bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prognostic Value of Early 18fluorodeoxyglucose PET Scan in Patients with Diffuse Large B Cell Lymphoma Treated with Rituximab Plus CHOP or High-Dose CHOP.
Authors: Dupuis Jehan , Itti Emmanuel , Fruchart Christophe , Bardet Stéphane , Rahmouni Alain , Vilque Jean-Pierre , Lin Chieh , Belhadj Karim , Gnaoui Taoufik El , Kuhnowski Frédérique , Meignan Michel , Haioun Corinne 
Published: April 2020

Abstract
The results of 18Fluorodeoxyglucose – Positron Emission Tomography scan (PET) performed early during treatment (after 1 – 3 courses of chemotherapy) have been shown to predict outcome in aggressive lymphoma (Jerusalem, Haematologica 2000; Spaepen, Ann Oncol 2002; Haioun, Blood 2005). However, studies on early PET have been performed mostly in patients (pts.) treated without Rituximab. Our goal was to explore if the powerful predictive value of early PET was also observed in pts. treated with Rituximab-chemotherapy combinations. Seventy-three pts. with diffuse large B-cell lymphoma from 2 institutions were treated with R-CHOP (n=47) or R-high-dose CHOP (R-ACVBP, n=26) and were evaluated by PET after the two first cycles. PETs were interpreted on the basis of visual assessment. PET results were not taken into account to guide the treatment strategy after the 2 first cycles, which was solely based upon the results of conventional response assessment. Among responders, 16 young poor-risk pts. received planned consolidative high-dose therapy with autotransplant, while the others received immunochemotherapy based consolidation. Median age was 63 years (range 26–79) and 68% pts. were male. Baseline characteristics were as follows: Advanced Stage (Ann Arbor III–IV) 75%; Elevated LDH 62%; Poor performance status (ECOG 2–4) 32%. The International Prognostic Index was as follows: Low risk 27%, Low-intermediate 21%, Intermediate-high 21%, High risk 31%. PET was performed after a median of 15 days following the second cycle. Forty-nine pts. had a negative PET, while 24 were positive. With a median follow-up of 49 months, 11/49 (22%) PET negative pts. showed an event (progression or death) after a median delay of 13 months, while this was the case in 14/24 (58%) PET-positive patients after a median of 8 months. The estimated 5 year event free survival rates were 75% (95% CI: 63%–88%) for PET negative pts. and 41% (95% CI: 22%–61%) (log-rank test, p=0.001). Regarding overall survival, the estimated 5 year rates were 85% (95% CI: 78%–95%) and 53% (95% CI: 34%–74%) (log-rank test, p=0.005) in PET negative and PET positive pts., respectively. The same differences were found at 4 years when analysing separately pts. treated with R-CHOP and R-ACVBP (except for a non significant difference in OS in R-ACVBP pts.). In conclusion, we confirm the strong prognostic value of PET after 2 cycles in pts. treated with Rituximab combined to chemotherapy. Prospective studies (including a GELA study) are ongoing to evaluate the potential interest of tailoring treatment strategy to the results of early PET.
Mapping Interactions Between B2GPI and the Lipoprotein Receptors by Solution NMR Spectroscopy.
Authors: Beglova Natalia , Lee Chang-Jin 
Published: April 2020

Abstract
Antiphospholipid syndrome (APS) is an autoimmune disease characterized by venous or arterial thrombosis and recurrent pregnancy loss. Beta2 glycoprotein I (B2GPI) is the major target for these antibodies. Current evidence suggests that B2GPI acquires pathological properties leading to APS after association with antibodies. It was demonstrated that APS related antibodies induce activation of endothelial cells, monocytes and platelets, although the identity of a cell-surface receptors that transmit the antibody-induced signaling inside cells remain unclear. Biochemical studies have shown that the receptors of the low-density lipoprotein receptor (LDLR) family are all capable to bind B2GPI/antibody complexes. These receptors are abundantly expressed by many cell types. The ligand-binding activity of the lipoprotein receptors is confined primarily to structurally homologous LA modules, which are arranged in sequence to form a ligand-binding domain. We investigated at the atomic level the features of the ligand-binding LA modules required for interaction with B2GPI. We sought to determine whether the same residues that form the ligand-binding pocket on the LA modules in the crystal structure of a RAP/LA3-4 complex also recognize B2GPI. We demonstrated that the same residues that form the ligand-binding pocket on LA4 in the crystal structure of the RAP/LA3-4 complex comprise the contact site for B2GPI. The domain 5 from B2GPI (B2GPI-D5) was expressed and labeled for NMR studies. The residues of B2GPI-D5 that are perturbed when B2GPI-D5 was titrated with LA4 are located at the C-terminus of domain 5. Using NMR relaxation measurements, we determined that B2GPI-D5 binds LA4 with 1:1 stoichiometry. The estimated Kd of the B2GPI-D5/LA4 complex is about 30 uM. NMR experimental data was used to build a docking model of the complex.
Protease Activated Receptor-4 (PAR4) Uses Anionic Residues to Interact with α-Thrombin in the Absence or Presence of PAR1.
Authors: Nieman Marvin T 
Published: April 2020

Abstract
Thrombin activates protease activated receptor 1 (PAR1) faster than protease activated receptor 4 (PAR4) due to a hirudin-like sequence in the exodomain of PAR1 that binds thrombin’s exosite I. However, recombinant exodomain studies indicate that PAR4 does have extended contacts with α-thrombin that influence PAR4’s kinetics of cleavage. In the current report, the role of an anionic cluster (Asp57, Asp59, Glu62, Asp65) in the exodomain of PAR4 is examined for its influence on cleavage and activation of PAR4 on cells in the absence or presence of PAR1. α-Thrombin induces wild type PAR4 (PAR4-wt) calcium flux with an EC50 of 110 nM whereas mutation of the four anionic residues (PAR4-AAAA) increases the EC50 to 641 nM. In contrast, PAR4-wt and PAR4-AAAA are activated by γ-thrombin with a similar EC50 (588 nM and 449 nM, respectively, p = 0.48), indicating a role for α-thrombin’s exosite I in PAR4 activation. Coexpression of PAR1, lowered the EC50 of cleavage 10 fold for both PAR4-wt from 321 to 26 nM and PAR4-AAAA from 2.2 μM to 360 nM, respectively. Individual point mutations at Asp57, Asp59, Glu62 or Asp65 show that PAR4-D57A is activated by α-thrombin with the same EC50 as PAR4-wt (140 nM) whereas PAR4-D59A is the same as PAR4-AAAA (699 nM). Glu62 and Asp65 contribute to α-thrombin recognition, but to a lesser extent. The current report shows that PAR4 uses its anionic cluster to interact with α-thrombin and that this interaction is important even in the presence of PAR1.
Comparative Effects of the Direct Factor Xa Inhibitor, Apixaban, and the Direct Inhibitors of Factor VIIa, XIa and Thrombin on Tissue Factor- Induced Human Platelet Aggregation.
Authors: Jiang Xiaosui , Wong Pancras C. 
Published: April 2020

Abstract
Background: Apixaban is an oral, direct and highly selective factor Xa (FXa) inhibitor in late-stage clinical development for the prevention and treatment of venous thromboembolism, stroke prevention in patients with atrial fibrillation, and secondary prevention in patients with acute coronary syndrome. By inhibiting FXa, apixaban has the potential to reduce thrombin generation and indirectly decrease thrombin-mediated platelet aggregation. This study evaluated the in vitro effect of apixaban on human platelet aggregation induced by thrombin derived via the extrinsic pathway. Direct inhibitors of FXa (rivaroxaban), FVIIa (BMS-593214), thrombin (dabigatran, argatroban) and FXIa (BMS-262084) were included for comparison.
Methods: Citrated human blood, mixed with corn trypsin inhibitor (50 μg/ml) to block the contact factor pathway, was centrifuged to obtain platelets-rich plasma (PRP). The latter were incubated for 2 min at 37°C with 3 mM H-Gly-Pro-Arg-Pro-OH-AcOH (to prevent fibrin polymerization) and with vehicle or increasing concentrations of inhibitors. Human tissue factor (TF) (Innovin®; dilution 1:900 to 1:1000) plus 7.5 mM CaCl2 was added to PRP to trigger platelet aggregation. Platelet aggregation response, measured turbidometrically with a platelet aggregometer, was expressed as AUC occurring 5 min after TF stimulation.
Results: TF produced 85±3% aggregation of human platelets in the vehicle-treated group (n=10). Apixaban and other factor inhibitors, except the FXIa inhibitor, inhibited TF-induced platelet aggregation in a concentration-dependent manner with IC50 (nM) values as follows: 3.6±1.3 (apixaban), 7.5±1.5 (rivaroxaban), 13±1 (BMS-593214), 46±1 (dabigatran) and 79±1 (argatroban). BMS-262084 (IC50 = 2.8 nM vs. human FXIa) had no effect on TF-induced platelet aggregation at 10 μM. These inhibitors at 10 μM had no effect on platelet aggregation induced by ADP and collagen, as expected from their mechanism of action.
Conclusions: This study demonstrates that inhibition of thrombin generation by blocking upstream proteases (FVIIa and FXa) in the blood coagulation cascade is as effective as direct thrombin inhibition in preventing TF-induced platelet aggregation. Under these experimental conditions, FXIa did not play a role in platelet aggregation. Further, the potent indirect antiplatelet effect of FXa inhibitors together with their anticoagulant effects suggest that apixaban may have therapeutic potential in patients with arterial thrombosis. Accordingly, apixaban is in development for secondary prevention in patients with acute coronary syndrome.
Detection of Minimal Residual Disease in Chronic Lymphocytic Leukemia with Monoclonal Antibodies Specific for CD5, CD10, CD19, and ROR1.
Authors: Broome H. Elizabeth , Rassenti Laura , Bui Jack D. , Meyer Lilly , Kipps Thomas J. 
Published: April 2020

Abstract
ROR1 is an oncofetal protein expressed in chronic lymphocytic leukemia (CLL). We generated a monoclonal antibody (mAb) specific for ROR1 (4A5) and used this to stain blood or marrow cells from patients with CLL and other adults. We found that 4A5 reacted with the CLL cells of all patients examined and also with some cells that had an immunophenotype of B lymphocyte precursors (BLP) (CD10+, CD19+, CD20-variable, dim CD45+, surface immunoglobulin-negative). 4A5 showed no reactivity with other blood or marrow cells. We speculated that this mAb could be used to detect CLL cells in blood or marrow of patients after therapy, providing a means with which to assess minimal residual disease (MRD). For this purpose, we evaluated a four-color combination of mAbs using CD10-FITC, CD19-PE, CD5-PerCP-Cy5.5, and 4A5-Alexa-647. MRD detection limits were established through reconstitution studies in which we made serial dilutions of CLL cells into normal blood or marrow samples. MRD was measured by whole blood lysis and acquisition of 100,000 events using a FACSCalibur and a gating strategy designed to exclude CD5-negative, CD10-positive, and 4A5-negative B lymphocytes (CD19+). CLL cells stained with 4A5 had a mean fluorescence intensity (MFI) of 31 (N=100) and a MFI ratio (MFIR) relative to that of isotype control stained CLL cells of more than 10:1. On average, 94% of the CLL cells in each sample had higher fluorescence intensity when stained with 4A5-Alexa-647 than when stained with an isotype-control mAb (range 88–98%). Marrow samples (N=9) from adults with diagnoses other than CLL and that had 1–10% BLP had CD5− negative, CD10+, CD19+ cells that reacted with the 4A5-Alexa 647. These cells in such samples had a MFI of 19 and a MFIR relative to that of isotype-control-stained BLP of 6.3. On average, 48% of such cells in each sample had higher fluorescence intensity when stained with 4A5-Alexa-647 than when stained with an isotype-control mAb (range 18–79%). Four color flow cytometric analysis with CD10, CD19, CD5, and 4A5 detected CLL cells present at less than 0.1% in reconstituted blood or marrow samples, including marrow with 3–5% BLP. Background was between 0.01% and 0.1% with whole blood (marrow) lysis when we acquired a total of 100,000 flow-cytometric events. These data indicate that anti-ROR1 mAbs can be used with mAbs specific for CD10, CD19, and CD5 for sensitive detection of MRD in CLL.
Prognostic Stratification of Patients with Chronic Lymphocytic Leukemia Based on Quantitative Flow Cytometry Evaluation of ZAP-70 Versus Homology of IgVH Plot.
Authors: Klabusay Marti n , Kuhrova Viera , Hrabcakova Viera , Coupek Petr , Mayer Jiri 
Published: April 2020

Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is the most common type of leukemia in the adult population. It has a heterogeneous behavior and variable prognosis. While some patients experience indolent disease requiring no therapy for many years, others demonstrate a more aggressive type unresponsive to therapy. An accurate prognostic stratification is essential for optimizing the therapeutic strategy. Many prognostic factors are presently known, but their relationships and significance are often not clearly understood. Several markers have been identified, including IgVH mutation status and ZAP-70. Mutation of IgVH (less than 98% homology with the germline sequence) is correlated with better prognosis. Expression of ZAP-70, which is a cytoplasmic ζ-associated tyrosine-kinase essential for T-cell receptor signal transduction, is associated with more rapid disease progression and shorter survival. However, its detection by flow cytometry has many technical difficulties, resulting in high interlaboratory variability. The expression of intracellular ZAP-70 in 217 patients with diagnosis of B-CLL was determined using a new approach: quantitative flow cytometry on CD5+19+ tumor cells. Other laboratory and clinical parameters were evaluated, including gender, age, type and number of therapies, CD38 expression, cytogenetics and mutation status of IgVH. The expressions of ZAP-70 and CD38 were measured in molecules of equivalent soluble fluorochrome (MESF units). The results were correlated with mutation status of IgVH. Patients were divided into two groups by cluster analysis in a plot of IgVH homology versus ZAP-70 MESF (left panel). Overall survival curves for these groups are shown (right panel). It could follow that patients above the diagonal (triangles, group B) in the IgHV versus MESF plot have significantly poorer prognoses (p=0.0001) than do patients below this diagonal (circles, group A). Expression of CD38 above 15000 MESF showed worse prognosis for patients in the group B (p=0.035). Only three of 61 patients with chromosomal aberrations associated with poor prognosis (del11q and del17p) were found in the group A, but all of them had del13q present, as well. The authors introduced a new approach for evaluating ZAP-70 expression in B-CLL cells using quantitative flow cytometry that is easily standardized and not burdened with technical problems. Plotting IgVH homology (%) versus ZAP-70 quantitative expression (MESF) could clearly divide patients according to their prognoses.
Figure Figure
Nucleophosmin (NPM) Has Increased Expression and Shuttling Activity in Chronic Lymphocytic Leukaemia without Somatic Hypermutation of the Immunoglubulin Gene (UM-CLL) and Is Linked to Ribosomal Proteins: A Possible Role for Increased Protein biosynthesis in the Adverse Outcome of UM-CLL.
Authors: Rees-Unwin Karen S , Faragher Robin , Hutchinson Claire V , Unwin Richard D , Adams Julie , Pettitt Andrew , Banham Alison H , Pulford Karen , Burthem John 
Published: April 2020

Abstract
Chronic lymphocytic leukaemia (CLL) is a neoplastic proliferation of mature B-lymphocytes that has a marked variability of clinical outcome. Two distinct groups of CLL may be distinguished by the presence (M-CLL) or absence (UM-CLL) of somatic hypermutation of the immunoglobulin heavy-chain gene. UM-CLL has an adverse outcome, but the precise biological differences that underlie this more aggressive clinical-course are unclear. Using a proteomic approach, we have found that M-CLL and UM-CLL are consistently distinguished according to protein expression pattern. Of those proteins, the most important predictor of CLL class was the protein nucleophosmin (NPM). A significant increase in expression of NPM was found in UM-CLL compared with M-CLL (3.2 fold p&lt;0.05 Mann Whitney U test). This increased expression did not reflect differences in apoptosis or proliferation, or gene mutation. However, co-immunoprecipitation experiments identified the ribosomal proteins 60s (L19 and L24) and 40s (S9) to be associated with NPM in CLL. Ribosomal proteins are known to shuttle between the nucleoli, nucleus, and cytoplasm, of cells. Standard- and confocal-immunofluorescence microscopy identified NPM to be present, as expected, in the nucleoli of all CLL cells. Additionally however, NPM was consistently detected in the nucleoplasm of CLL, and in a proportion of cells from all CLL cases, NPM was also shown to be present in the cytoplasm (range =7–52% of cells expressed cytoplasmic NPM). Morphometric analysis suggested no net transfer from the nucleolus to the cytoplasm; but did suggest trafficking from the nucleoplasmic compartment to the cytoplasm consistent with shuttling between these two cellular regions. In UM-CLL the fluorescent intensity of the cytoplasmic NPM was found to be higher than that of the M-CLL (t-test p=0.009), and there was an approximate two fold increased proportion of cytoplasmic NPM-expression in the UM-CLL subgroup (p value&lt; 0.001). We propose therefore that NPM, in association with ribosomal proteins, displays high nucleocytoplasmic shuttling in CLL, and may reflect increased protein biosynthesis. In particular, the higher levels of NPM expression and shuttling in UM-CLL may play a role in the adverse outcome of this form of the disorder.
Predicting Time to Initial Treatment in Early Stage CLL - Integration of Classical and Novel Prognostic Factors.
Authors: Keating Michael J. , Lerner Susan , Wang Xuemei , Rassenti Laura Z. , O’Brien Susan , Kipps Thomas J. , Wierda William G. 
Published: April 2020

Abstract
CLL patients (pts) who are not eligible for initiation of treatment by the NCI Working Group criteria (1996) are traditionally monitored on a “watch and wait” approach. There is increasing interest in initiating therapy earlier in pts who are considered high risk. The most important advice pts require in the “watch and wait” category is whether they will need treatment and when. There are a number of traditional prognostic factors that potentially identify those “watch and wait” (W&W) pts most likely to need treatment. These have recently been amplified by the discovery of new prognostic factors such as the immunoglobulin (IgVH) mutation status, FISH cytogenetics, ZAP-70, and CD38 expression on CLL cells. Most reports have emphasized the new prognostic factors without consideration of traditional characteristics such as tumor burden, stage, etc. Since January 2004 FISH, IgVH mutation status, ZAP-70, and CD38 were characterized for most pts presenting to M. D. Anderson Cancer Center. We identified an early stage low tumor burden group of 826 pts (W&W) defined with as having lymph nodes &lt; 3 cm in the longest dimension, splenomegaly &lt; 5 cm, and Rai stage 0 – II. Twenty-four percent of these pts have received treatment in the first two years of follow-up. From the traditional prognostic factors, number of lymph node sites (cervical, axillary, and inguinal), absolute lymphocyte count (ALC), serum beta-2-microglobulin (B2M) levels were highly correlated with time-to-initial treatment (P&lt;.001). The predictive power of these characteristics was also confirmed in an earlier historic patient population. When these three characteristics are evaluated in multivariate analysis, IgVH mutation status, ZAP-70, and FISH cytogenetics all add independent prognostic power. When these three novel characteristics were evaluated in multivariate model with the traditional characteristics, the most important were the number of node sites, ALC, B2M, FISH, IgVH mutation status, and then ZAP-70.
Clinical only +Mutation Status +ZAP 70 +FISH + All 3 No. Lymph Node Sites 1 1 1 1 1 ALC 2 2 2 2 2 Beta-2-microglobulin 3 4 4 4 4 Mutation Status -- 3 -- -- 5 ZAP-70 -- -- 3 -- 6 FISH -- -- -- 3 3
Integration of novel prognostic factors into readily clinically available characteristics will enable us to develop a nomogram for predicting probability of each patient requiring therapy at one year, two years, and eventually five years. This nomogram will be presented when completed and then will be available for validation in independent patient populations studied by other groups.
CFAR, An Active Frontline Regimen for High-Risk Patients with CLL, Including Those with Del 17p.
Authors: Wierda William G , O’Brien Susan M , Faderl Stefan H , Ferrajoli Alessandra , Koller Charles , Estrov Zeev , Burger Jan A , Lerner Susan , Kantarjian Hagop M. , Keating Michael J 
Published: April 2020

Abstract
High-risk patients (pts) with CLL can be identified by prognostic factors. Traditional prognostic factors that identify high-risk pts include Rai stage, absolute lymphocyte count (ALC), rapid lymphocyte doubling time, and serum beta-2 microglobulin (β2M). Del 17p is associated with loss of the tumor suppressor gene TP53. Patients with del 17p also tend to have mutations in TP53, thereby providing a second mechanism for complete TP53 knockout. Pts with del 17p have poor response to purine analogue-based treatments. β2M greater than twice the upper limit of normal (2XULN) was an independent high-risk feature for pts treated with the frontline fludarabine (F), cyclophosphamide (C), rituximab (R) regimen (FCR). Pts younger than 70 years with β2M &gt;2XULN treated with FCR had a complete remission (CR) rate of 60% and median time to progression (TTP) of 70 months, compared to 84% and 86 months for pts &lt;70 yrs old with β2M &lt;2XULN. We evaluated the CFAR regimen (F 20mg/m2 d3–5; C 200mg/m2 d3–5; R 375–500mg/ m2 d2; and Alemtuzumab 30mg d1,3,5 of each 4-wk course) as frontline treatment in a high-risk group of pts with CLL. The trial completed planned enrollment of 60 pts; 48 are currently evaluable for response and follow-up. In this high-risk group, 28% of the 48 evaluable high-risk pts had del 17p prior to treatment by FISH analysis. Overall, the CR rate for the 48 pts was 69%, 54% CR for the 13 pts with del 17p. The overall response rates were 94% and 77% for the 48 pts and 13 pts with del 17p, respectively. CFAR was associated with more myelosuppression and fewer pts could receive all 6 intended courses compared with the historic high-risk group treated with FCR. However, a higher proportion of CFAR pts had no evidence of residual disease in the bone marrow by 2-color flow cytometry evaluation at response assessment. There is currently no difference in TTP or OS, comparing CFAR and FCR in this retrospective historical analysis with a short median follow-up time of 16 months for the CFAR group. There was no significant difference in incidence of infection during treatment with CFAR compared to FCR, with the exception of CMV reactivation. The CFAR regimen is active and tolerated in the highrisk group of pts with CLL, including those with del 17p. Follow-up continues for the pts treated on this trial in order to evaluate responses in all 60 enrolled pts and to evaluate time-to-event endpoints and compare with the historic FCR experience.