Ixodes ricinus Salivary Serpin Iripin-8 Inhibits the Intrinsic Pathway of Coagulation and Complement

Tick saliva is a rich source of antihemostatic, anti-inflammatory, and immunomodulatory molecules that actively help the tick to finish its blood meal. Moreover, these molecules facilitate the transmission of tick-borne pathogens. Here we present the functional and structural characterization of Iripin-8, a salivary serpin from the tick Ixodes ricinus, a European vector of tick-borne encephalitis and Lyme disease. Iripin-8 displayed blood-meal-induced mRNA expression that peaked in nymphs and the salivary glands of adult females. Iripin-8 inhibited multiple proteases involved in blood coagulation and blocked the intrinsic and common pathways of the coagulation cascade in vitro. Moreover, Iripin-8 inhibited erythrocyte lysis by complement, and Iripin-8 knockdown by RNA interference in tick nymphs delayed the feeding time. Finally, we resolved the crystal structure of Iripin-8 at 1.89 Å resolution to reveal an unusually long and rigid reactive center loop that is conserved in several tick species. The P1 Arg residue is held in place distant from the serpin body by a conserved poly-Pro element on the P′ side. Several PEG molecules bind to Iripin-8, including one in a deep cavity, perhaps indicating the presence of a small-molecule binding site. This is the first crystal structure of a tick serpin in the native state, and Iripin-8 is a tick serpin with a conserved reactive center loop that possesses antihemostatic activity that may mediate interference with host innate immunity.     

Redox mediation and photomechanical oscillations involving photosensitive cyclometalated Ru(II) complexes, glucose oxidase, and peroxidase

Intact photosensitive cyclometalated RuII derivatives of 2-phenylpyridine or N,N-dimethylbenzylamine cis-[Ru-(C approximately N)(LL)X2]PF6 [C approximately N = o-C6H4-py or o-C6H4CH2NMe2; LL = 1,10-phenanththroline (phen), 2,2'-bipyridine (bpy), or 4,4'-Me2-2,2'-bipyridine (Me2bpy); X = MeCN or pyridine (py)] are efficient mediators of glucose oxidase (GO) from Aspergillus niger and horseradish peroxidase (HRP). Their redox potentials in an aqueous buffer are in the range 0.15-0.35 V versus SCE, and the rate constants for the oxidation GO(red) (where red indicates reduced) by the electrochemically generated RuIII species equal (1.7-2.5) x 10(6) M(-1) s(-1) at pH 7 and 25 degrees C. The redox potentials of all complexes decrease cathodically by 0.4-0.6 V upon irradiation by visible light because of the photoinduced solvolysis of acetonitrile or py ligands. These in situ generated species display an even better mediating performance with HRP, although their behavior toward GO is different. The loading of a ruthenium unit into the protein interior brings about large catalytic currents in a self-assembled system GO-Ru-D-glucose. The estimated rate constant for intramolecular electron transfer from FADH2 of the active site at RuIII, k(intra), equals 4.4 x 10(3) s(-1). This suggests that the distance between the redox partners is around 19 A. The value of 21 A was obtained through the docking analysis of a possible closest-to-FAD localization of a Ru-containing fragment derived from the irradiated complex cis-[Ru(o-C6H4-py)-(phen)(MeCN)2]PF6. The operational stability of the GO-Ru assemblies depends on the nature of complex used, the highest being observed for cis-[Ru(o-C6H4-py)(Me2-bpy)(MeCN)2]PF6 (2). UV-vis studies of interaction of 2 with GO revealed photomechanical oscillations in the system GO-Ru-D-glucose. When irradiated complex 2 is mixed with GO and D-glucose, the absorbance at 510 nm increases because of the enzymatic reduction of RuIII to RuII. The absorbance drops rapidly and then increases as in the first cycle after shaking the reaction solution. Many cycles are possible, and the rate of absorbance increase does not depend on a cycle number. A plausible mechanism of the oscillations is presented.     

"Attenuation of Gastric Lesions by Psychological Aspects of Aggression in Rats": Erratum.

Reports an error in the original article by Jay M. Weiss, Larissa A. Pohorecky, Sherry Salman, and Michael Gruenthal (Journal of Comparative & Physiological Psychology, 1976[Mar], Vol 90[3], 252-259). The figures on pages 255 and 257 should be reversed as Figure 4 now bears the caption of Figure 3 and vice versa. The figures with their correct captions are provided. (The following abstract of this article originally appeared in record 1976-20292-001.) In a study with 80 male albino rats, Ss that fought with each other in response to electric shock showed reduced gastric lesions in comparison with Ss that received the same shocks alone so that fighting behavior did not occur. Also, gastric lesions were similarly reduced in Ss that fought even though they could not physically contact one another because of a barrier between them. In this case, the "protective" effect of fighting derived from the release or display of fighting behavior and did not require physical combat. A 2nd experiment with 48 rats showed that Ss that received shock together but did not engage in fighting behavior showed no reduction of gastric lesions, so that the protective effect of fighting was not an artifact of Ss receiving shock together. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Thermal and microwave assisted polymerization of vinyl acetate catalyzed by cyclometalated ruthenium (II) complexes

Polymerization of vinyl acetate (VAc) was carried out with four different cyclometalated ruthenium(II) catalysts, using CCl₄ as initiator under conventional heating and microwave irradiation. The thermo-polymerizations were solvent-dependent and proceeded to limited conversions without control. Only the polymerization in DMSO mediated by [Ru(η⁶-C₆H₆)(C₆H₄-2-CH₂NMe₂)(MeCN)]PF₆ (1) showed dependence of the molecular weights on conversion. Besides, this complex was the only one active under microwave irradiation. The microwave assisted polymerization was much faster than that under conventional heating, maintaining an increase of the molecular weights with conversion. The chain extensions were quite successful under both conventional heating and microwave irradiation, confirming the participation of ATRP mechanism. The complex 1 was active in atom transfer radical addition of CCl₃Br to VAc substrate providing the adduct in high yield. Mechanistic studies demonstrated the importance of the presence of a labile benzene ligand to increase the catalytic activity of the ruthenium complexes.     

Electromagnetic phase differences in the coherent backscattering enhancement mechanism for random media consisting of large nontransparent spheres

Phase curves of intensity are calculated for light scattering in media randomly packed with large nontransparent spheres (x=125), the surfaces of which reflect according to the Fresnel equations. We consider three values of refractive index: m = 0.73 + i5.93 (metal Al), 1.6 + i1.72 (metal Fe), and 1.5 + i0.1 (black glass). We use a Monte Carlo ray-tracing approach. Different kinds of electromagnetic phase differences of reciprocal trajectories are investigated for the second and third orders of scattering; the highest orders give comparatively small contributions due to the backward-scattering indicatrix of large nontransparent spheres. We find that the main electromagnetic phase difference between the direct and time-reversal (reciprocal) trajectories is the outer phase difference that depends only on the relative positions of the first and last points of the ray reflections and the phase angle. The inner phase difference is connected with the changing path length of the ray inside the medium. This depends on the particle size and the phase angle that is the angle between the source and receiver from the scatterer, i.e., 180 degrees minus the scattering angle. The inner phase difference can give oscillations in the phase curve consisting of second-order components if the medium consists of strictly monodisperse spheres. Usually the coherent backscattering enhancement is calculated ignoring the shadow-hiding effect. We show that accounting for the shadowing of the reciprocal trajectory is important for the formation of the backscattering effect. The third-order scattering surge is a superposition of wide and narrow opposition spikes that correspond to two different types of scattering trajectories, closed and opened ones. The first type is due to scattering by two particles; the second one corresponds to scattering by three particles.     

Resolution function in deep inelastic neutron scattering using the Foil Cycling Technique

New perspectives for epithermal neutron spectroscopy are being opened up by the development of the Resonance Detector (RD) and its use on inverse geometry time of flight (TOF) spectrometers at spallation sources. The most recent result is the Foil Cycling Technique (FCT), which has been developed and applied on the VESUVIO spectrometer operating in the RD configuration. This technique has demonstrated its capability to improve the resolution function of the spectrometer and to provide an effective neutron and gamma background subtraction method. This paper reports a detailed analysis of the line shape of the resolution function in Deep Inelastic Neutron Scattering (DINS) measurements on VESUVIO spectrometer, operating in the RD configuration and employing the FCT. The aim is to provide an analytical approximation for the analyzer energy transfer function, an useful tool for data analysis on VESUVIO. Simulated and experimental results of DINS measurements on a lead sample are compared. The line shape analysis shows that the most reliable analytical approximation of the energy transfer function is a sum of a Gaussian and a power of a Lorentzian. A comparison with the Double Difference Method (DDM) is also discussed. It is shown that the energy resolution improvement for the FCT and the DDM is almost the same, while the counting efficiency is a factor of about 1.4 higher for the FCT.