bluuebunny/crossref_metadata_2025_split huggingface dataset:
Erythroid Kruppel-Like Factor Regulates E2F4 and the G1 Cdk Inhibitor, p18.
Authors: Perkins Andrew C. , Keys Janelle R. , Hodge Denise J. , Tallack Michael R. 
Published: October 2019

Abstract
Erythroid Kruppel-Like Factor (EKLF) is a zinc finger transcription factor which is essential for β-globin gene expression. Knockout mice die from anemia at E15, but restoration of globin chain imbalance does not rescue anemia or increase survival. Cell lines derived from EKLF null mice undergo proliferation arrest upon reactivation of a conditional EKLF-ER fusion protein, suggesting a role in cell cycle control. A transcriptional profiling experiment comparing the global gene expression in EKLF null and wild type fetal liver identified many differentially expressed genes, a number of which function in G1 and at the G1/S checkpoint of the cell cycle. The Cyclin dependent kinase (Cdk) inhibitor, p18, and the S phase transcription factor E2F4 were both found to be significantly down regulated in EKLF null mice and this result was confirmed by real-time PCR. Interestingly, E2F4 knockout mice have a similar phenotype to EKLF knockout mice. Bioinformatic searches of the p18 and E2F4 genes shows that each contains phylogenetically conserved CACC box motifs capable of binding EKLF within longer regions of conservation in promoter and intron regions. The p18 gene contains two conserved CACCC sites upstream of the start of transcription, which are required for EKLF dependent promoter activity in luciferase reporter assays. The transcription factor E2F4 contains a conserved EKLF-binding CACC site within an intron that is closely associated with two conserved GATA1 binding sites. We show by a chromatin immunoprecipitation (ChIP) assays that the E2F4 intron and p18 promoter are occupied by EKLF in vivo. Together, these results suggest that EKLF is likely to directly regulate expression of key cell cycle genes in vivo to drive the switch from proliferation to differentiation of erythrocytes. The loss of EKLF is likely to result in aberrant proliferation and predisposition to leukemia.
Cytoplasmic Tail of Tissue Factor(TF): Role in TF Trafficking to Microparticles.
Authors: Zakarija Anaadriana , Hobbs Jennifer , Chalasani Pavani , Lieberman Elliot , Morrissey James H. , Soff Gerald A. 
Published: October 2019

Abstract
Introduction: Tissue factor (TF) is a transmembrane protein that is primarily extracellular. It has a 21 amino acid cytoplasmic tail, which is not essential for coagulation but may have a role in signal transduction. We studied the activity of deletion variants of TF, transfected into a pancreatic cancer line (MiaPaCa-2), which lacks TF RNA or protein.
Methods: TF variants: lctTF (lacking cytoplasmic tail), sTF(soluble, lacking transmembrane and cytoplasmic domains), wtTF (wild type) were transfected into MiaPaCa-2 cells. Microparticles were prepared from serum-free conditioned media(CM) by ultracentrifugation at 100,000g. Presence of TF antigen was detected by immunoblot using a monoclonal antibody to TF, 10H10. Tissue factor activity (TFA) was assessed by recalcification time.
Results: Immunoblot confirmed the presence of TF in the cell lysates of MiaPaCa-2 cells transfected with wtTF or lctTF. In the sTF MiaPaCa-2 cells, TF was not detected in the cell lysates, but was present in CM. TF was present in CM of lctTF-transfected cells, but only trace antigen was observed in CM of wtTF cells. TF was microparticle-associated in the CM collected from lctTF cells, but not in the sTF cells. The TF protein made by sTF cells was not functional in the CM as demonstrated by no difference in TFA as compared to untransfected MiaPaCa-2 cells (>350 sec vs 351 sec). The CM from lctTF MiaPaCa-2 cells shortened clotting time by over 50% (159 sec vs 351 sec), while wtTF MiaPaCa-2 cells did not (349 sec vs 351 sec). The microparticles from lctTF had the most TFA when compared to wtTF or untransfected cells (107 sec vs 194 sec vs 436 sec). In the CM from cells with lctTF, TFA was exclusively in the microparticle section and absent in the supernatant (107 sec vs 440 sec).
Conclusions: Wild type TF remains primarily cell associated. In contrast, a higher fraction of the lctTF was released into the CM and remained microparticle associated. TF protein that lacks the transmembrane portion is soluble but not microparticle-associated and does not have procoagulant activity. Lack of the cytoplasmic tail results in increased generation of functional, microparticle-associated TF compared with the wild-type protein. These data suggest the cytoplasmic tail may influence the trafficking of functional TF to the microparticle fraction.
Novel Synthetic Histone Deacetylase Inhibitor (SK-7041) Potently Inhibits Proliferation in Acute Myeloid Leukemia Cell Lines.
Authors: Kim Young-Ju , Guo Susan , Park Ju Won , Bae Eun Kyung , Ahn Kwang-Sung , Kim Inho , Lee Jong-Seok , Lee Young Yiul , Park Seonyang , Kim Byoung Kook , Yoon Sung-Soo 
Published: October 2019

Abstract
Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities determine the acetylation status of histones, and have the ability to regulate gene expression through chromatin remodeling. Aberrant histone acetylation is known to play a key role in leukemogenesis. To improve the treatment outcome in AML patients, we examined the antiproliferative activity of SK-7041, a novel hybrid synthetic HDACI, in acute myelogenous leukemia (AML) cell lines (KG-1,HL-60 and HEL). This HDACI preferentially inhibited the enzymatic activities of HDAC1 and HDAC2, compared to other HDAC isotypes, indicating that class I HDAC is the major target of SK-7041. SK-7041 effectively induced the time-dependent hyperacetylation of histones, H3 and H4 in AML cell lines. We found that this compound exhibited potent antiproliferative activity against AML cell lines in vitro. The growth inhibitory effect of SK-7041 was related to the induction of apoptosis in AML cell lines. SK-7041(4uM) induced potent cell death, triggering apoptosis in over 60% of cells in 48h by FACS analysis. After 12h of treatment by western analysis, apoptotic cells were increased through activation of caspase-7,-9,-3 and cleaved-PARP (caspase-8 was not affected). Cyclin D1 expression was decreased, and CDK4 expression level was not affected by SK-7041. The level of p21 expression was gradually increased in KG-1 and HL-60 cell lines, whereas treated HEL cell lines showed no change. For further investigation of the genes affected by SK-7041, 20Kb oligonucleotide microarray analysis was performed. The expression of candidate genes, which we selected in chip analysis, was confirmed using real-time PCR. Fifteen genes (RGL1, FYN, CARD9, ABCA7, TNFRSF6B, CASP9, ENPP2, etc.) were consistently up-regulated and twelve genes (PTPN7, CD34, INSIG1, IL16, LHX6, TRIB3, BID, PDCD4, etc.) were down-regulated more than 4-fold by the SK-7041 compound. We suggest that the efficacy of SK-7041 was mainly mediated by the inhibition of proliferation and induction of apoptosis. Taken together, our data suggested that novel HDACI SK-7041 treatment is effective in AML cell lines, and that the further study is warranted.
Mesenteric Castleman’s Disease with Systemic Symptoms in Children.
Authors: Kim Sun Young , Sul Ji Young , Kim Jin Man , Chung Nak Gyun , Cho Bin , Jeong Dae Chul , Kim Hack Ki 
Published: October 2019

Abstract
Castleman’s disease (CD) is a lymphoproliferative disorder of unknown origin and rarely occurs in children. It has been further classified into two major subgroups: solitary or localized and multicentric CD. Clinically, hyaline-vascular type is rarely associated with systemic symptoms, but the plasma cell type is frequently associated with the constitutional symptoms of fever, malaise, night sweat and the abnormal laboratory markers. We encountered a case of the hyaline-vascular type CD located in the mesentery with systemic symptoms. She admitted suffering from prolonged fever, anemia, poor weight gain, dyspnea, and abdominal distension. Labaroatory data showed that microcytic hypochromic anemia, polyclonal gammaglobulinemia, hyperfibrinogenemia, elevated C-reactive protein, and increased IL-6 level. Computed tomography confirmed that the mass was located in the mesentery with multiple lymphadenopathies. The clinical and biochemical abnormal findings improved after surgical resection of the involved lymph node. Currently she has no signs of recurrence at 12 months postoperatively.
High Catalase Activity: Another Possible Pathway for Imatinib-Resistant CML Patients.
Authors: Luque Diogo G. , Ferreira Ana Carolina S. , Vasconcelos Flavia C. , Maia Raquel C. , Kwee Jolie K. 
Published: October 2019

Abstract
Imatinib (imatinib mesylate: Novartis Pharmaceuticals, Basel, Switzerland) is a well-known selective inhibitor of BCR-ABL tyrosine kinase activity, a hallmark enzyme in the pathogenesis of chronic myeloid leukemia (CML). Resistance to imatinib in CML patients has been associated with different and complex types of mechanisms ranging from nonspecific multidrug resistance protein, preventing the drug to reach its target, to BCR-ABL kinase domain point mutations, reducing the affinity of the enzyme to Imatinib. More recently, it was described that modulation of reduced glutathione (GSH) intracellular levels restores sensitivity to Imatinib in Imatinib-resistant cell line. This suggests that induction of hydrogen peroxide (H2O2), a well-known apoptotic reactive oxygen species (ROS), might be another Imatinib anti-leukemic mechanism action. Additionally, it is known that catalase activity is enhanced through phosphorilation by the c-abl tyrosine kinase (TK). In a preliminary work, our group found normal levels of GSH but high levels of antioxidant enzyme catalase in CML patients. The aim of this work is to evaluate the possible role of catalase as another pathway to Imatinib resistance. White blood cells from blood donors, CML patients and patients with reactional leukocytosis caused by infectious bacterian diseases were obtained using Ficoll-Hypaque. Catalase activity was measured by monitoring decomposition of 10mM H2O2 at 240nm according to the method described by Aebi. The viability of eritroleukemia cell lineage K562 previously treated or not with 100μM 3-aminotriazole (catalase inhibitor) was evaluated by MTT method with Imatinib 1, 5 and 25μM for 24 and 48 hours. Imatinib induced intracellular ROS generation was monitored by flow cytometry with 40μM 2′, 7′-dichlorofluorescein diacetate (DCFH-DA), a well-known ROS capturing reagent. Catalase activity was increased up to 30x in CML patients but not in reactional leukocytosis. When BCR-ABL positive cell line K562 was treated with Imatinib (50μM), catalase activity levels decreased in 38% compared with untreated cells. Also, we found that 3-aminotriazole can overcome the resistance to Imatinib in K562 cells resulting in 25% enhancement of dead cells. The protection of catalase against Imatinib-ROS generation was confirmed by DCFH-DA/FACs in cells incubated with PEG-catalase (monomethoxypolyethylene glycol covalent attached to catalase). Taken together, these results suggest that catalase might be another attractive target for overcoming resistance to Imatinib in Imatinib-resistant CML cells with high catalase activity.
A Prospective Analysis of Adverse Events in a Phase 2 Trial of Imatinib in the Treatment of Polycythemia Vera.
Authors: Jones C. Michael , Dickinson Tina M. 
Published: October 2019

Abstract
Imatinib has recently been demonstrated to be an effective treatment in Polycythemia Vera. Initial response rates are high, approaching 80%. Patients enrolled in our trial, Novartis CST 1571B (US 144), were asked to complete a questionnaire identifying side effects of imatinib known from previous trials in chronic myeloid leukemia and GIST. Questionnaires were completed weekly for the first month, biweekly for the second month, and monthly for the remainder of the study. Mean time on study is now 1.1 years. Target enrollment was 20 patients. Mean age was 54 years; male to female ratio was 1.3: 1. Fourteen patients were Caucasian and 2 African American. Twelve patients were naïve to treatment and 4 patients were previously treated with hydroxyurea. Two dose escalations were allowed sequentially to 600mg and 800mg daily based on response using the Polycythemia Vera Study Group Criteria. Toxicity was graded according to the NCI Common Toxicity Criteria, version 2.0. Patient reported toxicities are expressed as the total number of events reported during the 120 week study period. Diarrhea was the most common reported toxicity at 53 events. Periorbital edema (33), pruritis (39), fatigue (29), arthralgias (20), headache (18), nausea (19), reflux (38), and dry mouth (15) were the next most commonly reported events on study. Other toxicities reported less commonly were chest pain (3), depression (4), cough (5), insomnia (6), weight gain (3), taste changes (7), and depigmentation (3). Most toxicities were mild at grade 1 and resolved spontaneously within 30 to 40 weeks from the time of treatment initiation. The exceptions to this were diarrhea, periorbital edema, and reflux esophagitis, which persisted in a number of patients throughout the study. The severity of gastrointestinal side effects was dose dependent. Previous studies of imatinib induced toxicity in patients with chronic myeloid leukemia have reported cutaneous manifestations as the most common event. Our study demonstrates the toxicities of imatinib in patients with polycythemia vera are generally mild and self limited, although resolution may take up to 35 weeks. In contrast with studies of imatinib and chronic myeloid leukemia, gastrointestinal toxicity was more common and tended to persist, but did not result in patient withdrawal from study. These differences from our findings may be the result of several factors: the use of a patient reported adverse events scale, small trial size, or differences in drug metabolism in a related myeloproliferative disorder.
A Photodynamic Pathway to Apoptosis Induced by Hypericin in Human Myeloma ARH-77 Cell Line: Possible Relevance to Photodynamic Therapy.
Authors: Avcu Ferit , Ural Ali U. , Baran Yusuf , Sobaci Gungor , Deveci Mustafa , Cetin Turker , Pekel Aysel , Sarper Meral , Elci Pinar , Gunduz Ufuk 
Published: October 2019

Abstract
Hypericin, a photosensitizing plant pigment from Hypericum perforatum, is a protein tyrosine kinase inhibitor that has been exploited in models for anti-tumor and anti-viral activity. In this study, we investigated the effects of hypericin on the human myeloma cell line ARH-77 as a model to determine whether hypericin-induced cell death is available. The cells were incubated with hypericin at concentrations ranging from 0.001 to 10 microg/ml in RPMI at 37°C in 5% CO2 atmosphere for 4 h. Then, the cells were irradiated at 532 nm (fluence=24 J/cm2) using a dye laser pumped by an argon laser (Orion). After 72 h exposure, the IC50 of hypericin was 0.005 microg/ml as determined by the MTT assay. Hypericin exerted phototoxic effect on ARH-77 cells, while it did not produce toxic effect in the absence of irradiation. After 72 h exposure to 0.005 microg/ml photoactive hypericin, apoptosis was assessed by morphological changes, DNA fragmentation and flow cytometric analysis using Annexin V and propidium iodide staining. Most of the cells were accumulated in the late stage of apoptosis and these cells were brightly stained and fragmented nuclei and cytoplasmic blebbing were observed. A decrease in the number of apoptotic cells was detected when protein kinase C was inhibited by addition of staurosporine to photoactive hypericin induced ARH-77 cells. From these results, we demonstrated that exposing myeloma cell line ARH-77 to photoactive hypericin inhibits cell growth in a dose dependent manner, induces apoptotic cell death by protein kinase C activation, and provides a rationale for potential applications in vivo.
Normal Bone Marrow Stromal Cells Inhibit the Growth of Myeloma Cells While Myeloma Bone Marrow Stromal Cells Improve It.
Authors: Zhuang Junling , Wu Yongji , Wang Xuan , Sun Wanling 
Published: October 2019

Abstract
Objective: Observe the effect of normal and myeloma bone marrow stromal cells (BMSC) to the growth of myeloma cells. Evaluate the function of adhesion molecules in the mutual action of BMSC and myeloma cells.
Methods: Immunomagnetic beads and anti-CD138 antibody were used to separate myeloma cells from patients’ bone marrow of multiple myeloma (MM). U266 and freshly purified myeloma cells respectively co-cultured with normal BMSC and myeloma BMSC. Single U266 or fresh myeloma cells were cultured as control group. Anti-CD11a, anti-CD29, anti-CD44 and anti-CD49d antibodies were respectively added in the co-cultured system and single myeloma cells. After culture for 72 hours, cell cycle of myeloma cells and the ratio of apoptotic cells were tested by flowcytometry. The level of interleukin 6 (IL-6) in the supernate was measured.
Results: Normal BMSC inhibited the growth of myeloma cells while myeloma BMSC improved it. As to U266, S+G2% of myeloma cells in normal BMSC group and myeloma BMSC group were 55.5±4.0% and 65.4±4.5%(p<0.05)respectively. The ratio of apoptotic cells were 3.92±1.83% and 0.86±0.58%(p<0.05)respectively(figure1 from left to right: U266, U266+normal BMSC, U266+myeloma BMSC, red: apoptotic cells). The level of IL-6 in the supernate were 1236±146.2pg/ml and 1661±161.5pg/ml(p<0.05)respectively. After co-cultured with normal BMSC and myeloma BMSC, S+G2% of freshly separated myeloma cells were 5.2±4.1% and 15.9±5.5%(p<0.05)respectively. The ratio of apoptotic cells were 18.7±7.8% and 8±3.9%(p<0.05) respectively (figure2 from left to right: fresh myeloma cells, fresh myeloma cells+normal BMSC, fresh myeloma cells+myeloma BMSC, red: apoptotic cells). The level of IL-6 in the supernate were 1621.2±121.1pg/ml and 2151.1±170.4pg/ml(p<0.05)respectively. The function of four adhesion molecules was different in the co-culture system. Anti-CD11a hardly affected the growth of myeloma cells. But anti-CD29 and anti-CD44 showed their significant inhibition to the growth of myeloma cells, including the decrease of supernatant IL-6 level, percentage of phase S+G2 and the increase of apoptotic cells (fresh myeloma cells). Anti-CD49d indicated partial inhibition, but no significant change.
Conclusion: We firstly proved that normal BMSC inhibited the growth of myeloma cells. Anti-CD29 and anti-CD44 can block the adhesion of myeloma cells and BMSC, resulting in the growth inhibition of myeloma cells.
Figure Figure
Whole Body MRI Versus Skeletal Survey in Multple Myeloma.
Authors: McHugh Johnny , Johnston Ciaran , Duke Deirdre , Thornton Patrick , Eustace Steve , O’Gorman Peter 
Published: October 2019

Abstract
BACKGROUND: Bone involvement in myeloma is conventionally assessed by radiographic skeletal survey (plain x-rays of spine, skull, chest, pelvis and long bones). However this may not pick up bony involvement in all patients who may then present with serious complications of myeloma bone disease such as spinal cord compression. Whole body MRI may be better than skeletal survey at evaluating myeloma bone involvement.
AIMS: To compare the evaluation of myeloma bone involvement by conventional radiographic skeletal survey (RSS) with whole body magnetic resonance imaging (MRI).
METHODS: 35 patients with multiple myeloma (median age 68 yrs, range 46–81) underwent conventional RSS and whole body MRI. 19 of the patients had newly diagnosed multiple myeloma and 16 had relapsed multiple myeloma. The extent of myeloma bone involvement was evaluated in both RSS and MRI as follows: the body was divided up into ten areas: skull, cervical spine, ribs, thoracic spine, lumbar spine, pelvis, right arm, left arm, right leg, left leg. In each area the extent of myeloma bone involvement was scored in both RSS and MRI as follows: 0 = normal; 1 = one focus of abnormality; 2 = more than one focus of abnormality; 3 = diffuse disease. The scores for each of the ten areas were combined to give an overall score out of thirty for both RSS and MRI.
RESULTS: 30 of the 35 patients (85.7%) had evidence of bone involvement on MRI. This compares with 22 out of the 35 (62.9%) on RSS. The mean score for the extent of myeloma bone involvement on MRI was significantly higher than that for RSS (MRI mean score: 15.5 out of 30 (median 17, range 0–30); RSS mean score: 5.5 out of 30 (median 3, range 0 to 24); p<0.001). MRI was superior to RSS in all ten areas evaluated both in terms of lesion detection and extent of disease. The greatest difference between MRI and RSS was seen in the cervical, thoracic and lumbar spine, while the smallest difference was seen in the ribs and skull. Eight of the patients had no bone involvement detectable on RSS but did have bone involvement on MRI and this resulted in upstaging on Durie-Salmon staging in four patients.
SUMMARY/CONCLUSIONS: RSS has limited sensitivity and a significant ionising patient dose. It is a cumbersome procedure taking up to 30 minutes. Whole body MRI gives improved sensitivity and appreciation of anatomic location of disease. It is non-ionising and can be rapidly acquired at low cost. We conclude that whole body MRI is superior to conventional RSS in both the identification and evaluation of extent of bone involvement in multiple myeloma.
Establishment and Characterization of a New Human Myeloma Cell Line WuS1.
Authors: Sun Wanling , Wu Yongji , Li Hui , Wang Xuan , Zou Nong , Zhuang Junling 
Published: October 2019

Abstract
A new human myeloma cell line WuS1 was established from the bone marrow of a 45-year-old Chinese male patient with IgGλ type multiple myeloma (stage IIIB). The growth of WuS1 cells is constitutively independent of exogenous growth factors of feeder cells. The WuS1 cell line proliferated consistently as free-floating single cells in suspension, sometimes in small clusters or slightly adherent on the bottom of the plastic culture flask, without forming clumps. The cell line has been maintained without any external growth factors for over a year, and cells frozen in liquid nitrogen can be revived successfully. The doubling time of the cells was about 11 hours and the colony-forming rate was 55.56±6.33%. WuS1 displayed immature plasma cell features with an obvious heterogeneity in size and a high nuclear-cytoplasmic ratio in Wright-Giemsa staining.
Figure Figure
They were positive for ALP, CE, ACP(not inhibited by tartrate) and PAS stainings and negative for POX and NBE. By transmission electron microscopy, the cytoplasm of WuS1 contained abundant mitochondria, and parallel endoplasmic reticulum or Golgi apparatus in some cells. The monoclonal immunoglobulin G and λ light chain were positive in cell lysate and not in cell culture supernatants by immnuoelectrophoresis. The cell surface antigens were positive for CD3, CD59, CD106 and CD138, and negative for CD4, CD5, CD8, CD10, CD13, CD14, CD19, CD20, CD22, CD29, CD31, CD33, CD34, CD38, CD44, CD49d, CD45, CD54, CD56 and HLA-DR by flow cytometry. Chromosomal analysis revealed a hypodiploidy and complex karyotype. WuS1 cells were negative for Epstein-Barr virus by PCR using EBV nuclear antigen-1 specific primers. Twelve SCID mice were injected with WuS1 cells intravenously or subcutaneously, and obvious tumor infiltration in bone marrow, liver, spleen, lung, kidney and injection site (subcutaneously group) were observed by pathologic examination.
Figure Figure
The novel WuS1 cell line will be useful in the study of the biology, etiology and treatment of multiple myeloma.