bluuebunny/crossref_metadata_2025_split huggingface dataset:
Defective Engraftment of HSPC from C3aR−/ − Mice Reveals an Underappreciated Role of C3a-C3aR Axis in Stem Cell Homing to Bone Marrow.
Authors: Reca Ryan , Kucia Magda , Baran Jarek , Ratajczak Janina , Ratajczak Mariusz Z. 
Published: October 2019

Abstract
Recently we demonstrated that complement (C) is activated in bone marrow (BM) during conditioning for transplantation and hematopoietic stem/progenitor cells (HSPC) express on their surface the C3a receptor (C3aR) and in the presence of the third C component - C3 cleavage fragments (C3a and desArgC3a) respond more robustly to a chemotactic gradient of stromal-derived factor (SDF)-1 (Reca et al., Blood2003, 101, 3784). The molecular explanation for this phenomenon is a C3a mediated increase in the incorporation of CXCR4 into membrane lipid rafts what enables CXCR4 to interact better with small GTPases from the Rho/Rac family (Wysoczynski et al. Blood2005, 105, 40–48). To elucidate this phenomenon better and to learn more on the role of the C3a-C3aR axis in homing/engraftment of HSPC we studied i) engraftment of murine HSPC derived from C3aR-deficient mice into wild type littermates and ii) human HSPC on which C3aR was blocked by C3aR antagonist SB290157 into NOD/SCID mice. We noticed that wt mice transplanted with C3aR−/ − HSPC engrafted significantly worse compared to normal littermates. Accordingly, transplantation of the same numbers of Sca-1+ cells from C3aR−/ − mice into wt littermates as compared to transplantation of wt cells resulted in i) delay by ~5–7 days in recovery of platelets and leukocytes, ii) decrease in day 12 CFU-S, and iii) decrease in the number of CFU-GM progenitors detectable in the BM cavities at day 16 after transplantation. Similarly in parallel experiments, human CD34+ cells exposed to nontoxic doses of C3aR antagonist SB29007 engrafted worse in NOD/SCID mice (p<0.0001). Next, we studied the different steps of homing of HSPC and noticed that sensitization of cells to an SDF-1 chemotactic gradient was compensated in C3aR−/ − mice probably by the activation of another putative receptor for C3a, however, the C3aR was indispensable for optimal adhesion of HSPC to endothelium and SDF-1-dependent MMP-9 secretion. In conclusion, activation of the C cascade in BM during conditioning for transplantation exposes a natural neoantigen which is recognized by immunoglobulins activating C by the classical pathway. As a consequence, C3 cleavage product, C3a, activates the C3aR on transplanted HSPC increasing the SDF-1 mediated homing of these cells.
Sensitive and Quantitative Measurement of Gene Expression Directly from Peripheral Whole Blood, without RNA Isolation and Target Amplification.
Authors: Zheng Zhi , Luo Yuling , McMaster Gary 
Published: October 2019

Abstract
Peripheral blood gene expression analysis is increasingly used for diagnosis and prognosis of hematological diseases, as well as for surrogate biomarker discovery in a wide range of non-hematological disorders. However, a critical issue concerning the potential clinical application of these research findings relates to the validity and reproducibility of the blood mRNA quantitation results. Current gene expression technologies often depend on multiple steps: 1) blood isolation; 2) RNA purification and 3) subsequent enzymatic reactions, which affect the accuracy and consistency of results. We describe an assay to measure single- and multi-plexed gene expression directly from whole blood without RNA purification and target amplification. The hybridization-based assay uses branched DNA signal amplification technology with a whole blood lysis protocol that preserves the RNA integrity. The assay is sensitive enough to quantitatively measure genes expressed at low levels in a minority of cells from less than 30ul of whole blood. The coefficients of variations are less than 10% and the dynamic range is 3–4 logs for both singleplex and multiplex formats. The assay signals are several times higher than purified RNA from an equivalent amount of blood. Blood proteins, genomic DNA and reticulocyte mRNAs do not interfere with the assay. The assays are compatible with common anticoagulants and Paxgene treated samples. However, we found the Paxgene induced expression of antiapoptotic genes during processing of the whole blood. We used the multiplex assay to evaluate the impact of common blood processing on the expression of a panel of 30 cytokine and apoptosis genes known to be sensitive to ex vivo purtabation. Minimal impact was found with RBC lysis, followed by whole blood RNA extraction, PBMC isolation and Paxgene stabilization. The lowest correlation of expression was found between RNA extracted from whole blood and RNA extracted from same blood treated with Paxgene (R square=0.77). We believe this assay will contribute to fundamental and therapeutic applications where quantitative gene expression and/or throughput are required.
Preliminary Phase II Results of SGN-30 (Anti-CD30 Monoclonal Antibody) in Patients with Cutaneous Anaplastic Large Cell Lymphoma (ALCL).
Authors: Duvic Madeleine , Kunishige Joy , Kim Youn H. , Foreno-Torres Andres , Pinter-Brown Lauren C. , Rarick Mark U. , Zic John , Worobec Sophie M. , Foss Francine , Flessland Karen , Barton Jeremy 
Published: October 2019

Abstract
Background: SGN-30 is a chimeric monoclonal antibody (mAb) directed against the CD30 antigen which is expressed on Hodgkins and non Hodgkins Lymphoma. SGN-30 has been well tolerated in phase I and phase II studies and objective responses have been seen in systemic ALCL.
Methods: Adult CD30 positive cutaneous ALCL patients who have progressed after treatment with radiotherapy and/or systemic therapy, ECOG performance status 0–2, are treated in a multicenter, single arm phase II study. SGN-30 at 4 mg/kg is administered by IV infusion monthly for a maximum of 6 consecutive doses. Evaluation of response (Physician’s Global Assessment) is performed 3–4 weeks after each infusion. In the absence of an objective response after two doses, patients may receive SGN-30 at 12 mg/kg for the remaining infusions.
Results: Patient Characteristics: 5 patients (4 male, 1 female) with median age 63 years (range 42–79) have been enrolled. Median number of prior therapies is 3 (range 1–4). All patients had at least 1 prior systemic therapy, and 3 of 5 patients (60%) had received at least two of the following therapies: Targretin, CHOP, or radiotherapy. 4 patients received 6 doses of SGN-30; of these 1 patient increased to 12 mg/kg at dose 3, 1 at dose 5, and 2 at dose 2. One patient received a single 4 mg/kg dose. Efficacy: 4 of 5 patients (80%) achieved a response; 1 CR and 3 PR (2 designated as “Marked Improvement” and 1 as “Almost Clear”). The patient with a CR had a history of Mycosis Fungoides and developed CD30+ ALCL. He had complete resolution of trunk lesions after 5 doses of SGN-30, received one additional dose of SGN-30, and maintained a durable CR of > 40 days. Of the 3 patients with PR, 1 attained a response after one dose, and 2 after 3 doses. One patient with concurrent LyP received a single dose of SGN-30 9 days after discontinuing methotrexate, experienced a worsening of his disease, and came off the study with PD. Tolerability: The most common adverse events in the 5 patients included pedal edema, arthralgia, and insomnia, each of which occurred in 2 patients. Of the 6 grade 3/4 adverse events, 4 were considered to be unrelated to SGN-30, and the relationship of 2 were unknown (pruritis and increased number of lesions). There was 1 SAE, an 80 year old male hospitalized with pneumonia after 6 doses of SGN-30 died on study of myocardial infarction secondary to aortic stenosis, considered unrelated to study drug.
Conclusions: Preliminary data indicate the drug is well tolerated and shows antitumor activity. Recruitment into the study continues.
Type of BCR-ABL Transcript Predicts Response to Imatinib Mesylate.
Authors: Sharma Pratibha , Kumar Lalit , Kochupillai Vinod , Mohanty Sujata , Sharma Atul , Seth Tulika 
Published: October 2019

Abstract
Imatinib Mesylate, a molecular target therapy is the first line treatment for all CML patients. We prospectively studied 50 CML patients (37 pre-treated with interferon Alfa) for response to Imatinib. Patients median age was 32 years, range, 11 to 62 years with male to female ratio being 2.5: 1. All patients were in chronic phase and received 400 mg imatinib daily for a mean time period of 18 months (range 3 to 38 months). Patients were monitored closely for clinical, hematological, cytogenetic & molecular response. Bone marrow examination was done at 3, 6, 9 then 4–6 months interval for cytogenetic response (CGR) and also for presence of BCR-ABL transcript types (b2a2, b3a2, e1a2 and e19a2) by multiplex and nested RT-PCR technique. All patients achieved complete hematological remission with in 3 months of starting Imatinib. 20/50 patients achieved Major CGR (complete -8, Partial CGR-12). Of these, 12/20(60%) patients had b2a2 transcript, 3/20 (15%) b3a2 transcript and 5/20 (25%) had both the transcripts. 24/50 had minor CGR (b2a2-10/24 (41.6%), b3a2-14/24 (58.3%). Remaining 6 patients had no Cytogenetic response (4/6-b3a2, 2/6-b2a2). Patients with BCR-ABL transcript b2a2 had a significant probability of achieving Major cytogenetic response 12/50(24%) compared with b3a2 transcripts, 3/50 (6%), p<0.001. However, there was no significant correlation between transcript type and spleen size, Hb, WBC, platelets counts. These initial observations are very provocative and suggest that site of breakpoint in major breakpoint cluster region might be predictive of cytogenetic responses to Imatinib Mesylate. We are continuing to accrue more patients in this study to confirm these results.
ATP Binding Domain Mutants of BCR-ABL in Patients with Chronic Myeloid Leukemia Treated with Imatinib.
Authors: Liu Xiaoli , Zhang Song , Du Qingfeng , Fan Wei , Wang Yu , Yao Feng 
Published: October 2019

Abstract
Objective:Imatinib (also called STI571 or Gleevec), a competitive inhibitor at the ATP binding site of BCR-ABL, has been shown remarkable clinical activity in patients with chronic myeloid leukemia (CML). However, a significant proportion of Imatinib-treated CML patients with advanced stage disease develop resistance. In this study, we try to detect ATP binding domain mutants of BCR-ABL in Imatinib-treated CML patients and explore the possibility that ATP binding domain mutants might confer resistance to imatinib.
Patients and Methods:We analyzed bone marrow samples from 16 Imatinib-treated CML patients, including 6 chronic phase, 2 accelerated phase (AP) and 8 blast crisis (BC) patients. 8 patients were sensitive to imatinib and 8 patients resistant. A polymerase chain reaction strategy was used to amplify and sequence the ATP binding domain of BCR-ABL.
Results:Point mutations were found in the ATP binding domain of BCR-ABL in 3 of 16 patients. In 2 of 8 imatinib-resistance patients, a single nucleotide change was detected and resulted in a threonine to isoleucine substitution at position 315(Thr315Ile) of ABL kinase. This point mutation has proved to interfere with drug binding and cause resistance to imatinib. In addition, we found a mutation (Lys357Arg) remain sensitive to imatinib.
Discussion:Chronic myeloid leukemia (CML) is a hematopoietic disorder characterized by the malignant expansion of bone marrow stem cells. Its malignant clonal marker is Philadelphia chromosome (Ph) which harbors the BCR-ABL fusion gene. The latter encodes a chimeric BCR-ABL protein, identified as having a central role in the pathogenesis of CML. As a competitor for ATP binding of BCR-ABL, Imatinib selectively induces apoptosis and blocks proliferation in BCR-ABL positive cells and has been shown to have high activity in CML. Clinical studies demonstrate that Many Imatinib-treated CML patients with advanced stage disease respond initially but then relapse. Till now, the resistant mechanisms of imatinib have not all been known. In this study, we find that ATP binding domain mutants of BCR-ABL is one of the potential mechanisms of resistance to imatinib, but not all mutants associate with imatinib resistance and some mutants remain sensitive to imatinib.
Hematopoietic Precursor Cell Cultures (CFU-C) in Diagnosis and Prognosis of Myelodysplastic Syndromes (MDS) and Chronic Myelomonocytic Leukemia (CMML).
Authors: Bernimoulin Michael Pierre , Jotterand Martine , Tichelli Andre , Meyer-Monard Sandrine , Heim Dominik , Gratwohl Alois , Nissen Catherine 
Published: October 2019

Abstract
Background
Diagnosis and prognosis of patients with established MDS/CMML is currently based on WHO criteria and the IPSS prognostic Index. High variation of survival in subgroups warrants the search for additional criteria. A comparison of CFU-C cultures with WHO criteria and IPSS score has not yet been done in a large patient group.
Patients and methods
We analyzed in a single center retrospective cohort study 93 untreated consecutive patients (55 male/ 38 female; median age: 66 years; range: 13 – 88 years) admitted between July 1992 and June 2002 and diagnosed as MDS/CMML (RA/RARS(4), MDS-U (2), RCMD (26), RAEB I/II (44) and CMML I/II (17). All patients had an unequivocal diagnosis of MDS or CMML according to WHO criteria. Simultaneous examinations of blood, bone marrow (cytology and biopsy), BM-cytogenetics and BM-and PB cultures for CFU-GM and BFU-E were done. Culture results were scored blindly and classified either as “Low risk CFU-C“ including normal growth (N=2), no colony growth (N=6) or reduced growth of normal colonies (N=19) or as „High risk CFU-C’s“ including excess normal growth in PB termed “MPS pattern” (N=5), discrete leukemic cluster growth (N=22), abundant leukemic cluster growth (N=14) or the „CMML pattern“ defined as giant“ pseudonormal“ CFU-GM and strongly reduced BFU-E (N=23). Culture score was compared with the WHO diagnosis and with the IPSS score in all 93 patients and survival was assesed in 82 patients treated with conventional therapy (12 allografted patients were excluded) after minimal observation time of 3 years in July 2005.
Results
Comparison of WHO diagnosis with culture score Low risk CFU-C score High risk CFU-C score RA/RARS/RCMD/MDS-U 15 16 RAEBI/II 12 30 CMML-I/II 0 17
The typical CMML pattern was observed in 13 of 17 patients with CMML (Specificity 94%).
Comparison of IPSS Score with culture score low risk CFU-C score low risk CFU-C score High risk CFU-C score High risk CFU-C score IPSS Score alive/dead mean survival(d) alive/dead mean survival(d) Low and Int-1 4/4 2022+/−543 8/39 965+/−143 Int-2 and High 1/1 1024+/−701 1/26 565+/−92
Mean survival time for patients with a low/intermediate-1 IPSS score was less than half if they had a high risk CFU-score (p<0.01, ANOVA t-test). For patients with an intermediate-2/high IPSS score the prognostic information gained with the cultures was similar, but low patient number did not permit statistical analysis.
Conclusions
Hematopoietic precursor cultures yield additional information in patients with MDS / CMML. For diagnosis, CMML has a typical growth pattern, that may help distinction of clonal from reactive monocytosis. For prognosis, the culture pattern adds information to the IPPS score, leukemic growth being strongly predictive of short survival.
Telomerase and Telomeres in Polycythemia Vera and Essential Thrombocythemia.
Authors: Gallamini Andrea , Ferraris Anna Maria , Mattei Daniele , Pujic Natalija , Mangerini Rosa , Rapezzi Davide , Tavera Sivia , Gaetani Gian Franco 
Published: October 2019

Abstract
Polycythemia Vera (PV) and Essential Thrombocythemia (ET) are chronic myeloproliferative disorders (CMPD) with a relatively favorable prognosis and a long natural history. They originate from a multipotent progenitor cell with dominance of the transformed clone over normal hematopoiesis. However, the heterogeneity of these diseases with respect to clonal development from a common progenitor has been established, and no specific molecular or cytogenetic markers are so far considered useful prognostic indicators. In the current study we correlate telomerase activity (TA) and telomeres [terminal restriction fragment (TRF)] lengths in granulocytes (PMN) and mononuclear cells (MNC), with the results of the clonality status, as determined by investigation of X chromosome inactivation patterns (XCIPs), from 22 informative female patients with ET and PV. All patients were in chronic phase without immature myeloid cells in the peripheral blood. After testing for heterozygosity at the HUMARA locus PMN expressed a monoclonal XCIP in 9 out of 13 subjects with ET (69%), and in 6 out of 9 with PV (67%). MNC were always polyclonal. The amount of TA for each reaction was expressed in Total Product Generated (TPG) units. ΔTRF length was calculated by subtracting the telomer length of PMNs from that of MNCs and expressed as kilobases. TA and ΔTRF results were blindly compared with those from XCIP analysis. Monoclonal PMN displayed a TA ranging between 47.6 and 212.5 TPG units (mean 118.2 ± 45.5 SD), whereas polyclonal PMN had a TA between 1.0 and 33.3 TPG units (mean 14.4 ± 11.9 SD) (p<0.001). MNC had a TA similar to that of polyclonal PMN. Mean ΔTRF of clonal PMNs was 4.6 kilobases, while that of polyclonal PMNs was 0.44 kilobases (p<0.001). PMNs from normal control subjects had a mean ΔTRF of 0.1 kilobases, similar to that of polyclonal PMNs (p= n.s.). The results of our study clearly show the existence of two distinct subgroups of CMPD, with polyclonal or monoclonal granulopoiesis, each of them correlating with a peculiar pattern of TA and telomere lengths. Although none of the patients analyzed showed any difference in clinical characteristics with respect to the parameters considered, we believe that TA and ΔTRF analysis and their relationship with clonality status may represent an useful tool for further investigation, to predict survival in larger cohorts of patients with PV and ET, over an extended period of follow up.
A Comparison of Zap-70 by Immunohistochemistry and Flow Cytometry Using Two Different Flow Analysis Methods.
Authors: Hertzberg Lawrence , Baggianni Karina , Tamayo Rosalba , Browne Patrick , Oldaker Teri 
Published: October 2019

Abstract
Zap-70 expression by flow cytometry has been shown to be a surrogate marker for un-mutated/germline IgH status in chronic lymphocytic leukemia (CLL).1 Reproducibility and confidence in the accuracy of results can make this assay problematic. We compared our current single-parameter/one-color, immunohistochemistry (IHC) Zap-70 assay with our flow cytometry Zap-70 assay. Twenty-one (21) patient samples submitted to our lab for Zap-70 testing were assayed by IHC and flow cytometry (FC). In addition, two FC analysis methods were used. One analysis method uses isotype controls to establish negative regions and a 20% cut-off for positivity1, with BD CellQuestTM software (CQ). The other analysis method uses internal negative and positive mean channel fluorescence (MCF) differences using BD Paint-A-GateTM software (PAG). The IHC assay resulted in nine positive, six negative and six equivocal results for Zap-70. The equivocal results were due to very low proportions of CLL cells (three cases) or other problems identified on review of stained slides (three cases). The FC assay resulted in six positive, thirteen negative and two equivocal results for both analysis methods. When comparing IHC Zap-70 and FC Zap-70, there was 29% (6/21) discordance using the CQ analysis and 24% discordance (5/21) using the PAG analysis (equivocal cases included in the denominator only). Comparison of the CQ and PAG analysis demonstrated 16% (3/19) discordance (equivocal cases excluded from the denominator). The FC Zap-70 PAG analysis method correlated somewhat better with the IHC Zap-70 results than did the FC Zap-70 CQ analysis method. This may be due to the variability introduced by user-defined/isotype cursor settings. We conclude that Zap-70 expression can be tested for using either IHC or FC, and that IHC testing may have higher sensitivity. Correlation between the results utilizing these different methods is not ideal, and improvement in both assays, likely including flow analysis methods, is needed. We plan additional, comparative evaluation of more cases.
CellQuest™ and Paint-A-Gate™ are trademarks of Becton, Dickinson and Company.
The Role of Proteasome Inhibitors and the Trail Apoptotic Pathway in the Treatment of Chronic Lymphocytic Leukemia.
Authors: McCrea Kristin E. , Kabore Albert , Johnston James B. , Gibson Spencer B. 
Published: October 2019

Abstract
TRAIL (tumour necrosis factor-related apoptosis-inducing ligand) triggers the TRAIL apoptotic pathway selectively in tumour cells by binding to two death receptors, DR4 and DR5, that are present on the surface of many cancer cells. It is effective at inducing apoptosis in a variety of haematological malignancies, such as multiple myeloma. However, chronic lymphocytic leukemia (CLL) cells are relatively resistant to TRAIL-induced apoptosis. We have previously shown that the chemotherapeutic drugs, fludarabine and chlorambucil, increase the cell surface expressions of DR4 and DR5, and give synergistic apoptotic responses when combined with TRAIL. Proteasome inhibitors, that are used in the treatment of multiple myeloma, also upregulate TRAIL and its death receptors in CLL cells but not in normal B cells. Herein we have determined that proteasome inhibitors are effective at inducing apoptosis in CLL cells, and that the activation of the TRAIL apoptotic pathway contributes significantly to proteasome inhibitor cytotoxicity. Combining proteasome inhibitors with TRAIL enhanced apoptosis in CLL cells by approximately 15% over treatment with the proteasome inhibitor alone. Another novel approach to trigger the TRAIL apoptotic pathway is to use activating monoclonal antibodies directed against DR4 and DR5. Similar to TRAIL, we demonstrated that when used alone, the monoclonal antibodies were minimally cytotoxic against CLL cells. Proteasome inhibitors in combination with activating monoclonal antibodies against DR4 or DR5 increased the amount of apoptosis in CLL cells. Cell death was enhanced by 23% and 17% when proteasome inhibitors were combined with monoclonal antibodies against DR4 and DR5, respectively. While proteasome inhibitors may have a potential role in CLL treatment as single therapy, they are also cytotoxic to normal B cells. However, when activating monoclonal antibodies against TRAIL death receptors are given in combination with proteasome inhibitors, that upregulate DR4 and DR5 expression, the anti-tumor specificity and cytotoxicity of these agents may be increased in CLL.