bluuebunny/crossref_metadata_2025_split huggingface dataset:
Number of Prior Chemotherapy Regimens: Important Prognositic Factor of All-Cause Mortality after Allogeneic Hematopoietic Cell Transplantation.
Authors: Kiyokawa Tetsuyuki , Nagakura Shoichi , Hidaka Michihiro , Uike Naokuni , Hamaguchi Motohiro , Horibe Keizo , Sato Kensaku , Kawano Humio 
Published: October 2019

Abstract
BACKGROUND: Despite recent advances, mortality rates after allogeneic hematopoietic cell transplantation remain high and cannot accurately predicted.
PURPOSE: To determine the value of pretransplant factors in predicting all-cause mortality during first 1 year after allogeneic HCT.
PATIENTS AND METHODS: A retrospective review of 303 consecutive allogeneic hematopoietic cell transplants was performed with attention to mortality and pretransplant factors in three hematopoietic cell transplantation centers between 2000 and 2005 in Japan. Patients underwent transplantation from sibling (n=200) or unrelated donors (n=104) with myeloablative (n=188) or nonmyeloablative conditioning (n=116). Pretransplant factors and the hematopoietic cell transplantation-specific comorbidity index (HCT-CI: Sorror et. al.) were assessed with general linear model (GML) analysis.
RESULTS: Mortality before 1 year occurred in 145 of 303(48.3%) transplant recipients. The number of prior chemotherapy regimens and the HCT-CI were found to be useful predictors of early mortality. Kaplan-Meier estimates of survival as stratified by the HCT-CI (Fig.1) and the number of prior chemotherapy (Fig.2). The HCT-CI were useful for predicting outcomes. However, the sample size of patients with scores of 1 or more, captured by the HCT-CI did not exceed 15%. Futher, the number of prior chemotherapy regimens showed better suvival prediction.
CONCLUSION: The number of prior chemotherapy regimens will be a promising tool to identify patient at risk for all-cause mortality during first 1 year after allogeneic HCT.
Figure Figure
A Cellular Proteome Map of Human Multiple Myeloma.
Authors: Drach Johannes , Slany Astrid , Sagaster Verena , Gundacker Nina , Haudek Verena , Odelga Viktoria , Wimmer Helge , Zielinski Christoph , Gerner Christopher 
Published: October 2019

Abstract
Background and Aims: Molecular profiling identifies proteins characteristically deregulated in malignant diseases. Characteristic biomarkers may be useful to support diagnosis and patient stratification, while the recognition of aberrant cell activities and cell survival strategies may lead to the development of specifically designed pharmacologic strategies. We therefore performed proteomic profiling of primary human multiple myeloma (MM) cells in order to define the impact of protein expression abnormalities in MM.
Methods: Plasma cells were isolated from bone marrow of patients with MM or MGUS and forwarded to proteome analysis based on 2D gel electrophoresis in addition to shotgun analysis by nano-LC-MS/MS. Erythrocytes, platelets and plasma as well as quiescent and activated lymphocytes, monocytes, endothelial and dendritic cells from healthy donors were processed in an identical manner for comparative analysis. The resulting data were interpreted with the aid of a homemade SQL database.
Results: Among about thousand proteins identified in MM cells, we found aberrant expression of proteins involved in fatty acid beta-oxidation (VLCAD), unfolded protein response/ER-stress (ARMET protein, cytosol aminopeptidase), oxidative stress (ste20/oxidant stress responsive kinase 1), interferon response (MX1), iron uptake (transferrin receptor/CD71), DNA modification (transforming protein ERG, methyltransferase-like protein 7A), apoptosis and survival (apoptosis-inducing factor 1, hsp75), protein synthesis (ribosome-binding protein, Unc-13 D), cell adhesion (CD9/tetraspanin-29, LYRIC/metastatic adhesion protein), signaling (sts-1) and cell-cell interaction (Cystatin F, basigin/collagenase stimulatory factor, stem cell growth factor, small inducible cytokine B7, PD-ECGF/Gliostatin).
Conclusion: The presently applied proteome analysis strategy, based on the systematic investigation of purified primary human cells, allowed us to find characteristic alterations in MM cells. Several proteins directly relate to known aberrant cell activities such as elevated protein synthesis and secretion resulting in ER-stress, which makes the cells sensitive to proteasome inhibitor treatment. Work is in progress to use quantitative assessment of such proteins for patient stratification, identification of response predictors, and biomarkers for the distinction of MM and MGUS.
Overcoming Graft Rejection in Adult Patients with High-Risk Severe Aplastic Anemia Using Supplemental CD34+-Enriched PBSC in Combination with BM.
Authors: Cho Byung-Sik , Kim Sung-Yong , Kim Yoo-Jin , Eom Ki-Seong , Kim Hee-Je , Lee Seok , Min Chang-Ki , Kim Dong-Wook , Lee Jong-Wook , Min Woo-Sung , Kim Chun-Choo 
Published: October 2019

Abstract
The transplantation of a large number of stem cells can overcome the graft rejection with the pitfall of an increased incidence of graft-versus host disease (GVHD). We analyzed the clinical outcome of adult patients with severe aplastic anemia (SAA) at high risk for graft rejection who received a transplant with both bone marrow (BM) and CD34+-enriched peripheral blood stem cells (PBSC) to decrease the graft rejection. Between December 1999 and April 2006, 31 consecutive adult patients with SAA who were multi-transfused (more than 40 units of red blood cells ± platelets) and/or had a 3 years or longer period prior to transplant were investigated. The median age of patients was 39 years (range, 21–52) and 19 (61.3%) were women. The median interval from diagnosis to transplant was 62 months (range, 2–347). The median number of transfusions before transplant was 144 units (range, 20–680). Twenty-three (74.2%) patients received a prior immunosuppressive treatment and 3 (9.7%) prior allogeneic transplants. All patients received both BM and CD34+-enriched PBSC from sibling donors. PBSC were collected on days 5 and 6 of G-CSF (10μg/kg/day), and T cells were depleted using magnetic cell sorting (CliniMACS system, Miltenyi Biotec, Germany). Marrow harvesting was performed 48 hours after the last leukapheresis. Conditioning regimens consisted of cyclophosphamide/antithymocyte globulin + procarbazine (n=19) or fludarbine (n=9). Other 3 patients with a history of prior transplant received irradiatin based conditining regimens. GVHD prophylaxis consisted of cyclosporin and short-term methotrexate. Patients were transplanted with a median mononuclear cell dose of 1.35 (range, 0.43–4.05)×108/kg of body weight. The median CD34+ cell counts in the CD34+-enriched PBSC and unpurged BM were 7.99 (range 2.26–23.49)×106/kg and 4.41 (range, 1.34–15)×107/kg, respectively. With a median follow-up of 66 months, the 5-year probability of survival was 87.1%. The median time to neutrophils ≥0.5×109/L and platelet ≥20×109/L without platelet transfusions were 13 days (range, 10–32) and 16 days (range, 11–40), respectively. No patient developed graft failure. The cumulative incidence of acute GVHD over grade II and chronic GVHD were 9.7% and 16.1%, respectively. In conclusion, high rates of durable engraftment and excellent long-term survival were achieved by our approach which seems to play a role in overcoming the sensitization to histocompatibility antigens. Remarkably, the T-cell depleted PBSC apparently did not increase the incidence of GVHD. The high dose stem cell component therapy may be feasible for high-risk SAA adult patients.
Phenotypic and Functional Features of Vγ9/Vδ2 T Cells in Chronic Lymphocytic Leukemia Are Strongly Correlated with the Mutational Status of the Immunoglobulin Heavy Chain Variable Region (IgVH) and Metabolic Activity of the Mevalonate Pathway in Tumor Cells.
Authors: Coscia Marta , Peola Silvia , Matta Giorgia , Pantaleoni Francesca , Foglietta Myriam , Angelini Daniela , Chiaretti Sabina , Riganti Chiara , Guarini Anna , Drandi Daniela , Ladetto Marco , Bosia Amalia , Foà Robin , Battistini Luca , Boccadoro Mario , Fournie Jean Jacques , Massaia Massimo 
Published: October 2019

Abstract
Vγ9/Vδ2 (γδ) T cells have an important effector and regulatory role in innate and adaptive immunity against microbes, stressed cells and tumor cells. They represent less than 5% of peripheral blood lymphocytes, but can be activated and expanded in vitro by aminobisphosphonates (ABP) like zoledronic acid (Zol), as surrogates of their natural ligands. In this study, we have investigated the proliferative response of γδ T cells to Zol in 59 patients with chronic lymphocytic leukemia (CLL) at diagnosis. Proliferation of γδ T cells was observed in 30 patients (51%)(responders, R), whereas 29 patients (49%) were non-responders (NR). γδ T-cell subset distribution [e.g., naive (TN), central memory (TCM), effector memory (TEM), and terminally differentiated effector memory (TEMRA)] was well balanced in R patients, whereas TEM and TEMRA were largely predominant in NR patients. TEMRA of NR patients had a unique profile of NK receptors expression, characterized by the expression of the ILT4 inhibitory receptor, whereas R patients tended to have an higher expression of the costimulatory molecule NKG2D. Notably, HLA-G, a negative prognosticator in CLL and a well known ILT4 ligand, was more expressed in tumor cells of NR patients. This correlation prompted us to determine whether other tumor cell features were different in R and NR patients. Indeed, the latter showed significantly higher tumor cell counts, higher frequency of poor-risk cytogenetic abnormalities, and a significantly shorter median time to first treatment. Lastly, a very significant association was observed with the mutational status of the tumor immunoglobulin heavy chain variable region (IgVH), which was mutated (M) in 96% of R patients, and unmutated (UM) in 74% of NR patients. To gain further insight into this association, we evaluated the activity of the mevalonate pathway in tumor cells of M and UM patients. This pathway, that can be targeted by Zol, generates the phosphoantigens naturally recognized by γδ T cells. Bioinformatic and biochemical approaches showed that this pathway is more active in tumor cells of UM than M patients. Based on these data, we propose that tumor cells of UM CLL patients can more easily engage γδ T cells and, in the long run, exhaust their immune surveillance potential by driving their differentiation into functionally impaired TEMRA. In conclusion, we have identified a novel mechanism of immune escape which can contribute to the disease progression in UM CLL patients.
Tumor Associated Macrophages (TAM) in Skeletal Plasmacytomas of Patients with Multiple Myeloma.
Authors: Russell Stephen J. , Dogan Ahmet , Ong Hooi Tin , Vrana Julie , Dispenzieri Angela , Dietz Allan B. , Peng Kah-Whye 
Published: October 2019

Abstract
In multiple myeloma, the neoplastic plasma cells are either dispersed throughout the red marrow spaces of the axial and proximal appendicular skeleton or concentrated in discrete, well vascularlized plasmacytomas that stimulate osteoclastic bone resorption and erode into cortical bone. Interactions between bone marrow stromal cells (BMSC) and myeloma cells are clearly important in the pathogenesis of myeloma that diffusely infiltrates the bone marrow and where plasmacytomas are in contact with the bone marrow space. However, these interactions cannot be of major relevance at the centers of expanding plasmacytomas since these sites are devoid of osteoclasts, osteoblasts and other BMSC. Histological sections of skeletal plasmacytomas from 22 patients with multiple myeloma were therefore examined after immunostaining with anti-macrophage antibodies. Hematoxylin and eosin stained sections showed sheets of plasma cells interspersed with numerous small blood vessels. Immunostained sections revealed abundant CD68+, CD163+ macrophage infiltrates in all tumors, and scattered collections of CD3+ T lymphocytes. Tumor-associated macrophages (TAM) accounted for 2 to 12 percent of nucleated cells in these plasmacytomas and were evenly distributed through the parenchyma or clustered around CD34+ blood vessels. In general the TAM had dendritic morphology, and each dendrite was in close contact with multiple plasma cells. In a small percentage of cases, there was a subpopulation of TAM strikingly clustered around CD34+ positive blood vessels. However, vascular mimicry was not seen. Abundant murine CD68+ TAM were also found in subcutaneous KAS6/1 and MM1 myeloma xenografts in SCID mice. To determine whether cells of the monocytic lineage might be exploitable as carriers for systemic delivery of therapeutic agents to plasmacytomas, monocyte-derived dendritic cells were infected with oncolytic measles viruses and administered intravenously to SCID mice bearing subcutaneous KAS6/1 xenografts. Immunostaining for human MHC II antigen revealed that the infused DCs could localize to the KAS6/1 tumors where they heterofused with and transferred MV infection to the myeloma cells. This is the first report of TAM in skeletal plasmacytomas of myeloma patients. Further research into the role of TAM in supporting myeloma growth is warranted since they may offer many potential opportunities for the development of novel approaches to myeloma therapy.
Regulation of Chronic Lymphocytic Leukemia Cell Proliferation by CTLA4.
Authors: Mittal Amit K. , Rohlfsen Rae A. , Joshi Avadhut D. , Hegde Ganapati V. , Bociek R. Gregory , Joshi Shantaram S. 
Published: October 2019

Abstract
B-cell chronic lymphocytic leukemia (CLL) is the most common adult leukemia with heterogenous clinical course. Prognostic markers such as CD38 expression has proven to be reliable in predicting clinical outcome where high CD38 expression correlates with poor prognosis and low CD38 expression is associated with good prognosis. Recently, our lab has shown that expression of CTLA4 in CLL cells is inversely correlated with CD38 expression using microarray profiling and real time PCR. Microarray analysis showed significant differential expression of CTLA4 between high and low CD38 subgroups (n=49) (Joshi et al. Clinical Cancer Research, 2007, in press). However, whether CTLA4 plays a direct role in CLL pathogenesis or not is unknown. Therefore to study the potential role of CTLA4 in CLL, CTLA4 expression was down regulated in primary CLL cells from six different patients using antisense oligonucleotide. The down regulation of CTLA4 expression was confirmed by RT-PCR analysis. The proliferative capability of the CTLA4-down regulated CLL cells was determined using MTT assay as well as radiolabeled thymidine uptake assay at 24, 48 and 72 hours in culture. There was a significant increase in the proliferation of CTLA4-down regulated CLL cells compared to untreated control CLL cells or CLL cells treated with an irrelevant oligonucoeotide. To further understand the CTLA4 mediated regulation of CLL cell proliferation, expression levels of downstream target molecules of CTLA4 such as NFATC2 and c-FOS was studied using RT-PCR. There was a significantly increased expression of target genes NFATC2 and c-FOS, molecules in the CTLA4-downregulated CLL cells compared to control CLL cells. In addition, further analyses of the microarray data revealed that CD38 high expressing CLL cells (n=22) has significant (p<0.05) higher NFATC2 and c-FOS expression compared to CD38 low expressing CLL cells (n=29). This was further confirmed by real time PCR analyses (figure1). Thus, these results clearly demonstrate that CTLA4 is involved in the regulation of proliferation of CLL cells via regulating the expression of NFATC2 and c-FOS, the downstream signaling molecules involved in the cell proliferation. (This work was supported by the CLL Foundation, Houston, TX).
Figure Figure
Osteopontin in Hemophilic Synovitis.
Authors: Hakobyan Narine , Kaushal Megha , Enockson Candace , Valentino Leonard A. 
Published: October 2019

Abstract
Introduction Hemophilia is a congenital bleeding disorder due to the deficiency of blood coagulation factor VIII or IX. The joint is the most common site of serious bleeding. Repeated episodes of hemarthrosis lead to hemophilic synovitis (HS) and destructive hemophilic arthropathy (HA). The molecular changes responsible for HS and HA are not known. Osteopontin (OPN) has been implicated in the process that leads to bone destruction in other forms of arthritis. We hypothesized that OPN plays a similar role in HA.
Methods Hemophilic synovitis was induced in factor (F) VIII and IX deficient mice by causing traumatic (blunt trauma) and non-traumatic (needle puncture) hemarthrosis in the right knee joint of mice. The concentration of OPN was measured in plasma of mice five days after inducing hemorrhage. Expression of OPN in synovial tissue from affected knee joints was compared with control uninjured joints by DNA microarray and semi-quantitative RT-PCR analysis on different days following joint injury.
Results OPN concentration in the plasma of both FVIII and FIX deficient mice sacrificed 5 days after a single capsular puncture was increased 92–98% compared to control mice. In other experiments, traumatic hemarthrosis was induced three times at weekly intervals in the right knee joints of FVIII deficient mice. On day 21, synovial tissue from the knee joints of 20 mice was isolated, pooled and RNA from affected and control joints probed using microarray. The expression of OPN in right knee tissue was 7.5-fold greater compared to the left. These data were confirmed by RT-PCR analysis which showed that OPN was highly expressed in synovial tissue from mice 14, 21 and 30 days after single puncture injury and 21 days following three traumatic injuries.
Conclusion The concentration of OPN was increased in the plasma of mice following hemarthrosis and OPN expression was increased 7-fold in synovial tissue of mice with hemophilic arthropathy. Our data implicate OPN in the pathobiology of hemophilic arthropathy.
Figure Figure Figure Figure
Experience with a Modified Nijmegen-Bethesda Method for Measurement of Inhibitors in Hemophilia Patients: The CDC Inhibitor Surveillance Pilot Project.
Authors: Miller Connie H. , Platt S. Jean , Abshire Thomas , Brettler Doreen , Brockenstedt Paula , DiPaola Jorge , Massey Gita , Neff Anne , Shapiro Amy , Tarantino Michael , Wicklund Brian , Creary Melissa , Soucie J. Michael 
Published: October 2019

Abstract
A pilot project of a prospective surveillance system for coagulation factors VIII and IX inhibitors has been initiated at 9 U.S. Hemophilia Treatment Centers (HTCs). More than 500 patients have been enrolled. Risk factor and product exposure data are recorded at each HTC. A blood specimen is collected upon entry, annually, at product switch, or for clinical indication and tested centrally at the CDC, using a modified Nijmegen-Bethesda method. Plasma from specimens collected in 4.5mL evacuated tubes containing 3.2% sodium citrate is shipped overnight on cold packs. Specimens are heated to 56oC for 30 minutes to remove endogenous and infused FVIII and centrifuged. Specimens are initially screened for inhibitor using a single dilution of 3 parts patient plasma to 1 part normal pool plasma buffered with imidazole to pH 7.4 (BNPP). Specimens showing inhibition and those from previously positive patients are tested in multiple dilutions at 1 part patient dilution to 1 part BNPP. Dilution is in naturally FVIII-deficient plasma containing normal von Willebrand factor. After a 2-hour incubation at 37oC, FVIII remaining in the patient mixture is divided by FVIII remaining in a 1:1 mixture of BNPP and FVIII-deficient plasma and expressed as % residual activity (RA). %RA is converted to Nijmegen-Bethesda units (NBU) using a curve with one NBU equal to 50% RA. An inhibitor plasma of known titer is run with each assay as positive control. Split specimens shipped frozen and on cold packs showed a correlation of 0.998. 3:1 and 1:1 mixtures showed a correlation of 0.97. Almost 50% of the first 200 specimens received had measurable FVIII activity. The heating step was introduced to remove FVIII without damaging the antibody. 65 specimens went from >100% RA to a titer of 0–0.2 NBU after heating. Among 538 specimens tested for FVIII inhibitors, 435 (81%) were from patients with no previous history of inhibitor (shown below). 429 (98.6%) were < 0.5 NBU. The 6 specimens (1.4%) with > 0.6 NBU are under investigation as possible seroconversions.
Nijmegen-Bethesda Units in Patients with No Previous Inhibitor NBU No. Pts. (%) NBU No. Pts. (%) NBU No. Pts. (%) 0 220 (50.6%) 0.4 3 (0.7%) 0.8 1 (0.2%) 0.1 127 (29.2%) 0.5 0 0.9 0 0.2 59 (13.6%) 0.6 0 1.0–4.0 2 (0.5%) 0.3 20 (4.6%) 0.7 2 (0.5%) 5.0–19.0 1 (0.2%)
Among 121 specimens tested for factor IX (FIX) inhibitors, 113 were from patients with no previous history of inhibitor and all had NBU <0.2, using a comparable FIX inhibitor assay. Testing in a central laboratory by uniform methods will facilitate detection of new inhibitors and investigation of risk factors.
Epitope Identification and Generation of an Antigen-Specific T-Cell Clone in a Mild Hemophilia A Inhibitor Subject with Missense Genotype R593C.
Authors: James Eddie A. , van Haren Simon D. , Ettinger Ruth A. , Thompson Arthur R. , Kwok William W. , Voorberg Jan , Pratt Kathleen P. 
Published: October 2019

Abstract
The development of inhibitory anti-FVIII antibodies is a major clinical problem in hemophilia A. While less common in mild/moderately severe patients, the relative risk of inhibitor formation is elevated in patients with missense mutations in the FVIII A2 domain, especially those with an R593C genotype. In this study, T-cell responses to 14 FVIII A2 domain peptides with predicted DRB1*1101 MHC binding motifs were investigated using MHC class II tetramer reagents. CD4+ T cells were isolated from a hemophilic subject (DRB1*1101, 1302) with FVIII missense genotype R593C and stimulated with pooled peptides. This subject had developed a longstanding low titer inhibitor after receiving multiple FVIII infusions. Staining with fluorescent, peptide-loaded tetramers revealed that the hemophilic subject, but not an HLA-matched healthy control, had a DRB1*1101-restricted response to peptide A2589–608, which contained the wild-type R593 sequence. MHC class II tetramers with bound A2589–608 were used to sort antigen-specific T cells and then generate a T-cell clone recognizing this high avidity DRB1*1101-restricted epitope. FVIII residues 594–602 (FLPNPAGVQ) comprise a predicted high-affinity binding motif. Peptide binding assays confirmed that A2589–608 bound to recombinant, monomeric DRB1*1101 protein with high affinity. A peptide with the corresponding hemophilic sequence (A2589–608, 593C) bound with affinity that was threefold lower, but in a range that should be sufficient for presentation to T cells. The other 12 A2 peptides had a wide spectrum of binding affinities for the DRB1*1101 protein. Interestingly, prediction algorithms suggest that the peptides containing residue 593 occupy the MHC Class II peptide-binding groove in a register that places this residue just outside the p1 anchor position. Thus we anticipate that residue 593 modulates T-cell recognition of peptide A2589–608. This provides a potential explanation for inhibitor development in mild hemophilia A patients with a FVIII R593C genotype who are HLA-DRB1*1101. Immunogenicity of this FVIII region with respect to other HLA types is under investigation.