bluuebunny/crossref_metadata_2025 huggingface dataset:
The leaf transcriptome of fennel (Foeniculum vulgare Mill.) enables characterization of the t-anethole pathway and the discovery of microsatellites and single-nucleotide variants
Authors: Palumbo Fabio , Vannozzi Alessandro , Vitulo Nicola , Lucchin Margherita , Barcaccia Gianni
Published: July 2018

Fennel is a plant species of both agronomic and pharmaceutical interest that is characterized by a shortage of genetic and molecular data. Taking advantage of NGS technology, we sequenced and annotated the first fennel leaf transcriptome using material from four different lines and two different bioinformatic approaches: de novo and genome-guided transcriptome assembly. A reference transcriptome for assembly was produced by combining these two approaches. Among the 79,263 transcripts obtained, 47,775 were annotated using BLASTX analysis performed against the NR protein database subset with 11,853 transcripts representing putative full-length CDS. Bioinformatic analyses revealed 1,011 transcripts encoding transcription factors, mainly from the BHLH, MYB-related, C2H2, MYB, and ERF families, and 6,411 EST-SSR regions. Single-nucleotide variants of SNPs and indels were identified among the 8 samples at a frequency of 0.5 and 0.04 variants per Kb, respectively. Finally, the assembled transcripts were screened to identify genes related to the biosynthesis of t-anethole, a compound well-known for its nutraceutical and medical properties. For each of the 11 genes encoding structural enzymes in the t-anethole biosynthetic pathway, we identified at least one transcript showing a significant match. Overall, our work represents a treasure trove of information exploitable both for marker-assisted breeding and for in-depth studies on thousands of genes, including those involved in t-anethole biosynthesis.
Changing of the Genomic Pattern of Salmonella Enteritidis Strains Isolated in Brazil Over a 48 year-period revealed by Whole Genome SNP Analyses
Authors: Campioni Fábio , Cao Guojie , Kastanis George , Janies Daniel A. , Bergamini Alzira Maria Morato , Rodrigues Dália dos Prazeres , Stones Robert , Brown Eric , Allard Marc W. , Falcão Juliana Pfrimer
Published: July 2018

Salmonella Enteritidis became the main serovar isolated from gastroenteritis cases in Brazil after the 90’s. In this study we used whole genome sequence analysis to determine the phylogenetic relationships among a collection of strains isolated in Brazil to identify possible genomic differences between the strains isolated in the pre and post-epidemic period. Also, we compared our data from strains isolated in Brazil to strains available in the public domain from other South American countries. Illumina technology was used to sequence the genome of 256 Salmonella Enteritidis strains isolated over a 48 year-period in Brazil, comprising the pre- and post-epidemic period. Phylogenetic analyses revealed distinct lineages for strains isolated before and after 1994. Moreover, the phage region SE20 that may be related to the emergence of Salmonella Enteritidis worldwide was present only in strains of the post-epidemic cluster. In conclusion, our results showed that the genomic profile of Salmonella Enteritidis strains isolated in Brazil shifted after 1994, replaced by a global epidemic group of strains. It may be hypothesized that the presence of the prophage SE20 might have conferred to these strains a better ability to colonize chicken and consequently to infect and cause disease in humans, which might better explain the increase in the number of S. Enteritidis cases in Brazil and other South American countries. However, to verify this hypothesis further studies are needed.
Calorimetric and spectroscopic detection of the interaction between a diazo dye and human serum albumin
Authors: Patel Bhargav R. , Kerman Kagan
Published: July 2018

Dye effluents are one of the main causes of water pollution. In this report, calorimetric and multi-spectroscopic techniques are utilised to probe interactions of Congo red (diazo dye) with human serum albumin.
Electrospinning of in situ crosslinked recombinant human collagen peptide/chitosan nanofibers for wound healing
Authors: Deng Aipeng , Yang Yang , Du Shimei , Yang Shulin
Published: July 2018

Crosslink RHC/chitosan nanofibers in situ to simplify the preparation procedure and improve the properties of nanofibrous membranes.
High-yielding18F radiosynthesis of a novel oxytocin receptor tracer, a probe for nose-to-brain oxytocin uptakein vivo
Authors: Beard Rhiannon , Singh Nisha , Grundschober Christophe , Gee Antony D. , Tate Edward W.
Published: July 2018

Probing the mechanism of intranasal oxytocin brain uptake through generation and validation of a novel peptide PET tracer.
Mechanochemical dehydrocoupling of dimethylamine borane and hydrogenation reactions using Wilkinson's catalyst
Authors: Schumacher Christian , Crawford Deborah E. , Raguž Branimir , Glaum Robert , James Stuart L. , Bolm Carsten , Hernández José G.
Published: July 2018

Synthesis and use of the emblematic Wilkinson's catalyst by mechanochemistry was achieved.
Controlling the shape of wedge-like α-GaOOH particles formed by a hydrolysis process using sodium acetate as a growth modifier
Authors: Shiba Fumiyuki , Yuasa Minako , Okawa Yusuke
Published: July 2018

Sodium acetate preferentially suppresses the growth along thec-axis during the hydrolysis formation of α-GaOOH particles.
Surface aging investigation by means of an AFM-based methodology and the evolution of conservative nanoscale interactions
Authors: Chiesa Matteo , Lai Chia-Yun
Published: July 2018

The divergence in physical properties of surfaces exposed to airborne contaminants in the atmosphere has been widely investigated in recent years, but agreement regarding the role that airborne hydrocarbons and water contamination have on surface property evolution remains elusive.
Correction: Hybrid micro-/nano-structures derived from metal–organic frameworks: preparation and applications in energy storage and conversion
Authors: Cao Xiehong , Tan Chaoliang , Sindoro Melinda , Zhang Hua
Published: July 2018

Correction for ‘Hybrid micro-/nano-structures derived from metal–organic frameworks: preparation and applications in energy storage and conversion’ by Xiehong Cao et al., Chem. Soc. Rev., 2017, 46, 2660–2677.
On the alcoholysis of alkyl-aluminum(iii) alkoxy-NHC derivatives: reactivity of the Al-carbene Lewis pair versus Al-alkyl
Authors: Dardun Vincent , Escomel Léon , Jeanneau Erwann , Camp Clément
Published: July 2018

Alkyl-aluminum(iii) derivatives supported by a bifunctional alkoxy-NHC ligand display unusual reactivity at the carbene, yielding imidazolium-aluminate zwitterions.
Sodium stannate promoted double bond cleavage of oleic acid by hydrogen peroxide over a heterogeneous WO3catalyst
Authors: Li Xiukai , Choo Ping Syong Joel , Zhang Yugen
Published: July 2018

A sodium stannate additive notably improved the reaction efficiency of tungsten oxide catalysed oleic acid (OA) cleavage by hydrogen peroxide to produce azelaic acid and nonanoic acid.
Synergistic interplay of ionic liquid and dodecyl sulphate driving the oxidation state of polypyrrole based electrodes
Authors: Loguercio Lara F. , de Matos Carolina F. , de Oliveira Matheus C. , Marin Graciane , Khan Sherdil , Balzaretti Naira M. , Dupont Jairton , Leite Santos Marcos J. , Leite Santos Jacqueline F.
Published: July 2018

Herein, polypyrrole films are preparedviaelectrochemical deposition in a mixture of 1-n-butyl-3-methylimidazolium methanesulfonate ionic liquid and dodecy sulfate.
The anti-soiling performance of highly reflective superhydrophobic nanoparticle-textured mirrors
Authors: Jang Gyoung Gug , Smith D. Barton , List Frederick Alyious , Lee Dominc F. , Ievlev Anton V. , Collins Liam , Park Jaehyeung , Polizos Georgios
Published: July 2018

Highly transparent, superhydrophobic nanoparticle-textured coatings with engineered surface roughness significantly decrease the adhesion force of dust particles on the surface, resulting in soil and dust repellent performance.
Inexpensive NaX (X = I, Br, Cl) as a halogen donor in the practical Ag/Cu-mediated decarboxylative halogenation of aryl carboxylic acids under aerobic conditions
Authors: Fu Zhengjiang , Jiang Ligao , Zuo Qianming , Li Zhaojie , Liu Yanzhu , Wei Zhenhong , Cai Hu
Published: July 2018

Practical Ag/Cu-catalyzed decarboxylative halogenation between low-cost aryl carboxylic acids and NaX (X = I, Br, Cl) has been well established.
Closed-cell, emulsion-templated hydrogels for latent heat storage applications
Authors: Gui Haoguan , Zhang Tao , Guo Qipeng
Published: July 2018

Hydrogel polyHIPEs with closed-cell structures were prepared from o/w Pickering HIPEs, and applied in latent heat storage.
A thermo-/pH-responsive hydrogel (PNIPAM-PDMA-PAA) with diverse nanostructures and gel behaviors as a general drug carrier for drug release
Authors: Chen Yan , Gao Yuting , da Silva Lucília P. , Pirraco Rogério P. , Ma Mengdi , Yang Liming , Reis Rui L. , Chen Jie
Published: July 2018

The aim of this research was to develop thermo- and pH-responsive hydrogels based on H-bonds for the sustained release of the small-molecule model drug Methylene Blue (MB).
Crystal structure and pH-dependent allosteric regulation of human β-ureidopropionase, an enzyme involved in anticancer drug metabolism
Authors: Maurer Dirk , Lohkamp Bernhard , Krumpel Michael , Widersten Mikael , Dobritzsch Doreen
Published: July 2018

β-Ureidopropionase (βUP) catalyzes the third step of the reductive pyrimidine catabolic pathway responsible for breakdown of uracil-, thymine- and pyrimidine-based antimetabolites such as 5-fluorouracil. Nitrilase-like βUPs use a tetrad of conserved residues (Cys233, Lys196, Glu119 and Glu207) for catalysis and occur in a variety of oligomeric states. Positive co-operativity toward the substrate N-carbamoyl-β-alanine and an oligomerization-dependent mechanism of substrate activation and product inhibition have been reported for the enzymes from some species but not others. Here, the activity of recombinant human βUP is shown to be similarly regulated by substrate and product, but in a pH-dependent manner. Existing as a homodimer at pH 9, the enzyme increasingly associates to form octamers and larger oligomers with decreasing pH. Only at physiological pH is the enzyme responsive to effector binding, with N-carbamoyl-β-alanine causing association to more active higher molecular mass species, and β-alanine dissociation to inactive dimers. The parallel between the pH and ligand-induced effects suggests that protonation state changes play a crucial role in the allosteric regulation mechanism. Disruption of dimer–dimer interfaces by site-directed mutagenesis generated dimeric, inactive enzyme variants. The crystal structure of the T299C variant refined to 2.08 Å resolution revealed high structural conservation between human and fruit fly βUP, and supports the hypothesis that enzyme activation by oligomer assembly involves ordering of loop regions forming the entrance to the active site at the dimer–dimer interface, effectively positioning the catalytically important Glu207 in the active site.
Comprehensive assessment for miRNA polymorphisms in hepatocellular cancer risk: a systematic review and meta-analysis
Authors: Wang Ben-Gang , Jiang Li-Yue , Xu Qian
Published: July 2018

MiRNA polymorphisms had potential to be biomarkers for hepatocellular cancer (HCC) susceptibility. Recently, miRNA single nucleotide polymorphisms (SNPs) were reported to be associated with HCC risk, but the results were inconsistent. We performed a systematic review with a meta-analysis for the association of miRNA SNPs with HCC risk. Thirty-seven studies were included with a total of 11821 HCC patients and 15359 controls in this meta-analysis. We found hsa-mir-146a rs2910164 was associated with a decreased HCC risk in the recessive model (P=0.017, OR = 0.90, 95% confidence interval (CI) = 0.83–0.98). While hsa-mir-34b/c rs4938723 was related with an increased HCC risk in the co-dominant model (P=0.016, odds ratio (OR) = 1.19, 95%CI = 1.03–1.37). When analyzing the Hepatitis B virus (HBV)-related HCC risk, hsa-mir-196a-2 rs11614913 was associated with a decreased HBV-related HCC risk in the co-dominant and allelic models. And hsa-mir-149 rs2292832 was found to be associated with a decreased HBV-related HCC risk in the dominant and recessive models. In conclusion, hsa-mir-146a rs2910164 and hsa-mir-34b/c rs4938723 could be biomarkers for the HCC risk while hsa-mir-196a-2 rs11614913 and hsa-mir-149 rs2292832 had potential to be biomarkers for HBV-related HCC risk.
Characterization of low-mass companion HD 142527 B
Authors: Christiaens V. , Casassus S. , Absil O. , Kimeswenger S. , Gomez Gonzalez C. A. , Girard J. , Ramírez R. , Wertz O. , Zurlo A. , Wahhaj Z. , Flores C. , Salinas V. , Jordán A. , Mawet D.
Published: July 2018

Context. The circumstellar disk of the Herbig Fe starHD 142527is host to several remarkable features including a warped inner disk, a 120 au-wide annular gap, a prominent dust trap and several spiral arms. A low-mass companion,HD 142527B, was also found orbiting the primary star at ~14 au.

Aims. This study aims to better characterize this companion, which could help explain its impact on the peculiar geometry of the disk.

Method. We observed the source with VLT/SINFONI inH + Kband in pupil-tracking mode. Data were post-processed with several algorithms based on angular differential imaging (ADI).

Results.HD 142527B is conspicuously re-detected in most spectral channels, which enables us to extract the first medium-resolution spectrum of a low-mass companion within 0.″1 from its central star. Fitting our spectrum with both template and synthetic spectra suggests that the companion is a young M2.5 ± 1.0 star with an effective temperature of 3500 ± 100 K, possibly surrounded with a hot (1700 K) circum-secondary environment. Pre-main sequence evolutionary tracks provide a mass estimate of 0.34 ± 0.06M⊙, independent of the presence of a hot environment. However, the estimated stellar radius and age do depend on that assumption; we find a radius of 1.37 ± 0.05R⊙(resp. 1.96 ± 0.10R⊙) and an age of 1.8-0.5+1.2Myr (resp. 0.75 ± 0.25 Myr) in the case of the presence (resp. absence) of a hot environment contributing inH+K. Our new values for the mass and radius of the companion yield a mass accretion rate of 4.1–5.8 × 10−9M⊙yr−1(2–3% that of the primary).

Conclusions. We have constrained the physical properties ofHD 142527B, thereby illustrating the potential for SINFONI+ADI to characterize faint close-in companions. The new spectral type makesHD 142527B a twin of the well-known TW Hya T Tauri star, and the revision of its mass to higher values further supports its role in shaping the disk.
Prostate Artery Embolization: Indication, Technique and Clinical Results
Authors: Teichgräber Ulf , Aschenbach René , Diamantis Ioannis , von Rundstedt Friedrich-Carl , Grimm Marc-Oliver , Franiel Tobias
Published: July 2018

Background Prostate artery embolization (PAE) is a new embolization therapy to treat benign prostate syndrome (BPS).

Materials and Methods This review article presents the rationale and impact mechanism of PAE, criteria for patient selection, and discusses the anatomy of prostate arteries. The study results are seen in the context of complications and clinical partners.

Results Important preconditions for successful prostate artery embolization are a strict indication, precise knowledge of the anatomy of the pelvic arteries and advanced interventional-radiological skills. Several studies showed that urological parameters after prostate artery embolization improve at a similar level as for established post-surgical treatments. At the same time, it could be proven that prostate artery embolization has no impact on erectile function and is associated with a relatively low complication rate.

Conclusion PAE is increasingly developing to an alternative for the established surgical treatments in BPS patients.

Key Points 

Citation Format
Derivatives of Alkyl 2-Hydroxy-3-oxocyclopent-1-enecarboxylates and Intermolecular [4+2] Cycloadditions of Cyclopentadienones Prepared Therefrom
Authors: Harmata Michael , Garimalla Aswin
Published: July 2018

Some derivatives of alkyl 2-hydroxy-3-oxocyclopent-1-enecarboxylates have been synthesized and their reactivity as progenitors of cyclopentadienones for intermolecular [4+2]-cycloaddition reactions has been evaluated. It was found that the derivative containing a phosphate ester leaving group gave better yields in the cycloaddition reaction among the derivatives studied. The yields of the cycloaddition reactions were moderate, perhaps due side reactions not leading to the reactive intermediate cyclopentadienone.
Venous Thromboembolism and High Grade Gliomas
Authors: Dhami Mandeep S , Bona Robert D , Calogero John A , Hellman Richard M
Published: July 2018

A retrospective study was done to determine the incidence of and the risk factors predisposing to clinical venous thromboembolism (VTE) in patients treated for high grade gliomas. Medical records of 68 consecutive patients diagnosed and treated at Saint Francis Hospital and Medical Center from January 1986 to June 1991 were reviewed. The follow up was to time of death or at least 6 months (up to December 1991). All clinically suspected episodes of VTE were confirmed by objective tests. Sixteen episodes of VTE were detected in 13 patients for an overall episode rate of 23.5%. Administration of chemotherapy (p = 0.027, two tailed Fisher exact test) and presence of paresis (p = 0.031, two tailed Fisher exact test) were statistically significant risk factors for the development of VTE. Thrombotic events were more likely to occur in the paretic limb and this difference was statistically significant (p = 0.00049, chi square test, with Yates correction). No major bleeding complications were seen in the nine episodes treated with long term anticoagulation.

We conclude that venous thromboembolic complications are frequently encountered in patients being treated for high grade gliomas and the presence of paresis and the administration of chemotherapy increases the risk of such complications.
Comparison of Real-Time B-Mode Ultrasonography and Doppler Ultrasound with Contrast Venography in the Diagnosis of Venous Thrombosis in Symptomatic Outpatients
Authors: Cogo Alberto , Lensing Anthonie W A , Prandoni Paolo , Büller Harry R , Girolami Antonio , ten Cate Jan Wouter
Published: July 2018

In a prospective study, we compared real-time B-mode ultrasonography, using the simple criteria of common femoral and popliteal vein compressibility (Compression US), and Doppler ultrasound, using a standardized technique (Doppler US), with contrast venography in 158 consecutive outpatients symptomatic for deep-vein thrombosis of the lower limbs (DVT). For proximal vein thrombosis, the sensitivities documented for Compression US and Doppler US were 100% (95% CI: 90% to 100%) and 89% (95% CI: 76% to 96%), respectively. This difference is not statistically significant (p = 0.056). For all thrombi (including isolated calf-vein thrombosis), however, the sensitivity of Compression US was significantly higher than that of Doppler US (95% and 76%, respectively; p <0.04). Compression US was normal in all patients with normal venogram (specificity, 100%; 95% CI: 95% to 100%), while Doppler US was abnormal in two patients with normal venogram (specificity, 98%; 95% CI: 92% to 100%). The specificities of the two tests did not differ significantly. The results of our comparison suggest that Compression US is superior to Doppler US in the detection of DVT in symptomatic outpatients.
Functional Characterization of a Variant Prekallikrein (PK Zürich)
Authors: Wuillemin W A , Furlan M , von Felten A , Lämmle B
Published: July 2018

The plasma of a 68-year-old man with cross reacting material (CRM)-positive prekallikrein (PK) deficiency was studied. PK clotting activity was <0.01 U/ml, and PK antigen was 0.1 U/ml. No circulating anticoagulant against PK was detectable. The abnormal PK molecule, denoted as prekallikrein Zürich, was partially characterized by immunological and functional studies on the propositus’ plasma. Immunobiotting analysis showed the abnormal PK being a single chain molecule of the same M
r (80 kDa) as normal PK. Dextran sulfate activation of the propositus’ plasma did not lead to proteolytic cleavage of the variant PK molecule, in contrast to dextran sulfate activation of a mixture of 1 volume normal plasma and 9 volumes CRM-negative PK deficient plasma. Agarose gel electrophoresis followed by immunoblotting demonstrated that PK Zürich was complexed with high molecular weight kininogen similarly to PK in normal plasma. Incubation of the propositus’ plasma with purified β-FXIIa resulted in impaired cleavage of PK Zürich when compared with PK hydrolysis in a mixture of 10% normal plasma and 90% CRM-negative PK deficient plasma. Moreover, proteolytically cleaved PK Zürich showed no enzymatic activity against factor XII and high molecular weight kininogen.

These studies show that the functional defect of prekallikrein Zürich is due to an impaired cleavage by activated factor XII and probably the lack of enzymatic activity of the cleaved variant molecule.
Antigens of Tissue Plasminogen Activator and Plasminogen Activator Inhibitor 1: Correlates in Nonsmoking Japanese and Caucasian Men and Women
Authors: Iso Hiroyasu , Folsom Aaron R , Koike Kazuko A , Sato Shinichi , Wu Kenneth K , Shimamoto Takashi , Iida Minoru , Komachi Yoshio
Published: July 2018

We reported in a 1987 preliminary study that tissue plasminogen activator antigen was significantly higher in American Caucasian men than in Japanese men. To further examine possible differences in fibrinolytic activity between the two races, an expanded study was conducted in a total of 300 nonsmoking men and women aged 47-69 years in two population-based samples: rural Japanese living in Akita and Caucasians living in Minneapolis-St. Paul, MN. Antigens of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) were measured. Mean t-PA antigen was 2.3 ng/ml higher in Caucasian men than in Japanese men (P <0.001), but no race difference was seen for women (P = 0.59). Mean PAI-1 was higher in Caucasians than in Japanese for both sexes, and the race difference in mean was 1.8 ng/ml for men (P = 0.07) and 4.4 ng/ml for women (P <0.001). Both t-PA and PAI-1 were associated positively with body mass index and blood triglycerides for all sex-race groups, and positively with alcohol intake for Japanese and Caucasian men. Compared to Japanese, Caucasians of both sexes had higher levels of body mass index and blood triglycerides, and lower average intake of alcohol among men. Even when adjusted for body mass index, triglycerides, alcohol and other cardiovascular risk factors, the race difference in mean t-PA antigen persisted for men (P <0.001), as did the difference in mean PAI-1 for men (P = 0.03) and women (P = 0.001). If PAI-1 is a risk factor for coronary heart disease, a higher level in Caucasians than Japanese would correspond to the higher mortality rate from coronary heart disease in the United States than in Japan.
Tissue-Type Plasminogen Activator after Venous Occlusion in Pregnancy and Puerperium
Authors: Stegnar Mojca , Zore Andrej , Novak-Antolič Živa , Vovk Neva , Kruithof Egbert K O
Published: July 2018

Pregnancy is associated with depressed fibrinolysis as judged from the decreased fibrinolytic response to venous occlusion. In order to elucidate if this decreased response is due to an increase in plasminogen activator inhibitors 1 and 2 (PAI-1, PAI-2), and/or to decreased release of tissue-type plasminogen activator (t-PA) antigen during venous occlusion, 36 women (18 women with normal pregnancy and 18 with gestational hypertension without proteinuria) were followed during pregnancy and puerperium. In each woman a 20 min venous occlusion was performed in the second and in the third trimester of pregnancy and 3 days after delivery. The increase in t-PA antigen after venous occlusion relative to basal value was in the second trimester of pregnancy on average 3.7 fold, in the third trimester 4.4 fold, and so not reduced compared to non-pregnant women (3.7 fold increase). After delivery the increase in t-PA antigen was significantly enhanced (8.5 fold, p <0.005). The fibrinolytic response to venous occlusion measured by euglobulin and t-PA activity was significantly decreased in the third trimester compared to non-pregnant values (both p <0.005) and returned to somewhat higher (euglobulin clot lysis) or significantly higher (t-PA activity, p <0.01) values 3 days after delivery. Decreased euglobulin and t-PA activity after venous occlusion in the third trimester coincided with significant increases in basal PAI activity, PAI-1 antigen and PAI-2 antigen (2.9, 2.5 and >30 fold increase relative to non-pregnant values, respectively, all p <0.001). No significant differences in fibrinolytic variables were observed between nor-motensive and hypertensive pregnant women. It was concluded that t-PA antigen release during venous occlusion is not decreased during pregnancy and puerperium, and that decreased fibrinolytic response measured by global methods should be attributed to increased t-PA inhibitors. Gestational hypertension without proteinuria is not characterized by changes in fibrinolytic responses different from those observed in normal pregnancy.
Cloning of the cDNA Encoding Human Platelet CD36: Comparison to PCR Amplified Fragments of Monocyte, Endothelial and HEL Cells
Authors: Wyler B , Daviet L , Bortkiewicz H , Bordet J-C , McGregor J L
Published: July 2018

Glycoprotein CD36, also known as GPIIIb or GPIV, is a major platelet glycoprotein that bears the newly identified Naka alloantigen. The aim of this study was to clone platelet CD36 and investigate other forms of CD36-cDNA present in monocytes, endothelial and HEL cells. RNA from above mentioned cells were reverse transcribed (RT), using specific primers for CD36, and amplified by the polymerase chain reaction (PCR) technique. Sequencing the different amplified platelet derived cDNA fragments, spanning the whole coding and flanking regions, showed the near identity between platelet and CD36-placenta cDNA. Platelet CD36-cDNA cross-hybridized, in Southern blots, with RT-PCR amplified cDNA originating from monocytes, endothelial and HEL cells. However, monocytes showed a RT-PCR amplified cDNA fragment (561 bp) that was present in platelets and placenta but not on endothelial on HEL-cells. Northern blot analysis of platelet RNA hybridized with placenta CD36 indicated the presence of a major (1.95 kb) and a minor (0.95 kb) transcript. The 1.95 kb transcript was the only one observed on Northern blots of monocytes, endothelial and HEL cells. These results indicate that the structure of CD36 expressed in platelets is similar, with the exception of the 3’ flanking region, to that of placenta. Differences in apparent molecular weight between CD36 and CD36-like glycoproteins may be due to post-translational modifications.
Complexes of Nitric Oxide with Nucleophiles as Agents for the Controlled Biological Release of Nitric Oxide: Antiplatelet Effect
Authors: Diodati Jean G , Quyyumi Arshed A , Hussain Nabeen , Keefer Larry K
Published: July 2018

Nitric oxide (NO) inhibits platelet aggregation. Accordingly, we hypothesized that complexes of diethylamine and spermine with NO (DEA/NO and SPER/NO, respectively), two vasodilators previously shown to release NO spontaneously in aqueous solution, may also be useful antiplatelet agents. Platelet aggregation was measured in whole blood or platelet-rich plasma by impedance aggregometry after addition of collagen. In whole blood, the dose response curve for DEA/NO added 1 min before collagen was similar to that for aspirin (60% inhibition at 10-4 M), while SPER/NO and sodium nitroprusside were less potent by an order of magnitude. In platelet-rich plasma, 10-6 M DEA/NO caused 60% inhibition, while SPER/NO and sodium nitroprusside were as active only at 10-5 M; aspirin’s potency was unchanged from that in whole blood. In vivo, DEA/NO and sodium nitroprusside produced significant platelet inhibition 1 min after intravenous injection in the rabbit at 50 nmol/kg. Similar in vivo platelet inhibition was observed with SPER/NO and aspirin, but only at higher dose. The effects of DEA/NO and sodium nitroprusside were transient, lasting less than 30 min after treatment, while the activity of SPER/NO and aspirin was sustained throughout the 30 min experiment. The magnitude and duration of the antiplatelet effects of DEA/NO and SPER/NO correlate with the rates at which they release nitric oxide spontaneously in aqueous solution. Thus, NO/nucleophile complexes merit further exploration both as research tools and as potential antiplatelet agents.
Buccal Mucosa Bleeding Time Is Prolonged in Canine Models of Primary Hemostatic Disorders
Authors: Brooks Marjory , Catalfamo James
Published: July 2018

Bleeding times are reported in many studies using canine models, with a variety of techniques employed to adapt these tests for dogs. We evaluated a canine model of template bleeding time, the buccal mucosa bleeding time (BMBT), by examining the test’s sensitivity and specificity for defects of primary hemostasis. We examined thirty-five dogs having defined defects of either primary hemostasis (Types I, II, III von Willebrand’s disease, thrombasthenia, thrombopathia) or secondary hemostasis (hemophilia A and B, Factor VII deficiency). Comparisons of BMBT and cuticle bleeding time were made in a subset of these dogs.

All dogs having primary hemostatic disorders had long BMBT, and all factor deficient dogs had BMBT within normal range. The BMBT in canine models appears to be a specific and sensitive test of primary hemostasis; suitable for evaluating factors affecting template bleeding time and potential efficacy and thrombogenicity of treatment regimens.
A Hexagonal (II) Phase Phospholipid Neutralization Assay for Lupus Anticoagulant Identification
Authors: Triplett Douglas A , Barna Linda K , Unger Gail A
Published: July 2018

Lupus anticoagulants (LAs) are immunoglobulins (IgG, IgM, or both) which interfere with in vitro phospholipid (PL) dependent tests of coagulation (e.g. APTT, dilute PT, dilute Russell Viper Venom Time). These antibodies may be identified in a wide variety of clinical settings. With the exception of heparinized patient samples, the presence of LAs is often the most common cause of an unexplained APTT in a routine clinical laboratory. The diagnosis of LAs is difficult due to variable screening reagent sensitivity and intrinsic heterogeneity of LAs. Recently, Rauch and colleagues have shown human monoclonal hybridoma LAs were inhibited by hexagonal (II) phase PLs. In contrast, lamellar phase PLs had no effect. We have evaluated a new assay system, Staclot LA®, which utilizes a hexagonal (II) phase PL (egg phosphatidylethanolamine [EPE]) as a confirmatory test for LAs. Plasma samples from the following patient populations were studied: LA positive, heparinized, oral anticoagulated, hemophilia A and B, and specific factor inhibitors (factors V, VIII, IX). Unlike previous studies, the LA positive patients were a mixed population including: autoimmune diseases, drug-induced, and post-infection. Our findings confirm the specificity of hexagonal (II) phase PL neutralization of LAs.
Effect of Various Antiplatelet Agents on Acute Arterial Thrombosis in the Rat
Authors: Bernat A , Mares A M , Defreyn G , Maffrand J P , Herbert J M
Published: July 2018

Electrical stimulation of the rat carotid artery causes a deep medial injury and the formation of a platelet-rich thrombus.

Occlusive thrombosis at sites of vessel wall injury was significantly reduced after the oral administration of clopidogrel, a potent analogue of ticlopidine, which showed dose-dependent inhibition of the thrombus formation (ED50 = 1.0 ±0.2 mg/kg, p.o.). Accumulation of thrombotic material was also considerably reduced after the i. v. administration of SR 27417, a highly potent and selective platelet activating factor receptor antagonist (ED50 = μg/kg, i.v.), nafagrel, a thromboxane A2 synthetase inhibitor (ED50 = 1.3 mg/kg, i.v.) and hirudin (ED50 = 140 μg/ kg, i.v.). A high dose (20 mg/kg, i.v.) of the anti-adhesive tetrapeptide Arg-Gly-Asp-Ser (RGDS) showed only a slight effect on thrombus formation whereas aspirin was ineffective.

These results confirm that ADP and thromboxane A2 play key roles in the initiation and progression of arterial thrombus formation and suggest that platelet activating factor may also modulate thrombosis in this experimental model.
Multicenter Evaluation of Commercially Available Methods for the Immunological Determination of Plasminogen Activator Inhibitor-1 (PAI-1)
Authors: Declerck P J , Moreau H , Jespersen J , Gram J , Kluft C
Published: July 2018

In order to evaluate the comparability of data obtained with various available kits for the immunological determination of PAI-1 antigen in plasma and in order to investigate the underlying cause of observed differences, e. g. problems of specificity or of proper calibration of the provided standard, a multicenter study was organised in the framework of the Subcommittee of Fibrinolysis of the Scientific and Standardization Committee.

Eight different plasma samples were distributed among 16 laboratories: a pooled normal plasma, NIBSC 87/512, PAI-1 antigen depleted plasma, PAI-1 depleted plasma supplemented with 59 ng/ml active PAI-1 and four different individual plasma samples. A considerable variation in absolute values is observed between the various kits, e.g. in pooled normal plasma a value is found ranging between 7.4 and 28 ng/ml. Harmonization of all data relative to the PAI-l-depleted plasma supplemented with an exact amount of active PAI-1 (59 ng/ml), followed by a statistical analysis using a two way analysis of variance, revealed that 6 out of 7 kits yielded values that were not significantly different with coefficients of variation around 30%. Correlations between the values obtained with these kits yielded slopes between 0.75 and 1.44 with correlation coefficients between 0.973 and 0.999. Values obtained with one kit appeared to be significantly different (even after harmonization) from the other kits (p <0.001 to p <0.05). Comparison of PAI-1 antigen with the PAI activity values in the analysed samples suggests that one kit may deal with a problem of a difference in reactivity between active and latent PAI-1.

In conclusion 6 different kits yield results that, only after harmonization, are comparable and do correlate very well. Thus proper calibration of the provided standards may solve the majority of the problems observed with PAI-1 antigen kits. Differential specificity towards different conformational forms of PAI-1, as a consequence of the use of different monoclonal antibodies, is a minor, but a potential problem, that needs to be considered when comparing data obtained with various methods.
Interaction of Plasminogen Activators and Plasminogen with Heparin: Effect of Ionic Strength
Authors: Rijken Dingeman C , de Munk Gerard A W , Jie Annie F H
Published: July 2018

In order to define the possible effects of heparin on the fibrinolytic system under physiological conditions, we studied the interactions of this drug with plasminogen and its activators at various ionic strengths. As reported in recent literature, heparin stimulated the activation of Lys-plasminogen by high molecular weight (HMW) and low molecular weight (LMW) two-chain urokinase-type plasminogen activator (u-PA) and two-chain tissue-type plasminogen activator (t-PA) 10- to 17-fold. Our results showed, however, that this stimulation only occurred at low ionic strength and was negligible at a physiological salt concentration. Direct binding studies were performed using heparin-agarose column chromatography. The interaction between heparin and Lys-plasminogen appeared to be salt sensitive, which explains at least in part why heparin did not stimulate plasminogen activation at 0.15 M NaCl. The binding of u-PA and t-PA to heparinagarose was less salt sensitive. Results were consistent with heparin binding sites on both LMW u-PA and the amino-terminal part of HMW u-PA. Single-chain t-PA bound more avidly than two-chain t-PA. The interactions between heparin and plasminogen activators can occur under physiological conditions and may modulate the fibrinolytic system.
Synoviorthesis Versus Synovectomy in the Treatment of Recurrent Haemophilic Haemarthrosis: Long-Term Evaluation
Authors: Gamba G , Grignani G , Ascari E
Published: July 2018

The authors report a long-term evaluation of the effects of chemical and surgical synovectomies performed in two groups of patients which were comparable as regards number of haemarthrosis per year and joint mobility. The results obtained indicated that both synovectomy and synoviorthesis were effective in reducing the bleeding episodes; however, while synovectomy stopped almost completely the recurrence of haemarthrosis, synoviorthesis in 3 patients younger than 12 years did not modify it and in one case it even increased the bleeding tendency.

Moreover, the occurrence of arthritic pain was observed in 6 knees treated by synoviorthesis and only in 3 knees treated by synovectomy. Finally, the joint function was significantly reduced in the synoviorthesis group as compared to the synovectomy group.

In conclusion, synovectomy gave better results than synoviorthesis but the latter treatment has a number of economical and technical advantages and should therefore be considered in patients older than 12 years of age with good joint function and without evident radiological changes, in whom fulldose replacement therapy has had no effect on the recurrence of haemarthrosis.
Effect of Aspirin and Dipyridamole on the Interaction of Human Platelets with Sub-Endothelium: Studies Using Citrated and Native Blood
Authors: Weiss Harvey J , Turitto Vincent T , Vicic William J , Baumgartner Hans R
Published: July 2018

The effect of aspirin and dipyridamole ingestion on the interaction of platelets with the subendothelium was studied using both citrated blood and directly sampled (native) blood. After obtained control studies, normal human subjects ingested 0.6 g of aspirin, 150 mg of dipyridamole, or a placebo and studies were repeated 1½ hrs later. Subjects continued on placebo, aspirin (0.6 g b.i.d.) or dipyridamole (100 mg q.i.d.) for 6 days and studies were obtained 1½ hrs after the last dose. Blood was circulated through an annular chamber on whose inner core were mounted everted segments of de-endothelialized rabbit aorta. The wall shear rate was 2,600 sec-1. Surface coverage with adherent platelets and platelet thrombi, as well as several parameters of thrombus dimensions, were evaluated morphometrically. Aspirin ingestion markedly reduced platelet thrombi in citrated blood, – but had a much lesser inhibitory effect in native blood. Platelet adhesion was unaffected in native blood; it was slightly decreased in citrated blood, in contrast to previous findings in which a lower shear rate (800 sec-1) was used. Ingestion of dipyridamole did not inhibit platelet adhesion or thrombi in either citrated or native blood. The studies indicate that, with the flow conditions used, aspirin is a relatively weak inhibitor of platelet thrombus formation in directly sampled human blood.
Heparin Excretion in Intact and Hepatectomized Rats
Authors: Losito Ray , Gattiker Harry , Bilodeau Ginette
Published: July 2018

Metabolism and kinetics of 3H-heparin were compared in intact and hepatectomized rats. Rats were divided into three groups: 1) intact rats with biliary fistulas and cystostomies 2) intact rats with only cystostomies and 3) hepatectomized rats with cystostomies. Radioactivity in blood, bile and urine besides anticoagulant activity in blood and urine were examined. In addition, column chromatography of urine was used to isolate possible metabolites. Seventy percent and 80% of the radioactive dose was found in the urine of intact rats at 24 hr and 48 hr. Close to 5% of the radioactivity was found in bile or rats with a biliary fistula after 48 hr. The APTT declined to near normal values at 1 hr whether rats had a biliary fistula or not. In contrast, only 25 % of the radioactivity could be excreted into the urine of hepatectomized rats in 24 hr; the APTT did not decline as fast and at 5 hr, it was still 100 seconds. Only one radioactive component could be isolated on chromatography from all urines of these animals and appears to be similar to the original heparin. Thus, the liver has an important role to play in regulating the anticoagulant effects and excretion of heparin.
Binding of 99mTc Plasminogen on Fibrin
Authors: Juhan-Vague I , Calas M F , Roux F , Juhan C , Durand-Dessemon F , de Laforte C , Serradimigni A
Published: July 2018

Binding of plasminogen to fibrin was studied in vitro and in vivo using 99mTc Glu- and Lys-plasminogen.

Binding of Glu-plasminogen on the clot was not observed in vitro, and in vivo in the dog.

Conversely, the binding of Lys-plasminogen to fibrin displays a linear relationship to the concentration of Lys-plasminogen, up to doses exceeding equimolarity; thus suggesting the existence of several Lys-plasminogen binding sites on fibrin. Binding levels were identical, regardless of whether plasminogen was incubated in normal plasma or in plasma devoid of antiplasmin. In the dog, Lys-plasminogen bound specifically to the clot, however, clot sites could not be localized by scintigraphy in the dog or in man.
Inhibition of Human Platelet Functions by Verapamil
Authors: Ikeda Y , Kikuchi M , Toyama K , Watanabe K , Ando Y
Published: July 2018

The effects of verapamil, a coronary vasodilator, on platelet functions were studied.

Platelet aggregation induced by ADP, epinephrine or collagen was inhibited by verapamil in vitro. Calcium ionophore A23187-induced platelet aggregation was also inhibited by verapamil in a concentration dependent manner. In washed platelets, verapamil caused a dose-dependent inhibition of serotonin release induced either by thrombin or A23187 in the absence of extracellular calcium. Addition of 1 mM CaCl2 with A23187 or thrombin partially overcame this inhibition. Addition of 1 mM CaCl2 in the absence of verapamil had no effect on thrombin- or A23187-induced secretion. When verapamil was administered to the healthy volunteers at the dosage commonly used, inhibition of platelet aggregation was observed 2 hrs after the drug ingestion. It is of great interest that verapamil potentiated the anti-aggregating activity of prostacyclin in vitro.

Our results may suggest a potential role for verapamil in the treatment of thrombotic disorders.
Paris I Dysfibrinogenemia: A Point Mutation in Intron 8 Results in Insertion of a 15 Amino Acid Sequence in the Fibrinogen γ-Chain
Authors: Rosenberg Jonathan B , Newman Peter J , Mosesson Michael W , Guillin Marie-Claude , Amrani David L
Published: July 2018

Paris I dysfibrinogenemia results in the production of a fibrinogen molecule containing a functionally abnormal γ-chain. We determined the basis of the molecular defect using polymerase chain reaction (PCR) to amplify the γ-chain region of the Paris I subject’s genomic DNA. Comparative sequence analysis of cloned PCR segments of normal and Paris I genomic DNA revealed only an A→G point mutation occurring at nucleotide position 6588 within intron 8 of the Paris I γ-chain gene. We examined six normal individuals and found only normal sequence in this region, indicating that this change is not likely to represent a normal polymorphism. This nucleotide change leads to a 45 bp fragment being inserted between exons 8 and 9 in the mature γparis I chain mRNA, and encodes a 15 amino acid insert after γ350 [M-C-G-E-A-L-P-M-L-K-D-P-C-Y]. Alternative splicing of this region from intron 8 into the mature Paris I γ-chain mRNA also results after translation into a substitution of S for G at position γ351. Biochemical studies of 14C-iodoacetamide incorporation into disulfide-reduced Paris I and normal fibrinogen corroborated the molecular biologic predictions that two additional cysteine residues exist within the γpariS I chain. We conclude that the insertion of this amino acid sequence leads to a conformationallyaltered, and dysfunctional γ-chain in Paris I fibrinogen.
The Effects of Heparin and Annexin V on Fibrin Accretion after Injury in the Jugular Veins of Rabbits
Authors: Van Ryn-McKenna J , Merk H , Müller T H , Buchanan M R , Eisert W G
Published: July 2018

We compared the relative abilities of unfractionated heparin and annexin V to prevent fibrin accretion onto injured jugular veins in vivo. Heparin was used to accelerate the inhibition of thrombin by antithrombin III, and annexin V was used to inhibit the assembly of the prothrombinase complex on phospholipid surfaces, thereby blocking thrombin generation. Rabbit jugular veins were isolated in situ, a 2 cm segment was injured by perfusing it with air, and then blood flow was re-established. Five minutes later, each rabbit was injected with heparin (20 U/kg) or annexin V (0.3 mg/kg) and then with 125I-fibrinogen. The amount of 125I-fibrin accumulation onto each injured vessel wall segment was measured 4 h later. Each injured vessel was completely deendothelialized as a result of the air perfusion as demonstrated by electron microscopy. 125I-fibrin accretion onto the injured jugular veins was enhanced 2.4-fold as compared to the uninjured veins in sham-operated animals. Heparin treatment did not reduce fibrin accretion, whereas, annexin V treatment decreased fibrin accretion by 60%, p <0.05. This latter effect was achieved without sustained circulating anticoagulation. Additional experiments confirmed that the inhibitory effect of annexin V on fibrin accretion was associated with a surface specific effect, since more annexin V bound to the injured jugular vein segments as compared to the non-injured jugular veins. We conclude that, i) mild vessel wall injury (selective de-endothelialization) in veins results in a thrombogenic vessel wall; ii) the thrombogenecity of which is not inhibited by prophylactic doses of heparin; but iii) is inhibited by annexin V, which binds to injured vessel wall surface, and inhibits thrombin generation independently of antithrombin III.