sacrificed and their brains were examined under a fluorescent stereoscopic microscope. NS-187 inhibited the proliferation of leukemic cells in the brain, whereas imatinib did not. Moreover, NS-187 significantly prolonged the survival of the mice in a dose-dependent manner in both murine models compared with imatinib (Figure). In conclusion, NS-187 can inhibit Ph+ leukemic cell growth in the CNS in spite of efflux of the compound by P-gp.
Figure Figure
Inherited Thrombophilias in Essential Thrombocythemia and Polycythemia Vera Are Not an Additional Risk Factor for Thrombosis.
Authors: Cacciola Emma , Cipolla Antonio , Di Francesco Ernesto , Giustolisi Rosario , Cacciola Rossella R. 
Published: April 2020

Abstract
Patients with essential thrombocythemia (ET) and polycythemia vera (PV) have a high risk of thrombosis. Inherited thrombophilias are established risk factors for thrombosis. To determine if inherited thrombophilic abnormalities might contribute to the thrombotic risk in ET and PV, we investigated antitrombin III (ATIII), protein C (PC) and protein S (PS), polymorphisms of clotting factors V (FVQ506) and II (G20210A) and methylenetetrahydrofolate reductase (MTHFR) mutation in patients with ET and PV and we tried to correlate these results with prothrombotic markers such as bthromboglobulin (bTG), platelet factor 4 (PF4), prothrombin fragment 1+2 (F1+2) and d-dimer (DD). In addition, we analysed a possible association between inherited coagulopathy and thrombosis. ATIII, PC and PS antigen levels were assayed by ELISA as well as bTG, PF4 and F1+2. PCR-based assays were used for FVQ506, prothrombin and MTHFR. DD was measured by immunoturbimetric latex agglutination. The study involved 101 patients diagnosed with ET (71 patients, 30 men, 41 women, mean age 62.3 years) or PV (30 patients, 21 men, 9 women, mean age 65.4 years) according to conventional criteria. The duration of disease ranged between 1 to 12 years. Of 101 patients, 53 (52,47%) were non-carriers of mutations and among these 24 (45.28%) experienced thrombosis, 48 (47.52%) were carriers and 27 (56.25%) had thrombosis. With reference to carriers, 9/48 (14.58%) had combined mutations such as PC and PS (2/9), PS and G20210A (1/9), PC and MTHFR (4/9), G20210A and MTHFR (2/9). The prevalence of studied defects was 31.25% (15/48) and 18.75% (9/48) for deficiency of PC and PS, respectively, 8.33% (4/48) and 14.58% (7/48) for heterozygosity of FVQ502 and G20210A, respectively, and 45.83% (22/48) for MTHFR mutation. Deficiency of ATIII was not present. All patients had higher bTG, PF4 and F1+2 (365±654 IU/ml, 133±64 IU/ml and 2.6±2.5 nmol/L, respectively) than to controls (23.8±9.1 IU/ml, 5.5±2.8 IU/ml and 0.60.2 nmol/L, respectively) (p&lt;0.0001) and normal DD (144±92 ng/L). No correlation between studied polymorphisms genetic and bTG, PF4, F1+2 and DD was found as well as with thrombotic complication. The frequency of thrombosis was not statistically different between non-carriers and carriers. Based on these data, it appears that inherited thrombophilias are not an additional risk factor for thrombosis in ET and PV and that the thrombotic phenomena are due to the abnormal platelet function that is part of the myeloproliferative disorders.
Identification of Bcr/Abl Point Mutations Conferring Resistance to the Abl Kinase Inhibitor AMN107 by a Random Mutagenesis Study.
Authors: Ray Arghya , Cowan-Jacob Sandra , Manley Paul W. , Mestan Jurgen , Griffin James D. 
Published: April 2020

Abstract
The development of kinase inhibitors such as imatinib that block the Bcr/Abl tyrosine kinase has significantly improved chronic myeloid leukemia (CML) therapy. However, patients with advanced disease often develop resistance to imatinib due to the emergence of clones with point mutations in the tyrosine kinase domain. AMN107, a novel second generation inhibitor of Bcr/Abl (Weisberg et al., Cancer Cell, 7:129, 2005) is currently in Phase 2 clinical trials and shows significant clinical activity in some patients with imatinib-resistant CML. However, it is possible that resistance to AMN107 could occur through the emergence of new Bcr/Abl point mutations, and here we report the results of a random mutagenesis study to identify Bcr/Abl mutants selected for resistance to AMN107. A library of mutations was generated in the target gene by propagating a native BCR-ABL-GFP retroviral construct through a bacterial strain deficient in a DNA repair pathway. Murine Ba/F3 cells were then transfected/infected with the mutated vector and subsequently the cells were selected for the ability to proliferate in the presence of AMN107 (0.125–0.5 μM). The Ba/F3 cells expressing native Bcr-Abl did not grow under these conditions. Single cell clones were expanded and a total of 60 individual colonies were isolated for which BCR/ABL was sequenced. Twenty colonies had single point mutations located in the kinase domain of the BCR-ABL gene. The rest had multiple point mutations and were not considered for further analysis. The point mutants identified in this way were all validated by preparing the corresponding Bcr-Abl cDNA using site-directed mutagenesis, generating a new mutant-Bcr-Abl/Ba/F3 cell line and testing for resistance to both AMN107 and imatinib. The mutant cell lines confer varying degrees of resistance to AMN107, from 5- to 400-fold. The mutant variants identified in this study included 15 novel mutations and 5 known imatinib-resistant mutations that have previously been identified in CML patients. These latter mutations included, T315I, which similar to imatinib, showed maximum resistance against AMN107 (~50% survival at 10 μM concentration). Interestingly, the majority of novel AMN107-resitant mutants were also found to be resistant to imatinib and the rank order was highly correlated to the rank order of resistance to AMN107. These data may be helpful in providing insights into the mechanism of acquired resistance of Bcr-Abl to small molecule inhibitors and are likely to predict some of the resistance mutations that may be observed in the clinic.
Myeloproliferative Disease in a Mouse Model of Down Syndrome.
Authors: Mundschau Gina , Jilani Sarah , Liu Hui , Davis Elizabeth , Le Beau Michelle , Crispino John 
Published: April 2020

Abstract
Trisomy 21 causes human Down syndrome (DS), a heterogeneous group of phenotypes including marked predisposition to leukemia. Children with Down syndrome are 500 times more likely to develop acute megakaryoblastic leukemia (AMKL) than other children. Furthermore, non-DS children with AMKL often have acquired trisomy 21 in their leukemic clones, suggesting that trisomy 21 in hematopoietic cells contributes to leukemic transformation. To better understand the impact of trisomy 21 on blood cell homeostasis and leukemia, we studied hematopoiesis in the Ts65Dn mouse model of Down syndrome. Ts65Dn mice harbor a segmental trisomy for mouse chromosome 16, homologous to human chromosome 21, and display many of the phenotypes associated with human DS, including craniofacial anomalies and learning deficits. To define the hematopoietic parameters for this strain, we performed monthly complete blood counts for a cohort of trisomic mice and their disomic littermates and discovered the development of progressive thrombocytosis and mild anemia in trisomic animals. Increased numbers of CD41+ megakaryocytes with lower modal ploidy were detected in the bone marrow and spleen of Ts65Dn mice as early as three months of age. Over time, expansion of the megakaryocyte population was accompanied by a decrease in TER119+ cells in the bone marrow, myelofibrosis, splenomegaly, and extramedullary hematopoiesis. Colony forming assays demonstrated increased colony forming ability in the spleens of trisomic mice along with variable decreased hematopoiesis in the bone marrow. Further, characterization of stem cells in the bone marrow indicated a hyperproliferative stem cell population. Importantly, the mice did not develop malignant leukemia by the age of 18 months and no mutations were found in the blood transcription factor GATA1, which is commonly affected in human AMKL. While Ts65Dn mice do not develop the AMKL seen in humans with DS, our results indicate that, trisomy 16 can cause hyperproliferation of the myeloid lineages, extramedullary hematopoiesis, and bone marrow fibrosis in mice. Additionally, these results suggest that trisomy 21 in humans may initiate a similar process in hematopoietic stem cells, which may contribute to leukemogenesis. Unexpectedly, this phenotype also bears significant resemblance to the human myeloproliferative disease chronic idiopathic myelofibrosis (CIMF). These findings may provide insight into the origins and progression of human myeloid diseases, including AMKL and CIMF.
Decitabine: Where Is the Target?.
Authors: Tamm Ingo , Sattler Nicole , Wagner Mandy , Lübbert Michael , leCoutre Philipp , Dörken Bernd , Schmelz Karin 
Published: April 2020

Abstract
The methylation inhibitor 5-Aza-2′-deoxycytidine (decitabine) has therapeutic efficacy in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Using microarray analysis, we investigated global changes in gene expression after decitabine treatment in AML. In the AML cell line OCI-AML2, decitabine induced the expression of 81 out of 22 000 genes; 96 genes were downregulated (X2-fold change in expression). RT-PCR analysis of 10 randomly selected genes confirmed the changes of expression in AML cells. Similar results were obtained with primary AML and MDS cells after treatment with decitabine ex vivo and in vivo, respectively. In contrast, significantly fewer changes in gene expression and cytotoxicity were detected in normal peripheral blood mononuclear and bone marrow cells or transformed epithelial cells treated with decitabine. Interestingly, only 50.6% of the induced genes contain putative CpG islands in the 5′ region. To further investigate the significance of promoter methylation in the induced genes, we analyzed the actual methylation status of randomly selected decitabine-inducible genes. We detected hypermethylation exclusively in the 5′ region of the myeloperoxidase (MPO) gene. DNA methylation inversely correlated with MPO expression in newly diagnosed untreated AML patients (P&lt;0.004). In contrast, all other analyzed decitabine-inducible genes revealed no CpG methylation in the promoter region, suggesting a methylation-independent effect of decitabine.
The Use of the PFA-100® and Thromboelastograph in Assessing Haemostasis and Drug Efficacy in Essential Thrombocythemia.
Authors: Cheung Betty Y.Y. , Parmar Kiran , Yadegarfar Ghasem , Hunt Beverley J. , Harrison Claire N. 
Published: April 2020

Abstract
Introduction. Essential thrombocythemia (ET) is associated with an increased risk of thrombosis. Many patients are treated with anti-platelet agents and/or cytoreductive therapy in order to minimise the risk but some still develop thrombotic events. The PFA-100® is an effective near-patient platelet function analyser and the Thromboelastograph (TEG®) Haemostasis Analyser assesses global haemostasis in whole blood. The usefulness of these technologies has not been fully evaluated in ET.
Patients and Methods. We compared results from the PFA-100® and TEG® in 25 ET patients and 19 controls. Overall the ET patients comprised 18 females and 7 males, their median age was 55 yrs, median platelet count 423x109/l. Twenty-two were on anti-platelet therapy (21 on 75mg aspirin, 1 on aspirin and clopidogrel). Seventeen patients were receiving cytoreductive therapy (10 hydroxyurea, 2 a-interferon, 4 busulphan and 1 busulphan with anagrelide). Eight patients had previous thrombosis (4 transient ischaemic attacks, 1 stroke, 1 central retinal vein occlusion, 2 multiple events [1 stroke, erythromelalgia and digital ischaemia; 1 central retinal vein occlusion and digital ischaemia]) and 2 had haemorrhagic events.
Results. All had normal INR, APTT and fibrinogen levels. Using the PFA-100®, Collagen/ADP closure times were prolonged in 15 (60%) patients compared to normal controls, suggesting platelet dysfunction, which included both patients with previous bleeding episodes. Twenty (91%) patients on aspirin had prolonged Collagen/Epinephrine closure times but for 2 (9%) patients these were within the normal range indicating aspirin-resistance. Both these patients had a history of thrombosis whilst on aspirin alone. Using TEG®, ET patients when compared to controls, took longer to initiate and to reach maximal clot strength (significantly longer R and K times, p=0.001, p=0.012 respectively and smaller angle, p=0.01) but the developed clot had a higher tensile strength (larger MA, p=0.024). There was a positive correlation between fibrinogen levels and MA (r=0.54, p=0.006) as well as APTT with R time (r=0.437, p= 0.029). There was no correlation between platelet count or prior history of thrombosis with any PFA-100® or TEG® variables.
Conclusion. Low-dose aspirin had biochemical efficacy as assessed by PFA-100® Collagen/Epinephrine closure times in most but not all ET patients. Further studies are necessary to assess whether Collagen/ADP times can identify those at risk of bleeding. In ET, the TEG® demonstrates slower formation but a higher tensile clot strength than controls. Larger studies are needed to fully evaluate these findings and their clinical relevance.