bluuebunny/crossref_metadata_2025_split huggingface dataset:
Response Adapted Therapy: Using FDG-PET after Chemotherapy in Order To Restrict Indication for Radiotherapy in Children and Adolescents with Early Stages of Hodgkin’s Lymphoma (HL).
Authors: Mauz-Koerholz Christine G. , Kluge Regine , Sorge Ina , Merziger Barbara , Krausse Antje , Hasenclever Dirk , Kamprad Friedrich H. , Amthauer H. , Franzius Christiane , Gruenwald F. , Sabri Osama , Hirsch Wolfgang , Korholz Dieter 
Published: April 2020

Abstract
Introduction: Radiotherapy is an important risk factor for the development of secondary malignancies in children with Hodgkin’s lymphoma. The GPOH-HD-study group tries to restrict the indication for radiotherapy after two cycles of intensive OPPA (OEPA) chemotherapy. In the GPOH-HD 95 trial 113/394 (29%) of patients with early stage HL were in complete remission (CR) after chemotherapy based on CT/MRI (=CRI) criteria. They were not irradiated and achieved an excellent 97% EFS rate at 5 yrs (Dörffel, 2003).
Objective: Our next study will drop radiotherapy also in patients with residual findings according to CRI criteria if FDG-PET converts to negative after chemotherapy as ascertained by real-time central review of all images. Here we report pilot data on the conversion rate.
Results: During the GPOH-HD 2002 Pilot study 41 patients with early stage HL received both a staging PET at diagnosis and a restaging PET after chemotherapy. Images were centrally reviewed in Leipzig. 7 patients were in CR based on both PET and CRI criteria, one patient with CRI-based CR had still a positive PET result; in a further patient PET was not completely evaluable. In 17/32 patients with residual masses PET was completely negative after chemotherapy. In one patient PET was completely negative except for one initially involved site, where it was not informative due to a local artefact; however, CRI showed local CR in this region. In summary, 27/41 (66%) patients would qualify for no radiotherapy within a PET-based response adaptation strategy in contrast to only 9/41 (22%) patients when CRI-based. Interestingly, in 13/17 patients with negative PET, a residual mass within the mediastinum was found, indicating that a PET-based strategy will prevent many patients from mediastinal irradiation.
Conclusion: A FDG-PET-based response evaluation has the prospect to avoid radiotherapy in about 60 – 70% of all patients with early stage HL. Our next trial is designed to demonstrate that this does not lead to an unacceptable increase of the relapse rate.
Incidence of Pulmonary Hypertension in Haemoglobinopathic Patients without Left Ventricular Disfunction.
Authors: Derchi Giorgio , Formisano Francesco , Lamagna Martina , Galanello Renzo , Bina Patrizio , Cappellini Maria Domenica , Fasulo Maria Rosaria , Prossomariti Luciano , Cinque Patrizia , Maggio Aurelio , Renda Disma , Cianciulli Paolo , Cogliandro Tiziana , Piga Antonio , Donato Guido , Forni Gian Luca 
Published: April 2020

Abstract
Incidence of Pulmonary Hypertension (PH) has been described in haemoglobinopathic patients, ranging from 10% to 50%, depending on age, therapy, coexisting left ventricular dysfunction (LVD) and classification of the disease. To determine whether PH can occurs in haemoglobinopathic patients in absence of overt LVD, we studied 1121 pts from 7 Thalassemia Centers in Italy(485 Thalassemia Major (TM), 458 thalassemia intermedia (TI), 178 Sickle Cell or Sickle Thalassemia Disease (SCD),49% males, mean age 30 yrs, range 13–56 yrs). In all pts echocardiographic (Echo) defined ejection fraction (EF%) was &gt; 45% and clinically overt LVD was excluded. A cross-sectional Echo study was performed in order to detect PH that was prospectively defined as mild with tricuspid regurgitant jet velocity (TRV)of at least 2.5 m per second and defined as severe with a TRV&gt; 3.5 m per second. Severe PH was observed in 9 pts, 3 with TM, in 6 with TI and in 1 patients with SCD. Mild PH occurs in 46 patients with TM, in 57 patients with TI and 11 pts with SCD. In the remaining patients, PH was in the normal range (TM 89,9%, TI 86,2% and SCD 93,2% of pts, respectively). In the whole group of hemoglobinopathic pts the incidence of PH (mild plus severe) range from 6.5 % to 13.8% of pts. PH (mild to severe) particularly affects pts with TI (13,8% of pts).
Incidence of PH in TM, TI and SCD Tricuspid Regurgitant Jet Velocity Thal Major 485 pts Thal Int 458 pts SCD 178 pts % pts TRV&lt;2.5 m/sec 436 (89.9%) 395 (86.2%) 166 (93.2%) 89% 2,5&lt;TRV&lt;3.5 m/sec 46 (9.4%) 57 (12.5%) 11 (6.2%) 10.1% TRV&gt;3.5 m/sec 3 (0.6%) 6 (1.3%) 1 (0.9%) 0.9%
Our data support a significant presence of PH in haemoglobinopathic pts (6.2 to 13,8%) even though well treated and without LVD. Considering the pathophisiology of haemoglobinopathies a specific consideration to PH in these diseases is required in order to establish a preventive and therapeutic approach.
Endogenous Platelet PF4 Promotes In Vivo Activated Protein C (APC) Generation and Survival after Lethal Lipopolysaccharide Challenge in Mice: A Potential Physiologic Role for PF4.
Authors: Kowalska M. Anna , Mahmud Shawn A. , Poncz Mortimer , Slungaard Arne 
Published: April 2020

Abstract
Pharmacologic infusion of activated protein C (APC) improves survival in human sepsis and genetically induced deficiencies in APC generation increase mortality in a murine lipopolysaccharide (LPS) model of inflammation/sepsis. We have shown that the platelet α-granule-specific chemokine, platelet factor 4 (PF4), accelerates thrombin·thrombomodulin complex APC generation in vitro and in vivo when recombinant PF4 is pharmacologically co-infused in a primate thrombin infusion model system. Hypothesizing a potential physiologic role for PF4 in stimulating APC formation in vivo, we tested whether variations in the level of endogenous platelet PF4 content affect APC generation and also influence survival in the murine LPS challenge model. To stimulate APC formation and platelet PF4 release in mice, we injected murine thrombin (80U/kg IV). Ten min after thrombin injection into wildtype (WT) C57BL/6J mice, plasma APC levels, assessed by enzyme capture amidolytic assay, increased ~10-fold over basal levels. Injection of concurrent murine PF4 (7.5 mg/kg IV) further increased APC levels 2-2.6-fold. We then studied APC generation in mice that were either completely deficient in PF4 (PF4−/−) or transgenic mice that had 6-fold normal levels of human PF4 (hPF4+), both of which had been crossbred onto the same C57BL/6J background &gt;10 times. Plasma APC generation by thrombin was 74 ± 6% in the PF4−/− mice (n = 7) compared to WT littermates (n = 15, p&lt;0.1) and increased to 178 ± 49% in hPF4+ mice (n= 22, p&lt; 0.003 compared to WT (n= 36)). Heterozygous protein C deficient (PC+/−) mice had 65 ± 21% of WT APC generation (n= 5, p= 0.05 compared to WT (n=7)), but APC generation was restored to 96 ± 6% level in hPF4+/PC+/− mice (n= 6, p= 0.88 compared to WT (n=12)). Only PF4−/− and PC+/− mice died during this thrombin injection trial with mortality rates of 30% and 58%, respectively. A lethal LPS challenge model (25 mg/kg IP) was used to test the susceptibility of mouse strains with varying PF4 levels. Significant numbers of WT and PF4−/− mice died before the end of the first day [(18 of 44 (40% mortality) and 6 of 21 (28% mortality), respectively], while none of the 19 hPF4+ animals died (p&lt; 0.001). However, after the first day, mortality did not differ significantly between the groups and final mortality for the three groups was ~70%. In summary, these studies of mice with varying platelet PF4 suggest that endogenous PF4 plays a physiologic role to stimulate APC production. Moreover, mice with high platelet PF4 content appear to be protected from both thrombin- and LPS-provoked deaths. This suggests that PF4 plays a physiologic role to enhance APC generation, that patients with high PF4 levels may be intrinsically protected from sepsis and supports the possibility that PF4 infusions in patients with sepsis may be a viable therapeutic strategy.
The CBFb-SMMHC Oncoprotein Inhibits Binding of the Runx1 Runt Domain to DNA.
Authors: Bushweller John H. , Lukasik Stephen M. , Speck Nancy A. 
Published: April 2020

Abstract
Core-binding factors (CBFs) are heterodimeric transcriptional factors consisting of a DNA-binding Runx1 (CBFα) subunit and a CBFβ subunit. Cbfβ allosterically increases the affinity of Runx1 for DNA ~2.5 fold. CBF subunits are encoded by four genes in mammals. RUNX1 (AML1), RUNX2, and RUNX3 encode for CBFα subunits, and CBFB encodes the CBFβ subunit. Homozygous disruption of either the Runx1 or the Cbfb genes in mice results in essentially identical phenotypes: midgestation embryonic lethality accompanied by extensive hemorrhaging and a profound block at the fetal liver stage of hematopoiesis. In humans, chromosomal rearrangements that disrupt the Runx1 and CBFB genes are associated with a significant percentage of leukemias. CBFβ is disrupted in acute myeloid leukemia by inv(16)(p13;q22), t(16;16), and del(16)(q22). These translocations result in the production of novel fusion proteins containing most of the CBFβ protein fused to the C-terminal coiled-coil domain from smooth muscle myosin heavy chain (SMMHC) encoded by the MYH11 gene. A knock-in of the CBFB-MYH11 allele in mice resulted in embryonic lethality with a profound block in hematopoietic development, the same phenotype observed for the Runx1 and Cbfb knockouts. We recently demonstrated that the CBFβ-SMMHC fusion protein binds to the DNA binding Runt domain from Runx1 with both higher affinity and altered stoichiometry relative to native CBFβ. We also provided NMR-based evidence for multiple sites of contact between Runx1 and CBFβ-SMMHC, proving the role of the SMMHC sequence in creating this altered affinity. Here we demonstrate that CBFβ-SMMHC inhibits DNA binding of the Runx1 Runt domain by ~6-fold for the CD4 dual-site silencer element. Cross-saturation NMR mapping on the Runt domain in complex with CBFβ-SMMHC reveals that the SMMHC portion of the oncoprotein makes contacts with β-strands 1 and 2 in the Runt domain. We propose that the inhibition of DNA-binding and increased affinity combine to mediate the dysregulation of Runx-regulated genes caused by CBFβ-SMMHC. These results also clearly suggest that targeting of the CBFβ-SMMHC protein for drug development may well be a viable approach for the treatment of the associated leukemia.
Reduction of Core Binding Factor beta (CBFβ) Dosage Blocks T Cell Development.
Authors: Guo Yalin , Talebian Laleh , Maillard Ivan , Speck Caroline L. , Pear Warren S. , Rothenberg Ellen V. , Speck Nancy A. 
Published: April 2020

Abstract
Core binding factors (CBFs) are heterodimers consisting of a DNA binding subunit (Runx1, Runx2, or Runx3) and a non-DNA binding CBFβ subunit. CBFβ increases the affinity of the Runx subunits for DNA. Embryos deficient for Runx1 or CBFβ die at midgestation with a complete failure of definitive hematopoiesis due to a block in hematopoietic stem cell (HSC) emergence. To examine the role of core binding factors at later stages of hematopoiesis, we generated a hypomorphic Cbfb allele (Cbfbrss), that when carried over a Cbfb null allele (Cbfbrss/−) results in a 3-4 fold reduction in CBFβ protein levels. Although HSCs emerge in Cbfbrss/− animals, fetal T cell development is severely impaired. Here we examined the T cell developmental block in more detail by culturing fetal liver cells from Cbfbrss/− animals on OP9 stromal cells that express the Notch ligand Delta-like-1 (DL1) (Schmitt and Zúñiga-Pflücker, Immunity17: 749, 2002). Fetal livers (E14.5) from Cbfbrss/− animals contained normal numbers of both c-kit+Sca-1+lin- cells and c-kit+IL7r+ lymphoid progenitors. Lin- fetal liver progenitors cultured on OP9-DL1 cells in the presence of IL-7 and Flt3L displayed a growth disadvantage relative to wild type cells, and a block at the double negative 1 (DN1, CD44+ CD25−) stage of T cell development. The T cell defect could be rescued by retroviral transduction of the CBFβ heterodimerization domain into lin- fetal liver cells, but not by a G61A/N104A mutant that cannot bind the Runx subunits. Genes whose expression was decreased in DN1 cells purified from the OP9-DL1 cultures included CD3 and the early T cell transcription factors GATA3 and TCF. Although expression of several Notch pathway genes (Notch1, Hes-1/5, Deltex-1) was mildly decreased, Notch signals were clearly transduced, suggesting that Notch signaling was intact. These data demonstrate that reduced CBFβ levels impair the differentiation of stem cells/progenitors into T cells at the earliest stage of T cell development. This in vitro model will be useful for characterizing the molecular circuitry involving CBFβ in T cell development, and for identifying CBFβ protein partners.
Transcriptome Analysis of Murine Myeloid Development.
Authors: Yuan Wenlin , Ley Timothy J. 
Published: April 2020

Abstract
To characterize the transcriptional program that governs myeloid differentiation, we performed microarray analyses of cells derived from a G-CSF-dependent in vitro myeloid differentiation system. Using a high speed cell sorter, we purified CD34+/Lin− hematopoietic progenitors from wild type C57Bl/6 murine bone marrow cells cultured for 3 days in stem cell factor (SCF), thrombopoietin (TPO), interleukin-3 (IL3), and FLT3 ligand. These enriched progenitors were then treated for 7 days with G-CSF and SCF, which induces a coordinate wave of myeloid maturation. Linearly amplified cRNAs were prepared from cells harvested on each day of differentiation (d0-d7), labeled, and hybridized with Affymetrix Mouse 430v2.0 arrays. Comparing the percentage changes within two biological replicate sets by statistical analysis, we found that 51.5% of expressed probesets (15,693) were differentially regulated between day 0 and day 7. Among them, 10,627 probesets (34.9%) displayed expression levels higher than 500 (the mean expression level of all probesets was normalized to a value of 1500 before analysis) on at least one day. To avoid the high variability associated with probesets displaying low hybridization signals, we focused on these 10,627 probesets. The data showed the expected expression patterns for several genes that are known to be regulated during myeloid differentiation, including C/EBPa, Pu.1, c-Myb, CDP, c-Jun, M-CSF, GATA-1, AML1, CBFb, NE, MPO, MMP9, CD11b, lysozyme, and several others. In addition, the expression profiles were very consistent with the array data obtained using highly enriched human bone marrow-derived cells at early, mid, and late stages of myeloid development (Theilgaard-Monch et al., Blood105: 1785, 2005). However, our data also includes expression patterns from earlier stages of myeloid differentiation, which may be important to identify genes that are specifically expressed in committed myeloid progenitors. We organized the expressed genes into several categories, including transcription factors, cell cycle regulators, apoptosis genes, receptor and ligand in signal transduction pathways, etc. Some genes relevant for the pathogenesis of acute myelogenous leukemia (AML) also showed dynamic changes during normal myeloid differentiation, including (but not limited to) AML1, Flt3, HoxA9, Npm1, Ncoa2, Crebbp, Etv6, Meis1, Bcl2, Mcl-1, P53, Junb, Myc, and others. This dataset will be invaluable for evaluating the molecular events that underlie normal myeloid differentiation, and for defining the normal expression patterns of genes that are disregulated during the development of myeloid malignancies.
The Role of Alemtuzumab Compared to Antitymocyte Globulin in Non-Myeloablative Stem Cell Transplantation; Especially for the Prevention of GVHD and Immune Recovery.
Authors: Park In Hae , Cheong June-Won , Lee Jee Young , Kim Jin Seok , Chun Hyun Ju , Kim Yuri , Park Sun Young , Lee Jin Koo , Min Yoo-Hong 
Published: April 2020

Abstract
Alemtuzumab is an anti-CD52 antibody, and has been used for lymphoid malignancies or as a member of non-myeloablative conditioning regimen in allogeneic transplantation. Especially, in non-myeloablative stem cell transplantation (NST), it has been reported that alemtuzumab is effective for graft-versus-host disease (GVHD), but the immune reconstruction after transplantation is delayed. In this study, we comparatively evaluated the efficacy of alemtuzumab for non-myeloablative conditioning, GVHD prophylaxis, and immune recovery in NST for hematologic diseases. We have compared the results in 28 recipients of a sibling or unrelated NST enrolled. The recipients were divided into 2 groups according to the use of alemtuzumab. In group A (n=21), the conditioning regimen was a combination of fludarabine, cyclophosphamide (or busulfan) and antithymocyte globulin (ATG), and group B (n=7) received fludarabine, cyclophosphamide (or busulfan), and alemtuzumab instead of ATG. GVHD prophylaxis was by cyclosporin A or FK506 plus methotrexate. There were no significant differences in the graft engraftment and period of granulocyte colony-stimulating factor infusion. Patients receiving alemtuzumab had a significantly lower incidence of acute GVHD (stage 2 or more) (14.3% versus 38.1%, P=0.03) and chronic GVHD (14.3% versus 52.4%, P=0.005). The relapse rate after transplantation was 28.6% (6 patients) in group A and 14.3 (1 patients) in group B (P=0.04). Flow cytometric analysis of peripheral mononuclear cells for evaluation of immune recovery showed that T-cell and NK-cell recovery were delayed in both groups. However, T-cell and NK-cell recovery after transplantation occurred earlier in patients received alemtuzumab. No significant differences were observed in disease-free or overall survival between two groups. In conclusion, alemtuzumab can be recommended for immune suppression in NST, with successful control over acute/chronic GVHD and inducing relatively earlier immune recovery after transplantation.
Allogeneic Blood Stem Cell Transplantation in Patients with Hematologic Diseases after Conditioning with Treosulfan and Fludarabine.
Authors: Casper Jochen , Wilhelm Stefan , Steiner Beate , Wolff Daniel , Grobe Norbert , Haehling Detlev , Hartung Gernot , Hilgendorf Inken , Lueck Andreas , Junghanss Christian , Kahl Christoph , Pichlmeier Uwe , Freund Mathias 
Published: April 2020

Abstract
Background and objective:
Allogeneic transplantation can not be offered to many patients due to potential side-effects of conventional conditioning. Dose-reduced conditioning approaches improve tolerability, however, treatment intensity and efficacy might be reduced as well. We have, therefore, developed a dose intense, but toxicity reduced conditioning regimen based on treosulfan and fludarabine and report first results.
Patients and methods:
65 patients with a median age of 50 years were transplanted from related (n=21) or unrelated donors (n=44) after conditioning with treosulfan (3x10, 3x12 or 3x14g/m2 i.v.) and fludarabine (5x30mg/m2 i.v.). 21 patients were in complete remission (CR) and 44 patients had not reached a CR at the time of transplantation. 59 of 65 patients were considered unfit for a conventional conditioning regimen.
Results:
After a median follow up of 3.2 years an actuarial overall survival (OS) of 59.2 % (at 3 years) and an event-free survival (EFS) of 40,1% has been reached. Patients with related donor or transplantation in CR had a better OS (85,4 resp. 74,2%) and EFS (52,2% resp. 61,9%). The cumulative incidence of relapse at 3 years was 26,2%. Transplantation in CR resulted in a lower relapse incidence than transplantation not in CR (17,9 vs. 30%). Treatment-related mortality (TRM) at day 100 is 17,4% (cumulative incidence). In stepwise Cox regression analyses for OS, EFS and TRM only the covariables transplantation in CR vs. not in CR and the donor status were shown to be influential. No effects were detected for age (under 50 years vs. 50 years and older), concomitant disease (present vs. not present) or intensity of pretreatment.
The cumulative incidence for acute graft-versus-host disease (GvHD) °I-IV at day 100 was 43.3%. The cumulative incidence for chronic GvHD at 3 years is 36.1%.
Conclusions:
These results with a conditioning therapy of treosulfan and fludarabine indicate that patients despite higher age, concomitant disease or after intensive pretreatment can be successfully transplanted without increased treatment-related mortality.
Identification of Novel MHC Class I and Class II Epitopes of WT1 Using a Peptide Library Screen.
Authors: Rezvani Katayoun , Mielke Stephan , Kilical Yasemin , Grube Matthias , Fujiwara Hiroshi , Sconocchia Giuseppe , Lee Jongming , El-Ouriaghi Frank , Hensel Nancy , Barrett A. John 
Published: April 2020

Abstract
Although several HLA-A*0201-restricted immunodominant peptides from the leukemia-restricted protein WT-1 are characterized, T cell responses to peptide sequences binding to other common class I and II epitopes of WT-1 remain almost completely unexplored. A more comprehensive definition of the WT-1 antigen would extend peptide antigen vaccines to individuals lacking HLA-A*0201 and *2402 and improve vaccine potency by recruiting both CD4+ and CD8+ T cell responses. Here we used a WT1 peptide library to identify WT-1 peptide sequences inducing CD4+ and CD8+ T cell responses in normal individuals and patients with AML and other myeloid leukemias. Six cases were studied. The library consisted of 110 15mer peptides overlapping by 11aa covering the entire WT-1 protein in 21 pools. Monocytes were isolated by plastic adherence and pulsed with peptide pools for 3 hours. Autologous CD8+ and CD4+ T cells were then added. Pools of peptides were prepared in such a way that each peptide was represented in two different peptide pools, allowing the identification of the respective peptide by responses in the two corresponding pools. Cells were harvested for RNA extraction and reverse transcription. Real time PCR (RQ-PCR) was used to identify peptide-specific induction of IFN-γ and IL-2 in CD8+ and CD4+ T cells. The SYFPEITHI binding motif software was then used to predict the probable HLA restriction for the candidate epitopes. To confirm candidate peptide immunogenecity and HLA restriction, selected peptides were synthesized and tested individually. In addition to the known HLA-A*0201 peptides WT37, WT126, WT187 and WT235 we identified 20 new MHC class I and II epitopes of WT1. Four were restricted by more than one HLA allele, demonstrating the promiscuity of epitope binding. One epitope (VPGVAPTLV) was restricted to HLA-A*0201 and HLA-B*5101. One epitope (SGQFTGTAGACRYGP) was restricted by a class I HLA allele, namely HLA-*6801 and a class II HLA allele, DR*1501. Two epitopes (YGPFGPPPPSQASGQ and QKKFARSDELVRHHN) were restricted by multiple MHC class II alleles. The proliferative response of CD4+ and CD8+ T cells to candidate peptides was confirmed using CFSE labeling. We now plan to characterize the antileukemic effects of CD4+ and CD8+ T cells induced by these peptides with a view to designing broad-spectrum vaccines inducing leukemia-reactive T cells across a wide range of HLA types.