bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Risk of Venous Thrombosis among Women with Thrombophilia and Oral Contraceptive Use: A Meta-Analysis
Authors: Dayan Natalie , Tagalakis Vicky , Holcroft Christina 
Published: April 2020

Abstract
Introduction: Several small studies have reported an increased risk of venous thrombosis (VT) with thrombophilic disorders that is compounded by the use of oral contraceptives (OCP). The objective of this meta-analysis was to summarize the risk of VT among women with thrombophilia and OCP use, and to assess the interaction between OCP use and thrombophilia.
Methods: Data sources used included MEDLINE, EMBASE, CINAHL, and COCHRANE REVIEW databases (1950–2008), and reference lists of retrieved articles. We selected studies that assessed among reproductive-aged women with and without VT the prevalence of OCP use and thrombophilic disorders, including factor V Leiden (FVL), prothrombin (PT) G20210A, elevated procoagulant factor VIII, hyperhomocysteinemia, lupus anticoagulant, and deficiencies of proteins C, S and antithrombin III. VT was defined as objectively confirmed deep vein thrombosis, pulmonary embolism or cerebral vein thrombosis. Fifteen studies were included. Two independent reviewers extracted data on study characteristics, quality, risk factors, and outcomes. Odds ratios (ORs) were calculated for each study and pooled using the random effects model.
Results: There was an increased risk of VT in women with OCP use (pooled OR 3.0, 95% confidence interval (CI) 2.0–4.5), women with any thrombophilic disorder (pooled OR 4.8, 95% CI 3.6–6.5), women with FVL (pooled OR 4.7, 95% CI 3.1–7.5), and women with PT G20210A (pooled OR 3.3, 95% CI 0.8, 13.1). OCP users with FVL had a 18-fold increased risk of VT compared to healthy OCP non-users (pooled OR 17.8, 95% CI 5.6–56.8) and OCP-using women with PT G20210A had a 12-fold increased risk (pooled OR 12.4, 96% CI 0.9– 166) compared to healthy OCP non-users. The association of OCP use and VT was of similar magnitude among women with FVL (pooled OR 3.2, 95% CI 1.3–7.7) and women without FVL (pooled OR 3.9, 95% CI 1.8 to 8.6). Significant heterogeneity (I2 &gt;80%) was present among models for OCP use, PT G20210A, and OCP use among FVL-negative women.
Conclusions: We have shown that OCP use, thrombophilia, and in particular FVL and PT G20210A are associated with an elevated risk of VT of similar magnitude. However, the combination of OCP use and thrombophilia further amplifies the risk of VT compared to when either of the risk factors are considered alone. Though heterogeneity was observed, our study suggests, and confirms previous reports of an interaction between OCP use and thrombophilia.
Prevalence of Thrombophilia in Patients with Retinal Vascular Occlusion
Authors: De Sancho Maria Teresa , Powell-Boone Karen , Carlson Karen 
Published: April 2020

Abstract
Background: Retinal vascular occlusion (RVO), both arterial (RAO) and venous (RVO) is an important cause of visual loss. Inherited and acquired thrombophilia are considered important risk factors in the development of RVO. Additionally, cardiovascular risk factors such as hypertension, diabetes, hyperlipidemia, smoking and use of female hormones may play a role.
Objective: To determine the prevalence of thrombophilia in patients with RVO.
Methods: We reviewed the medical records of all patients with documented RVO who were referred to our hematology clinic at a tertiary care center between January 2001 and June 2008 for thrombophilia evaluation. Thrombophilia evaluation included testing for factor V Leiden, prothrombin gene mutation G20210A, deficiencies in antithrombin, protein C and S, presence of antiphospholipid antibodies (aPLas), hyperhomocysteinemia and increased levels of lipoprotein (a), factor VIII, IX and XI. Serum protein electrophoresis and immunofixation were also performed. Environmental risk factors analyzed included use of female hormones, pregnancy and puerperium, smoking, hypertension, hyperlipidemia and diabetes.
Results: A total of 24 patients were identified; 10 were males and 14 females. The mean age was 62 years (range, 31 to 86 years). Seven patients had RAO and 17 RVO. Of the 24 patients, 19 (79%) had thrombophilia. Of these, 10 had only one thrombophilia and 9 had ≥2 thrombophilia. Of the 10 patients with a single thrombophilia, 3 patients had aPLas, 3 had increased lipoprotein (a) 2 had factor V Leiden, 1 hyperhomocysteinemia, and 1 had increased factor VIII level. Of the nine patients with combined thrombophilia: 3 had hyperhomocysteinemia and increase factor VIII level, 2 prothrombin gene mutation and positive lupus anticoagulant, 1 positive aPLas, hyperhomocysteinemia and a monoclonal gammopathy, 1 aPLas and increased factor VIII, 1, hyperhomocysteinemia and protein S deficiency and 1 hyperhomocysteinemia and polycythemia vera. In 5 patients, no thrombophilia was identified. Of the 24 patients, 11 had hypertension, 9 had hyperlipidemia, 3 were smokers, 3 were using female hormones and 1 was pregnant. Treatment included aspirin 81 mg (n= 9), aspirin &gt;81 mg (n= 5), warfarin (n= 3), folic acid (n=2), aspirin and clopidogrel (n=1), clopidogrel (n=1), no treatment (n=4), and unknown (n=1).
Conclusions: Over three quarters (79%) of patients with RVO had an underlying thrombophilia with aPLas, hyperhomocysteinemia and increased factor VIII level being the most prevalent. Our findings suggest that testing for thrombophilia particularly for aPLas, homocysteine and factor VIII levels may be useful in patients with RVO
A Rare Cause of Anemia in Inflammatory Bowel Diseases: A Case Report and Review of Literature
Authors: Ahmed Waqas , Monroe Kevin , Essell James , Broun E. Randolph 
Published: April 2020

Abstract
Introduction: Anemia is a common problem in patients with inflammatory bowel diseases (IBD), and its etiology is usually multifactorial. It can be produced by chronic blood loss, nutritional deficiencies, and drugs such as salazopyrine; however it can also due to auto immune hemolysis, which is a rare complication of IBD. We report a case of coombs positive autoimmune hemolytic anemia associated with ulcerative colitis both diagnosed at the same presentation.
Case Report: A 32 year old man with no significant past medical history presented with complaint of dark colored urine, jaundiced skin and fatigue for 4 weeks. He also reported diarrhea mixed intermittently with blood for last few months. Physical exam was consistent with jaundice and anemia (pallor and icterus) with slightly palpable spleen. Initial lab work up showed Hb of 3.8 with normal platelet and WBC count, high reticulocytes count of 7% .LFT showed serum bilirubin of 3.6 (direct 0.4) with normal serum ALT and AST levels .Serum LDH was high (1032 U/l) while serum haptoglobin was low (0.11 mg/dl). Peripheral smear showed anisopoikilocytosis & spherocytosis. (See Figure 1) Further investigations revealed a positive direct Coombs test consistent with diagnosis of autoimmune hemolytic anemia. CT abdomen and pelvis showed mild splenomegaly & non-specific enlarged mesenteric lymph nodes. Colonoscopy revealed ulcerative pancolitis confirmed by histological findings of biopsies taken. Patient received PRBC transfusions and was started on steroids and mesalamine and was discharged on maintenance dose. His symptoms resolved in 4 weeks and Hb remained stable with no evidence of further hemolysis at 4 month follow up .Repeated CT abdomen & pelvis showed resolution of the lymphadenopathy.
Figure 1: Peripheral Blood smear showing anisopoikilocytosis & spherocytosis. Figure 1:. Peripheral Blood smear showing anisopoikilocytosis & spherocytosis.
Discussion: Autoimmune hemolytic anemia (AIHA) is a rare complication of IBD. The exact underlying pathogenesis of this association remains obscure; however it has been attributed to the production of cross reacting anti erythrocyte antibodies. In AIHA associated with IBD, corticosteroids are considered to be first line therapy and often cause remission of hemolysis along with treatment for IBD Immunomodulators and splenectomy has been used for patients with refractory AIHA. Colectomy done for fulminant colitis has also been reported to induce remission of AIHA. Further studies for long term follow up and pathogenesis of this association are warranted.
Identification of Novel Phosphorylation Sites and Kinetics in Flt3 Receptor Wt, ITD and D835Y
Authors: Razumovskaya Elena , Masson Kristina , Khan Rasheed , Bengtsson Susanne , Ronnstrand Lars 
Published: April 2020

Abstract
FMS-like tyrosine kinase-3 (Flt3) is a receptor tyrosine kinase, which is normally expressed in hematopoietic progenitor cells. It has been implicated as a major cause of transformation in acute myeloid leukemia (AML). There are two types of Flt3 gene mutations have been identified in AML: duplication of amino acids in the juxtamembrane region- Internal Tandem Duplication (ITD) and an activation loop point-mutation of D835 in kinase domain. These mutations cause constitutive activation or over expression of Flt3 receptor and therefore lead to alteration in signal transduction. These alterations occur in approximately 30% of AML patients. High occurrence of these mutations in the Flt3 receptor in AML patients makes it one of the most interesting therapeutic targets. In this study we have identified three novel in vivo phosphorylation sites of Flt3 receptor and further compared the activity of phosphorylation sites of Flt3 wild type, Flt3 ITD and D835Y mutations by using homemade phospho-specific antibodies directed against specific tyrosines. For this study murine hematopoietic Ba/F3 cells were stably transfected with wild-type Flt3, ITD and D835Y mutations. We have confirmed that the activation of the wild type Flt3 receptor is ligand dependent and response in a time dependent manner, but Flt3-ITD and D835Y are constitutive active and ligand independent. Phosphorylated tyrosines 589, 591, 599, 726, 768, 793, 842, and 955 of Flt3 receptor were investigated and shown to be differentially activated in wild-type versus the mutated receptor. Using this data we can further study the mechanisms of signaling pathways of the Flt3 receptor that are involved in many biological responses and understand the mechanism by which Flt3 ITD and D835Y functions in pathological conditions.
Generation of Endothelial Progenitor Cells from Amniotic Fluid Stem Cells
Authors: Cardier Jose E. , Wittig Olga , Alonso Jose , Romano Egidio 
Published: April 2020

Abstract
Recently it has been shown that human amniotic fluid contains stem cells of fetal origin (AFS) that express embryonic and adult stem cell markers. AFS can be expanded extensively in culture and give rise to multiple differentiated cells such those of myogenic, osteogenic, neurogenic and endothelial lineages, all of them with potential therapeutic value. In this study, we investigated the generation of endothelial progenitor cells (EPC) from AFS. For this purpose, remnant amniotic fluid was obtained from mothers undergoing amniocentesis for medical reasons at 18 weeks of pregnancy. Cells were collected by centrifugation and cultured in DMEM, supplemented with Chang medium and fetal bovine serum. Adherent cells were expanded and characterized by flow cytometry at various times using antibodies for CD34, CD45, CD133, CD117, KDR (VEGFR-2) and CD31. Cultures were also carried out in endothelial growth medium and analyzed for EPC markers (CD133, KDR, CD31). Adherent cells showed, at 8 and 17 days of culture, fibroblastoid features. Flow cytometry analysis, at 2 weeks of culture, showed cells expressing CD 133 (37%), CD 34 (1%), CD 117 (3%) and KDR (12%). Cultures carried out in endothelial differentiation medium (EDM), at 33 days of culture, showed a significant increase of cells expressing EPC markers: CD133 (40%), CD34 (3%) KDR (24%), KDR/CD31 (9%). These cells formed blood vessel-like structures on collagen-coated plates. The in vivo angiogenic capacity of these cells was demonstrated by inoculating them into collagen microspheres and implanting them subcutaneously into immunocompromised mice. Histologic examination revealed that these cells formed microvessels on implantation. Our data indicates that EPC can be generated and expanded from AFS, and they might be useful for therapeutic angiogenesis in ischemic tissues.
Changes in MRI-Estimated Cardiac and Hepatic Iron Load in β-Thalassemia Patients Treated with Deferasirox at a Single Institution
Authors: Kattamis Antonios , Moraitis Panayiotis , Lourida Athanasia , Rigatou Efthemia , Ladis Vassilios 
Published: April 2020

Abstract
Chronic transfusions have dramatically improved the life expectancy of patients with β-thalassemia, but leads to severe iron overload. Long-term iron chelation therapy aims both in the prevention and the reversal of iron load and iron toxicity. The available chelators seem to have different profiles of relative heart and liver iron removal. Data on cardiac efficacy of deferasirox (DFX; Exjade®) are limited. We report results on patients with thalassemia, followed in a single institution, who completed at least 12 months of treatment with deferasirox.
Methods: Fifty five patients (22 males, 33 females; mean age: 27±7.1 years, splenectomised: 5 patients, HCV seropositive: 7 patients) were included in the study. The patients were treated with deferasirox at a mean dose of 27.2±5.2 mg/kg/day (range 15–40 mg/kg/day). Sixty-one pairs of MRI assessments of cardiac and hepatic iron overload by using the T2* technique were performed at a mean time interval of 13.7±2.4 months. Cardiac function parameters, mainly ejection function (EF), were also estimated at the same time.
Results: Mean, SD and range at the time of the first MRI evaluation showed: ferritin: 2520±1584 ng/mL (371–6953 ng/mL), cardiac T2*: 25.3±10.7 ms (3.8–39.2ms), liver T2*: 4.5±4.6 ms (0.57–21.1ms) and ejection fraction: 66.9±4.8 ms (55.5–75.4%). Follow up assessments showed: ferritin: 2375±1743 ng/mL (39–7245 ng/mL), cardiac T2*: 27,1±11.7 ms (2.7–40.8ms), liver T2*: 6.2±6.2 ms (0.5–26.4ms) and ejection fraction: 66.1±4.8 ms (55.8–75.4%). Changes in liver and cardiac T2* were statistically significant (p=0.0004 and 0.008, respectively). Changes of the cardiac T2* values correlated well with the dose of deferasirox, while changes of liver T2* with ferritin changes (r=−0.284, p=0.032, r=−0. 27 and p=0.046, respectively). A statistical significant improvement (p&lt;0.05) in cardiac and hepatic iron load was observed in the subgroup of 22 patients with severe liver iron load (defined as hepatic T2*&lt;1.9 ms). In 18 patients with moderate/severe cardiac iron load (cardiac T2*&lt;20ms), T2* decreased in 5, remained stable in 2 and improved in 11 patients (mean 10.7±5.5 ms vs. 12.1±7.1, p=0.078).[S1]
Conclusions: This observational study shows that treatment with deferasirox improves both hepatic and cardiac iron overload in regularly transfused β-thalassemia patients. Adjustment of the dose is required to enhance iron excretion and achieve better negative iron balance
Peptide Vaccine Preparation and Validation with a Bio-Assay
Authors: Schmitt Anita , Bechter Clemens , Yao Junxia , Goetz Marlies , Maccari Birgit , Schauwecker Peter , Wiesneth Markus , Schmitt Michael 
Published: April 2020

Abstract
Introduction: Peptide vaccination constitutes a novel immunotherapeutical approach for the treatment of patients with solid tumors, lymphoma and leukemia. Moreover it might be of use in hematooncological patients for the prevention and therapy of infections like cytomegalovirus (CMV) reactivation due to immunosuppression. To meet good manufacturing practice (GMP) criteria, we introduce here a bio-assay to validate peptide vaccines for peptide content and bio-activity.
Methods: As a paradigm for peptide vaccine preparation the immunogenic CMV peptide (pp65, pos.: 495–503 NLVPMVATV) lyophilisate was resolubilized in dimethyl sulfoxide (DMSO), phosphate buffered saline (PBS) and admixed with Montanide™ (ISA51: incomplete Freund’s adjuvant [IFA]). This mixture freshly prepared, stored at +4°C and cryopreserved at −20°C was subjected to lymphocyte peptide cultures (MLPC) followed by enzyme linked immunospot (ELISPOT) assays and flow cytometry (FACS) as bio-assay.
Results: Addition of different amounts of peptide (10–80 μg) to a MLPC resulted in the generation of interferon (IFN) gamma and granzyme B releasing CD8+ CMV tetramer+ T cells in a dose dependent manner. The combination of FACS and ELISPOT results allowed the definition of the peptide amount in a vaccine preparation. Storage at ±4°C over 24 hrs did not result in a significant change of the immunogenicity of the vaccine, as assessed by ELISPOT and FACS. In contrast, cryopreservation of the vaccine at −20°C resulted in a loss of immunogenicity.
Conclusion: Quantitation of tumor/viral antigen peptides admixed with adjuvants, such as IFA, is feasible through bio-assays as the modified ELISPOT/FACS assay described here, meeting GMP criteria for multi-center trials.
Endothelial Progenitor Cell Marker CD133 Identifies Tumor Endothelium
Authors: Baljevic Muhamed , Shmelkov Sergey V , Nolan Daniel J , Hooper Andrea T , Hormigo Adilia , Iyer Rajiv , Gutin Philip H , Rafii Shahin 
Published: April 2020

Abstract
CD133, a pentaspan surface antigen, is considered a marker of tissue-specific stem cells, including hematopoietic stem cells and endothelial progenitors. We and others have previously shown that CD133 expression is high on endothelial progenitor cells; that it is downregulated during their differentiation and that it is not found on mature endothelium. In addition, CD133+ cells in brain tumors were shown to exhibit features of cancer stem cells. Recent report showed that CD133+ tumor cells are located in perivascular niches in astrocytomas and glioblastomas. It was also demonstrated that in glioblastomas, CD133+ cancer stem cells are tightly associated with tumor vasculature and that signals from endothelial cells are the key factors in self-renewal and proliferation of cancer stem cells. However, the contribution of CD133+ endothelial progenitor cells to tumor angiogenesis remains unknown. We sought to investigate the role of CD133+ cells in human brain tumors and their relation to endothelial progenitor cells and to the mature endothelium. To this end we examined tumor samples resected from patients with glioblastomas. Histological and immunohistochemical analyses revealed that CD133+ cells in these tumors are predominantly endothelial cells as demonstrated by co-staining with CD31 and CD133. We further analyzed a population of endothelial cells using multicolor flow cytometry analysis and we demonstrated that on average 60% of endothelial cells, as defined by the expression of CD31 and VE-cadherin, also express CD133. However, we also found significant heterogeneity between glioblastomas obtained from different patients. Additionally, CD133 was not expressed on CD45−CD31+ subpopulation in meningiomas. Taken together, we demonstrated that high percentage of endothelial cells in glioblastomas expresses CD133, while CD133 could also be found on other cell types within the tumor. Furthermore, our data suggest that tumor vasculature might be enriched with bone marrow derived hemangiogenic progenitors.
Human Bone Marrow Mononuclear Cells, CD34+ Cells and CD133+ Cells Do Not Form Vascular Structures in Vivo
Authors: Josefsen Dag , Kuchler Axel , Wang Mengyu , Haraldsen Guttorm , Kvalheim Gunnar 
Published: April 2020

Abstract
The clinical results of using bone marrow-derived progenitor cells to repair ischemic tissue in coronary disease are controversial since some studies demonstrate improvement of cardiac function, whereas others do not. In these studies both unmanipulated mononuclear bone marrow cells (MNC) and highly enriched stem cells such as CD34+, CD133+ or mesenchymal cells (MSC) were used. In this study we have examined the in vitro and in vivo endothelial progenitor properties of these populations. MNC were obtained from whole BM. CD34+ cells and CD133+ cells were isolated using magnetic bead technology (MACS, Miltenyi). MSC was established from BM, and only second passages from MSC were used. Cytokine profiles of MSC were measured on a Bio-Plex system (BIO-RAD Laboratories). In vitro studies were performed by culturing cells in either complete EGM-2 medium to identify EPC, or in M199 with endothelial growth factors to investigate endothelial differentiation. We observed little or no endothelial differentiation when culturing CD133+ or CD34+ cells in M199 media. Furthermore, we did not observe EPC colony formation of MNC, CD133+ cells, or CD34+ cells cultured in EGM-2 medium. To study the in vivo effect of MNC, CD34+, CD133+ as well as MSC, we adoptively transferred human cells into immunodeficient mice and examined their endothelial potential assessing the formation of blood vessels as readout (Skovseth et al Am J of Pathol 2002, Blood 2005). Human umbilical cord-derived endothelial cells (HUVEC) served as positive controls. MNC only grew as round shaped clustered cells with no signs of vessel formation. Neither CD34+ cells nor CD133+ cells demonstrated signs of endothelial development in vivo. MSCs were found to ex vivo secrete endothelial growth factors, including VEGF, bFGF and EGF. When Matrigel plugs with MSCs were examined, an accumulation of smooth muscle actin-positive cells could be detected, but no vessel like endothelial structures derived from human cells could be found. If these putative smooth muscle cells are differentiated from the MSCs, or are murine host cells attracted to the human MSCs due to the cytokine secretion effects of the MSCs is not clear. Currently, we are exploring if co-cultivation of MSC in combination with MNC, CD133+ cells or CD34+ cells will facilitate endothelial development. In conclusion, our experiments did not show endothelial cell differentiation or vessel formation from human BM-MNC, CD34+ cells or CD133+ cells suggesting that such cells have a limited vasculogenic effect. Our findings with the MSCs are interesting and the mechanism of actions is under further investigation.