bluuebunny/crossref_metadata_2025_split huggingface dataset:
Factor V G1691A Mutation and Factor II G20210A Variant in Children with Venous Thrombosis or Stroke. An International Database Update.
Authors: Nowak-Gottl Ulrike , Kurnik Karin , Manco-Johnson Marilyn , Young Guy 
Published: April 2020

Abstract
The present database pooling was performed to investigate the clinical phenotype in pediatric factor V G1691A (FV) and factor II G20210A (FII) carriers. 230 pediatric patients with deep venous thrombosis (DVT: n=143) or arterial ischaemic stroke (AIS: n=94) carrying either the FV (n=178) or the FII variant (n=52) not suffering from additional thrombophilic risk factors were consecutively enrolled from 2 US centers and one German database. Antithrombotic therapy was performed according to standard protocols either with LMWH/vitamin K-antagonists (DVT) or LMWH/ASS (AIS) for 6 months (range: 3–indefinitive). Patients were followed for a median of 5.4 years (range: 1–23). FV children with first disease onset were significantly younger compared with patients in the FII group (p=0.02), and showed more abdominal vein thrombosis than children with the FII variant (p=0.02).
Following withdrawal of antithrombotic therapy a first recurrent event was diagnosed in 12 of 178 children with the FV mutation (6.7%), whereas pediatric patients with the FII mutation showed recurrent thrombosis in 6 of 52 patients (11.5%). Median (range) relapse time was 6.5 years (0.1–15.2) in FV carriers compared with 15 years (13–16) in the FII group (p=0.06). Interestingly, recurrent thrombotic events were recorded significantly more often in children with DVT compared with AIS (p&lt; 0.001).
In conclusion, the clinical phenotypes of FVC and FII are different with respect to first thrombotic onset, and thrombotic locations, and recurrent thrombosis in both phenotypes occurred more often in DVT compared with AIS.
IgA Anticardiolipin Antibodies as a Risk Factor for Arterial and Venous Thrombotic Events.
Authors: Duran Esther Urbaez , Kasunic Tim , Cotant Matthew , Kuriakose Philip 
Published: April 2020

Abstract
Background: Many studies have concluded that the presence of anticardiolipin antibodies is associated with both venous and arterial thrombotic events. The role of IgA anticardiolipin has been studied previously with conflicting results. Some of this disparity is thought to be due in part to the lack of international standards in measuring these levels.
Objective : To further evaluate the role of IgA anticardiolipin anitbody as a risk factor for thrombosis.
Patients and Methods: We conducted a single institution retrospective review of all patients with elevated IgA anticardiolipin antibodies extracted from a database of 20,000 patients for whom IgA anticardiolipin had been measured.
Results: One hundred forty-eight patients were identified with elevated IgA anticardiolipin antibodies as measured by semiquantitative indirect Elisa. Patients had a median age of 52.5; 52% were African American, 38% Caucasain and 9% of other ethnic background. Of these patients, 67 had associated arterial or venous thrombotic events: 15.9% were pulmonary embolisms, 47.5% were deep venous thromboses and 36.6% were arterial thrombotic events. Patients were further evaluated for other thrombotic risk factors including IgG and IgM anticardiolipin antibodies; lupus anticoagulant; pt 20210; activated protein C resistance/Factor V Leiden; hyperhomocysteinemia; antithrombin; protein C/protein S deficiency; active malignancy with or without concurrent chemotherapy; tamoxifen, oral contraceptive or hormone replacement therapy; catheter-related risk; pregnancy; myeloproliferative disorders and hyperviscosity syndromes; sickle cell disease; and surgery, trauma or immobilization within 6 weeks of the event. In 35.8% of evaluated patients with elevated IgA and thrombotic events, there was was no concommitant elevation of IgG or IgM anticardiolipin. In 33.3% of patients with thrombotic events, none of the other predetermined risk factors were present.
Conclusion: IgA anticardiolipin may be an independent risk factor for development of venous and arterial thrombotic events. A large case-controlled or prospective, randomized trial with standardized measurement will likely be needed to further clarify this issue.
Sticky Platelet Syndrome: The Clinical Spectrum of Thrombosis.
Authors: Bick Rodger L. , Fareed Jawed 
Published: April 2020

Abstract
Sticky Platelet Syndrome (SPS) was first described by Mammen, E., et al. In 1984, is inherited as an autosomal dominant and is associated with both arterial and venous thrombosis. It is also associated with placental vascular thrombosis and resultant recurrent miscarriage syndrome (RMS). It was thought to be of interest to assess the prevalence and types of thrombosis associated with SPS in a strictly thrombosis hemostasis referral practice over a one year period. During 2004, 599 first-time new thrombosis patients were seen, excluding recurrent miscarriage syndrome (RMS). Diagnosis of SPS was by the method of Mammen. Of 599 pts. 20.5% had SPS. The most common defect in the population was antiphospholipid syndrome and Factor V Leiden was seen in 8.5% of all pts. Of these SPS patients 60% suffered arterial (AA) and 39% suffered venous thrombosis. 4 pts. Had both. The types of thrombosis in descending order of prevalence were as follows: DVT = 27.6%, cerebrovascular thrombosis (CVT) = 19.5%, transient ischemic attacks (TIA) = 18.6%, PE = 12.1%, retinal artery thrombosis (RT) = 8.1%, mesenteric AA thrombosis = 7.3%, microvascular peripheral AA = 7.3%, coronary AA = 3.2% and popliteal AA = 0.8%. All pts. Were treated with low-dose ASA and those with CVT or TIA also received clopidogrel (75 mg. /day). At one-year follow-up, no recurrences were seen.
In summary: as has been noted by Mammen and others, SPS is a common hereditary thrombophilia associated with arterial and venous thrombosis. The prevalence in this population of first-time thrombosis pts. Was 2-fold that of Factor V Leiden mutation. Thus, a work-up for SPS should always be considered in pts. With unexplained thrombosis.
Warfarin Resistance Due to a Val29Leu Mutation in the VKORC1 Protein.
Authors: Huff Jason D. , Lawson Heather L. , Harish V. C. , Zhang Lin , Owen John 
Published: April 2020

Abstract
Warfarin resistance is a common clinical problem encountered by physicians of all subspecialities. Medical noncompliance and dietary indiscretions are felt to be the usual causes of a subject’s “resistance” to anticoagulation with coumarin derivatives. Warfarin’s target is the vitamin K epoxide reductase complex (VKOR). This complex functions within the vitamin K dependent γ-carboxylase system to recycle oxidized vitamin K. Until recently, the genetic sequence of components of this complex remained a mystery (Li et al., Nature 2004). Mutations in a subunit of this complex named vitamin K epoxide reductase complex subunit 1 (VKORC1) were isolated in human subjects with warfarin resistance (Rost et al., Nature 2004). Knowledge of this coding sequence has enabled us to study individual subjects suspected to have a genetic source for their warfarin resistance. Our study subject is a 24-year-old Caucasian female who received warfarin after developing bilateral pulmonary emboli. She required 30 mg of warfarin daily to achieve an international normalized ratio greater than 2.0. After institutional review board approval and with the patient’s informed consent, genomic DNA was extracted from the subject’s peripheral blood. The three exons comprising the VKORC1 gene were amplified by polymerase chain reaction (PCR). PCR primers were designed using the published sequence and online Primer3 software. Prolonged extension times were required to obtain good amplification of all three exons. After purification, the PCR products were sequenced and analyzed. The sequence analysis demonstrated a heterozygous G→T mutation in exon 1 at base pair 85. This results in a Val29Leu substitution, which has been described in a warfarin resistant patient by Rost et al. The second and third exons matched completely with the published sequence. The gene frequency of the mutation was then determined by examining 400 random, discarded DNA samples. The HypCH4 IV restriction enzyme cleaves at 2 positions in exon 1 in the wild-type genotype, but at just one position if our known mutation is present. The mutation was not identified in any sample.
In conclusion, we have identified a second patient with a Val29Leu substitution in vitamin K epoxide reductase resulting in warfarin resistance. Our analysis of 400 random samples has not identified any other example of this mutation, indicating that this is a rare occurrence. Genetic analysis of patients requiring more than 20 mg of warfarin daily would likely reveal more mutations in the VKORC1 protein. The identification of such mutations would provide valuable insight into the structure-function relationships of this protein complex.
Rituximab Immunotherapy for the Antiphospholipid Syndrome.
Authors: Bern Murray M. 
Published: April 2020

Abstract
The antiphospholipid syndrome (APS) is a heterogeneous disease expressed in a variety of ways, including thromboembolic disease. Therapy for this phase of APS is primarily anticoagulation. When the disease is aggressive and resistant to anticoagulants immunotherpay has been applied with variable success. Steroids, cyclophosphamide, plasma exchange, plasmapheresis, and immunoglobulin infusion have been used. This is a report of a patient with very aggressive APS treated successfully with rituximab immunotherapy. This 60 year old female had four episodes of deep vein thrombosis and two episodes of pulmonary embolus. An inferior vena cava filter had been placed. She came to hematologic attention in 2001, when the APS diagnosis was established based upon positive lupus anticoagulant and antibodies to cardiolipin and phosphatidyl serine. Long term steroid therapy was considered potentially too toxic. Azathioprine therapy was initiated at 100 mg daily and then increased to 150 mg daily without reduction of antibody titers. Thereafter she developed thrombosis in-situ in the pulmonary artery, requiring surgical thrombectomy. Therapy with rituximab was begun at 375 mg/m2 weekly times four, with repeat infusions every 6 months. Infusions were well tolerated. Warfarin therapy was continued throughout. Antibody titers have fallen.
Antibody titers after 3 cycles of therapy Anticardiolipin Baseline (mg/dl) 3 cycles of Therapy Followup (mg/dl) IgG 1032 854 IgM 255 &lt;20 IgA 645 495 Antiphosphatidyl serine IgG 1620 775 IgM &lt;25 &lt;25 IgA 720 315
There have been no further episodes of TED. Therapy with rituximab has been well tolarted and appears to be effective. Therapy is continuing with additional antibody titers to be obtained.
Predictors of Red Cell Transfusion in Medically III Patients.
Authors: Yuen Maggie M. , Anderson David R. , Rose Caren 
Published: April 2020

Abstract
Although red cell transfusion is a relatively common treatment in medically ill patients, much is not known about the clinical determinants used to guide its use. We conducted a retrospective, case-controlled chart review of admissions to the medical teaching units (MTU) at this center (Queen Elizabeth II Health Sciences Centre, Halifax, Nova Scotia). Forty-two patients who received red blood cell transfusion during admission to the MTU from January to March 31, 2004. Clinical data collected included: age, gender, indication for MTU admission, co-morbid disease, smoking, admission hemoglobin, pre- and post-transfusion hemoglobin, arterial pressure of oxygen, lactic acidosis, parameters of shock, presence of active bleeding, and need for surgery. Logistic regression analysis was used to identify significant clinical determinants of transfusion. Red blood cell transfusion rate was 8%. Of admission diagnoses and co-morbidities, only cancer was associated with a trend for transfusion (p=0.07). Those who received red blood cell transfusion had significantly lower admission hemoglobin levels (p=0.005), higher serum creatinine (p=0.02), and lower mean arterial pressure (p=0.01). Significant predictive clinical factors of red blood cell transfusion included congestive heart failure (OR=3.72, CI 1.034, 27.160), admission hemoglobin level (d/L, OR=0.956, CI 0.927, 0.986), mean arterial pressure (mm Hg, OR=0.942, p=0.005) and serum creatinine (ml/min, OR=1.008, CI 1.003, 1.014). In conclusion, predictors of red blood cell transfusion for medically ill patients include low admission hemoglobin, renal insufficiency, and low blood pressure.
The Temporal Gene Expression Patterns of Embryonic Hematopoiesis after Hemangioblast Commitment.
Authors: Chen Dong , Lewis Rachel L. , Daigh Christine A. , Wang Ping , Kendzioski Christina M. , Chen Xin , Brown Patrick O. , Thomson James A. 
Published: April 2020

Abstract
Human embryonic stem (ES) cells provide a unique model for studying the development and function of human tissues. To better understand early development of the hematopoietic lineage, here we use microarray expression analysis to examine the temporal patterns of gene expression in embryoid bodies derived from human ES cells around the time when the definitive hematopoiesis peaks. We use an empirical Bayes hierarchical modeling approach, called EBarrays, to assess each of the possible temporal patterns of gene expression for the 5 time points examined, and correlate those patterns with the emergence of definitive hematopoiesis. We find that several genes known to be associated with early hematopoietic commitment, such as Flk-1 and TAL-1/SCL, are expressed 4-6 days before the peak colony formation cells (CFCs) production. The expression of a distinct group of genes previously identified as important in adult hematopoietic self-renewal (such as PIK3R1, ABCB1/MDR-1, RGS18, IRS1, SENP6/SUMO-1, and Wnt5A etc. correlated with the emergence of definitive hematopoiesis. Novel genes demonstrating the same expression pattern as this group are likely to have critical roles in the development and self-renewal of definitive hematopoietic precursors.
Cocultures of Umbilical Cord Blood Hematopoietic Progenitor Cells and Umbilical Cord Endothelium.
Authors: Moldenhauer Anja , Selders Lisa , Genter Gesche , Hieronymus T. , Ruau D. , Lun Andreas , Futschik Matthias , Kiesewetter Holger , Zenke Martin , Salama Abdulgabar 
Published: April 2020

Abstract
Purpose. The expansion of hematopoietic progenitor cells (HPC) from cord blood is limited so far. For HPC expansion and differentiation we evaluated a coculture model of cord blood-derived HPC and endothelial cells (EC) isolated from the umbilical cord.
Methods. CD34(+) HPC were cultured directly in contact with the endothelial monolayer or indirectly using a 0.4μm microporous transmembrane. EC were stimulated by various cytokines including TPO, interleukin (IL) 1β, IL4, and IL6. After 1 and 2 weeks, the HPC were counted and analyzed by flow cytometry. Their plating efficiencies were determined by methylcellulose assays, and their reconstitutive potential was analyzed by cobblestone assays. DNA microarrays on the expanding cells as well as on cytokine-stimulated EC were also performed.
Results. EC stimulation with thrombopoietin and IL4 had no significant effect on mononuclear cell expansion. In contrast, IL1β stimulation resulted in a 10 and 100fold increase of the cell number after one and two weeks, respectively. More than 90% of these cells were CD33(+) and had a plating efficiency equivalent to that of HPC post isolation (7.3 ±1.2 versus 8.3±0.68%). The number of 5-wk cobblestone-area forming cells among these expanded cells was diminished, whereas the number of 2-wk cobblestone areas did not change (10.6 versus 8.5). The expanded cells could further mature into functional granulocytes. Time-course DNA microarrays of IL1β-stimulated EC demonstrated an increased expression of genes encoding tumor necrosis factor-induced proteins, interleukins, and endothelial adhesion molecules. When IL1β was replaced by IL6, a 7-fold expansion of morphologically progenitor-like cells took place. 10–13% of these cells were positive for CD133, had a plating efficiency of 10.2±1.7%, and showed on average 14.5 five-week cobblestone areas.
Conclusion. Cocultures of cord blood-derived HPC with umbilical cord-derived EC offer alternative methods for the expansion and differentiation of hematopoietic cells.
Granulocyte-Colony Stimulating Factor Decreases Angina and Improves Exercise Capacity in Patients with Chronic Refractory ‘No Option’ Ischemic Heart Disease.
Authors: Ma David D.F. , Kovacic Jason C. , Dodds Anthony , Tao Helen , McDonald Peter , Feneley Michael P. , Muller David W. , Fruend Judith , Milliken Sam , Moore John , Itescu Silviu , Graham Robert M. 
Published: April 2020

Abstract
Granulocyte-Colony stimulating factor (G-CSF) is widely used in the treatment of hematological malignancies. The administration of G-CSF is a novel therapy for ischemic heart disease (IHD), but current data are controversial and the safety and efficacy of G-CSF in acute or chronic IHD is unclear.
Aim: to assess safety and efficacy of G-CSF administration and stem cell mobilization in patients with chronic refractory ‘no option’ IHD.
Methods: After baseline cardiac assessment (CA) [Seattle Angina Questionnaire (SAQ), Bruce exercise stress test (EST), persantin-Sestamibi and dobutamine-echocardiographic imaging], stable ‘no option’ IHD patients received open-label G-CSF 10μg/kg for 5 days, with an EST (to facilitate myocardial cytokine generation and stem cell trafficking) on the 4th and 6th days. After 3 months, CA and the same regimen of G-CSF and ESTs was repeated, but in addition, leucopheresis and a randomized double-blinded intracoronary infusion of either CD133+ or unselected cells was performed (randomized data remains blinded). Final CA was 3 months thereafter.
Results: Thirteen patients (12 male, 1 female, mean age 62) received 21 cycles of G-CSF. There were no deaths, Q-wave AMIs or any complications with long-term sequelae, although, transient troponin-I elevation (n = 3) and thrombocytopenia (n = 2) were observed. Mean CD133+ cell count rose from 1.28 to 56.12 x1012/L (p = 0.001). There was no age-related trend towards lower numbers of G-CSF mobilized CD34+ cells in the IHD patients, as compared to a group of younger (&lt;60 y.o) normal male donors (n = 66). After the first cycle of G-CSF, SAQ angina frequency and physical limitation scores improved, and EST time increased (all p &lt; 0.01). There was further but less marked improvement in each of these parameters after the second cycle of G-CSF and cell infusion. Overall, SAQ angina frequency score improved 46 points (95% CI +22 to +70, p = 0.003). This was reflected by reduced anginal frequency and nitrate use (both p &lt; 0.005). Overall, SAQ physical limitation score improved 26 points (95% CI +17 to +35, p = 0.0003), EST time improved 97 seconds (95% CI +41 to +153, p = 0.005) and Duke Treadmill Score improved 4.3 points (95% CI −0.2 to +8.8, p = 0.058).
Conclusions: G-CSF and intracoronary cell infusion is safe in chronic ‘no option’ IHD patients. In this phase I study G-CSF improved anginal frequency, nitrate use and EST performance. A placebo controlled phase II trial investigating G-CSF with an appropriate cytokine stimulus is warranted.
Involvement of Lipid Raft Domain Signaling in Ex Vivo Maintenance of Hematopoietic Progenitor Cells.
Authors: Wirthlin Louisa , Meyerrose Todd E. , Creer Michael H. , Nolta Jan A. 
Published: April 2020

Abstract
Ex vivo manipulation of hematopoietic stem cell populations has been well documented to result in loss of pluripotentiality, proliferative deficits, and a decreased ability to repopulate post-transplantation. These defects result from a multitude of factors, and here we examine the contribution of lipid raft-localized signaling complexes and pro-survival growth factors, such as hepatocyte growth factor (HGF), in mediation of these defects. Specifically, we have examined the effects of diffused oxygen and hepatocyte growth factor (HGF) on ex vivo culture conditions for preservation of hematopoietic stem cell potentiality. We hypothesize that the high level of diffused oxygen in traditional cell culture systems represents a cellular stress leading to differentiation and loss of potential. We further hypothesize that this effect can be mitigated by the pro-survival factor HGF. Human umbilical cord blood derived mononuclear cells were first isolated by a ficoll density gradient followed by immunomagnetic depletion of mature cell lineages. These lineage depleted cells (Lin-) were cultured on fibronectin-coated plates in serum free media and kept in a 37° humidified atmosphere. We supplemented the cultures with stem cell factor (SCF), Flt-3, thrombopoietin (TPO), and HGF, and then manipulated the available oxygen for the cultured cells between 18% (ambient) and 2–3% (hypoxic). The cells were harvested at various time points (24h, 48h, 72h, and 96h) and assayed for lineage cell markers in conjunction with surrogate markers of lipid raft formation by flow cytometry. Preservation of phenotypic cell surface markers such as CD34, CD133, and CD31 were assayed, as well as the presentation of c-met, the receptor for HGF. Results were validated by immunohistochemistry and confocal microscopy for colocalization of receptor-raft complexes. We found that total lipid raft expression was upregulated in hypoxic conditions versus ambient, with a significant increase in a population of cells that highly expressed lipid raft markers. In addition, we found that cells cultured in the presence of HGF in ambient conditions showed a decrease in the presence of lipid rafts with a trend toward lower expression in those cells cultured with HGF. This population of cells demonstrated a specific preservation of CD34+ expressing cells, and decreased incidence of apoptosis in all compartment surveyed. Functional assays of colony-forming ability, SCID-repopulating activity, and competitive repopulation were also assayed, to correlate the upregulation of lipid raft domain presentation with relevant stem cell ability. Current focus of this investigation centers on the identification of the specific receptor signaling complexes associated with these raft domains.
Isolation and Characterization of Murine Novel cDNA Clone Using Newly Constructed Gene Trap Vector.
Authors: Shirasaki Ryosuke , Tashiro Haruko , Noguchi Mitsuho , Goto Moritaka , Kawasugi Kazuo , Shirafuji Naoki 
Published: April 2020

Abstract
Objective: To understand better the mechanism of the maintenance of immature state of stem cells, murine novel cDNA clones are to be isolated by the gene trap method.
Method: The trap vector was constructed with En-2 intron sequence followed by splicing acceptor sequence, IRES signal sequence and b-galactosidase cDNA. NeoR gene was also contained without poly-A additional signal. After electroporation into Embryonic stem (Es) cell lines (D3, and E14.1 Köln), neomycin-resistant Es clones were picked up, and were selected in which the expression of b-gal was detected in the undifferentiated condition cultured with leukemia inhibitory factor (LIF), and feeder layer cells by X-gal staining, but were not detected in the differentiated condition with ATRA, and without LIF nor feeder layers. Using 5′ RACE method, the transcripts from the selected clones were identified. One clone was further analyzed. Full-length cDNA was isolated from a library constructed from Es D3 cells, and its characterization was determined.
Result and Discussion: one unique full-length cDNA clone was isolated for the further characterization. It was constructed with 2225 nucleotides, and putative 466 amino acids were encoded in a single long open reading frame. The transcript was detected in the undifferentiated Es cells but not in differentiated Es cells nor in various kinds of organs by northern blotting analysis. When this clone was over-expressed in murine 10T1/2 cells, RT-PCR analysis demonstrated the down-regulation of Id-1 gene. These data suggested that the isolated cDNA clone was possibly related to keeping Es cells to be immature multipotent state. We now try to isolate the related cDNA clones which are expressed mainly in the hematopoietic stem cells.