bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cardiopulmonary Testing and Pulmonary Hypertension in Patients with Sickle Cell Anemia
Authors: Chapdelaine Hugo , Soulieres Denis , Poirier Claude 
Published: April 2020

Abstract
Objectives: Sickle cell disease (SCD) is responsible for multiple complications that can lead to dyspnea, including pulmonary hypertension (PAH). PAH is a common cause of death among these patients and represents a prognosis feature, but can be clinically difficult to detect. We reviewed the charts of patients with SCD and analyse correlation between hemoglobin level, EKG, chest X-ray, pulmonary function tests, and cardiopulmonary exercise test (CPET) with echocardiography.
Methods: We performed a retrospective chart review of 25 patients with SCD (11 patients with SC, 12 patients with SS and 2 patients with S-β thalassemia haplotype). All patients had pulmonary functions testing, CPET and resting echocardiography in the same period. PAH was defined as tricuspid regurgitant jet velocity (TRJV) &gt;2,5 m/s on cardiac echography. Subjects underwent symptom-limited CPET using cycle ergometry and individualized ramp protocol. Exercise was stopped when subject showed criteria of maximality or inability to keep the pace. Breath-by-breath analysis was done.
Results: Only 3 patients had PAH. All these patients were SS and suffered numerous acute thoracic syndrome. No correlation were made between lower hemoglobin level and higher TRJV; no patients had EKG or CXR anomaly. DLCO/VA appeared to be a independent risk factor: DLCO/VA &lt; 75 % correlated with a TRJV &gt; 2,5. CPET was abnormal in all patients, but were limited by patient fatigue and patterns were consistent with peripheral muscle disease except for one patient who had PAH; this patient showed a higher VE/VCO2 and a slower pulse kinetics, compatible with pulmonary vasculopathy.
Conclusions: Decrease DLCO/VA may indicate a sickle pulmonary vasculopathy. and this factor appears to play a significant role in detecting PAH in SCD patients. CPET was a safe procedure and gave details about global aerobic performance and exercise tolerance. Because of peripheral muscle limitations, CPET still an unreliable diagnostic test for PAH. Peripheral muscle dysfunction could explain a large part of exercise intolerance, but physiopathologic mechanisms still underexplored.
Differential Effect of the Histone Deacetylase Inhibitors Vorinostat and MGCD0103 in Cell-Free and Cell-Based Assays
Authors: Buglio Daniela , Khaskhely Noor M , Georgakis George V , Younes Anas 
Published: April 2020

Abstract
Recent clinical trials demonstrated a promising clinical activity of a variety of Histone deacetylase inhibitors (HDACi) in patients with relapsed non-Hodgkin and Hodgkin lymphoma (HL). In this study, we examined the in vitro activity of the pan- DAC inhibitor vorinostat and the isotype selective HDAC inhibitor MGCD0103. Using a cell free fluorogenic peptide-based substrate assay, both MGCD0103 and vorinostat were potent inhibitors of class I HDACs, with vorinostat also effectively inhibiting HDAC6. Subsequently, we examined the effect of these two compounds on three HL-derived cell lines (HD-LM2, L-428, and KM-H2). Although these cell lines had a similar expression level of class-I HDAC proteins, as determined by western blot analysis, they were more sensitive to MGCD0103 in short term culture. After 72 hours of incubation, the IC50 of MGCD0103 was consistently &lt;0.5 μM compared with &gt;1.5 μM for vorinostat. This differential antiproliferative effect was also observed at the molecular level. Submicromolar concentrations (as low as 0.1 μM) of MGCD0103 effectively induced the expression of p21 and the acetylation of H3 histones, whereas higher concentration of vorinostat were required to achieve a similar effect. Furthermore, we examined the effect of equimolar concentrations of MGCD0103 and vorinostat on cytokine and chemokine secretion, using a multiplex cytokine array (34 cytokines/chemokines), on a selected panel of gene expression using pathway PCR array, and on selected protein expression using pathway protein array (phospho kinases and apoptosis pathways) and multicolor FACS experiments. While vorinostat and MGCD0103 shared similar effects on a variety of these elements, MGCD0103 was more effective in upregulating p21 expression, activating the intrinsic caspase pathway, and favoring the secretion of Th1-type cytokines. Given the recent reported results from two independent phase-II studies in patients with relapsed HL that demonstrated higher response rate achieved with MGCD0103 compared with vorinostat, our results suggest that cell-based assays may have a better predictor value of the potential clinical activity of HDACi in HL compared with the cell-free assays.
Incidence and Prognostic Value of FLT-3 and NPM1 Genes Abnormalities in Acute Myeloid Leukemia in the Côte D’or Population, France
Authors: Maynadié Marc , Courtois Martine , Mounier Morgane , Janoray-Manivet Ines , Lafon Ingrid , Guy Julien , Carli Paule-Marie , Girodon François 
Published: April 2020

Abstract
Context: In acute myeloid leukemia (AML), the recently described FLT-3 and NPM1 genes abnormalities were found to have a prognostic value in AML with normal karyotype and a specific therapeutic strategy was proposed according to these abnormalities. We look for the incidence and prognostic value of these abnormalities in cases diagnosed on a well defined population.
Material and Methods: AML diagnosed according to WHO classification between 01/01/2001 and 31/12/2006 in the population of the Côte d’Or department, were included. Karyotype analyses were performed in 81% of the cases. The FLT3 D835 mutation, the FLT3 internal duplication (ITD) and the NPM1 mutation were systematically studied on the biological material of the diagnosis cryopreserved in the Ferdinand Cabanne Biobank of Burgundy, by PCR and DNA sequencing techniques. The vital status of the patients was update on 31/10/2007. The relative survival was calculated with the STATA (V9) software.
Results: 100 de novo AML and 47 secondary AML (sAML) were registered (72 females and 75 males). The world standardized incidence rate was 2.4 in men and 1.5 in women for de novo AML instead of what it was respectively of 1.1 and 0.6 in sAML. The urban predominance was present in both type of AML. The karyotype was normal in 38% (45/119) of cases (35% of de novo AML and 21% of sAML). It was abnormal in 62% of cases (74/119)(51% of de novo AML and 49% of sAML). Molecular analyses were performed in 78 de novo AML and in 24 sAML. FLT3 ITD was present in 19% (15/78) de novo AMl and in any sAML. The FLT3 D835 mutation was present in 6.5% of de novo AML and in 8% of sAML. NPM1 was mutated respectively in 26% and 4% of the cases. There was a significantly higher WBC count and proportion of blast cells in peripheral blood in FLT3 ITD cases. Overall and relative survivals of FLT3 ITD cases were decreased compared to FLT3 wild type cases. No difference according to NPM1 status was found.
Conclusions: These data confirm the bad prognostic value of FLT3 ITD status in AML observed in clinical series. Furthermore their particular interest lies in the fact that they are the first molecular data in AML produced on a population-based series indicating the feasibility of such epidemiological studies.
Clonal Profile Analysis of Leukemic Progenitors and Diagnosis Blast Cells in Pediatric B-Cell Precursor Acute Lymphoblastic Leukemia
Authors: Veltroni Marinella , Paganin Maddalena , Frasson Chiara , Fabbri Giulia , Marzollo Antonio , Seganfreddo Elena , Giarin Emanuela , Fortunato Elena , Aricó Maurizio , Basso Giuseppe 
Published: April 2020

Abstract
Recent studies suggest that the majority of malignant cells found in B-cell precursor acute lymphoblastic leukemia (BCP-ALL) arise from a rare population of leukemic progenitors. Little information is available on the presence of clonal rearrangements in cells at the stage of early precursor. To address this issue we analyzed clonality profile of early leukemic precursors sorted by flow-cytometry. Leukemic cells were obtained from bone marrow samples collected at diagnosis from 6 patients with childhood BCP-ALL. Furthermore, bone marrow cells were collected from 3 healthy children who were harvested for bone marrow donation. Three subpopulations of leukemic cells were investigated: total unsorted blasts, the sorted CD34+/CD38−/CD19+, and the sorted CD34+/CD38−/CD19− cells. Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements were screened by polymerase chain reaction (PCR) in the sorted populations and in the bulk leukemic cells in order to identify molecular markers of clonal evolution. Sequence analysis was then performed on the N-region. Overall, a total of 38 different Ig/TCR gene rearrangements were identified in the 3 cell populations under study (total blasts, CD34+/CD38−/CD19+, and CD34+/CD38−/CD19−). Of them, 13 (34%) were found in the three populations; 12 (31%) were found in two of the three populations: 7 in total blasts and CD19+ subset, 3 in total blasts and CD19−, 2 in CD19− and CD19+; finally, 13 were found only in one subpopulation: 4 in total blast cell, 5 in CD19+, 4 in CD19−. In all the six patients studied, BCP-ALL progenitors CD34+/CD38−/CD19− and CD19+ and the bulk tumor blasts shared at least one Ig/TCR gene clonal rearrangement with the same N-region. In 5 out of 6 patients at least one rearrangement detected in the BCP-ALL progenitors was undetectable in total blasts. Conversely, in 3 patients the clonal rearrangement observed in the bulk leukemic cells was not identified in any of the two sorted ALL precursor populations. Clonal rearrangement was never detected in the samples from healthy bone marrow donors. Our findings confirm that clonal rearrangement may be detected at the stage of early B-lineage precursor CD34+/CD38−/CD19−, suggesting that leukemic transformation may occur at this stage or even before in BCP-ALL. We plan to extend this observation by repopulating studies in NOD/SCID mice.
Down-Regulation of MicroRNA-143 and -145 in Childhood B-Lineage Acute Lymphoblastic Leukemia at Initial Diagnosis and in Relapse but up-Regulated When in Remission
Authors: Yang Yung-Li , Jou Shiann-Tarng , Lin Shu-Wha , Lin Dong-Tsamn , Hu Chung-Yi , Chang Sheng-Kai , Yen Ching-Tzu , Chiou Rong-Jing , Lin Kai-Hsin , Lu Meng-Yao , Chang Hsiu-Hao , Lin Shu-Rung 
Published: April 2020

Abstract
Inappropriate expression of microRNAs (miRNAs) is strongly associated with leukemogenesis. miRNAs (miRs)-143 and -145, previously shown to be reduced in colon cancers, have been demonstrated recently to be down-regulated in B-cell malignancies including chronic lymphoblastic leukemia, B-cell lymphoma, Burkitt’s lymphoma. In this study, we determined the role of miR-143 and miR-145 in childhood leukemia treated by the standard TPOG (Taiwan Pediatric Oncology Group)-2002 protocols approved for treatment of patients in Taiwan. Ten patients with childhood B-lineage ALL and four ALL leukemia cells lines including REH (CRL-8286TM), CCRF-SB (CCL-120TM), RS4 (CRL-1873TM), and SUP-B15 (CRL-1929) were measured for the miRNA expression using a TagMan quantitative RT-PCR method. Our results showed that miR-143 and miR-145 were both down-regulated 0.09 and 0.19 times individually (n= 13, p &lt;0.001 and p = 0.011) in mononuclear cells in bone marrow of newly-diagnosed and relapsed samples in comparison with the same cell types of remission samples (n=16). To examine possible suppressive functions of miR-143 and miR-145 with respect to cell growth, the REH cells were used for expression with precursor form and mature form miR-143 and miR-145, and subsequently measured for cell growth rate. Preliminary results showed that the two miRNAs did not alter the growth rate of the REH over-expressing either miRNA. Taken together, we have identified miR-143 and miR-145 as biomarkers that may differentiate malignant and normal B cells. miR-143 and miR-145 may contribute to leukemogenesis in childhood B-lineage ALL by new yet to be defined mechanisms.
Complete Loss of the RUNX1 Gene: A Review of the Implications for Leukemogenesis and Patient Management
Authors: Stephenson Christine F. , Valenzuela Martha , Iyer Uma , Richmond Lioudmila 
Published: April 2020

Abstract
The RUNX1 (AML1) gene is a transcription factor that regulates expression of genes involved in hematopoietic cell differentiation. It is a gene located on chromosome 21 at q22. Genetic alterations of RUNX1 whether through loss-of-function point mutations, translocation or amplification are known to impact myeloid differentiation and trigger leukemic transformation in particular with respect to myelodysplasia and acute myeloid leukemia. However, while there are many articles describing the impact of these types of RUNX1 genetic alterations, there is a paucity of information regarding loss of the entire RUNX1 gene. The case in this abstract highlights the significance of understanding loss of the RUNX1 gene. An 87 year old patient presented for evaluation for anemia and leukopenia. Flow cytometric evaluation revealed 26% myeloid blasts and confirmed a diagnosis of acute myeloid leukemia (AML). The cytogenetic findings demonstrated a translocation between chromosomes 17 and 21 −t(17;21)(q11.2;q22). The dilemma then was to determine if this was a variant of the traditional t(15;17) associated with acute promyelocytic leukemia or a variant of the t(8;21) associated with the M2 subtype of AML. FISH studies determined that there was no involvement of the RARA gene and no evidence of a RUNX1/ETO rearrangement. However, there was a complete loss of RUNX1 on the abnormal chromosome 21. Thus, what appeared to be a balanced translocation included a cryptic loss of RUNX1. While this may be an interesting case presentation the more pertinent question is what is the impact of the RUNX1 loss? This case prompted a review of the data on complete loss of the RUNX1 gene which while limited suggests that RUNX1 loss on its own is not leukemogenic. This poster presents the data and implication of complete loss of RUNX1, the role of this loss in leukemogenesis and patient management.
Erythropoietin and Cyclophosphamide Combined Treatment Additively Enhances Immunoglobulin Production in Mice
Authors: Lifshitz Lilach , Berger Assaf , Avneon Maayan , Mittelman Moshe , Neumann Drorit 
Published: April 2020

Abstract
Erythropoietin (EPO) is an important component in the treatment of cancer patients for improvement of cancer related anemia. EPO treatment for cancer related anemia is usually combined with chemotherapy. Cyclophosphamide (CP) is a known cytotoxic alkylating agent widely used in cancer chemotherapy. While at high doses it functions as an immunosuppressive agent, the anti-neoplastic activities of CP at low doses are attributed to enhancement of cellular and humoral immunity e.g. (Berd et al., Cancer Res; 1984). We have previously shown that EPO displays anti-neoplastic activities (Mittelman, 2001, 2004) and that EPO treatment is associated with enhancement of both the humoral and cellular immune responses (Prutchi-Sagiv 2006, Katz 2007). Here we focused on a murine model of DNP-KLH-injection, used to assess the humoral response in mice. Recently we demonstrated that administration of high doses of EPO (180U×3) to DNP-KLH-injected C57BL mice resulted in an increase in anti-DNP immunoglobulin G1 (IgG1) production. The present study was designed to examine the effect of combining low dose CP (12.5mg/kg ×2) used to achieve an anti-neoplastic activity, with a lower dose of recombinant human EPO (rHuEPO; 90U×3) on the humoral immune response of the DNP-KLH-injected mice, thus simulating the conditions of patient care. Hence, we compared anti-DNP Ig serum levels in DNP-KLH-injected C57BL mice that were treated with either EPO or CP alone, or the combination of CP and EPO (CP-EPO). CP treatment alone resulted in increased levels of serum anti-DNP IgG1 (O.D.(CP) = 0.38±0.06 vs O.D.(Non treated) = 0.18±0.064). In contrast, EPO treatment alone enhanced serum levels of IgG2 (O.D.(EPO) = 0.47±0.09 vs O.D.(Non treated) = 0.18±0.069). CP or EPO alone did not affect the total levels of anti-DNP total Ig (O.D.(EPO) = 0.37±0.07 vs O.D.(Non treated) = 0.28±0.04). Yet, the combined CP-EPO treatment resulted in increased levels of anti-DNP total Ig (O.D.(EPO+CP) = 0.48±0.05 vs O.D.(EPO or CP) = 0.37±0.04), maintaining the higher levels of IgG1 (O.D.(EPO+CP) = 0.38±0.06) and IgG2 (O.D.(EPO+CP) = 0.49±0.1). In summary, the combined CP-EPO treatment additively improved immunoglobulin production, compared to treatment with CP or EPO alone. We thus demonstrate that in context of chemotherapy treatment as usually administered in the clinic, EPO can enhance humoral immunity alongside its erythropoietic activities. Our findings emphasize the role of EPO as an immunomodulator, particularly when given as treatment in a combined therapeutic panel
Proteinase 3 Mediates Direct Inhibition of CD4 and CD8 T Lymphocyte Proliferation by Inducing Enzyme-Independent G0/G1 Cell Cycle Arrest
Authors: Yang Tian-Hui , Quintanilla Kathryn E , Cortez Amy M , Molldrem Jeffrey J. 
Published: April 2020

Abstract
Proteinase 3 (P3), a serine protease found in primary granules in granulocytes, is the target of T cell- and B cell-mediated autoimmunity in Wegener’s granulomatosis (WG) and of anti-leukemia immunity mediated by PR1 (VLQELNVTV)-specific cytotoxic T lymphocytes (PR1-CTL). Although aberrant P3 and neutrophil elasase (NE) expression in leukemia increases susceptibility to PR1-CTL-mediated killing, overexpression of P3 also induces apoptosis of the high affinity PR1-CTL leading to deletional tolerance and leukemia outgrowth. Because expression of P3 and NE in sera from leukemia patients is increased by 5-fold compared to healthy controls, we sought to determine whether such overexpression of P3 or NE impairs PR1-CTL immunity to leukemia by a direct affect on T lymphocytes. To study this, T cells from healthy donors were activated by anti-CD3 and anti-CD28 and exposed to increasing concentration of P3 or NE over one to five days, and the percentage of apoptotic cells and cell proliferation were determined by flow cytometry using PI, anti-Ki-67, and CFSE. P3, but not NE, induced dose-dependent apoptosis of up to 30% of T cells, and in the non-apoptotic cells, a 50% inhibition of CD4 and CD8 T cell proliferation at 1 μg/ml and 100% at 10 μg/ml compared to untreated cells. This effect was not enzyme-mediated since prior exposure of P3 to 56°C or co-incubation with the serine protease inhibitors Elafin and alpha-1 antitrypsin showed no affect on apoptosis or cell proliferation. P3 induced a cell cycle arrest at the G0/G1 interface, determined with PI and Ki-67 staining of healthy donor T cells that were exposed to P3 for up to 3 days. In contrast, at protein concentrations up to 25 μg/ml, NE showed no such inhibitory effect on apoptosis or cell proliferation. In addition to its role as a leukemia-associated antigen, P3 is also targeted by the cANCA antibody in patients with WG and the serum titer correlates with disease activity. Therefore, we hypothesized that the effect of P3 on T cell proliferation might also be affected by humoral immunity during circumstances of systemic autoimmunity. Co-incubation of P3 with a molar excess of cANCA reversed P3- mediated inhibition of both CD4 and CD8 T cells, consistent with a role of this antibody and the P3 target antigen in controlling T cell autoreactivity. Taken together, this data shows a new role for P3 in regulating T cell proliferation, which occurs only at high P3 concentration, similar to P3 in sera from leukemia patients, which is not enzymedependent. This supports a direct role for P3 in regulating both anti-leukemia immunity and autoimmunity. This data will need to be considered for effective immunotherapy targeting P3 in leukemia patients and these inhibitory effects also suggest a role for P3 in regulating autoimmunity at sites of inflammation, such as in patients with WG.
Mono-Chlorinated 3-Carboxy-7-Hydroxycoumarin: A Highly Fluorescent and Water-Soluble Violet Excitable Dye for Cell Analysis
Authors: Abrams Barny 
Published: April 2020

Abstract
In our search for new violet excitable dyes with improved photochemical properties, we examined several halogen substituted 7-hydroxycoumarins and found that chlorinated derivatives could be as bright as their fluorinated analogs. In particular, mono-chlorinated 3-carboxy-7-hydroxycoumarin (V450) has a quantum yield equal to 3-carboxy-6,8- difluoro-7-hydroxycoumarin (Pacific Blue™). Antibodies specific to various cell surface markers (CD3, CD4, CD45) were labeled with V450 and tested in single color flow cytometry assays. Our results show that conjugates of V450 are brighter than the same clones labeled with Pacific Blue™. Additional studies were performed in which V450-antibody conjugates were included in 8-color cocktails, along with FITC, PE, PerCP-Cy5.5, PE-Cy7, APC, and APC-H7 conjugates, aimed at subsetting lymphocytes and myeloid cells. These cocktails met all standards of performance for the particular applications for which they were designed, including stability, lack of interaction between components, and brightness. The new V450 fluorochrome proved to be compatible with both FACSLyse and PharmLyse processing of EDTA anti-coagulated whole blood, and showed no signs of adverse effects on unfixed cells.
Antitumour Effects of Exosomes in Combination with CTX and Poly I:C
Authors: Chang Chunkang , Guo Fang , Fan Huahua , Li Xiao 
Published: April 2020

Abstract
Exosomes are vesicles that are approximately 30–100 nm in diameter and are produced by different cell types. Exosomes derived from dendritic cells (DCs) expressing major histocompatibility complex class I/II molecules and T-cell costimulatory molecules are useful in antitumour therapy. Here we examined the antitumour effects of cyclophosphamide (CTX), polyinosinic-polycytidylic acid sodium salt (poly I:C) and exosomes. Although exosomes could be used to treat tumour, the therapy effect was unsatisfied. In order to enhance the therapy effect, we used Poly I:C and CTX. Poly I:C had been proved to enhance DC maturation. CTX could suppress Treg cells. Tumour growth and survival time were monitored. We have established that treatment with exosomes prolongs the survival time,. Treatment with the combination of CTX + exosomes + poly I:C not only suppressed tumour growth but also prolonged survival time. Unexpectedly, poly I:C could not suppress tumour growth and prolong survival time in the group treated with CTX + poly I:C. Because we did not study the group treated with poly I:C + exosomes, we could not determine whether poly I:C provides beneficial effects in this combination. We have established that exosomes can prolong survival time And therapy with exosomes in combination with CTX and poly I:C not only suppressed tumour growth but also prolonged survival time.