The interaction of the antitoxin DM43 with a snake venom metalloproteinase analyzed by mass spectrometry and surface plasmon resonance

DM43 is a circulating dimeric antitoxin isolated from Didelphis aurita, a South American marsupial naturally immune to snake envenomation. This endogenous inhibitor binds non‐covalently to jararhagin, the main hemorrhagic metalloproteinase from Bothrops jararaca snake venom, and efficiently neutralizes its toxicity. The aim of this study was to apply mass spectrometry (MS) and surface plasmon resonance (SPR) to improve the molecular characterization of this heterocomplex. The stoichiometry of the interaction was confirmed by nanoelectrospray ionization‐quadrupole‐time‐of‐flight MS; from native solution conditions, the complex showed a molecular mass of ~94 kDa, indicating that one molecule of jararhagin (50 kDa) interacts with one monomer of DM43 (43 kDa). Although readily observed in solution, the dimeric structure of the inhibitor was barely preserved in the gas phase. This result suggests that, in contrast to the toxin–antitoxin complex, hydrophobic interactions are the primary driving force for the inhibitor dimerization. For the real‐time interaction analysis, the toxin was captured on a sensor chip derivatized with the anti‐jararhagin monoclonal antibody MAJar 2. The sensorgrams obtained after successive injections of DM43 in a concentration series were globally fitted to a simple bimolecular interaction, yielding the following kinetic rates for the DM43/jararhagin interaction: k_a = 3.54 ± 0.03 × 10⁴ M^?1 s^?1 and k_d = 1.16 ± 0.07 × 10^?5 s^?1, resulting in an equilibrium dissociation constant (K_D) of 0.33 ± 0.06 nM. Taken together, MS and SPR results show that DM43 binds to its target toxin with high affinity and constitute the first accurate quantitative study on the extent of the interaction between a natural inhibitor and a metalloproteinase toxin, with unequivocal implications for the use of this kind of molecule as template for the rational development of novel antivenom therapies. Copyright © 2012 John Wiley & Sons, Ltd.     

Measuring acoustic energy density in microchannel acoustophoresis using a simple and rapid light-intensity method

We present a simple and rapid method for measuring the acoustic energy density in microchannel acoustophoresis based on light-intensity measurements of a suspension of particles. The method relies on the assumption that each particle in the suspension undergoes single-particle acoustophoresis. It is validated by the single-particle tracking method, and we show by proper re-scaling that the re-scaled light intensity plotted versus re-scaled time falls on a universal curve. The method allows for analysis of moderate-resolution images in the concentration range encountered in typical experiments, and it is an attractive alternative to particle tracking and particle image velocimetry for quantifying acoustophoretic performance in microchannels.     

Behavior of captive herring exposed to naval sonar transmissions (1.0-1.6 kHz) throughout a yearly cycle

Atlantic herring, Clupea harengus, is a hearing specialist, and several studies have demonstrated strong responses to man-made noise, for example, from an approaching vessel. To avoid negative impacts from naval sonar operations, a set of studies of reaction patters of herring to low-frequency (1.0-1.5 kHz) naval sonar signals has been undertaken. This paper presents herring reactions to sonar signals and other stimuli when kept in captivity under detailed acoustic and video monitoring. Throughout the experiment, spanning three seasons of a year, the fish did not react significantly to sonar signals from a passing frigate, at received root-mean-square sound-pressure level (SPL) up to 168 dB re 1 μPa. In contrast, the fish did exhibit a significant diving reaction when exposed to other sounds, with a much lower SPL, e.g., from a two-stroke engine. This shows that the experimental setup is sensitive to herring reactions when occurring. The lack of herring reaction to sonar signals is consistent with earlier in situ behavioral studies. The complexity of the behavioral reactions in captivity underline the need for better understanding of the causal relationship between stimuli and reaction patterns of fish.     

Sorption of aldonic acids from ethyl alcohol-water solutions on anion exchangers

Aldonic acids can be retained quantitatively from ethyl alcohol-water solutions by means of anion exchangers in the sulfate form. After the sorption step the aldonic acids can be easily displaced by washing with water. The sorption is explained by a partition mechanism of the same type as demonstrated in earlier work with strongly polar non-electrolytes. Precautions have to be taken to prevent uptake by means of an ion exchange mechanism. The method permits a separation from exchangeable anions and from non-adsorbable solutes.     

Fluorimetric determination of lactate with immobilized lactate dehydrogenase

In measuring inter-phase distribution coefficients (k_{$\text{1}\text{11}$})it is often convenient, and sometimes necessary, to obtain all of the analytical data from one phase. A rigorous formula, applicable to both low and high k_{$\text{1}\text{11}$} values has been derived; measurements of the distribution of mercury, methylmercury and ⁶⁷Cu²⁺ between various pairs of phases with this formula gave k_{$\text{1}\text{11}$} values similar to those obtained from other approaches.     

Microstructural characterization of self-supported 1.6 μm Pd/Ag membranes

The microstructures of self-supported 1.6 μm thick Pd/23 wt% Ag membranes grown by magnetron sputtering and tested for hydrogen permeability under water–gas shift reaction conditions at different temperatures have been studied. The highest hydrogen permeation was observed through the membrane tested at the highest temperature. X-ray diffraction, transmission electron microscopy, scanning electron microscopy and secondary ion mass spectroscopy were used to characterize the membranes. The grain structure is highly textured and the surface topology is roughened as a function of testing temperature. Hydrogen was trapped in the membranes, localized at and near the feed surface, and the level increased with temperature. The Ag composition at some positions on the feed side of the membrane tested at the highest temperature was reduced to around 5 wt%, probably due to the presence of impurities in the feed gas. This dealloying was associated with the presence of Fe impurity and is probably the cause of small satellite peaks in the X-ray diffractograms.     

Structural changes and coordinatively unsaturated metal atoms on dehydration of honeycomb analogous microporous metal-organic frameworks

Porous metal-organic framework compounds with coordinatively unsaturated metal sites on the inner surface of the pores promise to be valuable adsorbents and catalyst systems, either in industrial applications or as model systems to study interactions with guest molecules. The dehydration process of two isostructural microporous coordination polymers, [M2(dhtp)(H2O)2].8 H2O, termed CPO-27-M (M=Co, Zn; H(4)dhtp=2,5-dihydroxyterephthalic acid) was investigated by in situ variable temperature X-ray diffraction. Both compounds contain accessible coordination sites at the metal after complete removal of the solvent. However, despite the analogy of their crystal structures, they behave differently during dehydration. For CPO-27-Co, water desorption is a smooth topotactic process of second order with no concomitant space group change and no increase in microstrain, which is beneficial for the applicability of the material. Removal of the water propagates from the center of the channels outwards. The coordinating water molecule at the metal desorbs only when almost all the bulk water in the pores has disappeared. In contrast, discontinuities in the powder pattern of CPO-27-Zn indicate the occurrence of first-order transitions. The crystal structures of four of the five individual phases could be determined. The structure of the intermediate phase occurring just before the framework is completely evacuated was elusive in respect to full structure solution and refinement, but it is most probably related to the removal of the axis of threefold symmetry. The zinc-based material experiences a significant amount of strain.     

Mixing properties of one-dimensional cellular automata

We study a class of endomorphisms on the space of bi-infinite sequences over a finite set, and show that such a map is onto if and only if it is measure-preserving. A class of dynamical systems arising from these endomorphisms are strongly mixing, and some of them even -mixing. Some of these are isomorphic to the one-sided shift on in both the topological and measure-theoretical sense. Such dynamical systems can be associated to , the Cuntz-algebra of order , in a natural way.

     

Testing the permeability and corrosion resistance of micro-mechanically interlocked joints

Micro-mechanical interlocking (MMI) can be applied to create new and interesting composite materials. We have employed laser structuring to achieve MMI between stainless steel and plastic with extremely high joint strength. However, the water permeability and corrosion resistance of the joint must be examined. For many industrially relevant applications it is important to keep water away from certain parts and to prevent the sample from corroding. A thorough study of the permeability of the interconnected samples at different temperatures and after employing different laser-structuring techniques is conducted. The permeability seems to be consistent with the Hagen–Poiseuille equation independent of the laser structuring technique and is orders of magnitudes larger than the diffusion rate through the plastic. Two different types of corrosion tests have been undertaken, and we show that care must be taken in order not to degrade the corrosion resistance of the sample to an unacceptable level.     

Ultrasonic measurement and characterization of a low concentration system of solid particles in liquid,in high shear flow

The objective of this paper is ultrasonic measurement and characterization of solid particles in liquid (20–40 μm glass beads in water) in high shear flow (1.5–2 m/s). Ultrasonic time dependent signals as well as frequency spectra are analyzed, for simultaneous determination of average particle concentration and average flow speed. As a result, the distribution of sound energy in such concentrated systems at given flow speeds is measured. Influence of flow turbulence is demonstrated in measurements. Also, characteristic behaviors of liquid–particle mixtures like particle clustering and influence of gas bubbles have been investigated. Experimental results are complemented with a discussion of factors that influence measurement uncertainty.