bluuebunny/crossref_metadata_2025_split huggingface dataset:
Study on Mechanisms of Telomerase Regulation in Arsenic Trioxide Inducing Apoptosis in Myelodysplasia Cell Line.
Authors: Tong Hongyan , Jin Jie , Xu Weilai , Qian Wenbin , Lin Maofang 
Published: October 2019

Abstract
The telomerase activity can be down regulated by arsenic trioxide (As2O3), which is regarded as an apoptotic induction agent, is confirmed in many kinds of tumor cells. To investigate the mechanisms of telomerase regulation and to explore the correlation of As2O3 inducing apoptosis and telomerase regulation in MUTZ-1 cells, which are established as a high-risk myelodysplasia Cell line that derived from a MDS patient (FAB subtype refractory anemia with excess of blasts), a quantitative assessment of the telomerase activity by TRAP-ELISA and detection of the expression levels of hTERT, TRF1 (TTAGGG repeat binding factor 1), TRF2 (TTAGGG repeat binding factor 2), bcl-2, bax mRNA were performed, together with the assessment of the apoptosis by means of translocation of phosphatidylserine (PS) through flow cytometry assay. The results indicated that a typical apoptotic cell group distribution of DNA content was represented in the MUTZ-1 cells after being exposed to As2O3 at the range of concentration from 1μmol/L to 8μmol/L in a dose-dependent manner (r=0.736, P<0.001) and time-dependent manner (r=0.674, p<0.05), and the telomerase activity was down-regulated in a time-dependent manner (r=−0.976,P=0.024), and the expression level of hTERT mRNA in MUTZ-1 cells was represented in a dose-dependent manner (r=−0.892,P=0.042) and time-dependent manner (r=−1.000,P=0.04), after the cells were treated by As2O3 at the dosage as above. It was showed that a significant correlation between the decreased telomerase activity and the increased percentage of apoptotic cells in the treated cells (r=0.938,P=0.018), and there was a strong relationship between the telomerase activity and the mRNA expression of hTERT gene (r=0.783,P=0.022). However, As2O3 has no obvious effect on the expression level of TRF1 mRNA and TRF2 mRNA, which were regarded as two telomere-binding proteins. Further findings indicated that the inhibition of telomerase activity in MUTZ-1 cells was accompanied with down-regulated mRNA expression of bcl-2 gene (densitometry readings: 0.255±0.017 vs 0.466±0.069, P<0.05) and decreased ration of bcl-2/bax (densitometry reading ratios: 0.890±0.083 vs 0.546±0.014, P<0.05) at the dosage of 4μmol/L for 24 hours. These observations suggest that the apoptosis induced by As2O3 on MUTZ- 1 cells might be mediated through the inhibition of telomerase activity regulated by expression of hTERT gene, which implies that may be one of the mechanisms of As2O3 inducing apoptosis in MUTZ-1 cells.
Lysophosphatidic Acid (LPA) Protects Primary Chronic Lymphocytic Leukemia (CLL) Cells from Apoptosis: Role for LPA as a Survival Factor in CLL.
Authors: Gibson Spencer B. , Hu Xiaojie , Haney Neil , Kabore Albert F. , Johnston James B. 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of CD19+/CD5+ lymphocytes, and this is achieved primarily through a block in apoptosis. The mechanisms underlying this defect are not fully understood. Several proteins have been shown to protect CLL cells from apoptosis and one of these is albumin that solublizes lipids in the plasma. A lipid found in plasma, lysophosphatidic acid (LPA), protects epithelial and fibroblast cells from apoptosis. We investigated whether LPA could be a survival factor in CLL. Herein, we demonstrate that LPA effectively protects the B cell line BJAB and the CLL-like cell line I-83 from etoposide, fludarabine, and chlorambucil-induced apoptosis. In primary CLL cells, plasma from either healthy or CLL patients significantly reduces spontaneous and drug-induced apoptosis. However, delipidation of the plasma reduces its protective effect. In addition, LPA protects primary CLL cells but not healthy lymphocytes from apoptosis. By western blotting, the LPA receptor 1 (LPA1) expression is increased in primary CLL cells compared to normal lymphocytes. Treatment of primary CLL cells with the LPA receptor antagonist diacylglycerol pyrophosphate (DGPP) reverses the protective effect of LPA against apoptosis. Over expression of the LPA1 receptor protects cells from apoptosis and downregulation of the receptor blocks LPA mediated protection against spontaneous apoptosis. The protective effect of LPA is inhibited by blocking activation of the PI-3K/AKT signaling pathway. These results indicate that LPA is a survival factor in primary CLL cells and that drugs targeting the LPA receptors might be an effective therapy for this disease.
Monoclonal Gammopathy of Undertermined Significance (MGUS): Clinical Predictors of Malignant Transformation in 434 Patients with a Long Follow-Up from a Single Institution.
Authors: Rosiñol L. , Montoto S. , Bladé J. , Cibeira M.T. , Rozman M. , Nadal E. , Giné E. , Aymerich M. , Esteve J. , Montserrat E. 
Published: October 2019

Abstract
Background: MGUS is a common disorder characterized by the presence of a small serum M-protein in individuals with no evidence of multiple myeloma (MM), Waldenström’s macroglobulinemia (WM) or primary amyloidosis (AL). Although about one fourth of these individuals will evolve into a malignant disease with a transformation rate of 1% per year, there are not well-established predictors of outcome.
Aim: To identify predictor features of malignant transformation in a large series of patients with MGUS and prolonged follow-up.
Patients and methods: Four hundred and thirty-four patients (200 M/234 F; median age 66 years) diagnosed with MGUS in a single institution from September 1970 to January 2001 with a minimum follow-up of one year were included in the study. All patients had an M-protein size < 30g/L. Bone marrow aspirates were reviewed independently by two of the authors and the proportion of bone marrow plasma cells (BMPC) were estimated from a 500 cell-count by each observer. The median follow-up was 5.2 years (range: 1–28.8 years) with 84 patients followed for more than 10 years.
Results: The type of M-protein was IgG in 67.2% of the cases, IgA in 18.8%, IgM in 11.9%, light chain in 1% and biclonal in 1%. The light chain was kappa type in 56.4% of the patients. The median M-protein size was 15.6 g/L (<10 g/L in 10.8%, 10–20 g/L in 61.7%, and > 20g/L in 27.4%). The median percentage of BMPC in 305 reviewed samples was 4.6% (range: 0.4–25). After a median follow-up of 5.2 years, 50 patients (11.5%) have evolved into a malignant monoclonal gammopathy (44 MM, 5 WM and 1 AL). The median time to progression was 5.4 yrs (range: 1.4 – 16.9). The risk of transformation was 15.4% (95% CI: 10.5–20.3) and 34% (95% CI 22.6–45.3) and 34% (95% CI: 22.6–45.3) at 10 and 20 years, respectively. The variables associated with a higher risk of transformation were IgA-type (p=0.003), kappa light chain (p=0.009), the amount of M-protein (<15 vs >15 g/L, p=0.005) and the percentage of BMPC (<5% vs >5%, p=0.007).
Conclusions: In this series of patients with MGUS, the type (i.e. IgA or kappa) and size of M-protein (>15g/L) as well as the percentage of bone marrow plasma cells (>5%) significantly predicted malignance transformation.
Fatty Acid Synthase Inhibitor Induces Apoptosis and Inhibits Proliferation in Multiple Myeloma Cells.
Authors: Wang Weiqin , Wu Dong 
Published: October 2019

Abstract
Objective: To investigate the effect of fatty acid synthase (FAS) inhibitor (cerulenin) in multiple myeloma cells and its mechanism.
Methods: Proliferatory inhibition rate of cerulenin in multiple myeloma cells U266 and RPMI8226 was assayed by MTT, apoptosis analysis and cell DNA cycle analysis by flow cytometry (FCM).
Results: Treated with 5ug/ml~640ug/ml of cerulenin for 24h, the proliferation of U266 and RPMI8226 cells were obviously inhibited with dosage related effect at the same concentration, the inhibition rate of human skin fibroblast are all lower than 28%. When U266 and RPMI8226 are treated for 12h with 20ug/ml of cerulenin the apoptosis rate revealed by FCM are 51.3% and 48.8% respectively. Cell DNA cycles analysis show the cell DNA is blocked in G1 phase and S phase cells decreased in both of U266 and RPMI8226.
Conclution: FAS cold induce MM cell apoptosis and inhibit MM cell proliferation. FAS might be a new potential target for MM therapy.
p38 alpha MAP Kinase Inhibitor Inhibits Human Myeloma Cell Growth in Vivo.
Authors: Reddy Mamatha , Ma Jing Y. , Kerr Irene , Henjarappa Nirupama , Navas Tony , Chakravarty Jit , Dugar Sundeep , Higgins Linda S. , Protter Andrew A. , Medicherla Satyanarayana 
Published: October 2019

Abstract
The p38 MAPK signaling pathway plays an important role in different pathological conditions and specific inhibitors of p38 alpha and beta MAPK block production of major inflammatory cytokines. Human multiple myeloma (MM) is an incurable neoplasm of B cells, and MM disease progression is affected by immunoregulatory elements such as IL-6 and other cytokines. Recent literature indicates that an important function of p38 MAP kinase is the generation of signals of critical value to the control of normal and malignant hematopoiesis by cytokines and growth factors. We therefore examined the effect of a p38 alpha MAP kinase specific inhibitor, SD-282, on human myeloma cell (RPMI8266) tumor growth in immunodeficient beige-nude-xid mouse xenograft plasmacytoma model. Treatment (90 mg/kg/bid by oral gavage) was initiated in mice with palpable tumor size (~200 mm3) as judged by tumor volume. In mice with palpable tumors, 21 days of SD-282 treatment significantly reduced tumor volume. When SD-282 treatment was initiated in mice with pronounced tumor size (~800 mm3) there remained a significant reduction in tumor volume. Histological assessment at the end of dosing from animals administered SD-282 or vehicle demonstrated a significant reduction in tumor volume and number of neoformed microvessels in the SD-282 treatment group. It also significantly reduced HSP27 and p38 expressions in tumor cells.
In conclusion, we report that the p38 inhibitor inhibits human myeloma cell growth in vivo via p38 MAP kinase signaling pathway. The current study provides strong in vivo evidence supporting use of p38 inhibitor therapy in patients with multiple myeloma
Rapid Clinical Scale Isolation of CD25hiCD4+ Regulatory T Cells.
Authors: Niemand Claudia , Conrads Carina , Siemer Ramona , Assenmacher Mario 
Published: October 2019

Abstract
Several publications during the last few years have reported CD25hiCD4+ regulatory T cells (Tregs) to prevent or to reverse disease in different mouse models of experimental autoimmune encephalomyelitis (EAE), colitis, graft rejection and graft-versus-host-disease (GvHD). As mouse and human Tregs share many phenotypical and functional characteristics, Tregs could provide a promising therapeutic approach for various human autoimmune diseases and pathological alloresponses. Here we have shown that Tregs can be isolated from leukapheresis harvests by CD25 enrichment using the CliniMACS technology (n=13). By this procedure we obtained 2.32x108 (± 1.12x108, range 0.71–4.42x108) cells out of 1010 mononuclear cells with a mean purity of 52.12% (± 12.11%, range 25.48–66.61%) for CD25hiCD4+ cells. Around 90% of enriched cells were CD25+CD4+. Among contaminating CD4− cells most cells were CD25+ which were further characterized by counterstaining to be mainly CD19+ B cells and a few CD8+, CD56+ or CD123+ cells. It is possible to deplete the CD19+ or CD8+ cells by using CD19 Microbeads or CD8 Microbeads respectively with the CliniMACS Instrument before CD25 enrichment. Combined depletion of different cells, e.g. CD19+ and CD8+ cells is conceivable. Isolated cells were phenotypically and functionally characterized. The majority of the CD25hiCD4+ T cells expressed glucocorticoid-induced tumor necrosis factor receptor (GITR), CD62L and CD45RO. In addition, isolated cells were able to suppress the proliferation and activation of cocultured conventional CD4+ cells after polyclonal stimulation with anti-CD3 antibody. We conclude that the large-scale isolation of CD25hiCD4+ regulatory T cells for clinical applications (e.g. therapy of autoimmune diseases, graft rejection or GvHD) is possible by using the CliniMACS CD25 Reagent and the CliniMACS Instrument.
Development of Functional Haemophilus Influenzae Type B Antibodies after Vaccination of Autologous Stem Cell Transplant Recipients.
Authors: Van der Velden Ankie M.T. , Claessen Anke M.E. , Van Velzen-Blad Heleen , Biesma Douwe H. , Rijkers Ger T. 
Published: October 2019

Abstract
The conditioning regiments for autologous stem cell transplantation (aSCT) lead to impairment of the immune system and susceptibility to infections. Although infectious complications are more prominent in allogeneic transplantation, it is also a major source of morbidity in patients undergoing aSCT. Therefore, vaccination of patients after stem cell transplantation has been recommended. We conducted a prospective follow-up study to determine quantitative and qualitative aspects of the humoral immune response to multiple vaccinations with conjugated Haemophilus influenzae type b (Hib) vaccine in autologous stem cell transplant recipients. Sixteen patients (twelve with multiple myeloma and four with non-Hodgkin’s lymphoma) received a conjugated Hib vaccine at 6, 8 and 14 months after transplantation. Antibody titres were determined by ELISA, 21 days after each vaccination. Adequate anti-Hib antibody responses (i.e. a four fold or greater increase in antibody levels in addition to a minimal titre of 50 U/ml corresponding to 18.8 μg/ml) were achieved in 7 (44%) of the patients after one vaccination. A second and third vaccination resulted in an adequate anti-Hib antibody response in 12 (75%) and 14 (88%) patients, respectively. Functionality of anti-Hib antibodies was assessed by measurement of antibody avidity. Repeated vaccination induced maturation of antibody avidity as demonstrated by increased resistance to sodium thiocyanate (NaSCN) treatment (Figure 1; the avidity index (AI) is defined as the molarity of NaSCN at which 50% of the amount of IgG antibodies bound to the coated antigen in the absence of NaSCN has been eluded. Mean AI plus range are shown after one, two and three vaccinations.). Subsequently, opsonizing activity of antibodies was determined by a phagocytosis assay. FITC-labelled Hib was incubated with serum and added to fresh polymorphonuclear cells (PMN). Mean fluorescence intensity (MFI) was measured by flowcytometry. Anti-Hib antibodies supported phagocytosis by PMN’s after multiple vaccinations (Figure 2; MFI at different time points is shown). In conclusion, multiple vaccinations with Hib-conjugate vaccine in autologous stem cell recipients result in high antibody response rates and functional maturation of antibodies.
The Use of Reduced Intensity Allogeneic Stem Cell Transplantation (alloSCT) for Recurrence after Autologous Stem Cell Transplantation (ASCT) in Hodgkin’s Disease (HD).
Authors: Krishnan Amrita Y. , Nademanee Auayporn , Fung Henry C. , Popplewell Leslie , Rodriguez Roberto , Kadhimi Zaid Al , Kirschbaum Mark , Kogut Neil , Raubitschek Andrew , Schriber Jeffrey , Zain Jasmine , Smith Eileen , Cheng Nigel , Qian Dajun , Molina Arturo , Forman Stephen J. 
Published: October 2019

Abstract
Autologous stem cell transplantation(ASCT) can cure up to 50% of patients with relapsed or refractory HD. However, for patients who relapse post ASCT or develop post transplant myelodsyplasia (t-MDS), standard chemotherapy options offer little chance of long-term disease free survival (DFS). The use of allogeneic stem cell (alloSCT) may provide a means of control of the HD through the immune mediated effects of an allogeneic approach and also be curative for concomitant t-MDS. While traditional alloSCT in HD patients was fraught with high treatment related mortality, a reduced intensity conditioning regimen may reduce upfront mortality while still providing long term DFS.
Between 11/00 and 1/04, fourteen patients underwent reduced intensity alloSCT at the City of Hope Cancer Center. Median age at alloSCT was 31years (range18–47). Median time from HD diagnosis to alloSCT was 36mos (range 3–123). Eleven patients had relapsed post ASCT and one post syngeneic transplant. One of the pts also had t- MDS. The median time from ASCT to alloSCT was 20 mos(range1–45). Ten pts received sibling alloSCT and four unrelated donor. The conditioning regimen consisted of Fludarabine 125mg/m2+ Melphalan 140mg/m2. All patients received a Cyclosporin(CSA) based graft versus host disease (GVHD) prophylaxis regimen; nine in conjunction with Mycophenolate Mofetil and three in conjunction with Methotrexate. Seven developed acute GVHD (3-GrIII-IV). Two patients died of GVHD. There was no other significant regimen related toxicity. Median length of followup for surviving patients is 55 mos. For eleven evaluable pts, one year OS and DFS are 77% (95%CI 49–100) and 58% (95%CI 25–91) respectively (figure 1). The pt with concomitant t-MDS also achieved a hematologic remission. In conclusion, reduced intensity alloSCT may provide a salvage for heavily pretreated patients with HD that have failed ASCT and who otherwise have a poor prognosis, and it can also provide a treatment for late secondary complications of transplantation such as t-MDS.
Molecular Relapse of Acute Lymphoblastic Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation Can Be Successfully Treated with Donor Lymphocyte Infusions.
Authors: Dominietto Alida , Raiola Anna Maria , Grasso Raffaella , Garuti Anna , di Grazia Carmen , Bregante Stefania , Gualandi Francesca , Ibatici Adalberto , Lamparelli Teresa , van Lint Maria Teresa , Frassoni Francesco , Bacigalupo Andrea 
Published: October 2019

Abstract
Background. Hematologic relapse is seen in 20–40% of adult acute lymphoblastic leukemia (ALL) patients undergoing an allogeneic hematopoietic stem cell transplant (HSCT) and is usually non responsive to donor lymphocyte infusions (DLI). This has suggested a lack of graft vs leukemia effect (GvL) in ALL patients. It is currently possible to monitor minimal residual disease (MRD) post-HSCT and this may allow to identify patients at high risk of hematological relapse.
Objectives. To monitor MRD in B-ALL patients post-HSCT and to treat MRD positive patients with DLI.
Patients and Methods. MRD was evaluated on bone marrow samples using nested polymerase chain reaction (PCR) and real time PCR (RT-PCR) assay to analyze respectively IgH VDJ and BCR-ABL gene rearrangement. Molecular positivity/relapse was defined with two consecutive positive PCR assays. MRD monitoring was performed in 28 patients grafted from an HLA-identical sibling (n=19), family mismatched related donor (n=2) or matched unrelated donor (n=7). Median follow-up was 47 months (range 5–163). Sixteen patients (57%) were in early disease phase at the time of transplant.
Results. We identified 3 groups of patients. A) 14 patients (50%) had no evidence of MRD after HSCT. B) 6 (21%) had a positive MRD and received escalating dose DLI, within 60 days and C) 8 (29%) had a positive MRD, but did not received DLI, because not available or because of an early hematological relapse. Median time from HSCT to molecular relapse was 139 days (range 46–1048). The median follow up was 1385 days (144 – 4877). Median number of infused CD3+ cells was 0.6x10^7/Kg of recipient body weight (range 0.01–7). Hematological relapse was seen in 0%, 0% and 88% respectively of group A,B,C (p=0.0001) and disease-free survival (DFS) was 100% , 67% , 12% (p=0.0001).
Conclusions. This study shows that (1) MRD monitoring of ALL patients post-transplant identifies patients at high risk of hematologic relapse and (2) that treatment with DLI, on the basis of MRD positivity, significantly reduces the risk of leukemia relapse and may improve DFS. These data also confirm the existence of a GvL effect, previously shown in this disease (Weiden et.al NEJM, 300, 1068; 1979).
Drug Resistance and In Vitro Selection of Human Hematopoietic Cells Transduced with Cytidine Deaminase.
Authors: Flasshove Michael , Rattmann Ina , Lehmberg Kai , Bardenheuer Walter , Sorg Ursula R. , Seeber Siegfried , Moritz Thomas 
Published: October 2019

Abstract
Retroviral gene transfer of human cytidine deaminase into murine hematopoietic cells has been shown to confer drug resistance against cytidine analogues such as araC (1-β-D-arabinofuranosylcytosine) and gemcitabine (2′,2′-difluorodeoxycytidine). Since data for primary human hematopoietic cells are lacking so far, we transduced human CD34+ enriched umbilical cord blood cells with the retroviral vector SF91-CDD-IRES-EGFP containing the cDNA of cytidine deaminase (CDD) and EGFP as a reporter gene. The transduced cells were tested in clonogenic assays for resistance against increasing concentrations of araC or gemcitabine. Gene transfer of SF91-CDD-IRES-EGFP resulted in a transduction efficiency of 28 ± 2% (n = 4). A significant higher survival of CFU-C in the presence of araC could be demonstrated as compared with the mock control expressing EGFP only. This protective effect was improved by FACS enrichment of EGFP+ cells prior to functional analysis, and was observed for a dose range of 30 – 500 nM araC. Gemcitabine resistance was also tested with cells enriched for EGFP expression. For CFU-C a significant higher survival of colonies at concentrations of 8 and 10nM could be shown. This protection was more pronounced for CFU-GM than for BFU-E. The IC50 value increased 3fold for CFU-GM but only 1.4fold for BFU-E and 1.8fold for total CFU-C in comparison to the mock control (n = 4). In vitro selection of CDD transduced progenitor cells was investigated by incubation in the presence of 30 – 100nM araC for 4 or 8 days. Before and after culture cells were analysed for EGFP expression and tested in clonogenic assays for araC resistance. Significant selection of CDD transduced cells as measured by EGFP expression was observed for all examined concentrations. After 4 days incubation with 60nM araC clonogenic assays revealed a significant selection of progenitor cells resistant to 100 – 500nM araC. Incubation of transduced cells in 6 – 15nM gemcitabine for 4 or 8 days likewise allowed successful in vitro selection. Transplantation of CDD transduced CD34+ cells in sublethally irradiated NOD/SCID mice resulted in up to 81% engraftment of human CD45+ cells and a rate of up to 4.6% of human CD45+/EGFP+ cells after 8 weeks. These data demonstrate that retroviral gene transfer of CDD protects human hematopoietic cells from the cytotoxic drugs araC and gemcitabine and allows selection of transduced cells. Successful engraftment of CDD transduced cells in NOD/SCID repopulating cells could be observed.