bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction of Localized TNF Activity Against Tumors Using Bispecific Antibodies.
Authors: Krusch Matthias , Herrmann Tanja , Kanz Lothar , Jung Gundram , Salih Helmut R. 
Published: October 2019

Abstract
Tumor necrosis factor (TNF) is a highly effective cytokine with considerable anti tumor activity not only because of direct tumoricidal effects but also due to its strong inflammatory and anti-angiogenic properties. However, while impressive clinical responses were obtained by locoregional application, e.g. in isolated limb perfusion, systemic application of TNF causes intolerable side effects. Therefore, multiple attempts have aimed for targeted delivery of TNF to maximize therapeutic efficiency and sparing normal tissue from detrimental effects, e.g. by generating antibody-derived fusion proteins with human TNF replacing the IgG1 CH2/CH3 Fc domain. However, a highly selective and thus clinically applicable drug for stimulation of TNF-receptors (TNF-R) at the tumor site with minimal unspecific TNF-R stimulation is still not available. Here we report the generation of a bispecific EpCAM X TNFR1 Fab2 antibody (bsFab2). The Fab fragments of an agonistic antibody to mouse TNFR1 were chemically hybridized to anti-EpCAM Fab fragment which served as a targeting molecule on EpCAM transfected mouse melanoma cells. Bound to such cells the bsFab2 exerted strong TNF activity as measured by killing of TNF sensitive WEHI-164 cells at concentrations as low as 0.1ng/ml. In the presence of untransfected tumor cells weak killing was detectable only at antibody concentrations as high as 1000ng/ml. Even more, killing of WEHI-164 cells in the absence of tumor target cells was undetectable. This demonstrates that TNF activity is induced by the bsFab2 in a target cell restricted manner. Target cell restriction was also confirmed when caspase activation rather than cell death was measured. We also demonstrate that the bsFab2 is able to induce target antigen restricted cellular inflammatory responses like monocyte/macrophage cytokine production. Next we investigated the activity of the bsFab2 in the B16 melanoma mouse model with syngeneic, immunocompetent mice. C57Bl/6 mice were inoculated 2500 EpCAM transfected B16 melanoma cells and treated with 6x20μg of the bsFab2. While all of six treated mice survived without developing a tumor, in the control group five of six mice died within 10 weeks. No significant side effects were observed. Our data demonstrate that the TNF activity of the EpCAM X TNFR1-bsFab2 is strictly depending on the presence of EpCAM positive tumor cells, stimulates cellular inflammatory responses in immediate vicinity of target antigen expressing tumors and provide the rationale for further studies evaluating the clinical potential of selective TNFR activation with bispecific antibodies in humans.
IVIG Down-Regulates LPS-Induced TNFα Production Via Inhibition of IκBα Activation: A Possible Mechanism of Action in the Treatment of Bacterial Infections.
Authors: Su Brenda B. , Chan Howard H.W. , Branch Donald R. , Denomme Gregory A. 
Published: October 2019

Abstract
Intravenous IgG (IVIG) preparations, originally developed as a substitution therapy for patients with low plasma IgG, are used in the treatment of various immune and nonimmune disorders including hemolytic uremic syndrome, necrotizing fasciitis, and idiopathic thrombocytopenic purpura (ITP); an autoimmune pro-inflammatory condition. However, the pleotropic effects of IVIG on Fcγ receptor-bearing immune cells are not fully understood. Bacterial lipopolyssacharide (LPS) is a principal pro-inflammatory signaling molecule recognized by the innate immune system. LPS binds to monocyte/macrophage Toll-like receptor 4 (TLR4) to activate transcription factors such as NF-κB/IκBα. In turn, NF-κB activation results in cytokine secretion such as tumor necrosis factor alpha (TNFα); a key pro-inflammatory cytokine that leads to monocyte apoptosis. Since IVIG reduces pro-inflammatory responses, we hypothesized that IVIG may down-regulate the LPS-induced pro-inflammatory response in monocytes. Using the human monocyte-like cell line THP-1 that expresses FcγRs and TLR4, we examined LPS-induced intracellular signaling after in vitro treatment with IVIG. Enzyme-linked immunosorbent assay (ELISA) on culture supernatants showed that 30 minute pre-treatment of THP-1 cells with IVIG (5 mg/mL final) downregulated subsequent LPS (200 ng/mL)-induced secretion of TNFα. Western blotting showed that IVIG increased Akt phosphorylation and resulted in phosphorylation of S6 ribosomal protein and Erk1/2 signaling molecules. The activation of S6 ribosomal protein was blocked by the PI3K inhibitor LY294002. More importantly, IVIG treatment inhibited LPS-induced IκBα activation; the likely reason for the decrease in LPS-induced TNFα secretion. In conclusion, IVIG down-regulates LPS-induced signal transduction and proinflammatory cytokine secretion, an action that is likely responsible for its usefulness in the treatment of bacterial infections and in other diseases where down-regulation of NF-κB is desirable.
A Retrospective Study of Current Anticoagulant Use in Patients with Atrial Fibrillation: Results from a Managed Care Database.
Authors: Murray Miranda , Friedman Howard 
Published: October 2019

Abstract
The vitamin K antagonist warfarin is currently the mainstay of anticoagulant therapy for patients with atrial fibrillation (AF), in whom it has been shown to be effective in preventing stroke. However, it is underused because, despite the quality of clinical care, warfarin can be a difficult drug to manage well. When initiating therapy, patients may require several visits to a coagulation clinic to achieve an international normalized ratio (INR) within the safe and effective range (INR 2–3). Thereafter, frequent monitoring of patients receiving warfarin may still be required due to intrapatient variability. Despite the benefits, studies have shown that approximately 50% of patients who should receive warfarin are not prescribed it or do not receive it. This study was performed to investigate the use of warfarin in eligible patients with AF, for whom it was indicated. Using the US PharMetrics database from 51 managed care organizations, data entered between January 1997 and April 2004, from 5940 patients (>18 years of age with a diagnosis of AF [ICD 9] for whom warfarin was indicated), were analyzed. Patients were excluded if they were not in the database 30 days before and 120 days after their initial diagnosis of AF. Patients were divided into four distinct cohorts, based on treatment patterns: steady users (at least 3 months of warfarin use); discontinued users (used warfarin for only 3 months); non-users (warfarin use for <30 days); and intermittent users (used warfarin prior to first diagnosis of AF, or had less than two prescriptions). Analysis showed that only 31% (1854) of patients were steady users, whereas 59% (3521) of patients were non-users. Also, 4% (246) and 5% (319) of patients were discontinued users and intermittent users, respectively. In addition to a diagnosis of AF, many patients had significant risk factors for stroke: hypertension (48%); diabetes (23%); congestive heart failure (21%); age 75 or older (19%); history of stroke or transient ischemic attack (7%). However, the study showed that patients prescribed warfarin did not have more risk factors compared with long-term steady warfarin users and warfarin non-users (see table). Overall, this study shows that warfarin is underused, even in patients with significant risk factors for stroke, which highlights the need for new, simpler, oral anticoagulant therapy.
Risk factors Total population [n (%)] Steady warfarin (%) Discontinued (%) Non-warfarin (%) Intermittent (%) NB Cumulative numbers of risk factors were constructed by assigning one point for each of the following: congestive heart failure; hypertension; age >75; diabetes; with two points added for prior stroke or transient ischemic attack 0 2036 (32) 32 29 36 34 1 1774 (30) 30 27 30 30 2 1197 (20) 22 23 19 23 3 623 (10) 11 13 10 8 4 204 (3) 3 4 4 4 5/6 106 (2) 2 3 1 1
Molecular and Phenotypic Analysis of Philadelphia Chromosome-Positive Bilineage Leukemia: Possibility of a Lineage Switch from T-Lymphoid Leukemia Progenitor to Myeloid Cells Via PU.1 Expression.
Authors: Nishii Kazuhiro , Monma Fumihiko , Lorenzo Felipe , Katayama Naoyuki , Shiku Hiroshi 
Published: October 2019

Abstract
The occurrence of acute bilineage leukemia is thought to be the malignant transformation of a myeloid or lymphoid leukemic progenitor with the potential to differentiate into the other lineages; however, the mechanisms of this lineage switch are not well understood. Here, we show the extremely rare case of adult Philadelphia chromosome positive acute bilineage leukemia, characterized by T-cell acute lymphoblastic leukemia, CD7+CD5+CD14−, and acute myelomonocytic leukemia, CD7−CD5−CD14+. Chromosome analysis showed 46,XY,del(7)(p11.2),t(9;22)(q34;q11.2) in all metaphase and leukemic cells expressed a minor BCR/ABL chimeric gene. When the CD5+CD14− and CD5−CD14+ cells were sorted, a fusion gene of BCR/ABL and a same clonal rearranged band of a T-cell receptor (TCR) gene were detected in both populations. Nucleotide sequencing of the TCRg gene revealed the clonal rearrangement of the V8-JGT2 complex in both populations. Over-expression of PU.1, which plays a fundamental role in myelomonocyte development was found in the sorted CD34+CD7+ and CD5−CD14+, but not CD5+CD14− cells. These results suggest that leukemic progenitor cells in the T-lineage with del(7),t(9;22) chromosome have the potential to differentiate into myeloid lineage and enforced PU.1 expression may contribute in part of this phenomenon. Studies of bilineage leukemia will be important for the understanding of lineage commitment and switch in hematopoietic cells.
Chromosome 21 Encoded RUNX1 and ETS-2 Overexpression in Regenerating Hematopoiesis in Children with Down Syndrome - Implications for Leukemiogenesis?.
Authors: Reinhardt Dirk , Wortmann Kristin , Kolar Miriam , Klusmann Jan H. , Puhlmann Ulrike , Bourquin Jean P. , Creutzig Ursula , Langebrake Claudia 
Published: October 2019

Abstract
Children with Down Syndrome (DS) are at an 150 fold increased risk to develop acute megakaryoblastic leukemia (AMKL) within the first 4 years of life. About 10% of newborns with trisomy 21 showed transient myeloproliferative disorder (TMD). Although mutations of the transcriptional factor GATA1, resulting in the shortened GATA1s have been shown in almost all blasts in DS-AMKL and TMD the predisposition to leukemiogenesis related to trisomy 21 is not clear. TMD occurred during embryonic stress hematopoiesis leading to the hepatic proliferation of the GATA1s positive blasts. Typically blasts disappeared within the first 3 month of live, however after a median time of 1.3years (0.6 to 3.7 years) 20% of the children suffered AMKL and required intensive cytostatic treatment.
The expression of chromosome 21 encoded hematological transcription factors (TFs) such RUNX1, ETS-2 and ERG were analysed in leukemic blasts from DS- TMD(n=7), DS-AMKL (n=25), DS without hematological disorder (n=10), AMKL (n=10) and healthy controls (n=7) by qRT-PCR.
Results: No increase of RUNX1, ETS-2 and ERG expression could be shown. By contrast, ERG was decrease in all leukemias and in DS without hematological disorder (p Anova.<0.002). GATA1s was significantly overexpressed in TMD and DS-AMKL (pAnova <0.02), whereas GATA1 expression in AMKL and controls was not changed. GATA2 was elevated (pAnova <0.01) in all megakaryoblastic leukemias, with or without DS (pAnova <0.0001). PU.1, typically associated with early lymphatic differentiation and granulopoiesis was down regulated in all megakaryoblastic leukemias and, surprisingly, in DS without hematological disorder. This confirmed previously reported results by gene-array analysis1.
To get further insight in the predisposition caused by trisomy 21 we analysed regenerating hematopoiesis in DS (n=14) partly resembling embryonic stress hematopoiesis. Correlated to the amount of bone marrow activation (CD38 positivity) a myeloid cell population (CD13/CD33 positive); with the co-expression of CD56 (NCAM) and CD36 (thrombospondin-receptor) could be detected by immunophenotyping (median percentage all nucleated bone marrow cells: 73±10%). In children without DS but regenerating hematopoiesis (n=41) a similar population of 4.6±1.8% (p<0.00001) could be detected.
For further analysis the CD33/CD56 positive cells were sorted (FACSVantage). The cells showed normal myeloid morphology and differentiation, lack of GATA1s mutation, but an aberrant TF expression pattern. RUNX1 was 10-fold and ETS-2 5-fold higher expressed compared to controls (p<0.012).
Summarized, (1) DS-AMKL and TMD leukemic blasts showed no general gene-dosage effect. However, (2) in stimulated bone marrow (stress hematopoiesis) trisomy 21 led to an overexpression of chromosome 21 encoded TFs, which might contribute to leukemiogenesis.
Human BCL11B Is Expressed in Normal T Cells and Differentially Expressed in T-Cell Malignancies.
Authors: Pulford Karen , Lyne Linden , Roncador Giovanna , Kominami Ryo , Banham Alison H. 
Published: October 2019

Abstract
B-cell lymphomaleukaemia 11B (BCL11B), the human homologue of murine Bcl11b/Rit-1/CTIP2, was originally identified as a novel tumor suppressor gene in a study of gamma-radiation induced thymic lymphomas in mice. Later studies, however, showed Bcl11b to have a vital role in T cell development and survival, with BCL11B translocations involving a variety of partner genes being reported in both T-cell and myeloid leukemias. The human BCL11B gene, located at 14q32.2, encodes an 832aa Kruppel C2H2 zinc finger protein that is functionally uncharacterized but likely to act as a transcriptional regulator. Analysis of publically available normal tissue Affymetrix microarray expression data indicates an expression pattern restricted to hematopoietic cells, with high levels of BCL11B transcripts being present only in peripheral blood T cells, NK cells, thymus and tonsil.
We have used two rabbit polyclonal anti-BCL11B antibodies to study the distribution of BCL11B protein in both normal and neoplastic human cells. These reagents, raised against two distinct regions of the murine Bcl11b protein (zinc finger and C-terminus), recognized the human BCL11B protein. While neither antibody stained B-cells in tonsil, one was crossreactive with the highly homologous BCL11AXL protein by Western blotting. In normal tissues, BCL11B protein expression was confined to the nuclei of the vast majority of T cells in thymus (foetal and adult) and tonsil. High levels of BCL11B were detected in T-cell lines, including the Molt-4, CCRF-CEM and Jurkat T-cell acute lymphoblastic leukaemia (T-ALL) derived cell lines. No expression was detected in any B-cell derived (pre-B to plasma cell stage) or myeloid cell lines studied. These results are consistent with the microarray gene expression data. In T-cell malignancies, BCL11B protein was only detected in a proportion of tumors, including 5/6 T-ALLs (one being weakly stained) and 2/8 peripheral T-cell lymphomas (weak cytoplasmic staining only). Interestingly, no expression was detected in ALK-positive anaplastic large cell lymphoma lines or tumors. Further studies of a larger series of T-cell malignancies are in progress. All other tumors studied, including B-ALL, chronic lymphocytic leukaemia, diffuse large B-cell lymphoma, mantle cell lymphoma, Burkitt’s lymphoma, follicular lymphoma, myeloma and Hodgkin’s lymphoma, were unlabelled.
In conclusion, the distribution pattern of the BCL11B protein in a wide range of both normal and neoplastic tissues is described for the first time. The study of BCL11B expression is an invaluable first step towards elucidating the role of this protein in T-cell biology and the significance of its differential expression in T-cell malignancies.
G1 Cell-Cycle Arrest and Apoptosis by Histone Deacetylase Inhibition in MLL-AF9 Acute Myeloid Leukemia Cells Is MLL-AF9 Independent.
Authors: Tonelli Roberto , Sartini Roberta , Fronza Raffaele , Freccero Francesca , Franzoni Monica , Dongiovanni Danilo , Ballarini Marco , Ferrari Simona , D’Apolito Maria , Di Cola Gabriella , Capranico Giovanni , Khobta Andriy , Campanini Renato , Paolucci Paolo , Minucci Saverio , Pession Andrea 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) with MLL rearrangements (MLLmut), found mainly in M5 or M4 FAB subtypes, is frequent in infants and secondary leukemias. The most common MLL translocation gives rise to MLL-AF9. MLL protein interacts with histone deacetylases (HDACs) -1 and -2 through the MLL repression domain.
We report the effects of HDAC inhibition by valproic acid (VPA) in MLL-AF9 AML-M5 cells (THP-1, MM6 and MOLM-13) and MLLmut AML-M5 blasts. VPA led to histone hyper-acetylation, strong cell-growth inhibition, G1 cell-cycle arrest and apoptosis. Combined treatment with all-trans-retinoic-acid (ATRA) did not substantially improve these effects. VPA increased MLL-AF9 transcription, indicating that VPA overcomes the cell-growth promoting activity and resistance to apoptosis conferred by MLL-AF9 in AML-M5 cells, even with increased MLL-AF9. A small number of genes were significantly affected by VPA in p53-absent THP-1 cells (GeneChip analysis), and the majority of these were up-regulated. VPA potently induced p21 and cyclin G2 (CG2) expression. p21 and CG2 targeted inhibition demonstrated that p21 acts as a key regulator in the VPA-inducted G1 cell-cycle arrest, while induction of CG2 has no effect.
These data suggest that these poor prognosis patients may benefit from HDAC inhibitor therapy.
In Acute Leukemia, the Polymorphism −211C>T in the Promoter Region of the Multidrug Resistance-Associated Protein 3 (MRP3) Is Not Associated with the Expression Level of the Gene.
Authors: Dorfel Claus , Sauerbrey Axel , Gruhn Bernd , Zintl Felix , Steinbach Daniel 
Published: October 2019

Abstract
We could recently show that the expression of the multidrug resistance-associated protein 3 (MRP3; ABCC3) is an important prognostic factor in ALL and in AML (Blood2003; 102:4493–8 and Clin Cancer Res2003; 9:1083–6).
Lang et al. showed that the single nucleotide polymorphism −211C>T (NCBI SNP ID: rs4793665) in the promoter region of the MRP3 gene is associated with the expression of the gene in liver cells (Pharmacogenetics2004;14:155–64). They found that individuals who were homozygous for cytosine in position 211 showed higher levels of MRP3 mRNA and protein. Lang et al. also provided evidence that the polymorphism is located at a binding site for transcription factors and that the polymorphism affected the binding of such factors. This could explain the influence of the genotype on the transcription level of the gene.
Because of these findings, we hypothesized that the SNP −211C>T might also determine the expression of MRP3 in acute leukemia and therefore be an inborn determinant of drug resistance. In order to prove this hypothesis we analyzed this SNP in our previously described groups of ALL and AML patients.
The genotype was determined using a ready to use TaqMan® SNP Genotyping Assay and the ABI PrismTM 7700 Sequence Detector (Assay ID: C_27829307_10; Applied Biosystems, Foster City, CA, USA).
Samples from 139 patients were analyzed. The frequency of the genotypes was: TT = 32% (n=44), CC = 22% (n=31), and CT = 46% (n=64). We did not find any correlation between the genotype and the expression of MRP3 (p=0.44). The result was the same, when patients with AML, T-lineage ALL, and precursor B-lineage ALL were analyzed separately. We also did not find a correlation between the genotype and response to therapy or the chance of survival.
In conclusion, the SNP −211C>T in the promoter region of MRP3 does not have a major impact on the level of MRP3 gene expression in acute leukemia and it does not determine the response to therapy. Possibly, the putative transcription regulator which binds at this part of the MRP3 gene is not active in acute leukemia.
Arsenic Trioxide Induces Regulated, Death Receptor- and Caspase-Independent Cell Death through the Mitochondrial Death Pathway.
Authors: Scholz Christian , Richter Antje , Lehmann Mario , Schulze-Osthoff Klaus , Dörken Bernd , Daniel Peter T. 
Published: October 2019

Abstract
Arsenic trioxide (As2O3, arsenite) efficiently kills cells from various hematologic malignancies and has successfully been employed especially for the treatment of acute promyelocytic leukemia. There, and in lymphoid cells we demonstrated that arsenite induces cell death in a caspase-2 and -9-independent fashion. Here, we address a potential role of death receptor signaling through the FADD/caspase-8 death inducing signaling complex in arsenite-induced cell death. In detail, we demonstrate that arsenite induces cell death independently of caspase-8 or FADD and cannot be blocked by disruption of CD95/Fas receptor ligand interaction. Unlike in death receptor ligation-induced apoptosis, As2O3-induced cell death was not blocked by the broad spectrum caspase inhibitor z-VAD-fmk or the caspase-8-specific inhibitor z-IETD-fmk. Nevertheless, arsenite-induced cell death occurred in a regulated manner and was abrogated upon Bcl-2 overexpression. In contrast, arsenite-induced cell demise was neither blocked by the caspase-9 inhibitor z-LEHD-fmk nor substantially inhibited through the expression of a dominant negative caspase-9 mutant. Altogether our data demonstrate that As2O3-induced cell death occurs independently of the extrinsic death receptor pathway of apoptosis. Cell death proceeds entirely via an intrinsic, Bcl-2 controlled mitochondrial pathway that does, however, not rely on caspase-9.
Apoptotic Cross-Talk between Organelles Induced by Fenretinide in Hematopoietic Malignancy.
Authors: Wang Kankan , Xiao Dakai , Du Yanzhi , Fan Huiyong , Chen Yulong , Chen Saijuan , Chen Zhu , Zhang Ji 
Published: October 2019

Abstract
Fenretinide (N-4-hydroxyphenyl retinamide, 4-HPR), a synthetic analogue of all trans retinoic acid (ATRA), has shown promising potentials in chemopreventive/therapeutic treatment of human malignancies due to its minimal toxicity, ability to control tumor growth in animal models, and synergism with a number of other anti-cancer agents. Clinical trials of fenretinide, as a preventive agent, have proven its efficacy in a number of tumor types including breast cancer, ovary carcinoma, prostate cancer, neuroblastoma and leukemia. In contrast to ATRA which induces cell differentiaion, fenretinide induces cell apoptosis in a variety of tumor cell lines. Although intensive studies have revealed that fenretinide may trigger apoptosis via induction of different signal molecules such as reactive oxygen species (ROS) and ceramide, mechanisms underlying fenretinide induced apoptosis are far from clear, which is largely due to the fact that a multitude of responses occur in cells upon fenretinide treatment. Accordingly, we have applied an approach integrated with cDNA microarray, advanced technology of data and literature mining, and methods of protein biochemistry to study the process of fenretinide-induced apoptosis in leukemia cell line NB4 in detail. Numerous novel features with the temporospatial relationship were revealed, indicating the coordinated regulation of molecular network from various aspects of cellular activities to program cell death. These features include changes in transcriptional regulation, translational regulation, signal transduction, metabolism and stress response. In consequence, cells treated with fenretinide appear to enter programmed cell death through complex cross-talk between organelles such as mitochondria and endoplsmic reticulum. This study may therefore have substantial impact on the development of more sophisticated protocols in leukemia and probably other malignancies as well.