bluuebunny/crossref_metadata_2025_split huggingface dataset:
Downregulating the Fibromodulin Gene by Short Interfering RNA Causes B-CLL Cell Death.
Authors: Choudhury Aniruddha , Derkow Katja , Mikaelsson Eva , Kokhaei Parviz , Österborg Anders , Rabbani Hodjattallah , Mellstedt Håkan 
Published: October 2019

Abstract
Development of targeted therapies against B-CLL is dependent on the identification of molecules that are essential for the proliferation and survival of the leukemic cells. One such molecule investigated by us as a putative leukemia-associated target is Fibromodulin (FIM). FIM is an extracellular matrix molecule belonging to the leucin-rich proteoglycan family. Our laboratory studies have verified that expression of FIM is upregulated in B-CLL cells. Moreover, this molecule is specifically overexpressed in B-CLL cells and not on normal peripheral blood mononuclear cells. Analysis of FIM expression on various other hematological tumor cells have revealed that with the exception of mantle cell lymphoma, FIM is not expressed in other hematological malignancies.
RNAi technology has recently emerged as a powerful method to specifically silence the expression of a gene. We have generated three siRNA against various segments of FIM and tested the effects of these siRNA on B-CLL cells. Transfection of B-CLL cells with these siRNA significantly diminished, or completely abrogated the expression of FIM mRNA as detected by RT-PCR. The figure below shows silencing of the FIM gene following siRNA transfection, as assayed by RT-PCR. “Untrans” and “ctrl” represents untransfected and control siRNA-transfected B-CLL cells respectively. As noted, transfection with each of the three siRNA against FIM completely abrogated the expression of FIM siRNA. 24–48 hours after siRNA transfection, a substantial fraction (30–70%) of the B-CLL cells, compared to cells transfected with control non-silencing siRNA, went into apoptosis as assayed by Annexin-V-propidium iodide staining. Time kinetic studies revealed that siRNA mediated silencing and apoptosis occurred between 18 and 48 hours. No such effect was noted when peripheral blood mononuclear cells of healthy donors or FIM-positive fibroblast cell lines were transfected with siRNA. In reconstitution experiments, siRNA treated B-CLL cells were co-cultured with various numbers of FIM-positive fibroblasts. The presence of these fibroblasts greatly diminished the apoptosis of B-CLL cells induced after siRNA treatment. Our results indicate that overexpression of FIM in B-CLL cells are critical for their survival and FIM may serve as a leukemia-specific target for B-CLL therapy.
Figure Figure
Efficacy wnd Bleeding with Ximelagatran and Warfarin after TKR: Pooled Subgroup Analysis of 3 Phase III Trials.
Authors: Berkowitz Scott D. , McElhattan Jennifer , Francis Charles W. , Colwell Clifford W. 
Published: October 2019

Abstract
Introduction: In patients undergoing total knee replacement (TKR), the post-operative period is a time of intense thrombotic stimulus often resulting in venous thromboembolism (VTE), precisely when surgical hemorrhage must be minimized: anticoagulant management requires a delicate balance. Ximelagatran (Exanta®, AstraZeneca), the first oral alternative to warfarin, has a rapid onset of action and requires no coagulation monitoring or dose adjustment. Three randomized, double-blind trials of ximelagatran vs. warfarin as VTE prophylaxis after TKR were completed: Study 236 (n=680), EXULT A (n=2301) and EXULT B (n=2303). Ximelagatran 36 mg BID provided efficacy superior to well-controlled warfarin (target INR 2.5) administered for 7–12 days, while 24 mg BID yielded numerically better but not statistically superior efficacy; neither dose increased bleeding.
Objective: We performed pooled subgroup analyses to assess pre-specified subgroup factors with regard to thromboembolic and adjudicated major/minor bleeding events.
Methods: Two comparison pools were created: 36-mg pool (ximelagatran 36 mg and corresponding warfarin groups from EXULT A and B) and 24-mg pool (ximelagatran 24 mg and corresponding warfarin groups from Study 236 and EXULT A). Prognostic factors were examined using multiple logistic regression analysis. Treatment by factor interactions are reported.
Results: In the 36-mg pool, no significant treatment interactions were observed for efficacy (i.e., the influence of each subgroup factor on ximelagatran and warfarin patients was similar). However, in the 24-mg pool, increases in age and CrCL indicated a disproportionate increase in the composite efficacy endpoint of total VTE and/or all-cause mortality for the 24-mg group compared to the warfarin group. Subgroup analyses of major/minor bleeding events showed no significant treatment interactions in the 24-mg pool. In the 36-mg pool, increases in age resulted in increased bleeding in the warfarin group compared to the 36-mg group. Men on ximelagatran 36 mg exhibited a greater risk of bleeding than did men on warfarin. No factors were identified to explain the observed increase in risk.

Conclusions: Subgroup analysis results for the ximelagatran 36-mg and 24-mg pools were generally consistent with those in the overall population, with the incidence of the composite efficacy endpoint consistently lower in the ximelagatran groups than in the warfarin group for most pre-specified subgroups. Patients expected to have higher ximelagatran exposure due to renal impairment and increased age showed no significant difference in bleeding risk compared with warfarin. This analysis supports a 36-mg dose for all patients undergoing TKR.
Monocyte-Derived Dendritic Cells from HLA-Matched Allogeneic Donors Showed Superiority To Induce Leukemic Cell-Specific T Cells in Comparison to Leukemic Cell-Derived Dendritic Cells or Monocyte-Derived Dendritic Cells from AML Patients.
Authors: Lee Je-Jung , Choi Bo-Hwa , Park Myong-Suk , Yang Deok-Hwan , Kim Yeo-Kyeoung , Kim Hyeoung-Joon 
Published: October 2019

Abstract
Leukemic-dendritic cells (leukemic-DCs) have certain limitations, which include difficult generation in 30–40% of patients, and low levels of expression of several key molecules. To overcome these limitations, autologous mononocyte-derived DCs (autologous-DCs) from the patients can use as an alternative approach, but the DCs may have some defective functions. In this study, we investigated the feasibility of immunotherapy for AML using leukemic-cell specific cytotoxic T lymphocytes that were stimulated by allogeneic monocyte-derived DCs (allogeneic-DCs) pulsed with leukemic cell lysates in vitro. To generate autologous- or allogenic-DCs, CD14+cells isolated from mononuclear cells (MNCs) of the patients or of HLA-matched donors were cultured in the medium with GM-CSF and IL-4 for 6 days. Their maturation was induced by adding TNF-α, IL-1β, IL-6, and PGE2 for 2 days, and then, leukemic cell lysates were pulsed. To generate leukemic-DCs, MNCs obtained from AML patients were cultured in the presence of GM-CSF, IL-4, and TNF-α for 8–12 days. The expressions of several key molecules on allogeneic-DCs were significantly higher than that on leukemic-DCs or autologous-DCs. Allogeneic-DCs exhibited a higher capacity to stimulate allogeneic CD3+ T cells compared to leukemic-DCs or autologous-DCs in allogeneic mixed lymphocyte reaction assay. Autologous CD3+ T cells stimulated by HLA-matched allogeneic-DCs pulsing with leukemic cell lysates showed more potent cytotoxic activities against autologous leukemic cells than those stimulated by leukemic-DCs or autologous-DCs. In addition, autologous-DC showed an intermediate level between leukemic-DCs and allogeneic-DCs in phenotypic expressions, allogeneic T-cell stimulatory capacities, or cytotoxic activities. In conclusion, these results suggest that monocyte derived-DCs from HLA-matched allogeneic donors can alternatively use to generate leukemic cell-specific cytotoxic T cells and to overcome the limitation of leukemic-DCs or autologous-DCs.
AML Induction Therapy with Outpatient Azacitidine.
Authors: Shadduck Richard K. , Rossetti James M. , Faroun Yacoub , Kaplan Robert B. , Lister John 
Published: October 2019

Abstract
15 patients with newly diagnosed acute myelogenous leukemia were treated with outpatient 5-aza-2′-deoxycytidine (azacitidine, 75 mg/m2 subcutaneously for seven days every four weeks) as primary induction therapy. The average age was 68 years (range = 44–80). Bone marrow blast counts ranged between 20% and 38%. The overall response rate (complete remission, CR + parital remission, PR) was 53% (8/15). CR was achieved in 4 patients (27%) in an average of 3 cycles (range = 2–5). Duration of response averaged 8 cycles (range = 6–13) from the time CR was documented. Two of these patients went on to allogeneic hematopoietic stem cell transplantation (one in CR, one in PR). PR was achieved in 4 patients (27%) in an average of 3 cycles (range = 2–3). Duration of response averaged 7 cycles (range = 1–15) from the time PR was documented. Two patients continue therapy, maintaining PR, at 9 and 15 cycles. One patient met criteria for stable disease with a survival of 17 months. One patient with biphenotypic leukemia survived 12 months with a significant reduction in transfusion requirements. One patient demonstrated a major platelet response, but discontinued therapy after 3 cycles due to anorexia. Four patients had disease progression after 1, 1, 2, and 3 cycles. The average survival for patients who achieved at least stable disease and did not go on to hematopoietic stem cell transplantation was 14+ months (range = 8–24). None of the responders had an ECOG performance status worse than 1 while being treated. The most common toxicity was febrile neutropenia (4 patients, 2 deaths). Azacitidine administered in the outpatient setting can induce remission in AML. This therapy appears to be well tolerated. Further study is warranted in poor risk patients to document activity and toxicity.
Organic Arsenic Lipid Derivatives Are More Potent and Less Toxic Than Inorganic Arsenic Trioxide in Preclinical Testing.
Authors: Cheng Xiaodong , Golemovic Mirna , Giles Francis , Zingaro Ralph , Gao Ming-Zhang , Freireich Emil J. , Andreeff Michael , Kantarjian Hagop M. , Verstovsek Srdan 
Published: October 2019

Abstract
Arsenic trioxide (ATO) is an inorganic arsenic derivative that is effective in patients with relapsed acute promyelocytic leukemia. It is being investigated as therapy for other cancers but the risk/benefit ratio for its use still has to be determined due to its significant side effects. In contrast, organic arsenic derivatives are known to be much less toxic. We have synthesized a series of eight organic arsenic lipid derivatives and have tested them in NIH 60 cell line screen. Based on its activity, we selected GMZ27 [3-S-di(n-propyl)arsino-3-thio-1,2-propanediol] for further study in our laboratory and have confirmed its potent activity against human acute myeloid leukemia cell lines HL60 and NB4, which was significantly higher than that of arsenic trioxide. The IC50 (concentration that kills 50% of the cells) in MTS assay is 0.5 and 0.9 μm for GMZ27. Assessment of the mechanisms of action of GMZ27 in cell lines has shown that the GMZ27 was more potent inducer of superoxide than ATO. GMZ27 caused dissipation of mitochondrial transmembrane potential, cleavage of caspase 9, caspase 3 activation, PARP cleavage, and compromise in cell membrane integrity. However, its treatment also resulted in caspase 8 cleavage, suggesting that it affects both intrinsic and extrinsic apoptotic pathway. Its activity was related to the level of glutathione in the leukemic cells as pretreatment of the cells with BSO, which depletes intracellular glutathione, results in cells sensitivity to GMZ27. On the other hand, pretreatment with DDT, which increases intracellular glutathione, results in their resistance to GMZ27. GMZ27 had no effect on cells maturation and differentiation, and cell cycle. GMZ27 when tested against healthy donor mononuclear cells in a colony forming assay showed significantly less toxicity than arsenic trioxide. In vivo toxicity testing in Swiss Webster mice showed LD50 (dose that kills 50% of mice) to be 100 mg/kg for GMZ27, comparing to 10 mg/kg for ATO. In conclusion, organic arsenic lipid derivatives, and in particular GMZ27, may have more potent antileukemic activity and significantly less toxicity in vivo and in vitro than ATO and therefore further development of these medications is warranted.
Monitoring Immune Responses Following Vaccination with Autologous Dendritic like Leukaemia Cells.
Authors: Klammer Matthias , Waterfall Martin , Samuel Kay , Thomas Charlotte , Johnson Peter R.E. , Turner Marc L. , Roddie Patrick H. 
Published: October 2019

Abstract
It has been demonstrated that in a proportion of patients with Acute Myeloid Leukaemia (AML) their leukaemic blasts can be induced to differentiate into Dendritic Like Leukaemic Cells (DLLC) in vitro under the influence of GMCSF, IL-4 and TNF alpha. DLLC hold promise as cellular vaccines aimed at eradicating Minimal Residual Disease following successful induction of remission by chemotherapy. In A Phase I/II Multi-Centre Study we assessed leukaemic blasts of 21 patients with AML for their cytokine driven potential to differentiate into DLLC in vitro. 7 out of 21 (33%) patients showed DLLC conversion of their AML blasts based on morphological appearances and immunophenotype. 5 patients have completed chemotherapy and were then eligible to receive 4 escalating doses of subcutaneous DLLC vaccine. Immune responses to the administration of the vaccine are monitored by a combination of methods. The emergence of leukaemia specific T-cells following vaccination is demonstrated using Elispot assaying of Interferon gamma release in an in vitro re-stimulation assay. In WT1 expressing patients HLA-Tetramers allow to determine the frequency of WT1 specific T-cells. Regulatory T-cells (CD4/CD25 positive) are monitored by flow-cytometry. Monitoring of Minimal Residual Disease (MRD) is undertaken by means of real-time quantitative RT-PCR for leukaemia specific fusion transcripts or WT1 gene expression.
4 patients have so far completed the DLLC vaccination course. Vaccination was well tolerated with minimal side effects. A more than two-fold increase of leukaemia specific cytotoxicity was demonstrated following DLLC vaccination in one patient, while reduction of MRD was seen in several patients during the vaccination. The increase in CD4 /CD25 positive regulatory T-cells observed in several patients post vaccination may serve to dampen the induced anti-leukaemic immunity.
A Phase II Trial of Busulfan, Melphalan, and Fludarabine followed by Allogeneic T-Cell Depleted Hematopoietic Stem Cell Transplants from HLA-Identical, or HLA-Non Identical Related or Unrelated Donors for the Treatment of Advanced Hematopoietic Malignancies.
Authors: Boulad Farid , Jakubowski Ann , Castro-Malaspina Hugo , Hsu Katharine C. , Kernan Nancy A. , Papadopoulos Esperanza B. , Perales Miguel A. , Prockop Susan E. , Small Trudy N. , van den Brink Marcel R.M. , Young James W. , Ro Michelle , Collins Nancy , O’Reilly Richard J. 
Published: October 2019

Abstract
Twenty five patients have been enrolled on this trial to date. There were 14 males and 11 females aged 0.6–54 years. Patients’ diagnoses and stage included: NHL in CR2 or refractory (n=2), AML (n=7), including 5 pts with secondary AML, ALL > CR3 (n=4), CML in CP2 (N=1) and high risk MDS (n=11) including 5 pts with secondary MDS. Eight pts had a matched related donor, 14 pts an unrelated donor and 3 pts a mismatched related donor. Cytoreduction consisted of busulfan (Bu) (0.8–1 mg/Kg/dose x 10 doses), melphalan (Mel) (70 mg/Kg/day x 2) and fludarabine (Flu) (25 mg/m2/day x 5). Graft rejection prophylaxis included rabbit ATG (Thymoglobulin) (2.5 mg/Kg/day x 2). Four pts tolerated only one of two doses of the ATG, 2 pts received equine ATG and one pt Alemtuzumab. Twenty one pts received G-CSF mobilized peripheral blood stem cell transplants that were T-cell depleted by CD34 selection and E-rosetting while the other four pts received Soybean agglutinin E-rosette depleted marrow grafts. Cell doses were 1.3–20.5 x 106 CD34 cells/Kg. and 0 -100 x 103 CD3 cells/Kg. Engraftment occurred in 24 pts. One pt suffered a graft failure; This pt had initial low busulfan levels, and received bone marrow derived stem cells with a low cell dose from a 5/6 HLA-matched unrelated donor. Acute graft-versus-host disease occurred in four pts: grade 1 (n=2) and grade 2 (n=2) and no pts developed any grade 3-4 severe GvHD. Two patients were diagnosed with chronic GvHD: localized (n=1) and extensive (n=1). Two patients developed sepsis early post BMT, with secondary multi organ failure and early mortality, while for the rest of the patients, regimen-related toxicity was acceptable. Relapse occurred in 9 pts. Mortality included 7 pts from relapse, two pts from sepsis and multi-organ failure, 3 pts from infections, and one pt from unknown causes. The overall survival (OS) and disease-free survival (DFS) at 2 yrs for the entire patient cohort were respectively 44% and 42 %; The DFS was 50% for patients with secondary MDS or AML. In summary, the cytoreduction with Bu Mel and Flu allowed consistent engraftment of T-cell depleted grafts and was associated with acceptable outcome for patients with secondary MDS or AML.
Reduced Intensity Conditioning Therapy Using Campath -1H Is Successful for Stem Cell Transplantation in Non-Malignant Disorders.
Authors: Rao Aarati , Hayashi Robert , Grossman William , Wilson David , Yu Lolie , DiPersio John , Barnes Yvonne , Shenoy Shalini 
Published: October 2019

Abstract
Stem cell transplantation (SCT), indicated for many non-malignant disorders, is limited by donor availability, graft rejection (GR), toxicities of conditioning, morbidity and mortality (TRM), and graft versus host disease (GVHD). To overcome these barriers, we tested a novel conditioning for SCT. It was designed to support engraftment by deleting host immune reactive lymphocytes and macrophages. Campath -1H (anti-CD52 mab) was given on days -21, -20, and -19 (total dose 48 mg), fludarabine (day -8 to -4) (total 150 mg/m2) and melphalan on day -3 (140 [n=15] or 70 [n=1] mg/m2). Stem cell sources were related/unrelated bone marrow (BM) (8), peripheral blood (PB) (5) and umbilical cord blood (UCB) (3). GVHD prophylaxis was cyclosporine (tapered after 3 months), methylprednisone (tapered after day +28) and methotrexate on days +1, +3, and +6. Methotrxate was not used in UCBT. End points of the study included engraftment and TRM. Sixteen patients (1.5–40 yrs) diagnosed with aplastic anemia (5), Hurler’s (2), sickle cell anemia, XLAAD, histiocytosis (3), thalassemia, adrenoleukodystrophy, Evan’s syndrome and dyserythropoietic anemia were transplanted. Median follow-up was 219 days (66–845). The regimen was well tolerated. All patients that survived >1 month engrafted. Neutrophils (ANC >500/dL) engrafted at a median of 12.5 (10–36) days and platelets (>50,000/dL) at a median of 21 (12 –63) days. Skin GVHD developed in 3 patients and resolved early. Four are tapering immune suppression; 8 are successfully off immune suppression. All survivors have either stable disease or are cured. One recipient had a normal pregnancy and delivered twins. Two patients died prior to engraftment from previously acquired Pseudomonas infection. A third patient died of CMV disease (day +112) and another recipient died of intracranial hemorrhage/refractory thrombocytopenia (day +43) after engraftment. Other complications were bacterial and viral infections occurring within the first 100 days post-transplant. All but 1 CMV+ recipient reactivated CMV as assessed by PCR. Profound lymphopenia was present in all recipients on day +30. NK cells recovered by day +100, CD8+ cells by day +180, CD4+ and B cells between days 180–270 after transplant. Immunoglobulin (Ig M and A) levels dropped post transplant and normalized by 9 months. Ig A recovered later than IgM. In summary, successful engraftment despite varied stem cell sources, was achieved without significant toxicity or GVHD. Lymphopenia resulted in a significant infection risk within the first 100 days post transplant, requiring close surveillance and early intervention. This transplant regimen is well tolerated and may preserve fertility, making it a promising alternative to conventional SCT.
Argatroban, Not Lepirudin or Bivalirudin, Treatment Results in the Generation of Nitric Oxide during Parenteral Administration.
Authors: Walenga Jeanine M. , Fareed Daniel , Schultz Chris , Neville Brian , Hoppensteadt Debra , Iqbal Omer , Fareed Jawed 
Published: October 2019

Abstract
While all direct thrombin inhibitors (DTIs) produce their anticoagulant effect by inhibiting thrombin, there are differences in their mode of action, not only in protease generation/inhibition, but also in modulating cellular mechanisms such as the release of endothelial mediators including nitric oxide (NO) and prostacyclin. Preliminary studies have revealed that argatroban treatment, unlike bivalirudin and lepirudin, results in vasomodulation and other pharmacological effects unrelated to thrombin inhibition [Pathophysiol Haemost Thromb2002;Suppl 3:56–65]. To validate this observation primates (Macaca mulatta; n=6/group) were treated with 100 to 500 μg/kg iv BID of these three DTIs. Blood samples were drawn at 5, 15, 30, 60, 120, 240 and 360 minutes. Total NO levels were measured by two different methods. The Griess reaction was performed by a chromogenic substrate method (R&D Systems, Minneapolis, MN) based on the two-step diazotization reaction where NO2 is acidified to produce a nitrosating agent, which is complexed with sulfanilic acid to produce a diazonium ion, which is then coupled with chromogenic reagents. A chemiluminescent assay was also used. Plasma NO reacts with oxygen to form nitrite. Using the Ionics Instrument 208I (Sievers Instrument, Boulder, CO), the nitrite is converted into NO by chemical reactions, and the resulting NO level is determined using a highly sensitive gas phase. DTIs did not have any non-specific interfering affect on the assays. Primates treated with argatroban showed a dose- and time-dependent increase in plasma NO levels. Baseline levels ranged from 4–8 μM (mean ± SD = 6.1 ± 1.8 μM), whereas 5 min after the administration of argatroban NO levels were 6–11 μM (7.2 ± 1.8 μM) for 100 μg/kg, 8–14 μM (11.8 ± 2.6 μM) for 250 μg/kg, and 11–19 μM (15.7 ± 3.1 μM) for 500 μg/kg argatroban. This increase in NO was observed for 30 min. The area under the curve (0–6 hrs) revealed a dose-dependent increase in a non-linear fashion. No difference between the two methods was noted. For lepirudin and bivalirudin, the NO levels did not differ from baseline during the study period, and no differences between these treatment groups were observed. Flow cytometric studies showed comparable inhibition (29–37%) of tissue factor mediated platelet activation 5 min after dosing for all DTIs. However, the % P-selectin expression was lower in argatroban treated primates than with lepirudin and bivalirudin treatment (17.1 ± 5.2 vs 29.3 ± 7.1; p <0.026). The 5 and 60 min argatroban samples assayed for 6-keto-PGF1 alpha showed 20–30% higher levels (not significant) compared to lepirudin and bivalirudin. These results support the hypothesis that argatroban is distinct from lepirudin and bivalirudin in that it produces an increase in NO and prostacyclin which may partly contribute to its therapeutic efficacy by stabilizing platelets.