bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Hes-1 (Notch-1 Effector) during “In Vitro” Normal Erythroid Differentiation.
Authors: Cappellini Maria D. , Libani Ilaria V. , Calzavara Elisabetta , Ronzoni Luisa , Chiaramonte Raffaella , Comi Paola 
Published: October 2019

Abstract
Hematopoiesis involves highly regulated proliferation and differentiation during which a small number of multipotent stem cells give rise to differentiated progenies. In several developmental systems stem cells fate is influenced by soluble molecules acting via cell-cell interaction, including those mediate by the Notch receptor family. Members of Notch family transmembrane receptors are found on primitive hematopoietic precursors, suggesting a role for Notch signaling in mammalian blood cells development. Notch signaling regulates cell fate controlling asymmetric cell division during stem/progenitor cell differentiation. A previous study on K562 cell line showed that Notch signaling inhibits erythroid/megakaryocytic development by suppressing GATA-1 activity. Furthermore there are evidences that Notch is expressed in early murine erythroid precursors. Probably Notch signaling in these uncommittted precursors may lead to enhanced survival, preserving multilineage potential. The role of Notch pathway during human adult erytropiesis has not been described. The aim of this study is to investigate the modulation of Notch activity during “in vitro” human erythropiesis. Human CD34+ from perpheral blood of normal adult subjects were differentiate “in vitro” for two weeks by the addiction of IL-3, SCF and Epo. This method of colture reproduces all stages of adult erythropoiesis. We analized the modulation of the expression of Notch-1, of its effector Hes-1 and of several erythoid specific genes, at different stages of differentiation using real time PCR. Our analysis shows that Hes-1 expression, which indicates the activation of Notch-1 pathway, is very high in the early steps of differentiation (BFU-E, CFU-E) while in the late stages rapidly decreases to undetectable levels. The Notch-1 gene expression doesn’t seem to be modulated way, but we didn’t invstigate the protein levels yet. These data suggest that Notch pathway is involved in the early stages of erythroid differentiation where it may enhance erythroid progenitors survival up to CFU-E, as hypothisized in mouse model, preventing them from apoptotic stimuli and promoting their proliferation. Involvement of Notch-1 signaling in preventing erythroid progenitors from apoptosis during erythroid differentiation could be important in some erythropoietic disorders such as b-Thalassemia syndromes or diserythropoietic anemias.
Mechanisms of Increased Reactive Oxygen Species (ROS) Generation Induced by Organic Arsenic Derivative S-Dimethylarsino-Glutathione (SGLU; ZIO-101).
Authors: Cheng Xiaodong , Taghi Manshouri , Huang Peng , Golemovic Mirna , Zingaro Ralph , Freireich Emil J. , Andreeff Michael , Kantarjian Hagop M. , Verstovsek Srdan 
Published: October 2019

Abstract
ZIO-101 is organic arsenic with similar anti-leukemia activity to arsenic trioxide (ATO) but much less toxicity. Consequently, ZIO-101 can be given at substantially higher doses than ATO and may be active in more diverse cancers than ATO. The precise anti-cancer mechanism of ZIO-101 is unknown: ZIO-101 increases intracellular production of reactive oxygen species (ROS) in dose dependent manner, resulting in apoptosis of leukemic cells. Modifying intracellular ROS levels alters the ability of ZIO-101 to induce apoptosis. We focused on the 2 most important intracellular ROS generators, mitochondria and NADPH oxidase. Cells were treated with ZIO-101 and NADPH oxidase expression levels, membrane translocation and interactions of subunits were studied. Effects of modified NADPH oxidase levels on ZIO-101-induced ROS production were studied (diphenyleneiodonium [DPI], a specific inhibitor of NADPH oxidase, and SiRNA technique were used to inhibit enzyme function, while bryostatin-1, a specific activator of NADPH oxidase, was used to increase enzyme function). Mitochondrial DNA-depleted HL60 Rho-0 cells were also used to evaluate mitochondrial input to ROS-generation. Our data show that ZIO-101 activates NADPH oxidase. A low concentration of ZIO-101 (1uM) requires 14 h to significantly increase intracellular ROS levels and kill leukemic cell. This effect is strongly inhibited by DPI-pretreatment and is also seen in P47-SiRNA transfected HL60 cells. On the other hand, this A higher dose of ZIO-101 (4uM) increases ROS levels more rapidly (2–6 h). This early increase is not inhibited by DPI-pretreatment or SiRNA transfection. A 14 h but not 2–6 h increase in ROS is detectable in Rho-0 HL60 cells. These data indicate dose-dependent mechanisms of ZIO-101 induction of ROS:
1uM ZIO-101 induces NADPH oxidase activity which results in ROS production detectable at 14 h; there is no mitochondrial component; 4uM ZIO-101 disrupts mitochondrial function resulting in an earlier increase in ROS levels and subsequent apoptosis. effect is significantly enhanced in cells that have increased enzyme function. ROS-production in Rho-0 HL60 cells is unaffected by these interventions.
A Biologically Distinctive Subgroup of Acute Myelogenous Leukemia with Characteristic Nuclear Morphology and a High Frequency of FLT3 Mutation.
Authors: Chen Weina , Rassidakis George , Routbort Mark , Jones Dan , Medeiros L. Jeffrey , Bueso-Ramos Carlos 
Published: October 2019

Abstract
Acute myelogeneous leukemia (AML) is a heterogeneous group of diseases. Mutations of the fms-like tyrosine kinase-3 (FLT3) gene of internal tandem duplication (ITD) type have been identified as one of the most frequent molecular defects in AML. However, even within FLT3 mutated/ITD positive cases, there is heterogeneity in clinical outcomes. In this study, we describe a biologically distinctive subgroup of AML in which blasts exhibit prominent nuclear invagination with frequent fishmouth-like nuclear indentation and had tendency of high frequency of FLT3/ITD. We identified 23 AML, FAB-M1 cases with fishmouth morphologic feature (study series, SS) diagnosed between 1998 and mid-2005 at our institution, which represents approximately 20% of AML, M1 cases and 1% of the entire AML cases. The age-matched control series (CS) of 20 AML, M1 cases without this nuclear feature were established to compare the incidence of FLT3 mutations, cytogenetics (CG), immunophenotype by multiparameter flow cytometry and clinical outcome. All the laboratory tests were performed on the bone marrow (BM) samples except one case using peripheral blood.
Characteristics of AML cases with and without fishmouth-like nuclear morphology age (y) sex M:F WBC x109/L Normal CG FLT3/ITD lack of CD34 lack of HLA-DR The values of age and WBC are the range and median. SS (n=23) 18–81 62 5:17 1.4–335, 48 15/22 (68%) 6/7 (86%) 16/23 (70%) 10/23 (43%) CS (n=20) 20–80 65 13:7 0.7–550, 35 9/20 (45%) 7/14 (50%) 2/20 (10%) 2/20 (10%) p value > 0.05 0.006 > 0.05 0.21 0.17 0.0004 0.017
As illustrated in the table, AML cases with fishmouth-like nuclear morphology are significantly associated with female gender and lack of expression of CD34 and HLA-DR. There is statistical trend toward high frequencies of normal cytogenetics (68% diploid by conventional karyotyping) and FLT3 mutation of ITD type (86%) in the study series. The white cell count, bone marrow blast percentage (82–98%, median 90% in SS vs. 76–97%, median 85% in CS) and expression of CD13, CD33 and CD117 are not significantly different. Electron microscopic examination demonstrated condensed collections of mitochondriae within the indented/invaginated nuclear pockets. Complete remission rate was higher in AML cases with fishmouth-like nuclear morphology (73% vs. 40%, p=0.032). However, there was no difference in the overall survival rate between these two groups (1-year survival of 38% in SS vs. 39% in CS). In conclusion, AML cases with fishmouth-like nuclear morphology may represent a rare but distinctive AML subgroup with specific clinical, immunophenotypic and genetic features. Further prospective studies of larger cohorts of patients are required to confirm the prognosis of this entity.
Study of Sequential Treatment of Newly Diagnosed De Novo AML Patients with DA and CAG Regimens as Remission Induction Therapy.
Authors: ping Sun Hui , Liu Wei Hong , Li Jun min , Chen Qiu sheng , Chen Yu 
Published: October 2019

Abstract
Objectives
To evaluate the efficacy and safety of sequential treatment of newly diagnosed de novo AML patients with DA and CAG regimens as induction therapy.
Methods
Those who were newly diagnosed as de novo AML (FAB classification criteria) were enrolled and DA regimen chemotherapy were administered. Bone marrow aspirates were performed and BM smears were examined at 48 hours since the end of chemotherapy. If severe hypocellularities were not achieved, the percentage of blasts in BM was between 20%–60% and peripheral WBC was in the range of (0.5–10) x109/L, the patients would receive CAG regimen therapy since 72 hours. Patients’ general status and the important parameters, such as peripheral blood count, liver function, renal function, thrombosis and hemostasis parameters were monitored throughout the course of the treatment and thereafter. When the clinical symptoms were relieved and peripheral blood counts returned to normal, or it was the end of the second or third week since the end of the CAG regimen, Bone marrow were examined again to evaluate the efficacy of the sequential therapy.
Results
14 patients consisted of 9 male and 5 female patients were enrolled. Out of them, 2 were M1, 5 M2, 4 M4 and 3 M5 according to FAB classification criteria. Median of blasts in BM were 38.5%(20%–60%) before CAG regimen. Of the 14 patients, 10 reached CR, 2 PR and 2 NR. CR rate was 71.4% (10/14) and total response rate was 85.7%(12/14). Time to achieve CR was on 15th(14th–29th)day medianly since the end of the treatment. During the CAG therapy courses, the nadir of peripheral blood cell counts and the time when it occurred were as follows: WBC 1.0(0.2–3.5)(x109/L),10(1–23)(d); Hb 57.5(44–69) (g/L), 10(1–27)(d)and PLT 11.5(10–65)(x109/L), 12(3–23)(d), respectively. Neutropenia (WBC<1.0x109/L) and thrombocytopenia (PLT<20.0x109/L) were lasted for 0(0–24) and 11(0–21)days, respectively. Median units of transfusions of platelets and red blood cells required by each patient were 3(0–10)(u) and 4(0–12)(u), respectively. The most commonly observed side effect of the regimen was bone marrow proliferation inhibition. Infections, usually respiratoy tract infections, were the second. However, sepsis was rare, which appeared in 1 out of 14 patients.
Conclusions DA and CAG regimens sequential treatment as remission induction chemotherapy in patients with newly diagnose de novo AML was highly effective and well tolerated. It would be beneficial for those who might not be sensitive enough to DA regimen chemotherapy only.
An Initial Analysis of an Allele-Specific Q-PCR Assay Which Detects Minimal Residual Disease in the Peripheral Blood of Follicular Lymphoma Patients.
Authors: Kautz Naomi , Ingola Diane , Denney Dan , Theriault Thomas 
Published: October 2019

Abstract
The monitoring of minimal residual disease (MRD) in peripheral blood cells of follicular non-Hodgkin’s lymphoma patients may provide important information regarding disease prognosis and response to therapy. The traditional method of assessing MRD in follicular lymphoma is by amplification of the t(14;18) translocation at the major breakpoint region (MBR/JH) locus. We are developing an allele-specific (ASO) quantitative PCR (Q-PCR) approach which has the ability to monitor tumor cells in the peripheral blood of patients who do not have a t(14;18) breakpoint within the MBR sequence. The patient-specific ASO assay amplifies immunoglobulin heavy chain sequence defined by unique CDR2/CDR3 (complementarity-determining region) primers. Both the generic MBR/JH and the patient-specific ASO Q-PCR assays were performed on peripheral blood lymphocytes collected before and after chemotherapy and following treatment with MyVax® personalized immunotherapy. MyVax® personalized immunotherapy (Id-KLH conjugate) is an active idiotype immunotherapy manufactured by Genitope Corporation that targets each patient’s unique tumor-associated immunoglobulin and is co-administered with GM-CSF. Preliminary Q-PCR data generated from these analyses could be grouped into MBR positive and MBR negative sets. Initial results have shown MBR values can track ASO values at sequential time points during therapy. In the MBR negative group, we were able to monitor tumor-specific cells present in peripheral blood by the ASO assay. In terms of MRD assessment in follicular lymphoma patients, the allele-specific Q-PCR assay may provide a valuable extension of analysis beyond the standard approach of solely monitoring MBR/JH values. Genitope Corporation is focused on the development of novel means for both treating follicular lymphoma and monitoring the patient’s MRD in a patient-specific manner.
Isophosphoramide Mustard-Lysine (ZIO-201): Active Moiety of Ifosfamide (IFOS) without Kidney Toxicity.
Authors: Taub Mary , Springate James E. , Zaki Emed , Struck Robert 
Published: October 2019

Abstract
Ifosfamide (IFOS) and cyclophosphamide (CPA) are widely used anti-cancer drugs. Both are pro-drugs which must be metabolized before their metabolites cross-link DNA. Use of these drugs, especially high-dose IFOS, is complicated by substantial adverse effects of metabolites not directly involved in DNA-cross-linking including kidney, bladder and CNS toxicities. Recently, a lysine-stabilized form of isophosphoramide mustard (IPM-lysine; ZIO-201), an IFOS-metabolite, was developed. ZIO-201 directly cross-links DNA and is active against human cancer cell lines and in mice with human cancer xenografts. To evaluate nephrotoxicity of ZIO-201 we used an in vitro primary rabbit kidney proximal tubule (RPT) cell culture system, which retains many of the characteristics of renal proximal tubule cells including a polarized morphology, a Na+/glucose cotransport system and a p-aminohippurate transport system, glutathione status, and hormone responses (including a parathyroid hormone sensitive adenylate cyclase). We studied effects of ZIO-201 and other IFOS metabolites on primary RPT cells. IFOS is activated by the cytochrome P450 system producing a mixture of 2- and 3-dechloroethylifosfamide and resulting in formation of chloroacetaldehyde (CAA). Also, metabolism of IFOS by the cytochrome P450 system produces 4-hydroxyifosfamide, ultimately resulting in the production of acrolein (ACR). CAA and ACR are IFOS metabolites implicated in kidney toxicity in humans. Effects of ZIO-201 and CAA on the viability of confluent monolayers of primary rabbit kidney proximal tubule (RPT) cells were tested at 15–100 μM. Viability was determined by neutral red dye uptake by the primary cultures after 8 h incubation. ZIO-201 did not significantly reduce viability of primary RPT cells; the LD50 for CAA was about 40 μM with complete inhibition at 75 μM. Effects of 4-hydroperoxyifosfamide (HIFA), a synthetic prodrug of 4-hydroxyifosfamide and ACR were also studied. Although ACR had an LD50 of 80 μM HIFA did not significantly affect viability. The IFOS metabolite 4-hydroxyifosfamide, was prepared from 4-hydroperoxyifosfamide (1). At 200 μM, 4-hydroxyifosfamide resulted in extensive cell death after 4 h incubation. These data show that ZIO-201 avoids the kidney toxicity caused by metabolites of pro-drugs like IFOS. Whereas 2 IFOS metabolites, CAA and ACR were cytotoxic to kidney cells, ZIO-201 was not. An explanation for the toxic effects of 200 μM 4-hydroxyifosfamide t might be metabolism to ACR in vitro.
Disruption of Leukemia/Stroma Cell Interactions by CXCR4 Antagonist AMD3465 Enhances Chemotherapy-Induced Apoptosis in AML.
Authors: Zeng Zhihong , Konopleva Marina , Nowak Billie J. , Plunkett William , Borthakur Gautam , Estey Elihu , Champlin Richard , Bridger Gary , Andreeff Michael 
Published: October 2019

Abstract
Chemokine receptor CXCR4 is critically involved in the migration of hematopoietic cells to the stromal derived factor (SDF-1α)-producing bone marrow microenvironment. We and others have previously demonstrated that stroma/leukemia interactions mediate protection of leukemic cells from chemotherapy-induced apoptosis (Konopleva, Leukemia2002:1713; Burger Blood2000: 2655). Using AMD3465, the second-generation small-molecule CXCR4 inhibitor with a greater potency than AMD3100, we tested the hypothesis that CXCR4 inhibition interferes with stromal/leukemia cell interactions resulting in increased sensitivity to chemotherapy. Our results showed that AMD3465 inhibited surface expression of CXCR4 on AML cell lines in a dose dependent manner. AMD3465 (1μM) significantly inhibited SDF-1α and stromal (MS-5)-induced migration of OCI-AML2 cells (78% and 54% inhibition, respectively), U937 cells (71% and 41.3%) and diminished SDF-1α- or stromal-induced migration of leukemic blasts from four primary AML samples tested (SDF-1α, 43.4 ± 8.6%, MS-5, 38.4 ± 8.5% inhibition). In in vitro co-culture systems, stromal cells significantly protected leukemic cell lines and primary AML cells from spontaneous and chemotherapy induced apoptosis (p<0.01; p<0.001). Measurements of intracellular Ara-CTP levels determined by HPLC showed that stromal cells diminished incorporation of Ara-C into leukemic cells by 20%. AMD3465 enhanced AraC- and Busulfan-induced apoptosis by 44% and 69%, respectively. Western blot revealed that AMD3465 downregulated AKT signaling in AML cells. Most importantly, it decreased stroma-mediated protection from AraC-induced apoptosis in five out of ten primary AML samples with surface expression of functional CXCR4 (mean increase, 29.9±19.5% compared to chemotherapy alone). Curiously, the highest sensitization was observed in a sample from AML patient harboring Flt3/ITD mutation (Ara-C, 30.3% annexinV(+); Ara-C+AMD, 62.8%), confirming recently documented role for Flt3/ITD in modulation of CXCR4 signaling (Fukuda, Blood2005:3117). Taken together, our data suggest that SDF-1α/ CXCR4 interactions contribute to the resistance of leukemic cells to chemotherapy-induced apoptosis. Disruption of these interactions by the potent CXCR4 inhibitor AMD3465 represents a novel strategy for targeting leukemia cell/bone marrow microenvironment interactions. A clinical trial testing this concept in patients with AML is in preparation.
Novel Combination of Gallium Nitrate, Rituximab and Dexamethasone for the Treatment of Relapsed or Refractory Diffuse Large B-Cell Lymphoma, Mantle Cell Lymphoma or Transformed Follicular Lymphoma.
Authors: Smith Scott E. , Stiff Patrick J. , Rodriguez Tulio , Toor Amir , Klein Jared 
Published: October 2019

More than 56,000 new cases of non-Hodgkin’s lymphoma (NHL) will be diagnosed this year in the United States. Despite advances in treatment modalities such as radiation, biologic agents, and cytotoxic chemotherapeutic regimens, only 25–30% of these patients will be cured of their disease. Standard salvage regimens such as DHAP, ESHAP, ICE and EPOCH have proven efficacy at the cost of increasing toxicity and hospitalization costs. The reported response rates for these are on the order of 50–75% as first-line salvage regimens. However, as our aging patient population develops worsening performance status and co-morbidities, it seems appropriate to develop effective lymphoma treatments, which have fewer toxicities and lower costs for administration. One approach is to combine non-myelosuppressive therapies. One non-myelosuppressive agent, which has efficacy in lymphoma, is gallium nitrate. Investigation of gallium nitrate for cancer treatment dates back to the 1970’s. While it is currently approved for the treatment of bisphosphonate resistant hypercalcemia of malignancy, it has also been shown to inhibit ribonucleotide reductase and bind transferrin and potentially complex with the transferrin receptor, which is highly expressed in intermediate and aggressive histology lymphomas. It appears that the binding of the transferrin receptor on the lymphocyte as well as its inhibition of ribonucleotide reductase, eventually impairs iron metabolism, which is a necessary component of the intracellular cytochrome systems/mitochondrial function and ultimately oxidative phosphorylation.
The current study is a phase II clinical trial investigating the combination of gallium nitrate, rituximab and dexamethasone (GaRD) for relapsed or refractory DLBCL, MCL or transformed follicular lymphomas. The gallium nitrate is given at 200mg/m2 CIV days 1–7, rituximab 375mg/m2 IVPB day 1 and dexamethasone 40 mg po days 1–4. Eligible patients must have proven relapsed or refractory disease and have a SWOG PS <3. Patients may have failed prior ASCT or allogeneic SCT. The accrual goal is 37 patients. We have enrolled 14 patients on study to date and have 12 evaluable patients; as part of the ongoing safety evaluation for this study, we have the following results: ORR 9/12 (75%); CR 2/12 (17%); PR 7/12 (58%); SD 2/12 (17%); and PD 1/12 (8%). Most of these patients were refractory to prior salvage regimens 8/12 (67%), including ESHAP, DHAP or high-dose cyclophosphamide. No patients developed grade 3 or 4 toxicities, with the exception of grade 4 lymphopenia and grade 3 anemia (most likely due to transferrin receptor binding).
Conclusions: gallium nitrate, rituximab and dexamethasone (GaRD) appears to be an effective and relatively non-toxic salvage regimen for patients with relapsed DLBCL, MCL or transformed FL.