bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect of Soluble P-Selectin in Coagulation
Authors: Korakas Ingrid , Guillen Darlene , Martin Erika , Tranholm Mikael , Barnett Thomas , Li Chester 
Published: April 2020

Abstract
P-selectin, a member of the selectin family, is a vascular cell adhesion molecule. It is expressed and stored in alpha granules of platelets and in Weibel-Palade bodies of endothelial cells, and upon activation, P-selectin is translocated/transferred to the membrane surface. A key function of the P-selectin is to mediate leukocyte, lymphocyte and platelet interactions in inflammation and possibly in the thrombus formation. A soluble variant, S-Psel, comprising the extracellular domain of P-selectin, has been identified in healthy individuals, but is markedly elevated in patient with vascular disorders. Recent work on S-Psel suggested that S-Psel may play a role in hemostasis/coagulation through the generation of procoagulant TF-bearing microparticles (MP), and therefore, has potential in treating patients with the bleeding disorders, like hemophilia. The aim of this study is to verify studies reporting that S-Psel exhibits in vitro and in vivo pro-coagulant activity. S-Psel and S-Psel-Fc (IgG) fusion were purchased commercially or prepared at Novo Nordisk Research US (NNRUS) and their biological activity was verified by P-selectin/Pselectin glycoprotein ligand-1(PSGL-1) interaction in vitro. The clotting times of human whole blood and plasma treated with S-Psel or S-Psel-Fc, or with an irrelevant human IgG control protein, were measured by thromboelastography and aggregometry respectively. After up to 8 hours of incubation with S-Psel and S-Psel-Fc at a concentration of 15ug/ml, we found no significant difference between samples treated with S-Psel, S-Psel-Fc and the IgG controls. The ability of S-Psel to generate TF-bearing microparticles in human whole blood was examined in a FXa substrate cleavage assay; however, no significant difference in cleavage was observed. Finally, we evaluated S-Psel in vivo. Hemophilia A mice were injected with recombinant mouse S-Psel-IgG or S-Psel-Fc (IgG) at the concentration of 1.2 mg/Kg body weight and human IgG was used as control. As suggested from published results, the effect of S-Psel was determined 6 h after the treatment. Contrary to previous reports, the results revealed no significant difference in bleed time and blood loss between the experimental and control group. In conclusion, we were unable to demonstrate the procoagulant activity of S-Psel in our laboratory either in vitro or in vivo.
Anti-Thrombotic Effects of Targeting Outside-in αIIbβ3 Signaling in Platelets
Authors: Ablooglu Ararat J. , Kang Jian , Petrich Brian G. , Ginsberg Mark H. , Shattil Sanford J. 
Published: April 2020

Abstract
Platelet activation by agonists stimulates high affinity binding of adhesive ligands, such as fibrinogen, to integrin αIIbβ3. In turn, ligand binding initiates Src kinase-dependent outside-in signaling, platelet spreading on sub-endothelial matrices, and platelet aggregation. Short-term integrin blockade by parenteral αIIbβ3 antagonists is effectively anti-thrombotic in certain clinical situations, but long-term blockade by oral αIIbβ3 antagonists has been problematic. As an alternative approach, we evaluated the feasibility of disrupting outside-in αIIbβ3 signaling in vivo. A “β3(Δ760-762)” knock-in mouse was generated that lacked the three C-terminal β3 amino acid residues (RGT) necessary for αIIbβ3-dependent c-Src signaling, but retained β3 residues necessary for agonist- and talin-dependent fibrinogen binding. β3(Δ760-762) mice were compared with wild-type β3+/+ littermates, β3+/− heterozygotes, and “β3/β1(EGK)” knock-in mice where β3 RGT was replaced by three C-terminal residues in β1 (EGK) to potentially enable outside-in integrin signaling by certain Src kinases other than c-Src. Whereas β3+/+, β3+/− and β3/β1(EGK) platelets spread and underwent tyrosine phosphorylation normally on fibrinogen, β3(Δ760-762) platelets spread poorly and exhibited reduced tyrosine phosphorylation of c-Src substrates, including the β3 cytoplasmic domain itself (Tyr-747). Furthermore, unlike the three groups of control mice, all of the β3(Δ760-762) mice tested were protected from carotid artery occlusive thrombosis following vessel injury with FeCl3. The median tail bleeding time for the β3(Δ760-762) mice was approximately two-fold longer than that of the control mice. However, β3(Δ760-762) mice did not exhibit spontaneous bleeding, excess bleeding after surgical exposure of the carotid artery, fecal blood loss or anemia. Fibrinogen binding to β3(Δ760-762) platelets was normal in response to saturating concentrations of agonists to the PAR4 thrombin receptor or the GP VI collagen receptor; however, responses to ADP were impaired. These studies establish that deletion of β3 RGT disrupts outside-in αIIbβ3 signaling and confers protection from arterial thrombosis in vivo. Currently available anti-platelet drugs either inhibit agonist-induced activation of αIIbβ3 or block fibrinogen binding to the integrin. The studies reported here suggest that targeting outside-in αIIbβ3 signaling may represent an alternative anti-thrombotic strategy.
Identification and Functional Characterization of Novel Telomerase Variant Alleles in Patients with Bone-Marrow Failure Syndromes
Authors: Carroll Kathryn A , Calado Rodrigo , Young Neal S , Danzy Shamika , Ly Hinh 
Published: April 2020

Abstract
As multiple studies have shown a link between mutations in telomerase components and the disease pathogenesis of bone-marrow failure syndromes (BMFS), we screened blood or marrow cells from patients with various forms of blood disorders for novel pathogenic mutations in the telomerase hTERC and hTERT genes. We identified several heterozygous mutations in these genes, which were observed only in patient samples and not in controls. As these mutations are predicted to affect proper telomerase function, we introduced these and other mutations identified previously in patients with other forms of blood disorders into telomerase-negative cells in order to reconstitute telomerase enzymatic activity and to assess the effect of the individual mutations on modulating a wild-type telomerase function. All disease-associated mutations disrupted telomerase function to various degrees and most modulated a wildtype telomerase enzymatic function by haploinsufficiency based on results obtained from the telomeric repeat amplification protocol (TRAP). We also tested several previously reported natural sequence changes in the promoter region of the hTERC gene for their effect on gene expression and telomerase function via luciferase-reporter assay, TRAP, and Northern blot. The results obtained from these studies appear to contradict those that have recently been reported for some of the naturally occurring hTERC-promoter associated mutations. This study, therefore, offers new insights into the mechanism of natural telomerase mutations in regulating telomere lengths and marrow stem cell renewal capacity in patients with hematological disorders.
Possible Selection for the Jak2 V617F Mutation in Bone Marrow Failure
Authors: Dunsmore Sara , Williams Gaynor J. , Houston Donald S 
Published: April 2020

Abstract
The acquired JAK2 V617F mutation has been well documented to be common in the classical myeloproliferative disorders. As it is an activating mutation in a kinase involved in proliferative signaling in hematopoietic cells, it is presumed to play an etiopathogenic role. The same acquired JAK2 V617F mutation has been described in other hematological disorders such as myelodysplastic syndrome with a prevalence which, though much lower than in myeloproliferative disorders, exceeds what could be attributed to the chance coincidence of two rare disorders. The direction of causality in this association is unclear. CASE DESCRIPTION: A 42 year old woman initially presented at the age of six with anemia and was subsequently diagnosed with congenital dyserythropoietic anemia (CDA) type I. At initial presentation her other blood counts, including platelets, were normal. When she returned to hematology clinic as an adult, her anemia persisted, but she had elevated platelet and white blood counts. Bone marrow biopsy showed hypercellularity with increased representation of the myeloid and megakaryocyte lineages. It also showed megaloblastoid dyserythropoiesis and binucleate erythroid precursors. These changes are consistent with essential thrombocythemia superimposed on CDA type I. The marrow cells were heterozygous for the JAK2 V617F mutation.
DISCUSSION: This patient has a unique pairing of congenital dyserythropoietic anemia and a myeloproliferative disorder. No archival tissue is available so it is impossible to be certain she did not have the JAK2 V617F mutation on initial presentation, although the normal platelet and white blood count at age 6 support the inference that the myeloproliferative disorder arose later. Among patients with essential thrombocythemia, those who have the JAK2 V617F mutation tend to have higher hemoglobin levels, though in her case the development of the JAK2 V617F mutation did not lead to amelioration of her anemia. We hypothesize that the hematopoietic failure state associated with CDA may, like that of myelodysplasia, create an environment in which a genetically-mediated proliferative advantage due to an acquired mutation may be especially favored, and may thus have been a factor in the development of the myeloproliferative disorder.
Electronic Cell Volume and ALDH Expression of Hematopoietic Stem Cells Obtained by Apheresis
Authors: Shariatmadar Sherry , Sharma Siddharth , Cabana Raquel , Krishan Awtar 
Published: April 2020

Abstract
Small cell volume or diameter (3–7μm) appears to be one of the characteristics that may be used to identify early stem cells. Hematopoietic stem cells may be characterized by the expression of cell surface markers such as CD34, CD117 and CD133. In addition, stem cells such as “SP cells” can be recognized by the efflux of dyes such as Hoechst 33342 and presence of aldehyde dehydrogenase (ALDH). In prior studies we used the Beckman Coulter® Cell Lab Quanta(™) SC MPL analyzer to determine the electronic cell volume, diameter and marker expression (CD34, CD90, CD117 and CD133) of cells from peripheral blood apheresis (HPC, Apheresis) samples (Shariatmadar et al. Cytometry B, 74B:182–188, 2008; Sharma et al. Cytometry A, 73A:160–167, 2008). In the present study, we used this analyzer to determine cell volume, diameter and marker expression of ALDHpositive stem cells from HPC, Apheresis samples of 44 patients with hematologic malignancies mobilized with granulocyte colony-stimulating factor. The mean electronic volume of the ALDHpositive cells was 286μm3 (SD±27) corresponding to a diameter of 8.2μm. The mean percentage of ALDHpositive cells analyzed was 0.51% (SD± 0.004). CD34 expression was seen in 0.13% (SD±0.001) of the ALDHpositive cells. The mean electronic volume of the ALDH and CD34 positive cells was 270μm3 (SD± 33) corresponding to a diameter of 8.0μm. CD133, CD117 and CD90 expression was seen in 0.28% (SD± 0.01), 0.17 (SD± 0.05) and 0.04% (SD± 0.03) of the ALDHpositive cells. The mean electronic volume of the ALDHpositive cells with CD133 expression was 275μm3 (SD±29) corresponding to a diameter of 8.1μm while that of the ALDHpositive cells with CD117 and CD90 expression was 284μm3 (SD± 32) and 265μm3 (SD± 35), corresponding to a diameter of 8.15 and 7.9μm.
Stem Cell Marker Electronic Volume (μm3) Diameter (μm) CD90 (Thy-1) 300μm3 (SD± 38.5) 8.3μm (95% CI=8.1–8.4) CD117 (c-Kit) 349μm 3 (SD± 47.4) 8.7μm (95% CI=8.5–8.8) CD133 (Sca-1) 322μm3 (SD± 44.5) 8.5μm (95 % CI= 8.3–8.6) ALDH+ 286μm3 (SD± 27.0) 8.2μm (95 % CI= 8.1–8.3) ALDH+CD34+ 270μm3 (SD± 33.0) 8.0μm (95 % CI= 7.8–8.2) ALDH+ CD90+ 265μm3 (SD± 35.0) 7.9μm (95 % CI= 7.8–8.1) ALDH +CD117+ 284μm 3 (SD± 32.0) 8.15μm (95 % CI= 7.9–8.2) ALDH +CD133+ 275μm3 (SD± 29.0) 8.1μm (95 % CI= 8.0–8.3)
The use of electronic cell volume in conjunction with side scatter might provide a useful method for characterization of stem/progenitor cell populations with specific marker expression. In our study, hematopoietic stem cells expressing ALDH and bearing early stem cell markers were relatively larger than those previously reported. Future studies will aim to characterize the long term repopulating capability of this fraction of stem cells in in-vivo models.
Incidence of Febrile Episode During Stem Cell Mobilization (SCM) After High Dose Ciclophosphamide Chemotherapy (HD-CTX) and G-CSF (filgrastim or lenograstim) Administration in Multiple Myeloma (MM) Patients: II Interim Evaluation
Authors: Orciuolo Enrico , Buda Gabriele , Mauro E. , Marturano E. , Pastore Domenico , Cioppa Paola Della , Pietrantuono Giuseppe , Lombardini Letizia , Di Renzo Nicola , Irrera Giuseppe , Cangialosi Clotilde , Brunori Marino , Petrini Mario 
Published: April 2020

Abstract
Introduction: Clinical use of G-CSF in pts with high grade chemotherapy induced neutropenia does not conduce to a reduction of the incidence of febrile episodes (FE). This paradox may be explained by the acquisition of a defective chemotaxis by neutrophils (PMN) exposed to filgrastim (Fil), due to a higher adhesivity and cytoscheletric alterations. Lenograstim (Leno), a glicosilated form of G-CSF, is able to stimulate PMN production, manteining in vitro all the functional capabilities. On these bases, we hypotized that Leno may prevent FE and reduce their lasting in pts with chemotherapy derived neutropenia.
Patients and methods: starting from April 2005, 105 MM pts achieving HD-CTX for SCM were enrolled in 12 Centers. Treatment plan consisted in: HD-CTX (3 or 4 g/sqm) on day 1, G-CSF (random: Fil or Leno) 30 MU/day from day +4 to +9, 60 MU/day from day +10 to the achievement of an optimal CD34+ cell count for staminoapheresis. Random, 1:1, was effectuated on the base of a generated random list. FE, significant if equal or higher than 38 °C for at least 2 different determinations, were recorded till day +30. Primary endpoint is the incidence of FE; secondary endpoints are the duration of FE, efficacy in the CD34+ cell mobilization, time to mobilization.
Results: 105 pts were enrolled. All pts underwent post-chemo grade 4 neutropenia and G-CSF was administred starting from day +4. FE were recorded in 23 pts, 14 in the Fil arm (53 total pts) and 9 in the Leno arm (52 total pts). The global fever incidence was 21.9%, 26.4% with Fil and 17.3% with Leno, with a 9.1% difference. Average days with fever are 4.00 with Fil and 3.67 with Leno. Related to the neutropenia grade, 8 FE are recorded with Fil and 1 FE with Leno with absolute PMN count &gt;500/μL (grade 4); 7 episodes with Fil vs 1 with Leno when PMN are &gt;1000/μL (grade 3–4). CD34+ SCM occurs in after an average time of 10.3 day with Fil and 9.8 day with Leno, with an higher absolute count with Leno when compared to Fil: 131.9 CD34+/μL (range 40–640) vs 111.6 (range 40–616) CD34+/μL.
Conclusions: Leno is associated with a reduced incidence (17.3% vs 26.3%) of FE in MM patients undergoing to HD-CTX and SCM when compared to Fil. FE are recorded with Fil even in presence of PMN confirming the functional block by Fil on PMN documented in vitro. CD34+ mobilization occurs shorter and with higher efficiency with Leno when compared to Fil. On these evidences, patients’ enrollment will continue to 180 to validate these results.
Detection of Genetic Abnormalities for Diffuse Large B-Cell Lymphoma by Selective Interphase Analysis with FISH Method
Authors: Jiang Shi-Ping , Bouman Derek , Hovanes Karine 
Published: April 2020

Abstract
Diffuse Large B-cell Lymphoma (DLBCL) accounts for approximately 30% of all lymphoid malignancies. It can be difficult to separate DLBCL from Burkitt lymphoma and plasmablastic myeloma by morphology and flow cytometric immunophenotyping alone. Fluorescent in situ hybridization (FISH) targetgene analysis can assist the differential diagnosis by detection of genetic abnormalities associated with DLBCL.
Though no consistent numeric chromosomal abnormalities or translocations are observed as a hallmark for DLBCL, most cases have rearranged immunoglobulin heavy chain (IgH), BCL2 and MYC genes. Standard interphase FISH analysis is widely utilized to detect these genetic rearrangements. However, standard interphase FISH analysis could be falsely negative if the neoplastic cells are limited or if abundant benign cells are present in the background. Thus, the sensitivity to detect genetic abnormalities is low among the cases with a low number of neoplastic cells.
Herein we present three DLBCL cases in which interphase FISH analysis was applied on selective cells that are morphologically consistent with neoplastic large lymphocytes. All three cases showed genetic changes in a high percentage of the selected cells. These three cases were originally analyzed for IgH gene rearrangement, IgH/BCL2 and IgH/MYC translocation by using standard non-selected interphase study and showed false negative results. However, by review of the H&E slide and the flow cytometry results prompted a re-evaluation of the FISH studies using selective analysis of neoplastic cells only.
Our observations indicate that selective interphase FISH analysis improves the sensitivity of detecting genetic abnormalities in DLBCL. This analysis does require a corporation between hematopathologists and technologists performing interphase FISH studies.
Rel a Is a Novel Prognostic Marker in CLL That Is Independent of VH Gene Mutation Status, CD38 Expression and ZAP-70 Expression
Authors: Hewamana Saman , Lin Thet Thet , Rowntree Clare , Karunanithi Kamaraj , Burnett Alan , Brennan Paul , Fegan Chris , Pepper Chris 
Published: April 2020

Abstract
We recently demonstrated that the NF-kB subunit Rel A is associated with in vitro survival and clinical disease progression in CLL. We therefore hypothesized that Rel A would have prognostic significance in this disease. Rel A DNA binding was quantified in nuclear extracts derived from 131 unselected CLL patient samples using a quantitative DNA binding ELISA-based method. We tested the ability of Rel A to predict for the requirement for treatment and survival and compared our findings with other established prognostic markers. Rel A DNA binding was strongly associated with advanced Binet stage (P&lt;0.0001) but did not correlate with IgVH mutation status (P = 0.25), CD38 expression (P = 0.87) or ZAP-70 expression (P = 0.55). It was predictive of time to first treatment (P = 0.02) and time to subsequent treatment (P = 0.0001). In addition, Rel A was the most predictive marker of survival both from date of diagnosis (hazard ratio 9.1, P = 0.01) and date of entry into the study (hazard ratio 3.9, P = 0.05) and retained prognostic significance in multivariate analysis for both time to first treatment and overall survival in the presence of Binet stage, IgVH mutation status, CD38 and ZAP-70. Take together our data shows that Rel A is an independent prognostic marker of survival in CLL and appears to have the unique capacity to predict the duration of response to therapy. Prospective assessment of Rel A as a marker of clinical outcome and as a therapeutic target is now clearly warranted.
LPL and ADAM-29 Gene Expressions Stratify Prognosis in Clinical Stage BinetA and B Chronic Lymphocytic Leukemia Patients
Authors: Barreto Wolney Góis , Belluzzi Lorenza Oliveira , Siufi Grazziella Curado , de Andrade Silva Marçal Cavalcante , Kimura Eliza Yuriko Sugano , Vasconcelos Yuri , Arce Inara Lucia , Yamamoto Mihoko 
Published: April 2020

Abstract
Introduction: Chronic Lymphocytic Leukemia (CLL) is the most frequent adult leukemia in western countries. It is known to have a heterogeneous clinical course, some patients presenting an indolent disease while others have an aggressive disease requiring prompt treatment. Therefore, an individualized approach, especially in early clinical stage patients is necessary. Recent studies suggest that the biological markers LPL and ADAM-29 could be useful to predict prognosis in CLL patients: LPL being associated with an unfavorable prognosis while ADAM-29 to favorable one.
Aims: to evaluate the expression of LPL (L), ADAM-29 (A) and the L/A ratio in CLL patients in regard to Binet clinical stage and progression free survival (PFS).
Patients and Methods: thirty CLL patients followed at UNIFESP/EPM and HSPE were studied. RNA extraction was done by the TRIZOL method followed by c-DNA synthesis. c-DNA was amplified by PCR using LPL and ADAM-29 specific primers. T-Student’s exact test was used to compare the genes frequencies and Kaplan Meyer analysis to evaluate PFS. The results were considered statistically significant when p&lt;0.05.
Results: Of the 30 CLL patients studied (16M/14F, median age of 67 years, varying from 38 to 64 years) 19 (63,3%) were early clinical stage (Binet A), 8 (26,7%) Binet B and 3 (10%) Binet C. In the Binet A group LPL was negative in 52.63% of the patients and ADAM-29 positive in 63.15% with relation LPL-/ADAM29+ (or L/A=0) in 52.63% (10/19). In contrast among Binet B and C patients 90% (10/11) presented relation LPL+/ADAM-29- (or L/A=1). Those who were LPL+ or ADAM-29- or L/A=1 had a shorter PFS when compared to those LPL-or ADAM-29+ or L/A=0 (p&lt;0.05). Patients, even those in early clinical stages (Binet A and B), when LPL+, ADAM- and L/A=1 constitute a group with an unfavorable prognosis with a short DFPS (p&lt;0.05).
Conclusion: Our study shows that patients with a positive expression of LPL, negative for ADAM-29 and with L/A=1 have unfavorable prognosis independent of clinical stage. Analyzing Binet A and B together, we identified a subgroup LPL+, ADAM- and L/A=1 with a poor outcome. Thus, the expression profile of these genes could be useful to identify Binet A and B patients with an unfavorable prognosis, contributing to the decision making process to begin therapy in an individualized manner. (Support–FAPESP proc n. 05/57792-0)