bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anticoagulant and Anti-Inflammatory Effects of Recombinant Human Antithrombin in Man after LPS Challenge.
Authors: Jilma Bernd , Leitner Judith M. , Mayr Florian B. , Firbas Christa , Reiter Rosemarie A. , Kalhs Peter 
Published: October 2019

Abstract
Background: Antithrombin (AT) had beneficial effects on mortality of septic patients in an a priori defined subpopulation of the Kypersept trial, which received no concomitant administration of heparin.
Objectives: We hypothesized that recombinant human (rh)AT (without concomitant heparin) has anticoagulant properties and may decrease cytokine production in a well standardized model of human endotoxemia.
Methods: This study was randomized, double-blind, placebo-controlled in parallel groups in 30 healthy male volunteers. The active treatment groups received bolus primed continuous infusion of rhAT (recombinant human Antithrombin) to increase AT-levels to 200% and 500% iv. before infusion of 2ng/kg endotoxin
Results: Infusion of rhAT rapidly decreased neutrophil (p<0.01) and monocyte counts (p<0.05) before LPS-challenge, demonstrating that rhAT directly interacts with these leukocyte subsets. rhAT exhibited intrinsic anticoagulant efficacy ex vivo as shown by thrombelastography ((p<0.01). More importantly, in vivo thrombin formation decreased in a dose dependent fashion as measured by prothrombin fragment (F1+2) and thrombin antithrombin complexes (TAT). rhAT significantly decreased interleukin-6 (IL-6) release by 40%.
Conclusion: rhAT exhibited intrinsic anticoagulant effects, which are stronger than the minor effects of drotrecogin alfa (rhAPC; Blood2003;102:2093ff) in this model of tissue factor triggered coagulation. Effects of rhAT on leukocytes and inhibition of IL-6 release were not previously observed with various different synthetic or natural anticoagulants in this model and therefore represent specific pharmacodynamic properties of rhAT. The demonstrated anticoagulant and anti-inflammatory properties of rhAT may have beneficial effects in critically ill patients, and may explain the efficacy of rhAT in septic patients without concomitant heparin administration.
P53 and p21waf1— Expression by Immunohistochemistry in Diffuse Large B-Cell Lymphoma Has a Strong and Independent Impact on Survival of Patients with Germinal Center Phenotype.
Authors: Visco Carlo , Canal Fabio , Andreoli Annalisa , Parolini Claudia , Lestani Maurizio , Ambrosetti Achille , Krampera Mauro , Pizzolo Giovanni , Chilosi Marco 
Published: October 2019

Abstract
Over-expression of P53-related protein detected by immunohistochemistry (IHC) and absence of p21waf1— expression, the main downstream target following P53 activation, are useful surrogate markers in identifying diffuse large B-cell lymphoma (DLBCL) patients with mutated P53-gene. Following recent studies on gene expression, tissue micro-array technology has been shown to represent a reliable technique in discriminating DLBCL with germinal center B-cell like (GC), or post- germinal center (non-GC) origin.
We analyzed 80 consecutive patients with de-novo DLBCL, homogeneously treated with cure-intent (CHOP-like regimens +/− Rituximab) in a single Institution, for expression of p53/p21waf1—, with regard to the expression of GC (CD10+, bcl-6+/mum-1−/CD138−), and post-germinal center (CD10−, mum-1+, CD138+) IHC markers. Cases with 50% or more p53-positive neoplastic cells were defined as p53+++. All patients were provided with complete clinical information. Median age of our patients was 50 years, AAS was III or IV in half of patients, IPI was high or int/high in 36%. Median follow-up for survivors was 77 months. A p53+++/p21waf1— phenotype (corresponding to loss of function of the P53-gene) was detected in 19 of 80 patients (23%), while a GC phenotype characterized 42 patients (52%). Patients with P53+++/p21waf1— phenotype were more frequently resistant to induction therapies (p<0.0001) and had a significantly lower 5-year Progression Free Survival (PFS, 27% vs 48%, p=0.01). Patients with GC phenotype were associated with a better 5-year PFS than non-GC patients (57% vs 26%, p=0.02). When analysis was restricted to the 42 patients with GC phenotype, p53+++/p21waf1— phenotype could discriminate 28% of patients with PFS of 26% vs 68% of patients with GC pattern and other p53/p21waf1— phenotypes (p=0.005). In contrast, the outcome of patients with a non-GC phenotype was not modified by p53/ p21waf1— expression. At multivariate analysis, p53+++/p21waf1— phenotype, non-GC phenotype, and high IPI resulted the strongest independent factors in worsening PFS of our patients. Combining such adverse prognostic factors we could identify 60% of patients with an extremely poor prognosis (5-year PFS of 28%).
Our results strongly point to the usefulness of a combined molecular and clinical approach for prognostic considerations in DLBCL, and should be validated in larger series.
ReSETting PP2A Tumor Suppressor Activity Overcomes BCR/ABL Leukemogenic Potential in Blast Crisis CML.
Authors: Neviani Paolo , Santhanam Ramasamy , Trotta Rossana , Notari Mario , Blaser Bradley W. , Chang Ji-Suk , Liu Shujun , Mao Hsiaoyin , Oaks Joshua J. , Roy Denis C. , Valtieri Mauro , Bruner-Klisovic Rebecca , Caligiuri Michael A. , Bloomfield Clara D. , Marcucci Guido , Perrotti Danilo 
Published: October 2019

Abstract
A tight control of kinase and phosphatase activity is fundamental for normal cell growth, survival and differentiation. The deregulated kinase activity of the BCR/ABL oncoprotein is responsible for the emergence and maintenance of chronic myelogenous leukemia (CML). By contrast, PP2A, a serine-threonine phosphatase involved in the regulation of many cellular functions, was found genetically inactivated in many types of cancer. We show here that, in BCR/ABL-transformed cells and CD34+ CML blast crisis progenitors, the phosphatase activity of the tumor suppressor PP2A is inhibited by the physiological PP2A-inhibitor SET whose expression is enhanced by BCR/ABL and increased in blast crisis CML. In imatinib-sensitive and -resistant (T315I included) BCR/ABL+ cell lines and in CD34+ CML blast crisis cells, molecular and/or pharmacological activation of PP2A leads to dephosphorylation of important regulators of proliferation and survival of CML progenitors, suppresses BCR/ABL kinase activity and promotes BCR/ABL proteasome degradation via a mechanism that requires the SHP-1 tyrosine phosphatase activity. Furthermore, PP2A activation achieved by shRNA-mediated SET knock-down or PP2Ac overexpression or treatment with the PP2A activator forskolin results in growth suppression, enhanced apoptosis, restored differentiation, impaired clonogenic potential and decreased in vivo leukemogenesis of wild type and T315I BCR/ABL-transformed myeloid cells. Thus, functional inactivation of PP2A phosphatase activity is essential for BCR/ABL leukemogenesis and, perhaps, required for transition of CML into blast crisis.
A New Abl Kinase Inhibitor (AMN107) Has In Vitro Activity on Chronic Myeloid Leukaemia (CML) Ph+ Cells Resistant to Imatinib.
Authors: Martinelli Giovanni , Martelli Alberto M. , Grafone Tiziana , Mantovani Irina , Cappellini Alessandra , Tazzari Pier Luigi , Ottaviani Emanuela , Soverini Simona , Amabile Marilina , Renzulli Matteo , Terragna Carolina , Colarossi Sabrina , Iacobucci Ilaria , Poerio Angela , Baccarani Michele 
Published: October 2019

Abstract
Imatinib mesylate (Novartis Pharma), an inhibitor of the bcr/abl tyrosine kinase, has rapidly become the first-line therapy for CML. Imatinib has proved remarkably effective at reducing the number of leukaemia cells in individual CML patients and promises to prolong life substantially in comparison with earlier treatments. However, in patients in advanced phases of the disease, the development of resistance to this drug is a frequent setback. Therefore, new inhibitors of bcr/abl are needed. Very recently, a new bcr/abl inhibitor, AMN107 (Novartis Pharma), has been developed. We have tested AMN107 on human leukaemia cell lines and on blasts isolated from imatinib-resistant CML patients. After a 24 h incubation, AMN107 (10 nM) blocked K562 cells in the G1 phase of the cell cycle. To obtain the same effect with imatinib, a 200 nM concentration was required. AMN107 had no affect on cell cycle progression of bcr/abl-negative cell lines such as HL60 and NB4, even if the concentration was raised to 500 nM. After 48 h incubation, AMN107 (10 nM) was capable of inducing a massive apoptosis of K562 cells whereas, once again, 200 nM imatinib was required to obtain the same effect.
Western blot analysis with phosphospecific antibodies revealed that in K562 cells AMN107 (50 nM) markedly down-regulated autophosphorylation of bcr/abl Tyr177 and Tyr412, whereas autophosphorylation of Thr735 was unaffected. In contrast, imatinib even if used at 200 nM, did not diminish phosphorylation of either bcr/abl Tyr177 or Tyr412. This finding seems particularly important because recent evidence has demonstrated that the signalling pathway emanating from Tyr177 plays a major role in the pathogenesis of CML. Indeed, phosphorylated Tyr177 forms a high-affinity binding site for the SH2 domain of the adapter Grb2. The main effectors of Grb2 are Sos and Ras, however Grb2 also recruits the scaffolding adapter protein Gab2 to bcr/abl via a Grb2-Gab2 complex, which results in activation of phosphoinositide 3-kinase (PI3K)/Akt and Erk signalling networks. Consistently, we found by immunoprecipitation decreased levels of bcr/abl-associated Gab2, Grab2, and p85 regulatory subunit of PI3K in AMN107-treated cells. AMN107 treatment of K562 cells also caused a reduction of STAT5, cCBL, CRKL, and Akt phosphorylation levels, as well as Bcl-XL expression. AMN107 (5 μM for 24h) significantly increased the apoptosis rate of CML blasts isolated from patients resistant to imatinib. Therefore, AMN107 might represent a new bcr/abl selective inhibitor useful for overcoming imatinib resistance.
FOXO Transcription Factors Are Negatively Regulated by p38 Map Kinases Downstream of FLT3 Receptor Signaling.
Authors: Scheijen Blanca , Ngo Hai T. , Griffin James D. 
Published: October 2019

Abstract
The phosphoinositide 3-kinase (PI3K)-AKT/PKB signal transduction cascade is an evolutionarily conserved pathway regulating the FOXO subclass of Forkhead transcription factors (FOXO1, FOXO3a and FOXO4) downstream of insulin, cytokine and growth factor receptors. When PI3K phospholipid kinase activity is low, FOXO proteins localize to the nucleus where they induce expression of genes that promote programmed cell death (FasL, Bim, TRAIL), inhibit cell cycle progression (p27KIP1, Rb related family member p130), facilitate DNA repair (GADD45) and protect against oxidative damage (MnSOD). Following receptor stimulation, increased levels of PtdIns(3,4,5)P3 are generated at the plasma membrane resulting in PDK1-mediated activation of the serine/threonine kinase AKT/PKB. Subsequent phosphorylation of FOXO proteins by AKT/PKB results in their active sequestration in the cytoplasm through binding to 14-3-3 proteins, thereby blocking their action as transcription factors. Recently, it has become evident that FOXO proteins can also be regulated through other signaling pathways. One example is phosphorylation and activation of FOXO4 by stress kinases JNK/SAPK in response to oxidative stress (Essers et al., 2004; EMBO J. 23:4802–12).
Here, we report that p38 mitogen activated protein kinases (MAPK) negatively regulate FOXO transcription factors downstream of FLT3 receptor tyrosine kinase signaling. Stimulation of human wild-type FLT3 receptor with FLT3 ligand, or expression of constitutively active mutants FLT3-ITD or FLT3-D835Y in Ba/F3 cells resulted in activation of p38 MAPK and phosphorylation of its known substrate ATF2 on threonine residue 71 (Thr71). Transient luciferase reporter assays showed that expression of a constitutively active MKK3 or MKK6, upstream activators of p38 MAPK, inhibited FOXO-induced gene transcription. The p38 MAPK inhibitors SB202190 and SB203580 as well as dominant-negative p38 MAPK blocked FLT3-mediated inhibition of FOXO transcription activation, suggesting p38 MAPK is both necessary and sufficient for FOXO regulation. In vitro kinase assays demonstrated that FOXO proteins are directly phosphorylated by p38 MAPK, although different p38 MAPK family members (p38α, p38β 2, p38γ and p38δ) displayed distinct FOXO protein substrate specificity. In conclusion, our data demonstrate that FOXO proteins are novel substrates of p38 MAP kinases and activation of p38 MAPK through FLT3 receptor signaling represents an important pathway to attenuate FOXO target gene regulation.
Directed Donor Cord Blood Transplantation for Hematological Disorders.
Authors: Walters Mark C. , Quirolo Lynn , Edwards Sandie , Lee Joanna , Robertson Shanda , Saba Julie D. , Trachtenberg Elizabeth , Quirolo Keith C. , Lubin Bert 
Published: October 2019

Abstract
The Sibling Donor Cord Blood Program was initiated in 1998 as a resource to collect, characterize, and to release for transplantation cord blood units (CBU) from families affected by malignant and non-malignant disorders. Currently, 1686 CBUs from 1587 families have been collected among referrals from all 50 US States. The categories of participation include malignant disorders (50%), sickle cell disease (29%), thalassemia (6%), and other hereditary or rare hematological conditions (15%). The mean cell volume collected was 102 ml (range, 31–284) with a mean total nucleated cell count (TNC) of 9.5 x 108 (range, 0.6–53.6) and mean CD 34+ cell count of 3.6 x 106 (range, 0.1–88.1). The post-thaw viability of CBU released for transplantation was 94.4 % (SD ±8.7%) and only 4.4% of CBUs were not processed due to having inadequate volume.
To date, 54 children have been treated by sibling donor cord blood transplantation (CBT), 38 using the cord blood unit as the sole source of stem cells. There was a very high rate of CBU utilization, particularly among thalassemia families where 18 of 105 (17%) of CBUs collected have been released for CBT. CBT recipients had hematological malignancies (N=20), thalassemia major (N=18), sickle cell anemia (N=8) or other non-malignant disorders (N=8), and all but 6 received HLA-identical allografts. The median total nucleated (TNC) and CD34+ cell dose was 3.1 x 107 TNC/kg and 0.7 x 105/kg recipient weight, respectively. The median time to ANC >500 and platelet >20,000/mm3 was 22 and 45 days, respectively. One of 52 (2%) evaluable patients had graft failure accompanied by autologous reconstitution. With a median follow-up of 9.6 (range, 0.1 – 94) months, 45 of 54 (83%) patients survive, and 9 patients (17%) died of relapse (N=6) or transplant-related causes (N=3) after CBT. Among the hemoglobinopathy patients, 23 of 26 (88%) survive, and 22 (85%) survive disease-free. Overall, the Kaplan-Meier probabilities of survival and event-free survival after sibling CBT are 79% and 72%, respectively.
These results confirm the feasibility of remote site, directed donor cord blood collection and subsequent transplantation for hematological disorders. The ability to combine cord blood collections with a marrow harvest from the sibling donor effectively reduced the incidence of graft rejection when CBU cell doses were judged to be insufficient. Transplantation of sibling CBUs in lieu of bone marrow may be particularly advantageous when there is urgency for transplantation and in non-malignant disorders where graft-versus-host disease, in particular, has a negative impact upon outcome.
Outcome and Immune Reconstitution Following T Cell Depleted Nonmyeloablative Allogeneic Transplantation Using Matched Donors.
Authors: Rizzieri David A. , Koh Liang Piu , Long Gwynn D. , Gasparetto Cristina , Gong Jerald Z. , Lagoo Anand S. , Niedzwiecki Donna , Sullivan Keith M. , Chute John , Horwitz Mitchell , Ashley Morris , Chao Nelson J. 
Published: October 2019

Abstract
To minimize toxicity and monitor immune recovery without interference of long term use of immunosuppressive agents, we have investigated T-cell depleted, nonmyeloablative allogeneic therapy using matched family member donors.
Methods: Seventy five patients who were not candidates for ablative therapy due to age or comorbid diseases received fludarabine 30mg/sq m and cyclophosphamide 500mg/sq m IV qd x 4 with alemtuzumab 20mg IV qd x 5 followed by stem cell infusion. No other post transplant immunomodulation was provided.
Results: Patient diagnoses included lymphoma/myeloma (20), leukemias/MDS (30), myelofibrosis/aplasia (7) and metastatic solid tumours (18). The median age was 50 (range 18–17) with a median follow up of 18 months. The median CD34+ cell dose collected was 13.4 x 106/kg. Engraftment occurred in 100% of patients with a median of 97% donor cells responsible for patient hematopoiesis by 3 months. Forty six also had a DLI (range 106–108 CD3+ cells/kg). Overall, Grade III–IV acute GVHD occurred in only 5/75 (7%) and 18 (29%) had grade II–IV. Four patients developed chronic GVHD. The transplant regimen was well tolerated with 4% 100 day treatment related mortality. In those with hematologic malignancies, only 7% started in remission, though 45 (60%) attained a CR. The most common cause of death in this group was progressive disease (28%), followed by infections (5%), and GVHD (5%). A subgroup of 21 of these older, more infirm patients who had high risk AML in 1–2nd CR or PR or ≥2 chronic stable phase CML, partially responding lymphoma, or severe myelofibrosis have been followed for at least 1 year. At 1 year, 13/21 (62%) remain alive and in continuing remission. Phenotypic analysis of lymphocyte subsets (measured by flow) revealed recovery at 6 months.
Figure Figure
T cell VBeta family recovery (spectratype analysis using PCR) revealed robust recovery by 6 months as well (first figure pre and second is 6 months post transplant), despite T depletion.
Figure Figure
TRECs analysis reveals little, if any, recovery in these patients for at least 12–18 months.
Conclusions: Nonablative allogeneic transplantation using alemtuzumab for T cell depletion results in reliable engraftment with little acute GVHD or TRM. Immune recovery assays reveal that despite T cell purging, T cell subset and VBeta families have significant recovery by 6 months and TRECS results show the recovery is primarily from peripheral expansion of transplanted donor cells, not education of new T cells. These data possibly reflect the advantage of low dose donor lymphocyte boosts without planned long term use of immunosuppressive agents. The future challenge will to be to develop strategies to further improve immune recovery in this setting.
Mode of Action of Rituximab in Chronic Lymphocytic Leukaemia; Activation of Tis11b, an Inducer of mRNA Instability, and Induction of Apoptosis.
Authors: Baou Maria , Murphy John , Jewell Andrew P. 
Published: October 2019

Abstract
Rituximab is a chimeric anti-CD20 monoclonal antibody that has been used successfully in the treatment of Non Hodgkin’s Lymphoma or patients with Chronic Lymphocytic Leukaemia (CLL). The mechanisms of action of Rituximab are not fully understood although antibody dependent cell mediated cytotoxicity and complement dependent cytoxicity have been shown to be important. An alternative mechanism is the induction of apoptosis through activation of pathways mediated through CD20. CD20 is involved in many cellular processes including proliferation, activation, differentiation and apoptosis. We have found that treatment of CLL cells with 20 μ g/ml Rituximab cross-linked with a secondary antibody reduced cell viability from 84±8% (in unstimulated cells) to 51.50±10% after 48h of cultivation by the Annexin/PI method. Using inhibitors specific for p38, JNK and ERK pathways, we found that inhibition of p38 inhibits the induction of apoptosis by crosslinked Rituximab. Rituximab has been reported to inhibit this pathway andlead to down regulation of bcl-2 expression in AIDS related lymphoma cells. However the mechanism is unclear. One mechanism by which many genes involved in apoptosis are regulated is through induction of mRNA instability through induction of Tis11 family genes. The Tis11 family (Tis11, Tis11b/Berg36 and Tis11d) bind to AU Rich elements present in several mRNA (eg bcl-2, TNF) and cause their degradation. We found that Tis11b/Berg36 is strongly induced by crosslinked Rituximab. Tis11d was weakly induced while Tis11 remained unchanged after treatment. Furthermore we found that induction of Tis11b/Berg36 by Rituximab is partly regulated through the p38 pathway since inhibition of this pathway resulted partial or complete inhibition of Tis11b/Berg36 induction. This suggests that Tis11b/Berg36 may mediate the induction of apoptosis by Rituximab through the degradation of proteins involved in apoptosis that contain AU Rich elements, disrupting autocrine cytokine feedback mechanisms and down regulating bcl-2.
Association of a Novel Regulatory Polymorphism (-938C>A) in the BCL2 Gene with Disease Progression in Chronic Lymphocytic Leukemia.
Authors: Nueckel Holger , Frey Ulrich H. , Sellmann Ludger , Duerig Jan , Duehrsen Ulrich , Siffert Winfried 
Published: October 2019

Abstract
Bcl-2 plays a key role in the regulation of apoptosis. We investigated the role of a novel regulatory single nucleotide polymorphism in the BCL2 gene (−938C>A) in B-CLL. Bcl-2 protein expression in B-cells from CLL patients carrying the −938 AA genotype was significantly increased compared to AC and CC genotypes. Moreover, the −938C- and A-alleles differed with regard to the binding of nuclear proteins.
Genotype distributions between 123 CLL patients (42 AA, 55 AC, 26 CC) and in 109 age-matched healthy controls (38 AA, 55 AC, 16 CC) were not significantly different suggesting that this polymorphism does not increase the susceptibility for B-CLL. However, time from first diagnosis to initiation of chemotherapy was significantly shorter in patients with −938AA genotype (38 months) compared to AC/CC genotypes (120 months; p=0.0077), especially in CD38 and ZAP-70 negative CLL patients. Multivariate Cox regression identified the BCL2 −938AA genotype as an independent prognostic factor for progression-free survival (hazard ratio, HR, 1.9; p=0.034) together with disease stage at diagnosis (HR 1.6; p=0.018) and ZAP-70 status (HR 2.9; p=0.001). Furthermore, the BCL2 −938C>A polymorphism in combination with the two common prognostic factors CD38 and ZAP70 status provides additional prognostic information beyond that obtained from single or double marker analysis. To the best of our knowledge this is the first report demonstrating the influence of a novel regulatory polymorphism in the BCL2 gene upon the progression of B-CLL, which, of course, requires confirmation from independent studies. On the other hand, the results presented here appear plausible with regard to differences on the protein level depending on genotype and the known impact of Bcl-2 on disease progression. Thus, our report exceeds other genetic association studies in that a mechanistic link between genotype and observed phenotype can be proposed.
In conclusion, carriage of the BCL2 −938AA genotype is a novel unfavorable genetic marker in patients with B-CLL associated with increased Bcl-2 expression.