bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative Analysis of Gene Expression for Vascular Endothelial Growth Factor and Its Application.
Authors: Bai Xia , Fu Jianxin , Ding Kaiyang , Cen Jiannong , Wang Zhaoyue , Ruan Changgeng 
Published: April 2020

Abstract
Vascular endothelial growth factor (VEGF) is a crucial mediator of angiogenesis, and plays an important role in pathogenesis of leukemia. However, the importance of VEGF in differentiation or apoptosis of leukemic cells remains to be evaluated. A competitor DNA fragment template of VEGF gene mimic was constructed with the method of gene recombinant technologies, and a competitive quantitative reverse transcriptase-polymerase chain reaction (cQRT-PCR) for analyzing VEGF gene expression was performed to assess the regulation of VEGF gene expression in the process of all-trans retinoic acid (ATRA)-induced differentiation of an acute promyelocytic leukemia cell line NB4. In construction of a standard curve from which the amount of target cDNA was derived, serial dilutions of the target were co-amplified with a constant amount of mimic, and the intensities of bands corresponding to the target and the mimic were measured. CD11b antigen and nitroblue tetrazolium (NBT) reduction rate of NB4 cells were also assayed at different time points. cQRT-PCR was a sensitive and reliable tool for analysis of VEGF gene expression, with a detectable range from 1 to 2 times 10 the fifth power molecules. The number of VEGF gene transcripts detected by means of cQRT-PCR assay was 42.3, 12.6, 3.6, and less than 1.0 times 10 the fifth powder per microgram of NB4 total RNA at 0, 12, 24 and 48 hours after ATRA treatment, respectively. The rapid down-regulation of VEGF gene expression during ATRA-induced NB4 cell differentiation was accompanied by an upregulation of CD11b expression and an increased NBT reduction rate. In conclusion, cQRT-PCR could be used as an efficient method of qualitative analysis of VEGF gene expression. ATRA significantly depresses VEGF expression and its antileukemic effect can be brought through the two ways of differentiation induction and angiogenesis inhibition.
Pharmacogenomics Study of Biological Pathways Related to Bortezomib Activity in Myeloma.
Authors: Mulligan George , Bryant Barb , Roels Steven , Bolt Andrew , Meng Yan , Koenig Erik , Boral Anthony , Esseltine Dixie-Lee , Anderson Kenneth A. , Bergsagel Peter Leif , Sonneveld Pieter , Broyl Annemiek , Mitsiades Constantine , Chng Wee-Joo , Richardson Paul G. , Trepicchio William 
Published: April 2020

Abstract
Introduction: Clinical development of bortezomib in multiple myeloma (MM) included pharmacogenomic gene expression profiling (GEP) of pre-treatment tumor biopsies in order to evaluate the feasibility of developing predictive classifiers in prospective clinical trials and to better define the biological pathways related to drug sensitivity. By partitioning the samples into train (n=101) and test (n=80) datasets, statistically significant classifiers of response and survival were identified and then validated [Mulligan G, et al: Blood. 2007; 109:3177–88]. Here we describe specific genes and biological pathways associated with bortezomib efficacy in relapsed MM.
Methods: Using tumor GEP data from 188 patients treated with single-agent bortezomib we identified genes associated with overall survival (OS), as well as genes associated with response (R) or progressive disease (PD) after bortezomib therapy. These clinical endpoints were also analyzed with biological pathway algorithms including GSEA [Subramanian A, et al: Proc Natl Acad Sci2005; 102:15545–50] Ingenuity Pathway Analysis and Gene Ontology/GATHER. These results were then compared to GEP data from a patient subset (n=76) treated with single-agent dexamethasone and also cell lines selected for differential sensitivity to bortezomib.
Results: A comparison of R and PD highlighted numerous differentially expressed genes; approximately 200 were over-expressed in R and 500 were over-expressed in PD (t-test &lt;0.01). R-associated genes include signaling molecules like TRADD, CFLAR and TANK, while PD-associated genes included oncogenes (NRAS), apoptotic regulators (DAP3) and cancer antigens (CTAG). Pathway tools helped to integrate and extend these observations, consistently identifying NFkB activation and overexpression of adhesion pathways as R-associated. Overexpression of protein synthesis and mitochondrial pathways were PD-associated. The overexpression of pathways such as NFkB, adhesion and cytokines appear linked specifically with bortezomib and not dexamethasone sensitivity. These approaches were also applied to patient survival. In contrast to analyses of single genes, the pathway tools highlighted links between response and survival, including adhesion and inflammatory cytokine pathway expression.
Conclusions: This study highlights the diversity of biological pathways expressed in MM and the utility of clinical genomics to associate pathways with activity of single-agent bortezomib.
Thrombotic Thrombocytopenic Purpura with Normal Initial Hemoglobin.
Authors: Yarlagadda Madhavi , LePar Felice 
Published: April 2020

Abstract
Thrombotic thrombocytopenic purpura (TTP) is a life threatening multisystem disorder considered a true medical emergency. Usually patients are said to present with low hemoglobin as well as low platelets. This is a case of patient with early TTP having normal hemoglobin at presentation. A 37 year old male with history of idiopathic thrombocytopenic purpura (ITP) about 20 years ago, Ulcerative Colitis s/p colon resection, Graves disease s/p ablation, Lupus anticoagulant, and TTP about 3 years prior to admission which was treated with plasma exchange, was in his usual state of health until the morning of admission when he developed red urine and petechiae on his shoulder after playing football with his son the day before. He also reported a small amount of blood when he blew his nose. He denied fevers, chills, headache, confusion, anuria or any recent viral illness. Physical exam on admission revealed stable vital signs without fever, mild scleral icterus, and diffuse petechiae on shoulders/chest and ankle. He was alert and oriented × 3. Lab data showed platelets 5 k/ul, hemoglobin 14.3 g/dl, white blood cells 7.7 k/ul, BUN 29 mg/dl, creatinine 1.0 mg/dl, total bilirubin 6.6 mg/dl, direct bilirubin 0.5 mg/dl, reticulocytes 1.3%, LDH 1245 u/l, and haptoglobin &lt; 7 mg/dl. Direct Coombs was negative. Peripheral smear showed 5–6 schistocytes/HPF. DIC work up was negative. Urinalysis showed large occult blood, 83 RBCs and 300mg/dl of protein. We suspected an early presentation of recurrent TTP. He was treated with methylprednisolone 1mg/kg and FFP 2–3 units then therapeutic plasma exchange with initial good response. His hemoglobin dropped to 10.9 within the next 36 hrs. He required plasma exchange over 3 months as well as other treatments including steroids, vincristine, splenectomy, and rituximab. Eventually he remained in remission. This case is notable for the normal initial hemoglobin. Isolated thrombocytopenia is often said to be ITP, but we believe that in this case the patient presented so soon after the development of symptoms that the hemoglobin had not yet declined. As TTP can be fatal if not recognized and treated promptly, it may be prudent to evaluate all patients with acute profound thrombocytopenia for TTP.
Hemophilia Treatment - A Model for Appropriate Cost Control.
Authors: Aledort Louis M. , Coates James G. 
Published: April 2020

Abstract
Hemophilia patients consume large quantities of biologicals. Many are on prophylactic regimens which cost substantially more and have better outcomes as compared to on-demand treatment. In either situation, 95% of costs of care are biologics. As costs are high, third-party payers frequently challenge various treatment regimens. Aetna Inc., working with nurses and a hemophilia treater, initiated a systematic evaluation program of 400 members with hemophilia enrolled in HMO plans to investigate whether these costs could be appropriately controlled. Several caveats were agreed to:
No therapeutic modality would be changed unless medically indicated. Patients would be supported to have an annual visit to a comprehensive center, if they were not already followed by one. Changes where biologics were purchased were only done with physician and client agreement.
Over the past three years, substantial savings have been accomplished PMPM (per member / per month fully-insured HMO member).
The annual spending PMPM for biologics PMPM Total dollars Before program 2003 $0.66 25,650,021 Year 1 2004 $0.60 23,832,539 Year 2 2005 $0.54 21,309,444 Year 3 2006 $0.54 19,395,661
The savings were attributable, in the main, to simply shifting how and where biologics were purchased and could not have been accomplished without the federal PHS pricing program. In three years’ time, annual savings per patient were substantial. The savings per subscriber encouraged Aetna, Inc. to continue this program.
Glanzmann Thromboasthenia with Isolated Amegakaryocytic Thrombocytopenia.
Authors: Corinaldesi Giorgio , Corinaldesi Christian 
Published: April 2020

Abstract
Glanzmann thromboasthenia is an autosomal recessive bleeding disorder caused by a quantitative and/or qualitative defect of the platelet glycoprotein GPalfaIIb/betaIIIa (IIb/IIIa integrin ITGA2B-ITGB3) complex. We have studied a 24 years old patient (exon 2113 T-C substitution missense C705F/C674R) with refractory bleeding platelet count above 25.000, and a very prolonged bleeding time, extensive mucocutaneous petechiae, nosebleeds, and bleeding gums, marked spenomegaly, and conjunctival hemorrage with a prevalence of HLA-DRw2 and DRB1-0410 and a severely decreased platelet production. Immunophenotyping of lymphocytes showed a relative increase of the subset of gamma/delta T-cell receptor (TCR) positive, and CD4 CD8 negative T lymphocytes; expression of c-Mpl and TPO were down-regulated. The bone marrow examination revealed extremely scarce and small megakaryocytes (with a great reduction of CD34+ and CD41+ and a small fraction of CD42+) and with normal marrow cellularity regarding other cellular lines; chromosomal analysis was also normal, while there was an associated severe anaemia with elevated HbF without any history of alcohol use or drugs ingestion. The important observation in this case was the need of many platelet trasfusions, and of fresh frozen plasma (FFP); the functional consequences of this kind of therapy include the induction of abnormalities of the hormones or their receptors (TPO/c-Mpl) that can be decreased in these patients; in addition, platelet mass blocks the biologic effects of SDF-1/CXCR4 receptor and the degree of this inhibition is correlated inversely with platelet count, and supports the evidence that the thrombocytopenia may be related to the inhibition of transcription factors (GATA-1, NF-E2, TAL-1, ETS family), and it directly correlates with the excess of platelet transfusions as an inducible knock-out in MK differentiation. Moreover, this findings indicate that when the receptors (c-Mpl) is saturated, the JAK-2 activation is neutralized and subsequent phosphorylation of STAT-3 and STAT-5 are down-regulated. This patient responded to treatment with danazol 200 mg daily and desmopressin (DDAVP) 0.3 mg/kg, while an additional therapy with traxenamic acid and recombinant FVIIa reduced mucosal bleeding.
The Effectiveness of Recombinant Factor VIIa in the Treatment of Bleeding in Pediatric Patients: A Ten-Year Experience from 1997 to 2007.
Authors: Chuansumrit Ampaiwan , Teeraratkul Sumate , Tripongkaruna Suporn , Wanichkul Suthep , Sirachainan Nongnuch , Pakakasama Samart , Nuntnarumit Pracha , Hongeng Suradej 
Published: April 2020

Abstract
Introduction: Recombinant factor VIIa (rFVIIa) has been used in the treatment of bleeding in Thailand since 1997.
Methods: 125 patients receiving rFVIIa at the Faculty of Medicine, Ramathibodi Hospital and Regional Hospitals from 1997 to 2007 were retrospectively evaluated. Their age group ranged from newborn (n=11), infants (n=21), children (n=86) to adolescents (n=7). They were divided into 2 groups. Group 1 included 11 hemophiliacs with high inhibitor (A10, B1) exhibiting 90 episodes of bleeding or undergoing surgeries. The rFVIIa 100–200 μg/kg at 1–2 h interval for 1–4 doses combined with prednisolone, transamine, fibrin glue and dental splint were given. In cases of surgery, 100 μg/kg at 1–2 h interval was given in the first 24 h and extended to 3–4 h until the removal of stitches or 7–14 days after the surgical procedures. Sequential use of rFVIIa with non-activated or activated prothrombin complex concentrates were given to 9 patients. Group 2 included 114 non-hemophiliacs with or without pre-existing hemostatic disorder for controlling excessive bleeding unresponsive to conventional blood components or for preventing bleeding in invasive procedures. There were 122 episodes involving patients with Dengue Hemorrhagic Fever (n=38), massive bleeding (n=49), invasive procedure (n=23), Glanzmann thrombasthenia (n=6), congenital factor VII deficiency (n=5) and acquired aplastic anemia (n=1). For controlling excessive bleeding, 40–100 μg/kg of rFVIIa was given at 15–30 min interval for 1–4 doses until the bleeding significantly reduced, followed by 40–100 μg/kg at 2–4 h interval for 24–48 h until the bleeding was completely ceased. For preventing bleeding in invasive procedures, the similar regimen but with a lower fixed dose of 40–100 μg/kg was used. One to two doses were usually used. An exceptional high dose of 200–300 μg/kg was used in patients with Glanzmann thrombasthenia and a lower dose of 20–30 μg/kg at 6 h interval was used in patients with congenital factor VII deficiency.
Results: The efficacy of controlling bleeding in hemophiliacs revealed effective (80/90, 88.9%), partially-effective (6/90, 6.7%) and ineffective (4/90, 4.4%) responses while the non-hemophiliacs revealed effective (81/122, 88.5%), partially-effective (14/122, 11.5%) and ineffective (27/122, 22.1%) responses. The case-fatality rates were 0% and 27.2% (31/114) in hemophiliacs and non-hemophiliacs, respectively. Three episodes of thrombosis (3/125, 2.4%) were found 3 children underwent cardiac surgery.
Conclusion: rFVIIa has shown an effectiveness in the treatment of bleeding in pediatric patients with hemophiliacs and non-hemophiliacs.
Management of High Haemorragic Risk Surgery: A Case of Acquired von Willebrand Disease (AVWD).
Authors: Mostarda Giovanni , Cafro Anna Maria , Caimi Teresa Maria , Redaelli Rosaria , Corno Anna Rosa , Sambruni Ivano , Morra Enrica 
Published: April 2020

Abstract
Introduction: Acquired von Willebrand Disease (AVWD) is a rare bleeding disorder occurring in patients with haematological malignancies, solid tumours, immunological and cardiovascular disorders and monoclonal gammopathy of undetermined significance (MGUS). Several pathogenetic mechanisms have been proposed to explain acquired von Willebrand factor (VWF) defect including autoantibodies. Muco-cutaneous bleeding and laboratory findings are similar to those of the congenital VWD. Several approaches have been attempted to correct haemostatic abnormalities with the aim to control bleeding diathesis or to prevent bleeding during surgery. Desmopressin, plasma-derived FVIII/VWF concentrates, intravenous high dose immunoglobulin (HDIg) are the most commonly used. The efficacy of therapy is not predictable and has to be tested before surgery or invasive procedures. In several cases more than one therapeutic option is required. Case report. The patient was a 64-years-old woman with IgGλ-MGUS. Because of persistent uterine bleeding a neoplasia was suspected. The pre-operative coagulative screening showed a prolonged aPTT with markedly reduced levels of FVIII and VWF-related activities (Table 1). VWF multimeric analysis evidenced loss of high and intermediate molecular weight multimers. Patient’s personal and familial history was negative for bleeding and VWF level was normal in other family members. Desmopressin administration (s.c.0.3 μg/kg) obtained a rapid but transient correction of plasma VWF-related parameters, with a return to baseline levels within 6 hours. HDIg intravenous infusion (1 g/kg/day for two consecutive days) led to stable normalization of VWF:RCo, VWF:Ag and FVIII for at least 7 days (Table 1). A laparoscopic hystero-annessiectomy was performed after two-day HDIg infusion: no bleeding complications were observed in the peri-operative period. Histological diagnosis was endometrial hyperplasia. During the two-months follow-up no haemorrhagic episodes were reported.
Comments: In AVWD the correction of laboratory abnormalities is crucial to prevent bleeding in patient undergoing high haemorrhagic risk surgery. The choice among therapeutic options is empirical and depends on underlying disease and clinical setting. The efficacy has to be verified. In our patient HDIg infusion obtained a sustained normalisation of VWF-related parameters and prevented bleeding during surgery and in the post-operative period.
Table 1. Patient’s laboratory parameters before and after treatment with HDIg VWF-related parameters Reference interval basal +1* +5* +7* *days after HDIg infusion aPTT (R) 1.73 1.21 1.21 1.27 &lt;1.19 VWF:RCo (U/dL) &lt;8 59 56 50 46-118 VWF:Ag (U/dL) 11 66 91 85 43-145 FVIII (U/dL) 14 96 85 96 51-164
von Willebrand Disease in Chronic Immune Thrombocytopenic Purpura Children.
Authors: Lee Ji Hye , Yoon Hwi-Joong , Lee Kun Soo 
Published: April 2020

Abstract
Purpose: von Willebrand disease (vWD) is the most common hereditary bleeding disorder and type 2B combines thrombocytopenia. So it must be considered in patients found to have low platelet counts, particularly if there is a family history of mucocutaneous hemorrhage. We performed this study to diagnose the type 2B vWD in chronic immune thrombocytopenic purpura (ITP) children.
Methods: Seventeen cases among chronic ITP children over 6 months at the Department of Pediatrics, Kyungpook National University Hospital, Daegu, Korea from October, 1995 to June, 2007 were participated in this study. We performed screening coagulation tests such as platelet counts, activated partial thromboplastin time (aPTT), and bleeding time (BT) and specific tests for vWD such as von Willebrand factor related antigen (vWF: Ag), vWF ristocetin cofactor (vWF: RCo), factor VIII, and vWF multimer on these patients. These tests were also performed their family when the patient was diagnosed vWD. And we reviewed their past and family histories about bleeding tendency.
Results: There were four boys and thirteen girls and their mean age was 11.6 years (range: 2.8∼18.5 years). Five cases (5/17, 29.4%) were diagnosed vWD: one had lower level of vWF: RCo and factor VIII with normal level of vWF: Ag and others had lower level of vWF: RCo and vWF: Ag with normal level of factor VIII. Among these, two cases showed abnormal screening test results, prolongation of aPTT or BT. We could perform the vWF multimer test in two cases, but two had normal pattern. Among five vWD children, we could obtain the past and family bleeding tendency histories except one case and three families showed bleeding tendency. But all families showed normal screening and specific test results.
Conclusions: von Willebrand disease was combined in 5 cases (29.4%) among 17 chronic thrombocytopenic children. More evaluation such as vWF multimer and ristocetin induced platelet aggregation test (RIPA) is needed to confirm the subtype. And we should repeat the evaluation to the family who had bleeding history but showed normal results for diagnosis or exclusion of vWD.
Redistribution of Recombinant Activated Factor VIIA into Platelets and Vascular Bed: Implications on Bioavailability and Hemostatic Mechanisms.
Authors: Galan Ana Maria , Lopez-Vilchez Irene , Gracia Juan , Escolar Gines 
Published: April 2020

Abstract
Background: Clinical evidence suggests that hemostatic action of recombinant activated factor VII (rFVIIa) exceeds its predicted plasma half life (around 3 hours). Mechanisms involved in the long lasting effects of rFVIIa for prophylactic treatment of patients with hemophilia and inhibitors have not been fully elucidated. Previous studies from our group have demonstrated that platelets internalize tissue factor preparations containing small amounts of FVII. We have investigated the possible redistribution of rFVIIa in different intracellular compartments focussing more specifically on platelets, endothelium and subendothelial matrix.
Methods: We exposed platelet rich plasma, isolated platelets, human umbilical veins and endothelial cells in culture (HUVECs) to rFVIIa. Samples were incubated for up to 2 hours at rFVIIa concentrations from 2-60 mg/ml (normal plasma concentration: 0.5 mg/ml). Flow cytometry techniques were applied to detect surface and intracellular antigens, including FVIIa, in platelets before and after exposure to membrane permeabilizing agents. Immunocytochemical techniques were applied to detect FVIIa in cross sections of umbilical veins and confocal microscopy was used to detect FVIIa in HUVECs. In additional studies, aliquots of washed platelets previously exposed to rFVIIa were incorporated into whole blood anticoagulated with low molecular weight heparin (LMWH) and perfused through a collagen rich damaged vascular surface at a shear rate equivalent to 600/sec.
Results: Flow cytometry studies revealed a significantly enhanced presence of intracellular FVIIa in platelets previously exposed to rFVIIa. Fluorescence intensity related to FVIIa was dependent on the concentration of rFVIIa used during the incubation. A more intense labeling for FVIIa was observed in the subendothelial matrix of umbilical veins exposed to rFVIIa. In perfusion studies, presence of platelets previously exposed to rFVIIa improved platelet thrombus formation and enhanced fibrin generation with respect to parallel experiments using platelets not exposed to rFVIIa.
Conclusion: Our results indicate that rFVIIa can be internalized into platelets and redistributed into subendothelial compartments. Redistribution of rFVIIa into other cellular compartments may explain the prolonged prophylactic action of rFVIIa in some clinical conditions. The existence of extravascular sources of FVIIa may provide new insights into the physiological and pathological implications of FVIIa on hemostasis mechanisms.