bluuebunny/crossref_metadata_2025_split huggingface dataset:
GlycoPEGylated Filgrastim XM22 Demonstrates Dose-Dependent Activity in Comparison to Pegfilgrastim in a Dose-Escalating Study in Healthy Volunteers after Body-Weight Dependent Single Dose.
Authors: Lubenau Heinz , Pokorny Rolf , Lopez-Lazaro Luis , Bias Peter , Wallin Bruce , Hinderer Walter , Maly Anne-Katrin , Seiberling Michael 
Published: October 2019

Abstract
XM22 is a glycoPEGylated recombinant human G-CSF that interacts with the G-CSF receptor by binding and activation. In pilot studies in non-neutropenic animals, XM22 demonstrated a similar PK/PD profile compared to pegfilgrastim. Because of its longer duration of action compared to filgrastim, XM22 is intended to be dosed once per cycle for the reduction of duration of severe neutropenia and incidence of febrile neutropenia in cancer patients undergoing chemotherapy.
METHODS: A single center, randomized, single-blind study of XM22, administered as single escalating body-weight (bw) dependent doses given by the s.c. route was conducted to assess safety, pharmacokinetic profile and pharmacodynamic properties based on absolute neutrophil count (ANC) and CD34+ cell count. Male or female healthy volunteers were assigned to one of 3 XM22 dose groups, at 25 μg/kg bw dose (n=8), at 50 μg/kg bw (n=15), and 100 μg/kg bw (n=15) compared to 100 μg/kg bw pegfilgrastim (n=15).
RESULTS: 53 healthy volunteers were enrolled in the three dose groups and completed 21 day follow-up. Injections were generally well-tolerated with no discontinuations due to adverse events or serious adverse events. XM22 at a dose of 100 μg/kg bw exhibited a PK profile with increased bioavailability and extended half life compared to the same dose of pegfilgrastim. ANC area over baseline effect curve (AOBEC) and CD34+ AOBEC were higher after treatment with XM22 100 μg/kg bw (6818.55 hr*neut./nl and 7340.16 hr*cell count/μl geometric mean respectively) compared to the same dose of pegfilgrastim. Antibody data will be presented.
CONCLUSIONS: Single body-weight dependent doses up to 100 mg/kg of XM22 administered to healthy volunteers were generally well-tolerated with regard to the expected side effect profile and demonstrated dose-dependent increases in absolute neutrophil count and CD34+ cell count. These data supplement results from another study in healthy volunteers in which XM22 was administered as a fixed dose.
New Classification of Chronic Graft Versus Host Disease: Added Clarity from the Consensus Diagnoses.
Authors: Arora Mukta , Nagaraj Sriharsha , Witte Judy , DeFor Todd E. , MacMillan Margaret L. , Burns Linda J. , Weisdorf Daniel J. 
Published: October 2019

Abstract
Chronic graft versus host disease (CGVHD) is classically defined as occurring more than 100 days after hematopoietic cell transplant (HCT). But, with newer donor types and transplant strategies being used [eg. non myeloablative (NMA) and reduced intensity conditioning (RIC)] the presentation of the syndrome is changing. Clinical syndromes resembling acute GVHD (aGVHD) are often recognized beyond 100 days after HCT; possibly misdiagnosed as CGVHD. Additionally, the old grading system of limited versus extensive GVHD was designed to identify patients likely to benefit from systemic immune suppression and does not capture the severity of organ involvement. The Diagnosis and Staging Working Group of the NIH Consensus Development Project on Criteria for Clinical Trials in chronic graft-versus-host-disease proposed criteria for diagnosis and assessment of overall CGVHD severity. We retrospectively reviewed 197 patients who underwent sibling donor transplant between January 2002 to December 2005 to assess the applicability of the new criteria in predicting survival and transplant related mortality (TRM). Fifty four patients were diagnosed with CGVHD [cumulative incidence 28% (95% CI 22–34%)]. Of these, 8 [cumulative incidence 4% (95% CI 2–6%)] were reclassified as Late aGVHD; 31 [cumulative incidence 16% (95% CI 11–21%) had classic CGVHD and 15 [cumulative incidence 8% (95% CI 5–11%] had Overlap syndrome. Forty six patients with Overlap syndrome and classic CGVHD were graded. Six patients (13%) had Mild, 29 (63%) Moderate and 11 (24%) Severe CGVHD. After a median follow up of 35 months (range 4.3–59.2mo), 3 year probability of overall survival was significantly worse in Late aGVHD [25%(95% confidence interval 4–56%)] as compared to Overlap [87%(56–96%)] or CGVHD [75%(54–87%), p=0.001].). Amongst patients with Overlap and CGVHD (n = 46), similar survival was seen between mild or moderate disease. However, patients with severe disease had a trend to worse survival. [Mild-moderate 3 year probability of survival 85% (68–94%) versus Severe 57%(21–82%)]. Additonally, 3 year cumulative incidence of TRM was higher in patients with Late aGVHD [38% (9–67%)] versus Overlap syndrome [13%(0–27%)] or CGVHD [7%(0–13%)], p= 0.003. A trend towards greater TRM was seen in patients with severe GVHD [18%(0–36] versus mild+ moderate GVHD [6%(0–14%)]. This analysis indicates that the consensus guidelines are applicable to related donor HCT recipients and more reliably predicts survival and TRM than older classification of CGVHD. Prospective validation and testing in alternative donor cohorts is still needed before widespread acceptance is justified.
The Optimal Processing Time of Adipose-Derived Mesenchymal Stem Cell after Lipoaspiration.
Authors: Jang Jun Ho , Kim Kihyun , Jung Chul Won , Park Keon Woo 
Published: October 2019

Abstract
Background: Our aims were to characterize the phenotype of these cells, to investigate their immunoregulatory properties in vitro, and to determine the optimal processing time of AD-MSC after lipoaspiration.
Methods: ASCs were isolated from lipoaspirated adipose tissues by treatment of collagenase A and cultured in a Dulbecco’s Modified Eagle’s Medium (DMEM). To evaluate the optimal processing time of ASCs, the nucleated cells were processed by collagenase treatment at 3, 6, 9, 12, 15, 24, 30 and 36 hours after lipoaspiration respectively. To know the characteristics of ASCs, the expression of cell surface antigens was analyzed by flow cytometry, and the proliferation potentials were estimated by colony forming abilities or capacities of population doubling. The differentiation potentials into adipocytes or osteoblasts were confirmed by accumulation of neutral lipid vacuoles stained with Oil-red O and expression of alkaline phosphatases.
Results: When the nucleated cells were isolated by collagenase treatment after lipoaspiration, the mean cell yield was about 3.1 × 106 or 1.2 × 106cells per gram of lipoaspirate (n=8) processed. Nucleated cell counts were increased with time but plateau between 15h and 24h after initial processing and decreased after 24h. Flowcytometric analysis showed that Adipose tissue-derived stem cells (ASCs) have a marker expression that is similar to that of bone marrow stromal cells (BMSCs). ASCs expressed CD44, CD73, CD90, and CD105 and were absent for CD14, CD31 and CD45 expression. When primary cells were plated at 50 or 1000 cells/cm2 on 6-well plates, cumulative population doublings were about 50 times until passage 7 or 13 (approximately 130 days), respectively, and ASCs expanded to 1018 cells. ASCs were multipotent, differentiating along the adipocyte and osteoblast lineages. ASCs did not provoke in vitro alloreactivity of incompatable lymphocytes and, moreover, suppressed mixed lymphocyte reaction (MLR) and lymphocyte proliferative response to mitogen.
Conclusion: The optimal processing time for AD-MSCs from lipoaspirates should before 24h after removal RBC and AD-MSCs can be a alternate source of multilineage MSCs for clinical use such as tissue repair and transplantation.
GM-CSF Stimulates IL-8 Expression in Human Bone Marrow Stromal Cells Via Activation of NADPH Oxidases.
Authors: Wu Shuling , Enders Sven , Burmeister Thomas , Nowak Daniel , Reinwald Mark , Thiel Eckhard 
Published: October 2019

Abstract
The self-renewal, proliferation, and mobilization of hematopoietic progenitor cells (HPC) depend on complex interactions involving cytokines and bone marrow (BM) stromal microenvironment. Little is known about the effect of mobilizing agents such as granulocyte-macrophage colony-stimulating factor (GM-CSF) on BM stromal cells and their contribution to the mobilization of peripheral blood stem cells. In the present study we show that, 3 hours after addition, GM-CSF distinctly upregulated IL-8 mRNA expression and protein secretion in cultured human BM stromal cells. Furthermore, GM-CSF stimulated the generation of reactive oxygen species (ROS) in a dose- and time-dependent manner. Intracellular levels of ROS correlated with the extent of IL-8 production and IL-8 promotor activity as determined by luciferase reporter assay. Using western blot analysis, we showed that BM stromal cells expressed both phagocytic and non-phagocytic NAD(P)H oxidase components such as gp91phox, NOX1, p22phox, p47phox, p67phox, and NOXO1, suggesting that the upregulation of IL-8 expression induced by GM-CSF may be associated with the activation of stromal cell NADPH oxidase activity. GM-CSF directly stimulated the phosphorylation of p47phox, a key event in NAD(P)H oxidase activation. Inhibitors of the ERK1/2 and AKT pathways abrogated GM-CSF-induced phosphorylation of p47phox, indicating that GM-CSF-induced phosphorylation of p47phox is mediated by these pathways in BM stromal cells. Accordingly, the transactivation of IL-8 promotor activity induced by GM-CSF was suppressed by these inhibitors and the antioxidants diphenylene iodonium (DPI) and N-acetyl-l-cysteine (NAC). In conclusion, these findings show that GM-CSF elicits the activation of the redox-sensitive signaling via ROS, resulting in stromal cell function such as cytokine production and therefore possibly impacts on hematopoiesis.
Phase 1 Single Agent Antitumor Activity of Twice Weekly Consecutive Day Dosing of the Proteasome Inhibitor Carfilzomib (PR-171) in Hematologic Malignancies.
Authors: Alsina Melissa , Trudel Suzanne , Vallone Marcy , Molineaux Christopher , Kunkel Lori , Goy Andre 
Published: October 2019

Abstract
Carfilzomib (CFZ) is a structurally- and mechanistically-novel proteasome inhibitor of peptide epoxyketone class that exhibits a high level of selectivity for the unique N-terminal threonine active sites within the proteasome. CFZ is similar to bortezomib (BTZ,Velcade®), in that it is a potent inhibitor of the proteasome chymotrypsin-like activity, but unlike BTZ, CFZ has shown minimal cross-reactivity with the other catalytic sites within the proteasome or across other protease classes. Preclinical human tumor xenograft models indicated that consecutive day (D1, D2) dosing resulting in 48 hours of proteasome suppression was superior to split dosing (D1, D4). This phase 1 sequential dose escalation trial tested twice weekly consecutive day dosing (48 hour proteasome inhibition) with carfilzomib. Carfilzomib was administered as an IVP on a 28 day cycle (D1, 2, 8, 9, 15 and 16 with 12 days rest) at doses from 1.2 mg/m2 to 27 mg/m2. Patients with Multiple myeloma (MM), non-Hodgkin’s Lymphoma (NHL) that included mantle cell lymphoma (MCL), Hodgkin’s disease (HD), or Waldenström’s macroglobulinemia (WM) were eligible if they had relapsed after at least 2 prior therapies. A total of 37 subjects have been treated. The minimal effective dose (MED) was 15 mg/m2, and maximum (80%) proteasome inhibition was achieved in peripheral blood and mononuclear cells at this dose. Five objective responses have occurred in 16 patients (13 MM, 3 MCL) enrolled at ≥ the MED: 4 MM partial response (PR), 1 MM minimal responses (MR). The responses have been durable (134 to 392 days) and occurred in patients who had failed bortezomib, immunomodulatory agents and stem cell transplant. Six additional patients have had stable disease (SD): 2 MM, 1 T NHL, 2 MCL, 1 B NHL. Dose limiting toxicity reported at 27 mg/m2 included a hypoxic event and Grade 2 thrombocytopenia increasing to Grade 4. A reversible Grade 2 creatinine D2 was reported in 3/5 MM patients treated at 27 mg/m2 and was associated with a rapid decline in M-protein without evidence of tumor lysis syndrome and the event did not occur on rechallenge. Cyclic thrombocytopenia was rapidly reversible and painful peripheral neuropathy was not reported. This study demonstrates that consecutive CFZ dosing leading to sustained proteasome inhibition is well tolerated on a weekly basis. The selectivity of epoxyketone class of proteasome inhibitors may provide a wide therapeutic index. Patients have remained on therapy for over a year, and furthermore, antitumor responses in heavily pretreated MM patients suggest non-cross resistant activity. Phase 2 trials in relapse and refractory MM and relapse solid tumors are proceeding with this schedule.
Functional Alterations of Lin−CD34+CD38+ Progenitors in Chronic Myelomonocytic Leukaemia and on Progression to Acute Leukaemia.
Authors: Sun Qian , So Chi-Chiu , Yip Sze-Fai , Wan Thomas S.K. , Ma Edmond Shiu Kwan , Chan LiChong 
Published: October 2019

Abstract
Chronic myelomonocytic leukaemia (CMML) is a clonal bone marrow stem cell disorder based on the presence of trilineage involvement, the association of myelodysplastic and myeloproliferative features and its ability to transform into acute myeloid leukaemia. The objectives of our study are to identify the cell population and its functional characteristics involved in evolution from CMML phase to acute myeloid leukaemia. We analysed Lin−CD34+ stem/progenitor population and performed cell proliferation, apoptotic assays, self-renewal ability and differentiation potential studies in purified populations of Lin−CD34+CD38− stem cells and Lin−CD34+CD38+ committed progenitors from peripheral blood of 16 patients with CMML and in six of the 16 after transformation to acute myeloid leukaemia (AML-t). We observed an expansion of the stem cell/progenitor pool (Lin−CD34+ cells) in AML-t comprising mainly of Lin−CD34+CD38+ committed progenitors within Lin−CD34+ cells. The Lin−CD34+CD38+ committed progenitors displayed highly proliferative activity in CMML and in AML-t; and additionally acquired resistance to apotosis and myeloid colony self-renewing ability in AML-t. Impairment of dendritic cell (DC) differentiation was observed with complete block in AML-t. Our findings suggest Lin−CD34+CD38+ committed progenitors instead of Lin−CD34+CD38− stem cells could be the target(s) of secondary genetic lesions underpinning progression from CMML to AML. These results have implications for the further study of the biology of leukaemic transformation and the design of new strategies for the effective treatment of CMML.
The Establishment and Characterization of a Human Acute Myelocytic Leukemia Cell Line, SH-2 Carrying t(16;17))(q24;q12) Translocation.
Authors: Qiu Huiying , Xue Yongquan , Pan Jinlan , Wu Yafang , Wang Yong 
Published: October 2019

Abstract
We report the establishment of a novel cell line from a patient with acute myelocytic leukemia. The initial growth of this cell line was dependent on the presence of the stromal cell and cytokine interleukine 3. Subsequently, a stroma and cytokine-independent cell line was established, named SH-2. The cell line has proliferated continuously in vitro for more than 24 months. The SH-2 cell line showed identical chromosomal alterations to that of the initial bone marrow aspiration specimen, demonstrated a t(16;17))(q24;q12) translocation accompanied by deletions of y and 17 and triple 19.Chromosome painting FISH and Multi-FISH conformed the karyotype.SH-2 cells expressed myeloid antigens CD13, CD33 and MPO, SH-2 co-expressed NK markers CD16 and CD56. The loss of one p53 allele were proven by chromosome painting, FISH. A point mutation of CAG to CAT at codon 576 of extron 5 in another p53 allele was found by direct sequencing of DNA. Neither Epstein-Barr virus nor mycoplasma was not detected in SH-2 cells. DNA fingerprinting confirmed the authenticity of the cell line. Cytokines of GM-CSF,IL-3,IL-6,SCF,IL-4 and IL-5 can improve the proliferation of SH-2. SH-2 has colony formality and tumorigenic capacity in nude mice. The breakpoints of chromosomes 16 and 17 were localized RP11-356C4(16q24.3)and between BAC clone RP1-161P9 and RP11-47L3 corresponding to 17q12. The establishment of an myelocytic leukemia cell line with t(16;17)(q24;q12) could be valuable for the study of leukemogenesis and for the research of cloning the new gene involved in the t(16;17)(q24;q12) translocation.
Preferential Hypermethylation of the DKK-1 Promoter in Core-Binding Factor Leukaemia.
Authors: Suzuki Rikio , Onizuka Makoto , Kojima Minoru , Shimada Masako , Fukagawa Satomi , Tsuboi Kosuke , Kobayashi Hiroyuki , Shintani Ayumi , Ogawa Yoshiaki , Kawada Hiroshi , Hotta Tomomitsu , Ando Kiyoshi 
Published: October 2019

Abstract
The Dickkopf-1 (DKK-1) gene product is an extracellular Wnt inhibitor. Hypermethylation of the DKK-1 promoter results in transcriptional silencing and may play an important role in cancer development. Here, we investigated hypermethylation of the DKK-1 promoter in patients with acute myeloid leukaemia (AML), especially core-binding factor (CBF) leukaemia. The methylation status of DKK-1 was analyzed by methylation-specific polymerase chain reaction in 47 patients with AML. DKK-1 methylation was found in 14 (29.8%) of the patients, and more frequently in those with CBF leukaemia (6 of 12 patients), than in those with acute promyelocytic leukaemia (0 of 6 patients) (P = .03). In contrast, Wnt inhibitory factor-1 methylation was found in acute promyelocytic leukaemia (4 of 6 patients) but not in CBF leukaemia (0 of 12 patients) (P = .001). Multivariate analyses suggested that DKK-1 methylation was a risk factor for poorer overall survival. Sequential analysis using 4 paired samples obtained at diagnosis and relapse suggested that DKK-1 methylation was involved in the progression of leukaemia. DKK-1 methylation may therefore be involved in leukaemogenesis, especially in CBF leukaemia, and may be a useful prognostic marker in AML.
The Epigenetic Effect of Phenylhexyl Isothiocyanate on Histone Acetylation in Patients with Acute Leukemia and Non-Leukemic Disorders.
Authors: Xiao Liyun , Ma Xudong , Huang Yiqun , Zhen Ruiji , Ye Xiaoling , Chiao Jen-Wei , Liu Delong 
Published: October 2019

Abstract
Phenyhexyl isothiocyanate (PHI) has been shown to inhibit histone deacetylation in leukemia cell lines in vitro and in animal models. In this study we examined the histone acetylation status in leukemia cells from patients with acute leukemia as well as in non-leukemia cells. From January to September 2006, 19 patients with newly diagnosed acute leukemia were enrolled. 12 patients had AML, 7 had ALL. The age ranged between 2 months to 65 years old. There were 12 males and 7 females among them. 5 non- leukemia patients were also enrolled, with 1 anemia, 1 infection disease, 2 ITP, and 1 healthy volunteer. Mononuclear cells were isolated from patients’ specimen. Apoptosis of the cells and histone acetylation of H3 and H4 were characterized pre- and post- PHI treatment. The expression of acetylated H3 and H4 in acute leukemia cells was decreased, as compared to those in non-leukemia cells. Increased apoptosis was seen in the acute leukemia cells after PHI treatment of the primary culture of the leukemia cells for 3 and 7 hours. The effect of PHI on the leukemia cells from patients was time- and dose-dependent. There was a significant increase of PHI effect on leukemia cells versus non-leukemia cells (p<0.05), suggesting that PHI may have preferential activity against leukemia cells. This study presents the first evidence of PHI with preferential activity against leukemia cells over non-leukemia cells from human subjects. Clinical trial is warranted to study PHI in leukemia patients.
Decreased Expression of the Histone Methyltransferase SUV39H1 in AML Cells Reactivates Hypermethylated Tumor Suppressor p15INK4B in the Absence of Promoter Demethylation.
Authors: Lakshmikuttyamma Ashakumary , Scott Stuart , Geyer C. Ronald , DeCoteau John F. 
Published: October 2019

Abstract
Re-expression of hypermethylated tumor suppressor genes using epigenetic modifiers, such as DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors, occurs by a mechanism whereby promoter demethylation is the dominant event. In support of this model, we found that the DNMT inhibitor 5-Aza-2-deoxycytidine (decitabine) induces expression of the tumor suppressor gene p15INK4B (p15) in AML cells with hypermethylated p15 promoters. Re-expression of p15 by decitabine is associated with decreases in p15 promoter methylation and histone H3 lysine 9 (H3K9) methylation and increases in H3K9 acetylation. DNA methylation is linked to H3K9 methylation through the DNA methyl binding protein MeCP2, which associates with DNMTs and H3K9 methyltransferases. Using chromatin immunoprecipitaton (ChIP) assays, we confirmed that MeCP2, DNMT1 and the H3K9 methylatransferase SUV39H1 interact with the methylated p15 promoter and that this interaction is reduced by decitabine. To determine whether promoter demethylation is also dominant to H3K9 demethylation, we monitored p15 re-expression in the presence of SUV39H1 shRNA alone and in combination with decitabine. SUV39H1 shRNA induces p15 expression and H3K9 demethylation, however it does not affect p15 promoter methylation. These results are in contrast to the HDAC inhibitor trichostatin A (TSA), which cannot induce p15 re-expression. SUV39H1 shRNA induced p15 expression and H3K9 demethylation are also enhanced by co-treatment with decitabine or TSA. Surprisingly, co-treatment with decitabine and SUV39H1 shRNA partially reverses decitabine induced promoter demethylation. Using ChIP assays we show that SUV39H1 shRNA increases the amount of the histone H3K9 dimethytransferase G9a and DNMT1 associated with the p15 promoter. Increased levels of G9a at the p15 promoter would enhance promoter methylation since G9a stimulates DNMT1 activity. Our results demonstrate that hypermethylated p15 can be reactivated in AML cells by an initial event that involves H3K9 demethylation. In addition, we found that the SUV39H1 inhibitor chaetocin induces p15 in AML cells with hypermethylated p15 promoters. Therefore, H3K9 methylatransferases represent novel therapeutic targets for developing inhibitors to reactivate the expression of hypermethylated genes.