The Asp272-Glu282 Region of Platelet Glycoprotein Ib Interacts with the Heparin-binding Site of -Thrombin and Protects the Enzyme from the Heparin-catalyzed Inhibition by Antithrombin III

Platelet glycoprotein Ib (GpIb) mediates interaction with both von Willebrand factor and thrombin. Thrombin binds to GpIb via its heparin-binding site (HBS) (De Candia, E., De Cristofaro, R., De Marco, L., Mazzucato, M., Picozzi, M., and Landolfi, R. (1997) Thromb. Haemostasis 77, 735–740; De Cristofaro, R., De Candia, E., Croce, G., Morosetti, R., and Landolfi, R. (1998) Biochem. J. 332, 643–650). To identify the thrombin-binding domain on GpIbα, we examined the effect of GpIbα1–282, a GpIbα fragment released by the cobra venom mocarhagin on the heparin-catalyzed rate of thrombin inhibition by antithrombin III (AT). GpIbα1–282 inhibited the reaction in a dose-dependent and competitive fashion. In contrast, the GpIbα1–271 fragment, produced by exposing GpIbα1–282 to carboxypeptidase Y, had no effect on thrombin inhibition by the heparin-AT complex. Measurements of the apparent equilibrium constant of the GpIbα1–282 binding to thrombin as a function of different salts (NaCl and tetramethyl-ammonium chloride) concentration (0.1–0.2 M) indicated a large salt dependence (Γ± = −4.5), similar to that pertaining to the heparin binding to thrombin. The importance of thrombin HBS in its interaction with GpIbα was confirmed using DNA aptamers, which specifically bind to either HBS (HD22) or the fibrinogen recognition site of thrombin (HD1). HD22, but not HD1, inhibited thrombin binding to GpIbα1–282. Furthermore, the proteolytic derivative γT-thrombin, which lacks the fibrinogen recognition site, binds to GpIbα via its intact HBS in a reaction that is inhibited by HD22. Neither α- nor γT-thrombin bound to GpIbα1–271, suggesting that the Asp272–Glu282 region of GpIbα may act as a “heparin-like” ligand for the thrombin HBS, thereby inhibiting heparin binding to thrombin. It was also demonstrated that intact platelets may dose-dependently inhibit the heparin-catalyzed thrombin inhibition by AT at enzyme concentrations <5 nM. Altogether, these findings show that thrombin HBS binds to the region of GpIbα involving the Asp272–Glu282 segment, protecting the enzyme from the inactivation by the heparin-AT system

Fibrinogen-elongated Chain Inhibits Thrombin-induced Platelet Response, Hindering the Interaction with Different Receptors

The expression of the elongated fibrinogen γ chain, termed γ′, derives from alternative splicing of mRNA and causes an insertion sequence of 20 amino acids. This insertion domain interacts with the anion-binding exosite (ABE)-II of thrombin. This study investigated whether and how γ′ chain binding to ABE-II affects thrombin interaction with its platelet receptors, i.e. glycoprotein Ibα (GpIbα), protease-activated receptor (PAR) 1, and PAR4. Both synthetic γ′ peptide and fibrinogen fragment D*, containing the elongated γ′ chain, inhibited thrombin-induced platelet aggregation up to 70%, with IC50 values of 42 ± 3.5 and 0.47 ± 0.03 μm, respectively. Solid-phase binding and spectrofluorimetric assays showed that both fragment D* and the synthetic γ′ peptide specifically bind to thrombin ABE-II and competitively inhibit the thrombin binding to GpIbα with a mean Ki ≈ 0.5 and ≈35 μm, respectively. Both these γ′ chain-containing ligands allosterically inhibited thrombin cleavage of a synthetic PAR1 peptide, of native PAR1 molecules on intact platelets, and of the synthetic chromogenic peptide d-Phe-pipecolyl-Arg-p-nitroanilide. PAR4 cleavage was unaffected. In summary, fibrinogen γ′ chain binds with high affinity to thrombin and inhibits with combined mechanisms the platelet response to thrombin. Thus, its variations in vivo may affect the hemostatic balance in arterial circulation

An Exploratory Study of User Perceptions of Payment Methods in the UK and the US

This paper presents the design and the results of a cross-cultural study of user perceptions and attitudes toward electronic payment methods. We conduct a series of semi-structured interviews involving forty participants (20 in London, UK, and 20 in Manhattan, KS, USA) to explore how individuals use the mechanisms available to them within their routine payment and banking activities. We also study their comprehension of payment processes, the perceived effort and impact of using different methods, as well as direct or indirect recollections of (suspected or actual) fraud and related interactions with banks and retailers. By comparing UK and US participants, we also elicit commonalities and differences that may help better understand, if not predict, attitudes of US customers once technologies like Chip-and-PIN are rolled out – for instance, several US participants were confused by how to use it, while UK participants found it convenient. Our results show that purchasing habits as well as the availability of rewards schemes are primary criteria influencing choices relating to payment technologies, and that inconsistencies, glitches, and other difficulties with newer technologies generate frustration sometimes leading to complete avoidance of new payment methods

Biological activity of metabolites extracted from Citrus spp. on Ceratitis capitata (Wiedemann) (Diptera: Tephritidae)

The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), is one of the most injurious pest at global level. During the last years, several electrophysiological and behavioural studies have been carried out in order to investigate plant volatile compound-insect interactions with the aim to use this knowledge in sustainable control techniques. It has been observed that lemons are not attacked by medfly, probably because of the peel oil, that is toxic to other fruit flies. In the present paper electrophysiological recordings were conducted to evaluate the insect sensitivity to peel extract and peel oil of two Sicilian cultivars (Interdonato and Lunario) of Citrus x limon (L.) Burm.f. on C. capitata females. Behavioural bioassays were also performed to show their possible biological activity (repellent, antioviposition, insecticidal). C. limon peel extracts in different solvents (petroleum ether, dichloromethane and methanol) were investigated at various concentrations using a single cell recording technique (stimulation of tarsal taste chemosensilla). Different tarsal taste cell responses to the two cultivars were recorded. The higher sensitivity was evoked by C. limon Interdonato, particularly to the methanol extract, which elicited significant increases in the spike frequency at increasing concentrations. The peel oil of the same cultivars as well as that ones of other two C. limon varieties (Monachello and Femminello) have been tested by EAG techniques. The EAG data showed a high sensitivity (about -8.0/8.5mV) of the medfly antennae to the oils of Citrus spp. and a clear dose-response relationship. Responses of adult females (virgin and mated) to Citrus spp. peel extract were quantified in a double-choice test using yellow spheres (diameter 7.0cm) housed in field cages. Preliminary tests conducted on three extracts of C. limon Interdonato and Lunario have provided interesting results. It was recorded a general decrease of the oviposition on treated spheres compared to control and in the case of the cultivar Lunario, a mortality of insects

First data on the repellent activity of essential oils of Citrus limon towards medfly (Ceratitis capitata)

The biological activity of peel essential oils of two Sicilian cultivars of C. limon, ‘Interdonato’ and ‘Lunario’, was investigated by electrophysiological recordings (EAG) and field trials on Ceratitis capitata (Wiedemann) (Diptera Tephritidae). The EAG data showed a remarkable dose-response relationship and a low activation threshold dose (10-3M) for both essential oil extracts. In field tests, essential oils showed a good repellent and antiovipositional activi- ty on ‘Navelina’ oranges, and in particular the essential oils of ‘Lunario’ pro- vided the same results of kaolin. Potential applications of essential oils from C. limon cultivars against C. capitata in organic farming are discussed

When information about one’s counterpart matters:Prevention focus increases the impact of counterpart cues on negotiation behavior

Purpose: To avoid (costly) conflict, it is imperative to uncover when negotiators cooperate. The previous study has shown that negotiators’ cooperative or competitive behavior is oftentimes guided by cues about their counterpart; information about his/her traits or behavior. Using regulatory focus theory, this paper aims to investigate when this is likely to happen. The authors hypothesize and test that because prevention focus (rather than promotion focus) is associated with concerns for safety and concrete surroundings, it strengthens the impact of counterpart cues. Design/methodology/approach: The authors used two scenario studies and one behavioral negotiation study to test the general hypothesis. The authors measured or manipulated participants’ regulatory focus, manipulated counterpart cues by varying the information negotiators received about their counterpart’s traits and behavior, and measured participants’ cooperative or competitive concession making behavior. Findings: Results from the studies confirmed that under prevention focus, negotiators’ cooperative behavior depended on whether they received cooperative versus competitive counterpart cues more than under promotion focus. Furthermore, results also showed that under prevention focus, negotiators’ behavior was relatively unaffected by their own social motivation – i.e. their personal goal to obtain favorable outcomes for oneself or for both negotiation parties. Originality/value: By showing that regulatory focus determines when counterpart cues affect negotiation behavior, this paper furthers the understanding of when contextual factors affect negotiators' behavior. In addition, it contributes to the understanding of the complex effects of prevention focus in interpersonal behavior

Soros, Child Sacrifices, and {5G}: {U}nderstanding the Spread of Conspiracy Theories on {Web} Communities

This paper presents a multi-platform computational pipeline geared to identify social media posts discussing (known) conspiracy theories. We use 189 conspiracy claims collected by Snopes, and find 66k posts and 277k comments on Reddit, and 379k tweets discussing them. Then, we study how conspiracies are discussed on different Web communities and which ones are particularly influential in driving the discussion about them. Our analysis sheds light on how conspiracy theories are discussed and spread online, while highlighting multiple challenges in mitigating them

"Is it a {Qoincidence}?": {A} First Step Towards Understanding and Characterizing the {QAnon} Movement on {Voat.co}

Online fringe communities offer fertile grounds for users to seek and share paranoid ideas fueling suspicion of mainstream news, and outright conspiracy theories. Among these, the QAnon conspiracy theory has emerged in 2017 on 4chan, broadly supporting the idea that powerful politicians, aristocrats, and celebrities are closely engaged in a global pedophile ring. At the same time, governments are thought to be controlled by "puppet masters," as democratically elected officials serve as a fake showroom of democracy. In this paper, we provide an empirical exploratory analysis of the QAnon community on Voat.co, a Reddit-esque news aggregator, which has recently captured the interest of the press for its toxicity and for providing a platform to QAnon followers. More precisely, we analyze a large dataset from /v/GreatAwakening, the most popular QAnon-related subverse (the Voat equivalent of a subreddit) to characterize activity and user engagement. To further understand the discourse around QAnon, we study the most popular named entities mentioned in the posts, along with the most prominent topics of discussion, which focus on US politics, Donald Trump, and world events. We also use word2vec models to identify narratives around QAnon-specific keywords, and our graph visualization shows that some of QAnon-related ones are closely related to those from the Pizzagate conspiracy theory and "drops" by "Q." Finally, we analyze content toxicity, finding that discussions on /v/GreatAwakening are less toxic than in the broad Voat community

Raiders of the Lost Kek: 3.5 Years of Augmented 4chan Posts from the Politically Incorrect Board

This paper presents a dataset with over 3.3M threads and 134.5M posts from the Politically Incorrect board (/pol/) of the imageboard forum 4chan, posted over a period of almost 3.5 years (June 2016-November 2019). To the best of our knowledge, this represents the largest publicly available 4chan dataset, providing the community with an archive of posts that have been permanently deleted from 4chan and are otherwise inaccessible. We augment the data with a set of additional labels, including toxicity scores and the named entities mentioned in each post. We also present a statistical analysis of the dataset, providing an overview of what researchers interested in using it can expect, as well as a simple content analysis, shedding light on the most prominent discussion topics, the most popular entities mentioned, and the toxicity level of each post. Overall, we are confident that our work will motivate and assist researchers in studying and understanding 4chan, as well as its role on the greater Web. For instance, we hope this dataset may be used for cross-platform studies of social media, as well as being useful for other types of research like natural language processing. Finally, our dataset can assist qualitative work focusing on in-depth case studies of specific narratives, events, or social theories

Binding of thrombin to glycoprotein Ib accelerates the hydrolysis of Par-1 on intact platelets

The activation of human platelets by α-thrombin is mediated at least in part by cleavage of protease-activated G-protein-coupled receptors, PAR-1 and PAR-4. Platelet glycoprotein Ibα also has a high affinity binding site for α-thrombin, and this interaction contributes to platelet activation through a still unknown mechanism. In the present study the hypothesis that GpIbα may contribute to platelet activation by modulating the hydrolysis of PAR-1 on the platelet membrane was investigated. Gel-filtered platelets from normal individuals were stimulated by α-thrombin, and the kinetics of PAR-1 hydrolysis by enzyme was followed with flow cytometry using an anti-PAR-1 monoclonal antibody (SPAN 12) that recognizes only intact PAR-1 molecules. This strategy allowed measurement of the apparentk cat/K m value for thrombin hydrolysis of PAR-1 on intact platelets, which was equal to 1.5 ± 0.1 × 107 m −1sec−1. The hydrolysis rate of PAR-1 by thrombin was measured under conditions in which thrombin binding to GpIb was inhibited by different strategies, with the following results. 1) Elimination of GpIbα on platelet membranes by mocarhagin treatment reduced the k cat/K m value by about 6-fold. 2) A monoclonal anti-GpIb antibody reduced the apparent k cat/K m value by about 5-fold. 3) An oligonucleotide DNA aptamer, HD22, which binds to the thrombin heparin-binding site (HBS) and inhibits thrombin interaction with GpIbα, reduced the apparentk cat/K m value by about 5-fold. 4) Displacement of α-thrombin from the binding site on GpIb using PPACK-thrombin reduced the apparentk cat/K m value by about 5-fold, and 5) mutation at the HBS of thrombin (R98A) caused a 5-fold reduction of the apparentk cat/K m value of PAR-1 hydrolysis. Altogether these results show that thrombin interaction with GpIb enhances the specificity of thrombin cleavage of PAR-1 on intact platelets, suggesting that GpIb may function as a “cofactor” for PAR-1 activation by thrombin