bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of Multiphoton Intravital Imaging and a Multiscale Computational Model of Thrombus Development to Study the Role of FVII in Thrombogenesis
Authors: Kamocka Malgorzata , Xu Zhiliang , Chen Nan , Alber Mark , Rosen Elliot D. 
Published: April 2020

Abstract
Using 2-photon intravital microscopy we have generated high resolution, near real-time 3-dimensional images of a developing thrombus. Following Titanium - Sapphire laser-induced injuries in mouse mesenteric vessels, the developing thrombus was monitored by collecting stacks of confocal images through the developing thrombus. Data were collected in 3 channels for fluorescently labeled platelets, fibrinogen and 70,000 MW dextran. By including fluorescently labeled dextran in the blood we were able to monitor flow (plasma) and not labeled cells (leukocytes and erythrocytes) forming black silhouettes in the plasma. Since each 3-D reconstruction involves a series of scans, we were able to generate approximately 2–3 reconstructions per minute. Thus, the system sacrifices temporal resolution for high resolution structural information revealing the changing, heterogeneous sub-domain structure of the developing thrombus. The imaging system has been used to study the consequences of FVII-deficiency. Following injury, of the luminal surface of the vessel, a thin layer of platelets and fibrin accumulated at the injury site. Unlike injuries in wild-type mice where the thrombus continues to grow, the injuries in FVII deficient mice failed to grow as a result of frequent embolization from the developing structure. Interestingly, in a model of ferric chloride induced injury of the carotid artery, thrombi in FVII deficient mice form large structures capable of reducing flow although, unlike wild type mice, the FVII deficient animals fail to form stable occlusive clots. In parallel with the modified experimental vascular injury model, we have begun development of a computational model of thrombus development. The modeling framework consists of a stochastic and discrete Cellular Potts Model (CPM) to describe platelet and cellular interactions and continuous submodels to describe hydrodynamic and biochemical reactions. Our multiscale model includes the vessel wall, platelets (in resting and activated states), blood cells, coagulation reactions, fibrin formation, and hydrodynamic parameters as components. By comparing the in vivo experimental results with those of simulations varying the concentration of FVII we are able to refine and validate the computational model of thrombogenesis.
Synergistic Anti-Leukemic Effect of Combination Therapy with Anti-FLT3 Antibody IMC-EB10 and Methotrexate
Authors: Surguladze David , Corcoran Erik , Tonra James , Li Yiwen 
Published: April 2020

Abstract
FLT3 is a class III receptor tyrosine kinase overexpressed by blast cells in approximately 90% of acute myeloid leukemia and acute lymphoid leukemia (ALL) patients. Recent findings testing a small molecule FLT3 inhibitor (PKC412) in combination with eight conventional antileukemic agents, reported that methotrexate in particular did not combine well with PKC412 against leukemia cells (Furukawa, Y. et al., Leukemia 2007, 21: 1005–1014). IMC-EB10 is a fully human neutralizing antibody that binds FLT3 with high affinity (Kd 158 pM) and blocks FLT3 ligand binding to FLT3 (IC50 ~10 nM), leading to inhibition of MAPK, STAT5, and PI3K/Akt signaling in leukemia cells. Here we test for benefits of adding methotrexate to IMC-EB10. In an SEMK2 model of ALL we evaluated survival effects of treatments administered intraperitoneally, beginning 1 day after after intravenous injection of 5 × 10e5 cells/mouse. In this model the maximum tolerated dose (MTD) of methotrexate administered once per week for 3 weeks, was determined to be 100 mg/kg. At this MTD, Methotrexate increased median survival to 44 days, compared to 30 days in the control group (p&lt;0.0001 versus control by Log Rank Test). IMC-EB10 at its maximally efficacious dose (10 mg/kg, ip, 2×/week) increased survival in the SEMK2 model to 60 days (p&lt;0.0001 versus control). When these two regimens were combined, survival significantly increased to 83 days (p&lt;0.0001 versus all other treatment groups). Furthermore, 4 of 12 combination treated mice survived to 168 days after SEMK2 cell injection, but none of the mice in the other groups survived to this point. In this regard these agents were able to achieve an effect in combination that they could not achieve as monotherapies. These data support the significant therapeutic potential of this combination strategy in the treatment of leukemia.
Identification of Idiotype TCR Vα6, Vα10 and Vβ23 Relate to APL-Associated Antigen
Authors: Chen Shao hua , Li Yangqiu , Yang Li jian , Zhou Yu bing , Li Bo 
Published: April 2020

Abstract
Differentiation therapy with all-trans retinoic acid in combination with chemotherapy has significantly improved survival in patient with acute promyelocytic leukaemia (APL). However, relapse remains a major problem. In recent years, specific adoptive immunotherapy with tumor-specific T-cells was considered as a new approach to malignancy therapy. Recently, researches are focus on searching the generation of effective tumor antigen specific T-cells. The tumor antigen specific T cells could be determined by analysis of the T-cell receptor (TCR) Vα or Vβ repertoire, which display monoclonal or oligoclonal pattern. In order to develop the specific cellular immunotherapy in APL, in this present study, we analysis the clonality and the pattern of CDR3 sequence of TCR Vα and Vβ repertoire in peripheral blood T cells by RT-PCR and genescan technique, which was expected to define the APL associated TCR Vα or Vβ subfamily T cells. Oligoclonal T cells expressed TCR Vα6, Vα10 or Vβ23 genes were identified from one APL patient. The molecular characteristics of the CDR3 of Vα and Vβ genes rearrangement were Vα6-N-Jα17, Vα10-N-Jα35 and Vβ23-N-Dβ1-NJβ2.7. And the amino acid sequence of CDR3 region in TCR Vα6, Vα10 and Vβ23 subfamilies were CAMRENSAGNK, YLCAGDELLWECA or CASSSKWAGGTYEQY, respectively. These sequences were accepted by GenBank (Accession Number EU544946, EU544947 and EU647219). In conclusions, three idiotype Vα and Vβ sequences were identified in a case with APL, which was thought that may mediate the host specific immunity response for leukemia cells in APL. Farther investigation will be performed on construction the vector containing the antigen specific TCRs for gene transfer to establish TCR modified-T cells for specific immunotherapy of APL.
Feasibility of Inpatient Geriatric Assessment for Older Adults with Newly Diagnosed Acute Myelogenous Leukemia
Authors: Klepin Heidi D , Ellis Leslie Renee , Levitan Denise , Powell Bayard 
Published: April 2020

Abstract
Background: Acute myelogenous leukemia (AML) is a disease which largely affects older adults, for whom optimal therapy is unclear. Evidence based strategies are lacking to identify those older adults who may tolerate and benefit from standard therapies.
Objective: Test the feasibility of pre-treatment, inpatient geriatric assessment (GA) in older adults hospitalized with newly diagnosed AML.
Methods: Prospective evaluation of consecutive patients ≥60 years of age with newly diagnosed AML and planned induction chemotherapy admitted to a single institution from 6/2007–6/2008. Bedside GA was performed within 72 hours of diagnosis. In addition to demographics and routine labs, the GA measures obtained by a trained nurse included Mini-Mental Status Exam (MMSE), Center for Epidemiologic Studies Depression Scale (CES-D), Charlson Comorbidity Index (CCI), Vulnerable Elders Survey-13 (VES-13), Short Physical Performance Battery (SPPB, includes timed 4 meter walk, chair stands, standing balance), and grip strength.
Measures to assess feasibility included:
recruitment; time to complete the assessment, and proportion completing entire GA battery.
Results: Among 22 eligible inpatients, 11 enrolled (50%). The median age was 71 (range 63–78) and 72.7% were female. Poor risk cytogenetics were present in 27.3%. Laboratory measures included white blood cell count (mean=24.6×103/mm3, SD 26.2×103), hemoglobin (mean=8.5 g/dl, SD 1.6), platelet count (mean=60.3×103/mm3, SD=38.4×103), lactate dehydrogenase (LDH) (mean=340.0 U/L, SD=235.1), and albumin (mean=3.2 g/dl, SD=0.36). 66.6% of participants completed the entire GA battery; the remainder completed only self-report measures. Mean time for completion of the GA was 36.8 minutes (SD 9.8).
Mean scores for survey measures included: MMSE=26.1 (range 21–30, SD 3.2), CCI=1.6 (range 0–4, SD 1.1), CES-D=22.9 (range 8–37, SD 11.0), VES-13 survey=5.0 (range 1–8, SD 2.5). A wide range of objective physical performance was demonstrated including mean SPPB total score=7.4 (range 5–10; SD 2.1), mean timed 4 meter walk=12.9 seconds (range 6.6–22.1; SD 5.1), and mean grip strength=29.3 (range 12–62; SD 16.8).
Conclusions: Inpatient GA including physical performance assessment is feasible in older adults hospitalized for AML and our preliminary findings demonstrate significant variability in cognitive, emotional and physical status. These measures may represent valuable candidate predictors of outcomes, and ongoing studies will identify which measures are most predictive of treatment morbidity and response.
CLL-1, a Receptor Expressed on Myeloid Leukemic Stem Cells, Is a Potential Target for Immunotherapy of AML
Authors: Korver Wouter , Zhao Xiaoxian , Singh Shweta , Pardoux Cecile , Zhao Jingsong , Sen Siddhartha , Guzman Monica L. , Jordan Craig T. , Hsi Eric D , Abo Arie 
Published: April 2020

Abstract
CLL-1 (C-type Lectin-Like Molecule-1) is an inhibitory receptor expressed on myeloid cells which was previously shown to be expressed on AML cancer stem cells. To further validate the potential therapeutic against CLL-1, we generated a series of monoclonal antibodies (mAbs) against CLL-1 and assessed their cytotoxic and anti-cancer activities. While expression of CLL-1 was restricted to myeloid cells, it was expressed in 80% (37/46) of AML blasts but not in ALL blasts (n=5). Expression on AML CD34+/CD38- stem cells was detected in 71% (12/17) of cases. Selected anti-CLL-1 mAbs mediated dose-dependent. Complement Dependent Cytotoxicity (CDC) against various AML-derived cell lines with no detectable cytotoxic activity against lymphoid derived cell lines. Moreover, human embryonic kidney 293 cells became susceptible to anti-CLL-1 mAb mediated killing only after transfection with CLL-1, demonstrating that cytotoxic activity is mediated through a specific interaction with CLL-1. Furthermore, anti-CLL-1 mAbs showed CDC activity against all AML blasts tested in ex vivo assays (n=13), while no activity was observed against ALL blasts. CLL-1 efficiently internalizes upon antibody binding in both 293 transfected cells as well as AML cell lines, demonstrating the potential therapeutic opportunity for toxin-conjugation of anti-CLL-1 mAbs. In vivo anti-cancer activity of the lead chimeric mAb against CLL-1 was tested in an HL-60 xenograft model. In this model, growth of established tumors was reduced by 38% at 5 mg/kg twice weekly dosing. Our results demonstrate CLL-1 as an attractive target for AML with restricted expression on cells from myeloid origin, AML blasts and leukemic stem cells, as well as specific cytotoxic activity in in vitro, ex vivo and in vivo assays. We are currently undertaking additional xenograft models to evaluate the therapeutic potential of these mAbs against primary AML engrafted cells and in combination with chemotherapy in vivo.
Improvement of Remission Duration with Low-Dose Post-Remissional Gemtuzumab Ozogamicin (GO) in Elderly Patients with Acute Myeloid Leukemia
Authors: Marmont Filippo , Audisio Ernesta , D’Ardia Stefano , Salvi Flavia , Tonso Anna , Ferrero Dario , Levis Alessandro , Vitolo Umberto 
Published: April 2020

Abstract
The prognosis for elderly patients with acute myeloid leukaemia (AML) is dismal. Intensive chemotherapy is offered only to fit patients. The complete remission is obtained usually in 50% of the patients. The remission duration is short and the majority of patients will relapse within one year, with a poor quality of life. In this study we have evaluated the outcome of sixty-eight elderly patients (mean age 69.2 years) with AML or high-risk myelodysplasia (HR-MDS) treated with an induction program with fludarabine (30 mg/sqm/day for 4 days), cytarabine (1 gr/sqm/day for 4 days) and interspaced continuous infusion of idarubicin (12 mg/sqm/day on days 1,3,5), followed by lenograstim (FLAIG). Thirty-six patients (53%) were in CR after the first course. Thirty-four received a second reduced outpatient course. To account for the “guarantee time” required to survive the induction and retain the CR until the beginning of the post-remissional treatment, we used a landmark analysis at three months from CR to evaluate the disease-free survival (DFS), therefore excluding from the analysis early relapses and deaths in CR. In the first cohort, 13 patients received a post-remissional treatment with thioguanine and weekly cytarabine until relapse. In the second cohort, 14 patients received gemtuzumab ozogamicin (GO) at 3 mg/sqm every 3 months for 1 year or until relapse. The two cohorts were comparable for age, WHO classification, karyotype and CD33 expression. No difference in the median overall survival (OS) was observed between the two cohorts (7.4 vs. 12.3 months; p=0.1).. The landmark analysis showed an improvement of the DFS in patients receiving a post-remissional treatment with GO in confront to those treated with maintenance chemotherapy. (12.5 vs. 6.73 months; p=0.04). Although not influencing survival, the prolongation of DFS without hospitalisation or complications may improve the quality of life of elderly AML patients. The post-remissional therapy with low-dose GO deserves further evaluation in prospective randomized studies.
The Influence of Lepirudin, Bivalirudin and Dabigatran on the Calibrated Automated Thrombogram (CAT)
Authors: Woodhams Barry J , Carlo Audrey , Le Flem Lena , Guinet Celine , Depasse Francois , Samama Michel Meyer 
Published: April 2020

Abstract
In a two centre study (laboratories in Diagnostica Stago and Biomnis) we compared the in vitro effect on thrombin generation (TG) of Dabigatran and Bivalirudin (reversible direct anti-IIa inhibitors) with that of Lepirudin (an irreversible direct anti-IIa inhibitor) spiked into normal pool plasma. The effect of Lepirudin, Bivalirudin and Dabigatran were evaluated in both centres using the CAT (Diagnostica Stago, France) TG method in a concentration ranges up to 5, 20 and 1 μg/mL respectively. Testing was done in triplicate and repeated over 2 days. To reduce assay variability both centres used the same reagents lots and the same normal pool plasma (George King, USA). The range of each drug tested extended well above the therapeutic range concentrations normally found in patient plasma (0.5 to 1.0 μg/mL, 5 to 10 μg/mL and 0.1 to 0.3 μg/mL respectively for Lepirudin, Bivalirudin and Dabigatran). To see the effect of increasing activation forces, TG was performed at 3 different final concentrations of Tissue Factor (TF) - 1, 5 and 20 pM. All reagents were used as recommended by the manufacturer (Thrombinoscope, The Netherlands). A prolongation in the lag time (LT) is observed with all 3 drugs with all 3 concentrations of TF, but this is more marked for Lepirudin and Bivalirudin than it is for Dabigatran. In the therapeutic range Dabigatran (at 5pM TF) shows both an increase in LT and a decrease in peak thrombin and the ETP. At low concentration of Bivalirudin or Lepirudin, there is a paradoxical increase in peak height, which is even more pronounced at low TF concentration. At 1pM TF, this paradoxical peak increase is also observed with Dabigatran.
Results obtained in both laboratories are similar and complement our previous results and those reported elsewhere (1–4). The effect of Lepirudin and Bivalirudin on TG is different from that of Dabigatran. We also note that at lower TF concentration the anticoagulant effect on TG initiation is more intense but the test becomes less reproducible.
ANTI-hrs associated with the RHD-RHCE HAPLOTYPE DAR-ceAR in a patient with Sickle Cell Trait
Authors: Costa Sidneia Sanches , Chiba A. K , Martin F. O , Langhi Dante M , Chiattone C. S , Bordin Jose Orlando 
Published: April 2020

Abstract
Introduction: The RH blood group system is the most polymorphic and immunogenic among blood groups and some rare Rh phenotypes are found exclusively in the black population. The RH 18 phenotype (Hr-negative) that is one of these phenotypes is characterized by a high incidence antigen. The production of anti-RH 18 imposes to use equivalent rare antigen-negative red blood cells (RBCs), or deleted-Rh RBCs (Rhnull or D- -) for transfusion.
Case Report: A 24-year-old female patient with two previous abortions and one ectopic pregnancy and miscarriage had to be transfused with a RBC unit. She was typed as blood group A, Dccee phenotype with weak D, positive RBC antibody screening, auto-control negative and TAD negative. The antibody was anti-e (title 1/64) determined by testing serum against a commercial RBC panel (Fresenius-Kabi) by IgG indirect antiglobulin test tube (PEG-IAT), and nontreated (LissCoombs) and papain-treated RBCs on a gel matrix (DiaMed, Latino America). After adsorption of serum on Rhe-negative RBC (DccEE), the retrieving antibody was an anti-e-like antibody reacting with all normal Rhe-positive RBCs. Molecular and serologic Rh typing was also performed on blood samples obtained from five relatives of the patient and her husband.
Results: Patient and relatives cDNA sequence of gene RHD and RHCE transcripts were analyzed for RHCE exons 4 to 5 and exon 6; for RHD exons 4 to 5, 6 and 7. Sequencing of cDNA from the patient, her father and her sister showed in RHD the presence of the DAR allele point mutation 602 C&gt;G in exon 4; 667 T&gt;G and 744 C&gt;T in exon 5; 957 G&gt;A and 1025 T&gt;C in exon 7; while on gene RHCE we found the ceAR allele carrying 712 A&gt;G, 733 C&gt;G e 787 A&gt;G in exon 5. Hemoglobin tests revealed that the patient, her father and sister had a sickle cell trait (patient HbA1=55%; HbA2=2,8%; HbS=41%; HbF=1,2%; father HbA1=58,5%; HbA2=1,4%; HbS=37,7%; HbF=2,4%; sister HbA1=57,4%; HbA2=2,5%; HbS=38,6%; HbF=1,5%).
Discussion: The family study showed that the haplotype DAR-ceAR was inherited from her father. Individuals carrying the ceAR allele may produce anti-hrs antibody that is defined as a Rhe-like which does not react with rare Rhe positive variant RBCs. The antibody may be clinically significant and represents a great risk for those patients who are not transfused with a similar rare phenotype variant. Serologically, DAR has shown weaker reactions with anti-D, it is characterized by complete loss of at least 9 of 37 RhD epitopes, and may produce anti-D. Both mutated alleles show a replacement of RHD exons by RHCE counterparts.
An Investigation Report of Frozen Plasma Evidence-Based Transfusion in a Tertiary Care Teaching Hospital
Authors: Chen Qin-fen , Li Pei , Xie Yi , Chen Bao-An 
Published: April 2020

Abstract
Objective: To estimate the appropriateness of frozen plasma (FP) transfusions in a tertiary-care teaching hospital.
Methods: The medical records of all hospitalized patients transfused with FP between 1 April 2007 and 31 April 2008 were reviewed, analyzed retrospectively according to the Clinical Blood Transfusion Technical Specification from Ministry of Healthy.
Results: In 5084 times 13627 units FP transfusions, departments of surgery used 77.7%, and departments of internal medicine used 22.3%. The medical records showed that the reasons of FP transfusion included supply coagulation factors, supply intraoperative blood loss, postoperative supporting therapy for enhancement of wound healing, volume expansion, plasma exchange, nutritional support and massive transfusion. Overall, the 68.5% of FP transfusions were assessed as inappropriate. The inappropriateness rate in departments of surgery was 71.7%, and that in departments of internal medicine was 22.6%.
Conclusion: There is considerable inappropriate transfusion of FP in our hospital, for there has some important objective reasons. It is stated that the Technical Specification is not suitable to the condition of blood production supply and clinical practice. Clinicians should follow evidence-based transfusion guidelines for appropriate use of FP. So we suggest the Technical Specification should be renew periodly.
Assessment of SBDS Expression Patterns in Human Marrow Hematopoietic and Stromal Cells by Immunohistochemistry and Its Use as a Diagnostic Screen for Schwachman-Diamond Syndrome
Authors: Wong Trisha , Calicchio Monica , Fleming Mark D. , Shimamura Akiko , Harris Marian Hester 
Published: April 2020

Abstract
Objective. Shwachman Diamond Syndrome (SDS) is an autosomal recessive syndrome characterized clinically by exocrine pancreatic insufficiency, bone marrow dysfunction, skeletal abnormalities and a predisposition to leukemia. Molecularly, it is characterized by mutations in the Shwachman-Bodian-Diamond-Syndrome (SBDS) gene encoded on chromosome 7. The majority of mutant alleles (74%) associated with SDS are the result of gene conversion events with an adjacent SBDS pseudogene, leading to truncation, splice site, and frameshift mutations and a marked decrease in SBDS expression. The remaining pathogenic alleles include frameshift and, rarely, missense changes1. In this study, we examined the SBDS expression pattern in bone marrows from patients with SDS compared to normal controls or patients with other bone marrow failure syndromes using a new immunohistochemical (IHC) assay.
Methods. Archived bone marrow biopsy tissue sections from patients with SDS (21 specimens from 9 patients), idiopathic aplastic anemia (AA, 12 patients), Fanconi Anemia (FA, 4 patients), Diamond-Blackfan Anemia (DBA, 9 patients), Severe Congenital Neutropenia (SCN, 3 patients) and 13 normal controls were stained with an antibody directed against the C-terminus of SBDS. Four cell lineages (myeloid precursors, megakaryocytes, plasma cells and osteoblasts) were scored in a blinded fashion by two pathologists on a scale from 0 (negative) to 3 (strong). The scores from each cell lineage within each diagnostic group were averaged to yield an overall IHC score.
Results. We first characterized the SDS expression pattern in normal controls. Promyelocytes, early myelocytes, megakaryocytes, plasma cells, and osteoblasts all showed high levels of SBDS protein. In the myeloid lineage, SBDS expression decreased to undetectable levels in mature granulocytes, consistent with prior reports that SBDS is not necessary for terminal neutrophil differentiation. SBDS staining was not detectable in erythroid precursors or lymphocytes. We next compared SBDS IHC staining in these normal controls to that seen in SDS patients and patients with other bone marrow failure syndromes. The mean overall IHC score for SDS patients was 0.6 whereas it was 2.5 in normal controls, 2.23 in AA, 2.4 in DBA, 2.67 in FA and 2.38 in SCN. Four samples from two SDS patients with rare missense mutations in SBDS had moderate staining, consistent with previous studies that showed intermediate levels of SBDS expression in patients with missense mutations. When the analysis was restricted to patients harboring biallelic gene conversion SBDS mutations, the mean SBDS IHC score dropped to 0.14.
Conclusion. SBDS protein is highly expressed in early myeloid progenitors, megakaryocytes, and osteoblasts. This novel SBDS IHC assay may provide a rapid screen for the most common SBDS mutations.