bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cells Lacking the PolQ Polymerase Are Moderately Sensitive to Ionizing Radiation and the Oxidant Induced Toxicity of Paraquat and Bleomycin.
Authors: Goff Julie P. , Epperly Michael W. , Shields Donna S. , Smith Tracy , Seki Mineaki , Wang Hong , Wittschieben John P. , Wood Richard D. , Dertinger Stephen D. , Torous Dorothea K. , Greenberger Joel S. 
Published: October 2019

Abstract
The DNA polymerase POLQ (pol theta) can bypass abasic sites in DNA and thymine glycol, a common product of reactive oxygen species damage to DNA. POLQ may also have a back-up role in base excision repair. To explore the role of POLQ in tolerance of DNA damage following irradiation, and exposure to agents that induce oxidative stress we established bone marrow stromal cell lines from the PolQ+/+ and PolQ−/− mice. Irradiation survival curves were obtained for PolQ +/+, and 2 clones of PolQ −/− cells. Dose-response experiments using the free radical generators paraquat, hydrogen peroxide and bleomycin were done on 2 clones each of PolQ +/+ and PolQ −/− cells. There was no significant difference over 23 weeks in cumulative granulopoiesis in LTBMCs from PolQ +/+ and PolQ −/− mice. PolQ+/+ cells and 2 independent clones of PolQ−/− cells were exposed to 0 – 8 Gy of gamma radiation, and colony forming ability was measured. PolQ −/− cells were more sensitive to irradiation as shown by decreased Do from 1.38 + 0.06 Gy for PolQ +/+ cells compared to 1.27 + 0.16 and 0.98 + 0.10 (p = 0.0316) Gy for PolQ −/− clones 1 and 3 respectively. Micronucleated reticulocytes were measured in peripheral blood of PolQ+/+, +/− or −/− mice after irradiation with 75 cGy or 700 cGy. Before irradiation, PolQ−/− mice had a higher fraction of MN- reticulocytes compared to PolQ+/+. Forty hr after irradiation, MN reticulocytes in PolQ−/− mice increased to 8.53 ± 1.48%, compared to 2.35 ± 0.12% and 2.55 ± 0.17% in PolQ+/+ mice (p = 0.0032). PolQ +/+ and PolQ −/− cells were exposed to paraquat (0–20 mM), hydrogen peroxide (0–40 uM) and bleomycin (0–5 ug/ml) for 1 hr prior to plating. After 7 days, the cells were stained and colonies of 50 cells or greater were counted. Statistical analysis was done using ANCOVA. PolQ −/− cells were modestly more sensitive to paraquat (p< 0.0001) and bleomycin (p< 0.0001) than PolQ +/+ cells. When the cells were treated with hydrogen peroxide, there was no significant difference in surviving fraction (p=0.7327) between the PolQ +/+ and the −/− cells. The moderate but significant sensitivity of POLQ-defective cells to bleomycin, ionizing radiation and paraquat supports the hypothesis that POLQ normally participates in helping cells tolerate DNA damage. The increased frequencies of spontaneous and radiation-induced micronuclei in peripheral blood red cells indicate that POLQ has a role in maintaining genomic integrity via limiting double-strand DNA breaks.
Use of Single Course IV Fludarabine in Treatment of Hypocellular Bone Marrow (Aplastic Anaemia and MDS) Patients.
Authors: Apte Shashikant J. , Kelkar Anjali J. , Melinkeri Sameer , Melinkeri Varsha , Vaidya Sujata , Sayaji Bhavana 
Published: October 2019

Abstract
Young patients with Hypocellular marrow (SAA, VSAA, MDS) ideally need ALLO SCT or Triple Immunosupression with ATG + Cyclosporin + Steroids therapy. However these therapies are very expensive and affordable to very few in third world poor countries. Majority of these patients are treated with androgens with dismal outcome. IV Fludarabine has been used in the setting of ALLO SCT in the preparative regimens and the immunosupressive effect of IV Fludarabine is well established.We decided to assess effect of IV Fludarabine for these patients. The comparative cost is as follows in our country: Allo SCT-$25000/-, ATG+ CYA Therapy- $ 14000/-, IV Fludarabine150mg / m2 - $ 1300/-. At our center we approximately diagnose 75 new cases of SAA, VSAA every year. About 10% undergo ALLOSCT, 15% are treated with ATG+CYA. Approximately 75% patients are treated with androgens. We started this study in patients who could not be treated upfront with standard therapy. W e totally treated 55 patients with SAA, VSAA upfront with IV FLUDARABINE 30mg/m2/day for five successive days. 20 / 55 (36%) patients showed good response and became RBC, Platelet transfusion independant.All these patients achieved ANC > 1500/cmm. 21 / 55 (38%) patients died in first six months. 14 /55 (25%) patients showed no response. Thus SINGLE COURSE IV FLUDARABINE 150 mg /m2 can be a good, cheaper treatment option for newly diagnosed SAA, VSAA.
Patient data Study Period Jan 2001 June 2007 DOSE: IV Fludarabine 30mg / m2 x 5 days Total Patients 55 Age Group 15 – 56 years Males - 37 females -18 Follow up 2 – 54 months Median follow up 36months Haemogram ranges at diagnosis Hb: 3 - 7.4 gm% ANC : 100 – 600cmm Platelet count - 3 -12 x 10′9/l Dysplasia - 12 /55 Cytogenetics 4/55 patients had cytogenetic abnormality : two had del 7 two had del7q 21/55 patients had normal cytogenetics 30/55 patients no metaphases seen on cytogenetics All cellular transfusions were irradiated with 2500 rads. Response 20 / 55 -Response 14 / 55 - No Response 21/55 - Deaths
Expansion of Umbilical Cord Blood Hematopoietic Stem Cells for Clinical Use.
Authors: Peters Rowayda , Hutter Gregor , Aguzzi Adriano , Heikenwaelder Mathias , Renner Christoph , Knuth Alexander 
Published: October 2019

Abstract
Umbilical cord blood (UCB) is an alternative donor source for allogeneic hematopoietic stem cell transplantation. However, transplantation in adults is frequently limited by the small number of cells available in a unit. We have previously developed the technology to expand hematopoietic stem cells (HSC) and stromal/mesenchymal stem cells (SMSC) from all UCB frozen samples (13, 3). The incubation of thawed UCB mononuclear cells (MNC) in the presence of SCF (25ng/ml), FLt-3 (25ng/ml), MGDF (10ng/ml) & IL-6 (20ng/ml) and 10% human serum in stroma-free liquid culture not only generated long term expansion of transplantable UCB HSC (non-adherent). Also, long-term expansion of SMSC (adherent cells) was successful. In order to upscale the expansion to use for clinical applications, we analyzed 3 frozen UCB comparing the expansion from 24-well plates (previous) versus expansion in bags (VueLife™) after 10 and 14 days of culture. Results show substantial expansion in total cell count (TCC, 4.8, 10.9: 2.4, 3.8 fold) at d10 and d14 from wells & bags respectively. TCC increased further in the presence of SMSC (38% & 33% in CD34+ cell count cultured in wells). CD133+CD34+CD38- HSC multiplied (11, 25 fold, d10/d14 & colony forming cells (CFC) 19 fold at d14 bags respectively). Heterogeneous cell populations were detected after d 14 in bags: T and B -lymphoid (%CD3/%CD19; 65/4:50w/3), megakaryocytic (%CD61; 7:4) and myeloid (%CD33; 31:43) at d0/d14 respectively. Further, expanded cells (250,000) containing a small number of CD133+CD34+CD38- (15,000–30,000) were injected into the liver of sub-lethally irradiated newborn Balb/C Rag2-Cγ−/− mice. Our preliminary data show no engraftment from cells expanded in wells and bags after 6 weeks of transplantation from d10 cultures (human CD45 + <1%). However, positive engraftment in mouse PB was detected from cells expanded for 14 d (wells, 1.16–2.5% & bags, 1.21–3.9%) as compared to control mice (CD45; 30% PB & 70% BM) receiving selected CD34 + (300,000 CD34+ at d0). Primitive repopulating cells (PRC), and multilineage human CFC were detected after transplantation. On d14 of HSC expansion, UCB MNC derived adherent cells (SMSC) were enriched by trypsinization. SMSC were established in serum free and serum plus culture as well. The immunophenotype of harvested SMSC was CD29+, CD44+ and CD45−, CD34− and CD133− at percentages + >90%. Following repeated trypsinization, SMSC count increased 41–96 folds. CFU-Fibroblast colonies (92–173) were generated from 104 SMSC after 2 weeks in MesenCult medium. We have previously differentiated SMSC into hepatocytes. Now we also generated adipocytes in an induction medium containing, Insulin, dexamethasone & indomethacin. SMSC formed Oil-droplet vacuoles in the cytoplasm in 3 weeks. The culture conditions we defined to maintain UCB PRC should be developed clinically. SMSC described herein exhibit in vitro properties of multipotent stem cells.
Reduction of Prothrombin in Mice Carrying a Conditional Knockout Allele Attenuates the Development of Paralysis Associated with Experimental Encephalitis.
Authors: Mullins Eric S. , Kombrinck Keith W. , Flick Matthew J. , Shaw Maureen A. , Sun William , Degen Sandra J. , Akassoglou Katerina , Degen Jay L. 
Published: October 2019

Abstract
Multiple sclerosis (MS) is common and often devastating chronic inflammatory disease leading to demyelination in the central nervous system and relapsing neurological deficits, including paralysis and vision loss. MS lesions are characterized by blood-brain barrier disruption leading to perivascular deposition of fibrin that correlates with areas of microglia activation and myelin damage. Consistent with multiple studies showing that hemostatic factors can serve as important modulators of the inflammatory response in vivo, we recently identified fibrin(ogen) as a novel regulator of microglial activation/differentiation and showed that fibrinogen plays a causative role in the development of inflammatory demyelination in experimental autoimmune encephalomyelitis (EAE), an established animal model for MS. A working hypothesis that has emerged is that Mac1-mediated microglial cell engagement of fibrin-rich matrices deposited within MS plaques drives activation events leading to neuronal destruction. To better define contribution of the thrombin/fibrin axis in demyelinating neuroinflammatory disease, we challenged conditional “floxed” fII knockout mice carrying low levels (∼10% of normal) of circulation prothrombin (fII) with EAE initiated by immunization with myelin oligodendrocyte peptide. Wild-type mice challenged with EAE typically began to develop overt neurological symptoms within two weeks of immunization. Clinical disease progressed from simple loss of tail tone to ataxia and, ultimately, fore- and hind-limb paralysis. When cohorts were clinically scored in a fashion blinded to animal genotype, prothrombin-deficient mice were found to exhibit significantly reduced disease severity than the control mice tracked in parallel. The timing of disease onset was similar in fII-deficient mice as in controls, consistent with the hypothesis that fII-deficiency diminishes microglial activity. In contrast to control mice that uniformly developed clinically apparent disease, the two-thirds of fII-deficient mice develop either no disease or very mild disease typically limited to the simple loss of tail tone. More detailed studies of fII-deficient mice, including comparative studies of CNS histopathology, are now underway to more fully define the benefits and liabilities of diminished prothrombin on CNS disease. Complementary studies are also underway with gene-targeted mice expressing a mutant form of prothrombin (fII with “specificity switch” favoring protein C over procoagulant substrates). Both prothrombin and fibrinogen are powerful determinants of inflammatory CNS demyelinating disease and a more detailed understanding of the contribution of these factors to disease progression may reveal novel therapeutic strategies for the attenuation of this and other neuroinflammatory diseases.
Relation between FLT3 Mutations, High Flow Cytometric FLT3 Expression and CXCR4 Expression in CD34+ Cells in Adult Acute Myeloid Leukemia.
Authors: Yang Li , Wu Xiao Jin , Wu De Pei 
Published: October 2019

Abstract
Objective
To evaluate the prevalence of FLT3 mutation-interna tanden duplication in adult acute leukemia patients and its clinical significance To evaluate the surface expression of FLT3 difference between AML and non malignant hemopoietic disease, and the possibility of detection of CD135 being included in the diagnostic immunophenotyping of acute leukaemias To study if constitutive FLT3 activity induce cell surface expression of CXCR4 in CD34 cells .
Methods
Genomic DNAs from 95 cases of AML and 22 cases of ALL were screened bypolymerase chain reaction and gel electrophoresis for FLT3-ITD mutations. Flow cytometry is used to investigate FLT3 expression in 68 cases AML and 8cases non-malignant hemopoietic disease To investigate FLT3 expression using flow cytometry between 56 cases FLT3-ITD- AML and 12 cases FLT3-ITD+ AML.
Results
FLT3-ITD were found in 15 (16%) of 95 AL patients,including 3/ 15 of M1, 4/ 24 of M2, 2/ 18 of M3, 5/ 29 of M5, 1/4 of M6.Significantly more FLT3 mututations were found in AML M1, M5. No FLT3-ITD was found in 22 ALL. FLT3-ITD was associated with a higher peripheral white cell count (P<0.05) and a lower complete remission rate (P<0.05).and was more prevalent in patients with normal karyotype(P<0.05) an t(15;17). High level expression of CD135 in acute myeloid leukaemias has been demonstrated by flow cytometry .Dection of CD135 maybe included in the diagnostic immunophenotyping of AML. Significant difference of FLT3 expression between FLT3 ITD posive cases and FLT3 ITD negative cases.
Eltrombopag Decreases Proliferation of Human Leukemia and Lymphoma Cell Lines In Vitro.
Authors: Erickson-Miller Connie L. , Payne Parrish L. , Moore Sherri , Wert Sherryann , May Richard D. 
Published: October 2019

Abstract
Eltrombopag is a novel, oral platelet growth factor that interacts with the thrombopoietin receptor on bone marrow progenitors to stimulate megakaryocyte and platelet production. Clinical trials have demonstrated that eltrombopag increases platelet counts in healthy volunteers and in patients with idiopathic thrombocytopenic purpura (ITP) and hepatitis C. Because thrombopoietin (TPO) receptors are expressed on lymphoma and leukemia cell lines and recombinant human TPO (rhTPO) has been shown to stimulate their proliferation in vitro, experiments were conducted to evaluate the effects of eltrombopag on the proliferation of leukemia and lymphoma cells in vitro. Five leukemia and lymphoma cell lines (ie, CCRF-CEM, K562, MOLT-4, RPMI-8226 and SR) representing lymphoblastic T cell leukemia, chronic myelogenous leukemia, acute lymphoblastic T cell leukemia, plasmacytoma and immunoblastic large cell lymphoma, were exposed to eltrombopag (0.1–40 ug/mL), rhTPO (100 ng/mL) or both for 3 days. Proliferation was measured by 18-hour incubation with tritiated thymidine incorporation. As expected, rhTPO alone led to a small, statistically significant increase in the proliferation of CCRF-CEM and RPMI-8226 cells, while it had no effect on the other cell lines. However, the addition of eltrombopag to rhTPO negated the increased proliferation seen with rhTPO alone. Even more remarkable, treatment with eltrombopag alone inhibited the proliferation of all five leukemia and lymphoma cell lines with an IC50 range of 0.56 to 5.9 ug/mL and 100% inhibition (IC100) of thymidine incorporation at 10 ug/mL (Table). Previous studies have shown that eltrombopag induces proliferation and differentiation of bone marrow progenitor cells in vitro and increases platelet counts in vivo. While eltrombopag and rhTPO interact with the TPO receptor to stimulate the production of platelets, there are differences in the site of receptor interaction and their signaling pathways. The findings of the current study suggest that eltrombopag may inhibit the proliferation of leukemia and lymphoma cell lines unlike the effect that has been demonstrated with rhTPO. Interestingly, eltrombopag may also serve to mitigate rhTPO-mediated proliferation of malignant hematologic cell lines. These findings merit further evaluation of the effects of eltrombopag on leukemia, lymphoma, and solid tumor cell proliferation.
Cell line IC50 (ug/mL) IC100 (ug/mL) CCRF-CEM 0.74 10 K562 1.80 10 MOLT-4 0.56 4 RPMI-8226 5.90 10 SR 0.77 4
Anti-Glycophorin C Induces a Loss of Mitochondrial Membrane Potential but Fails To Activate Apoptotic Caspases.
Authors: Denomme Gregory A. , Micieli Jonathan , Shu Jenny , Wang Dan , Fernandes Bernard J. 
Published: October 2019

Abstract
The human erythrocyte transmembrane sialoglycoprotein, glycophorin C (GYPC), plays a functional role in regulating red cell shape and mechanical stability. Antibodies to GYPC cause hemolytic disease of the fetus and newborn (HDFN) that is associated with classical Fcγ receptor-mediated phagocytosis. However, in vitro clonogenic studies with cord blood progenitor cells suggest that anti-GYPC also suppresses erythropoiesis, which is consistent with the observations of severe and early fetal anemia and late onset neonatal anemia [Transfus Med2005;15:125–32]. The mechanism of the suppressive effect on erythropoiesis is unknown. The K562 erythroleukemic cell line treated with anti-GYPC is a potential model system to study the suppressive effect of anti-GYPC. The present in vitro studies were designed to confirm the effect of anti-GYPC on K562 cell growth and viability, and to evaluate changes in mitochondrial membrane potential, phosphatidylserine (PS) expression, propidium iodide (PI) binding, and caspase activation. K562 cells fail to grow in the presence of anti-GYPC confirming earlier CFU-E/BFU-E studies [Brit J Haematol2006;133:443–4], and increased the exofacial expression of PS/PI over time. This process was caspase-independent as demonstrated by the failure of Z-VAD, a caspase inhibitor, to reverse growth inhibition and PS/PI expression. A loss of mitochondrial membrane potential was demonstrated using JC-1, a cationic dye that is sensitive to potential-dependent accumulation or loss in mitochondria. There was a 50% increase in K562 cell mitochondrial membrane potential disruption after 2 days of culture with anti-GYPC (see figure). Morphological examination of May Grunwalde Giemsa-stained K562 cells treated with anti-GYPC for 2 days showed a decrease in mitotic activity compared to isotype treated cells. By day 4, the anti-GYPC treated cells were showing evidence of plasma membrane damage and cell death resulting from fragmentation and dissolution of the cytoplasm. The addition of hemin, an oxidative form of iron protoporphyrin IX known to induce erythroid differentiation of K562 cells, to anti-GYPC treated cells reversed growth inhibition by 45% but did not prevent the loss of mitochondrial membrane potential. Overall, although caspases appear to be unimportant in anti-GYPC induced cell death, the mitchondria play an important role as the early events leading to antibody-mediated suppression of erythropoiesis.
Mitochondrial Membrane Potential Disruption by Anti-GYPC Mitochondrial Membrane Potential Disruption by Anti-GYPC
The Prolonged Time to Progression with Pegylated Liposomal Doxorubicin + Bortezomib Versus Bortezomib Alone in Relapsed or Refractory Multiple Myeloma Is Unaffected by Extent of Prior Therapy or Previous Anthracycline Exposure.
Authors: Blade Joan , Miguel Jesus San , Nagler Arnon , Sonneveld Pieter , Spencer Andrew , Robak Tadeusz , Hennessey Kathleen , Mundle Suneel D. , Zhuang Sen H. , Harousseau Jean-Luc , Orlowski Robert Z. 
Published: October 2019

Abstract
Recently, a significant improvement in time to progression (TTP) was reported for pegylated liposomal doxorubicin (PLD) + bortezomib combination therapy vs. bortezomib monotherapy in a phase III randomized trial in relapsed or refractory multiple myeloma (RRMM) (Orlowski, JCO 2007). This pre-specified analysis assessed the efficacy of PLD+bortezomib in RRMM based on the number of prior lines of therapy and previous anthracycline exposure status. Patients with ≥1 prior therapy were randomized to receive PLD at 30 mg/m2 on day 4 and bortezomib at 1.3 mg/m2 on days 1, 4, 8, and 11, or bortezomib alone for up to eight 21-day cycles, or at least 2 cycles beyond CR, or optimal response unless disease progression, or unacceptable toxicities occurred. The baseline beta-2 microglobulin levels were comparable between the subgroup with ≥2 prior treatments and that with 1 treatment (median 3.98 vs. 4.01 mg/L), as well as between anthracycline-exposed and -naïve patients (median 3.91 vs. 4.55 mg/L). The improved TTP reported previously with PLD+bortezomib over bortezomib in the total study population was similarly observed in all four subgroups of patients (patients with ≥2 lines of prior therapy or 1 prior therapy, anthracycline-exposed (median 144 mg/m2 in both treatment arms) or -naïve groups (Table)), indicating a consistent therapeutic benefit favoring the PLD+bortezomib combination. Furthermore, TTP was comparable for the PLD+bortezomib combination between groups with ≥2 lines of prior therapy vs. 1 prior therapy (heterogeneity test, p=0.523), as well as patients who were anthracycline-exposed or -naïve (heterogeneity test, p=0.716). Incidence of treatment-related SAEs, grade 3/4 neuropathy, and symptomatic cardiac events was comparable between treatment arms in each subgroup (Table). PLD-related hand-foot syndrome was also <10% in all PLD+bortezomib subgroups. These results demonstrate that PLD+bortezomib combination therapy significantly improves TTP compared to bortezomib alone, regardless of the number of prior lines of therapy, or anthracycline exposure.
Prolonged Overall Survival with Lenalidomide Plus Dexamethasone Compared with Dexamethasone Alone in Patients with Relapsed or Refractory Multiple Myeloma.
Authors: Weber Donna , Knight Robert , Chen Christine , Spencer Andrew , Yu Zhinuan , Zeldis Jerome , Olesnyckyj Marta , Dimopoulos Meletios 
Published: October 2019

Abstract
Introduction: Lenalidomide (Len), an analog of thalidomide (Thal) is a novel, oral, immunomodulatory agent that is effective against multiple myeloma (MM). In 2 prospective, randomized, double-blind, placebo-controlled phase III trials, Len with dexamethasone (Dex) induced a significantly higher overall response (OR) rate and complete remission (CR) rate, as well as longer time-to-progression (TTP) in comparison with Dex alone. Here, we investigate the long-term overall survival (OS) with Len/Dex.
Methods: We evaluated the pooled results from both randomized trials (MM-009, MM-010) of 704 patients who had relapsed or refractory MM, without prior resistance to Dex, who received either Len (25 mg daily for 3 weeks every 4 weeks), or placebo. Dex was given at 40 mg on days 1–4, 9–12, 17–20 every 4 weeks for 4 cycles. From cycle 5 onwards, Dex was given at 40 mg on days 1–4 only. Response rate and TTP are based on data obtained before unblinding (June 2005 [MM-009] and August 2005 [MM-010]). Follow-up data on OS were obtained up to January 2007. Forty-seven percent of patients who received placebo/Dex crossed over to receive Len +/− Dex.
Results: Of 704 patients, 353 were treated with Len/Dex and 351 with Dex alone. Baseline characteristics were well balanced between patients receiving Len/Dex and those receiving Dex alone. Median TTP, OR, and CR were significantly improved in patients treated with Len/Dex compared with Dex alone (Table). Of patients who progressed on Dex alone prior to unblinding, or were found to be receiving Dex alone after unblinding, 47% crossed over to Len +/− Dex. Despite these patients crossing over to Len +/− Dex at progression or at the time of unblinding, the OS was significantly improved in patients treated with Len/Dex compared with Dex alone (hazard ratio 1.295; 95% confidence interval 1.040–1.614; p=0.02). Median OS in the Len/Dex group was 35 months and 31 in the Dex alone group (p<0.05). Median OS was also significantly longer with Len/Dex compared with Dex alone in patients with more than 1 prior therapy (32.4 months versus 27.3 months, p<0.05). Similar median OS was observed with Len/Dex and Dex alone in patients with 1 prior therapy (median OS not yet reached and 35.3 months, p=0.24).
Conclusion: With increased follow-up and despite cross-over, patients treated first with Len/Dex had significantly improved OS compared with those treated with Dex alone.
Len/Dex (n=353) Dex alone (n=351) P value OR, % 60.6 21.9 <0.001 CR, % 15.0 2.0 <0.001 Median TTP, months 11.2 4.7 <0.001 Median OS, months 35.0 31.0 <0.05 Median OS in patients with 1 prior treatment, months not yet reached 35.3 0.24 Median OS in patients with >1 prior treatment, months 32.4 27.3 <0.05