Plasma Contact Research Articles

Characterization of plasma contacts to cadmium telluride photodetector in ultrathin gas discharge cell

In the present work, the physical properties at the contact of single crystal cadmium telluride semiconductor with gas discharge plasma have been investigated. It is shown that charge carriers in the plasma, together with incident infrared radiation, contribute to the enhancement of the photocurrent in the gas discharge cell. At sufficiently high voltages (more than 2.5 kV) in the gas discharge cell, positive feedback associated with the effect of the plasma on the semiconductor surface is observed. The results of theoretical calculations and experimental experience are in satisfactory agreement, from which the physical meaning of the proportionality coefficient is determined, taking into account the plasma effect on the photoconductivity of the photodetector. The use of a double plasma contact contributes to the damping of the spatial instabilities of the photocurrents in the gas discharge cell, allowing the use of low-resistance photodetectors at the input of the device. Similar results at room temperature have been obtained for the first time on the basis of single-crystal cadmium telluride.

Development and validation of an in vitro model to study thrombin generation on the surface of catheters in platelet-poor and platelet-rich plasma

IntroductionCoagulation activation on medical devices remains a significant problem as it can lead to dramatic thromboembolic complications. Understanding its poorly described mechanisms and finding optimal pharmacological prevention means is crucial to improve patient safety. MethodsWe developed an in vitro model to study thrombin generation (TG) initiated by the contact of plasma with the surface of catheters. Interventional cardiology catheters were cut into segments and inserted in the bottom of multi-well plates; TG was then measured with the calibrated automated thrombogram (CAT). Model performance (analytical, intra- and inter-individual variability) was investigated and compared with activation of thrombin generation by tissue factor (TF) or contact pathway activator (ellagic acid), in the presence (PRP) and absence (PPP) of platelets. Model response to unfractionated heparin (UFH) was also assessed. ResultsTG was greater when measured in presence of catheter segments, compared to conditions without activators. The analytical variability of the model was good (CV ≤ 5 %), both with PPP and PRP. Intra-individual variability was between 15 and 30 % with PPP and between 10 and 15 % with PRP. Inter-individual variability was between 15 and 30 % with both kinds of plasma samples. The analytical performance of the catheter-initiated TG model was equivalent to that observed when TG was initiated with TF or ellagic acid. Catheter-initiated TG was measurable until 0.1 IU/mL UFH with PPP and until 1.0 IU/mL UFH with PRP, highlighting the crucial requirement of platelets. ConclusionOur model is suitable for studying TG initiated with catheters. Inhibition of TG by UFH is overestimated in the absence of platelets.

Study of plasma activated water effect on heavy metal bioaccumulation by Cannabis sativa Using Laser-Induced Breakdown Spectroscopy

Contamination of the environment with toxic metals such as cadmium or lead is a worldwide issue. The accumulator of metals Cannabis sativa L. has potential to be utilized in phytoremediation, which is an environmentally friendly way of soil decontamination. Novel non-thermal plasma-based technologies may be a helpful tool in this process. Plasma activated water (PAW), prepared by contact of gaseous plasma with water, contains reactive oxygen and nitrogen species, which enhance the growth of plants. In this study, C. sativa was grown in a short-term toxicity test in a medium which consisted of plasma activated water prepared by dielectric barrier discharge with liquid electrode and different concentrations of cadmium or lead. Application of PAW on heavy metal contaminated C. sativa resulted in increased growth under Pb contamination as was determined by ecotoxicology tests. Furthermore, the PAW influence on the bioaccumulation of these metals as well as the influence on the nutrient composition of plants was studied primarily by applying Laser-induced breakdown spectroscopy (LIBS). The LIBS elemental maps show that C. sativa accumulates heavy metals mainly in the roots. The results present a new proof-of-concept in which PAW could be used to improve the growth of plants in heavy metal contaminated environment, while LIBS can be implemented to study the phytoremediation efficiency.

Structural basis for the inhibition of βFXIIa by garadacimab

Activated FXII (FXIIa) is the principal initiator of the plasma contact system and can activate both procoagulant and proinflammatory pathways. Its activity is important in the pathophysiology of hereditary angioedema (HAE). Here, we describe a high-resolution cryoelectron microscopy (cryo-EM) structure of the beta-chain from FXIIa (βFXIIa) complexed with the Fab fragment of garadacimab. Garadacimab binds to βFXIIa through an unusually long CDR-H3 that inserts into the S1 pocket in a non-canonical way. This structural mechanism is likely the primary contributor to the inhibition of activated FXIIa proteolytic activity in HAE. Garadacimab Fab-βFXIIa structure also reveals critical determinants of high-affinity binding of garadacimab to activated FXIIa. Structural analysis with other bona fide FXIIa inhibitors, such as benzamidine and C1-INH, reveals a surprisingly similar mechanism of βFXIIa inhibition by garadacimab. In summary, the garadacimab Fab-βFXIIa structure provides crucial insights into its mechanism of action and delineates primary and auxiliary paratopes/epitopes.

Development of a Carbon Nanotube-Enhanced FAS Bilayer Amphiphobic Coating for Biological Fluids.

This study reports the development of a novel amphiphobic coating. The coating is a bilayer arrangement, where carbon nanotubes (CNTs) form the underlayer and fluorinated alkyl-silane (FAS) forms the overlayer, resulting in the development of highly amphiphobic coatings suitable for a wide range of substrates. The effectiveness of these coatings is demonstrated through enhanced contact angles for water and artificial blood plasma fluid on glass, stainless steel, and porous PTFE. The coatings were characterized using Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), atomic force microscopy (AFM), and contact angle (CA) measurements. The water contact angles achieved with the bilayer coating were 106 ± 2°, 116 ± 2°, and 141 ± 2° for glass, stainless steel, and PTFE, respectively, confirming the hydrophobic nature of the coating. Additionally, the coating displayed high repellency for blood plasma, exhibiting contact angles of 102 ± 2°, 112 ± 2°, and 134 ± 2° on coated glass, stainless steel, and PTFE surfaces, respectively. The presence of the CNT underlayer improved plasma contact angles by 29%, 21.7%, and 16.5% for the respective surfaces. The presence of the CNT layer improved surface roughness significantly, and the average roughness of the bilayer coating on glass, stainless steel, and PTFE was measured to be 488 nm, 301 nm, and 274 nm, respectively. Mechanistically, the CNT underlayer contributed to the surface roughness, while the FAS layer provided high amphiphobicity. The maximum effect was observed on modified glass, followed by stainless steel and PTFE surfaces. These findings highlight the promising potential of this coating method across diverse applications, particularly in the biomedical industry, where it can help mitigate complications associated with device-fluid interactions.

Abstract 15197: Differential Impact of Factor XII, Factor XI and Prekallikrein Deficiency on Thrombosis: A Pro-Thrombotic Vascular Action of Bradykinin

Objective: Factor XII (FXII) initiates the plasma contact system that triggers the intrinsic coagulation pathway and the kallikrein-kinin pathway via factor XI (FXI) and plasma prekallikrein (PK), respectively. The importance of the contact system for thrombosis sparing hemostasis is well established, however a comprehensive head-to-head comparison of the three proteases in arterial thrombosis is missing. Approach and Results: We produced mice deficient in FXII ( F12 -/- ), FXI ( F11 -/- ) and PK ( Klkb1 -/- ) and compared them in two distinct thrombosis models. While all (100%) of WT mice formed occlusive thrombi upon 30% ferric chloride challenge, carotid artery occlusion was reduced to 0, 4.8, 27.3 % in F12 -/- , F11 -/- and Klkb1 -/- mice (n=14-22), respectively. Upon photochemical-induced cremaster arteriolar thrombosis, occlusion time (s) prolonged to 226.1±60.4, 123.8±32.0 and 87.8±25.3 (mean±SEM, n=10-15) respectively in F12 -/- , F11 -/- and Klkb1 -/- mice, largely exceeding WT levels (38.3±12.1). A discordance was observed between in vivo thrombotic effects of the proteases and their roles for ex vivo coagulation, which ranked FXII≈FXI>PK as assayed by thromboelastography and activated partial thromboplastin time. Activation of FXII promoted thrombosis in WT and Klkb1 -/- mice, and impressively, it also accelerated thrombosis in F11 -/- mice, increasing activated PK activity and plasma bradykinin formation. Infusion of bradykinin and blockade of degradation of endogenous bradykinin increased platelet-endothelium adhesion and promoted thrombotic occlusions, which involved bradykinin mediated endothelial cell-release of platelet activating factor, rather than direct activation of platelets by bradykinin. Conclusion: FXII, FXI and PK differentially mediate experimental thrombosis in vivo and FXII/FXI appear as superior targets for thrombosis as compared to PK. PK- bradykinin axis activated by FXII contributes to thrombosis in vivo .

Scaling Down the Great Egypt Pyramids to Enhance CO2 Splitting in a Micro DBD Reactor

The CO2 splitting reaction has been investigated in a plate-to-plate micro DBD reactor with a high voltage electrode having pyramid charge injection points. The presence of sharp points (pyramids) creates zones with enhanced electric field around them. The minimum discharge voltage in the pyramid micro DBD reactor reduced from 6.5 to 5.2 kV (peak-to-peak). At the same time, the CO2 conversion increased 1.5 times as compared to that in the reactor with a flat electrode. Lowering the discharge gap from 0.50 to 0.25 mm resulted in more intense microdischarges, further increasing CO2 conversion by 1.3 times. At the same time, the energy efficiency increased further by 1.3 times. There exists an optimum residence time of 0.5 ms as a result of an interplay between plasma contact time and flow non-uniformity. The highest energy efficiency of 20% was obtained at a 3 W power, achieving a CO2 conversion of 16%.

Rotating Packed Bed Reactor Integrated with Plasma: Mass Transfer Intensification and Wastewater Treatment

When oxygen plasma contacts with liquid, abundant reactive species are generated to degrade the contaminant. Ozone is the main reactive species in the gas phase, but only part of it is utilized during wastewater treatment owing to its low solubility. In this work, a novel rotating packed bed reactor integrated with plasma (plasma-RPB) was developed to fully utilize ozone and enhance wastewater treatment. The ozone–water mass transfer efficiency of the plasma-RPB with different liquid distributor structures was evaluated. Subsequently, a dimensionless correlation for predicting the volumetric mass transfer coefficient of the plasma-RPB with the rotating cylinder was established, and the deviation between the predicted and experimental values was within ±6%. Employing the plasma-RPB, sulfamethoxazole and TOC removal efficiency reached up to 87.6 and 82.7% within 5 min, respectively, exhibiting prominent degradation performance compared with other advanced oxidation processes. This work provides an effective route for wastewater treatment.

Vascular Dysfunction in Alzheimer's Disease: Alterations in the Plasma Contact and Fibrinolytic Systems.

Alzheimer's disease (AD) is the most common neurodegenerative disease, affecting millions of people worldwide. The classical hallmarks of AD include extracellular beta-amyloid (Aβ) plaques and neurofibrillary tau tangles, although they are often accompanied by various vascular defects. These changes include damage to the vasculature, a decrease in cerebral blood flow, and accumulation of Aβ along vessels, among others. Vascular dysfunction begins early in disease pathogenesis and may contribute to disease progression and cognitive dysfunction. In addition, patients with AD exhibit alterations in the plasma contact system and the fibrinolytic system, two pathways in the blood that regulate clotting and inflammation. Here, we explain the clinical manifestations of vascular deficits in AD. Further, we describe how changes in plasma contact activation and the fibrinolytic system may contribute to vascular dysfunction, inflammation, coagulation, and cognitive impairment in AD. Given this evidence, we propose novel therapies that may, alone or in combination, ameliorate AD progression in patients.

Anti-HK antibody inhibits the plasma contact system by blocking prekallikrein and factor XI activation invivo.

A dysregulated plasma contact system is involved in various pathological conditions, such as hereditary angioedema, Alzheimer disease, and sepsis. We previously showed that the 3E8 anti-high molecular weight kininogen (anti-HK) antibody blocks HK cleavage and bradykinin generation in human plasma exvivo. Here, we show that 3E8 prevented not only HK cleavage but also factor XI (FXI) and prekallikrein (PK) activation by blocking their binding to HK in mouse plasma invivo. 3E8 also inhibited contact system-induced bradykinin generation invivo. Interestingly, FXII activation was also inhibited, likely because of the ability of 3E8 to block the positive feedback activation of FXII by kallikrein (PKa). In human plasma, 3E8 also blocked PK and FXI binding to HK and inhibited both thrombotic (FXI activation) and inflammatory pathways (PK activation and HK cleavage) of the plasma contact system activation exvivo. Moreover, 3E8 blocked PKa binding to HK and dose-dependently inhibited PKa cleavage of HK. Our results reveal a novel strategy to inhibit contact system activation invivo, which may provide an effective method to treat human diseases involving contact system dysregulation.

Effect of Non-Equilibrium Plasma on Microorganisms Colonizing Diatomaceous Earth after the Beer Filtration Process

The present study evaluates the antimicrobial potential of non-equilibrium plasma against microorganisms isolated from diatomaceous earth, which is a waste product from the beer filtration process. For evaluation, waste diatomaceous earth from an industrial brewery was treated with non-equilibrium plasma using a glidearc reactor. The temperature of the treated samples was monitored. The effect of plasma on the morphology of the treated material was investigated microscopically. Plasma can affect the treated material in various ways and change its physicochemical properties. Consequently, the scope of potential plasma applications is constantly expanding from material technologies to decontamination applications in the food industry, environmental protection and medicine and stimulates activities in, for example, agriculture and medicine. At the same time, microbiological analyzes were carried out to determine the presence of selected groups of microorganisms on diatomaceous earth before and after plasma treatment. The study revealed that the porous structure of diatomaceous earth is not an obstacle to the effective removal of microorganisms from it using plasma. A significant decrease in the amount of both bacteria and yeast relative to the control (non-plasma samples) was observed with the increasing contact time of the diatomaceous earth with the plasma. The numbers of bacteria and yeast decreased by a maximum of 2.2 log10 CFU∙g−1 and 1.72 log10 CFU∙g−1 (30 min of plasma contact), respectively. The obtained results are extremely promising and encourage further, in-depth research to optimize the plasma process and its effect on microorganisms.

A site on factor XII required for productive interactions with polyphosphate

BackgroundDuring plasma contact activation, factor XII (FXII) binds to surfaces through its heavy chain and undergoes conversion to the protease FXIIa. FXIIa activates prekallikrein and factor XI (FXI). Recently, we showed that the FXII first epidermal growth factor-1 (EGF1) domain is required for normal activity when polyphosphate is used as a surface. ObjectivesThe aim of this study was to identify amino acids in the FXII EGF1 domain required for polyphosphate-dependent FXII functions. MethodsFXII with alanine substitutions for basic residues in the EGF1 domain were expressed in HEK293 fibroblasts. Wild-type FXII (FXII-WT) and FXII containing the EGF1 domain from the related protein Pro-HGFA (FXII-EGF1) were positive and negative controls. Proteins were tested for their capacity to be activated, and to activate prekallikrein and FXI, with or without polyphosphate, and to replace FXII-WT in plasma clotting assays and a mouse thrombosis model. ResultsFXII and all FXII variants were activated similarly by kallikrein in the absence of polyphosphate. However, FXII with alanine replacing Lys73, Lys74, and Lys76 (FXII-Ala73,74,76) or Lys76, His78, and Lys81 (FXII-Ala76,78,81) were activated poorly in the presence of polyphosphate. Both have <5% of normal FXII activity in silica-triggered plasma clotting assays and have reduced binding affinity for polyphosphate. Activated FXIIa-Ala73,74,76 displayed profound defects in surface-dependent FXI activation in purified and plasma systems. FXIIa-Ala73,74,76 reconstituted FXII-deficient mice poorly in an arterial thrombosis model. ConclusionFXII Lys73, Lys74, Lys76, and Lys81 form a binding site for polyanionic substances such as polyphosphate that is required for surface-dependent FXII function.

Titanium is a potent inducer of contact activation: implications for intravascular devices

BackgroundTitanium (Ti) and its alloys are widely used in manufacturing medical devices because of their strength and resistance to corrosion. Although Ti compounds are considered compatible with blood, they appear to support plasma contact activation and may be thrombogenic. ObjectivesThe objective of this study was to compare Ti and titanium nitride (TiN) with known activators of contact activation (kaolin and silica) in plasma-clotting assays and to assess binding and activation of factor XII, (FXII), factor XI (FXI), prekallikrein, and high-molecular-weight kininogen (HK) with Ti/TiN. MethodsTi-based nanospheres and foils were compared with kaolin, silica, and aluminum in plasma-clotting assays. Binding and activation of FXII, prekallikrein, HK, and FXI to surfaces was assessed with western blots and chromogenic assays. ResultsUsing equivalent surface amounts, Ti and TiN were comparable with kaolin and superior to silica, for inducing coagulation and FXII autoactivation. Similar to many inducers of contact activation, Ti and TiN are negatively charged; however, their effects on FXII are not neutralized by the polycation polybrene. Antibodies to FXII, prekallikrein, or FXI or coating Ti with poly-L-arginine blocked Ti-induced coagulation. An antibody to FXII reduced FXII and PK binding to Ti, kallikrein generation, and HK cleavage. ConclusionTitanium compounds induce contact activation with a potency comparable with that of kaolin. Binding of FXII with Ti shares some features with FXII binding to soluble polyanions but may have unique features. Inhibitors targeting FXII or FXI may be useful in mitigating Ti-induced contact activation in patients with titanium-based implants that are exposed to blood.

Novel concept suppressing plasma heat pulses in a tokamak by fast divertor sweeping

One of the remaining challenges in magnetic thermonuclear fusion is survival of the heat shield protecting the tokamak reactor vessel against excessive plasma heat fluxes. Unmitigated high confinement edge localized mode (ELM) is a regular heat pulse damaging the heat shield. We suggest a novel concept of magnetic sweeping of the plasma contact strike point fast and far enough in order to spread this heat pulse. We demonstrate feasibility of a dedicated copper coil in a resonant circuit, including the induced currents and power electronics. We predict the DEMO ELM properties, simulate heat conduction, 3D particles motion and magnetic fields of the plasma and coil in COMSOL Multiphysics and Matlab. The dominant system parameter is voltage, feasible 18 kV yields 1 kHz sweeping frequency, suppressing the ELM-induced surface temperature rise by a factor of 3. Multiplied by other known mitigation concepts, ELMs might be mitigated enough to ensure safe operation of DEMO.

Computational Assessment on Impurity Sourcing and Transport Using High-Temperature Graphite and Silicon Carbide Plasma-Facing Walls in a Tokamak Environment

Operating with hot tokamak plasma-facing components will be essential in fusion reactors to maximize the thermal efficiency of the blanket. The SOLPS-ITER edge plasma code package and the DIVIMP Monte Carlo impurity tracking code were used in tandem to simulate the effect of active wall heating on impurity sourcing and transport in a DIII-D–size tokamak. The SOLPS-ITER plasma background was generated based on a previous DIII-D discharge and includes the effect of particle drifts. DIVIMP simulations found that actively heating the lower divertor (versus the divertor shelf or the entire wall) was the most efficient way to minimize gross erosion and core impurity influx at temperatures above 1000 K. Replacing the graphite wall with a silicon carbide (SiC) wall yielded a 5 to 20× decrease in the estimated gross erosion rate of carbon, with a maximum decrease observed at a lower divertor temperature of 800 K. Gross erosion of Si from SiC was estimated to be almost 100× lower than that of C from SiC, due primarily to the low impact energy of incident D plasma on the divertor targets. The core impurity influx for SiC walls is predicted to be lower than that with graphite walls, but eroded Si ions appear to migrate preferentially (versus C) to the core due to a more peaked erosion profile closer to the strike points where the ion temperature gradient force drives particles upstream. These predictive simulations suggest that active heating of the plasma-facing wall may both lower wall erosion and improve core performance relative to the “warm” walls of current devices that are typically only heated via plasma contact. Relative reductions in gross erosion and upstream accumulation by using SiC instead of graphite as the wall material strengthen the argument for upgrades to current graphite-clad machines and continued development of SiC first-wall and blanket concepts.

Plasma–induced damage in magnetic tunneling junctions

Plasma–induced damage (PID) in the fabrication of magnetic tunnel junction (MTJ) based magnetic random access memory (MRAM) is studied after the formation of MTJ devices, which excludes process effect directly from the ion beam etching and protection layer deposition. Large degradation in coercivity (Hc) of MTJs is found in comparison with that measured just after the top metal （TM） was formed, which heavily deteriorates MRAM performances. This Hc degradation is found to be mainly introduced in plasma-enhanced chemical vapor deposition (PECVD) for passivation (PA) film after TM formation. The amplitude of the degradation depends on plasma power, but independent of the composition of the PA film. Hc reduction is observed once the plasma is turned on even though no PA deposition is performed. These results suggest that the observed Hc degradation is from PID effect most likely due to the plasma charge damage (PCD) induced in the PECVD process as no direct plasma contact with MTJ devices is involved in the process. It is argued that this PCD effect is due to the charge accumulations at the interface between the free layer and MgO barrier based on electric field–modulated magnetic anisotropy effect. This is further confirmed by an experiment in which Hc degradation is largely cured through partially removing the trapped charges by an annealing process.

Toxicity and virucidal activity of a neon-driven micro plasma jet on eukaryotic cells and a coronavirus.

Plasma medicine is a developing field that utilizes the effects of cold physical plasma on biological substrates for therapeutic purposes. Approved plasma technology is frequently used in clinics to treat chronic wounds and skin infections. One mode of action responsible for beneficial effects in patients is the potent antimicrobial activity of cold plasma systems, which is linked to their unique generation of a plethora of reactive oxygen and nitrogen species (ROS). During the SARS-CoV-2 pandemic, it became increasingly clear that societies need novel ways of passive and active protection from viral airway infections. Plasma technology may be suitable for superficial virus inactivation. Employing an optimized neon-driven micro plasma jet, treatment time-dependent ROS production and cytotoxic effects to different degrees were found in four different human cell lines with respect to their metabolic activity and viability. Using the murine hepatitis virus (MHV), a taxonomic relative of human coronaviruses, plasma exposure drastically reduced the number of infected murine fibroblasts by up to 3000-fold. Direct plasma contact (conductive) with the target maximized ROS production, cytotoxicity, and antiviral activity compared to non-conductive treatment with the remote gas phase only. Strikingly, antioxidant pretreatment reduced but not abrogated conductive plasma exposure effects, pointing to potential non-ROS-related mechanisms of antiviral activity. In summary, an optimized micro plasma jet showed antiviral activity and cytotoxicity in human cells, which was in part ROS-dependent. Further studies using more complex tissue models are needed to identify a safe dose-effect window of antiviral activity at modest toxicity.

Evaluation of hemostasis disorders using the thrombodynamic test in patients with chronic glomerulonephritis with nephrotic syndrome

Nephrotic syndrome (NS) is accompanied by a risk of thrombotic complications due to hypercoagulability. Routine laboratory tests are not sensitive enough to detect these disorders, and therefore the use of integral coagulation tests, including a new thrombodynamic test (TT) in patients with NS, is of high relevance. Using a TT to determine hemostasis disorders in patients with chronic glomerulonephritis (CGN) with NS. The study included 49 patients with CGN, mean age 37 years, of which 25 (51%) women and 24 (49%) men. Of all the examined patients, 20 (40.8%) of people had NS, 29 (59.2%) had no NS. The process of clot formation was assessed by TT. According to TT, 30% (6/20) of patients with NS and 13.7% (4/29) of patients without NS have hypercoagulation with changes in parameters that go beyond the reference values. In patients with NS, an increase in clot density (D), clot formation rate (V) and clot size (CS) was found, especially when albumin decreased below 25 g/l. Negative correlations were found between the levels of albumin, creatinine and clot density (D), which reflects the level of hyperfibrinogenemia, the rate of clot formation (V) and the integral index of coagulation (CS). The results indicate mainly the activation of the plasma hemostasis due to the internal coagulation pathway. However, the correlation of Tlag (delay time for the onset of clot formation after contact of blood plasma with the insert-activator) with serum cholesterol levels may also indicate activation of the extrinsic coagulation pathway. In CGN patients with NS, activation of the plasma hemostasis is noted, as evidenced by an increase in the rate of formation (V) and size of the clot (CS) after 30 minutes, as well as the density of the formed clot (D).

Ayaconin, a novel inhibitor of the plasma contact system from the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis

Transcriptome analysis of the salivary gland cDNA library from a phlebotomine sand fly, Lutzomyia ayacuchensis, identified a transcript coding for the PpSP15/SL1 family protein as the second most abundant salivary component. In the present study, a recombinant protein of the PpSP15/SL1 family protein, designated ayaconin, was expressed in Escherichia coli, and its biological activity was characterized. The recombinant ayaconin purified from the soluble fraction of E. coli lysate efficiently inhibited the intrinsic but not extrinsic blood coagulation pathway. When the target of ayaconin was evaluated using fluorescent substrates of coagulation factors, ayaconin inhibited factor XIIa (FXIIa) activity more efficiently in a dose-dependent manner, suggesting that FXII is the primary target of ayaconin. In addition, incubation of ayaconin with FXII prior to activation effectively inhibited FXIIa activity, whereas such inhibition was not observed when ayaconin was mixed after the production of FXIIa, indicating that ayaconin inhibits the activation process of FXII to produce FXIIa, but not the enzymatic activity of FXIIa. Moreover, ayaconin was shown to bind to FXII, suggesting that the binding of ayaconin to FXII is involved in the inhibitory mechanism against FXII activation. These results suggest that ayaconin plays an important role in the blood-sucking of Lu. ayacuchensis.

Novel salivary antihemostatic activities of long-form D7 proteins from the malaria vector Anopheles gambiae facilitate hematophagy

To successfully feed on blood, hematophagous arthropods must combat the host's natural hemostatic and inflammatory responses. Salivary proteins of blood-feeding insects such as mosquitoes contain compounds that inhibit these common host defenses against blood loss, including vasoconstriction, platelet aggregation, blood clotting, pain, and itching. The D7 proteins are some of the most abundantly expressed proteins in female mosquito salivary glands and have been implicated in inhibiting host hemostatic and inflammatory responses. Anopheles gambiae, the primary vector of malaria, expresses three D7 long-form and five D7 short-form proteins. Previous studies have characterized the AngaD7 short-forms, but the D7 long-form proteins have not yet been characterized in detail. Here, we characterized the A. gambiae D7 long-forms by first determining their binding kinetics to hemostatic agonists such as leukotrienes and serotonin, which are potent activators of vasoconstriction, edema formation, and postcapillary venule leakage, followed by ex vivo functional assays. We found that AngaD7L1 binds leukotriene C4 and thromboxane A2 analog U-46619; AngaD7L2 weakly binds leukotrienes B4 and D4; and AngaD7L3 binds serotonin. Subsequent functional assays confirmed AngaD7L1 inhibits U-46619-induced platelet aggregation and vasoconstriction, and AngaD7L3 inhibits serotonin-induced platelet aggregation and vasoconstriction. It is therefore possible that AngaD7L proteins counteract host hemostasis by scavenging these mediators. Finally, we demonstrate that AngaD7L2 had a dose-dependent anticoagulant effect via the intrinsic coagulation pathway by interacting with factors XII, XIIa, and XI. The uncovering of these interactions in the present study will be essential for comprehensive understanding of the vector-host biochemical interface.