Photodissociation of heme distal methionine in ferrous cytochrome C revealed by subpicosecond time-resolved resonance Raman spectroscopy

Cytochrome c (cyt c) is an electron-transfer heme protein that also binds nitric oxide (NO). In resting cyt c, two endogenous ligands of the heme iron are histidine-18 (His) and methionine-80 (Met) side chains, and NO binding requires the cleavage of one of the axial bonds. Previous femtosecond transient absorption studies suggested the photolysis of either Fe-His or Fe-Met bonds. We aimed at unequivocally identifying the internal side chain that is photodissociated in ferrous cyt c and at monitoring heme structural dynamics, by means of time-resolved resonance Raman (TR3) spectroscopy with approximately 0.6 ps time resolution. The Fe-His stretching mode at 216 cm-1 has been observed in photoproduct TR3 spectra for the first time for a c-type heme. The same transient mode was observed for a model ferrous cyt c N-fragment (residues 1-56) ligated with two His in the resting state. Our TR3 data reveal that upon ferrous cyt c photoexcitation, (i) distal Met side chain is instantly released, producing a five-coordinated domed heme structure, (ii) proximal His side chain, coupled to the heme, exhibits distortion due to strain exerted by the protein, and (iii) alteration in heme-cysteine coupling takes place along with the relaxation of the protein-induced deformations of the heme macrocycle.     

Comparison of lysis methods and preparation protocols for one- and two-dimensional electrophoresis of Aspergillus oryzae intracellular proteins

Filamentous fungal fermentations are used to produce billions of dollars of biochemical and pharmaceutical products annually, yet are plagued by a number of poorly understood problems that would benefit from proteomic analysis. Unfortunately, few publications are available which describe extraction of filamentous fungal proteins for two-dimensional electrophoresis. The goal here was to develop protocols for extraction of fungal proteins, from both wild-type and a recombinant strain of the industrially important filamentous fungi Aspergillus oryzae, to be used for both one- and two-dimensional electrophoresis (1-DE and 2-DE). Because fungal cell walls are exceptionally resistant to fragmentation, four lysis protocols were tested: (i) boiling in strong alkali solution, (ii) boiling in Sodium dodecyl surfate (SDS), (iii) chemical lysis in Y-PER(R) reagent, and (iv) mechanical lysis via rapid agitation with glass beads in a Mini-BeadBeater(R). For both 1-DE and 2-DE, rapid agitation with glass beads was found to be the most efficient extraction method, yielding both mini- and large-format gels with little streaking or spot tailing, and proteins comprising a broad range of molecular weights and pI values.     

A High Cell‐Bearing Capacity Multibore Hollow Fiber Device for Macroencapsulation of Islets of Langerhans

Macroencapsulation of islets of Langerhans is a promising strategy for transplantation of insulin‐producing cells in the absence of immunosuppression to treat type 1 diabetes. Hollow fiber membranes are of interest there because they offer a large surface‐to‐volume ratio and can potentially be retrieved or refilled. However, current available fibers have limitations in exchange of nutrients, oxygen, and delivery of insulin potentially impacting graft survival. Here, multibore hollow fibers for islets encapsulation are designed and tested. They consist of seven bores and are prepared using nondegradable polymers with high mechanical stability and low cell adhesion properties. Human islets encapsulated there have a glucose induced insulin response (GIIS) similar to nonencapsulated islets. During 7 d of cell culture in vitro, the GIIS increases with graded doses of islets demonstrating the suitability of the microenvironment for islet survival. Moreover, first implantation studies in mice demonstrate device material biocompatibility with minimal tissue responses. Besides, formation of new blood vessels close to the implanted device is observed, an important requirement for maintaining islet viability and fast exchange of glucose and insulin. The results indicate that the developed fibers have high islet bearing capacity and can potentially be applied for a clinically applicable bioartificial pancreas.     

Example of a rigorous solution for the gravitational field of a hollow,rotating cylinder

A class of exact solutions-depending upon two dimensionless parameters-to the gravitational field equations is obtained, where the source of the field is a rotating, hollow cylinder of infinite length. The material properties, although highly idealised, nevertheless are not unrealistic and chosen in such a way as to allow elementary integration. The solutions show how the inertial frame inside the cylinder is dragged by the rotating material, and also how the drag continues inside the material and in the outside region. They are exact examples for the Lense-Thirring effect.     

Synthesis and structural study of 2′‐ and 2′,6′‐positioned methyl‐ and nitro‐substituted 3‐(arylhydrazono)pentane‐2,4‐diones

A systematic series of ortho‐methyl‐ and nitro‐substituted arylhydrazones 2–6 formed by Japp–Klingemann reaction between pentane‐2,4‐dione and the respective aryldiazonium salts have been synthesized and studied by X‐ray crystal structure analysis, with added quantum chemical calculations. The optimized molecular geometries based on DFT calculations, enabling determination of relevant rotational barriers, and the calculated bond and ring critical points, using the method of ‘atoms in molecules’, were found to correspond with the experimental data, involving specific molecular conformations and hydrogen‐bonded ring structure dependent on the ortho‐substitution, thus making possible reliable structural prediction of this compound class. Copyright © 2007 John Wiley & Sons, Ltd.     

Compact groups of positive operators on Banach lattices

In this paper, we study groups of positive operators on Banach lattices. If a certain factorization property holds for the elements of such a group, the group has a homomorphic image in the isometric positive operators which has the same invariant ideals as the original group. If the group is compact in the strong operator topology, it equals a group of isometric positive operators conjugated by a single central lattice automorphism, provided an additional technical assumption is satisfied, for which we have only examples. We obtain a characterization of positive representations of a group with compact image in the strong operator topology, and use this for normalized symmetric Banach sequence spaces to prove an ordered version of the decomposition theorem for unitary representations of compact groups. Applications concerning spaces of continuous functions are also considered.     

Speeding up Exponentiation using an Untrusted Computational Resource

We present protocols for speeding up fixed-base variable-exponent exponentiation and variable-base fixed-exponent exponentiation using an untrusted computational resource. In the fixed-base protocols, the exponent may be blinded. In the variable-base protocols, the base may be blinded. The protocols are described for exponentiation in a cyclic group. We describe how to extend them to exponentiation modulo an integer where the modulus is the product of primes with single multiplicity. The protocols provide a speedup of over the square-and-multiply algorithm, where k is the bitlength of the exponent. One application of the protocols is to speed up exponentiation-based verification in discrete log-based signature and credential schemes. The protocols also allow signature verifiers to dynamically choose, for each message, the amount of work it would like to perform to verify the signature. This results in a work-security tradeoff. We introduce a fifth protocol to perform variable-base variable- exponent exponentiation, which also has this feature. Our model allows the trusted resource to perform computations in its idle time. The protocols facilitate the offloading of work to the offline stage, such that the work the trusted resource performs when it has to do an exponentiation is smaller. Our protocols are unconditionally secure.     

Methane oxidation mechanism on Pt(111): a cluster model DFT study

The electronic energy barriers of surface reactions pertaining to the mechanism of the electrooxidation of methane on Pt (111) were estimated with density functional theory calculations on a 10-atom Pt cluster, using both the B3LYP and PW91 functionals. Optimizations of initial and transition states were performed for elementary steps that involve the conversion of CH(4) to adsorbed CO at the Pt/vacuum interface. As a first approximation we do not include electrolyte effects in our model. The reactions include the dissociative chemisorption of CH(4) on Pt, dehydrogenation reactions of adsorbed intermediates (*CH(x) --> *CH(x-1) + *H and *CH(x)O --> *CH(x-1)O + *H), and oxygenation reactions of adsorbed CH(x) species (*CH(x) + *OH --> *CH(x)OH). Many pathways were investigated and it was found that the main reaction pathway is CH(4) --> *CH(3) --> *CH(2) --> *CH --> *CHOH --> *CHO --> *CO. Frequency analysis and transition-state theory were employed to show that the methane chemisorption elementary step is rate-limiting in the above mechanism. This conclusion is in agreement with published experimental electrochemical studies of methane oxidation on platinum catalysts that have shown the absence of an organic adlayer at electrode potentials that allow the oxidation of adsorbed CO. The mechanism of the electrooxidation of methane on Pt is discussed.     

Asymptotic Solutions for Australian Options with Low Volatility

Abstract

In this paper we derive asymptotic expansions for Australian options in the case of low volatility using the method of matched asymptotics. The expansion is performed on a volatility scaled parameter. We obtain a solution that is of up to the third order. In case that there is no drift in the underlying, the solution provided is in closed form, for a non-zero drift, all except one of the components of the solutions are in closed form. Additionally, we show that in some non-zero drift cases, the solution can be further simplified and in fact written in closed form as well. Numerical experiments show that the asymptotic solutions derived here are quite accurate for low volatility.