bluuebunny/crossref_metadata_2025 huggingface dataset:
O-GlcNAcylation of FoxO1 mediates nucleoside diphosphate kinase B deficiency induced endothelial damage
Authors: Shan Shenliang , Chatterjee Anupriya , Qiu Yi , Hammes Hans-Peter , Wieland Thomas , Feng Yuxi
Published: July 2018

Nucleoside diphosphate kinase B (NDPK-B) acts as a protective factor in the retinal vasculature. NDPK-B deficiency leads to retinal vasoregression mimicking diabetic retinopathy (DR). Angiopoetin 2 (Ang-2), an initiator of retinal vasoregression in DR, is upregulated in NDPK-B deficient retinas and in NDPK-B depleted endothelial cells (ECs) in vitro. We therefore investigated the importance of Ang-2 in NDPK-B deficient retinas and characterized the mechanisms of Ang-2 upregulation upon NDPK-B depletion in cultured ECs. The crucial role of retinal Ang-2 in the initiation of vasoregression was verified by crossing NDPK-B deficient with Ang-2 haplodeficient mice. On the molecular level, FoxO1, a transcription factor regulating Ang-2, was upregulated in NDPK-B depleted ECs. Knockdown of FoxO1 abolished the elevation of Ang-2 induced by NDPK-B depletion. Furthermore O-GlcNAcylated FoxO1 was found preferentially in the nucleus. An increased O-GlcNAcylation of FoxO1 was revealed upon NDPK-B depletion. In accordance, the inhibition of protein O-GlcNAcylation normalized NDPK-B depletion induced Ang-2 upregulation. In summary, we demonstrated that the upregulation of Ang-2 upon NDPK-B deficiency is driven by O-GlcNAcylation of FoxO1. Our data provide evidence for a central role of protein O-GlcNAcylation in NDPK-B associated vascular damage and point to the hexosamine pathway as an important target in retinal vasoregression.
Voxel size and gray level normalization of CT radiomic features in lung cancer
Authors: Shafiq-ul-Hassan Muhammad , Latifi Kujtim , Zhang Geoffrey , Ullah Ghanim , Gillies Robert , Moros Eduardo
Published: July 2018

Radiomic features are potential imaging biomarkers for therapy response assessment in oncology. However, the robustness of features with respect to imaging parameters is not well established. Previously identified potential imaging biomarkers were found to be intrinsically dependent on voxel size and number of gray levels (GLs) in a recent texture phantom investigation. Here, we validate the voxel size and GL in-phantom normalizations in lung tumors. Eighteen patients with non-small cell lung cancer of varying tumor volumes were analyzed. To compare with patient data, phantom scans were acquired on eight different scanners. Twenty four previously identified features were extracted from lung tumors. The Spearman rank (rs) and interclass correlation coefficient (ICC) were used as metrics. Eight out of 10 features showed high (rs > 0.9) and low (rs < 0.5) correlations with number of voxels before and after normalizations, respectively. Likewise, texture features were unstable (ICC < 0.6) and highly stable (ICC > 0.8) before and after GL normalizations, respectively. We conclude that voxel size and GL normalizations derived from a texture phantom study also apply to lung tumors. This study highlights the importance and utility of investigating the robustness of radiomic features with respect to CT imaging parameters in radiomic phantoms.
Anisotropic transverse magnetoresistance and Fermi surface in TaSb2
Authors: Pariari Arnab , Singha Ratnadwip , Roy Shubhankar , Satpati Biswarup , Mandal Prabhat
Published: July 2018

TaSb2 has been predicted theoretically to be a weak topological insulator. Whereas, the earlier magnetotransport experiment has established it as a topological semimetal. In the previous works, the Shubnikov-de Haas oscillation has been analyzed to probe the Fermi surface, with magnetic field along a particular crystallographic axis only. By employing a sample rotator, we reveal highly anisotropic transverse magnetoresistance by rotating the magnetic field along different crystallographic directions. To probe the anisotropy in the Fermi surface, we have performed magnetization measurements and detected strong de Haas-van Alphen (dHvA) oscillations for the magnetic field applied along a and b axes as well as perpendicular to ab plane of the crystals. Three Fermi pockets have been identified by analyzing the dHvA oscillations. With the application of magnetic field along different crystal directions, the cross-sectional areas of the Fermi pockets have been found significantly different, i.e., the Fermi pockets are highly anisotropic in nature. Three-band fitting of electrical and Hall conductivity reveals two high mobility electron pockets and one low mobility hole pocket. The angular variation of transverse magnetoresistance has been qualitatively explained using the results of dHvA oscillations and three-band analysis.
Thermal conductivity of amorphous SiO2 thin film: A molecular dynamics study
Authors: Zhu Wenhui , Zheng Guang , Cao Sen , He Hu
Published: July 2018

Amorphous SiO2(a-SiO2) thin films are widely used in integrated circuits (ICs) due to their excellent thermal stability and insulation properties. In this paper, the thermal conductivity of a-SiO2thin film was systematically investigated using non-equilibrium molecular dynamics (NEMD) simulations. In addition to the size effect and the temperature effect for thermal conductivity of a-SiO2thin films, the effect of defects induced thermal conductivity tuning was also examined. It was found that the thermal conductivity of a-SiO2thin films is insensitive to the temperature from −55 °C to 150 °C. Nevertheless, in the range of the thickness in this work, the thermal conductivity of the crystalline SiO2(c-SiO2) thin films conforms to the T−αwith the exponent range from −0.12 to −0.37, and the thinner films are less sensitive to temperature. Meanwhile, the thermal conductivity of a-SiO2with thickness beyond 4.26 nm has no significant size effect, which is consistent with the experimental results. Compared with c-SiO2thin film, the thermal conductivity of a-SiO2is less sensitive to defects. Particularly, the effect of spherical void defects on the thermal conductivity of a-SiO2is followed by Coherent Potential model, which is helpful for the design of low-K material based porous a-SiO2thin film in microelectronics.
Generation of multiparametric MRI maps by using Gd-labelled- RBCs reveals phenotypes and stages of murine prostate cancer
Authors: Ferrauto Giuseppe , Di Gregorio Enza , Lanzardo Stefania , Ciolli Laura , Iezzi Manuela , Aime Silvio
Published: July 2018

Prostate Cancer (PCa) is the second most common and fifth cause of cancer-related mortality in males in Western Countries. The development of innovative tools for an early, more precise and noninvasive diagnosis is a medical need. Vascular volume (Vv) and hypoxia are two of the most important tumor hallmarks. Herein, they have been assessed in TRAMP mice by using MRI. Their quantification has been carried out by injecting autologous Red Blood Cells (RBCs), ex vivo labelled with Gd-HPDO3A or Gd-DOTP complexes, respectively. Gd-labelled-RBCs are stably confined in the intravascular space, also in presence of a very leaky tumor endothelium, thus representing efficient probes for vascular space analysis. Vv enhancement and hypoxia onset have been demonstrated to be present at early stages of PCa and their expression largely increases with tumor development. Moreover, also Diffusion weighted MRI and Amide Proton Transfer MRI have been herein applied to characterize PCa. The herein applied multiparametric MRI (mpMRI) analysis allows a detailed in vivo characterization of PCa, in which each histotype and cancer stage displays a specific MRI pattern. This provides an unprecedented opportunity to feature prostate tumor, making possible a non-invasive, precise and early diagnosis, which could direct treatments towards a more personalized medicine.
The energetic behaviour of the human foot across a range of running speeds
Authors: Kelly Luke A. , Cresswell Andrew G. , Farris Dominic J.
Published: July 2018

The human foot contains passive elastic tissues that have spring-like qualities, storing and returning mechanical energy and other tissues that behave as dampers, dissipating energy. Additionally the intrinsic and extrinsic foot muscles have the capacity to act as dampers and motors, dissipating and generating mechanical energy. It remains unknown as to how the contribution of all passive and active tissues combine to produce the overall energetic function of the foot during running. Therefore, the aim of this study was to determine if the foot behaves globally as an active spring-damper during running. Fourteen participants ran on a force-instrumented treadmill at 2.2 ms−1, 3.3 ms−1 and 4.4 ms−1, while foot segment motion was collected simultaneously with kinetic measurements. A unified deformable segment model was applied to quantify the instantaneous power of the foot segment during ground contact and mechanical work was calculated by integrating the foot power data. At all running speeds, the foot absorbed energy from early stance through to mid-stance and subsequently returned/generated a proportion of this energy in late stance. The magnitude of negative work performed increased with running speed, while the magnitude of positive work remained relatively constant across all running speeds. The proportion of energy dissipated relative to that absorbed (foot dissipation-ratio) was always greater than zero and increased with running speed, suggesting that the foot behaves as a viscous spring-damper.
Comprehensive analysis of competitive endogenous RNAs network associated with head and neck squamous cell carcinoma
Authors: Fang Xiao-Nan , Yin Miao , Li Hua , Liang Cheng , Xu Cong , Yang Gui-Wen , Zhang Hua-Xiang
Published: July 2018

Long non-coding RNAs (lncRNAs) can regulate gene expression directly or indirectly through interacting with microRNAs (miRNAs). However, the role of differentially expressed mRNAs, lncRNAs and miRNAs, and especially their related competitive endogenous RNAs (ceRNA) network in head and neck squamous cell carcinoma (HNSCC), is not fully comprehended. In this paper, the lncRNA, miRNA, and mRNA expression profiles of 546 HNSCC patients, including 502 tumor and 44 adjacent non-tumor tissues, from The Cancer Genome Atlas (TCGA) were analyzed. 82 miRNAs, 1197 mRNAs and 1041 lncRNAs were found to be differentially expressed in HNSCC samples (fold change ≥ 2; P < 0.01). Further bioinformatics analysis was performed to construct a lncRNA-miRNA-mRNA ceRNA network of HNSCC, which includes 8 miRNAs, 71 lncRNAs and 16 mRNAs. Through survival analysis based on the expression profiles of RNAs in the ceRNA network, we detected 1 mRNA, 1 miRNA and 13 lncRNA to have a significant impact on the overall survival of HNSCC patients (P < 0.05). Finally, some lncRNAs, which are more important for survival, were also predicted. Our research provides data to further understand the molecular mechanisms implicated in HNSCC.
Outburst floods provide erodability estimates consistent with long-term landscape evolution
Authors: Garcia-Castellanos Daniel , O’Connor Jim E.
Published: July 2018

Most current models for the landscape evolution over geological timescales are based on semi-empirical laws that consider riverbed incision proportional to rock erodability (dependent on lithology) and to the work performed by water flow (stream power). However, the erodability values obtained from these models are entangled with poorly known conditions of past climate and streamflow. Here we use the erosion reported for 82 outburst floods triggered by overtopping lakes as a way to estimate the outlet erodability. This avoids the common assumptions regarding past hydrology because water discharge from overtopping floods is often well constrained from geomorphological evidence along the spillway. This novel methodology yields values of erodability that show a quantitative relation to lithology similar to previous river erosion analyses, expanding the range of hydrological and temporal scales of fluvial incision models and suggesting some consistency between the mathematical formulations of long-term and catastrophic erosional mechanisms. Our results also clarify conditions leading to the runaway erosion responsible for outburst floods triggered by overtopping lakes.
Relationship between Systemic Vascular Characteristics and Retinal Nerve Fiber Layer Loss in Patients with Type 2 Diabetes
Authors: Jeon Soo Ji , Park Hae-Young Lopilly , Lee Jae Hyung , Park Chan Kee
Published: July 2018

Retinal nerve fiber layer (RNFL) loss in diabetic patients is especially common regardless of diabetic retinopathy (DR). The correlations between nonglaucomatous RNFL loss and systemic characteristics in diabetic patients have aroused interests in many aspects. 167 subjects with type 2 diabetes who underwent evaluation for arterial stiffness and cardiovascular autonomic function using heart rate variability (HRV) were included in this study. Arterial stiffness was measured using cardio-ankle vascular index (CAVI) and ankle-brachial index (ABI). Multivariate regression analysis was performed to determine factors influencing the presence of RNFL loss according to age. Factors determining the superior location of diabetic RNFL loss were also investigated. CAVI were worse in patients with RNFL loss, especially in those with old age (≥50 yrs) (p = 0.037). Influential factor of RNFL defect in old group was ABI (p = 0.007). However, in young group (<50 yrs), HRV parameter (low-frequency/high-frequency ratio) determined the presence of RNFL loss (p = 0.040). Significant determinants of superior RNFL defect in old subjects were CAVI and ABI (p = 0.032 and p = 0.024). For young diabetic patients, autonomic dysfunction may have relationship with RNFL loss, but as patients get older, arterial stiffness could aggravate vascular autoregulation and diabetic RNFL loss. RNFL loss in diabetes may be correlated with systemic vascular conditions.
Author Correction: Frequent discussion of insomnia and weight gain with glucocorticoid therapy: an analysis of Twitter posts
Authors: Patel Rikesh , Belousov Maksim , Jani Meghna , Dasgupta Nabarun , Winokur Carly , Nenadic Goran , Dixon William G.
Published: July 2018

The original version of the published Article contained an error in the spelling of the fifth Author’s name. “Carly Winakor” has been changed to “Carly Winokur”. This has been corrected in the HTML and PDF version of the Article.
Direct evidence of the ˙OH scavenging activity of selenium nanoparticles
Authors: Chen Wanwen , Yue Lin , Xia Wenshui
Published: July 2018

Selenium nanoparticle (SeNPs) have been considered as antioxidant agents. The direct evidence for the ˙OH scavenging activity of SeNPs was clearly demonstrated by the chromogenic reaction of MV determined by UV-vis and ESR experiment, which were consistent with the theoretical results.
Sour cherry extract inhibits human salivary α-amylase and growth ofStreptococcus mutans(a pilot clinical study)
Authors: Homoki Judit , Gyémánt Gyöngyi , Balogh Péter , Stündl László , Bíró-Molnár Piroska , Paholcsek Melinda , Váradi Judit , Ferenc Fenyvesi , Kelentey Barna , Nemes Judit , Remenyik Judit
Published: July 2018

Clinical evidence for the effectiveness of a chewing gum with sour cherry extract in inhibiting α-amylase andS. mutans.
Iron promoted C3–H nitration of 2H-indazole: direct access to 3-nitro-2H-indazoles
Authors: Murugan Arumugavel , Gorantla Koteswar Rao , Mallik Bhabani S. , Sharada Duddu S.
Published: July 2018

An efficient C3–H functionalization of indazole has been demonstrated. Notably, this method involves chelation-free radical C–H nitration on 2H-indazole. The radical mechanism was confirmed by control experiments and quantum chemical calculations. The synthetic utility has been proven by the synthesis of bio-relevant benzimidazoindazoles via reductive cyclization.
The self-template synthesis of highly efficient hollow structure Fe/N/C electrocatalysts with Fe–N coordination for the oxygen reduction reaction
Authors: Yu Yue , Xiao Dejian , Ma Jun , Chen Changli , Li Kai , Ma Jie , Liao Yi , Zheng Lirong , Zuo Xia
Published: July 2018

The exploration of highly efficient catalysts to replace noble metal platinum for the oxygen reduction reaction, on which M/N/C catalysts have shed brilliant light, is greatly significant but challenging.
Selective yields of furfural and hydroxymethylfurfural from glucose in tetrahydrofuran over Hβ zeolite
Authors: Tan Jin , Wang Haiyong , Ma Longlong , Wang Chenguang , Liu Qiying , Zhang Qi , He Minghong
Published: July 2018

Several simple and effective solvents combined with Hβ zeolite were tested to selectively convert glucose into furfural and hydroxymethylfurfural in this work.
Auditory Lexical Decisions in Developmental Language Disorder: A Meta-Analysis of Behavioral Studies
Authors: Jones Samuel David , Brandt Silke
Published: July 2018

Purpose

Despite the apparent primacy of syntactic deficits, children with developmental language disorder (DLD) often also evidence lexical impairments. In particular, it has been argued that this population have difficulty forming lexical representations that are detailed enough to support effective spoken word processing. In order to better understand this deficit, a meta-analysis of studies testing children with DLD in the auditory lexical decision task was conducted. The objective was to provide summary effect size estimates for accuracy and response time measures for comparisons to age- and language-matched control groups.

Method

Two thousand three hundred seventy-two records were initially identified through electronic searches and expert consultation, with this cohort reduced to 9 through duplicate removal and the application of eligibility and quality criteria. The final study cohort included 499 children aged 3;8–11;4 (years;months).

Results

Multivariate analysis suggests that children with DLD were significantly less accurate in the auditory lexical decision task than age-matched controls. For the response time estimate, however, confidence intervals for the same group comparison crossed 0, suggesting no reliable difference between groups. Confidence intervals also crossed 0 for language-matched control estimates for both accuracy and response time, suggesting no reliable difference between groups on either measure.

Conclusion

Results broadly support the hypothesis that children with DLD have difficulty in forming detailed lexical representations relative to age- though not language-matched peers. However, further work is required to determine the performance profiles of potential subgroups and the impact of manipulating different lexical characteristics, such as the position and degree of nonword error, phonotactic probability, and semantic network size.
Theories of Working Memory: Differences in Definition, Degree of Modularity, Role of Attention, and Purpose
Authors: Adams Eryn J. , Nguyen Anh T. , Cowan Nelson
Published: July 2018

Purpose

The purpose of this article is to review and discuss theories of working memory with special attention to their relevance to language processing.

Method

We begin with an overview of the concept of working memory itself and review some of the major theories. Then, we show how theories of working memory can be organized according to their stances on 3 major issues that distinguish them: modularity (on a continuum from domain-general to very modular), attention (on a continuum from automatic to completely attention demanding), and purpose (on a continuum from idiographic, or concerned with individual differences, to nomothetic, or concerned with group norms). We examine recent research that has a bearing on these distinctions.

Results

Our review shows important differences between working memory theories that can be described according to positions on the 3 continua just noted.

Conclusion

Once properly understood, working memory theories, methods, and data can serve as quite useful tools for language research.
Gaia Data Release 2
Authors: Holl B. , Audard M. , Nienartowicz K. , Jevardat de Fombelle G. , Marchal O. , Mowlavi N. , Clementini G. , De Ridder J. , Evans D. W. , Guy L. P. , Lanzafame A. C. , Lebzelter T. , Rimoldini L. , Roelens M. , Zucker S. , Distefano E. , Garofalo A. , Lecoeur-Taïbi I. , Lopez M. , Molinaro R. , Muraveva T. , Panahi A. , Regibo S. , Ripepi V. , Sarro L. M. , Aerts C. , Anderson R. I. , Charnas J. , Barblan F. , Blanco-Cuaresma S. , Busso G. , Cuypers J. , De Angeli F. , Glass F. , Grenon M. , Juhász Á. L. , Kochoska A. , Koubsky P. , Lanza A. F. , Leccia S. , Lorenz D. , Marconi M. , Marschalkó G. , Mazeh T. , Messina S. , Mignard F. , Moitinho A. , Molnár L. , Morgenthaler S. , Musella I. , Ordenovic C. , Ordóñez D. , Pagano I. , Palaversa L. , Pawlak M. , Plachy E. , Prša A. , Riello M. , Süveges M. , Szabados L. , Szegedi-Elek E. , Votruba V. , Eyer L.
Published: July 2018

Context. The Gaia Data Release 2 (DR2) contains more than half a million sources that are identified as variable stars.

Aims. We summarise the processing and results of the identification of variable source candidates of RR Lyrae stars, Cepheids, long-period variables (LPVs), rotation modulation (BY Dra-type) stars, δ Scuti and SX Phoenicis stars, and short-timescale variables. In this release we aim to provide useful but not necessarily complete samples of candidates.

Methods. The processed Gaia data consist of the G, GBP, and GRP photometry during the first 22 months of operations as well as positions and parallaxes. Various methods from classical statistics, data mining, and time-series analysis were applied and tailored to the specific properties of Gaia data, as were various visualisation tools to interpret the data.

Results. The DR2 variability release contains 228 904 RR Lyrae stars, 11 438 Cepheids, 151 761 LPVs, 147 535 stars with rotation modulation, 8882 δ Scuti and SX Phoenicis stars, and 3018 short-timescale variables. These results are distributed over a classification and various Specific Object Studies tables in the Gaia archive, along with the three-band time series and associated statistics for the underlying 550 737 unique sources. We estimate that about half of them are newly identified variables. The variability type completeness varies strongly as a function of sky position as a result of the non-uniform sky coverage and intermediate calibration level of these data. The probabilistic and automated nature of this work implies certain completeness and contamination rates that are quantified so that users can anticipate their effects. Thismeans that even well-known variable sources can be missed or misidentified in the published data.

Conclusions. The DR2 variability release only represents a small subset of the processed data. Future releases will include more variable sources and data products; however, DR2 shows the (already) very high quality of the data and great promise for variability studies.
Schulterverletzungen: Risikoscreening und physiotherapeutische Intervention
Authors: Enzler Marcel
Published: July 2018

Schulterverletzungen treten häufig bei Überkopfsportarten wie Handball, Volleyball und Tennis auf. Oft sind diese Verletzungen Folgen chronischer
Fehlbelastungen, die auf Bewegungseinschränkungen und Muskeldysbalancen zurückzuführen sind. Mit einem gezielten Risikoscreening kann man solche Ursachen
identifizieren und mit geeigneten Übungen das Verletzungsrisiko verringern.
Intensivpflege studieren?
Authors: Scheel Yanick-Daniel , Beister Bianca , Mann Marcel , Eggerer Andreas , Wortha-Hoyer Jenny , Pottkämper Karen
Published: July 2018

Ein neuer Studiengang für erweiterte klinische Pflege will seit Ende 2017 Studenten unter anderem in wissenschaftlichem Arbeiten qualifizieren. Doch warum
brauchen wir akademisierte Pflegekräfte am Bett überhaupt? Welche Kompetenzen haben sie nach dem Studium? Und was sagen die Studierenden selbst zu dem neuen
Studiengang? Die Autoren, selbst Studierende des neuen Studiengangs, haben die Studiengangsleitung der Akkon Hochschule, Karen Pottkämper, sowie einige
Kommilitonen dazu befragt. Und Jenny Wortha-Hoyer, stellvertretende Pflegedirektorin der Charité, beschreibt aus Sicht des Krankenhauses ihre Vorstellung vom
Studiengang und schildert, welche Kompetenzen die Absolventen erlangen sollen.
Assessment: Pool Activity Level (PAL) – Aktivitätslevel bestimmen
Authors: Tatzer Verena C. , Pool Jackie
Published: July 2018

Das Pool Activity Level (PAL) Instrument eignet sich für die ergotherapeutische Diagnostik, um die Ressourcen von Menschen mit kognitiver Einschränkung zu erheben. Es unterstützt Therapeuten, Pflegende und Angehörige in verschiedenen Settings, Menschen – zum Beispiel mit Demenz – in bedeutsame Betätigungen zu involvieren.
HeadApp – Hirnleistungstraining 2.0
Authors: Reiter Selina , Striesow Robert
Published: July 2018

Die Software HeadApp eignet sich für das Hirnleistungstraining bei Klienten nach Schlaganfall, Schädel-Hirn-Trauma und anderen Schädigungen des Gehirns. Es kann stationär, ambulant und im Eigentraining zum Einsatz kommen. Selina Reiter und Robert Striesow untersuchten die Anwendung von HeadApp in ihrer Bachelorarbeit und zeigen anhand eines Fallbeispiels aus der Geriatrie, wie sie es in eine betätigungsorientierte ergotherapeutische Intervention integrieren.
Comparison of Three D-Dimer Assays for the Diagnosis of DVT: ELISA, Latex and an Immunofiltration Assay (NycoCard D-Dimer)
Authors: Dale Siri , Gogstad Geir O , Brosstad Frank , Godal Hans C , Holtlund Jostein , Mørk Einar , Brandsnes Øystein , Borch Stig M
Published: July 2018

Ninety-two consecutive patients referred for suspicion of deep venous thrombosis (DVT) were analyzed for D-dimer using ELISA, latex test, and a new immunofiltration method (NycoCard D-Dimer). Contrast venography verified the diagnosis in 40, and excluded the diagnosis in 52 patients. The sensitivity, negative predictive values, specificity and positive predictive values were, for ELISA 98%, 95%, 38% and 54, for NycoCard D-Dimer 100%, 100%, 42% and 57% and for the latex test 73%, 78%, 75%, and 69%, respectively. Sensitivity and specificity were inversely related with increasing pathological cutoff value. Comparison of test results by concentration category revealed a good agreement between ELISA and NycoCard D-Dimer, but to less extent between latex and the two other tests. It is concluded that NycoCard D-Dimer and D-dimer ELISA are well-suited as exclusion tests for DVT. A plasma sample is tested with NycoCard D-Dimer in less than 2 min. Thus, this test combines advantageous analytical properties comparable to the ELISA-test, with rapidity and simplicity comparable to the latex test.
Multicentric Evaluation of a New PT Reagent Based on Recombinant Human Tissue Factor and Synthetic Phospholipids
Authors: Bader R , Mannucci P M M , Tripodi A , Hirsh J , Keller F , Solleder E M , Hawkins P , Peng M , Pelzer H , Teijidor L M , Ramirez I F , Kolde H-J
Published: July 2018

A new PT reagent based on recombinant human tissue factor and synthetic phospholipids (phosphatidyl choline and phosphatidyl serine) with defined fatty acid side chains was calibrated against BCT/253 and CRM 149R. A small but consistent bias in the International Sensitivity Index (ISI) value was obtained using either the human or rabbit brain reference material. ISI values were around 1.0 or slightly lower depending on the respective instrument. Mixing studies with factor deficient plasmas showed a high factor sensitivity especially for factor VII as compared to commercial rabbit brain or human placenta thromboplastin. In an international field trial the reagent was tested using fully or semi automated Electra™ coagulometers in 4 different laboratories. Results with normal samples were in excellent agreement among the different laboratories. Mean values were 10.9, 10.9, 11.0, 11,7 s with a range of 9.5 to 12.5 s. Results of males and females were not different. In patients with liver disease very similar PT activities were found as compared to sensitive rabbit brain or human placental thromboplastins. In normals and patients with oral anticoagulation INR values correlated very well against BCT (r = 0.98, regression line y =-0.07 + 0.9 x). The distribution of samples was linear over the whole range. In the comparison against sensitive rabbit brain thromboplastin or human placental thromboplastin similar correlations were found. In a few cases higher INR values were observed for the recombinant reagent especially in patients with intensive treatment. Factor assays in those patients showed at least the strong reduction of one vitamin Independent coagulation factor. Over all the linearity was better against the rabbit brain reagent than against the human placental reagent which is slightly less factor VII sensitive as shown in mixing studies with normal and factor VII deficient plasma. Precision studies in the 4 laboratories showed excellent reproducibility of lyophilised controls or local patient plasma pools for all reagents with a better performance of the recombinant reagent. C. V. values from day to day ranged from 1.3% to 5% for normal and abnormal controls.

These results show that the recombinant PT reagent, especially in conjunction with a precise automated instrument, may improve the results of PT testing and thus may lead to better patient care.
Refold and Characterization of Recombinant Tissue Factor Pathway Inhibitor Expressed in Escherichia coli
Authors: Diaz-Collier Judy A , Palmier Mark O , Kretzmer Kuniko K , Bishop Bruce F , Combs Rodney G , Obukowicz Mark G , Frazier Ronald B , Bild Gary S , Joy William D , Hill Steven R , Duffin Kevin L , Gustafson Mark E , Junger Kurt D , Grabner Roy W , Galluppi Gerald R , Wun Tze-Chein
Published: July 2018

Human tissue factor pathway inhibitor (TFPI) was expressed in E. coli as a non-glycosylated protein with an additional alanine attached to the aminoterminus of the wild type molecule. High-level expression was obtained with pMON6875, a plasmid containing a tac promoter, Gene 10 leader from bacteriophage T7, methionine-alanine-TFPI coding sequence, and the p22 transcriptional terminator. In this system, TFPI accounted for about 5-10% of the total cell protein. The inclusion bodies containing TFPI were sulfitolyzed, purified by anion-exchange chromatography, refolded through a disulfide interchange reaction, and further fractionated by Mono S cation exchange chromatography. The Mono S resin resolved a peak of highly active TFPI from relatively inactive and possibly misfolded molecules. The E. coli TFPI was shown to be about two-fold more active, on a molar basis, than full- length human SK hepatoma TFPI in a tissue factor-induced clotting assay in human plasma.
Prevention of Intra-Coronary Thrombosis in the Anaesthetised Dog: The Importance of Thromboxane A2 and Thrombin
Authors: White Brian P , Sullivan Andrew T , Lumley Philip
Published: July 2018

Vapiprost (GR32191, a TxA2 antagonist), r-hirudin, aspirin, ticlopidine and aspirin plus ticlopidine were examined for their ability to prevent electrically-induced thrombosis in an artifically stenosed coronary artery in the anaesthetised dog. Drugs or vehicle were administered prior to a 2 h period of electrical damage which was followed by a further 2 h observation period. In all vehicle-treated animals, blood flow markedly declined with onset of the damaging current; 80% completely occluded. All treatments reduced the incidence of complete occlusion to a similar extent. Vapiprost and r-hirudin also largely prevented the decline in blood flow both during and following the damage period whilst aspirin and ticlopidine, either alone or in combination were much less effective. With r-hirudin treatment, marked cyclic changes in flow occurred throughout the experiment; these were abolished by administration of vapiprost. In this dog model, TxA2 and thrombin appear to work in concert to produce coronary thrombosis, ADP being of minor importance. The superior effect ol vapiprost over aspirin suggests a beneficial role for endogenous prostacyclin.
Expression of a Paternal History of Premature Myocardial Infarction on Fibrinogen, Factor VIIC and PAI-1 in European Offspring - The EARS Study
Authors: Bara L , Nicaud V , Tiret L , Cambien F , Samama M M ,   On behalf of the EARS group
Published: July 2018

To assess the role of genetic and environmental factors in the predisposition to atherosclerosis, 682 students whose father had suffered a myocardial infarction before the age of 55 (“cases”) and 1312 controls matched for age and sex, were recruited from 14 Universities in Europe. Fibrinogen, factor Vile and PAI-1 were compared between cases and controls across European regions. Fibrinogen and factor Vile were positively correlated with BMI, smoking and contraception. PAI-1 was positively and independently correlated with BMI and waist-to-hip ratio, and negatively with contraception. Factor Vile and PAI-1 were correlated with cholesterol and triglycerides, and fibrinogen was weakly correlated with LDL-cholesterol. After adjustment for covariates and lipids, fibrinogen level was significantly higher in male cases than in controls (2.38 vs 2.29, p <0.01). No such difference was found in females (2.59 vs 2.57 - NS). There was no significant case/control difference for factor Vile and PAI-1. These results support the hypothesis that fibrinogen is a transmissible risk factor of coronary artery disease in males. No such evidence was provided for factor Vile and PAI-1.
New Immunocapture Enzyme (ICE) Assay for Quantification of Cancer Procoagulant Activity: Studies of Inhibitors
Authors: Mielicki Wojciech P , Tagawa Maureen , Gordon Stuart G
Published: July 2018

A new, sensitive and specific immunocapture enzyme (ICE) assay for quantitation of the enzymatic activity of cancer procoagulant (CP) has been developed. The assay had good reproducibility (inter- and intra-assay CV were 6.4% and 5.7% respectively) and was linear for concentrations of CP from 0.5 μg/ml to 10 μg/ml (r2 = 0.995). Using this assay the inhibition of CP by iodoacetamide, mercuric chloride, E-64, leupcptin and antipain was demonstrated. There was no significant effect of cystatin and natural plasma proteinase inhibitors α1-antitrypsin, α1-antichymotrypsin, α2-macroglobulin and antithrombin-III/heparin, on the activity of the CP.
Radiolabeled r-Hirudin as a Measure of Thrombin Activity at, or within, the Rabbit Aorta Wall In Vitro and In Vivo
Authors: Hatton Mark W C , Ross-Ouellet Bonnie
Published: July 2018

The behavior of 125I-labeled recombinant hirudin towards the uninjured and de-endothelialized rabbit aorta wall has been studied in vitro and in vivo to determine its usefulness as an indicator of thrombin activity associated with the aorta wall. Thrombin adsorbed to either sulfopropyl-Sephadex or heparin-Sepharose bound >95% of 125I-r-hirudin and the complex remained bound to the matrix. Binding of 125I-r-hirudin to the exposed aorta subendothelium (intima-media) in vitro was increased substantially if the tissue was pre-treated with thrombin; the quantity of l25I-r-hirudin bound to the de-endothelialized intima-media (i.e. balloon-injured in vitro) correlated positively with the quantity of bound 131I-thrombin (p <0.01). Aortas balloon-injured in vivo were measured for thrombin release from, and binding of 125I-r-hirudin to, the de-endothelialized intimal surface in vitro; 125I-r-hirudin binding correlated with the amount of active thrombin released (p <0.001). Uptake of 125I-r-hirudin by the aorta wall in vivo was proportional to the uptake of 131I-fibrinogen (as an indicator of thrombin activity) before and after balloon injury. After 30 min in the circulation, specific 125I-r-hirudin binding to the uninjured and de-endo- thelialized (at 1.5 h after injury) aorta wall was equivalent to 3.4 (± 2.5) and 25.6 (±18.1) fmol of thrombin/cm2 of intima-media, respectively. Possibly, only hirudin-accessible, glycosaminoglycan-bound thrombin is measured in this way.
Whole Blood Clot Lysis: Enhanced by Exposure to Autologous but not to Homologous Plasma
Authors: Beer J H , Kläy H P , Herren T , Haeberli A , Straub P W
Published: July 2018

A new phenomenon is described: Whole blood clots lyse faster in the plasma of the same donor than in another donor’s plasma. We have confirmed this finding in 68 healthy volunteers by a standardized, pairwise analysis and have found a mean difference in clot weights of 8.8 ± 0.99% (SEM, p<0.0001) after 6 h of urokinase-induced (200 U/ml) clot lysis. Nu difference was found in a group of 7 pairs of identical twins. Further analysis revealed that increasing concentrations of platelets in the plasma reduced the difference significantly but did not abolish it. A 1:1 mixture of autologous with homologous plasma reduced the autologous advantage by almost 50%, thus making an inhibitor unlikely. The absence of cellular components in clots of platelet- poor plasma resulted in the loss of the advantage after 2 h of lysis, but not in the early phase. We conclude that there is a clear advantage of autologous over homologous clot lysis. Potential mechanisms are discussed and include an increased affinity of enzymes for their substrates in a given individual.
Effect of Human IgG Antiphospholipid Antibodies on an In Vivo Thrombosis Model in Mice
Authors: Pierangeli Silvia S , Barker John H , Stikovac Dejzi , Ackerman Douglas , Anderson Gary , Barquinero Jordi , Acland Robert , Nigel Harris E
Published: July 2018

High levels of IgG antiphospholipid antibodies (aPL) have been associated with clinical thrombosis. It is uncertain however whether these antibodies play a direct role in thrombosis or are merely epiphenomena. To investigate whether antiphospholipid antibodies might play a role in thrombosis, we utilized a novel mouse model in which the dynamics of in vivo thrombosis can be studied. CD1 mice (26-30 g) were passively immunized with 25 mg of human IgG from a patient with the Antiphospholipid Syndrome (IgG-APS) (n = 17), IgG from normal pooled sera (TgG-NHS) (n = 9). or saline solution (n = 17), followed by 40 mg of the same preparations at 48 h. At 72 h, levels of human aPL antibodies, detected using the anticardiolipin ELISA test (aCL ELISA test), in mice immunized with IgG-APS, were 50-100 GPL units. Each animal was anesthetized, femoral vein minimally mobilized and subjected to a standardized “pinch” injury to induce thrombosis. The vessel was transilluminated using acrylic optical fibers connected to a light source, and clot formation and dissolution were visualized by a standard surgical microscope equipped with a video camera, video recorder, and computer assisted analysis system. Results showed that average clot size was significantly larger in mice immunized with IgG-APS compared to those treated with saline (p <0.037). In addition, the thrombus persisted longer in a significantly higher number of mice immunized with IgG-APS (10/17) compared to mice immunized with IgG-NHS (1/9) or saline (2/12) (p <0.02). These data suggest that IgG-APS may play a role in thrombus formation in humans. In addition, this study shows the feasibility of using this in vivo murine thrombus model for study of the effects of aPL antibodies.
Haemostatic Activation before and after Surgery in Patients with and without Gastric Malignancy
Authors: Rahr H B , Sørensen J V , Larsen J F , Svendsen Jensen F , Bredahl C , Nieuwenhuizen W
Published: July 2018

Pre- and postoperative plasma levels of Prothrombin Fragment 1 + 2 (F1+2), Thrombin-antithrombin III complex (TAT), Fibrinopeptide A (FpA), Fibrin and Fibrinogen Degradation Products (FbDP, FgDP) and Soluble Fibrin (SF) were measured in 40 patients undergoing gastric surgery in order to compare patients operated for benign (n = 21) and malignant (n = 19) disease. Plasma levels of F1+2, TAT, FbDP and SF on the first postoperative day were significantly higher than before operation. F1+2 and FbDP levels were further increased one week after surgery, at which time FgDP levels were also higher than preoperatively. A significant postoperative increase in FpA levels was found only in patients with malignant disease. When age was taken into consideration, significant differences between patients with and without malignancy were found only in the late postoperative period, as cancer patients had higher FpA and FbDP levels one week after surgery and higher FbDP levels one month after discharge from hospital.
Molecular Basis of Antithrombin Type I Deficiency: The First Large In-frame Deletion and Two Novel Mutations in Exon 6
Authors: Emmerich J , Chadeuf G , Alhenc-Gelas M , Gouault-Heilman M , Toulon P , Fiessinger J N , Aiach M
Published: July 2018

We report three novel mutations accounting for cases of inherited type I antithrombin (AT) deficiency. Using the polymerase chain reaction (PCR) and direct sequencing of the coding sequences of the AT gene, we found one mutation in exon 4 and two in exon 6. A deletion of 105 bp causing an in-frame deletion of 35 amino acids between Tyr 240 and Gly 276 was found in exon 4. In a second kindred, deletion of two adenines in codon 412-413 introduced a frameshift and a stop codon at position 431. The last mutation was an insertion of ACCG in codon 387, generating a frameshift with a stop codon located at the normal position.

The finding of a sequence repeat of nine residues located at the 5’and 3’ ends of the deleted fragment might explain the 105 bp deletion by slippage and mispairing at the replication fork during DNA synthesis. The second mutation is the fourth described within a region of six amino acids (between Phe 408 and Arg 413), which seems to be a cluster of mutations. In this case, the presence of a double repeat sequence - TTCCT and AACA - flanking this region could be particularly favorable for slipped mispairing.

These results confirm that human gene mutations are not random events but are strongly influenced by DNA flanking sequences.
Both Fish Oil and Olive Oil Lowered Plasma Fibrinogen in Women with High Baseline Fibrinogen Levels
Authors: Oosthuizen W , Vorster H H , Jerling J C , Barnard H C , Smuts C M , Silvis N , Kruger A , Venter C S
Published: July 2018

This double-blind, cross-over study with olive oil as placebo, examined the effect of a daily dosage of 6 g fish oil on cardiovascular risk markers of 20 healthy young volunteers (10 men, 10 women). Serum lipids and lipoproteins, and plasma coagulation and fibrinolytic enzymes, including fibrinogen concentrations and plasminogen activator inhibitor-1 (PAI-1) activity were measured at baseline and after 6-week supplementation of either fish or olive oil. The results showed that fish oil had an independent lowering effect on triglycerides and coagulation factors Vc and VIIc. Both fish and olive oil significantly raised PAI-1 levels and lowered plasma factor Xc and fibrinogen levels in the women, who had higher initial levels than the men. Mean fibrinogen levels of the women were lowered from 3.23 ± 0.98 to 2.64 ± 0.55 g/1 and from 3.19 ± 0.72 to 2.66 ± 0.49 g/1 by fish and olive oil respectively. This study raises the question whether a particular fatty acid or group of fatty acids, or another constituent of the oil such as vitamin E may be responsible for the fibrinogen lowering effect.
Characterization of a Novel Streptokinase Produced by Streptococcus equisimilis of Non-Human Origin
Authors: Nowicki Stephen T , Minning-Wenz Dena , Johnston Kenneth H , Lottenberg Richard
Published: July 2018

Streptokinases are proteins with plasminogen activator activity produced by certain hemolytic streptococci. We previously identified equine streptococcal isolates which produced streptokinases (ESKs) that bound both human and equine plasminogen but only readily activated equine plasminogen (14). This property was exploited to purify a representative ESK produced by Streptococcus equisimilis strain 87-542-W. Affinity chromatography with human plasminogen resulted in the isolation of a M~49,000 molecule with two isoforms. This ESK was subsequently compared to well characterized streptokinases (HSKs) that efficiently activate human plasminogen. Differences in streptokinases were identified in the highly conserved amino-terminal amino acid sequence, peptide maps, and antigenic properties, and these differences were supported by DNA hybridization studies. These results indicate that the family of proteins identified as streptokinases has much greater diversity than previously appreciated.
Ex Vivo Inhibition of Platelet Aggregation in Patients with Severe Limb Arteriopathy Treated with BAY U3405, a Specific IX A2 Receptor Antagonist
Authors: Bellucci S , Kedra W , Groussin H , Jaillet N , Molho-Sabatier P , Andreassian B , Tobelem G
Published: July 2018

A double-blind, placebo-controlled randomized study with BAY U3405, a specific thromboxane A2 (TX A2) receptor blocker, was performed in patients suffering from severe stade II limb arteriopathy. BAY U3405 or placebo was administered in 16 patients at 20 mg four times a day (from day 1 to day 3). Hemostatic studies were done before therapy, and on day 2 and day 3 under therapy. On day 3, BAY U3405 was shown to induce a highly statistically significant decrease of the velocity and the intensity of the aggregations mediated by arachidonic acid (56 ± 37% for the velocity, 58 ± 26% for the intensity) or by U46619 endoperoxide analogue (36 ± 35% for the velocity, 37 ± 27% for the intensity). Similar results were already observed on day 2. By contrast, such a decrease was not noticed with ADP mediated platelet aggregation. Furthermore, plasma levels of betathrombo-globulin and platelet factor 4 remained unchanged. Peripheral hemodynamic parameters were also studied. The peripheral blood flow was measured using a Doppler ultrasound; the pain free walking distance and the total walking ability distance were determined under standardized conditions on a treadmill. These last two parameters show a trend to improvement which nevertheless was not statistically significant. All together these results encourage further in vivo studies using BAY U3405 or related compounds on a long-term administration.