bluuebunny/crossref_metadata_2025_split huggingface dataset:
ARRY-429520, a Novel KSP Inhibitor with Potent Anti-Proliferative Activity in Leukemic Cell Lines.
Authors: Lemieux Christine , DeWolf Walt , Voegtli Walt , Wallace Eli , Woessner Rich , Corrette Chris , Allen Shelley , Hans Jeremy , Zhao Qian , Aicher Tom , Lyssikatos Joe , Robinson John , Koch Kevin , Winkler James , Gross Stefan 
Published: April 2020

Abstract
Kinesins are eukaryotic microtubule-associated motor proteins. There are over 40 known kinesins, approximately 15 of which are closely associated with mitosis. Kinesin Spindle Protein (KSP), also known as Eg5, is a mitotic kinesin that is a required enzyme in mitosis (prophase / prometaphase). This protein plays a key role in the formation of the bipolar spindle, particularly related to its role in centrosome maturation/separation. Because KSP is expressed predominately in proliferating cells and is absent from postmitotic neurons, its inhibition should not produce the peripheral neuropathy associated with traditional microtubule disruption agents (taxanes and vinca alkaloids). We report here the in vitro characterization of a potent KSP inhibitor, ARRY-429520, a member of a series of KSP inhibitors discovered and optimized by structure-based design. ARRY-429520 inhibits human KSP, with an IC50 of 6 nM, by a mechanism which was demonstrated to be uncompetitive with respect to ATP and noncompetitive with respect to tubulin. It was shown to arrest cells in mitosis as measured by FACs analysis as well as the accumulation of phospho- histone H3, with an EC50 of 1.5 nM. Furthermore, this compound was demonstrated to be antiproliferative, with EC50s between 0.3 nM and 6.5 nM against a panel of human tumor cell lines, including various leukemia lines (K-562, KU-812, HL-60, KG-1, MOLT3, MOLT4). In addition, ARRY-429520’s potency in MDR-overexpressing cell lines was minimally impacted as compared to paclitaxel. Cellular imaging studies demonstrate that the normal mitotic spindle configuration was disrupted, with the formation of monopolar spindles, a hallmark feature of KSP inhibition, at single digit nanomolar concentrations of ARRY-429520. Markers of mitotic arrest and apoptosis were demonstrated in tumor xenografts from animals treated with ARRY-429520.
Sulforaphane Switches Cyclin E Isoform Expression and Blocks Leukemic Cell Cycling.
Authors: Gorham Laura M. , Frye Gigi , Miranda Michelle , Steinman Richard A. 
Published: April 2020

Abstract
Sulforaphane (SFN), an isothiocyanate derived from cruciferous vegetables, has been shown to inhibit the growth of prostate cancer cells in vitro and in vivo. We were interested in exploring potential antileukemic effects of SFN. The viability of multiple myeloid leukemic cell lines was decreased by 25uM SFN. Pharmacokinetic studies reported in rats suggest that this serum concentration can be achieved through oral dosing. Lower SFN concentrations (1–5 uM) inhibited leukemic cell growth without affecting cell viability. Synchronized HL-60 cells exposed to 25uM SFN were blocked at the G1/S phase transition. Kinetic analysis of cell cycle proteins demonstrated that the G1/S block arose from downmodulation of cyclins D3 and E rather than upregulation of cdk-inhibitors. Interestingly, we found that HL-60 cells expressed a low molecular weight (LMW, 36 kD) variant of cyclin E rather than (50 kD) full-length cyclin E. Treatment with SFN for as little as 2 hours caused a decrease in expression of the LMW cyclin E and induced the expression of a higher molecular weight (~50 kD) cyclin E isoform. Because LMW cyclin E has been associated with increased cdk2 activity and p27 resistance compared to full-length cyclin E, we postulate that SFN-mediated cyclin E isoform-switching contributed to growth inhibition of these leukemic cells. The signaling pathway through which SFN altered cyclin E expression appeared to be distinct from MEK/ERK and JNK pathways that have been implicated in the apoptotic effects of SFN. Given that cyclin E overexpression and, particularly, LMW cyclin E expression are correlated with poor prognosis in multiple cancers, the mechanism through which SFN decreases LMW cyclin E expression in these leukemic cells could have therapeutic significance.
Experimental Study on DNA Molecular Structure of Acute Myeloid Leukemia and Myelodysplastic Syndrome by FT-IR Spectra.
Authors: Wang Shihong , Xiao Zhijian , Han Ming-Zhe , Wu Qiang 
Published: April 2020

Abstract
So for, there were no definite molecular markers in most acute myeloid leukemia(AML) and myelodysplastic syndrome(MDS), and their molecular mechanisms have not been clarified. Their clinical diagnosis were mainly based on morphology of bone marrow and peripheral blood cells. The current study aimed to explore the mechanisms of abnormal DNA molecular structure in AML and MDS by Fourier Transform Infrared(FT-IR) spectra, and to explore the methods of early diagnosis and prognosis. At first, we segregated the mononuclear cells of AML-M2a, MDS and control groups samples, extracted the DNA of these cells, then we checked the completely dry DNA by FT-IR spectra and analysed the identity and difference among AML-M2a, MDS and Control groups. Comparing the mean DNA spectra of the AML-M2a, MDS and control groups revealed statistically significant difference(p&lt;0.001). Each group has an almost concordant FT-IR spectra rule. Compared with the control group, AML-M2a and MDS both were significant different(p&lt;0.001). Absorbance increasing of DNA in AML-M2a, MDS at the wavenumber 2880cm-1 and 2940cm-1 shown hypermethylation of nucleotide base(p&lt;0.001). Absorbance decreasing at wavenumber 1420cm-1 demonstrated deacetylation(p&lt;0.001). Absorbance decreasing at wavenumber 1150cm-1 indicated the abnormality of C—O,S=O,C—OH,C—C,C—O—C,C—N, and the absorbance increasing at wavenumber 1660–1590cm-1 reflected in-plane ring, C=N stretching vibrations and hypermethylation of cytosine. There were also other mild difference among AML-M2a, MDS and control groups. In summary, The principal chain, phosphonic backbone and nucleotide base of DNA in AML-M2a and MDS have significant difference with the control and could be the initial changes of DNA in AML and MDS occurrence and development. Based on the rules of change, FT-IR spectra is a potential method of early diagnosis and prognosis in AML and MDS.
Immunophenotypic Diagnosis of Lymphoproliferative Disorders Using the Abbott CELL-DYN Sapphire Hematology Analyzer.
Authors: Settegrana Catherine , Maloum Karim , Souni Fadila , Septier Isabelle , Vinet Sandrine , Bonnemye Patrick , Merle-Beral Helene 
Published: April 2020

Abstract
The CELL-DYN Sapphire (Abbott) is a routine hematology analyzer which uses integrated optical and fluorescence measurements, with FL1 (FITC) and FL2 (PE) detectors being configured for fluorescence analysis. Enumeration of CD3/CD4 and CD8 lymphocytes as well as platelets quantification using an anti-CD61 monoclonal antibody (moAb) are now included in the automated process of the analyzer. Recently, Johannessen et al. (Clin.Lab.Haematol.,2006) demonstrated that the CELL-DYN Sapphire analyzer allows a possible complete automated leukocyte immunodifferential count, using a panel of different moAbs (Johannessen et al., 2006).
Aim To test the ability of this blood analyzer to rapidly detect and quantify abnormal lymphoid cells by using a restricted panel of moAbs.
Patients and Methods Seventy blood samples were analyzed by using the following moAb combinations: CD3FITC/CD4PE, CD3FITC/CD8PE in an automated assay and CD3FITC/CD19PE and CD5FITC/CD19PE by using a manual procedure which only consists of mixing EDTA blood and antibodies. Results were obtained in 4 and 15 minutes for the automated assay and for the manual procedure, respectively. The results were compared with an extensive immunophenotype performed by using the FacsCanto cytometer (Becton Dickinson).
Results Sixty out of 70 samples were evaluated:
the quantification of CD3/CD4 and CD3/CD8 lymphocytes was well correlated with classical flow cytometry (FC), as previously demonstrated (R2 = .94 for CD3, .93 for CD3/CD4 and .95 for CD3/CD8). Interestingly, two T cell lymphomas (one with double positive T cells and one double negative T cells) were clearly recognized by the CELL DYN Sapphire and confirmed by classical FC and molecular biology. B-cells immunophenotyping: 25 samples with B cell lymphoproliferative disorders and with circulating abnormal lymphocytes (20 CLL, 1 HCL, 1 B-PLL and 3 MZL) were studied. In all cases, a very good correlation between the Abbott analyzer and classical FC was observed: CD19+/CD5+ (R2 = .99), CD19 (R2 = .91), CD3 (R2 = .91). Twenty controls without abnormal circulating B lymphocytes were studied and results were similar to those obtained with classical FC (R2 = .92 for CD3, .94 for CD19 and CD5). Of note in 10/70 samples, immunophenotyping using the Abbott analyzer was not possible because of the interference of lytic resistant red blood cells and/or platelets aggregates.
Conclusion The CELL-DYN Sapphire Abbott hematology analyzer provides a simple and fast immunophenotyping method useful for a preliminary screening in routine laboratory practice to detect and quantify abnormal circulating lymphoid cells simultaneously with the blood cell count and leukocyte differential.
Analysis of In Vitro Effects of Bortezomib (Velcade) on T Cell Prolymphocytic Leukemia (T-PLL).
Authors: Ozpuyan Fulya , Meyer Paul N. , Al-Masri Hytham , Ni Hongyu , Alkan Serhan 
Published: April 2020

Abstract
T-cell prolymphocytic leukemia (T-PLL) is an aggressive lymphoproliferative disorder with postthymic T cell phenotype and prolymphocytic morphology. In the majority of patients, the leukemic process progresses rapidly and patients die shortly after diagnosis (median survival of 7 months). Bortezomib, the first proteasome inhibitor to be approved for use in haematological malignancies such as multiple myeloma, is beginning to be utilized as an effective anti-neoplastic agent in other hematopoietic and non-hematopoietic neoplastic disorders. We report here the in vitro apoptotic effects of bortezomib on leukemic cells isolated from three T-PLL patients. Interestingly, one of the patient’s leukemia developed in the setting of immunosupression due to transplant therapy (post-transplant lymphoproliferative disorder). Flow cytometric analysis of leukemic cells of the three patients showed CD8, double CD4+CD8+ and double CD4−CD8− immunophenotypic features. All cases showed monoclonal band pattern by T-cell receptor (TCR) gene rearrangement as analyzed by the PCR amplification of the TCR gamma heavy chain gene. Freshly isolated leukemic cells with the CD8 phenotype T-PLL analyzed for apoptosis after ficoll hypaque separation and cultured in the presence of various concentration of Bortezomib (0.001, 0.01, 0.1, 1, and 10 uM) for dose curve analysis. Apoptosis of the leukemic cells was determined by Annexin-V and 7-AAD staining and flow cytometric analysis after incubation at 24 and 72 hours, respectively. Samples treated for 72 hours showed higher rate of apoptosis compared to 24 hours: 10 uM (62% increase above the base line of control cells), 1 uM (58%), 0.1 uM (55%), 0.01 uM (40%) and 0.001 uM (0%) concentrations while samples treated for 24 hours with 10 uM showed (42% increase above the base line of control cells) and 1 uM (33% increase above the base line). Light microscopic analysis of leukemic cells treated with Bortezomib confirmed that the majority of cells undergo apoptosis with Bortezomib treatment as it revealed nuclear fragmentation and apoptotic bodies. Leukemic cells recovered from cryopreservation from the second and third T-PLL patient samples analyzed also showed significant increase in early and late apoptosis at 24 hours with Bortezomib treatment (10nm). These results suggest that Bortezomib may provide an alternate therapy in the treatment of T-PLL. Future collaborative efforts investigating efficacy with Bortezomib as a single agent or in combination with other therapeutic agents will be crucial to improving survival for patients with this disease.
Clinical Significance of Alterations of the NOTCH1, FLT3 and p53 Genes in Pediatric T-Cell Acute Lymphoblastic Leukemia.
Authors: Park Myoung-ja , Taki Tomohiko , Shimada Akira , Sotomatsu Manabu , Hanada Ryoji , Hayashi Yasuhide 
Published: April 2020

Abstract
Activating mutations of the NOTCH1 gene have been reported in about 50% of T cell acute lymphoblastic leukemia (T-ALL). We performed mutation analysis of the NOTCH1, FLT3 and p53 genes by polymerase chain reaction followed by direct sequence. Mutations of the NOTCH1 were identified in 24 (30%) of 80 fresh samples and 10 (71.4%) of 14 T-ALL cell lines. Six missense mutations and 2 insertion in HD domain, 2 nonsense mutations and 6 insertions in PEST domains were found in 14 cell lines. Eight missense mutations, 9 insertions and one deletion in HD domains, 5 missense mutations, 3 nonsense mutations and 3 deletions in PEST domain were found in 80 fresh samples. The incidence of the NOTCH1 mutations is less frequent than that of previous reports. We observed about 95% of single nucleotide polymorphisms (5097 C/T) in HD domain, which is more frequent than 30% of previous report, possibly due to the racial difference. FLT3 internal tandem duplication, which is known as the poor prognostic factor in acute myeloid leukemia, were not identified in any T-ALL cell lines or fresh samples. Mutations of the p53 gene were found in 5 of 8 cell lines and 5 of 50 fresh samples. Mutations of both NOTCH1 and p53 genes were identified 3 of 8 cell lines and one of 50 fresh samples. In our study NOTCH1 mutations were not significantly associated with high WBC count and prognosis. Further studies are needed to find the association between novel genes and clinical features of pediatric T-ALL.
JNK/c-Jun Is Required for HMC-1 Cell Proliferation.
Authors: Wang Bin , Tsukada Junichi , Higashi Takehiro , Mizobe Takamitsu , Matsuura Ai , Mouri Fumihiko , Sawamukai Norifumi , Ra Chisei , Tanaka Yoshiya 
Published: April 2020

Abstract
Activation of c-jun N-terminal kinase (JNK) through c-kit-mediated phosphatidylinositol 3 (PI3) and Src kinase pathways plays an important role in cell proliferation and survival in mast cells. Gain-of-function mutations in c-kit are found in several human neoplasms. Constitutive activation of c-kit has been observed in human mastocytosis, acute myeloid leukemia, lymphoma, germ tumor and gastrointestinal stromal tumor. In the present study, we demonstrate that an anthrapyrazole SP600125, a reversible ATP-competitive inhibitor of JNK inhibits proliferation of human HMC-1 mast cells expressing constitutively activated c-kit mutant. We found that JNK/c-Jun was constitutively activated in HMC-1 cells without stimulation. When spontaneous activation of JNK/c-Jun was inhibited by treatment with SP600125, cell proliferation was suppressed. The concentration which effectively inhibited JNK/c-Jun activity in our experiment had no effect on SCF-induced phosphorylation of Akt or Erk, suggesting that SP600125 specifically inhibited JNK/c-Jun activity in HMC-1 cells. Moreover, we demonstrated that SP600125 induced HMC-1 cell apoptosis in dose- and time-dependent manner. Caspase-3 and PARP were cleaved as early as 12 h after treatment with SP600125, but caspase-9 was not. Also, cell cycle arrest in G1 phase was observed in SP600125 treated cells. Thus, the inhibitory effect of SP600125 on cell proliferation was associated with cell cycle arrest at the G1 phase and apoptosis accompanied by cleavage of caspase-3 and PARP. Caspase-3 inhibitor Z-DEVD-FMK almost completely inhibited SP600125-induced apoptosis of HMC-1 cells. In contrast, caspase-9 inhibitor Z-LEHD-FMK failed to block SP600125-induced apoptosis, suggesting that apoptosis induced by SP600125 was caspase-3 dependent. Following SP600125 treatment, down-regulation of cyclin D3 protein expression, but not p53 was also observed. Take together, JNK/c-Jun is essential for proliferation and survival of HMC-1 cells. The results obtained from the present study suggest the possibility that JNK/c-Jun may be a therapeutic target in diseases associated with c-kit mutant.
Late Appearance of t(9;22) at Relapse, in an Adolescent with an Homozygous 9p Deletion T-ALL.
Authors: Papadakis Vassilios , Papadhimitriou Stefanos , Paisiou Anna , Belesi Chrysoula , Parcharidou Agapi , Tourkantoni Natalia , Sambani Constantina , Paterakis George , Polychronopoulou Sophia 
Published: April 2020

Abstract
T-ALL has increasing incidence during adolescence and it is rarely associated with Philadelphia chromosome positivity. Moreover, extremely rare is the event of Philadelphia chromosome negativity at diagnosis and leukemia clone evolution with t(9;22) at relapse. Such a patient is being described and relevant issues are being raised. A 14 year old boy was diagnosed with leukemia due to malaise and ecchymoses. CBC revealed: WBC 170 k/uL (80% blasts), Hb 13.1 gr/dL, Ht 37.7% and PLT of 26 k/uL. Bone marrow confirmed the diagnosis of ALL (L2 morphology). Bone marrow flow cytometry depicted a T II ALL by EGIL classification (Tdt 54%, cCD3 99%, CD7 96%, CD2 96%, CD4 0%, CD8 2%, CD5 75%, TCRαβ 0%, CD1α 0%, sCD3 31%). Karyotype with G-banding, technically failed. By PCR, blasts were negative for TEL-AML1, E2A-PBXand MLL-AF4 while RNA coding for proteins p190 and p210 was also negative. Additionally, the 9 chromosome was evaluated by FISH technique (centromere and 9p21 areas/genes p16/p14 and p15- Vysis) and an homozygous deletion of 9p16 area was detected in the great majority of the blasts tested. The patient was treated according to the ALL-BFM-95 protocol and proved to be a poor prednisone responder, with residual disease detected on day +15 bone marrow and remission on day +33. He proceeded to the High Risk arm of the protocol. Thirteen months from diagnosis and 5 months on maintenance treatment, the patient suffered a bone marrow relapse. CBC revealed: WBC 30 k/uL (25% blasts), Hb 13.8 gr/dL, Ht 40.9 % and PLT of 201 k/uL. Bone marrow confirmed the relapse (L2 blast morphology) with a more mature T-cell type (cCD/TdT coexpression, with CD3 87% and TCRαβ 53.3% expression). Five of 25 metaphases revealed the following complex karyotype: 46,XY,del(6)(q21),?del(7)(q36), -9, t(9;22)(q34;q11),+mar1. By FISH , monosomy of 9 chromosome was not proved , while homozygous deletion of 9p16 area was again documented and t(9;22) was clearly evident. By PCR, there was no detection of RNA coding for proteins p190 and p210 with standard commercially available probes. The patient is treated with a fludarabine based regimen and imatinib, with the prospective of stem cell transplantation.
In conclusion, at diagnosis, T-ALL without t(9;22) but with an homozygous deletion of chromosome 9, was documented. At relapse, there is clone evolution with
a more mature T-origin blast detection of a complex karyotype with t(9;22). Ph+ T-ALL is rare, as well as rare is the event of ALL clone evolution, with emergence of Ph+ at relapse.
This patient’s data are being completed and there is prospective of further illuminating the underlying pathogenesis. Chromosome 9 deletions are under investigation for contributing in genetic instability. Homozygous 9p16 deletion of ALL blasts at diagnosis and relapse might play a key role in these events.
Changes in Thymic Recent Output Function in Patients with B-Cell Lymphocytic Malignancy.
Authors: Li Yangqiu , Yang Lijian , Chen Shaohua , Geng Suxia , Przybylski Grzegorz , Schmidt Christian A. 
Published: April 2020

Abstract
Since 1998 the use of signal joint T cell receptor excision circles (sjTRECs) to study changes in the frequency of recent thymic emigrants was reported, this study drew attention to the fact that the level of sjTRECs can be used to estimate the recent thymic output function. Therefore, sjTRECs was used as a new marker for analysis of thymic output function in different immunodeficiency diseases, immune reconstitution in patients after stem cell transplantation. Defects of cellular immunity, however, may also play a role in hematologic malignancies. Little is known about the feature of T-cell immune state in B-cell lymphocytic malignancy. In order to identify number of naïve T-cells in B-cell lymphocytic malignancy, sjTRECs-content was determined in the mononuclear cell fraction and the sjTRECs-number was related to the number of T-cells (according to the number of CD3-positive cells). Quantitative analysis of sjTRECs in DNA of peripheral blood T cells from 45 cases with B-cell lymphocytic malignancy (including 17 cases with ALL, 4 cases with CLL, 15 cases with B-NHL and 6 cases with MM) were preformed by real-time PCR and TaqMan analysis, and 14 normal individuals as controls. The results showed a dramatic reduction of sjTRECs values in patients. Some cases no sjTRECs copies could be detected in 40000 T cells. The mean value of sjTRECs was 0.39±0.75 copies/1000 PBMCs in ALL, 0.11±0.15 copies/1000 PBMCs in B-CLL, 0.67±1.39 copies/1000 PBMCs in B-NHL, 0.91±1.16 copies/1000 PBMCs in MM patients, as compared with 4.1±3.65 copies/1000 PBMCs in normal individuals, the sjTRECs level in all goups of B-cells lymphocytic malignancy were significant decrease (p=0.0001, p=0.048, p=0.002) except MM group (p=0.053). However, when comparison of the sjTRECs value in CD3+ compartments, it was not statistically different from the sjTRECs counts between patients with ALL (3.91±7.20 copies/1000 CD3+cells) and normal individuals (6.36±5.28 copies/1000 CD3+cells) (p=0.2139), suggesting that the reduction of sjTRECs level in patients wit ALL may due to lower numbers of T cells in peripheral blood. In conclusions, the results suggest that some thymic dysfunction in T cell generation was detected at least in most patient with B-cell lymphocytic malignancy. However, whether this is due to clonal expansion of T-cells to antigens, or reflects impairment of immune function associated to the malignancy, remains an open question. In a prospective study sjTRECs levels will be determined in different T- cell compartments including CD45RA+, CD4+ and CD8+ cells and so on.