bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular Follow-Up of Patients Treated with HSV-TK/ΔLNGFR Engineered Donor Lymphocytes after Allogeneic Stem Cell Transplantation.
Authors: Bonini Chiara , Magnani Zulma , Recchia Alessandra , Urbinati Fabrizia , Muraro Sara , Tech Serenella Sartori , Ciceri Fabio , Bernardi Massimo , Pescarollo Alessandra , Gallo-Stampino Corrado , Bregni Marco , Bordignon Claudio , Mavilio Fulvio 
Published: October 2019

Abstract
Donor lymphocyte infusions (DLI) play a crucial role in promoting immune reconstitution and anti-tumor activity in patients treated with allogeneic hematopoietic stem cell transplantation (HSCT). The efficacy of DLI is, however, limited by the occurrence of graft-versus-host disease (GvHD). In three different clinical trials, we showed that the infusion of lymphocytes transduced by a retroviral vector expressing a suicide gene (HSV-TK) and a surface marker (ΔLNGFR) allows to control GvHD in 100% of cases, while preserving anti-tumor and antiviral activities. In >40 patients treated with TK-cells in the context of HLA identical and haplo-identical HSCT, we observed consistent expansion (up to 40% of circulating cells) and long-term persistence (>10 years) of transduced cells. No acute or chronic adverse or toxic effect due to the gene transfer procedure was observed in these patients, who were treated with a total of >1011 cells generated by >60 independent transductions. Analysis of vector integration sites was preformed by LM-PCR on lymphocytes obtained up to 10 years after treatment from 4 patients, and selected for ΔLNGFR expression. 60% of the sequences met our validity criteria and were unambiguously mapped onto the human genome by Ensembl BLAST analysis. Transduced T-cells were highly polyclonal, with vector integrations occurring preferentially within genes (52%), and particularly within the first intron (25%). Quantitative PCR analysis of selected integrations was performed to follow the dynamics of individual T-cell clones during time. Microchip analysis of the gene expression profile showed that <200 out of >22,000 genes (0.9%) were differentially expressed in ΔLNGFR+ vs. ΔLNGFR- T-cells from two different patients, suggesting that no significant perturbation was induced by retroviral transduction or HSV-TK/ΔLNGFR expression in human lymphocyte populations. Interestingly, these data also show the substantial biological identity of T-cell population generated by HSCT and those administered as DLI. Finally, the influence of proviral integration on the expression of targeted genes was studied in individual T-cell clones transduced in vitro or obtained ex vivo from treated patients by quantitative gene expression analysis.
The International Medical Prevention Registry on Venous Thromboembolism (IMPROVE): Venous Thromboembolism Prophylaxis Practices in Acutely Ill Medical Patients.
Authors: Tapson Victor F. , Decousus Herve , Bergmann Jean-Fran[ccedi]ois , Chong Beng H. , Froehlich James B. , Kakkar Ajay K. , Merli Geno J. , Monreal Manuel , Nakamura Mashio , Pavanello Riccardo , Pini Mario , Piovella Franco , Spyropoulos Alex C. , Turpie Alexander G.G. , Zotz Rainer B. , FitzGerald Gordon , Anderson Frederick A. ,   for the IMPROVE investigators
Published: October 2019

Abstract
Background Despite consensus group recommendations indicating that medical patients should receive appropriate venous thromboembolism (VTE) prophylaxis, prophylaxis practices remain poorly characterized. This analysis of IMPROVE, a prospective study of acutely ill medical patients, describes in-hospital prophylaxis practices prior to the publication of updated VTE prevention guidelines by the American College of Chest Physicians.
Methods Patient recruitment began in July 2002. Patients ≥18 years old, and hospitalized for ≥3 days with an acute medical illness are enrolled consecutively. Exclusion criteria are: therapeutic antithrombotics/thrombolytics at admission; major surgery or trauma during 3 months prior to admission; and VTE treatment begun within 24 hours of admission.
Results Data were from 4315 patients (32% from USA) enrolled up to 30 June 2004 in 37 hospitals in 11 countries (76% with 3-month follow-up data). Patients are 50% female, median (IQR) age 69 (50–80) years, median length of hospital stay 8 (5–14) days, median weight 68 (58–80) kg, and 40% were immobile for ≥3 days (median length of immobility 7 [4–14] days, including immobility immediately prior to admission). In-hospital VTE prophylaxis was received by 41% of patients (Table 1). Of patients with no risk factors (44%), one risk factor (40%), or ≥2 risk factors (16%), 25%, 49%, and 67% received prophylaxis, respectively. 12% of IMPROVE patients would have been eligible for inclusion in the MEDENOX study. Of these, only 52% received prophylaxis in hospital. Prophylaxis was provided to 6% of patients during the 3-month follow-up period, and continued in 11% of patients after discharge.
Conclusions Only 41% of IMPROVE patients received VTE prophylaxis, with considerable variation in types and regimens of prophylaxis used. While MEDENOX showed the benefits of VTE prophylaxis (enoxaparin 40 mg) in acutely ill medical patients, only half of MEDENOX-eligible patients received prophylaxis.
Table 1. Use of in-hospital VTE prophylaxis (N=4315)
VTE prophylaxis Patients receiving VTE prophylaxis, % ROW, rest of world; *Excluding elastic stockings and aspirin ≥1 type of VTE prophylaxis* 41 LMWH - USA (Q12h, Qd) 7 (5, 1) LMWH- ROW (Q12h, Qd) 31 (29, 2) UFH - USA (Q12h, Q8h) 28 (15, 11) UFH - ROW (Q12h, Q8h) 6 (5, 0) Intermittent pneumatic compression (USA, ROW) 6 (19, 0) Aspirin (USA, ROW) 4 (7, 3) Elastic stockings (USA, ROW) 6 (3, 8)
Safety and Efficacy of Alfimeprase for Restoring Function in Occluded Central Venous Catheters: Interim Results of a Phase 2, Multicenter, Randomized, Double-Blind Study (NuCath).
Authors: Deitcher Steven R. , Moll Stephan , Homesley Howard D. , Bertoli Luigi , Kenyon Peter , Swischuk Peter , Pena Luis 
Published: October 2019

Abstract
Alfimeprase, a recombinantly produced 22.6 kilodalton metalloproteinase, is a genetically modified variant of fibrolase. Alfimeprase, like fibrolase, proteolytically cleaves both the alpha and beta chains of fibrin(ogen) independent of plasminogen activation to plasmin and directly dissolves thrombi. Readily abundant alpha-2 macroglobulin acts as an endogenous circulating alfimeprase inactivator, forming a covalent, irreversible bond with alfimeprase which results in an inactive complex. Based on the direct thrombolytic effect of alfimeprase, rapid activity was hypothesized. We performed a Phase 2, randomized, double-blind, controlled, dose-ranging study to compare the safety and efficacy of one or two instillations of three intraluminal doses of alfimeprase (0.3 mg; 1.0 mg; and 3.0 mg) and Cathflo® Activase® (Genentech) 2.0 mg in reestablishing patency to occluded central venous access devices (CVADs). This report describes an interim analysis of 48 patients with CVAD withdrawal occlusion who were enrolled and randomized to treatment. Catheter patency was assessed at 5, 15, 30, and 120 minutes after each dose of study drug. Adverse events including bleeding events were assessed for a 30-day period after exposure to study drug. Cumulative patency rates are shown in the table. All study arms had similar patency rates at 120 minutes after the first dose. The alfimeprase 3.0 mg dose produced the highest patency rate at 120 minutes after the second dose. All three alfimeprase doses were more effective than Cathflo® Activase® during the first 30 minutes of treatment. The alfimeprase 1.0 mg and 3.0 mg doses resulted in ≥ 50% patency restoration rates at 15 minutes compared to 0% for Cathflo® Activase®. No intracranial hemorrhage, major hemorrhagic, or embolic events were reported in any treated patients at 5 days. Efficacy and safety results of this study support further evaluation of alfimeprase doses ranging from 1.0 mg to 3.0 mg for treatment of occluded CVADs.
Cumulative Patency Rate (%)
Alfimeprase 0.3 Alfimeprase 1 Alfimeprase 3 Cathflo Activase N=13 N=14 N=9 N=12 D=dose; min=minutes after dose Baseline 0 0 0 0 D 1; 5 min 15 14 44 0 D 1; 15 min 15 50 57 0 D 1; 30 min 30 50 57 25 D 1; 120 min 46 50 57 50 D 2; 5 min 46 50 57 58 D 2; 15 min 46 50 57 67 D 2; 30 min 46 50 57 67 D 2; 120 min 46 50 78 67
Prognostic Relevance of Lipoprotein Lipase (LPL) Expression in B-CLL.
Authors: Heintel Daniel , Kienle Dirk , Martin Bilban , Shehata Medhat , Kroeber Alexander , Mitteregger Dieter , Porpaczy Edith , Kainz Birgit , Auer Michael , Gleiss Andreas , Dohner Hartmut , Gaiger Alexander , Wagner Oswald , Stilgenbauer Stephan , Jaeger Ulrich 
Published: October 2019

Abstract
We investigated the pattern of lipoprotein lipase (LPL) expression in B-cell chronic lymphocytic leukemia (B-CLL) and assessed its prognostic and potential functional relevance. LPL mRNA levels in peripheral blood mononuclear cells (PBMNC) from 55 CLL patients differed by 2 orders of magnitude between cases with mutated (N=28) or unmutated (N=27) IGVH (median: 1.62 vs. 100.43 compared to normal PBMNC). LPL expression correlated strongly with IGVH mutational status (R=0.78; P<0.0001). High LPL expression (≥ 10) predicted unmutated IGVH status with an odds ratio of 128.71 (P < 0.0001) and discriminated between mutated and unmutated cases in 51 of 55 patients (93%). In an extended cohort of 104 patients, a trend for LPL as a prognostic factor regarding overall survival was found: While the median overall survival (OS) in patients with LPL expression of more than 10 was 105 months, the median OS was not reached in the low LPL group. Importantly, high LPL expression (≥ 10) was significantly associated with a short treatment-free survival (median 40 vs. 96 months, P = 0.001). By indirect immunofluorescence, LPL was detected in the cytoplasm of leukemic B-CLL cells and showed intense, large granular and punctate distribution. Fine granular and diffuse staining was observed in monocytes but no immunoreactivity was detected in T- cells. In order to compare the LPL protein expression unmutated (LPL mRNA high) and mutated (LPL mRNA low) samples from 10 patients (5 mutated and 5 unmutated) were prepared and stained under identical conditions.The intensity of intracellular immunoreactivity to LPL antibody in B-CLL cells was higher in each unmutated compared to the mutated samples. In order to investigate the association of LPL with other genes we performed gene expression profiling on 12 patient samples selected for high or low LPL yielding 251 differentially expressed genes: 171 genes associated with LPL only (group 1), 58 genes with LPL and mutational status (group 2) and 22 genes with mutational status only (group 3). While group 2 and 3 contained a number of genes previously described by Rosenwald and Klein*, a considerable number of genes associated with lipid metabolism were clustered in group 1, indicating a functional role of LPL in B-CLL cells. Our data establish LPL as a prognostic marker and suggest functional consequences of LPL overexpression in patients with B-CLL.
*J Exp Med 2001
PROphylaxis for ThromboEmbolism in Critical Care Trial (PROTECT): A Pilot Study.
Authors: McDonald Ellen , Poirier Germain , Hebert Paul , Pagliarello Joe , Rocker Graeme , Langevin Stephane , LeBlanc Francois , Mehta Geeta , Skrobik Yoanna , Fowler Robert , Granton John , Freitag Andreas , Jones Graham , Cooper Jamie , Meade Maureen , Guyatt Gordon , Anderson David , Geerts Bill , Crowther Mark , Zytaruk Nicole , Griffith Lauren , Cook Deborah ,   For the PROTECT Investigators and the Canadian Critical Care Trials Group
Published: October 2019

Abstract
Rationale: No published randomized trials have compared low molecular weight heparin (LMWH) to unfractionated heparin (UFH) for thromboprophylaxis in medical-surgical ICU patients.
Objectives: A) Our Feasibility Objectives were to assess: 1) timely enrollment and complete, blinded study drug administration, 2) LMWH bioaccumulation in renal insufficiency, 3) twice weekly leg ultrasounds, and 4) recruitment rates for a future trial. B) Our Clinical Objectives were to assess 1) the effectiveness of LMWH vs UFH by estimating DVT, and pulmonary embolism (PE) rates, and 2) the safety of LMWH vs UFH regarding bleeding, thrombocytopenia and HIT.
Methods: In this prospective randomized stratified concealed blinded multicenter trial, we included patients ≥18 with expected ICU stay ≥72h. We excluded trauma, orthopaedic, cardiac, or neurosurgery, severe hypertension, DVT, PE or hemorrhage within 3 mos, INR >2ULN, PTT >2ULN, platelets <100 x109/L, creatinine clearance <30ml/min, therapeutic anticoagulation, >2 doses of heparin, and palliative care patients. By centralized telephone randomization, we allocated patients to dalteparin 5,000 IU QD SC or UFH 5,000 IU BID SC. All clinicians caring for patients were blinded except the pharmacist. Bilateral leg compression ultrasounds were performed within 48h of ICU admission, twice weekly, on suspicion of DVT, and 7 days post ICU discharge.
Results: A) Feasibility Objectives: 1) Timely, complete study drug administration occurred for >99% of scheduled doses; every dose was blinded. 2) No LWMH bioaccumulation was observed as measured by anti-Xa levels when creatinine clearance decreased to <30 ml/min. 3) Ultrasounds were performed without exception. 4) Estimated recruitment for a future trial is 4–5 patients/month/center. B) Clinical Objectives: 1) Effectiveness: Among 128 patients, 11 (8.6 %) had DVT, 2 (1.6 %) had PE. 2) Safety: 6 (4.7 %) had clinically important bleeds, 23 (18 %) had minor bleeds, 4 (3.1%) developed a platelet count <50 x109/L, and 2 (0.8%) had HIT diagnosed by serotonin release assay.
Conclusions: DVT rates remain high in medical-surgical ICU patients despite universal heparin thromboprophylaxis. This pilot study suggests that a randomized clinical trial comparing LMWH vs UFH feasible. An adequately powered trial is needed to achieve the clinical objectives.
Argatroban Inhibits Human Factor Xa at Therapeutic Drug Levels: An Explanation for Its Prolongation of the INR.
Authors: Warkentin Theodore E. , Greinacher Andreas , Craven Sharon , Dewar Lori , Sheppard Jo-Ann I. , Ofosu Frederick A. 
Published: October 2019

Abstract
Three direct thrombin inhibitors (DTIs) differ in their ability to increase the International Normalized Ratio (INR) for a given prolongation of the activated partial thromboplastin time (APTT), as follows: argatroban > bivalirudin > lepirudin. This phenomenon is clinically-relevant, as it can complicate the transition to coumarin therapy (a risk factor for venous limb ischemia in patients with heparin-induced thrombocytopenia and deep-vein thrombosis, i.e., a clinical situation in which a DTI may be given). Since this observed order of INR prolongation by DTIs is the reverse of that expected based on their known relative affinities for human thrombin (see Table), we tested whether these DTIs (as well as the small-molecule thrombin inhibitor, D-Phe-Pro-ArgCH2Cl [PPACK]) also inhibit free and/or prothrombinase-bound factor Xa (FXa). Prothrombinase-bound FXa was assembled on rabbit brain phospholipids supplemented with human plasma factor X-depleted plasma, to which human FXa (0.5 nM, final) was added. We found that argatroban inhibited free human FXa, i.e., FXa in the fluid-phase (IC50 = 5 μM), whereas it inhibited bovine FXa much less effectively (IC50 = 110 μM). In contrast, only supratherapeutic concentrations of bivalirudin inhibited human and bovine FXa, whereas lepirudin had no effect (see Table). Neither therapeutic nor supratherapeutic concentrations of the three DTIs inhibited prothrombinase-bound FXa.
Comparison of DTIs on INR, APTT, IIa, and Inhibition of Human and Bovine Fxa
PPACK Argatroban Bivalirudin Lepirudin * published values Effect on INR N.D. Greatest Intermediate Least [DTI] doubling APTT N.D. 1 μM 0.25 μM 0.06 μM Ki for human thrombin* ~40 nM ~2 nM ~0.0001 nM IC50 human FXa 3 μM 5 μM 1,700 μM >10,000 μM IC50 bovine FXa 5 μM 110 μM 2,900 μM >10,000 μM
CONCLUSION: Compared with the other two DTIs, argatroban effectively inhibits human FXa at concentrations (IC50 = 5 μM) within the order of magnitude of the usual therapeutic drug levels (about 1 to 2 μM). Inhibition of free FXa is a plausible reason why argatroban prolongs the INR more effectively than the other two DTIs.
Development of Prothrombinase Induced Clotting Time (Pefakit ®, PiCT) Assay and its Validation in the Monitoring of Anti-Xa and Anti-FIIa Drugs.
Authors: Hoppensteadt Debra A. , Cunanan Josephine , Walenga Jeanine M. , Fareed Jawed 
Published: October 2019

Abstract
Unfractionated heparin (UFH), low molecular weight heparin (LMWH), fondaparinux, danaparoid, direct and indirect (via a cofactor) acting anti-FIIa and anti-Xa anticoagulant drugs are being used in patients for the prophylaxis and treatment of thrombosis. The aPTT does not reliably measure the effects of these drugs. Therefore, there is no simple/reliable method for monitoring patients administered these drugs, particularly those with renal insufficiency and obesity. The newly developed Prothrombinase Induced Clotting Time (PiCT) assay (Pefakit®; Pentapharm, Ltd.; Basel, Switzerland) was developed for monitoring heparins. The PiCT is based on the activation of plasma coagulation by a defined amount of activated FXa, phospholipid and Russell’s viper venom (RVV). The prothrombinase complex formed following recalcification is independent of endogenous thrombin mediated FV activation because the RVV directly activates FV. In this study, the PiCT was used to determine the relative anticoagulant actions of UFH, LMWH, anti-FIIa and anti-FXa drugs. Drugs were supplemented to normal human pooled plasma in a concentration range of 10- 0.08 μg/ml. Plasma samples were analyzed using the PiCT, aPTT, Heptest, thrombin time (TT), ecarin clotting time (ECT), chromogenic anti-FIIa, chromogenic anti-FXa and fibrinopeptide A (FPA) generation assays. In addition, plasma samples from patients treated with these drugs were also tested. The UFH, LMWH, anti-FIIa drugs and the indirect acting anti-FXa drugs produced a concentration-dependent prolongation of the PiCT. Different responses were obtained for each drug. The anti-FIIa drugs gave the strongest dose-response response followed by UFH, LMWH and fondaparinux, in that order. The direct acting anti-FXa agents showed a weak response in the PiCT. Compared to the other assays, the PiCT response showed better sensitivity, reproducibility and linearity over a broader dose range than the aPTT, Heptest, TT, ECT, anti-FIIa and anti-FXa assays. Samples from patients treated with enoxaparin (0.5 mg/kg IV) gave peak concentrations of 0.92± 0.11 U/ml by the PiCT, compared to 0.95± 0.24 U/ml by the anti-FXa assay. Samples from patients treated with bivalirudin, lepirudin or argatroban gave comparable values by the PiCT, aPTT and ECT. The relative prolongation of the PiCT was proportional to the inhibition of thrombin generation as measured by FPA. The PiCT is as simple to perform as the aPTT and can be adapted to any instrument capable of detecting a fibrin clot. Unlike other clot-based tests, the PiCT is sensitive to most old and new anticoagulant drugs and the results are linear through both the therapeutic and interventional dosing ranges. The PiCT may offer the first available single assay that can be universally used for monitoring most of the new anticoagulant drugs regardless of their mechanism of action.
Incidence and Disease Pattern of Venous Thrombembolism after Radical Prostatectomy.
Authors: Beyer Jan , Wessela Sven , Hakenberg Oliver , Wirth Manfred , Schellong Sebastian 
Published: October 2019

Abstract
Background: Little is known about the time of development and distribution pattern of venous thrombembolism (VTE) after radical prostatectomy (RPE) for malignancy.
Patients and Methods: 415 consecutive patients undergoing standard RPE were prospectively assessed regarding the incidence and distribution pattern of VTE, the hemodynamic relevance of pelvic lymphoceles (LC) and other possible risk factors for VTE. All patients underwent RPE and received heparin prophylaxis until discharge, followed by acetylsalicylic acid 100 mg daily. On day -1, 8 and 21 complete compression ultrasound, measurement of pelvic vein flow and pelvic ultrasound were performed.
Results: VTE was found in 17,3% of patients. Most thrombi were limited to calf muscle veins (55,5%). Isolated calf deep vein thrombosis (DVT) was found in 22,2% and proximal DVT in 12,5%. On day 8 and 21 the following incidences of VTE were found: calf muscle vein thrombosis 4.8 and 4.8%; isolated calf vein thrombosis 1.7% and 2.2%; proximal DVT 0.5% and 1.7%; pulmonary embolism 0.5% and 0.5%, respectively. Multivariate analysis revealed surgery time (OR 1.26), number of blood tranfusion (OR 1.73) and venous flow reduction (OR 2.83) as risk factors for the development of VTE. 33.3% of the patients developed a LC postoperatively; of these 5.3% showed a reduction of pelvic vein flow. A venous flow reduction was only seen with LC.
Conclusion: The incidence and distribution pattern of VTE after RPE is comparable to major abdominal surgery for malignancy. The development of LC is a significant risk factor for VTE, when pelvic vein flow is impaired. Since the majority of proximal VTE developed between day 8 and day 21 when the patient is already discharged, the concept of a prolonged heparin prophylaxis after RPE should be evaluated prospectively.
Azacitidine (Vidaza®) Treatment Response Assessed Using Three Alternative Dosing Schedules in Patients with Myelodysplastic Syndromes (MDS).
Authors: Lyons Roger , Cosgriff Thomas , Modi Sanjiv , Lintz Linda , Beach C.L. , Backstrom Jay T. 
Published: October 2019

Abstract
Previous CALGB trials published by Silverman et al (JCO2002;20:2429) have shown azacitidine to be efficacious and well tolerated in the treatment of MDS at a dosing schedule of 75 mg/m2/day x 7 days every 28 days. The objective of this phase II, multicenter, randomized, open-label trial was to study treatment response in patients with MDS with 3 alternative, subcutaneous azacitidine dosing regimens, each eliminating the need for weekend injections. The 3 alternative dosing schedules, repeated in 28-day cycles, were AZA 5-2-2 (75 mg/m2/day x 5 days, followed by 2 days of no treatment, followed by 75 mg/m2/day x 2 days), AZA 5-2-5 (50 mg/m2/day x 5 days, followed by 2 days of no treatment, followed by 50 mg/m2/day x 5 days), and AZA 5 (75 mg/m2/day x 5 days). Eligible MDS patients must have had a life expectancy of ≥ 7 months and an ECOG performance status grade of 0–3, with a FAB classification of RA, RARS, RAEB, RAEB-T, or CMML. RA and RARS patients must also have had either hemoglobin < 110 g/L with transfusion requirements, a platelet count < 100 x 109/L, or an ANC < 1.5 x 109/L. Patients experiencing International Working Group response criteria for MDS (Blood2000;96:3671) defined as complete remission, partial remission, stable disease, or hematologic improvement (major or minor) after 6 cycles of treatment were eligible to receive an additional 12 cycles. A total of 44 patients have been randomized to date with 22, 14, and 8 receiving either AZA 5-2-2, AZA 5-2-5, or AZA 5, respectively. As the AZA 5 day dosing schedule was added later to the protocol by amendment, no patients in this arm are currently evaluable for response. Most patients are male (64%) and elderly with a median age of 74 years. Most patients have RA, using either FAB (45%) or WHO (36%) definitions; another 27% and 16% have RAEB or RAEB-1, respectively. As azacitidine is incorporated into DNA/RNA through successive cell cycles, patients often require a minimum of 4 to 6 cycles of azacitidine before responses are observed. Early results show that the efficacy and tolerability observed with the alternative dosing schedules are consistent with the 75 mg/m2/day x 7 days dosing results from previous studies.