Primal Role Research Articles

Electrophysiological Activity Associated with a Cross-Modal Anapaest Rhythm: Evidence for the Vestibular Syncopation Hypothesis

Abstract We report an experiment that tested the vestibular syncopation rhythm hypothesis, which holds that the rhythmic effect of syncopation is a form of vestibular reflexive/automated response to a postural perturbation, for example during locomotion. Electrophysiological signals were recorded from the cerebral cortex and cerebellum during processing of rhythmic sequences in a sample of experienced participants. Recordings were made using four different stimulus modalities, auditory, axial, vestibular and visual, under different rhythmic timing conditions, irregular, regular and syncopated/uncertain. Brain current activity was measured using a model with10-dipole source regions of interest in each of the participants, each modality, each timing condition, and for each beat within the bar of the rhythm. The cross-modal spectral power in the frontal electroencephalogram (EEG) and the cerebellar electrocerebellogram (ECeG) was also analysed. The results show that the brain activity increases from the irregular to the regular and then from the regular to the uncertain timing conditions. However, the vestibular modality induces the greatest total brain activity across the regions of interest and exhibits the highest sensitivity to the interaction of beat structure with the timing conditions in both source currents and spectral power. These data provide further evidence to support the primal role of the vestibular system in human processing of rhythm.

β-Catenin Elicits Drp1-Mediated Mitochondrial Fission Activating the Pro-Apoptotic Caspase-1/IL-1β Signalosome in Aeromonas hydrophila-Infected Zebrafish Macrophages.

Canonical Wnt signaling plays a major role in regulating microbial pathogenesis. However, to date, its involvement in A. hydrophila infection is not well known. Using zebrafish (Danio rerio) kidney macrophages (ZKM), we report that A. hydrophila infection upregulates wnt2, wnt3a, fzd5, lrp6, and β-catenin (ctnnb1) expression, coinciding with the decreased expression of gsk3b and axin. Additionally, increased nuclear β-catenin protein accumulation was observed in infected ZKM, thereby suggesting the activation of canonical Wnt signaling in A. hydrophila infection. Our studies with the β-catenin specific inhibitor JW67 demonstrated β-catenin to be pro-apoptotic, which initiates the apoptosis of A. hydrophila-infected ZKM. β-catenin induces NADPH oxidase (NOX)-mediated ROS production, which orchestrates sustained mitochondrial ROS (mtROS) generation in the infected ZKM. Elevated mtROS favors the dissipation of the mitochondrial membrane potential (ΔΨm) and downstream Drp1-mediated mitochondrial fission, leading to cytochrome c release. We also report that β-catenin-induced mitochondrial fission is an upstream regulator of the caspase-1/IL-1β signalosome, which triggers the caspase-3 mediated apoptosis of the ZKM as well as A. hydrophila clearance. This is the first study suggesting a host-centric role of canonical Wnt signaling pathway in A. hydrophila pathogenesis wherein β-catenin plays a primal role in activating the mitochondrial fission machinery, which actively promotes ZKM apoptosis and helps in containing the bacteria.

The Malay Historical Urban Landscape: A Trajectory of the Bujang Valley Site to Jugra, Selangor

Malaysia is rich in heritage, primarily archaeological sites, historical buildings, monuments, artifacts, tangible, and intangible traditional and cultural heritage elements. Archaeotourism is one of the heritage tourism activities which is growing rapidly and attracts many visitors to the Malaysian tourism industry. In alluring visitors to the destination, an archaeological site must offer an extraordinary experiential and continually to develop meanings as an attraction to increase the numbers of visitors. The issue of historical urban landscape in Nusantara has been defined for ‘colonial’-based cities, but not pre-colonial sites. The primal and axial role of rivers, mountains and mounds in Malay history reflect not only its topographical and physical axis, but resonates from a time when the mountain claimed a primal role in the cosmology and cosmography of place. This paper attempts to find similarities between the early kingdom of Malay Peninsula, Bujang Valley archaeology site, 16th century Kota Melaka and 19th century Jugra, which reflect the historic landscape features and its uniqueness of architecture, with discovery of the remains archaeology artefact with another site in Selangor. Jugra site which similarly has the urban landscape consisting of mountain, river and key public structures. The urban form configuration and the natural setting identifies a continuity and model between a pre-Islamic and colonial-era model with similarity on the topographical, physical and historical attributes of each cultural and heritage element. This study emphasizes that by determining the historical urban landscape perspective in heritage sites, including natural physical landform, architectural archetypes, and significance in multicultural historical values, its key elements can be extracted for developing national identity and promoting heritage sites in the Malaysian tourism industry for its intangible intellectual, emotional, and aesthetical reasons.

Barrier analysis for carbon regulatory environmental policies implementation in manufacturing supply chains to achieve zero carbon

Many pressing challenges that perennially affect countries today are those related to environmental sustainability. As major entities, business organizations play a primal role in boosting the economy, stakeholders are increasingly under pressure to achieve net zero emissions. However, in emerging economies, these entities are scarcely inclined towards implementation of newer environmental policies as they're oblivious to perils of excessive carbon emissions and its consequences. Therefore, it is essential to look for sustainability measures to secure a system that reduces the carbon footprint and ultimately reaches a zero-carbon future. This research proposes a method to identify and assess the barriers of carbon regulatory policies (CRPs) so that advancements in carbon emission reduction practices can be pursued; we present a specific focus on developing nations. An integrated multi-criteria decision-making approach is proposed to achieve environmental sustainability. Initially, the Best Worst Method is used to determine the relative importance of the barriers in the implementation of regulatory policies. Subsequently, we utilize Decision Making Trial and Evaluation Laboratory to establish interrelationships among the barriers of carbon policies. To elucidate the application of the proposed novel framework, a case considering multiple manufacturing firms with multiple stakeholders in India is examined.

Characterization of mosaic diamond wafers and hot-filament epilayers by using HR-EBSD technics

Tungsten (W) incorporated diamond film was homoepitaxially grown on a mosaic diamond wafer by hot-filament chemical vapor deposition. The crystallinity of the W incorporated epilayer and the original mosaic diamond wafer was characterized by using high-resolution electron backscatter diffraction (HR-EBSD). From kernel average misorientation maps (KAM) derived from HR-EBSD measurements, it was found that the numerical value of KAM and the width of coalescence boundary region (CBR) of W incorporated epilayers were much smaller than those of original mosaic diamond wafers and not only in the CBR but also in the bulk area, the crystalline quality of the W incorporated epilayer is better than that of the original mosaic diamond wafers. It is suggested that the primal role for the W incorporation is strain reduction near threading dislocations during growth.

The Meiosis-Specific Crs1 Cyclin Is Required for Efficient S-Phase Progression and Stable Nuclear Architecture.

Cyclins and CDKs (Cyclin Dependent Kinases) are key players in the biology of eukaryotic cells, representing hubs for the orchestration of physiological conditions with cell cycle progression. Furthermore, as in the case of meiosis, cyclins and CDKs have acquired novel functions unrelated to this primal role in driving the division cycle. Meiosis is a specialized developmental program that ensures proper propagation of the genetic information to the next generation by the production of gametes with accurate chromosome content, and meiosis-specific cyclins are widespread in evolution. We have explored the diversification of CDK functions studying the meiosis-specific Crs1 cyclin in fission yeast. In addition to the reported role in DSB (Double Strand Break) formation, this cyclin is required for meiotic S-phase progression, a canonical role, and to maintain the architecture of the meiotic chromosomes. Crs1 localizes at the SPB (Spindle Pole Body) and is required to stabilize the cluster of telomeres at this location (bouquet configuration), as well as for normal SPB motion. In addition, Crs1 exhibits CDK(Cdc2)-dependent kinase activity in a biphasic manner during meiosis, in contrast to a single wave of protein expression, suggesting a post-translational control of its activity. Thus, Crs1 displays multiple functions, acting both in cell cycle progression and in several key meiosis-specific events.

A primal role for the vestibular sense in the development of coordinated locomotion.

Mature locomotion requires that animal nervous systems coordinate distinct groups of muscles. The pressures that guide the development of coordination are not well understood. To understand how and why coordination might emerge, we measured the kinematics of spontaneous vertical locomotion across early development in zebrafish (Danio rerio) . We found that zebrafish used their pectoral fins and bodies synergistically during upwards swims. As larvae developed, they changed the way they coordinated fin and body movements, allowing them to climb with increasingly stable postures. This fin-body synergy was absent in vestibular mutants, suggesting sensed imbalance promotes coordinated movements. Similarly, synergies were systematically altered following cerebellar lesions, identifying a neural substrate regulating fin-body coordination. Together these findings link the vestibular sense to the maturation of coordinated locomotion. Developing zebrafish improve postural stability by changing fin-body coordination. We therefore propose that the development of coordinated locomotion is regulated by vestibular sensation.

106 Randel Lecture: The history of the discovery of the Steroidogenic Acute Regulatory (StAR) Protein

Abstract This two-part presentation regarding acute regulation of steroid biosynthesis documents discovery of the StAR protein and resolves controversy regarding mitochondrial cholesterol transport. The acute regulation of steroid biosynthesis was known to require de novo synthesis of a regulator protein to mediate the transfer of cholesterol, the substrate for steroids, from the outer to the inner mitochondrial membrane where it was converted to pregnenolone by the cytochrome P450 side chain cleavage enzyme. We discovered a novel protein that was tightly correlated with steroid biosynthesis and had the requisite characteristics for the putative acute regulator of cholesterol transfer for steroid synthesis. Further studies confirmed that StAR protein is an indispensable component in the process of mitochondrial uptake of the cholesterol substrate for steroidogenesis. The translocator protein (TSPO) is a mitochondrial outer membrane protein suggested to import cholesterol to the inner mitochondrial membrane. However, it was demonstrated in vivo in Leydig cell specific TSPO conditional knockout mice that TSPO was not required for testosterone production or fertility. Similarly, global TSPO knockout (TSPO/-) mice were viable and fertile with fecundity equivalent to TSPO floxed (TSPOfl/fl) controls. Adrenal and gonadal steroidogenesis did not differ between TSPOfl/ fl and TSPO-/- mice. In vitro use of different steroidogenic cell line models (MA-10, MLTC, Y-1, H295R and R2C) demonstrated that siRNA-knockdown of TSPO did not affect steroidogenesis. Also, CRISPR/ Cas9-mediated TSPO deletion did not affect MA-10 cell steroidogenesis. These results directly 1) refute the suggestion that TSPO is indispensable for viability and steroid hormone biosynthesis; and, 2) substantiate the primal role of the StAR protein as the rate limiting factor in steroid hormone biosynthesis.

Creditor Information, Creditor Rights, and Corporate Bonds

This paper appraises the relative impact of creditor information and creditor rights on corporate bond issues. I employ a large panel of 3,969 bond issuers from 54 countries, between 2013 and 2017. I constantly find that an increase in creditor information is associated with an increase in the amounts of bonds issued. On the opposite, I document a very limited impact of creditor protection. Considering both simultaneously, only the positive impact of information persists, supporting a primal role of information on bond markets. I show that the positive effect of information originates from both the supply- and demand-side of the bond market, with lower coupon rates, less restrictive covenants, longer maturities and more frequent issues. I further document how creditor information alleviates issues stemming from firm’s opacity and firm’s risk. Results are robust to different specifications and robustness tests.

Life cycles of coral-associated-viruses related to diffrent health states collected in Hang Rai - Ninh Thuan, Vietnam

In recent years, coral associated viruses could play primal role in protecting coral health, yet their ecological traits are poorly investigated. In this study, the abundance and life cycles (lysogeny and lytic) of viruses associated with contrasted coral health states (healthy and bleached) were studied and compared. In addition, the density of bacteria and Symbiodium were also be determined by epifluorescence microscopy. The results showed that viral abundance increased 1.2 times and bacterial abundance decreased 0.6 times in healthy corals compared to bleached ones. The number of Symbiodinium significantly reduced 5.6 times in bleached corals compared to healthy corals. One of the most important results in this study was the coexistence of the both lysogenic and lytic viruses in healthy coral. The fraction of lysogenic bacteria was 2 times higher in healthy corals than in bleached ones. Healthy corals carried 3.5-fold more viral production than the diseased ones. Overall, the results showed that, in healthy state, corals might be a favorable environment for viral infection, promoting the development of both temperate and virulent phages. This coexistence might be crucial for coral viability through the complex links with their coral-associated bacterial hosts.

Energy Based Pharmacophore Modelling and Docking Studies for Determining Potent Inhibitor Against M2 Proton Channel of Influenza Virus

Abstract: M2 protein, a crucial glycoprotein present on the viral envelope of Influenza A virus plays an important role in the replication and budding of influenza A virus in the host organism. Due to its primal role in the life cycle of influenza A virus, it is targeted by many potent drugs like amantadine and rimantadine. The emergence of M2 protein mutants in the recent years has rendered these drugs ineffective. Keeping this in our minds, an investigation was performed to determine potent inhibitors of M2 protein using computational strategies like e-pharmacophore based virtual screening and molecular docking. A three dimensional pharmacophore model was generated based on the chemical features using PHASE module of Schrödinger Suite. Subsequently, molecules with same pharmacophoric features were screened from a total of 8621 molecules available in the DrugBank database using the generated pharmacophore hypothesis. This was followed by molecular docking of the screened molecules using three methodologies namely HTVS, SP and XP. Ligand filtration was also done to obtain an efficient collection of hit molecules by analysing pharmacokinetic properties by using Qikprop module. Consequently, our study ascertained nine molecules namely, DB00374, DB00770, DB05015, DB08868, DB00631, DB00983, DB01262, DB00837 and DB01048 which were found to possess anti-viral properties. It is also worth mentioning that antiviral activity of these compounds has been previously reported in recent literature. Key words: Influenza A virus, M2 protein, E-Pharmacophore model, Virtual screening, DrugBank database.

Mycobacterium fortuitum-induced ER-Mitochondrial calcium dynamics promotes calpain/caspase-12/caspase-9 mediated apoptosis in fish macrophages

Mycobacterium fortuitum is a natural fish pathogen. It induces apoptosis in headkidney macrophages (HKM) of catfish, Clarias sp though the mechanism remains largely unknown. We observed M. fortuitum triggers calcium (Ca2+) insult in the sub-cellular compartments which elicits pro-apototic ER-stress factor CHOP. Alleviating ER-stress inhibited CHOP and attenuated HKM apoptosis implicating ER-stress in the pathogenesis of M. fortuitum. ER-stress promoted calpain activation and silencing the protease inhibited caspase-12 activation. The study documents the primal role of calpain/caspase-12 axis on caspase-9 activation in M. fortuitum-pathogenesis. Mobilization of Ca2+ from ER to mitochondria led to increased mitochondrial Ca2+ (Ca2+)m load,, mitochondrial permeability transition (MPT) pore opening, altered mitochondrial membrane potential (ΔΨm) and cytochrome c release eventually activating the caspase-9/-3 cascade. Ultra-structural studies revealed close apposition of ER and mitochondria and pre-treatment with (Ca2+)m-uniporter (MUP) blocker ruthenium red, reduced Ca2+ overload suggesting (Ca2+)m fluxes are MUP-driven and the ER-mitochondria tethering orchestrates the process. This is the first report implicating role of sub-cellular Ca2+ in the pathogenesis of M. fortuitum. We summarize, the dynamics of Ca2+ in sub-cellular compartments incites ER-stress and mitochondrial dysfunction, leading to activation of pro-apoptotic calpain/caspase-12/caspase-9 axis in M. fortuitum-infected HKM.

High hepatic exposure of furanocoumarins in Radix Angelica dahuricae is associated with transporter mediated active uptake

Ethnopharmacological relevanceRadix Angelica dahuricae (RAD), the roots of Angelica dahurica (Hoffm.) Benth. & Hook.f. ex Franch. & Sav, is a well-known traditional Chinese medicine (TCM) and has been used for centuries to treat headaches, toothaches, nose congestion, abscesses, furunculoses, and acne. This herb is also one of frequently reported TCMs showing the herb-drug interaction potential. Furanocoumarins are main bioactive components of RAD. Aim of the studyThis study is designed to characterize the tissue distribution profiles of furanocoumarins after oral administration of RAD extract in rats and to explore the mechanism underlying the high hepatic exposure of the major furanocoumarins. Materials and methodsThe tissue distribution of nine furanocoumarins was determined in rats after an oral dose of 0.46g/kg RAD extract using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Unbound fractions (ƒu) of major furanocoumarins, including imperatorin (IM), isoimperatorin (IIM), bergapten (BER) and oxypeucedanin hydrate (OXYH), were measured in rat plasma and selected tissue homogenates (liver, kidney, lung and brain) with Rapid Equilibrium Dialysis (RED) method. The temperature dependent hepatic uptake of IM, IIM, BER and OXYH were evaluated in suspended rat primary hepatocytes at 4°C or 37°C by the oil-spin method. The uptake kinetics was conducted in the cells over a wide concentration range. The furanocoumarins were co-incubated with a panel of transporter inhibitors to investigate the involvement of uptake transporters in the hepatic uptake. The transcellular transport characteristics of IM, IIM, BER and OXYH were further assessed using Caco-2 cell monolayer model. ResultsIM, IIM, BER and OXYH were found to be the major bioactive furanocoumarins in rat plasma and tissues, representing more than 90% exposure for all the detected furanocoumarins. The most concentrative organ of major furanocoumarins was the liver, with liver-to-plasma exposure ratio (Kp,AUC) of 5.1, 6.5 and 4.7 for IM, IIM and BER, and 2.3 for OXYH, respectively. IM, IIM and BER also showed higher concentrations in the kidney with Kp above 2.2. The higher protein binding of the furanocoumarins partially contributed to their higher tissue exposure. In suspended rat primary hepatocyte, the hepatic uptake of IM, IIM, BER and OXYH was temperature-dependent, with considerably higher uptake at 37°C than at 4°C. Uptake kinetics indicated that the hepatic uptake of IM, IIM, BER and OXYH involved both active transport and passive diffusion processes. For IM, IIM and BER, the contribution of the active transport was greater than the passive process, with the CLactive/CLuptake > 72%. Ritonavir (RTN) and cyclosporine A (CsA), the known inhibitors of organic anion transporting polypeptide (Oatp) significantly inhibited the hepatic uptake of IM and BER, while the inhibitor of the organic anion transporters (Oat) probenecid (PBC) remarkably reduced IIM uptake. In the Caco-2 cell model, the furanocoumarins were highly permeable in the apical to basolateral direction without notable active efflux. ConclusionThe furanocoumarins rapidly and widely distributed into various tissues after oral dose of the RAD extract. IM, IIM, BER and OXYH were the major components detected in both plasma and tissues. Liver was the most distributed tissue of the total and free furanocoumarins. Non-specific protein binding contributed partially to the higher tissue exposures of these bioactive components. The Oatp and Oat mediated active uptake played the primal role in the high hepatic exposure of the furanocoumarins.

Abdul Halim Hasan and his Contributions in Quranic Exegesis in the Malay World

The main objectives of this paper are to expound the exegetical methodologies and contributions of Syeikh Abdul Halim Hasan (d. 1969) in enriching the corpus of Quranic exegetical works in the Malay World. In particular, it focuses on his two main exegetical works entitled Tafsir al-Quran al-Karim which he authored with the assistance of his pupils from 1937 until his demise in 1969, and a work on legal verses of the Quran entitled Tafsir al-Ahkam. Due to its nature, appropriate qualitative methods are applied in this study comprising mainly historical, library research and content analysis. The findings highlight some of the salient features of the exegetical methods of the author and its primal role in influencing the style of exegetical composition by succeeding authors. These books also reflect the concern of the local scholars on the needs of the Muslims for credible religious works and references in languages accessible to them, which have also had its impact on the development in the field of Quranic exegesis in the region.

Dance and early childhood cognition: The Isadora Effect

ABSTRACTThis article proposes the existence of an Isadora Effect: the propositions that motor development plays a primal role in brain development, and the first understanding of symbolic meaning among young children occurs from an understanding of movement and gesture. Anecdotal evidence for the past few decades has demonstrated that dance education and the early use of symbolic movement greatly enhance cognitive development in young children. The author presents (a) background and historical information about dance in early childhood in the United States; (b) research in which physical, emotional, and social development has served the backbone for rationalizing inclusion of dance in early education; (c) The Isadora Effect in early childhood dance education and its relationship to cognitive development, brain development, and intelligence; (d) the 21st Century Skills in relation to early childhood development and dance education; and (e) early childhood dance education and the National Core Arts Standards. The author makes three recommendations for improving the development of American youth relative to early childhood dance education: increased inclusion of creative movement and dance in early childhood curricula, scientific study by the pediatric and neurological communities to investigate effects of motor development and early symbolic movement on brain development and cognitive growth, and ways dance educators can communicate with policy makers about the kinds of support needed to make the first two recommendations a reality.

Human IgE-independent systemic anaphylaxis

Anaphylaxis is a rapidly developing, life-threatening, generalized or systemic allergic reaction that is classically elicited by antigen crosslinking of antigen-specific IgE bound to the high-affinity IgE receptor FcεRI on mast cells and basophils. This initiates signals that induce cellular degranulation with release and secretion of vasoactive mediators, enzymes, and cytokines. However, IgE-independent mechanisms of anaphylaxis have been clearly demonstrated in experimental animals. These include IgG-dependent anaphylaxis, which involves the triggering of mediator release by IgG/antigen complex crosslinking of FcγRs on macrophages, basophils, and neutrophils; anaphylaxis mediated by binding of the complement-derived peptides C3a and C5a to their receptors on mast cells, basophils, and other myeloid cells; and direct activation of mast cells by drugs that interact with receptors on these cells. Here we review the mechanisms involved in these IgE-independent forms of anaphylaxis and the clinical evidence for their human relevance. We conclude that this evidence supports the existence of all 3 IgE-independent mechanisms as important causes of human disease, although practical and ethical considerations preclude their demonstration to the degree of certainty possible with animal models. Furthermore, we cite evidence that different clinical situations can suggest different mechanisms as having a primal role in anaphylaxis and that IgE-dependent and distinct IgE-independent mechanisms can act together to increase anaphylaxis severity. As specific agents become available that can interfere with mechanisms involved in the different types of anaphylaxis, recognition of specific types of anaphylaxis is likely to become important for optimal prophylaxis and therapy.

The Influence of Absolute and Comparative Risk Perceptions on Cervical Cancer Screening and the Mediating Role of Cancer Worry

This research investigates the interrelationships between cancer risk perceptions (absolute and comparative risk perceptions), cancer worry, and cervical cancer screening. Using a nationally representative survey data set (N = 2,304) from the 2012 Health Information National Trends Survey Circle 1, we found that although neither absolute risk perceptions nor comparative risk perceptions exerted a direct impact on women's compliance with the cervical cancer screening recommendation (i.e., that women ages 21 to 65 obtain Pap smear every 3 years; U.S. Preventive Services Task Force, 2012), both types of risk perceptions had an indirect effect on cervical cancer screening through the mediation of cancer worry. These results suggest a primal role of affect in health decision making. Implications of the findings for cancer risk communication are discussed.

“Can Man Be Free If Woman Be A Slave?” A Study in Feminine Leadership in the Works of Percy Bysshe Shelley & Bole Butake

This article studies the primal role of female characters in the works of two dramatists separated by time and space and yet convergent in their philosophical portrayal of the woman as an agent of socio-political mutations. It posits that although Bole Butake is a Cameroonian contemporary playwright, he shares convergent views with Percy Bysshe Shelley, an English Romantic poet who also wrote lyrical dramas, in the portrayal of the front role played by female characters in bringing about change to their societies. The article concludes that though separated by culture and historical epochs, both Butake and Shelley are visionary and insightful in using women to solve the socio-political predicaments of their times.

Role of VASP Phosphorylation Assay in Monitoring the Antiplatelet Therapy

Abstract Dual antiplatelet treatment with clopidogrel and aspirin represents standard regimen in prevention of thromboembolic events in patients with ischemic heart disease undergoing percutaneous coronary intervention (PCI). One of the greatest pitfalls of clopidogrel treatment is large inter-individual variability in response. Large amount of patients does not respond adequately and therefore are not „protected“ even in spite of receiving the therapy. Poor responders are exposed to three-fold increased risk of myocardial infarction, stent thrombosis and cardiac death. Clopidogrel is an antiplatelet prodrug, whose active metabolite inhibits platelet function by irreversible binding to the (adenosine diphosphate) platelet receptor P2Y12. Receptor P2Y12 plays primal role in ADP-mediated platelet activation, and also in mechanism of action of ADP inhibitors such as clopidogrel, prasugrel etc. Reasons stated above, raised the necessity for implementing reliable laboratory test in order to identify the unprotected patients. In an ideal scenario, such test would serve to adjust the dose and guide the individual tailored treatment. Vasodilator Stimulated Phosphoprotein (VASP) is an intracellular platelet protein which is non phosphorylated at basal state. Since its relation in cascade with P2Y12 receptor, VASP phosphorylation corerlates with inhibition of P2Y12 which is the receptor of prime importance in ADP mediated activation of platelets and as is primary target of ADP inhibitors action. Outcome of the assay is represented as the value of platelet reactivity index (PRI), where PRI values above 50% are considered inadequate response to treatment and signal exposure to increased risk of myocardial infarction, post-PCI stent thrombosis and cardiac death. VASP-P flow cytometric assay is emerging into the spotlight as the promising method, mostly for its specificity for ADP inhibitors, better outlook for standardising results and lesser sample manipulation compared to multiple electrode aggregometry.

Structural implication for the impaired binding of W150A mutant LOX-1 to oxidized low density lipoprotein, OxLDL

Lectin-like oxidized lipoprotein (OxLDL) receptor 1, LOX-1, is the major OxLDL receptor expressed on vascular endothelial cells. We have previously reported the ligand-recognition mode of LOX-1 based on the crystal structure of the ligand binding domain (C-type lectin-like domain, CTLD) and surface plasmon resonance analysis, which suggested that the functional significance of the CTLD dimer (the ‘canonical’ dimer) is to harbor the characteristic “basic spine” on its surface. In this study, we have identified the key inter-domain interactions in retaining the canonical CTLD dimer by X-ray structural analysis of the inactive mutant W150A CTLD. The canonical CTLD dimer forms through tight hydrophobic interactions, in which W150 engages in a lock-and-key manner and represents the main interaction. The loss of the Trp ring by mutation to Ala prevents the formation of the canonical dimer, as elucidated from docking calculations using the crystal structure of W150A CTLD. The results emphasize that the canonically formed CTLD dimer is essential for LOX-1 to bind to OxLDL, which supports our proposed view that the basic spine surface present in the correctly formed dimer plays a primal role in OxLDL recognition. This concept provides insight into the pathogenic pattern recognized by LOX-1 as a member of the pattern recognition receptors.