bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Glycogensynthase Kinase 3b (GSK-3β) Is Involved in Leukemic Transformation
Authors: Bäsecke Jörg , Lehmann Anneke , Bergholz Ulla , McCubrey James , Trümper Lorenz , Stocking Carol , Horn Stefan 
Published: April 2020

Abstract
Purpose: Signal transduction pathways, such as the PI3K/AKT cascade, are frequently activated in acute myeloid leukemia and stimulate the proliferation and survival of leukemic cells. Mutations in upstream genes such as Class III receptor tyrosine kinases are frequent but not exclusive causes of that activation. An important downstream target of PI3K/AKT is the key regulator GSK-3β. It controls anti-apoptotic genes such as NF-kappaB and Cyclin D1, is involved in wnt-pathway activation and drug resistance of leukemic cells. Deregulated signaling by GSK-3β occurs through inhibition by AKT-mediated phosphorylation. We have observed that constitutive phosphorylation of GSK-3β occurred in hematopoietic cells with pro-leukemogenic PI3K-mutations. We wanted to evaluate the relevance of GSK-3β inactivation in the transformation process of hematopoietic cells.
Methods and Results: We used an in vitro factor-independent growth assay, GSK-3b inhibitors (Lithium, BIO) and established a second hit model using retroviral gene transfer of the weak oncogene Bcl XL. Signaling cascades were analysed by western blot. We demonstrate that inactivation of GSK-3b alone was not sufficient to induce factor-independent growth in IL-3 dependent early hematopoietic cells (Ba/F3). Induction of apoptosis upon growth factor withdrawal was reduced, but not prevented, in the presence of GSK-3b inhibitors, leading to a delayed Caspase 3 activation, PARP cleavage and DNA fragmentation. Overexpression of Bcl-XL also did not result in a prevention of apoptosis. GSK-3b inhibition in synergy with Bcl-XL overexpression resulted in the establishement of several growth factor-independent cell lines, which were characterized by the activation of multiple signaling cascades including AKT, MAPK, STAT5, but not STAT3. Also Cyclin D1 was overexpressed in contrast to other cyclines (D2, D3) which are no substrates of GSK-3b.
Conclusions: Our data show that GSK-3β is part of the apoptotic response to growth factor withdrawal and suggest that GSK-3β is causaly involved in the transformation process of hematopoietic cells and seems to have an synergistic role in addition to other pro-leukemic mutations.
Histone Deacetylase 11 (HDAC11) as a Novel Molecular Target in Mantle Cell Lymphoma
Authors: Villagra Alejandro v , Maurin Michelle m , Cheng Fengdong , Wang Hongwei , Dessureault Sophie , Seto Ed , Tao Jianguo , Wright Ken , Pinilla Javier , Sotomayor Eduardo M. 
Published: April 2020

Abstract
Mantle Cell Lymphoma (MCL) is a B-cell Non-Hodgkin’s Lymphoma characterized by the translocation t(11;14)(q13;32) and a consequent Cyclin D1 over-expression. The clinical evolution of MCL is very aggressive, and currently there is no standard of care for this malignancy.
In spite of the known association of Histone Deacetylases (HDACs) and solid malignancy growth, little is known regarding the role of these enzymes in MCL. We recently found newest member of the HDAC family, HDAC11, a molecule with an unknown prior physiological role, is a strong transcriptional repressor of gene expression1. Interestingly, we have found a significant over-expression of HDAC11 in murine as well as in human MCL cell lines. In primary cells from patients with MCL, we observed that higher levels of HDAC11 are associated with a more aggressive disease, while low expressions of HDAC11 are seen in patients with the indolent variant of MCL. Microarray analysis of cells in which HDAC11 expression was silenced revealed significant down regulation of specific genes involved in the control of apoptosis, cell cycle, methylation and B-cell survival. Critical genes regulated by HDAC11 include IL-10 and IL-6, cytokines known to be survival factors for malignant B-cells. In addition, we have unveiled a previously unknown association between HDAC11, cyclin D1, and Cyclin Dependent Kinase Inhibitors (CDKI), molecules that play a central role in the pathogenesis of MCL.
Potential Synergism of Green Tea and Aspirin as a Possible Cause of Bleeding in a Diabetic
Authors: Munro Roger C , Wakeman Lisa J , Al-Ismail Saad 
Published: April 2020

Abstract
A 58 year old lady with Metabolic Syndrome of 10 years duration presented with post-menopausal PV bleeding, haematuria, occasional epistaxis and ecchymoses. Prescribed medication which had remained unchanged for the preceding two years included daily doses (mgms) of Aspirin (150), Atenelol (50), Metformin (500 × 3), Bendroflumethiazide (2.5), Losartan (50) and Simvastatin (20). Intravenous urogram, cystoscopy, cytological examination of urine sediment, hysteroscopy, a cervical scan and endometrial biopsies showed neither evidence of overt pathology nor any physiological indication for the cause of haematuria or PV bleeding. Tests for urinary infection were negative. Apart from the raised blood glucose (9.1: NR 3.3 – 6.0 mmol/L), the biochemistry profile including liver enzymes, coagulation profile and blood count were normal (Platelets = 265 × 109/L). Bleeding episodes were observed after commencement of a daily intake of 7–8 cups of green tea for a period of six months. Green tea intake was self-instigated in response to reported amelioration of risk factors associated with Metabolic Syndrome (reduction in LDL cholesterol and serum triglyceride levels; elevation of protective HDL; potent antioxidant activity; ACE inhibition and promotion of glucose metabolism). Hot water extract of green tea specifically inhibits platelet adhesion and lowers sub-maximal platelet aggregation and prolongs the lag time in a dose-dependent manner. Previous fractionation studies of these hot water extracts, has revealed that the tea catechins (tannins) actively inhibit thromboxane A2 production and that ester-type catechins are more effective than free-type catechins. One of the ester-type catechins, epigallocatechin gallate (EGCG), suppresses thrombin and collagen-induced platelet aggregation completely at a concentration of 0.2 mg/ml. EGCG also inhibits aggregation by a mechanism which differs from that of aspirin by inhibiting platelet activating factor (PAF). The IC50 values of EGCG and aspirin indicate that their potencies are comparable. Bleeding symptoms ceased two weeks after the patient stopped drinking green tea. Our assumption of the causal effect of green tea on anomalous bleeding in this patient needs to be confirmed by structured platelet function tests in both aspirinised and non-aspirinised patients. Since inhibition of cyclo-oxygenase is an additional anti-thrombotic property of aspirin which differs from that of green tea, diabetic patients taking prophylactic low-dose aspirin should continue to do so and potentially beneficial ingestion of green tea should not be considered without consultation with an appropriate health professional in view of its synergistic potential on the effect of aspirin and the associated haemorrhagic risks.
Willebrand in the Netherlands: WiN Study. Background and Study Design
Authors: De Wee Eva M. , Mauser-Bunschoten Eveline P. , Van der Bom Anske G. , Eikenboom Jeroen C.J. , Fijn van Draat Karin J. , De Goede-Bolder Arja , Meijer Karina , Nováková Irena , Plug Iris , Leebeek Frank W.G. 
Published: April 2020

Abstract
The clinical presentation of patients with Willebrand Disease (VWD) is highly variable, and not always predictable upon laboratory measurements. The mechanism of this clinical variability is unknown. Also the impact of VWD on the quality of life is highly variable between patients.
This ongoing study aims to register and investigate all patients in the Netherlands with moderate and severe von Willebrand Disease to assess clinical presentation, treatment and related complications of treatment. One of the aims is to assess which factors influence the bleeding phenotype. Another goal is to investigate the influence of von Willebrand Disease on quality of life.
We have identified 1071 eligible patients. All patients are known at 12 hemophilia treatment centers. Inclusion criteria are moderate or severe VWD defined as; VWF antigen or activity ≤ 30% and/or FVIII:C ≤ 40%. The patients will be sent a questionnaire and a blood sample will be obtained for plasma and DNA.
The characteristics of the patients are the following: median age is 34 years (range 0–86) in males and 42 years (range 1–92) in females. 192 children are included (&lt;18 years). Sixty-one percent is female. In this cohort 59% has type 1, 30% has type 2, 4,5% has type 3, 0.5% is compound heterozygote and 6% is non-specified. Among type 2 we identified 50% 2A, 26% 2B, 10% 2M, 6% 2N, and 8% non-specified. In this cohort 45% has blood group 0, 29% non-0 and in 26% this is unknown. The VWF:Ag, VWF:Act (CB or RCo) and FVIII:C levels are mentioned in table 1.
Conclusion: The WiN study has identified 1071 moderate to severe VWD patients in the Netherlands who will form the basis for further research that will result in more insight into the clinical features and quality of life in VWD.
Table 1:
VWF:Ag VWF:Act* FVIII:C * VWF:CB of VWF:RCo &lt;0.10 IU/mL 91 (9%) 246 (23%) 62 (6%) 0.10 – 0.19 IU/mL 157 (15%) 368 (34%) 63 (6%) 0.20 – 0.29 IU/mL 287 (27%) 332 (31%) 137 (13%) &gt;0.30 IU/mL 527 (49%) 119 (11%) 799 (75%) Missing data 9 (1%) 6 (1%) 10 (1%)
Rituximab for the Treatment of High Titer Acquired Factor VIII Inhibitor with Long Term Follow up
Authors: Sreenivasappa Shylendra B , Telfer Margaret Clare 
Published: April 2020

Abstract
Acquired factor VIII inhibitors are auto antibodies directed against factor VIII (FVIII) and have a reported incidence of 0.2–1.0 per million individuals per year. It is a rare cause of serious bleeding associated with high mortality. Individuals with high titer antibodies (&gt; 100 Bethesda units (BU)), often have difficulty achieving a complete sustained remission although different strategies have been advocated for these patients. Rituximab has been demonstrated to show efficacy in patients with acquired hemophilia. In this study we retrospectively studied three patients with high titer acquired FVIII inhibitors with long term follow up. Two of the patients were resistant to steroids and Intravenous Immunoglobulin, one patient had received steroids and Cyclophosphamide prior to rituximab therapy. Patients ages were 36, 45 and 48 years, two were women and one male. One of the women has a h/o pregnancy about 8 months ago and other two were thought to have idiopathic inhibitors. The patient’s inhibitor titers ranged from 124–800 BU at the time of treatment with Rituximab. All received 4 weekly infusion of rituximab at 375mg/m2. No significant treatment related complication was noted. One patient did not respond to treatment. Two patients had a decline in inhibitor titers. One patient had a gradual drop in titer from 124 BU to 0.00 BU over 18 months. The other responder had a drop in titer from 800 BU to 41 BU in 16 months and then relapsed with significant bleeding and anti factor 8 titers of 700 BU and was retreated with rituximab and had a gradual drop in inhibitor titers to 50 BU over 8 months. The two patients have been followed for 28 and 30 months respectively. The non responder continues to have high tires of antibodies and has been followed for 25 months.
We conclude that in patients with high titer of acquired inhibitors rituximab alone may produce a drop in inhibitor titer but is not sufficient to achieve a complete response. Rituximab in combination with other therapies or novel agents may provide better results,
Altered Immune Responses in Patients with Chuvash Polycythemia
Authors: Niu Xiaomei , Miasnikova Galina Y , Sergueeva Adelina I , Okhotin Daniel J , Nouraie Mehdi , Ammosova Tatiana , Nekhai Sergei , Gordeuk Victor R. 
Published: April 2020

Abstract
Chuvash polycythemia results from a homozygous 598 C&gt;T mutation in exon 3 of the von Hippel-Lindau (VHL) gene. This disrupts the normoxia pathway for degrading hypoxia inducible factor (HIF)-1a and HIF-2a causing altered expression of HIF-1 and HIF-2 regulated genes. As hypoxia induces expression of pro-inflammatory cytokines, we hypothesized that there might be an elevation of Th1 cytokines in the setting of Chuvash polycythemia. Concentrations of Th1 (interleukins-2 and 12, interferon-g, GM-CSF, tumor necrosis factor-a) and Th2 cytokines (interleukins-4, 5, 10 and 13) were measured in plasma using the Bio-Plex multiplex suspension array system. Concentrations of all of these cytokines were elevated in patients with Chuvash polycythemia 598C&gt;T homozygous compared to the control wild type participants. In parallel, peripheral blood concentrations of CD3 positive T-helper cells and CD-4 positive T-helper cells were lower in patients with Chuvash polycythemia compared to controls. The ratios of interferon-g and tumor necrosis factor-a to interleukin-10 did not differ by genotype. Thus, although the upregulated hypoxic response in Chuvash polycythemia is associated with increased plasma products of the Th1 and Th2 pathways, the peripheral blood concentrations of T-helper cells are reduced, perhaps as part of a feed-back mechanism, and the balance between the two pathways may be preserved. Such countervailing responses to unregulated hypoxia sensing may explain why patients with Chuvash polycythemia do not present clinically with immunologic disorders.
Impaired Immune Reconstitution after HLA-Mismatched Transplantation Due to Antigen Specific T-Cell Recognition Restricted by the Disparate Haplotype
Authors: Amir Avital L. , Heiden Pim L.J. , Falkenburg J.H. Frederik , Heemskerk Mirjam H.M. 
Published: April 2020

Abstract
The applicability of HLA matched hematopoietic stem cell transplantation (SCT) is limited by the lack of HLA matched donors. In contrast, an HLA haploidentical donor can be found for almost all patients. Posttransplantation immunodeficiency leading to high morbidity and mortality from viral infections is a major barrier to the broader application of haploidentical SCT. In HLA mismatched SCT profound T cell depletion is necessary to prevent GVHD, but also leads to slower T-cell reconstitution. Vigorous T-cell depletion, however, does not fully explain the increased immunodeficiency. Despite a gradual normalization of absolute T-cell and total lymphocyte number after haploidentical SCT, the immunodeficiency may persist for years. Additionally, an increase in the number of mismatched HLA alleles between patient and donor leads to more profound post transplantation immunodeficiency. We hypothesized that another explanation for the long lasting immunodeficiency is T-cell antigen recognition via the haplotype disparate between patient and donor. T-cells recognizing viral epitopes in the donor non-shared HLA molecule will attack virus infected hematopoietic cells of donor origin but not virus infected non-hematopoietic cells of patient origin. To investigate the role of T-cell disparate haplotype restriction in posttransplantation immunodeficiency, we characterized the immune responses against CMV in two patients at various time points after haploidentical and mismatched SCT. The first patient was HLA-A2 negative and transplanted with SCT of an HLA-A2 positive haploidentical donor. After the SCT he suffered from life threatening CMV reactivations for almost two years. Using tetramers composed of the HLA-A2 molecule and CMV-pp65-NLV epitope, we determined that in this patient CMV specific T-cells restricted to the donor HLA-A2 were present from 6 months after the SCT and remained present during the two year period of recurrent CMV infections. Since the HLA-A2 allele was not shared between patient and donor, these results indicate that T cells restricted by non-shared donor-HLA were unable to protect against infections. The second patient transplanted with an HLA mismatched SCT, suffered from CMV reactivations only in the first 6 months after SCT. Already 3 months after SCT CMV specific T cells restricted to the shared HLA-B8 allele were present in high concentrations, whereas the CMV specific T-cells restricted to the non-shared donor HLA-A1 allele disappeared in time. Our results suggest that for an effective immune response the T cells need to recognize viral epitopes restricted to the HLA shared between patient and donor and that impaired immune reconstitution after haploidentical SCT is, in part, the result of antigen specific T cell responses restricted to the disparate haplotype.
Exploiting Cytokine Secretion to Rapidly Produce Multivirus-Specific T Cells for Adoptive Immunotherapy
Authors: Fujita Yuriko , Leen Ann M. , Sun Jiali , Nakazawa Yozo , Yvon Eric , Heslop Helen E. , Brenner Malcolm K. , Rooney Cliona M. 
Published: April 2020

Abstract
Viral infections remain a major cause of morbidity and mortality after hematopoietic stem cell transplantation (HSCT), and conventional small-molecule therapeutics often have modest benefit, high cost and adverse effects. Adoptive transfer of donor-derived, virus-specific T cells specific for cytomegalovirus (CMV), Epstein-Barr virus (EBV) and adenovirus (AdV) proved feasible and safe after HSCT, and reconstituted immunity to all 3 viruses. Current protocols to generate multivirus-specific T cells are lengthy, taking up to 12 weeks. Since viral infections often occur &lt;30 days after HSCT, more rapid production is required. The IFN-γ capture (Miltenyi) and tetramer methods can select antigenspecific T cells directly from peripheral blood and have been used clinically to prevent infections with AdV and CMV respectively. Tetramer selection is limited to certain HLA class I molecules with known epitope specificities. By contrast, the IFN-γ capture assay can be used to select multi-epitope and multi-antigen, CD4+ and CD8+ T cells from any donor, irrespective of HLA type. We now describe a modified rapid selection method for production and characterization of CD4+ and CD8+ T cells specific for CMV, EBV and AdV in a single infusate using the IFN-γ capture assay. We could manufacture a specific product from small volumes of peripheral blood over a 48 hour time period. These cells are suited for infusion into immunocompromised patients. Activated PBMCs were transduced either with Ad5f35pp65 vector or Ad5f35LMP2 vector and then combined. After overnight culture, IFN-γ-secreting antigen-specific T cells were captured. The number of selected cells ranged from 0.06 ×106 to 0.36 ×106 cells (mean: 0.14 ± 0.06 ×106; n=5) per 20 × 106 PBMCs.Importantly, we demonstrate that the selected cells lack alloreactivity, indicating that the infused cells should be safe in vivo. By expanding a small fraction of the selected product we could characterize the phenotype, specificity and function of the infused product, to allow subsequent comparison with clinical outcome. All products were able to secrete IFN-γ in response to each antigen and kill CMV, EBV and AdV antigen-expressing targets. These results show that we can successfully generate trivirus-specific T cells for infusion within 3 days without sacrificing the product characterization required to correlate product with clinical outcome.
Immunopathology of ATG Prior to Bone Marrow Transplantation in Mouse Model
Authors: Sadeghi Behnam , Hashmi Sulaiman Al , Hassan Zuzana , Rozell Bjorn , Abedi-Valugerdi Manuchehr , Hassan Moustapha 
Published: April 2020

Abstract
Introduction: Polyclonal anti-thymocyte globulin (ATG) is mainly used in hematopoietic stem cell transplantation, as a prevention for graft rejection as well as for GVHD. Immunosuppressive properties of ATG have been considered primarily from the depletion of peripheral lymphocytes. However direct or indirect effects of ATG on other immune components still is controversial. In the present study we evaluated the immunopathologic effects of ATG on immune system in a mouse model.
Methods Ten to fourteen weeks old BALB/c and C57BL/6 mice, were injected intra- peritoneally or intravenously with different doses of ATG (0.2mg/kg -25mg/kg) at three dosage schedules. Considering bone marrow transplantation as day 0, ATG were given at days -10, -7 and -5 or -7, -5 and -1 or -5, -3 and -1. At day 0, mice were killed; spleen (SP), bone marrow (BM), lymph nodes (LN) and thymus were removed and analyzed for T-, B-, DC, NK-, T-reg and myeloid cells.
To evaluate the efficacy of immunosuppressive effect of ATG, a group of BALB/c mice were conditioned using busulfan (Bu) and ATG and compared to a control group of BALB/c conditioned with Bu (80mg/kg) followed by cyclophosphamide (200mg/kg). Both groups transplanted with BM and spleen cells from C57BL/6 and followed for engraftment and/or GVHD.
Results The administration of ATG (0.2- 4.5mg/kg) has resulted in an increase in spleen cellularity while in lymph node and thymus a decreased cellularity was observed. We have found that injecting 4.5mg/kg of ATG at day -7, -5 and -3 significantly decreased T-cell population in spleen and LN compare to Bu-Cy conditioning. The ratio CD4/CD8 increased after ATG treatment showing that CD8 cells are six-fold more sensitive to ATG treatment compared to CD4 lymphocyte. Interestingly T-reg cell population increase after ATG at day 0, however, the increment was negatively correlated with the administration time and positively correlated with the dose.
ATG treatment has resulted in an increase in B-cell population by two- and three- fold in spleen and lymph nodes, respectively. Moreover, 1.2- to 3.5-fold increase in DCs, NK and myeloid cells was observed in SP and LN. Thymus cellularity and cell phenotype was less affected while BM cellularity was not affected by ATG treatment. No differences in the ATG effect on cellularity or cell phenotype between IP and IV route was observed.
Despite the high immunosuppressive effect observed in T-cell population compared to that seen in Bu-Cy conditioning, no chimerism was observed when Cy was substituted with ATG (4.5 – 10 mg/kg).
Conclusion No donor chimerism could be obtained using ATG as a single agent up to 25mg/kg. ATG dose and the administration time are important factors affecting the repopulation of residual T-cells in spleen and lymph node that have to be considered in bone marrow transplantation setting.
The Study on Influence of T Cell Ratios Between Grafts to Recipients on Gvhd in HLA- Haploidentical Allogeneic Peripheral Blood Stem Cells Transplantation
Authors: Liang Hui , Deng Xiao-hui , Weng Xia 
Published: April 2020

Abstract
Objective To study the influence of T cell ratios between grafts to recipients on GVHD in HLA- haploidentical allogeneic peripheral blood stem cells transplantation ( PBSCT ).
Methods:
Animal experiment: We set up rat allo-BMT model. Male SD rats were transplanted with female Wistar rats.Recipient rats of experiment group were transplanted with the CD3+ T cells of female Wistar rats at different graft-receptor T cell ratios, 1:1 (group A1), 2:1 ( group A2), 4:1 ( group A3) respectively. Control group (group B) were infused with grafts consisting of 3×107 bone marrow cells by classical method. Observe the incidence and severity of GVHD of different groups. Clinical trial: 6 cases of HLA-haploidentical allogeneic PBSCT. 4 patients were transplantated with purified allo-CD34+ cells.
Next day, adjusted graft-receptor T cell ratios to 2:1 by infusing donor CD3+ T cells. Another 2 patient were treated with classical allo-PBSCT.
Results
Animal experiment: The severity of GVHD in group A1, A2 were lighter compared with group A3,B. The difference is significant among group A1,A2 and A3. Clinical trial: An ultra-acute severe GVHD occurred in 2 patients who were transplanted with classical method.
They died from interstitial pneumonia secondary to PBSCT. The remaining 4 cases only presented mild aGVHD.Among these 4 cases, 3 patients had DFS (disease free survival) over 3 years, 1 patient died from polyradiculitis on +155d.
Conclusion: On the basis of our animal experiment, the appropriate graft-receptor T cell ratios would be 1:1 or 2:1. If we applied this ratio to clinical transplantation, especially to part HLA-related and HLA-haploidentical transplantation, we could obtained a better GVL effect and a lighter GVHD.