bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Expression Profiling of Philadelphia Chromosome(Ph) Negative CD34+ Hematopoietic Stem and Progenitor Cells of Patients with Ph Positive CML in Complete Molecular Remission during Therapy with Imatinib.
Authors: Neumann Frank , Teutsch Nathalie , Kliszewski Slawomir , Bork Simone , Brors Benedikt , Schimkus Nadine , Roes Nicole , Steidl Ulrich , Germing Ulrich , Hildebrandt Barbara , Royer-Pokora Brigitte , Eils Roland , Gattermann Norbert , Haas Rainer , Kronenwett Ralf 
Published: October 2019

Abstract
Treatment of chronic myelogenous leukemia (CML) with Imatinib has shown its efficacy and superiority to conventional therapies. Beside inhibition of BCR-ABL Imatinib also has inhibitory effects on other tyrosine kinases such as wild-type ABL, c-KIT and platelet-derived growth factor receptors (PDGF-R) alpha and beta which play a role in differentiation and proliferation of hematopoietic stem cells and are involved in DNA repair mechanisms. Inhibition of these genes might therefore result in functional disturbance of hematopoiesis or even in secondary karyotypic abnormalities. Therefore, we compared gene expression profiles of immunomagnetically isolated CD34+ bone marrow (BM) cells from six healthy volunteers with Ph negative CD34+ cells from BM of eight patients (6 males, 2 females; median age: 52 years, range: 25–68 years) with Ph positive CML in chronic phase who reached major molecular remission during imatinib therapy (400 mg/day; 28 months (range: 11–39 months)). Cytogenetics, FISH analysis and quantitative real-time RT-PCR for BCR-ABL transcripts showed that all patients were in complete cytogenetic and in major molecular remission ( ≥ 3 log reduction of the BCR-ABL/G6PDH ratio in comparison to pretreatment value). CD34+ cells (median: 5 x 105; range 2 x 105–1.8 x 106) were isolated immunomagnetically from bone marrow with a purity >98%. Total RNA (median: 550 ng; range: 130 – 990 ng) from isolated CD34+ cells was used to generate biotin-labelled cRNA (median: 7.5 μg; range: 2.6–12.3 μg). Labeled cRNA was hybridized to Affymetrix HG-Focus GeneChips covering 8793 genes representing a broad spectrum of different functional groups. For normalization and data analysis we used the statistical scripting language “R”. Raw data were normalized using a method of variance stabilizing transformations (VSN). To compare the normalized data from CD34+ cells of imatinib-treated patients and healthy volunteers we used the Significance Analysis of Microarrays (SAM) algorithm v1.21. We found that CD34+ cells of patients after reaching major molecular remission during first-line treatment with 400 mg imatinib per day showed no significantly differentially expressed genes in comparison to CD34+ cells of healthy volunteers in vivo. There was no evidence for transcriptional changes of genes involved in DNA repair or oncogenesis supporting the view that secondary chromosomal aberrations in Ph negative hematopoiesis observed during imatinib therapy might not be induced by the tyrosine kinase inhibitor itself. Our data suggest that imatinib as first-line therapy might not result in a major functional disturbance of Ph negative hematopoiesis.
Recurrent Fusion of PCM1 to JAK2 in Atypical Chronic Myeloid Leukemia and Acute Leukemia Associated with the t(8;9)(p21-22;p23-24).
Authors: Reiter Andreas , Walz Christoph , Watmore Ann , Schoch Claudia , Blau Ilona , Telford Nick , Aruliah Shilani , Yin John , Vanstraelen Danny , Barker Helen , Taylor Peter , O’Driscoli Aisling , Schlegelberger Brigitte , Berger Ute , Hochhaus Andreas , Hehlmann Rüdiger , Chase Andrew , Cross Nicholas C.P. 
Published: October 2019

Abstract
We have identified a novel, recurrent t(8;9)(p21–22;p23–24) in six patients with diverse hematological malignancies: atypical CML (n=4), secondary AML following idiopathic myelofibrosis (n=1) and pre B-ALL (n=1). Because of the involvement of several different tyrosine kinases in atypical CML, we focused our analysis on this class of gene. Initial FISH studies of one patient indicated that the janus kinase 2 gene (JAK2), located at 9p24, was disrupted. RACE-PCR was then used to identify the 8p21 partner gene as PCM1, a large centrosomal protein that contains multiple coiled-coil domains. RT-PCR and FISH analysis confirmed the fusion in this case, and also identified PCM1-JAK2 in the five other t(8;9) patients (RT-PCR and FISH, n=4; RT-PCR only, n=1; FISH only, n=1). Four different types of in-frame mRNA junction were identified, but in all cases the chimeric mRNA is predicted to encode a protein that retains several of the predicted coiled-coil domains from PCM1 and the entire tyrosine kinase domain of JAK2. Reciprocal JAK2-PCM1 mRNA could not be amplified in any patient. Clinically, 4 patients displayed CML-like hyperplasia with variable degrees of myelofibrosis and eosinophilia. Similar to typical CML, the clinical course of these patients was variable: one is alive 11 months after allogeneic stem cell transplantation, one transformed to acute leukemia 5 years after diagnosis, one died 4 days after presentation and one achieved a major cytogenetic response with interferon but died due to neurodegenerative disease and pneumonia at 7.5 years. Of the two remaining patients, one presented with secondary AML following idiopathic myelofibrosis and remains in remission 15 years after diagnosis following intensive chemotherapy and maintenance with interferon. The final patient died shortly after induction therapy for pre B-ALL. In conclusion, PCM1-JAK2 is a novel recurrent fusion gene in hematological malignancies that is likely to deregulate hemopoiesis in a manner similar to BCR-ABL. Patients with PCM1-JAK2 disease are attractive targets for signal transduction therapy.
Flow Cytometric FLT3 Expression in Acute Leukaemias Is of Diagnostic Value but Does Not Correlate with ITD / D835Y Mutation Status.
Authors: Ridler Colin , Smith Matthew , Milne Tim , Ebrahim Fatema , Macey Marion , Lister T. A. , Fitzgibbon Jude , Cavenagh Jamie D. , Agrawal Samir G. 
Published: October 2019

Abstract
High level expression of FLT3 in acute myeloid (AML) and lymphoid (ALL) leukaemias has been demonstrated by PCR techniques. This suggests that FLT3 and its ligand may play a significant role in the proliferation of leukaemic blast cells. Mutations of FLT3 are the most common genetic lesion in AML, occurring in 20–30% of cases, and confer an adverse prognosis. Flow cytometry was used as an alternative method to investigate FLT3 expression in acute leukaemia. Surface expression of FLT3 was measured in 111 bone marrow or peripheral blood samples of patients with AML or ALL using a phycoerythrin-labelled monoclonal IgG1 antibody (Coulter-Immunotech). Exons 14 – 15 and 20 were amplified from DNA extracted from mononuclear cells of 38 of the 111 samples. Internal tandem duplication (ITD) and mutation of the D835 position of the activation loop of FLT3 were identified by SSCP and direct sequencing. 82% (47/57) of AML and 81% (34/42) of B-lineage ALL samples showed ≥ 20% (range, 20 – 97%) positivity for FLT3 using flow cytometry. The Mean Fluorescence Intensity (MFI) was similar between the two groups and no significant difference was found between adult (82%) and paediatric (80%) samples. However, FLT3 expression in all 9 T-ALL samples (pro, intermediate and late subtypes) was negative with markedly reduced MFIs. Of the 25 AML cases tested for ITD/D835Y, 5 (20%) were positive (4 ITDs, 1 D835Y). Surface FLT3 expression in these 5 cases (% and MFI) was the same as in those with wild-type FLT3. No mutations of FLT3 were found in the ALL cases tested (9 B-ALL, 4 T-ALL). Of three biphenotypic leukaemias, one case with myeloid/B lineage positivity was FLT3+ by flow cytometry, while another with myeloid/T lineage markers, was FLT3−. A diagnostic conundrum was a patient with AML morphologically (cytogenetics failed): CD33+, MPO−, CD13− CD117−; T-lineage markers CD2+, CD5+, CD7+, as well as TdT+. The T-lineage specific marker cytoplasmic CD3 was negative. Only FLT3 positivity allowed a definitive diagnosis of AML to be made. In conclusion, surface FLT3 expression was present in > 80% of AML and B-lineage ALL, but in 0% (0/9) of T-ALL cases. Detection of this antigen should be included in the diagnostic immunophenotyping of acute leukaemias and is important where T-ALL needs to be excluded. Flow cytometric FLT3 expression cannot differentiate cases with or without ITD or point mutations of FLT3.
In Chronic Inflammation, There Exists an IL-6 Independent Pathway for the Induction of Hepcidin.
Authors: Rivera Seth , Gabayan Victoria , Ganz Tomas 
Published: October 2019

Abstract
The iron regulatory hormone hepcidin is presumed to be the mediator of anemia of inflammation (AI). Patients with hepatic adenomas producing large amounts of hepcidin had severe AI that resolved when the tumors were resected (Weinstein et al, 2002). Patients with disorders associated with AI and mice with acute inflammation also have increased hepcidin. However, hepcidin deficient mice do not develop hypoferremia in response to acute inflammation (Nicolas et al, 2002). We previously showed that IL-6 was necessary for the inflammatory induction of hepcidin and the development of hypoferremia in an acute turpentine model of inflammation (Nemeth et al, 2004). In order to determine whether IL-6 was necessary for the development of AI, we induced chronic peritoneal S. epidermidis abscesses in IL-6 deficient mice (19 with abscesses, 12 controls) and wild type mice with the same genetic background (16 with abscesses, 10 controls). In contrast to our previous results, both IL-6 deficient and wild type mice had a significant rise in hepcidin-1 expression (27.6 vs. 10.9-fold, respectively) but there was no difference between the groups (p=0.939 by 2-way ANOVA). Both IL-6 deficient and control mice demonstrated a similar small but significant fall in hematocrit (49% to 44% vs. 46% to 42%, p=0.538 by 2-way ANOVA) in response to the abscesses. IL-6 is not necessary for the induction of hepcidin or the development of AI in a physiological model of infection. Experiments are underway to define which other inflammatory mediators induce hepcidin.
Functional Studies of Human Bone Marrow Derived Side Population Cells.
Authors: Banu Naheed , Fabian Attila , Mulligan Richard C. 
Published: October 2019

Abstract
Rationale:
We have previously reported that staining of murine or human bone marrow with the vital dye, Hoechst 33342, identifies a rare population of dye-effluxing cells termed the “side population” (SP). In mice, competitive blood repopulation assays demonstrate that murine SP cells are highly enriched in hematopoietic stem cells (HSC). However, whether bone marrow SP cells in humans similarly possess functional stem cell characteristics is not currently known. We tested the stem cell potential of human SP cells in comparison to non-SP and AC133+ cells by transplantation of bulk or purified marrow sub-populations into irradiated NOD/SCID mice.
Methods:
Ficolled human bone marrow preparations were stained with Hoechst dye or a fluorescence-conjugated antibody against AC133. Flow cytometry was used to purify SP cells, non-SP cells designated as ‘MP”, AC133+ cells, or unfractionated mononuclear (MNC) cells. Several cell doses of each sub-population were intravenously transplanted into sub-lethally irradiated SCID/NOD recipient mice. Long-term bone marrow chimerism was analyzed 6–8 weeks post transplant to assess the SCID/NOD repopulating potential of each sub-population.
Results:
Transplantation of 1x10^6, 500, 000 and 100, 000 mononuclear cells resulted in detectable but low-level marrow chimerism 5.14%, 1.13%, 0.17%. Transplantation of same number of MP cells also resulted in low level of chemerism (3.88%, 1.20%, 0.67%) compared to SP cells. High level of chemerism was obtained with 40,000, 20,000, and 10,000 SP cells 7.09%, 1.4%, and 0.50% respectively. In comparison 40,000, 20,000, and 10,000 AC133+ cells resulted in 2.57%, 0.97, and 0.07% chimerism. To examine whether the ability of AC133+ cells to efflux Hoechst dye could better identify cells with HSC potential we transplanted 5,000 AC133+ SP cells vs. 5,000 AC133+ non-SP cells into SCID/NOD recipients and found that AC133+ cells that do not efflux Hoechst (non-SP) do not contribute to long-term marrow chimerism whereas AC133+ SP cells resulted in 0.63% chimerism.
Conclusion:
We show for the first time that human bone marrow SP cells possess functional hematopoietic stem cell characteristics. Similarly, all of the NOD/SCID repopulating activity of adult human bone-marrow AC133+ cells are found to be more enrich in the “SP”AC133+ fraction (0.1% SP) compare to total mononuclear, or “MP”AC133+ or total AC133+ cells.
Polychemotherapy in patients with primary CNS lymphoma.
Authors: Glasmacher Axel , Pels Hendrik , Helmstaedter Christoph , Deckert Martina , Kroschinsky Frank , Schackert Gabriele , Reichmann Heinz , Egerer Gerlinde , Ho Anthony , Schulz Holger , Engert Andreas , Diehl Volker , Bode Udo , Klockgether Thomas , Schlegel Uwe , Schmidt-Wolf Ingo G.H. 
Published: October 2019

Abstract
In primary CNS lymphoma (PCNSL) irradiation alone is no longer the primary treatment and combination with chemotherapy has significantly improved response and survival. However, patients treated with combined modality therapy suffer from significant neurotoxicity. Therefore, studies with chemotherapy alone have been carried out. We recently reported on a phase II multicenter trial to evaluate event-free survival, overall survival, response rate, response duration and toxicity of combination chemotherapy in patients with previously untreated PCNSL. 65 consecutive patients with histologically proven PCNSL received a total of 6 courses of combination chemotherapy with 3 different courses of chemotherapy. In patients with response these 3 cylces were repeated. The combination chemotherapy included high-dose methotrexate (MTX) (5g/m2, course 1,2,4,5) and cytarabine (ara-C) (3g/m2/d, d1-2, courses 3,6) combined with dexamethasone (courses 3–6), vinca alkaloids and ifosfamide alternating with cyclophosphamide. In addition, MTX, prednisolone and ara-C were applied via an Ommaya or Rickham reservoir in each course. The study population included 65 consecutive patients with a median age of 62 years with a range between 27 and 75. 62 patients are evaluable for response. There were 9% therapy-related deaths mainly due to neutropenic infections. Ommaya reservoir infections occurred in 12/64 (19%) and acute transient MTX related encephalopathy in 2 (subacute in 1) patients. The 5-year-survival of patients below 61 years was 75%, of patients of 61 years or older 17% (Pels H et al., J Clin Oncol21(24), 2003). The results for patients younger than 60 years could be superior to previously reported therapy regimes. The survival rates for patients above 60 years are comparable to those reported for combined radiochemotherapy. However, in contrast to radiochemotherapy permanent treatment-associated neurotoxicity is infrequent. Based on these data we constructed a follow-up trial. Patients below the age of 61 are treated as in the antecedent protocol without intraventricular therapy. Patients above 60 are treated in a protocol modified by the addition of procarbazine in 2 cycles and a maintenance therapy with procarbazine. First results of this follow-up trial will be presented.
Inv (16) Suppresses CEBPA in Acute Myeloid Leukemia by Activation of Calreticulin.
Authors: Helbling Daniel , Mueller Beatrice U. , Timchenko Nikolai A. , Betts David R. , Jotterand Martine , Meyer-Monard Sandrine , Fey Martin F. , Pabst Thomas 
Published: October 2019

Abstract
Inversion of chromosome 16 (inv(16)) results in the reciprocal chromosomal rearrangement of the CBFB and MYH11 genes and it is the hallmark of malignant cells observed in patients with acute myeloid leukemia (AML) subtype M4Eo. Alterations of structure or expression of CEBPA - a key myeloid transcription factor - have been implicated in particular subtypes of AML. To investigate the effects of inv(16) on CEBPA we conditionally expressed inv(16) in U937 cells and found that CEBPA mRNA levels remained unchanged. However, CEBPA protein and binding activity were suppressed by 100% and 71%, respectively. In parallel, analysis of patients with inv(16) (n=12) as compared to normal karyotype AML M4 patients (n=6) showed that CEBPA protein and binding activity were significantly reduced (100% and 71.6%, respectively) whereas CEBPA mRNA levels remained similar. Calreticulin, an inhibitor of CEBPA translation, was induced on mRNA and on protein level in inv(16) AML patients as well as after conditional expression of inv(16) in a U937 cell system. Furthermore, inhibition of Calreticulin by siRNA powerfully restored CEBPA levels. Our results identify CEBPA as a key target gene of the leukemic fusion protein inv(16) and suggest that suppression of CEBPA by Calreticulin contributes to the differentiation block in AML with inv(16).
Identifying Hypercoagulable States with the Overall Haemostatic Potential (OHP), a Simple Global Haemostasis Assay.
Authors: Curnow Jennifer L. , Morel-Kopp Marie-Christine , Aboud Margaret , Ward Christopher 
Published: October 2019

Abstract
Hypercoagulable states typically result in arterial and/or venous thromboembolism. Routine coagulation tests do not identify affected patients and thrombophilia testing detects, at best, 50% of patients at risk. We hypothesise that the OHP can differentiate patients with various hypercoagulable states from a control group. The OHP is a global haemostasis assay of fibrin generation and lysis in citrated plasma. It produces a number of parameters for comparison. We established reference ranges with plasmas from 100 normal donors, then assayed plasmas from 90 clinically hypercoagulable patients being tested for a lupus anticoagulant. A further 81 patients with demonstrable antiphospholipid antibodies (APLAs) were then assayed. Statistical analysis involved calculation of means, medians, standard deviations and Mann-Whitney testing. All parameters tested were significantly different in the clinically hypercoagulable group (p<0.001), demonstrating accelerated fibrin generation and markedly reduced fibrinolysis. On subgroup analysis patients with arterial (8) or venous thromboembolism (30), antiphospholipid antibodies (9), normal or complicated pregnancy (22) and autoimmune diseases (12) all showed significantly increased fibrin generation and lysis compared with controls. The subsequently tested 81 patients with APLAs also showed a significantly hypercoagulable OHP compared with controls (p<0.01). APLA positive patients with thromboembolism showed a non-significant trend to accelerated fibrin generation and delayed fibrinolysis when compared to those without thromboembolism. APLA positive patients taking warfarin showed significant hypercoagulability when compared with APLA negative patients on long term warfarin therapy (p < 0.01). The OHP clearly identifies patients with clinically hypercoagulable states, even in the presence of warfarin. Fibrin generation is increased and fibrinolysis delayed. We postulate that the OHP may identify patients at high risk of thromboembolism.
The Effect of Patient Age and Calendar Year on the Incidence of Idiopathic vs. Secondary Venous Thromboembolism (VTE): A Population-Based Cohort Study.
Authors: Kobbervig Catie E. , Heit John A. , Petterson Tanya M. , Christianson Teresa J. , Bailey Kent R. , Melton L. Joseph 
Published: October 2019

Abstract
If the observed dramatic increase in VTE incidence with advancing age is due to increased VTE risk factor exposure (i.e., secondary VTE), the incidence of idiopathic VTE should not vary with age or calendar year.
Objective: To estimate the incidence of idiopathic and secondary VTE by age and by calendar year.
Methods: Using the resources of the Rochester Epidemiology Project, we identified the inception cohort of Olmsted County, MN, residents with a first lifetime VTE during the 30-year period, 1966–1995 (n=2761). For each case, we reviewed the complete medical records in the community for 48 baseline clinical characteristics that are commonly-accepted risk factors for VTE. We categorized VTE cases as idiopathic (n=305) if no such characteristics were present; the remaining cases were categorized as secondary. Age- and sex-specific incidence rates were calculated using idiopathic or secondary VTE cases as the numerator, and age-, sex- and calendar year-specific estimates of the population of Olmsted County as the denominator.
Results: The incidence of both idiopathic and secondary VTE increased exponentially with age for both men and women (p<0.001). Over the 30-year study period, the age-adjusted incidence of idiopathic VTE was essentially constant among men (from 19.0 to 17.1 per 100,000 men-years for 1966–70 and 1990–95, respectively), but decreased markedly among women (from 18.5 to 3.6 per 100,000 woman-years for 1966–70 and 1990–95, respectively; p=0.005 for the interaction).
Conclusions: The dramatic increase in VTE incidence with age likely reflects the biology of aging, although as yet unidentified VTE risk factors cannot be excluded. The reason for the decreased incidence of idiopathic VTE over the last 30 years among women is unclear.