bluuebunny/crossref_metadata_2025_split huggingface dataset:
Liver Fibrosis Assessment by Ultrasound Elastometry in Thalassemic Patients on Long-Term Iron Chelation.
Authors: Longo Filomena , Marletto Fabio , Rivetti Elisa , Sacchetti Laura , Piga Antonio 
Published: October 2019

Abstract
BACKGROUND
In thalassemic major patients liver fibrosis may be influenced by independent factors as hepatitis, degree of iron loading and iron chelation, and the specific effect of drugs. Due to its invasive nature, the histological evaluation of liver fibrosis on biopsy samples is difficult to be done and repeated. We analysed the relationship of these factors with the degree of liver fibrosis assessed non-invasively by ultrasound elastometry expressed as liver stiffness. We also compared the liver stiffness in patients on long term therapy with different iron chelators.
METHODS
We applied the ultrasound elastometry (Fibroscan®, Fibrosens, Paris) to a consecutive series of patients with ß thalassemia major on regular transfusion and chelation. Liver Iron Concentration (LIC) has been assessed by SQUID magnetic susceptometry (Tristan Technologies, Inc., San Diego, USA). We considered, as control, a group of healthy persons. All the subjects signed a written informed consent.
RESULTS
We studied 115 patients (64 males, 51 females) with beta thalassemia major, with mean age of 27.7 ± 7.6 years (range: 6.7–48.3). Eighty-six (75%) patients were HCV-Ab positive, and 52 (45%) HCV-RNA positive. Mean stiffness was 9.0 ± 6.6 KPa (range 3.3–43.5), significantly higher than in controls (4.0 ± 1.0, CI 3.3–4.8). Seven (6%) patients had a stiffness value above the cirrhosis threshold, and 18 (16%) were above the severe fibrosis threshold. Mean serum ferritin was 2064 ± 2011 μg/L, and mean LIC was 1845 ± 1199 microg/g ww. Considering the chelator, 34 patients have been on deferoxamine (DFO) therapy for 19.2 ± 9 years, 53 patients on deferiprone (L1) for 5.7 ± 3.7 years, and 28 on deferasirox (ICL670) for 3.3 ± 1.1 years. Mean stiffness was 9.5 ± 7.4 kPa, 9.9 ± 7.3 and 6.6 ± 2.8, respectively. On multivariate analysis, HCV Ab, HCV RNA and LIC were independent factors determining the degree of stiffness, while the chelator used was not significant.
CONCLUSIONS
Thalassemia major patients on regular transfusion and chelation have significantly abnormal degree of liver fibrosis. HCV positivity and the amount of iron overload seem to be the main related factors. The type of chelator used does not seem important.
Detachment of Human Endothelial Cells from Fibrinogen Induced by Cleaved High Molecular Weight Kininogen (HKa) Is Mediated by c-Jun Amino-Terminal Kinase (JNK) Signaling Pathway.
Authors: Liu Yuchuan , Sainz Irma M. , Pixley Robin A. , Colman Robert W. 
Published: October 2019

Abstract
We have shown that HKa inhibits angiogenesis by inducing detachment and apoptosis of human umbilical vein endothelial cells (HUVECs) from specific extracellular matrix proteins, such as vitronectin and fibrinogen, but not collagen. To investigate which intracellular signaling pathway led to cell detachment, several specific inhibitors were selected and utilized. SP600125, a JNK inhibitor, and PP2, a Src kinase inhibitor, both produced HUVECs detachment from surfaces coated with fibrinogen while MG-132, a NF-κB inhibitor, induced apoptosis. Neither U0126, an ERK inhibitor, nor SB202190, a p38 inhibitor, detached HUVECs from fibrinogen surfaces. By Western blot analysis, we found that HKa inhibited JNK, but not Src kinase activity. Previous investigators have demonstrated Rho-associated protein kinase (ROCK) regulates the JNK pathway. We demonstrated that Y27632, a ROCK inhibitor, not only inhibited JNK activity, but also caused detachment of HUVECs. Using phospho-antibodies, we found that HKa decreased the phosphorylation of paxillin at Ser178 site, but not Tyr 118 site indicating a serine kinase. Paxillin is a multi-domain adaptor found at the interface between the plasma membrane and the actin cytoskeleton. Paxillin phosphorylation is known to be associated with the coordinate formation of focal adhesions and stress fibres. Since the Ser178 of paxillin is a substrate for JNK, while Tyr 118 is a substrate for FAK and Src, it appears that JNK is responsible. Our studies have uncovered the novel finding that HKa detached HUVECs from fibrinogen through the ROCK-JNK-paxillin (Ser178) pathway, but not Src-paxillin (Tyr118) signaling.
Differential Gene Expression Patterns and Interaction Networks in BCR/ABL Positive and Negative Adult Acute Lymphoblastic Leukemias.
Authors: Juric Dejan , Lacayo Norman J. , Ramsey Meghan C. , Racevskis Janis , Wiernik Peter H. , Rowe Jacob M. , Goldstone Anthony H. , O’Dwyer Peter J. , Paietta Elisabeth , Sikic Branimir I. 
Published: October 2019

Abstract
BCR/ABL is associated with an unfavorable prognosis in adults with acute lymphoblastic leukemia (ALL). We used DNA microarrays to identify gene expression profiles and molecular interaction networks related to BCR/ABL status and clinical outcome in a set of 54 adult ALL specimens from the MRC UKALL XII/ECOG E2993 intergroup study (21 p185BCR/ABL- and 16 p210BCR/ABL-positive and 17 BCR/ABL negative). In order to avoid biases associated with commonly used sample amplification procedures, we have implemented an indirect two-step labeling protocol based on signal amplification by use of dendrimer technology. Using a two-class, non-parametric t-test and a false discovery rate cutoff of 5%, we identified 271 genes (including GAB1, CIITA, XBP1, CD83, SERPINB9, PTP4A3, NOV, LOX, CTNND1, BAALC and RAB21) as differentially expressed in BCR/ABL positive ALL compared with BCR/ABL negative ALL. They separate these two classes of adult ALL with an overall accuracy of 93% and are enriched for three highly relevant biological functions: Cellular Growth and Proliferation (57 genes, p = 0.004–0.044), Cell Death (49 genes, p = 0.0007–0.049), and Hematological System Development and Function (40 genes, p = 0.00004–0.049). Network analysis demonstrated complex interaction patterns of these genes, and identified FYN and IL15 as the hubs of the top-scoring network. We confirmed these findings by both qRT-PCR analysis of the initial set of samples and by cross-platform validation in an independent cohort of 128 adult ALL specimens. In addition, within the BCR/ABL positive subgroup, we identified 14 clones found to be over-expressed (TSPAN16, ADAMTSL4) or under-expressed (PILRB, STS-1, SPRY1) in p185BCR-ABL- relative to p210BCR-ABL-ALL. In a nearest-centroid classification, these clones correctly predict the BCR/ABL subtype with a cross-validated prediction accuracy of 95%. No differential gene expression was detected among Rho family GTPases and their known interaction partners. Finally, we constructed a gene expression- and interaction-based outcome predictor consisting of 27 genes (including GRB2, GAB1, GLI1, IRS1, RUNX2 and SPP1), which strongly correlated with overall survival in BCR/ABL positive adult ALL (p=0.0001), independently of other clinical parameters such as age (p=0.25) and white blood cell count at presentation (p = 0.003). These findings may be useful for developing novel therapeutic targets and prognostic markers in adult ALL.
Antibody Dependent Cellular Cytotoxicity (ADCC) Augmentation of Immunotoxin Activity in SCID-HSB-2 Mice Is Adoptively Transferable to NOD/SCID-HSB-2 Mice.
Authors: Flavell David J. , Flavell Sopsamorn U. 
Published: October 2019

Abstract
The anti-CD7 immunotoxin HB2-SAPORIN has significant therapeutic activity in the SCID-HSB-2 model of human T-ALL (Morland et al 1994 Br J Cancer 69, 279). In contrast the same immunotoxin shows virtually no therapeutic activity against the same tumour cell line in the NOD/SCID-HSB-2 model (Flavell et al 2000 Br J Cancer 83,1755). One difference between the two animal models is the presence of defective NK cells in NOD/SCID mice compared with the possession of fully functional NK cells by SCID mice. As we have previously shown that NK cell-mediated ADCC contributes significantly to HB2-SAPORIN activity in the SCID-HSB-2 model (Flavell et al 1998 Cancer Res 58, 5787) we hypothesise that it is defective NK cell function and a consequent reduction in ADCC in the SCID/NOD-HSB-2 model that explains the reduced therapeutic efficacy of immunotoxin in these animals. To test this hypothesis we adoptively transferred splenocytes from untreated or poly inosinic:cytidilic (poly I:C) treated SCID mice to NOD/SCID-HSB-2 mice prior to their treatment with immunotoxin. Adoptive transfer of un-stimulated splenocytes to NOD/SCID-HSB-2 mice did appear to improve the therapeutic performance of immunotoxin in these animals though this was not significant when compared to that observed in sham adoptively transferred animals. However, adoptive transfer of poly I:C activated splenocytes from SCID mice to NOD/SCID-HSB-2 mice did significantly improve the therapeutic performance of HB2-SAPORIN in the NOD/SCID-HSB-2 model. These preliminary data provide some further evidence for our hypothesis that it is deficient ADCC in the NOD/SCID-HSB-2 model that is responsible for reduced immunotoxin activity.
Effective Induction of Apoptosis by Mistletoe Plant Extracts in an Acute Lymphoblastic Leukemia Model.
Authors: Seifert Georg , Jesse Patrick , Prokop Aram , Reindl Tobias , Lobitz Stephan , Laengler Alfred , Lode Holger N. , Henze Guenter 
Published: October 2019

Abstract
Mistletoe (Viscum album) is one of the most used alternative cancer therapies applied as monotherapy or in combination with conventional therapies. Anti-tumor effects of mistletoe (MT) extracts were related to cytostatic and immunomodulatory effects observed in vitro. Aqueous MT extracts contain the three mistletoe lectins I, II and III as one predominant group of biologically active agents. The MT lectins inhibit protein biosynthesis by inactivating the 60S ribosomal subunit. Mistletoe lectin-I (ML-I) is one important apoptosis inducing compound. It is a heterodimer that consists of a cytotoxic A-chain (ribosome inactivating protein, RIP type 1) linked by a carbohydrate binding B-chain for cellular lectin uptake. However, although MT is widely used, there is a lack of scientific preclinical and clinical data. Here, we describe for the first time efficacy and mechanism of MT extracts against lymphoblastic leukemia in vitro and in vivo.
For this purpose, we first investigated both the cytotoxic effect and mechanism of action of two standardized aqueous MT extracts (MT obtained from fir trees (MT-A); MT obtained from pine trees (MT-P)) and isolated ML-I, in three human acute lymphoblastic leukemia (ALL) cell lines (NALM-6, sup-B-15 and REH). MT-A, MT-P and ML-I clearly inhibited cell proliferation as determined by LDH reslease assays at very low concentrations (ML-I LD50 from 0,05 ng/ml to 10 ng/ml depending on the host tree) with MT-P being the most cytotoxic extract. The mechanism of cell death was determined by DNA-fragmentation assays. These indicated dose dependent induction of apoptosis as the main mechanism of cell death.
Finally, we evaluated the efficacy of MT-A and MT-P in an in vivo SCID-model of pre-B ALL (NALM-6). For this purpose, mice (n=8/group) were injected i.v. with 1 × 106NALM6 cells and treated by intraperitoneal injections four times per week for 3 weeks (day 1–4; 7–11; 14–18) at varying doses (1, 5 and 50 mg/Kg (plant weight/body weight)). Mice (n=8) treated with PBS and cyclophosphamide (100 mg/kg, once on day 1) were used as negative and positive controls, respectively. Toxicity, peripheral blood counts, bodyweight and survival was determined over time.
Interestingly, both MT extracts in all tested concentrations significantly improved survival (up to 55,4 days) in contrast to controls (34,6 days). Furthermore, no hematologic side effects were observed from this treatment as indicated by completely stable blood counts. Also the body weight of treated animals remained stable over time indicating a complete absence of systemic toxicity in the selected dose range.
In summary, we demonstrate for the first time efficacy and mechanism of MT extracts against ALL in vitro and in vivo and hereby provide an important base line for the design of clinical trials with these compounds.
Outcome of Adult ALL Patients with Fusion Transcript E2A-PBX1: Results from the GMALL Therapy Trials 5/93 and 6/99.
Authors: Burmeister Thomas , Goekbuget Nicola , Schwartz Stefan , Hoelzer Dieter , Thiel Eckhard 
Published: October 2019

Abstract
The chromosomal translocation t(1;19) with fusion transcript E2A-PBX1 is the third most prevalent recurrent translocation in adult ALL (after t(9;22) and t(4;11)), accounting for about 3% of all cases. It’s prognostic impact in adults has been controversially discussed and it has been suggested that this aberration might define a high risk patient group. However, this would be in contrast to the situation in pediatric ALL, where E2A-PBX1/t(1;19) meanwhile defines a patient group with a better than average outcome. We have investigated retrospectively by RT-PCR 337 B-precursor BCR-ABL-/MLL-AF4-negative ALL patients enrolled in the GMALL 5/93 and 6/99 therapy trials for the presence of this mRNA fusion transcript. This represents the largest cohort of adult BCR-ABL-negative patients investigated for E2A-PBX1 by RT-PCR thus far. We assessed the clinical characteristics of the E2A-PBX1-positive patients and their overall survival.
Results: Eighteen patients were E2A-PBX1-positive by RT-PCR (5.8% of the investigated group, roughly 3% of the total of all ALL patients). Fourteen had a pre B (cyIg+), and 4 a common immunophenotype. Twenty-two percent of all pre B ALL patients were E2A-PBX1-positive. The median age of the positive patients at diagnosis was 27 (15–60) years and the median leukocyte count was 36.700/μl (N=17) with 7 of 17 patients presenting with an initial leukocyte count >30.000/nl (59% vs 27% in the negative group, p=0.005). Compared to the E2A-PBX1-negative group (239 common, 49 pre B, 31 pro B, median age 32 years, median leukocyte count 11.900 (N=312) the E2A-PBX1-positive patients had a lower overall survival rate (0.33 vs. 0.36). The CR rate did not differ significantly between E2A-PBX1-positive and -negative patients.
These results support the assumption that in contrast to the situation in pediatric ALL E2A-PBX1 defines a relatively unfavourable genetic subgroup in adult ALL. It is strongly correlated with high leukocyte count (>30.000/μl) at diagnosis which is a highly significant poor prognostic factor in B-precursor ALL. It remains to be established whether E2A-PBX1 can be confirmed as an independent prognostic factor within standard risk B-precursor ALL. E2A-PBX1 may be useful to separate in so called standard risk ALL a patient group which may benefit from stem cell transplantation in first CR.
Factor V Binding to Multimerin 1: Modulation by Factor V Activation and Binding Sites in the Factor V C1 and C2 Domains.
Authors: Jeimy Samira B. , Quinn-Allen Mary Ann , Fuller Nola , Segers Kenneth , Stafford Alan R. , Weitz Jeffrey I. , Camire Rodney M. , Nicolaes Gerry A.F. , Kane William H. , Hayward Catherine P.M. 
Published: October 2019

Abstract
Platelets and endothelial cells store the polymeric factor V(a) binding protein, multimerin 1 (MMRN1), for release upon agonist stimulation. In human megakaryocytes, factor V binding to MMRN1 follows plasma factor V endocytosis, resulting in stored complexes of MMRN1 and factor V in platelet α-granules. The C2 domain of the factor V light chain contains a MMRN1 binding site; however, the affinity and stoichiometry of factor V-MMRN1 binding have not been determined, direct comparisons of factor V and Va binding to MMRN1 have not been done, and potential homologous roles of C1 and C2 domain structures in MMRN1 binding have not been studied. To further explore the mechanism of factor V and Va binding to MMRN1, and the roles of B domain release and C1 domain residues in MMRN1 binding, we used surface plasmon resonance and solid-phase binding studies. Functional consequences of factor V-MMRN1 binding were tested in competitive binding assays with the soluble phospholipid 1,2-Dicaproyl-sn-glycero-3-phospho-L-serine (C6PS), and calibrated automated thrombinography (CAT). Factor V bound to MMRN1 with a higher affinity than factor Va (approximately 2 nM versus 12 nM), and a stoichiometry consistent with binding to MMRN1 trimers. The higher affinity of factor V for MMRN1 was mainly due to differences in rates of formation of a more stable, secondary complex with MMRN1. Factor V activation by thrombin dissociated bound factor V from MMRN1, consistent with the reduced affinity of factor Va for MMRN1. A panel of point mutated, B domain deleted factor V constructs were used to identify MMRN1 binding residues in the C1 domain of factor V and Va. On a three dimensional model of factor Va, these residues mapped to a large, predominantly contiguous region between the C1 and C2 domains, that overlapped residues critical for factor Va phospholipid binding and procoagulant function. Consistent with the lowered affinity of factor Va for MMRN1, C6PS significantly inhibited factor Va-MMRN1, but not factor V-MMRN1 binding (p<0.05). Overlap between the MMRN1 and phospholipid binding sites was verified by CAT assays, as MMRN1 caused dose-dependent, significant reductions in plasma thrombin generation in these assays, by increasing lag time (p<0.01), and reducing peak (p<0.01) and total thrombin generation (p<0.01). Taken together, these data indicate that the functional homologies between the C domains of factor V extend to their MMRN1 binding sites. Moreover, thrombin has modulating effects on factor V-MMRN1 binding that mimic its effects on factor VIII-von Willebrand factor binding. The affinity of factor V-MMRN1 binding could be important to promote the association of MMRN1 with factor V in platelets, until factor V release and activation for prothrombinase assembly.
CD15 (Lewis x) Expression in Human Myeloid Cell Differentiation Is Regulated by Sialidase Activity.
Authors: Gadhoum Samah Z. , Sackstein Robert 
Published: October 2019

Abstract
The cell surface carbohydrate Lewis x (Lex, CD15) is now well established as an important marker for immunophenotyping leukocytes and for immunoregulatory functions. Lex is related to sialyl Lex (sLex) by the addition of a sialic acid in the core Lex trisaccharide. This sialylation has profound implications as it confers novel biological function(s) to the core trisaccharide. In particular, expression of sLex correlates with E-selectin ligand activity. Though the expression of Lex in myeloid differentiation is now widely recognized, no studies to date have addressed the molecular mechanism regulating its expression. To examine this issue, we employed a leukemic model of differentiation based on the capacity of anti-CD44 mAbs to induce maturation and Lex expression. We found that anti-CD44 mAb-induced expression of Lex (> 30% increase in Mean Fluorescence Intensity (MFI) by flow cytometry) was accompanied by decreased expression (> 20% decrease in MFI) of its related structure sialyl Lex (sLex). Two non-mutually exclusive processes could account for the increase in Lex expression during myeloid differentiation:neo-synthesis of the Lex epitope and/orde-sialylation of the existing sLex.
To test whether increased expression of Lex during CD44-induced differentiation resulted from augmented synthesis, Acute Myeloid Leukemia (AML) cells were treated with anti-CD44 mAb in the presence of 4F-GlcNAc (2-acetamido-1,3,6-tri-O-acetyl-4-fluoro-D-glucopyranose), an inhibitor of poly-N-acetyl-lactosamine synthesis. 4-F-GlcNAc did not significantly alter the increase of Lex induced by the anti-CD44 mAb indicating that the CD44-mediated increase of Lex does not require de-novo polylactosamine synthesis. In presence of the sialidase inhibitor DANA (2,3-dehydro-2-deoxy-N-Acetyl-neuraminic acid) the anti-CD44-induced increase of Lex and decrease of sLex were abrogated indicating that sialidase plays an important role in the modulation of Lex expression during CD44 mAb-induced granulocytic differentiation. Moreover, using the exogenous substrate 4MU-NANA to measure endogenous sialidase activity in AML cells we found that the anti-CD44 mAb Hermes-1 induced a significant increase in the sialidase activity of AML cells, strongly suggesting that CD44-induced differentiation causes the elaboration of endogenous sialidases that are responsible for the modulation of Lex level by converting sLex into Lex. Furthermore, treatment of immature myeloid cells with G-CSF induced a 2-fold increase in sialidase activity, coincident with increased morphological changes of differentiation suggesting that the induction of surface sialidase activity is associated dynamically with the conversion of immature to mature phenotypes among native myeloid cells. Our results unveil a new paradigm, demonstrating a critical role for glycosidase activity in the “biosynthesis” of a key glycan determinant expressed on human cells.
p38 MAPK Signaling Mediates Retinoic Acid-Induced Expression of a Novel Tumor Suppressor Protein Programmed Cell Death 4 (PDCD4) in Acute Promyelocytic Leukemia Cells.
Authors: Akar Ugur , Ozpolat Bulent , Colburn Nancy , Lopez-Berestein Gabriel 
Published: October 2019

Abstract
Programmed-cell-death-4 (PDCD4) is a novel tumor suppressor protein that suppresses tumor promoter-induced neoplastic transformation. PDCD4 specifically inhibits the helicase activity of eukaryotic translation initiation factor 4A (eIF4A) and translation initiation and cap-dependent mRNA translation in vitro and in vivo. Loss or underexpression of PDCD4 is associated with carcinogenesis and chemoresistance in solid tumors. The role and regulation of PDCD4 in the the hematopoietic system and myeloid leukemia cells are not known. We previously reported that ATRA induces translational suppression through multiple posttranscriptional mechanisms during terminal cell differentiation (Harris et al, Blood, 104 (5) 2004). Therefore, in this study, we investigated the expression and regulation of PDCD4 during myeloid cell differentiation. All-trans-retinoic acid (ATRA) induces terminal differentiation in acute myeloid leukemia (AML) and promyelocytic (APL) cells, a well established model for myeloid cell differentiation. We found that treatment of HL60 (M2 type AML) and NB4 APL (M3 type AML) cells with ATRA (1 mM) induced PDCD4 protein and mRNA expression during granulocytic differentiation detected by western blot and RT-PCR analysis, respectively. We also demonstrated that inhibition of PDCD4 by siRNA reduced granulocytic differentiation induced by ATRA, suggesting that PDCD4 plays a role in granuliocytic differentiation. To determine mechanisms regulating PDCD4 we investigated the role of pP38 MAPK (Mitogen activated protein kinase) in reugulation of PDCD4 expression. ATRA induced PDCD4 expression correlated with activation of p38 MAPK (Mitogen Activated Protein Kinase) pathway in NB4 cells. To test the hypothesis that p38 MAPK signaling pathway mediates retinoic acid induced PDCD4 expression we treated cells with a specific p38 MAPK inhibitor, SB203580, ATRA or combination with ATRA. We observed that p38 inhibitor inhibited ATRA-induced expression of PDCD4 in NB4 cells. Basal level of PDCD4 expression was also markedly downregulated in the presence of p38 inhibitor when compared to untreated control cells, suggesting that p38 pathway is involved in ATRA-dependent and independent PDCD4 expression. Currently we are investigating whether inhibition of p38 by small interfering RNA (siRNA) will prevent expression of ATRA induced PDCD4 in APL cells. We are also trying to identify whether ATF2 transcription factor, a downstream of p38, is involved in PDCD4 expression. p38-mediated induction of PDCD4 pathway reveals a novel mechanism of PDCD4 regulation and ATRA action, providing a new insight into understanding terminal differentiation of myeloid cells. Better understanding the role of PDCD4 and posttranscriptional control of gene expression may offer targets for the differentiation therapy and chemo preventive strategies.