bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prevention of Graft-Versus-Host Disease in Mouse Model Using Anti-Mouse C5 Antibody.
Authors: Nishimura Jun-ichi , DeOliveira Divino , Chen Benny J. , Kanakura Yuzuru , Rother Russell P. , Chao Nelson J. 
Published: October 2019

Abstract
Graft versus-host disease (GVHD) is a major cause of morbidity and mortality after bone marrow transplantation (BMT). An allogeneic GVH reaction is a response of donor lymphoid cells to host minor or major histocompatibility antigens. Donor T cells can be activated through the innate and the adaptive immune mechanisms. Donor B cells produce antibodies directed to host cells. These mechanisms may activate complement pathways. Thus, complement may have a crucial role in inflammation during a GVH reaction, but direct evidence for this has not been shown. In this study, we investigated the possibility of complement inhibitor, anti-mouse C5 antibody (BB5.1), to ameliorate the symptoms of GVHD using an acute GVHD mouse model: C57BL/6 (H2b) →BALB/c (H2d). One million T cells were injected together with 1 x 107 T-cell-depleted bone marrow (TCD BM) cells via tail vein into lethally irradiated BALB/c (8.5 Gy) recipients. Anti-mouse C5 antibody or its isotype matched control was administered intraperitoneally at a dose of 1 mg/mouse, 3 doses/week, for 4 weeks. Recipients were weighed weekly, and their survival was monitored daily. Average body weight of C5 antibody treated mice was 15.8 g at day 84 (19.2 g at day 0, N=12), whereas average weight of control mice was 13.3 g (19.2 g at day 0, N=12) (P=0.05, Student’s t-test). Kaplan-Meier survival curves were also compared as shown in the Figure. Eight of 12 mice were alive at day 84 in the treated group, as compared to only 2 of 12 in the control group (P=0.03, Logrank test). A second experiment showed similar data. We, thus, observed the effect of anti-mouse C5 antibody to reduce the symptoms of GVHD using an acute GVHD mouse model. These results might open a new window for the prevention of acute GVHD. Further experiments are currently ongoing to clarify the exact mechanism between complement and GVHD.
Figure Figure
Damage to Aging Thymic Stroma and De Novo T Cell Development.
Authors: Prockop Susan , Beesen Ana , Myers Aaron , O’Reilly Richard J. 
Published: October 2019

Abstract
Successful T lymphopoiesis is essential for maintaining resistance to infection, mediating graft versus tumor activity, and as a platform for some immune based therapies. Among the most profound age-related change affecting the hemopoietic system is involution of the thymus and a decline in T cell production. Coordinated interaction of marrow-derived lymphoid progenitors with thymic stromal cells is required for successful de novo T lymphopoiesis. Current models of thymic involution indicate that aging alters the capacity of thymic stromal cells to support this de novo T cell lymphopoiesis. Loss of specific sets of thymic stromal cells, or loss of specific lympho-stromal signaling interactions may partially explain age-based impaired T cell reconstitution. Prior models examining the potential to reconstitute the aged thymus have been conducted either in vitro or in cyto-reduced hosts, and thus largely risk confounding cytotoxic damage with age-related damage. We have established an in vivo functional assessment of the ability of thymic stroma to support de novo T lymphocyte development. In the absence of radiation or chemotherapy this system supports robust thymic reconstitution after transplantation of limited numbers of T cell depleted wild type precursors. We have previously demonstrated that the capacity for reconstitution in this model system depends on the presence of normal, but empty, stromal niches. Using this model we have demonstrated that a variety of chemotherapeutic agents damage the thymic stroma and impair robust reconstitution. We now present data demonstrating that with increasing age reconstitution becomes less robust. Thus in mice older than 16 months unlike in those younger than twelve months, donor derived T lymphocytes can be identified, but the absolute cellularity of the thymus does not increase after transplant. In addition, we demonstrate that there are agent specific defects in the stromal compartments of treated aged mice. We have proceeded to identify a subset of agents (including split dose irradiation) that impair reconstitution in aged recipients but do not do so in the young. Thus while reconstitution of the thymus of young mice subjected to hyperfractionated total body irradiation declines 28 +/− 3% that of old mice treated in the same manner declines 91 +/− 6%. This last finding has enabled us to identify specific “at risk” stromal populations that normally appear to support the DN2 to DN3 transition in T lymphopoiesis. In a series of experiments designed to ask whether these at risk stromal populations can be protected from injury we identify agent specific differences in mechanisms of injury. It is anticipated that results from these investigations will augment our understanding of lympho-stromal interactions crucial to normal intra-thymic T cell development.
Ancestim (r-metHuSCF) Plus Filgrastim for the Mobilization and Collection of Peripheral Blood Progenitor Cells for Autologous Transplant after Failure of Prior Mobilization: The French Experience of Compassionate Use.
Authors: Lapierre Valerie , Chabannon Christian , Makowski Caroline , Dumesnil Cecile , Valteau-Couanet Dominique , Vekhoff Anne , Caillot Denis , Azar Nabih , Divine Marine , Blakolmer Karin , Cailliot Christian , Moreau Philippe , Rossi Jean-Francois 
Published: October 2019

Abstract
Background: Ancestim (r-met human stem cell factor; Amgen Inc, CA, USA) is available in France for compassionate use in patients who have failed prior mobilization or an insufficient number of CD34+ cells for autograft. We retrospectively reviewed data from these patients to evaluate the efficacy of ancestim combined with filgrastim in achieving an adequate stem cell yield.
Patients and methods: Between January 1998 and July 2007, permission for ancestim use was granted by the French health authority and Amgen to 550 patients (pts) at 82 sites (median age 55 yrs (range, 18–74); 50% male). Non-Hodgkin’s lymphoma (NHL) accounted for 50% of pts, multiple myeloma (MM) 28%, chronic lymphocytic leukaemia 6%, Ewing’s sarcoma 4%, neuroblastomas 3%, ovarian carcinoma 1%, osteosarcomas 1%, and other tumours 7%. Of the 508 pts for whom data were available and analysed, 378 had undergone prior mobilization and 117 were considered to have insufficient harvests; mobilization history was unknown for 13 pts. Fifty-eight percent of the pts were treated with ancestim (20 microg/kg/day s.c.) and filgrastim (10 microg/kg/day s.c.) alone for a median of 6 days (range, 2–21), while the other pts received chemotherapy followed by ancestim (20 microg/kg/day s.c.) and filgrastim (5 microg/kg/day s.c.) for a median of 11 days (range, 1–33).
Results: The target threshold of 2x106 CD34+ cells/kg was reached in 198/508 pts (39%) (including 51% of the 144 MM pts and 31% of the 254 NHL pts) after a median of 2 leukaphereses (range, 1–6). 35% of pts with prior mobilization failure and 52% of pts with prior insufficient harvests reached the target of 2 x106 CD34+ cells/kg, with a median number of CD34+ cells harvested of 3.4 x106/kg (range, 0.5–39.5). Of these, 150 pts (76%) proceeded to autologous transplantation. Data on engraftment are available for 137 pts: the median times to reach 0.5 x 109/L neutrophils and 20 x 109/L platelets were 12 days (range, 6–40) and 13 days (range, 0–31), respectively. All pts receiving ancestim were premedicated and treatment was generally well tolerated. Four pts were reported to have experienced severe mast cell-mediated reactions, none of which were life-threatening.
Conclusion: Ancestim plus filgrastim, alone or with chemotherapy, successfully rescued adequate CD34+ cell mobilization and collection in preparation for autologous transplantation in approximately 40% of pts.
Cryopreservation of Peripheral Blood Stem Cells (PBSC) with Hydroxyethylstarch (HES) and Dimethylsulfoxide (DMSO) Results in Faster Granulocyte Recovery Than Using Dimethylsulfoxide Alone.
Authors: Chan Wai Shun , Ma Michael , Bhatty Muhammad U.A. , Preti Robert , DeVito Marie , Pecora Andrew L. , Donato Michele L. , Feldman Tatyana , Goldberg Stuart L. , Goy Andre , Siegel David S. , Rowley Scott D. 
Published: October 2019

Abstract
PBSC components are routinely frozen for storage before autologous transplantation in the treatment of malignant diseases. We have used 5% DMSO, 3.75% human serum albumin (HSA), 0.2% dextrose with 6% HES for cryopreservation of cells (DMSO/HES), but the lack of a commercial source of HES required us to evaluate other cryopreservative solutions. A solution containing 5% DMSO, 2.25% HSA in normal saline (DMSO) was selected from a range of cryoprotectant solutions based on in vitro data demonstrating equivalence in preservation of hematopoietic colony forming ability to the HES solution previously used at this center. Fifteen months after the change, we performed a retrospective review of engraftment kinetics in autologous PBSC transplantation for the treatment of patients classified by cryoprotectant solution used. PBSC collected before 13-June-2006 were frozen in the DMSO/HES solution. PBSC collected after that date were frozen in DMSO. Patients underwent transplantation using dose-intense preparative regimens appropriate to diagnosis. Filgrastim was administered after transplantation to hasten granulocyte recovery. All patients after the change date were evaluated and patients treated in a similar time frame before the change were selected for comparison. Patient demographics and mean CD34+ cell number infused were similar for these two groups.
Demographics N Age CD34+ Cells/Kg (xE6) Gender (M:F) Diagnosis (MM:NHL:Other) Shown are median age and mean CD34+ cells/kg transplanted. Diagnosis is MM=multiple myeloma, NHL=non-Hodgkin’s lymphoma, and Other = other hematological malignancies. DMSO/HES 122 56 7.6 59:63 75:34:13 DMSO 103 58 7.1 55:48 79:14:10 p Value 0.26 0.56 0.17 0.84
Patients who received PBSC frozen in DMSO/HES achieved a significant one-day faster granulocyte recovery but similar platelet recovery compared to patients receiving PBSC frozen in DMSO.
Engraftment Kinetics N ANC>500 ANC>1000 Platelet>20,000 Shown are median days to achieve particular endpoint of engraftment. DMSO/HES 122 11 12 13 DMSO 103 12 13 14 p Value <0.001 0.002 0.83
Despite similar in vitro and patient demographic profiles, these data indicate that the combination cryoprotectant solution results in a clinically significant enhancement in speed of engraftment, resulting in a one (1) day reduction in hospital stay associated with the transplant regimen. The effect appears to be lineage-specific. Other modifications of cryoprotectant solutions may achieve yet faster engraftment, benefiting patients undergoing autologous PBSC transplantation by shortening the period of marrow hypoplasia. These results agree with another study of cryopreservation with and without HES (Rowley, et al. Bone Marrow Transpl2003;31:1043–51).
Molecular Studies of Neutropenia in Barth Syndrome.
Authors: Aprikyan Andrew A. , Makaryan Vahagn , Dale David C. 
Published: October 2019

Abstract
The Barth syndrome is a rare mitochondrial disease that causes dilated cardiomyopathy, skeletal myopathy, and severe neutropenia. It has been reported that at least two patients with Barth syndrome died from septicemia attributed to neutropenia. The disease is an X-linked autosomal recessive disorder caused by mutations in the G4.5 or TAZ (tafazzin) gene, which introduce truncations, substitution, or alternative splice sites. Hematological investigations revealed normal chemotaxis, but the presence cytoplasmic vacuoles in myeloid cells and maturation arrest of neutrophil development at the myelocyte stage. Thus, a defect in neutrophil formation appears to be the primary reason for neutropenia and the susceptibility to infections in the Barth syndrome patients. Recent studies were mostly focused on cardiac, genetic and metabolic abnormalities associated with the disorder. However, the neutropenia aspect of the Barth syndrome remains unclear. The TAZ gene mutations appear to truncate the tafazzin protein likely resulting in the loss of function. Although a drosophila model of the Barth syndrome was recently reported, the cellular or mouse models of this disorder are not available yet. Thus, the link between TAZ mutations and severe neutropenia remains unknown. We hypothesized that TAZ mutations, which lead to the loss of function of tafazzin protein, trigger impaired cell survival and reduced production of neutrophils and their neutrophil precursors, thus resulting in severe neutropenia in Barth syndrome. To test this hypothesis, we attempted to knock-down the expression of the tafazzin gene in human myeloid progenitor cells using TAZ-specific shRNA. Four shRNA sequences specific to exons 4 through 7 were used for transfection of human myeloid progenitor cells that were later examined by flow cytometry and Western blot analyses. At least 2 of the shRNA constructs resulted in a substantial down-regulation in the expression level of the tafazzin protein in transfected myeloid progenitor cells as determined by Western blot. Flow cytometry analyses revealed that the knock-down of TAZ gene expression was associated with approximately 40–50% increase in proportion of apoptotic annexin-positive cells compared with cells transfected with control scrambled shRNA. These data suggest that the loss of function of TAZ gene is cytotoxic to hematopoietic cells and that severe neutropenia is due to the accelerated apoptosis of myeloid progenitor cells in patients with Barth syndrome. Further studies needed to elucidate the specific signaling pathways and to identify potentially therapeutic agents capable of controlling accelerated apoptosis of myeloid cells in Barth syndrome.
Predictors of Transformation to Myelodysplasia/Acute Myelogenous Leukemia (MDS/AML) in Severe Congenital Neutropenia (SCN).
Authors: Boxer Laurence A. , Bolyard Audrey Anna , Newburger Peter E. , Bonilla Mary Ann , Kannourakis George , Dror Yigal , Link Daniel C. , Alter Blanche P. , Rosenberg Philip S. , Dale David C. 
Published: October 2019

Abstract
Severe congenital neutropenia (SCN) is a heterogeneous disorder of myelopoiesis characterized by an absolute neutrophil count (ANC) persistently below 0.50 x 109/L (500/[um]L), with maturation arrest of neutrophil precursors of the bone marrow at the promyelocyte/myelocyte stage. G-CSF treated and untreated SCN patients are at risk of developing MDS/AML. Clinicians caring for these patients must vigilantly observe for evidence of evolution to malignancy and continually consider hematopoietic transplantation as an alternative therapy. We have previously reported that SCN patients requiring higher daily doses of G-CSF treatment (> 8 mcg/kg/day) are at an increased risk of MDS/AML, and that this risk is not predicted by pre-treatment bone marrow examinations or the pre-treatment ANC. We have reviewed clinical data for 46 patients over a 15-year period who were referred to the Severe Chronic Neutropenia International Registry (SCNIR) and developed MDS/AML. Four patients referred with a history of SCN had increased blasts on the bone marrow evaluation prior to G-CSF treatment or enrollment in the SCNIR. Blood counts were not available for two patients. For the other 40 patients, all treated with G-CSF for a median of 79 months (range 0.8 to 182 months), comparison of blood counts for the periods 12–24 months and 0–3 months before the diagnosis of MDS/AML showed:

Changes in blood counts for the patient population may suggest a potential for malignant transformation. In these 40 cases, the principal clinical findings leading to the diagnosis of MDS/AML were: routine bone marrow surveillance 38%, decreased peripheral blood counts 26%, increased infections 10%, increased blasts in the blood 5%, decreased ANC with requirement for increased G-CSF dosage 2%, hepatosplenomegaly 2%, other (1 septal panniculitis and 1 leukemia cutis) 5%, and unknown 12%. The finding of mutations in the ELA2 gene in 10 of 15 cases (67%) did not influence the likelihood of MDS/AML. Previously we found that the duration and dose of G-CSF and alterations of the ANC on G-CSF therapy, suggest a high risk of transformation over time. Based on these data, the SCNIR recommends that patients with SCN have regular surveillance of blood counts every 1 to 2 months and annual bone marrow examinations with cytogenetic studies. Our findings suggest that a decline or change in blood counts of individual patients over a 3-month period may be informative in recognizing transformation to MDS/AML.
Cost-Effectiveness Analysis in Canada of Dasatinib Versus Imatinib for the Treatment of Chronic Myelogenous Leukemia in Patients with Imatinib Resistance.
Authors: Sambrook Jennifer C. , Zou Denise , Taylor Matthew J. , Davis Catherine C. , Joyeux Alexandre , Levy Adrian R. 
Published: October 2019

Abstract
Objective: To estimate the cost-utility of dasatinib versus high-dose imatinib (HDI, imatinib 800mg/day) for chronic myelogenous leukemia (CML) in patients with resistance to imatinib at standard doses.
Methods: We adapted to the Canadian setting a Markov model with a monthly cycle length that followed hypothetical patients until death. Cost effectiveness was evaluated during three phases of CML: chronic (86% of Ontario patients in 2006), accelerated (9%) and blast crisis (5%). Efficacy of dasatinib and HDI (cytogenetic and hematologic response) and adverse events (AEs) were taken from randomized trials. Canadian costs of medications, laboratory tests, professional fees, hospitalizations, and AEs were obtained from published sources. Societal utility scores were estimated from a Canadian time trade-off study. Analyses were performed from the perspective of the Ontario Ministry of Health. Costs (2006 CDN$) and outcomes (QALYs) were discounted at 5% annually after the first year. Univariate and probabilistic sensitivity analyses were carried out to quantify uncertainty for the incremental cost-effectiveness ratios (ICERs).
Results: Dasatinib dominated HDI in chronic phase patients with additional QALYs (3.92 vs. 3.47) and a lower lifetime cost ($379,678 vs. $444,934). The ICERs of dasatinib vs. HDI in accelerated and blast phase were $88,098/QALY and $173,922/QALY respectively. The higher cost/QALY of dasatinib in the advanced phases is explained primarily by the increased predicted survival of dasatinib patients and therefore the longer duration of drug therapy for these patients. Parameters with the greatest influence on results were the time horizon, drug costs and utility values. The interpretation of results remained robust in univariate and probabilistic sensitivity analyses.
Conclusion: From an economic perspective dasatinib is an attractive treatment choice for the majority of imatinib resistant CML patients and provides better value for money than HDI.
Multisite Phosphorylation of NIPA at G2/M.
Authors: Illert Anna L. , Bassermann Florian , von Klitzing Christine , Seipel Petra , Morris Stephan W. , Peschel Christian , Duyster Justus 
Published: October 2019

Abstract
The regulated oscillation of protein expression is an essential mechanism of cell cycle control. The SCF class of E3 ubiquitin ligases is involved in this process by targeting cell cycle regulatory proteins for degradation by the proteasome, with the F-Box subunit of the SCF specifically recruiting a given substrate to the SCF core. We previously reported the cloning of NIPA (Nuclear Interaction Partner of ALK) in complex with constitutively active oncogenic fusions of ALK, which contributes to the development of lymphomas and sarcomas. Subsequently we characterized NIPA as a F-Box protein (FBP) that defines an oscillating ubiquitin E3 ligase. The SCFNIPA complex targets nuclear cyclin B1 for ubiquitination in interphase while phosphorylation of NIPA in late G2 phase and mitosis inactivates the complex to allow for accumulation of cyclin B1. Here, we identify the region of NIPA that mediates binding to its substrate cyclin B1. In addition to the recently described serine residue 354, we specify 2 new residues, Ser-359 and Ser-395, implicated in the phosphorylation process at G2M within this region. Moreover, we found cyclin B1/Cdk1 to phosphorylate NIPA at Ser-395 in mitosis. Mutation of both Ser-359 and Ser-395 impaired effective inactivation of the SCFNIPA complex, resulting in reduced levels of mitotic cyclin B1. Furthermore, we aimed to identify the kinases involved in the initial phosphorylation of Ser-345. Therefore, we tested a panel of different kinases active at the G2M transition such as GSK3?, Casein kinase 2, PLK-1 and Erk1. Effective in vitro phosphorylation of NIPA could only be demonstrated with Erk-1. Moreover, we demonstrate an interaction of Erk-1 and NIPA at G2M but not in interphase cells. Binding of Erk-1 and NIPA led to phosphorylation at Ser-354 in vivo and could be blocked by the MEK-1/MEK-2 inhibitor PD98059. Together these data suggest a process of sequential phosphorylation where NIPA is initially phosphorylated by Erk-1 leading to the dissociation of NIPA from the SCF core complex. Once Ser-354 is phosphorylated, cyclin B1/CDK1 amplifies phosphorylation of NIPA, thus contributing to the regulation of its own abundance in early mitosis. In ALK positive lymphomas enhanced phosphorylation of NIPA at Ser 354 can be observed. We demonstrate that NPM-ALK leads to the activation of Erk-1, thereby phosphorylating and inactivating the SCFNIPA E3 ligase. Inactivation of SCFNIPA may have an important impact on the cell cycle turnover of lymphoma cells and thus for the pathogenesis of NPM-ALK induced lymphomas.