bluuebunny/crossref_metadata_2025_split huggingface dataset:
Alteration in Melphalan Conditioning Regimen for Multiple Myeloma: Reflection on Therapeutic Benefit and Length of Hospital Stay.
Authors: Gopaluni Srivalli , See Heidi , Coyle Thomas , Gentile Teresa C. 
Published: October 2019

Abstract
BACKGROUND: Melphalan is the most common conditioning regimen prior to autologous stem cell transplant in patients with Multiple Myeloma. Conventionally Melphalan was administered at a dose of 200mg/m2 for 2 days[100 mg/m2 each day] prior to transplantation on days -4 and -3 followed by autologous stem cell infusion on day 0. Recently we changed the scheduling of Melphalan to 200mg/m2 given on day-2 and stem cell infusion on day 0.
OBJECTIVE: A retrospective analysis was done to assess the effect of this change in Melphalan regimen on length of stay in hospital and gastrointestinal toxicity.
METHODS: We performed a retrospective analysis of 20 patients receiving melphalan as conditioning regimen prior to autologous stem cell transplant. All patients received similar induction therapy and were transplanted as part of their initial therapy. Assessment including maximum oral score per day of transplant (using the oral assessment guide developed by the University of Nebraska Medical Center) and number of days with nausea/vomiting/diarrhea were analyzed to compare gastrointestinal toxicity.
PATIENTS: Records on twenty patients with multiple myeloma undergoing autologous stem cell transplant from January 2003 until August 2006 were reviewed. Study population included10 females and 10 males. Average age was 60 yrs in melphalan 200mg/m2 over 2 days group and 55 in melphalan 200mg/m2 over one day group.
RESULTS: The average length of hospital stay was 17.4 days in the Melphalan 200mg/m2 over 2 days group and 15.4 days in the Melphalan 200 mg/m2 over 1 day group. The average number of days of reported nausea/vomiting and diarrhea was 7.1 in the Mel 200 mg/m2 over1 day and 7.6 days in the Mel 200mg/m2 over 2 day group. The average peak daily oral score in the Mel 200mg/m2 over 2 days was slightly higher (14 vs 12) and the duration of reported abnormal oral scores was slightly longer for the patients in the Mel 200mg/m2 over 1 day group (13 days vs. 10 days).
CONCLUSION: The major advantage of delivering Melphalan over one day as opposed to two days is decreased length of hospital stay and thereby the costs. However this does not appear to be at the expense of increased gastrointestinal toxicity. Although the duration of mucositis appears to be longer in the Melphalan 200mg/m2-1 day group, this did not lead to a prolonged length of stay in our patients. Reported nausea/vomiting/diarrhea was not significantly different in both groups.
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The Effects of Rapamune on NK Cell Post Bone Marrow Transplantation in Mice.
Authors: Chen Guanghua , Wu De Pei , Yang Ming Zhen , Tang Xiao Wen , Sun Ai-ning 
Published: October 2019

Abstract
Natural killer(NK) cells are innate effector cells of the immune system, believed to limit viremia and tumor burden before the onset of adaptive T and B cell immunity. NK cells are potent effector cells in allogeneic bone marrow transplantation. NK cell activity is partially controlled through interactions between killer Ig-like receptors on NK cells and their respective HLA class I ligands. Immunosuppressive agents including cyclosporin, FK506 and Rapamune are utilized in clinical hematopoietic stem cell transplantation. Little is known about the effects of immunosuppressive agents on NK cells post bone marrow transplantation. The in vivo effects of Rapamune on NK cells were determined in an allogeneic bone marrow transplantation model. Splenic NK cell levels in recipients treated with Rapamune decrease markedly. NK cell proliferation and function are significantly decreased in the presence of Rapamune. Studying the differential effects of immunosuppressive drugs on NK cell function is critical in clinical hematopoietic stem cell transplantation.
Uncovering the Epigenetic Pathomechanism in 13q14.
Authors: Mertens Daniel , Ruppel Melanie , Philippen Angela , Fleig Verena , Brakel Bianca , Stilgenbauer Stephan , Lichter Peter , Doehner Hartmut 
Published: October 2019

Abstract
INTRODUCTION: Deletions in chromosomal band 13q14.3 distal to RB1 occur in a variety of human neoplasms like B-cell chronic lymphocytic leukaemia (CLL), indicating a tumor suppressor mechanism in this region. Intriguingly, several characteristics of the region of interest point to an epigenetic pathomechanism:
candidate genes lack point mutations, yet these genes are downregulated in tumors, the presence of large non-coding RNA genes in 13q14.3 is reminiscent of imprinted regions where only one gene copy is active.
The data we show here led us to propose a novel oncogenic mechanism where already in healthy tissue only one gene copy is active while one gene copy is randomly chosen for silencing. Loss of the single active copy is then sufficient for complete loss of gene function in tumor cells. Currently we are trying to identify the (epi-)genetic element that controls the whole locus.
AIM: Identification of the epigenetic regulatory mechanism localized in 13q14.3.
METHODS and RESULTS: We performed FISH analyses of hematopoietic and non-hematopoietic cell lines to assess replication timing and chromatin packaging of the critical region. In line with an imprinting mechanism, we find that the two copies of the critical region replicate asynchronously and/or show delayed chromatid segregation, suggesting differential chromatin packaging of the two copies of 13q14.3. Next, we found by sequencing of SNPs that 13q14.3 candidate genes are expressed from one copy only in healthy probands. However, expression originated from either the maternal or paternal copy, excluding an imprinting mechanism. We could also show a functional interconnection of DNA methylation and gene expression, as demethylating agents and histone hyperacetylation induced biallelic expression. However, replication timing was not affected. Currently we are employing array- and capillary electrophoresis-based analysis of DNA-methylation (aPRIMES and bioCOBRA) and chromatin-immunoprecipitation on arrayed CpG-libraries (chIP on chip) with antibodies specific for histone modifications in order to identify the epigenetic element regulating the critical region.
CONCLUSIONS: We propose that differential replication timing represents an early epigenetic mark that distinguishes the two copies of 13q14.3, resulting in differential chromatin packaging and monoallelic expression. This has profound effects for the tumor suppressor mechanism localized in 13q14.3: Deletion of the single active copy of the region at 13q14.3, which is detected in more than 50% of CLL tumors, will suffice for complete loss of tumor suppressor function, as the remaining gene copies are epigentically silenced. In addition, we are currently identifying the locus control region that orchestrates gene expression in the critical region. Thus, we provide a model for the pathomechanism of 13q14.3 in CLL by the interaction of genetic lesions and epigenetic silencing.
Embryonic Stem Cells Transplanted In Utero Survive and Produce Teratocarcinomas in a Murine Model.
Authors: Nedelcu Elena , Srivastava Anand , Assatourian Gustavo , Varki Nissi , Carrier Ewa 
Published: October 2019

Abstract
Introduction: Embryonic stem (ES) cells have the capacity to differentiate into any cell type. We have previously established an in vivo murine model for the study of the in vivo differentiation of ES cells. The aim of this study was to monitor tumor formation in this model. Materials and
Methods: Murine ESC genetically engineered to express yellow fluorescent protein (YFP-ES cells) were cultured on mitomycin-treated feeder layers for four days prior to in utero transplantation in medium containing leukemia inhibitory factor. A cell dose of 5x104 YFP-ES cells (H-2b) was injected in utero intraperitoneally in the fetuses of Balb/c (H-2d) and C57BL/6 (H-2b) pregnant mice at E12- E14. The presence of YFP-ES cell-derived cells was analyzed in various tissues of the liveborn offsprings of different ages by fluorescence microscopy and quantified by digitalized fluorescence microscopy of frozen section from various tissues and organs and analysis of tissue supernatants for the presence of the YFP signal. Tumor formation was monitored in liveborn offsprings for up to one year and tissue sections were examined by routine and fluorescent microscopy.
Results: The YFP-fluorescent cells derived from YFP-ES cells were detected in liver and thymus without any evidence of migration in other organs. Extensive peritoneal teratocarcinomas with supradiaphragmatic involvement were generated in two out of five allogeneic in utero transplantation of YFP-ES cells. Tumor formation was not observed in the groups transplanted in utero with fetal liver or adult bone marrow hematopoietic cells, or in the control group. Clinical and histopathological pictures suggestive of graft-versus -host disease were also observed in two out of five mice transplanted in utero.
Conclusion: The promise and the potential risks of the ESC transplantation have to be carefully considered due to the high risk of teratoma/teratocarcinoma occurrence. Protocols using pre-differentiated cells from ES or adult stem cells may constitute a better approach for cellular therapies.
Cytotoxic Effects of CML28 Specific T Cells on Acute Leukemia Cells In Vitro.
Authors: Mao Hanwen , Liu Wenli , Gen Zhe , Huang Wei , Zhang Yicheng , Zhou Jianfeng , Sun Hanying 
Published: October 2019

Abstract
The antigen-specific cytotoxic T lymphocyte activated by antigen presenting cell is widely used in cell immunotherapy recently. CML28, which was screened from chronic myelogenous leukemia (CML) patients, was reported to be a specific tumour antigen and over-expressed on CML cells and acute leukemia cells. Therefore, CML28 could be a potential target for leukemia treatment. Dendritic cells (DC) are the most important antigen present cells, but it is hard to isolate and culture DCs for clinical use, which hampers the specific cell immunotherapy. Our investigation aimed to study the cytotoxic effects of CML28 specific T cells activated by artificial antigen presenting cells, on acute leukemia cells in vitro. Artificial antigen presenting cells were prepared by connecting CML28 to magnetic superbead that containing HLA-A2-Ig and B7-1 molecule. Mononuclear cells were isolated from the bone marrow or peripheral blood of healthy donors with positive HLA-A2. The artificial antigen-presenting cells were co-cultured with isolated mononuclear cells for four weeks. The activation and proliferation of CML28-specific T cells were measured by dimmer binding technique using flow cytometry. The cytotoxic effects of CML28-specific T cells on leukemia cells, which were isolated from leukemia patient, were evaluated by lactate dehydrogenase (LDH) releasing assay. Increased proportion of CML28-specific T cells was observed in artificial antigen-presenting group than in control group (29.27±3.54% vs 2.95±0.66%, p<0.05). For cytotoxic effects assay, significant higher killing efficiency was seen in artificial antigen-presenting group (41.47±4.23%vs3.56±0.71%, when the effector: target ration is 40:1, p<0.01). Therefore, we concluded that the artificial antigen presenting cells could mimic antigen presenting cells to induce specific T cell activation and proliferation, and cytotoxic effects on target cells, indicating that artificial antigen presenting cell-induced cytotocix T cells could be an option for leukemia treatment.
Recombinant Chimeric IgA1 and IgA2 Antibodies for Tumor Immunotherapy.
Authors: Dechant Michael , Beyer Thomas , Lohse Stefan , Schneider-Merck Tanja , Weisner Wencke , Berger Sven , Peipp Matthias , van de Winkel Jan G.J. , Valerius Thomas 
Published: October 2019

Abstract
The therapeutic potential of human IgA antibodies has hardly been explored, although IgA is the most abundantly produced antibody isotype in humans. Here, we describe the generation and functional characterization of human/mouse chimeric IgA antibodies against the epidermal growth factor receptor (EGF-R), which is a validated target antigen for tumor immunotherapy. Human IgG1, IgA1 and IgA2 variants of the C225 antibody were produced in CHO-K1 transfectomas using serum-free, non-adherent culture conditions. Productivity rates of selected IgA producing clones were approximately 5 pg/cell/day. A two-step purification procedure consisting of thiophilic agarose columns followed by gel electrophoresis on Superdex 200 yielded highly purified monomeric IgA antibodies, which were stable at 4° C. Silver staining and Western blots with human kappa-light- and human α-heavy chain antibodies confirmed the purity and correct assemby of IgA1 and IgA2 antibodies. Co-transfection of a His-tagged joining (J) chain significantly increased the amount of dimeric IgA antibodies, which were detected by anti-His antibodies. IgG1 and monomeric IgA antibodies demonstrated similar binding to EGF-R expressing A431 tumor cells (EC50 approx. 10 μg/ml) and to EGF-R transfectants. Furthermore, IgA1 and IgA2, but not IgG1 demonstrated binding to FcαRI (CD89)- transfected cells. All three isotypes were similarly effective in recruiting F(ab’)- mediated killing mechanisms such as blockade of EGF binding, inhibiting EGF-R phosphorylation, and mediating growth inhibition of EGF-R expressing cells. All three isotypes did not activate complement- mediated killing. Human IgG1 effectively triggered MNC- mediated ADCC, but activated PMN only weakly. Importantly, both human IgA isotypes were significantly more effective than IgG1 in recruiting neutrophils for ADCC (EC50 approx. 0.25 μg/ml), and triggered ADCC at effector to target cell ratios as low as 5:1. Since neutrophils constitute the most abundant effector cell population in human blood, IgA antibodies triggered higher levels of tumor cell killing in whole blood assays than human IgG1 antibodies - particularly when blood from G-CSF- primed donors was compared to healthy donor blood. Together, these data suggest that recombinant human IgA antibodies can be produced in reasonable amounts, and that they may constitute promising reagents for immunotherapy.
Immunostimulatory Properties of the Human Ortholog of the GMCSF/IL2 Fusion Protein for Cell Based Therapy.
Authors: Graduate Claudia Penafuerte , Galipeau Jacques 
Published: October 2019

Abstract
NK cells constitute a potential candidate for cancer cell therapy because they express a diverse array of inhibitory and activating receptors, which recognize and kill infected or tumor cells without prior immune sensitization. However, autologous NK cell mediated adoptive immunotherapy is restricted due to insufficient cytolytic activity of NK cells from patient with aggressive malignancies. In contrast, the infusion of alloreactive NK cells has shown more successful outcomes in the treatment of cancer, but this approach also presents difficulties such as the high doses of cytokines required to induce NK cell expansion ex vivo, which may also sensitize NK cells to apoptosis. Therefore, a critical issue for NK cell based therapy is the use of appropriate growth factors or cytokines that promote NK cell expansion and activation. We have previously shown that a murine GM-CSF/IL-2 fusion protein (aka GIFT2) displays novel antitumor properties in vivo compared to both cytokines in combination regarding tumor site recruitment of macrophages and significant functional NK cell infiltration [Stagg et al., Cancer Research (December 2004)]. In the present work, we found that human GIFT2 will lead to a substantial two fold proliferation of human blood-derived NK cells which is significantly (p<0.05) superior to either IL2 or GMCSF single cytokine treatment or both cytokines combined at equimolar concentration. In addition, we observed that GIFT2 leads to robust expression of NK-cell activation markers CD69 and CD107a. In conclusion, the human GIFT2 fusokine is a novel and potent tool for ex vivo expansion of activated NK cells which may be of use in cell-based immunotherapy of cancer.
KIR Incompatible NK Cells Effectively Lyse Osteosarcoma Cells.
Authors: Delgado David C. , Hardin Heather , Webster Dan , DeSantes Ken B. , Shaaban Aimen F. 
Published: October 2019

Abstract
Recurrent or metastatic pediatric solid malignancies, including osteosarcoma, carry a dismal prognosis despite modern multi-modality treatment approaches. Although the success of KIR (Killer Immunoglobulin-Like Receptor) incompatible, haploidentical stem cell transplantation has been documented in hematological malignancies in adults and children, this approach has not been thoroughly examined in pediatric solid tumors. In this study, we evaluated the potential for KIR-incompatible lysis of osteosarcoma cells, in vitro. We hypothesized that the killing of osteosarcoma cell targets could be predicted by the degree of inhibitory KIR receptor/ligand mismatch with NK cell effectors. To test this hypothesis, healthy donor NK cells were isolated by magnetic bead sorting and their KIR phenotype determined by flow cytometry. Consistent with previous studies, donor NK cells exhibited a high prevalence of all three relevant inhibitory KIRs (KIR2DL1, KIR2DL2/KIR2DL3, KIR3DL1). Conversely, examination of three established osteosarcoma cell lines (HOS, SaOS, and U2OS) demonstrated significant variability in cell surface HLA class I expression, by flow cytometry. QRTPCR analysis of these cell lines revealed variable expression of the three known inhibitory KIR ligands (HLA-C groups 1 and 2, HLA-Bw4). This variable KIR-ligand expression allowed evaluation of NK-mediated lysis of targets with varying KIR receptor-ligand incompatibility. Following a 12-hour incubation of donor NK cells in IL-2, lysis of osteosarcoma targets was measured in an annexin V flow cytometric assay. Osteosarcoma cell lines that expressed fewer KIR ligands consistently showed greater susceptibility to NK-mediated cytotoxicity. These findings were consistent using NK effectors from different donors. Additionally, we observed that at high passage number (>20), SaOS cells demonstrated down-regulation of KIR ligand expression. These changes correlated with increased lysis by the same donor NK cells. Our findings suggest:
Variable expression of KIR ligands in osteosarcoma cell lines allows potential susceptibility to KIR-incompatible, NK cell-mediated lysis; The killing of osteosarcoma cells by NK cells can be predicted by the degree of receptor/ligand mismatch; and During expansion, osteosarcoma cells may alter expression of KIR ligands, resulting in increased or decreased susceptibility to lysis by KIR-incompatible NK cells.
Further studies are needed to explore the utility of KIR-incompatible, haploidentical stem cell transplantation for patients with high-risk osteosarcoma.
Methylenetetrahydrofolate Reductase (MTHFR) Polymorphisms of Both Donor and Recipient Can Modify the Rate of Recurrence in Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Oh So Yeon , Kim Hye Jin , Yi Hyeon Gyu , Kim Jin Won , Piao Cheng Zhe , Chang Chongwon , Shin Eunsoon , Kim Inho , Yoon Sung-Soo , Park Seonyang , Kim Byoung Kook 
Published: October 2019

Abstract
Introduction Clinical significance of polymorphism in MTHFR gene is widely described in various disease entities. Polymorphism is associated with decreased metabolism of methotrexate. Because methotrexate is commonly used to prevent GVHD in HSCT, we studied transplantation outcomes according to genotypes of donor and recipient MTHFR; about the most studied two polymorphism sites (677C/T and 1298A/C). Materials and Methods Patients and donors who underwent allogeneic HSCT due to hematologic malignancy between 1998 and 2006 were retrieved. Patients had received MTX and cyclosporine for GVHD prophylaxis were included. The genotyping of MTHFR polymorphism was screened using SNaPShot Multiplex kit. Clinical data were collected retrospectively. Results We found 99 patients consisted of 60 males (60%) and 39 females (39%) in this homogeneous population. Median age was 34 years (range17–59). Diagnosis were acute leukemia (n=67), chronic leukemia (n=20) and other diseases (n=12). Donor was related(n=72) or unrelated(n=27). Conditioning was myeloablative in 95 patients and nonmyeloablative in 4 patients. Median PFS and OS were 6 (range 0–101) and 15 (range 0–108) months. Clinical factors and genotype are analyzed for risk of hepatic toxicity, recurrence, TRM and death. Peak AST and ALT levels are higher in the 677TT group than in the CC or TT group. In univariate analysis, recipient or donor 1298AA, AC, and CC were likely to relapse in ascending orders (figure1). This was reproduced in multivariate analysis (p=0.008 and 0.011, for each). About TRM and survival, there were no significant correlations with MTHFR genotypes in both univariate and multivariate analysis. Conclusion Simultaneously decreased activity of MTHFR due to 1298AC polymorphism of both donor and recipient might be translated into increased recurrence rate. Validation with randomized trial is required to compare methotrexate to other drug for GVHD prophylaxis.
Figure Figure
Results of a Randomized Phase II Trial of Pegfilgrastim Versus Filgrastim To Treat Neutropenia Post-Autologous Peripheral Blood Stem Cell Transplant (PBSCT) in Patients with Non-Hodgkin’s Lymphoma (NHL).
Authors: Rifkin Robert , Beveridge Roy , Spitzer Gary , Orloff Gregory , Mandanas Romeo , McGaughey Dean , Brunvand Mark , Zhan Feng , Boehm Kristi A. , Asmar Lina 
Published: October 2019

Abstract
Granulocyte colony-stimulating factor has been shown to decrease the time to neutrophil recovery following autologous PBSCT. Therefore, it was hypothesized that a single injection of pegfilgrastim (P) would mimic the role of filgrastim (F), resulting in at least an equivalent shortening of post-PBSCT neutropenia. PBSCT eligible NHL patients over the age of 18 years with preserved end-organ function were identified prior to the administration of high-dose chemotherapy, and following adequate stem cell harvest and cryopreservation (ie, >2.5 x 106 CD34+ cells/kg). All patients received either standard BEAM or BEAC high-dose chemotherapy (HDC). Prior to HDC, patients were randomly assigned to receive either P at a fixed-dose of 6 mg on Day +1(Arm A), or weight-based, dose-adjusted F rounded to the nearest prefilled syringe beginning on Day +1(Arm B) following transplantation. Between July 2003 and April 2007, 101 eligible patients were enrolled within our transplant network. Three patients were deemed ineligible (CHF death, patient withdrew consent, other). The analyses and results presented below outline the remaining 98 patients. The demographic characteristics of both arms were well-balanced with regards to stage at diagnosis, stage at treatment, ECOG Performance Status, histology, and prior therapy. The comparison of P vs F is summarized in the table below. Incidence of Grade 3–4 adverse events were comparable in both arms as was transplant-related mortality. In conclusion, administration of pegfilgrastim post-auto PBSCT appears to be equivalent to multiple daily dosing of filgrastim, and might be considered in lieu of filgrastim obviating the need for multiple injections.
Results - Comparison of Pegfilgrastim vs Filgrastim Variable Arm A (P) Arm B (F) No. of Patients Treated 50 48 Doses Received (mean +/− SD 1.0 +/− 0 12.7 +/− 2.6 Time to ANC Recovery (days) (mean +/− SD) 8.3 +/− 1.1 8.9 +/− 1.5 No. of RBC Transfusions (mean +/− SD) 1.7 +/− 0.9 1.9 +/− 1.2 No. Platelet Transfusions (mean +/− SD) 3.0 +/− 1.9 2.8 +/− 1.8 Positive Blood Culture Rate (%) 18.0% 29.0% Febrile Neutropenia (FN) Rate (%) 18.0% 16.7% Duration of FN (days) (mean +/− SD) 5.1 +/− 3.4 5.5 +/− 4.9