bluuebunny/crossref_metadata_2025_split huggingface dataset:
Xanthine Oxidase Mediates High Fat Diet-Induced Vascular Permeability via Junction Adhesion Molecule a Tyrosine Phosphorylation and Tight Junction Disruption
Authors: Chattopadhyay Rima , Kundamani-Sridharan Venkatesh , Tinnikov Alexander , Dyukova Elena , Singh Nikhlesh K , Kotla Sivareddy , Rao Gadiparthi N 
Published: October 2019

Abstract
Previously we have shown that 15(S)-hydroxyeicosatetraenoic acid (15(S)-HETE), the 15-LO metabolite of arachidonic acid (AA), affects endothelial cell (EC) barrier function. In this study, we have examined the role of xanthine oxidase (XO) in 15(S)-HETE-induced EC barrier dysfunction. 15(S)-HETE induced JamA phosphorylation on tyrosine residues 164, 218 and 280 involving XO-mediated reactive oxygen species (ROS) production and Src and Pyk2 activation, resulting in its dissociation from Occludin and causing tight junction disruption with increased EC barrier permeability and enhanced leukocyte and monocyte transmigration in vitro using EC monolayer and ex vivo using arteries as models. The phosphorylation of JamA on tyrosine residues 164, 218, and 280 appear to be critical for its role in 15(S)-HETE-induced EC barrier dysfunction, as mutation of any one of these amino acid residues prevented its dissociation from Occludin and restored TJ integrity and barrier function. Feeding WT mice but not 12/15-LO-/- mice with HFD for 3 months resulted in enhanced XO expression and its activity in the artery, which was correlated with increased aortic EC TJ disruption and its barrier permeability with enhanced leukocyte adhesion. These observations provide additional support for the role of 12/15-LOX in the dysregulation of vascular permeability in response to HFD.

Disclosures
No relevant conflicts of interest to declare.
Subcutaneous Immunoglobulin (SCIG) in Responders to Intravenous Therapy with Chronic Immune Thrombocytopenia (ITP)
Authors: Iuliano Francesco , Iuliano Eleonora , Luci Maria , Perricelli Alessia , Pomillo Angelo , Abruzzese Elisabetta 
Published: October 2019

Abstract
Introduction. Immune thrombocytopenia (ITP) is characterized by platelet destruction due to the presence of platelet antibodies. This phenomenon is associated with impaired platelet production by bone marrow. Short-course corticosteroids and high-dose intravenous immunoglobulins remain the first-line treatments. Aim of this study was to test if SCIG is equally effective and can be less expensive than hospital-based IVIG in treating chronic-ITP
Methods
Seven-ITP patients aged between 13-52 years were given liquid intravenously immunoglobulin (IVIg) stabilized with proline in a dose of 400 mg/kg per day for 5 consecutive days.The median time since ITP diagnosis was 18 months (range, 16 – 38) Median (range) baseline platelet count 19 (3 – 32) x 109/L.,median of 2 (1 – 3) prior ITP therapies. Furthermore, none of of the subjects was a good candidate for TPO-receptor agonists (romiplostim and eltrombopag) because of age or other comorbidities.6 / 6 had a good response to the first 5 day course of gammaglobulin therapy, (platelet count ≥50 × 10(9) /l and ≥2× baseline). The peak platelet count occurred within 7 days from the beginning of the therapy. Only in one patient the platelet count peak took place after 12 days. During the first year of treatment, only one of seven initial responder patients achieved CR(defined as platelet count ≥ 100 * 10^9/L measured on two occasions > 7 days apart and the absence of bleeding) and was excluded from the study. After a loading dose of 2 g/kg over 2 days of IVIg, home SCIG of 8 gr total weekly dose was started by 87,3% (6/7) to maintain platelet counts in the target range of 50–100x109/L.
Results
5/6 patients reached the target. One patient achieved CR during the maintenance therapy.No untoward reactions necessitating cessation of therapy were encountered during this study. The most common side effect observed was headache.
Conclusion
Application of SCIG was well tolerated, easy to manage, and led to stabilization of the disease course.The therapy facilitates home therapy, as the infusion technique is easy for children, adults and elderly people to learn and there is no need for venous access. SCIG home therapy leads to significantly improved life situations for the patients; the SCIG home therapy regimen in particular reduces the costs of treatment. Overall costs per patient were strongly reduced in SCIG in comparison to IVIg.SCIG may represent an effective new therapeutic option in chronic ITP.

Disclosures
Abruzzese: novartis, bristol myers squibb, ariad, pfizer, takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees.
five Int22h Homologous Copies in Association with Intron22 Inversion Type 3
Authors: Lannoy Nathalie , Ravoet Marie , Grisart Bernard , Fretigny Mathilde , Hermans Cedric R. 
Published: October 2019

Abstract
Introduction
F8 intron22 inversion is the causative gene defect in up to 45% of severe hemophilia A (HA) patients mediated by recombination between three highly homologous copies located in intron 22 (int22h-1) and two other extragenic copies (int22h-2 and int22h-3) positioned more telomerically outside the gene. Intrachromosomal rearrangement between int22h-1 and int22h-3 provide the F8 type 1 inversion while the F8 type 2 inversion could be explained by the participation of the int22-h2 sequence in the homologous recombination. However, a third type, known since 1993*, was explained by the existence of a duplicated copy of int22h-3 or int22h-2 in case of intron22 inversion type 3A or 3B respectively.
Methods
Three unrelated HA patients with intron22 type 3A/3B were identified by southern blotting. To appreciate the length of the int22h extragenic duplicated, genomic hybridisation was performed using Affimetrix CytoScan High-Density array.
Results
Breakpoint analyses by CGH-array in all patients show same duplication of approximately 180kb delimited between intron 1 of CLIC2 gene and distal repeat int22h-3reflecting the presence of five genomic int22 copies. A fourth non-hemophiliac case was added since analysis by CGH technique identified duplication delineated by same boundaries.
Conclusion
This study suggests the existence of genotypes harboring five int22h copies mediated by 180 kb duplication at Xq28 locus probably not associated with HA. We propose that this duplication has happened by tandem inversed duplication. Such genotype is to be considered as polymorphism and could be associated with the two kinds of F8 intron22 inversion type 3 when intrachromosomal recombination has occurred between homologous copies.
*Antonarakis SE and the international consortium study. Factor VIII gene inversions in severe hemophilia A: results of an international consortium study. Blood 1995; 86: 2206-2212

Disclosures
No relevant conflicts of interest to declare.
Influence of Factor V Deficiency on ROTEM® Clotting Time
Authors: Haas Thorsten , Cushing Melissa M. , Asmis Lars M 
Published: October 2019

Abstract
Introduction
Acquired factor V (FV) deficiency is repeatedly mentioned to be a major driver of trauma induced coagulopathy. Viscoelastic point-of-care tests such as ROTEM® seem to be advantageous in detection and guidance of trauma related bleeding management. Physiologic effect of FV can be characterized by an increase in thrombin building, which can be detected by ROTEM® clotting time (CT). However, the impact of varying levels of factor V activity in otherwise normal plasma on ROTEM® CT is not yet published. The aim of this in vitro investigation is to assess if mild to moderate factor V deficiency can be detected by ROTEM® CT.
Methods
Factor V deficient plasma (Code 0020011500; Instrumentation Laboratories, Bedford, MA, USA) and normal human plasma (pooled, FV content 96%) was mixed to generate plasma with 0, 10, 16, 25, 50 and 100% FV content. The FV deficient plasma is rendered FV deficient by immunologic means and has FV activity of ≤1%.
ROTEM® (TEM International GmbH, Munich, Germany) measurements were performed in each plasma dilution to assess impact of different levels of FV on thrombin generation. Measurements were repeated in triplicate. Clotting time (CT) was plotted against FV activity. Data is depicted as mean +/- SD.
Results
ROTEM® measurements showed a prolongation of clotting time with decreasing FV activity. A decrease of factor V activity from 100% to approximately 25% was accompanied by a slight change in CT from 50 sec to approximately 65 sec. If factor V activity drops significantly to less than 10%, a much more pronounced prolongation in CT of more 140 sec was observed.
Discussion
Most publications dealing with acquired factor V deficiency following trauma related bleeding reference a prospective study demonstrating that factor V becomes critically low in about 20% of all cases. Those levels of below 30% factor activity were linked to an increase in TEG® r time, which is comparable to the ROTEM® CT, similar to the results in our study.
Notably, a case control study demonstrated that factor V remained fairly stable (77% ± 29%) in trauma patients with no decrease in thrombin generation. Likewise, another case control study in trauma patients demonstrated that EXTEM CT remained within reference range (median 64 sec (IQR 55,75 sec)).
Conclusion
Moderate to severe factor V deficiency can be detected by prolongation of ROTEM® CT in platelet poor plasma (PPP). Further studies will help to determine if platelets added to PPP may further decrease ROTEM® CT, as approximately 20% of the entire FV pool is stored in platelet α-granules in a partially activated form. In addition, the occurrence and the potential clinical impact of factor V deficiency in trauma related bleeding management needs to be further investigated.

Disclosures
Haas: CSL Behring: Honoraria; Octapharma: Honoraria; TEM International: Honoraria.
Delivering a Hemophilic Newborn with No Intracranial Hemorrhage: Is Caesarian Section the Answer?
Authors: Nazir Hanan , Allawati Tuqa , Elbeshlawi Ismail , Al-Khabori Murtadha K. , Alkindi Sara , Alriyami Wafa , Wali Prof. Yasser 
Published: October 2019

Abstract
Birth is not only considered the first hemostatic challenge for a child with hemophilia A, but also it can result in one of the most devastating and often preventable type of bleeding in hemophilic newborns. Three to four percent of infants with hemophilia experience a head bleed at delivery. The optimal mode of delivery of the hemophilic carrier expecting an affected child is still a matter of uncertainty and debate.
The aim of this study was to evaluate the frequency of major bleeding in the neonatal period and to examine the association between mode of delivery and neonatal intracranial hemorrhage (ICH) in an Arab cohort of infants with hemophilia A.
Subjects and methods:
A retrospective multi-center study of children with severe hemophilia A (age 0-18 years) was conducted. Five hemophilia treatment centers distributed across four Arabian Gulf countries participated in the study. Two centers from Oman and one center of each of KSA, UAE and Kuwait were included.
The collected data were date of birth, maternal parity, mode of delivery, duration of labor, admission to neonatal intensive care unit, symptomatic ICH, and other bleeding episodes. Review of electronic patient record along with telephone interviews when needed were the tools used for data collection.
Results:
A total of 153 patients were found. Most of them were born through spontaneous vaginal delivery (SVD) (125 patients; 82 %). Vacuum extraction was adopted in six patients (4%), while twenty two patients were offered caesarian section (CS) (14%).
ICH was reported in a total of four patients (2.6%). All of them were full term babies with variable time interval between the onset of the bleed and birth. One patient presented at birth, another presented at the age of 4 weeks while the remaining two had the bleed at the age of 2 weeks.
Vacuum extraction was associated with the highest risk of ICH (two patients; 33 %).The other two patients were born by SVD (1.6 %). CS was not associated with ICH.
Conclusions:
Normal vaginal delivery is still considered a safe journey through the birth canal for hemophilic newborns. Larger prospective studies are needed to define an evidence-based optimal mode of delivery for the hemophilia carrier expecting an affected child.

Disclosures
No relevant conflicts of interest to declare.
Evaluation of Thrombin Generation Assay in Patients with Hemophilia
Authors: Karimi Mehran , Haghpanah Sezaneh , Bazrafshan Asghar , Eatemadfar Peyman , Dehghani Javad , Afrasiabi Abdolreza 
Published: October 2019

Abstract
Background: Diagnosis of hemophilia is based on detection of factor level and coagulation screening test. These screening tests can not reflect global hemostatic balance. Thrombin generation (TG), as a focal point and key process, is an important step in hemostasis and thrombosis. Recent evidence has indicated that TG assay is a better predictor of the coagulation capacity and subsequently overall assessment of hemostasis compared with traditional coagulation tests. The parameters of the thrombogram are helpful in the evaluation of bleeding or thrombotic risk and also in the management of these situations.
Objective: The aim of this study is to evaluate the correlation between TG parameters with bleeding symptoms and disease severity in patients with hemophilia.
Methods: In this cross-sectional study, 59 male patients with hemophilia were randomly selected from those already registered at Shiraz Hemophilia Center of southern Iran, from February to December 2012. Also, 38 healthy age-matched men were considered as a control group from those referred for check up. Informed written consent was obtained from all participants. The proposal was approved by the Medical Ethics Committee of Shiraz University of Medical Sciences. Disease severity was defined based on the activity level of deficient factor (severe <1%, moderate from 1 to 5%, and mild >5% IU/dL). Bleeding score (BS) was calculated by performing a clinical evaluation using a modified questionnaire based on Tosetto et al- questionnaire. Correlation of TG parameters with disease severity and BS were determined.
Results: From 59 patients with hemophilia (52 hemophilia A and 7 hemophilia B), 40 patients (68%) had severe disease, 14 (24%) moderate and 5 (8%) mild disease. All TG parameters showed statistically significant differences between patients and controls (P<0.001). Only Lag time showed a statistically significant correlation with BS (rs= 0.316, P =0.016). All TG parameters except peak showed association with disease severity (P<0.05). In addition, endogenous thrombin potential showed a significant correlation with factor activity level (rs= 0.459, P <0.001). Both Lag time and start tail showed significant negative correlations with factor activity level (rs= -0.488, P <0.001 and rs=- 0.289, P <0.026 respectively).
Conclusion: Based on our results, although most of the TG parameters evaluated were not significantly correlated with bleeding score of hemophilia patients, the majority of TG parameters were significantly associated with factor activity level and disease severity. In patients with hemophilia, plasma factor activity level has a poor predictive value in the evaluation of efficacy of treatment; but it seems that TG assay is an appropriate tool for assessment of global hemostasis and better reflection of clotting function in the management of patients with hemophilia.

Disclosures
No relevant conflicts of interest to declare.
Severe Spontaneous Intracranial Bleeding in 11 Months Old Boy with Congenital Afibrinogenemia with Novel Mutation - Deletion aα 6477A in Fibrinogen Gene
Authors: Zapletal Ondrej , Blatny Jan , Kotlin Roman , Dyr Jan E 
Published: October 2019

Abstract
Congenital afibrinogenemia is very rare inherited bleeding disorder, caused by absence of fibrinogen in plasma. Serious spontaneous bleeding including intracranial haemorrhage may occur at any age.
We present a case report of a boy with afibrinogenemia caused by novel mutation – homozygous deletion Aα 6477A. Diagnosis was set after bleeding post cleft lip and palate plastic surgery. Spontaneous intracranial haematoma in occipital region occurred when the boy was 11 month old. Replacement therapy with plasma derived fibrinogen concentrate successfully covered neurosurgical evacuation of hematoma. The patient was then commenced on regular prophylaxis with fibrinogen concentrate two times a week. No further bleeding episodes occurred until present time.
Spontaneous intracranial bleeding in patients with inherited bleeding disorders is often life threatening. Immediate replacement therapy is crucial, together with diagnostics and surgery, when necessary. Prophylactic substitution therapy after a severe bleeding episode is recommended.

Disclosures
No relevant conflicts of interest to declare.
Sub-Clinical Activation of Thrombosis May Predict Survival in Cancer Patients
Authors: Batra Anupam , Mehdi Syed A. , Brodzik Frank , Pasquale Donald 
Published: October 2019

Abstract
Background: Approximately 10% of newly diagnosed cancer patients will develop thromboembolic disease (TED) in the first year following diagnosis. We hypothesize that a subgroup of cancer patients at diagnosis will have subclinical changes in their coagulation system that will indicate increased risk for subsequent TED and/or predict increased adverse cancer outcome.
Methods: We performed a prospective, non-interventional study of recently diagnosed cancer patients who were scheduled to start chemotherapy. We obtained a single plasma sample at the time of diagnosis and prior to therapy, and measured prothrombin 1.2 fragment (F1.2), a measure of thrombin activation, and plasma microparticle levels (MP), a measure of platelet, endothelial, and other cell activation. We reviewed subsequent clinical outcomes including survival and incidence of venous thrombosis. We accrued 22 patients with mean age 63 years (range 39 to 89; 20 males, 2 females), 12 with lung, 6 with colorectal, and 4 with head and neck cancer. Mean follow-up was 24 months (range 3 to 57). Plasma F1.2 and MP were assayed using commercially available ELISA kits.
Results:
T1.2 concentrations were significantly higher in the deceased subgroup (table 1; p<0.002), and in patients with venous thrombosis (table 2; p<0.05). MPs were not informative. Sensitivity and specificity for prediction of survival and venous thrombosis (table 3) were determined by comparing patients in highest quartile of levels with lower quartiles combined and by using threshold value of mean +2.5 SD’s determined by measuring levels in 8 healthy controls.
Table 1. Survival: F1.2 and MP (Mann-Whitney) Deceased Patients (N=7) Alive Patients (N=15) Mean +/- SD Median Mean +/- SD Median p-value F1.2 (pM) 436+/-214 355 121+/-117 105 <0.002 MP (nM) 10+/-10 3 8.8+/-2.8 2.8 Not Significant
Table 2. Venous Thrombosis: F1.2 and MP (Mann-Whitney) Thrombosis (N=4) No Thrombosis (N=18) Mean +/- SD Median Mean +/- SD Median p-value F1.2 (pM) 368+/-267 313 189+/-191 163 <0.05 MP (nM) 5.9+/-5.3 3.9 9.8+/-15.6 2.7 Not Significant
Table 3. Sensitivity/Specificity Analysis >mean + 2.5 SD’s Highest Quartile MP T1.2 MP T1.2 Death SENSITIVITY 43% 43% 43% 71% Death SPECIFICITY 87% 100% 87% 93% Thrombosis SENSITIVITY 25% 25% 25% 50% Thrombosis SPECIFICITY 78% 89% 78% 78%
Conclusion: T1.2 levels measured at time of diagnosis of cancer correlated with increased mortality and development of venous thrombosis. MP levels were not informative, possibly due to the limited size of the study population. Additional studies are needed to elucidate whether these biomarkers may be useful in identifying a higher risk population for death and/or venous thrombosis, and to help guide enhanced therapy for such higher risk patients.

Disclosures
No relevant conflicts of interest to declare.
Flow Cytometry Screening for Paroxysmal Nocturnal Hemoglobinuria Abnormal Clones in Saudi Arabia
Authors: AlGhasham Nahlah , Abulkhair Yasmeen , Khalil Salem 
Published: October 2019

Abstract
Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disease with insidious process, chronic course and life-threatening condition. PNH is clinically defined by the deficiency of the endogenous glycosyl phosphatidylinositol (GPI)-anchored complement inhibitory protein. It has always aroused interest in the medical profession rendering screening and proper diagnosis by flow cytometry (FCM) technology a priority
We reviewed all samples submitted for PNH/ FCM screening for the past 2 years (2012-2013) at hematology section, Department of Pathology and Laboratory Medicine, King Faisal Specialist Hospital and Research Center (General Organization). We collected the positive cases and reviewed them for age, gender, indication for screening, sample type, size of the PNH clone and cell type affected. Immunophenotypic analysis was performed using gating antibodies CD45, CD15, CD33, CD235a GPI-linked antibodies, CD59, CD14, and CD24 as well as fluorescent Aerolysin (FLAER).
In a total of 366 peripheral blood samples submitted for PNH/ FCM screening fifteen samples (4%) were positive for PNH clones but only 12 patients were available for analysis. The median age for our patients was 34 years with approximately equal male to female distribution.
12 cases showed type II and III clones within the RBCs with clone size ranging between 0.04% and 56%. Analysis of granulocytes and monocytes revealed type III clone in 8 cases, type II and III clone in 3 cases and non in one case. The percentage of the clone varies between the granulocytes and monocytes and ranges from 1% up to 100%.
Of 12 positive PNH cases, 8 (66.7%) patients were diagnosed as having aplastic anemia (AA), 1 (8.3%) patient with Budd-Chiari syndrome, 1 (8.3%) patient has chronic immune thrombocytopenia (ITP), and 2 (16.7%) patients presented with pancytopenia.
This study confirms the rarity of the disease since only 4% of the submitted samples for analysis turned to be positive for PNH. The detection limit for a PNH clone by FCM in the RBC or WBC is 0.01%. Identification of small PNH clone is greater FCM sensitivity relative to old test used for the same purpose (Ham test). The use of the FLAER allowed us to detect granulocytic PNH clone, however, granulocytes PNH clone detection alone without RBCs clone detection is not recommended. This review confirms the previous percentage of positive cases (5.9%) reported from this center on a smaller number of cases during the past few years.

Disclosures
No relevant conflicts of interest to declare.