bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Safety of Co-Administered Low-Molecular-Weight Heparin and Continuous Epidural Analgesia after Major Orthopedic Surgery: Assessment of a Standardized Management Protocol.
Authors: Wang Jenny , Douketis James , Wagler Meghan , Kinnon Krys , Crowther Mark 
Published: October 2019

Abstract
Background: In patients who have had major orthopedic surgery, low-molecular-weight heparin (LMWH) has established efficacy in preventing venous thromboembolism (VTE) after surgery, and continuous epidural analgesia (CEA) provides better pain control than parenteral analgesia and may reduce the risk of cardiorespiratory complications. However, there is concern about the co-administration of LMWH and CEA after surgery because of an increased risk of epidural hematoma. No study, to our knowledge, has investigated the safety of a standardized treatment regimen that allows co-administered LMWH and CEA after surgery.
Patients and Methods: We assessed the safety of a standardized management protocol in 970 patients who were scheduled to undergo knee or hip replacement surgery and who received post-operative thromboprophylaxis with LMWH and pain control with CEA. All patients received intra-operative spinal (regional) anesthesia, with an indwelling epidural catheter left in place after surgery to administer epidural analgesia with an infusion of fentanyl and bupivacaine. Subcutaneous LMWH (enoxaparin 30 mg twice-daily or dalteparin 5000 IU daily) was started on the morning after surgery and continued for at least 5 days. CEA was administered for 2–3 days after surgery, with the epidural catheter removal occurring during the trough anticoagulant effect of the LMWH, 10 hours and 22 hours after the preceding dose of enoxaparin and dalteparin, respectively. In patients who were to receive warfarin, this treatment was initiated, with a 2.5 mg dose, on the evening prior to the scheduled removal of the epidural catheter. Drugs that are known to affect hemostasis were avoided during CEA, and patients at increased risk for bleeding were not given LMWH. During the period of hospitalization, we calculated the incidence and associated 95% confidence interval (CI) for the following outcomes: epidural hematoma; other bleeding (major and minor); VTE, myocardial infarction, stroke, death.
Results: 970 patients (mean age: 68.2 years; mean weight: 81.4 kg; 63% female) who underwent knee replacement (527 patients) or hip replacement (443 patients) received co-administered LMWH and CEA. The mean duration of LWMH therapy was 7.2±5.3 days. The mean duration (±standard deviation) of CEA was 1.9±0.80 days, respectively. Warfarin was initiated in 758 (78%) patients. There were no episodes of symptomatic epidural hematoma (0%; 95% CI: 0–0.38). During the in-hospital follow-up period, there were 13 episodes of VTE (1.3%; 95% CI: 0.71–2.3), 47 bleeds (4.7%; 95% CI: 3.5–6.3); 4 myocardial infarcts (0.41%; 95% CI: 0.12–1.1), and 4 deaths (0.41%; 95% CI: 0.12–1.1).
Conclusion: Co-administered LMWH and CEA appears safe after major orthopedic surgery when using a standardized management protocol.
Initial Outpatient Treatment of Venous Thromboembolism with Fondaparinux (Arixtra®): The Matisse Trials.
Authors: Buller Harry R. ,   the MATISSE Investigators
Published: October 2019

Abstract
Background: The MATISSE trials showed that a single dose regimen of fondaparinux, a synthetic selective factor Xa inhibitor, was at least as effective and as safe as standard therapies in the treatment of venous thromboembolism (VTE). In these trials, outpatient treatment of fondaparinux was encouraged but left at the investigator’s discretion. We analyzed the data in patients who received fondaparinux on an outpatient basis.
Methods: Fondaparinux was administered at a once-daily subcutaneous dose of 7.5 mg (5.0 mg and 10.0 mg in patients <50 kg and >100 kg, respectively). In the MATISSE-DVT trial, fondaparinux was compared with twice-daily subcutaneous enoxaparin (1 mg/kg) in patients with deep-vein thrombosis (DVT). In the MATISSE-PE trial, it was compared with adjusted-dose intravenous unfractionated heparin (UFH) in patients with pulmonary embolism (PE). Outpatient treatment of DVT with enoxaparin was possible whereas outpatient treatment of PE with UFH was not feasible. All drugs were given for at least 5 days and until anticoagulation with oral anticoagulants was therapeutic. The primary efficacy and safety outcomes were recurrent VTE during 3 months’ follow-up and major bleeding (MB) and death during the initial treatment period.
Results: In MATISSE-DVT, 31.4% and 33.8% of the fondaparinux- and enoxaparin-treated patients, respectively, received therapy on an outpatient basis. In MATISSE-PE, 14.3% of the patients received fondaparinux on an outpatient basis, compared with none in the UFH group. In both MATISSE-DVT and -PE, efficacy and safety data from the patients who received fondaparinux on an outpatient basis were similar to those from the total population (Tables). The rates of recurrent VTE and MB in fondaparinux outpatients were similar to those in enoxaparin outpatients or UFH inpatients.
Conclusion: Outpatient initial treatment of both DVT and PE with once-daily fondaparinux is feasible, effective and safe.
MATISSE DVT Enoxaparin Fondaparinux All patients Outpatients All patients Outpatients *As treated patients n 1107 374 (33.8%) 1098 345 (31.4%) Age, yr (mean±SD) 61±17 60±16 61±17 58±17 Male/female 578/529 201/173 581/517 197/148 Hospital discharge, days (mean±SD) 7.0±6.2 1.8±1.9 7.6±7.7 1.6±1.7 ≥2 VTE risk factors, n (%) 283 (25.6) 122 (32.6) 293 (26.7) 97 (28.1) VTE, n (%) 45 (4.1) 16 (4.3) 43 (3.9) 7 (2.0) MB*, n (%) 13 (1.2) 3 (0.8) 12 (1.1) 5 (1.5) MATISSE-PE UFH Fondaparinux All patients Outpatients *As treated patients n 1110 1103 158 (14.3%) Age, yr (mean±SD) 62±17 63±16 57±16 Male/female 477/633 501/601 82/76 Hospital discharge, days (mean±SD) 10.2±6.8 9.7±7.7 4.4±2.2 ≥2 VTE risk factors, n (%) 260 (23.4) 241 (21.8) 35 (22.2) VTE, n (%) 56 (5.0) 42 (3.8) 5 (3.2) MB*, n (%) 12 (1.1) 14 (1.3) 0 (0)
BCR/ABL Expression Increases the Formation of Chromosomal Translocations after DNA Damage.
Authors: Dierov Jamil , Padilla-Nash Hesed , Ried Thomas , Carroll Martin 
Published: October 2019

Abstract
BCR/ABL is the protein product of the t(9;22) translocation and is the cause of the hyperproliferation associated with chronic phase chronic myeloid leukemia (CML). However, whether BCR/ABL induces genomic instability leading to blast crisis is controversial. We have previously demonstrated that BCR/ABL translocates to the nucleus after genotoxic damage and associates with the DNA damage sensor, ataxia telangiectasia and rad 3 related (ATR) protein, disrupting the sensing and repair of DNA double strand breaks. Here, we have asked if BCR/ABL expression leads to permanent changes in the DNA after genotoxic stress. For these experiments we have studied the hematopoietic cell line, Ba/F3pTetOn p210, which expresses p210 BCR/ABL after incubation in doxycycline. Cells were incubated in low doses of etoposide for two hours and then allowed to recover for 48 hours in the absence of further DNA damage. Induction of apoptosis in these conditions was consistently less than 5% as demonstrated by annexin V staining of cells. Cells were examined for alterations in the chromosomes using Giemsa banding and spectral karyotyping (SKY). Cells growing in IL3 showed low levels of DNA damage with a few broken chromosomes present in metaphase spreads and an average of 0.5 new chromosomal translocations per cell as revealed by SKY analysis. In contrast, when cells expressing BCR/ABL were treated with the same conditions, a marked number of genetic abnormalities were seen. 75% of cells showed abnormal chromosome forms with ring chromosomes, triradial forms and other abnormalitites. Analysis of BCR/ABL expressing cells by SKY analysis showed frequent abnormalities, averaging at least 6 new translocations per cell. Several cells had greater than 10 translocations present including multiple complex translocations involving more than two chromosomes. The majority of abnormalities consisted of unbalanced translocations. This data demonstrates that BCR/ABL alters the cellular response to DNA damage leading to an increase in chromosomal translocations in cells expressing the oncogene and suggests that BCR/ABL itself is directly responsible for the genomic instability leading to CML blast crisis.
Differential Phosphorylation of FANCA and FANCG Determines Response of Fanconi Anemia Core Complex to DNA Damage and S Phase.
Authors: Mi Jun , Tomashevski Andrei , Kupfer Gary M. 
Published: October 2019

Abstract
Fanconi anemia (FA) is a genetic disease marked by bone marrow failure, congenital defects, and cancer. In spite of the identification of at least 8 genes, the biochemistry of the disease and its normal pathway in the cell remains elusive. The FA core complex is composed of at least 5 proteins, 2 of which, FANCA and FANCG, we have shown to be phosphorylated. In these studies, we show that both FANCA and FANCG are phosphorylated in response to DNA damage. In the case of FANCG, we have mapped the site of this phosphorylation to serine 7, using a phosphoserine 7 FANCG antiserum. Because of the link of FA function and the FA core complex-dependent monoubiquitination that occurs both as a result of DNA damage as well as at S phase, we also examined if phosphorylation occurred at S phase as well. While FANCG serine 7 phosphorylation occurs both at S phase and after DNA damage (similar to FANCD2 monoubiquitination), FANCA phosphorylation occurs only after DNA damage. Recent data have implicated the kinase ATR as important in the pathway. In order to assess whether a downstream target of ATR is differentially phosphorylated in FA cells, we tested the phosphorylation status of chk1 in FA-A mutant and corrected cells. Chk1 kinase is phosphorylated at serine 318 in response to DNA damage only in corrected cells but not mutant FA cells, while signaling through chk2 kinase is unaffected. These data suggest the importance of phosphorylation in the FA pathway in the regulation of both cellular responses to DNA damage as well as engagement of the cell cycle.
Primary Treatment of Waldenstrom’s Macroglobulinemia (WM) with Dexamethasone, Rituximab and Cyclophosphamide.
Authors: Dimopoulos Meletios A. , Kyrtsonis Maria Christina , Tsatalas Constantinos , Kokkini Garyfallia , Repoussis Panagiotis , Zomas Athanasios , Symeonidis Argiris , Michali Evridiki , Anagnostopoulos Athanasios , Anagnostopoulos Nikolaos , Economopoulos Theophanis , Pangalis Gerasimos 
Published: October 2019

Abstract
Introduction: The three main choices for the primary treatment of WM are alkylating agents, nucleoside analogues and rituximab. There is evidence that combinations of nucleoside analogues and rituximab are highly active. However, when such regimens are applied, there are concerns regarding myelosuppression, immunosuppression and impact on stem cell collection. We designed a phase II study which included previously untreated WM patients (with clearly defined criteria for initiation of treatment) who were treated with a combination of dexamethasone, rituximab and cyclophosphamide.
Patients and methods: Patients with WM received primary treatment with dexamethasone 20 mg i.v. followed by rituximab 375mg/m2 i.v. on day 1 and cyclophosphamide 100mg/m2 p.o. bid days 1–5 (total dose 1000 mg/m2). This regimen was repeated every 21 days for 6 courses.
Results: Since July 2002, 34 patients have been enrolled in this multicenter study. Patients characteristics include: median age of 75 years (range 53 to 86 years), 59% male, 25% B symptoms, 25% splenomegaly, 35% lymphadenopathy, 25%hyperviscosity syndrome, 59% hemoglobin < 10 g/dl, 37% serum monoclonal protein ≥ 4 g/dl, 39% serum albumin < 3.5 g/dl, 54% serum β2 microglobulin > 4 mg/dl. So far, on an intent-to-treat basis, 78% of patients achieved at least 50% reduction of serum monoclonal protein concentration. The median time to response was 3.6 months (range 0.7 months to 5.4 months). With a mean follow-up of 18 months, 90% of patients are progression-free. With the exception of one patient who died of interstitial pneumonia, the DRC regimen was well tolerated. 15% of patients developed grade 3, 4 neutropenia. Mild or moderate effects related to rituximab infusion such as fever, rigors and headache were seen in 20% of patients.
Conclusions: We administered a regimen which combined rituximab with cyclophosphamide, a non-stem cell toxic alkylating agent. The DRC regimen is an active and well tolerated primary treatment for symptomatic patients with WM. Patient accrual and evaluation is ongoing.
Immunotherapy of Chronic Lymphocytic Leukemia using CD40L and IL2 Expressing Autologous Tumor Cells.
Authors: Biagi Ettore , Popat Uday , Raphael Rousseau , Eric Yvon , Giampietro Dotti , Lawrence Rice , Andreef Michael , Brenner Malcolm 
Published: October 2019

Abstract
Transgenic human CD40 ligand (hCD40L) activates B-Chronic Lymphocytic Leukemia (B-CLL) cells by CD40 stimulation and may thereby enhance their capacity to present tumor antigens. Pre-clinical models show that co-expression of IL-2 further potentiates the immunogenicity of CD40L-expressing tumor cells. To discover whether these promising pre-clinical data could usefully be applied to patients with B-CLL, we used adenovectors to prepare hIL2- and hCD40L-expressing autologous tumor vaccines. Within these vaccines, a mean of 92% B-CLL cells were CD40L positive (versus <1% pre-treatment), and costimulatory molecules were also upregulated on the tumor cells (CD80 from 6% to 74% and CD86 from 16% to 73% positive cells). The mean secretion of IL-2 (measured by ELISA at 72 hours after transduction) was 1,780 pg/ml/10E6 cells (range = 175–400). To date, 8 patients have received between 5 and 8 subcutaneous injections in a modified Phase I design, in which the dose of IL-2 secreting cells was fixed at 2x10E7 per injection, while the dose of CD40L-expressing leukemia cells was escalated from 2x10E5 (level 1) to 2x10E7 (level 3) per injection. 12 additional patients will receive this final dose combination. Of those treated, two patients were in Rai stage 4; 2 in stage 3; 2 in stage 2; and 2 in stage 1. There were no adverse events from any injection in the patients. Injection-site biopsies revealed a modest infiltration of CD3+ cells, and the vaccine also had systemic immunomodulatory effects. Total T-cell numbers were significantly increased in only 2 patients, but an anti-B-CLL immune response was detected in 5/6 individuals currently analyzed. Using ELISPOT assays with autologous B-CLL cells as stimulators, and allogeneic B-CLL cells as controls, we found a rise in IFN-gamma spot-forming T-cells (<1/100,000 pre to >1/1500 post vaccination) and of Granzyme-B expressing T-cells (<1/100,000 pre to >1/2500 post). This reactivity is mediated at least in part by CD8-positive T cells with specificity to survivin, a known B-CLL associated tumor antigen. Although these immunologic effects have as yet been accompanied by only transient disease responses, our continuing accrual of additional patients who will receive immunomodulatory doses of vaccine should indicate whether the approach could contribute to the management of early or advanced B-CLL.
NIPA Targets Nuclear Cyclin B1 in a Cell-Cycle Dependent Manner.
Authors: Bassermann Florian C. , von Klitzing Christine , Kluempen Silvia , Bai Ren-Yuan , Ouyang Tao , Morris Stephan W. , Peschel Christian , Duyster Justus 
Published: October 2019

Abstract
Ubiquitin-mediated destruction of regulatory proteins marks the vital means of controlling cell cycle progresssion. The E3 ubiquitin-ligases are prominent in this process, as they allow the transfer of ubiquitin to the target protein and mediate substrate binding specificity. Recently, a new class of E3 ligases referred to as SCF complexes has been identified that consists of four subunits:SKP1, Cul1, Roc1 and an F-box protein, the latter of which determines substrate specifity.
We previously reported the cloning of NIPA (nuclear interaction partner of ALK) in complex with constitutively-active oncogenic fusions of ALK, which contribute to the development of certain lymphomas and sarcomas. Subsequently we characterized NIPA as a human F-box protein that determines a novel SCF complex (SCFNIPA) whose cell cycle regulated activity is restricted to interphase to allow for substrate expression at G2/M and mitosis. Phosphorylation of NIPA in late S-phase was found to be the underlying mechanism of SCFNIPA inactivation.
We have now identified the key mitotic regulator cyclin B1 to serve as the relevant substrate of the SCFNIPA complex. This targeting process is restricted to interphase and directed towards the nuclear pool of cyclin B1. Inactivation of NIPA by siRNAs results in nuclear accumulation of cyclin B1 in interphase and an elevation of cells in S-phase and mitosis. In contrast, expression of a phosphorylation deficient NIPA mutant that retains constitutive SCFNIPA activity throughout the cell cycle arrests cells at early prophase thus delaying mitotic entry. Both effects are likely attributable to either cyclin B1 accumulation in the case of NIPA inactivation by siRNA or untimely cyclin B degradation at G2/M upon expression of the constitutively active SCFNIPA complex. Cyclin B1 is physiologically kept cytoplasmic during interphase and premature nuclear accumulation has been associated with untimely mitotic entry, loss of checkpoint control and genomic instability. Our data provides a mechanism to inhibit premature nuclear accumulation of cyclin B1 in the mammalian cell cycle. NIPAs association with NPM-ALK of ALCL has been shown to be associated with NIPA phosphorylation and thus to the inactivation of the SCFNIPA complex. The mechanism described above may therefore provide a framework for understanding how this oncogene interferes with the physiologic regulation of cyclin B - a potential mechanism by which NPM-ALK transforms hematopoietic cells.
Adenoviral Mediated TGF-β1inhibition in a Murine Model of Myelofibrosis Can Prevent and Cure Bone Marrow Fibrosis Development.
Authors: Gastinne Thomas , Wagner-Ballon Orianne , Tulliez Micheline , Chagraoui Hedia , Vainchenker William , Giraudier Stephane 
Published: October 2019

Abstract
Myelofibrosis is a clinical feature of several hematopoietic disorders and is most prominent in idiopathic myelofibrosis. It is characterized by excessive deposits of extracellular matrix proteins which occur as a marrow microenvironment reactive response to cytokines released from the clonal malignant myeloproliferation. The observation that mice exposed to high systemic levels of thrombopoietin (TPO) invariably develop a myelofibrosis within a period of 4 to 8 weeks has allowed the demonstration of the crucial role of TGF-b1 released by hematopoietic cells in the promotion of myelofibrosis (Chagraoui et al Blood, 2002 ; 100 :3495). The aim of this study was to investigate whether TGF-b1 inhibition could directly inhibit the fibrosis development in a curative approach of this murine model.
An adenovirus encoding for TGF-b1 soluble receptor was constructed. SCID mice were infected with this adenovirus to measure the TGF-b1 inhibitory effect. We measure TGF-b1-Rs (TGF-b1 soluble receptor) concentration and TGF-b1 levels in 5 groups of mice injected with 0- 6.5 106- 6.5 107- 6.5 108-and 6.5 109 pfu/mice. We demonstrated that 6.5 108 pfu/mice was able to completely inhibit 10 fold increase of circulating TGF-b1 and that 1 microgram of soluble receptor was able to inhibit 1 ng of TGF-b1.
All subsequent experiments were performed by injecting 1 109 pfu/mice either shortly after transplantation (preventive) or after 30 days (curative). Briefly, 46 mice (2 individuals experiments n= 16 and n= 30) were transplanted with 5-FU treated syngeneic bone marrow cells transduced with a retrovirus encoding for murine TPO. Retroviral transduction efficiency varied from 67% to 87%. All mice developed a myeloproliferative syndrome characterized by an increase in platelet count, a leukocytosis, an increase in progenitor cells in the blood circulation and a 1000-fold increase in TPO level. TGF-b Rs (13 to 32 micrograms/ml) was detected in the blood of all mice injected with the TGF-b1-Rs Adeno. A dramatic decrease in TGF-b1 level was noticed in all these treated mice in comparison to the control groups. Histological analysis demonstrated that the two approaches (curative or preventive) were identical to abolish fibrosis development in bone marrow as well as in the spleen. However, all mice which received the TGF-b1-RS adenovirus developed a hepatitis leading to death 6 weeks to 3 months after infections.
In conclusion our approach support the major role of TGF-b1 in myelofibrosis development as previously demonstrated by our group using TGF-b1 KO mice. Moreover, the present results suggest that TGF-b1 inhibition could be a relevant approach to treat patients presenting a TGF-b1 mediated bone marrow fibrosis.
In Vitro Analysis of Myeloid Proliferation Caused by Oncogenic Kras.
Authors: Braun Benjamin S. , Gavin Joehleen , Tuveson David A. , Jacks Tyler E. 
Published: October 2019

Abstract
Somatic mutations in NRAS and KRAS2 are found in approximately one third of cases of myelodysplastic syndrome (MDS), myeloproliferative disorders (MPD) or acute myeloid leukemia (AML). Cases without RAS mutations frequently demonstrate genetic lesions which result in activation of Ras, such as FLT3 mutations, Bcr-Abl expression, or inactivation of the NF1 tumor suppressor. We have exploited a conditional oncogenic Kras allele that was generated by homologous recombination to directly address the effects of expressing a mutant K-RasG12D protein from the endogenous promoter in primary hematopoietic cells. We crossed this mutation into the Mx1-Cre strain, and induced Cre recombinase expression by administering polyinosinic-polycytidilic acid. These mice uniformly and rapidly develop a progressive MPD with leukocytosis and massive splenomegaly, and succumb after a median of 84 days. In vitro, myeloid cells expressing an oncogenic allele of Kras from the endogenous promoter demonstrate a dramatic hyperproliferative phenotype and hypersensitivity to hematopoietic growth factors. We have further characterized these findings, and have found that, despite the rapid accumulation of mature myeloid cells in vivo and in vitro, proliferation occurs mainly in immature, Mac1-negative progenitors. Therefore, myelopoiesis in Mx1-Cre, KrasG12D mice is analagous to, but more productive than that in normal mice. Kinetic data is most consistent with a major effect of K-rasG12D on cell cycle activity, with minimal effects on apoptosis. This system provides a novel and tractable platform for determining how oncogenic RAS alters myeloid homeostasis, for investigating how oncogenic RAS influences signaling networks in primary cells, and for preclinical testing of targeted therapeutics in primary cells.