bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect Of Purified Poloxamer 188 and Various Dextrans On Erythrocyte Sedimentation Rate In Healthy Subjects and Patients With Sickle Cell Disease
Authors: Hoppensteadt Debra , Emanuele Martin , Molnar Joann , Iqbal Omer , Fareed Jawed 
Published: October 2019

Introduction
Purified poloxamer188 (P188) (Mast Therapeutics) is a non-ionic, linear block copolymer composed of a central chain of hydrophobic polyoxypropylene and two flanking chains of hydrophylic polyoxyethylene (MW 8.5 kDa). This agent has hemorheologic properties which result in improved microvascular blood flow. P188 has been investigated in a number of indications and is currently under study in an international phase 3 clinical trial in sickle cell patients with vaso-occlusive crisis. Dextrans represent branched polysaccharides of 10-70 kDa that have been used as antithrombotic agents and plasma expanders. Sickle cell disease (SCD) represents a complex hemorheologic condition due to RBC aggregation and cell-fibrin/fibrinogen interactions. The erythrocyte sedimentation rate (ESR) is reflective of RBC and plasma interactions. This study was designed to compare the effect of P188 and dextrans on ESR’s in blood obtained from healthy subjects and patients with sickle cell disease who were seen at Loyola University Medical Center clinics.

Material and Methods
Whole EDTA blood collected from normal individuals (n=8) and sickle cell patients confirmed by electrophoresis (n=11) were supplemented with P188 or dextran 10K, 18K , 40K and 70K at various concentrations (or saline control). ESR was measured using standard laboratory technique.

Results
The ESR’s for sickle cell patients (26.4 ± 7.1 mm/hr) were significantly higher in comparison to the ESR’s for healthy subjects (14.6 ± 2.1 mm/hr). Supplementation of P188 decreased ESR’s in both populations. Normal blood ESR’s decreased to 9.1 ± 1.3 mm/hr (38%), whereas the sickle cell patient values decreased to 14.1 ± 4.6 mm/hr (47%). At comparable concentrations, none of the dextrans changed ESR’s in healthy subjects or patients with sickle cell disease.

Discussion
These results demonstrate that ESR in SCD patients are elevated compared to healthy subjects. P188 supplementation decreased (up to 50%) ESR’s in both the healthy subjects and sickle cell patients. This may be due to the inhibition of rouleaux formation resulting from P188 effects on RBC membranes or cell-protein interactions. None of the dextrans produced a similar decrease, suggesting that the observed lowering of ESR by P188 is unlikely to be due to a non-specific effect related to polymer molecular weight.

Clinical Implications
P188 is a potential therapeutic agent which may facilitate blood flow and reduce cell-fibrin/fibrinogen interactions in a variety of hemorrheologic disorders. The observed decrease in ESR both in normal and sickle cell blood samples by P188 may primarily be due to increased membrane hydration, fibrinogen dispersion and anti-adhesive effects of this agent.

Disclosures:
Emanuele: Mast Therapeutics: Employment. Fareed:Mast Therapeutics: Research Funding.
Decreased Levels Of Procoagulant Phospholipids In Bleeding Patients With Overcoagulation By Vitamin K Antagonists
Authors: Van Dreden Patrick , François Dominique , Mathieu Emmanuel , Grusse Matthieu , Vasse Marc 
Published: October 2019

Background
The major adverse effect of vitamin K antagonists (VKA) is the increased risk for bleeding complications. In addition to well-identified risk factors such as age, prior gastrointestinal tract bleeding, or hypertension, the intensity of anticoagulation evaluated by the International Normalized Ratio (INR) measurement is a major determinant of VKA-induced bleeding. However, for the same degree of VKA overcoagulation and comorbidities, some patient will bleed, whereas others remain asymptomatic. Therefore, identifying specific biological markers that identify patients at high risk of bleeding would have great clinical impact. Microparticles, derived from different cellular origins (endothelium, red blood cells, leukocytes, platelets or apoptotic tissues), can be detected in plasma and express procoagulant phospholipids (PPL). The presence of PPL has been associated with various diseases complicated by an hypercoagulable state. Therefore, we hypothesize that the procoagulant activity of PPL could protect against haemorrhage in patients with VKA overcoagulation.

Patients and methods
53 consecutive patients who were referred to the emergency department of our institution and with an INR > 5 were enrolled in the study: 22 (10 females; 12 males, median age 82 years) were symptomatic (20 cases of minor bleeding, 2 cases of non-fatal major bleeding), whereas 31 (18 females, 13 males, median age 78 years) were asymptomatic. Median INR was 7.36 (range: 5 – 22.6) and 6.3 (range: 5 – 10.7) in symptomatic and asymptomatic patients, respectively (p = 0.17, not significant). PPL were evaluated using a factor Xa-based coagulation assay (STA-Procoag-PPL, Diagnostica Stago) in which shortened clotting times are associated with increased levels of PPL. We also quantified thrombomodulin (TM) by an ELISA assay (Asserachrom Thrombomodulin, Diagnostica Stago) and by a functional assay based on the ability of TM to activate Protein C in the presence of thrombin, since high plasma levels of TM were previously identified as a predictor of bleeding complications.

Results
Clotting times were significantly lower in asymptomatic patients than in bleeding patients [respective median values 36.5 seconds (range: 27.1 – 72.2) and 47.2 seconds (range : 30.5 – 72.8); p = 0.03. In contrast, there were no significant differences for TM levels, whatever the assay used (functional or immunological).

Conclusion
Increased PPL could contribute to decrease the haemorrhagic risk of patients treated by VKA. It is not clear if the decrease of PPL is directly responsible of the hemorrhagic syndrom, or if PPL are decreased because of a consumption during the hemorrhagic episode. In order to answer this question, it could be of interest to analyse if a prospective follow-up of this parameter could help to identify patients with an increased hemorrhagic risk when treated by VKA.

Disclosures:
No relevant conflicts of interest to declare.
Effect Of Recombinant Human Soluble Thrombomodulin On Clinical Outcomes Of patients With Coagulopathy After Hematopoietic Stem Cell Transplantation
Authors: Chi SungGi , Ikezoe Takayuki , Takeuchi Asako , Anabuki Kazuki , Kim Tsukie , Sakai Mizu , Taniguchi Ayuko , Togitani Kazuto , Yokoyama Akihito 
Published: October 2019

Recombinant human soluble thrombomodulin (rTM) comprises the active, extracellular domain of thrombomodulin (TM), and inactivates coagulation by binding to thrombin. In addition, the thrombin-rTM complex activates protein C to produce activated protein C (APC), which inactivates factors VIIIa and Va in the presence of protein S, further inhibiting thrombin formation. The use of rTM for the treatment of DIC was approved in Japan in in 2008, and since then it has proved effective in individuals with DIC complicated by a variety of underling diseases including sepsis and acute leukemia. Of note, rTM possesses anti-inflammatory and cytoprotective effects and the use of rTM successfully rescued individuals with sinusoidal obstruction syndrome, engraftment syndrome and transplantation-associated microangiopathy complicated by hematopoietic stem cell transplantation (HSCT). The present study retrospectively investigated whether the use of rTM improved the clinical outcome of individuals who received HSCT in our institution.
From 2001 to 2012, 71 individuals with hematological diseases received HSCT in our institution. Of these, 41 developed coagulopathy in association with various underlying conditions. The patients who developed coagulopathy after 2008 (n=23) were treated by rTM, and the others (n=11) were treated by either heparin and/or antithrombin III concentrate. Seven patients did not receive any anticoagulant therapy. Of note, treatment of coagulopathy by rTM significantly improved clinical outcomes of patients at day 100 and dramatically prolonged their overall survival (p=0.044). Taken together, rTM is useful to improve clinical outcomes of transplant recipients with coagulopathy.

Disclosures:
No relevant conflicts of interest to declare.
Platelet RNA-Seq Identifies Genes Expressed Differentially After Clopidogrel Treatment
Authors: Prakasa Kalpana Rao , Becker Lewis , Faraday Nauder , Becker Diane , Qayyum Rehan 
Published: October 2019

Background
Clopidogrel is an antiplatelet agent that acts through direct inhibition of ADP-dependent platelet activation. Several studies have also found an inhibitory effect of clopidogrel on inflammation. Although platelets are known to participate in inflammatory processes, the mechanism(s) through which clopidogrel might modify platelet-mediated inflammation is unknown. We examined the platelet transcriptome to gain insight into the mechanism(s) through which clopidogrel modulates inflammation.

Methods
Three individuals with subclinical atherosclerosis and family history of early onset coronary artery disease were enrolled. Blood was collected for RNA-seq at baseline and one-week after clopidogrel 75mg daily. RNA was extracted from leukocyte-depleted platelet-rich plasma and approximately 70 million 100bp paired-end reads per sample were obtained using the Illumina HiSeq 2500. RNA-seq analysis was performed using Bowtie/TopHat/Cufflinks software pipeline. Genes with expression ≥ 0.3 fragments per kilobase of transcript per million reads (FPKM) were considered to be expressed. We set the threshold for significant differential expression as ≥ 2-fold change in gene expression after clopidogrel therapy with a false discovery rate of 0.05.

Results
The study sample consisted of 2 women and 1 man. Among the 12,297 expressed genes, the most highly expressed nonubiquitous genes included PPBP, PF4, and TUBB1, confirming platelets as the source of the RNA. Comparing samples before and after clopidogrel, treatment was associated with up-regulation of 733 genes and down-regulation of 387 genes. Pathway analyses found that genes related to platelet activation, degranulation, and aggregation were up-regulated, while genes related to immune system processes, such as B- and T-cell activation and innate immune response, were down-regulated. Examples of down-regulated genes include the toll-like receptor family genes, TLR2, TLR4, TLR7, and TLR8, the chemokine receptor genes CCR2, CCR5,CCR6, and CCR7, and the interleukin receptor genes, IL4R, IL7R, and IL13RA.

Conclusion
One week of clopidogrel therapy up-regulates transcription of pro-thrombotic platelet genes and down regulates transcription of inflammation-related platelet genes. These data suggest that clopidogrel may exert a direct anti-inflammatory action by inhibiting the ability of platelets to participate in inflammatory processes. The relationship of variability in clopidogrel-induced modulation of these pathways to clinical outcomes is unknown.

Disclosures:
No relevant conflicts of interest to declare.
UV-C Treatment Of Intermediates Of Biologicals To Inactivate Viruses Without Denaturing Desired Protein
Authors: Groener Albrecht , Dreier Christine 
Published: October 2019

Dedicated virus reduction steps implemented in the manufacturing process of biologicals, either isolated from human plasma or produced as recombinant proteins, are essential safety measures to assure that a potential virus contamination of the source material will not be transmitted to patients requiring these therapeutic proteins.
Currently applied virus reduction steps as solvent/detergent treatment and virus filtration are very effective virus reduction methods with inherent method-dependent gaps regarding the reduction capacity for a very wide range of viruses of diverse physico-chemical characteristics: solvent/detergent treatment does not inactivate non-enveloped viruses and, depending on the pore size of the virus filter, small viruses are not removed when the desired protein is large and has to pass the filter. Therefore, another virus inactivation method was studied which is considered especially effective for small viruses: UV-C treatment using the UVivatec system provided by Sartorius Stedim Biotech GmbH, Göttingen, Germany.
Experiments were performed to study the impact of UV-C treatment on the integrity of proteins employing fibrinogen as an example for a large protein and on the inactivation capacity for poliovirus (a small non-enveloped virus). The integrity of fibrinogen was assessed by comparing the untreated fibrinogen with the UV-C treated fibrinogen using HPLC, Clauss assay and thromboelastometry. Virus inactivation was studied in a bioassay using a sensitive cell culture infectivity assay employing a cynomolgus cell line.
The results show that UV-C treatment inactivates viruses and modifies fibrinogen in a dose dependent manner; the monomer, dimer and polymer peak in the fibrinogen preparation studied changed from approx. 75% to 60%, 17% to 25% and 8% to 13%, respectively, at a UV-C intensity of 400 J/m² demonstrated by HPLC measurement. In order to protect fibrinogen from modifications, the antioxidant glutathione was added to the fibrinogen preparation. At an UV-C intensity of approx. 300 J/m², sufficient to effectively inactivate viruses studied, a modification of fibrinogen was not any longer detectable.

Disclosures:
Groener: CSL Behring: Employment.
Are We Appropriately Using Intravenous Immunoglobulin In The Saskatoon Health Region?
Authors: Manna Mita , Karim Safiya , Brose Kelsey , Dallas Karen L 
Published: October 2019

Intravenous Immunoglobulin (IVIG) is a plasma protein used in the treatment of various diseases. The cost of IVIG is approximately $50-$80 per gram. In 2005, Saskatchewan's usage of IVIG was above the national average, at 100g per 1000 persons compared to 95g per 1000 persons. IVIG is currently licensed by Health Canada for use in six diseases; however, IVIG is often used for off-label indications. There are several published guidelines that outline recommended uses of IVIG. These are largely based on case studies and expert opinion. Despite these guidelines, IVIG is often used in multiple settings where there is no supporting evidence.
We aimed to evaluate adherence to published guidelines by quantifying IVIG usage in the Saskatoon Health Region (SHR). We conducted a retrospective chart review of patients who received IVIG in 2011 and 2012, classifying the usage based on diagnosis and prescribing specialty. Our primary endpoint was the percentage of appropriate use of IVIG in comparison to published guidelines. Our secondary endpoint was adverse reactions seen in patients receiving IVIG.
In 2011, 227 patients received IVIG totalling 50,528g and 2.78 million dollars. In 2012, 216 patients received IVIG totalling 63,155g and 3.49 million dollars. According to published guidelines, 36% of the total usage in 2011 and 21% of the total usage in 2012 was not indicated (Figure 1). IVIG was prescribed the most by neurologists, hematologists, and rheumatologists (Figure 2). Despite the recent guidelines, the results show that there was no significant improvement in usage of IVIG between 2011 and 2012 (p-value 0.079, odds ratio 1.46). No adverse reactions were documented.
In conclusion, our data analysis shows that IVIG usage in the SHR does not adhere well to guidelines. Pre-printed order sets outlining appropriate indications for IVIG are currently being introduced in the SHR to help improve adherence to guidelines. After the order sets have been implemented for one year, we will re-evaluate the usage of IVIG in the SHR to see if improvements have been made. Our goal is to ensure there is appropriate resource allocation of IVIG and to reduce risks to the patient and financial costs to the health care system.

Disclosures:
No relevant conflicts of interest to declare.
A New Method For Electronic Crossmatch: ABO /Rh Blood Group Confirmation and Antibody Screening Concomitantly With Serologic Crossmatch
Authors: Demirkan Fatih , Gunal Veli , Dereli Yasar 
Published: October 2019

Introduction
Altough electronic crossmatch (E-XM) is cost effective and reduce laboratory workload, practical difficulties of determining 2 consecutive ABO/Rh in patients, requirement for the verification of the donor ABO group, concerns for errors in patient identification/specimen labeling and lack of reimbursement for performing ABO confirmation testing on a second separately collected specimen could avoid its widespread implementation in some countries.

Method
Using gel santrifuge method, ABO/Rh grouping and immuno-hematologic tests are done by a fully automatic system and recorded in blood bank information system. For patients and donors who do not have a previous blood group and antibody screen in the system, study steps are: 1. ABO/Rh group is performed with the 1st specimen and recorded 2. Donor blood group and antibody screen are done in the same gel card. 3. A second specimen is required at the time of transfusion request for regular antiglobulin crossmatch and antibody screen. ABO confirmation of patient and donor is done concomitantly in the same card with XM. 4. When 2 or more units are requested, first unit is selected by serologic XM, subsequent units are selected by E-XM from the available donor pool. Donor pool for electronic selection is fed by serologically compatible but undelivered units. Computer system is capable of questioning availability for E-XM eventhough serologic XM is not performed and could show the missing parameters needed for E-XM.

Results
After 6 months of validation process, between 1.01.2012- 31.07.2013, 36.7% of erythrocytes were reserved by E-XM using this method (table 1). Evaluation of the efficacy of the XM process revealed that 37.5% of the serologically confirmed units were undelivered and those supported the pool for E-XM (table 2).

Conclusions
1. This method implements a double specimen system for ABO/Rh verification to ensure the transfusion security and guarantees the selection of at least 1 unit serologically. 2. Reduces work load and cost effective even in centers who are currently using serologic XM.

Disclosures:
No relevant conflicts of interest to declare.
Inhibition of hydroxymethylglutaryl coenzyme A reductase activity induces a paradoxical increase in DNA synthesis in myeloid leukemia cells
Authors: Hohl RJ , Larson RA , Mannickarottu V , Yachnin S 
Published: October 2019

Abstract
The effects of competitive inhibition of hydroxymethylglutaryl coenzyme A (HMG CoA) reductase by compactin on the in vitro proliferation of peripheral blood myeloid leukemia cells were studied using the cells from 45 patients with acute myeloid leukemia or chronic myelogenous leukemia in blast phase. The cells from 58% of these patients showed a dose-related inhibition of DNA synthesis when incubated with compactin. Unexpectedly, cells from 18% of the patients were resistant to the inhibitory effects of compactin on DNA synthesis and responded to the HMG CoA reductase inhibition with an actual increase in the incorporation of 14C-labeled thymidine into DNA. Another 18% of the patients studied displayed both inhibition and stimulation of DNA synthesis in a biphasic response depending on the particular concentration of compactin used. The maximum enhanced rates of cellular DNA synthesis were observed with lower compactin concentrations (5 x 10(-7) mol/L) than were required for maximum inhibition of DNA synthesis (10(-5) mol/L). Leukemia cells displaying a stimulated response to compactin had a significantly lower baseline DNA synthetic rate than did cells that showed an inhibitory response of DNA synthesis to compactin. There was no correlation between these cells' varying DNA synthetic response to compactin and measures of baseline HMG CoA reductase activity or acetate conversion to cholesterol. Whereas the observation of cellular DNA synthesis stimulation by HMG CoA reductase inhibition has not been observed in other mammalian cells and seems paradoxical, explanations may emerge in light of our growing knowledge concerning the importance of isoprenylation for the function of certain cell regulatory proteins.
Differentiation in B-precursor acute lymphoblastic leukemia cell populations with CD34-positive subpopulations
Authors: Re GG , Estrov Z , Antoun GR , Felix EA , Pinkel DP , Zipf TF 
Published: October 2019

Abstract
B-precursor acute lymphoblastic leukemia bone marrow specimens that contained subpopulations of cells with immunophenotypes corresponding to early (CD34) and late (CD20) and (CD22) stages of normal B-cell differentiation were studied. Subpopulations of cells were isolated according to immunophenotype and then analyzed by both a clonogenic assay and molecular genetic methods. Clonal equivalence of the early and late immunophenotypic subpopulations was confirmed for each case by the demonstration of identical lg gene rearrangements. The in vitro colony-forming assay consistently showed a growth advantage for the CD34+ subpopulations over the CD34- subpopulations. CD34 mRNA was detected readily in these isolated precursor cells. When two specimens in which virtually all of the leukemia cells were CD34+ and CD34+CD20+ and CD34+CD22+ subpopulations were also present the CD34 mRNA was limited to the cells without the late-stage differentiation antigens on their surface. Furthermore, the c-myb mRNA was found only in the subpopulations that also contained CD34 mRNA. Our results show that a limited program of differentiation reminiscent of normal B-cell development may be present in this leukemia.
Induction of HL60 cell differentiation by tiazofurin and its analogues: characterization and efficacy
Authors: Goldstein BM , Leary JF , Farley BA , Marquez VE , Levy PC , Rowley PT 
Published: October 2019

Abstract
Among inducers of myeloid differentiation for leukemic cells, tiazofurin is of special interest because its mechanism of action is known; it inhibits inosine monophosphate dehydrogenase and thus decreases the guanine nucleotide pool. Reported here are three aspects of tiazofurin induction of myeloid differentiation in HL60 human acute promyelocytic leukemia cells. First, inductive efficacy was evaluated for analogues ara-tiazofurin, xylo-tiazofurin, and selenazofurin, for dinucleotide anabolites thiazole-4-carboxamide adenine dinucleotide (TAD) and selenazole-4-carboxamide adenine dinucleotide (SAD), and for a phosphodiesterase-resistant TAD analogue, beta-methylene TAD. The results showed that the parent compounds are more effective inducers than the dinucleotide derivatives and that the selenazole analogues are more effective inducers than the thiazole compounds. Second, HL60 cell induction by tiazofurin was shown to be synergistic with that produced by the antiviral agent ribavirin. Finally, tiazofurin was found to induce expression of a phosphatidylinositol-specific phospholipase C- sensitive Fc gamma-receptor III (FcRIII) on HL60 cells, a feature consistent with neutrophilic, but not monocytic, differentiation.