Voltage clamp behavior of iron-nitric acid system as compared with that of nerve membrane

The current-voltage relation for the surface layer of an iron wire immersed in nitric acid was investigated by the voltage clamp technique. Comparing the phase of nitric acid to the axoplasm and the metallic phase to the external fluid medium for the nerve fiber, a striking analogy was found between the voltage clamp behavior of the iron-nitric acid system and that of the nerve membrane. The current voltage curve was found to consist of three parts: (a) a straight line representing the behavior of the resting (passive) membrane, (b) a straight line representing the fully excited (active) state, and (c) an intermediate zone connecting (a) and (b). It was shown that in the intermediate zone, the surface of iron consisted of a fully active patch (or patches) surrounded by a remaining resting area. The phenomenon corresponding to "repetitive firing of responses under voltage clamp" in the nerve membrane was demonstrated in the intermediate zone. The behavior of the cobalt electrode system was also investigated by the same technique. An attempt was made to interpret the phenomenon of initiation and abolition of an active potential on the basis of the thermodynamics of irreversible processes.     

sEMG wavelet-based indices predicts muscle power loss during dynamic contractions

The purpose of this study was to investigate the sensitivity of new surface electromyography (sEMG) indices based on the discrete wavelet transform to estimate acute exercise-induced changes on muscle power output during a dynamic fatiguing protocol. Fifteen trained subjects performed five sets consisting of 10 leg press, with 2 min rest between sets. sEMG was recorded from vastus medialis (VM) muscle. Several surface electromyographic parameters were computed. These were: mean rectified voltage (MRV), median spectral frequency (F_med), Dimitrov spectral index of muscle fatigue (FI_nsm5), as well as five other parameters obtained from the stationary wavelet transform (SWT) as ratios between different scales. The new wavelet indices showed better accuracy to map changes in muscle power output during the fatiguing protocol. Moreover, the new wavelet indices as a single parameter predictor accounted for 46.6% of the performance variance of changes in muscle power and the log-FI_nsm5 and MRV as a two-factor combination predictor accounted for 49.8%. On the other hand, the new wavelet indices proposed, showed the highest robustness in presence of additive white Gaussian noise for different signal to noise ratios (SNRs). The sEMG wavelet indices proposed may be a useful tool to map changes in muscle power output during dynamic high-loading fatiguing task.     

Stability of localized MHD perturbations in confinement systems with closed magnetic field lines

Conditions are determined for the stability of a finite-pressure plasma against perturbations localized near a magnetic field line in a magnetic confinement system without average minimum-B. The marginal stability (ω²=0) is achieved at the pressure profile p∝U ^?5/3 (where $U = \oint {\frac{{dl}}{B}}$ ), provided that the pressure is lower than a certain critical value above which an unstable incompressible mode in which the displacement as a function of the coordinate along the field line has zeros appears at some magnetic field line.     

Stability of production and plant species diversity in managed grasslands: A retrospective study

Plant biodiversity theory suggests that increased plant species diversity contributes to the stability of ecosystems. In managed grasslands, such as pastures, greater stability of herbage production would be beneficial. In this retrospective study, I used data from three reports from the 1930s, 1940s, and 1960s to determine whether increasing mixture complexity contributed to yield stability with time. Yield data from a total of 175 mixtures of grasses and legumes (two to seven species per mixture) in experiments of 3–6 year duration in Connecticut, Pennsylvania, and Utah, USA, were used in the analysis. Regression was used to examine relationships among herbage yield, stability of yield [measured as the interannual coefficient of variation (CV)], and diversity measures [Shannon diversity index (H), species richness (S), and evenness (J)]. In several instances there was no relationship between herbage yield or yield stability and the complexity (number of species) of the mixture. In one experiment, the proportion of legume in the sward seemed to be the controlling factor regarding herbage yield and yield stability. Monocultures and binary mixtures frequently had the highest interannual CV (e.g., less yield stability) than more complex mixtures. Within forage mixtures, however, there was no consistent relationship between herbage yield or yield stability and measures of species diversity. Species identity and composition of forage mixtures may be more important determinants of herbage yield than simply the number of species.     

Fluorescent Protein Voltage Probes Derived from ArcLight that Respond to Membrane Voltage Changes with Fast Kinetics

We previously reported the discovery of a fluorescent protein voltage probe, ArcLight, and its derivatives that exhibit large changes in fluorescence intensity in response to changes of plasma membrane voltage. ArcLight allows the reliable detection of single action potentials and sub-threshold activities in individual neurons and dendrites. The response kinetics of ArcLight (τ1-on ~10 ms, τ2-on ~ 50 ms) are comparable with most published genetically-encoded voltage probes. However, probes using voltage-sensing domains other than that from the Ciona intestinalis voltage sensitive phosphatase exhibit faster kinetics. Here we report new versions of ArcLight, in which the Ciona voltage-sensing domain was replaced with those from chicken, zebrafish, frog, mouse or human. We found that the chicken and zebrafish-based ArcLight exhibit faster kinetics, with a time constant (τ) less than 6ms for a 100 mV depolarization. Although the response amplitude of these two probes (8-9%) is not as large as the Ciona-based ArcLight (~35%), they are better at reporting action potentials from cultured neurons at higher frequency. In contrast, probes based on frog, mouse and human voltage sensing domains were either slower than the Ciona-based ArcLight or had very small signals.     

Ultra‐High Voltage Multijunction Organic Solar Cells for Low‐Power Electronic Applications

Low power electronics are an ideal application for organic photovoltaics (OPV) where a low‐cost OPV device can be integrated directly with a battery to provide a constant power source. We demonstrate ultra‐high voltage small molecule multijunction devices with open circuit voltage (V_OC) values of up to 7V. Optical modelling is employed to aid the optimisation of the complex multi‐layer stacks and ensure current balancing is achieved between sub‐cells, and optimised multijunction devices show power conversion efficiencies of up to 3.4% which is a modest increase over the single junction devices. Sub‐cell donor/acceptor pairs of boron subphthalocyanine chloride (SubPc)/fullerene (C₆₀) and SubPc/Cl₆‐SubPc were selected both for their high V_OC in order to minimise the required number of junctions, but also for their absorption overlap to reduce the spectral dependence of the device performance. As a result, the devices are shown to directly charge a micro‐energy cell type battery under both low illumination intensity white light and monochromatic illumination.     

Cytosolic Selection Systems To Study Protein Stability

Here we describe biosensors that provide readouts for protein stability in the cytosolic compartment of prokaryotes. These biosensors consist of tripartite sandwich fusions that link the in vitro stability or aggregation susceptibility of guest proteins to the in vivo resistance of host cells to the antibiotics kanamycin, spectinomycin, and nourseothricin. These selectable markers confer antibiotic resistance in a wide range of hosts and are easily quantifiable. We show that mutations within guest proteins that affect their stability alter the antibiotic resistances of the cells expressing the biosensors in a manner that is related to the in vitro stabilities of the mutant guest proteins. In addition, we find that polyglutamine tracts of increasing length are associated with an increased tendency to form amyloids in vivo and, in our sandwich fusion system, with decreased resistance to aminoglycoside antibiotics. We demonstrate that our approach allows the in vivo analysis of protein stability in the cytosolic compartment without the need for prior structural and functional knowledge.     

Developing Fast Fluorescent Protein Voltage Sensors by Optimizing FRET Interactions

FRET (Förster Resonance Energy Transfer)-based protein voltage sensors can be useful for monitoring neuronal activity in vivo because the ratio of signals between the donor and acceptor pair reduces common sources of noise such as heart beat artifacts. We improved the performance of FRET based genetically encoded Fluorescent Protein (FP) voltage sensors by optimizing the location of donor and acceptor FPs flanking the voltage sensitive domain of the Ciona intestinalis voltage sensitive phosphatase. First, we created 39 different “Nabi1” constructs by positioning the donor FP, UKG, at 8 different locations downstream of the voltage-sensing domain and the acceptor FP, mKO, at 6 positions upstream. Several of these combinations resulted in large voltage dependent signals and relatively fast kinetics. Nabi1 probes responded with signal size up to 11% ΔF/F for a 100 mV depolarization and fast response time constants both for signal activation (~2 ms) and signal decay (~3 ms). We improved expression in neuronal cells by replacing the mKO and UKG FRET pair with Clover (donor FP) and mRuby2 (acceptor FP) to create Nabi2 probes. Nabi2 probes also had large signals and relatively fast time constants in HEK293 cells. In primary neuronal culture, a Nabi2 probe was able to differentiate individual action potentials at 45 Hz.     

Multi-Virulence-Locus Sequence Typing of Listeria monocytogenes

A multi-virulence-locus sequence typing (MVLST) scheme was developed for subtyping Listeria monocytogenes, and the results obtained using this scheme were compared to those of pulsed-field gel electrophoresis (PFGE) and the published results of other typing methods, including ribotyping (RT) and multilocus sequence typing (MLST). A set of 28 strains (eight different serotypes and three known genetic lineages) of L. monocytogenes was selected from a strain collection (n > 1,000 strains) to represent the genetic diversity of this species. Internal fragments (ca. 418 to 469 bp) of three virulence genes (prfA, inlB, and inlC) and three virulence-associated genes (dal, lisR, and clpP) were sequenced and analyzed. Multiple DNA sequence alignment identified 10 (prfA), 19 (inlB), 13 (dal), 10 (lisR), 17 (inlC), and 16 (clpP) allelic types and a total of 28 unique sequence types. Comparison of MVLST with automated EcoRI-RT and PFGE with ApaI enzymatic digestion showed that MVLST was able to differentiate strains that were indistinguishable by RT (13 ribotypes; discrimination index = 0.921) or PFGE (22 profiles; discrimination index = 0.970). Comparison of MVLST with housekeeping-gene-based MLST analysis showed that MVLST provided higher discriminatory power for serotype 1/2a and 4b strains than MLST. Cluster analysis based on the intragenic sequences of the selected virulence genes indicated a strain phylogeny closely related to serotypes and genetic lineages. In conclusion, MVLST may improve the discriminatory power of MLST and provide a convenient tool for studying the local epidemiology of L. monocytogenes.     

Contribution of genetic factors to noise-induced hearing loss: A human studies review

Noise-induced hearing loss (NIHL) is a complex disease that results from the interaction of genetic and environmental factors. Over the last 10 years there has been a great increase in association studies trying to identify the susceptibility genes for NIHL in humans. They were conducted based on the candidate gene approach and comprised predominantly the group of oxidative stress genes, inner ear potassium recycling pathway genes and monogenic deafness genes, as well as other genes. So far, the most promising results were obtained for two genes encoding potassium ion channels (KCNQ4 and KCNE1), catalase (CAT), protocadherin 15 (PCDH15), myosin 14 (MYH14) and heat shock protein (HSP70), because they were replicated in two (Polish and Swedish) or three (Polish, Swedish and Chinese) populations, and were sufficient in size to yield high power for the detection of a causative allele. Today, the development of high-throughput genotyping methods allows the detection of hundreds and thousands of single nucleotide polymorphisms (SNPs) in a single array which undoubtedly will lead toward identification of new NIHL susceptibility genes. This in turn will contribute to the development of genetics tests that would allow for better protection of noise-exposed individuals and personalized treatment, if necessary.     

Application of TMA-PEG to promote C-phycocyanin extraction from S. platensis in the PEG ATPS

A novel aqueous two phase system (ATPS) using trimethylamine-polyethylene glycols (TMA-PEG) to promote the extraction of C-phycocyanin (C-PC) from S.platensis was introduced. The purity of C-PC (EP) obtained in the ATPS of PEG1000/Na₃PO₄ was increased 2.1 times by the addition of TMA-PEG1000. The purification factor was enhanced from 2.9 to 10.1 when 65% TMA-PEG1000 was added in the system. The ATPS operation must be carried out in the pH range of 6.0-7.0 and at temperatures less than 35 °C for maintaining the stability of C-PC. The partition coefficient and recovery ratio of C-PC increased with the increasing concentration of TMA-PEG. The system parameters like TMA-PEG1000 content, tie line length (TLL), pH, temperature and phase volume ratio (V_r) were screened and optimized using the fractional factorial design and Box-Behnken experiment design. The optimized system is composed of 11.8% PEG1000/TMA-PEG1000 (w/w), 64.42% TMA-PEG1000 (w/w PEG1000) and 9.5% Na₃PO₄ (w/w) with 38.19% TLL (w/w) and 0.89 V_r at pH 6.5 and 25 °C. The obtained value of EP was 5.21 in one-stage ATPS and 6.7 in two-stage ATPS. The recovery ratio of C-PC in the new ATPS extraction system was more than 97%.     

Surveillance of Viruses in Wild Fish Populations in Areas around the Gulf of Cadiz (South Atlantic Iberian Peninsula)

This report describes a viral epidemiological study of wild fish around the Gulf of Cadiz (southwestern Iberian Peninsula) and is focused on infectious pancreatic necrosis virus (IPNV), viral hemorrhagic septicemia virus (VHSV), and viral nervous necrosis virus (VNNV). One fish species (Chelon labrosus) was sampled inside the gulf, at the mouth of the San Pedro River. Another 29 were sampled, in three oceanographic campaigns, at sites around the Bay of Cadiz. The fish were processed individually and subjected to isolation in cell culture and molecular diagnosis. VHSV was not isolated from any species. Thirteen IPNV-type isolates were obtained from barracuda (Sphyraena sphyraena), axillary seabream (Pagellus acarne), common two-banded seabream (Diplodus vulgaris), common pandora (P. erythrinus), Senegal seabream (D. bellottii), and surmullet (Mullus surmuletus). Six VNNV isolates were obtained from axillary seabream, common pandora, black seabream (Spondyliosoma cantharus), red mullet (Mullet barbatus), Lusitanian toadfish (Halobatrachus didactylus), and tub gurnard (Chelidonichtys lucerna). In the river mouth, viruses were detected only after reamplification, obtaining prevalence percentages of IPNV and VNNV (44.4 and 63.0%, respectively) much higher than those observed in the oceanographic campaigns (25.7 and 19.6%, respectively). The opposite results were obtained in the case of VHSV after reamplification: 11.1% in the river mouth and 43.6% in the oceanic locations. Analyzing the results with respect to the proximity of the sampling sites to the coast, an anthropogenic influence on wild fish is suggested and discussed. The type of viruses and the presence of natural reassortants are also discussed.     

Identification of small molecule lead compounds for visceral leishmaniasis using a novel ex vivo splenic explant model system

Background

New drugs are needed to treat visceral leishmaniasis (VL) because the current therapies are toxic, expensive, and parasite resistance may weaken drug efficacy. We established a novel ex vivo splenic explant culture system from hamsters infected with luciferase-transfected Leishmania donovani to screen chemical compounds for anti-leishmanial activity.

Methodology/Principal Findings

This model has advantages over in vitro systems in that it: 1) includes the whole cellular population involved in the host-parasite interaction; 2) is initiated at a stage of infection when the immunosuppressive mechanisms that lead to progressive VL are evident; 3) involves the intracellular form of Leishmania; 4) supports parasite replication that can be easily quantified by detection of parasite-expressed luciferase; 5) is adaptable to a high-throughput screening format; and 6) can be used to identify compounds that have both direct and indirect anti-parasitic activity. The assay showed excellent discrimination between positive (amphotericin B) and negative (vehicle) controls with a Z'' Factor >0.8. A duplicate screen of 4 chemical libraries containing 4,035 compounds identified 202 hits (5.0%) with a Z score of <–1.96 (p<0.05). Eighty-four (2.1%) of the hits were classified as lead compounds based on the in vitro therapeutic index (ratio of the compound concentration causing 50% cytotoxicity in the HepG₂ cell line to the concentration that caused 50% reduction in the parasite load). Sixty-nine (82%) of the lead compounds were previously unknown to have anti-leishmanial activity. The most frequently identified lead compounds were classified as quinoline-containing compounds (14%), alkaloids (10%), aromatics (11%), terpenes (8%), phenothiazines (7%) and furans (5%).

Conclusions/Significance

The ex vivo splenic explant model provides a powerful approach to identify new compounds active against L. donovani within the pathophysiologic environment of the infected spleen. Further in vivo evaluation and chemical optimization of these lead compounds may generate new candidates for preclinical studies of treatment for VL.     

Carbon‐Coated Li3Nd3W2O12: A High Power and Low‐Voltage Insertion Anode with Exceptional Cycleability for Li‐Ion Batteries

The synthesis of carbon‐coated Li₃Nd₃W₂O₁₂ (C‐Li₃Nd₃W₂O₁₂), a low voltage insertion anode (0.3 V vs. Li) for a Li‐ion battery, is reported to exhibit extraordinary performance. The low voltage reversible insertion provides an increase in the energy density of Li‐ion power packs. For instance, C‐Li₃Nd₃W₂O₁₂ delivered an energy density of ≈390 Wh kg^?1 (based on cathode mass loading) when coupled with an LiMn₂O₄ cathode with an operating potential of 3.4 V. Furthermore, excellent cycling profiles are observed for C‐Li₃Nd₃W₂O₁₂ anodes both in half and full‐cell configurations. The full‐cell is capable of delivering very stable cycling profiles at high current rates (e.g., 2 C), which clearly suggests the high power capability of such garnet‐type anodes.     

A FACS-Optimized Screen Identifies Regulators of Genome Stability in Candida albicans

Loss of heterozygosity (LOH) plays important roles in genome dynamics, notably, during tumorigenesis. In the fungal pathogen Candida albicans, LOH contributes to the acquisition of antifungal resistance. In order to investigate the mechanisms that regulate LOH in C. albicans, we have established a novel method combining an artificial heterozygous locus harboring the blue fluorescent protein and green fluorescent protein markers and flow cytometry to detect LOH events at the single-cell level. Using this fluorescence-based method, we have confirmed that elevated temperature, treatment with methyl methanesulfonate, and inactivation of the Mec1 DNA damage checkpoint kinase triggered an increase in the frequency of LOH. Taking advantage of this system, we have searched for C. albicans genes whose overexpression triggered an increase in LOH and identified four candidates, some of which are known regulators of genome dynamics with human homologues contributing to cancer progression. Hence, the approach presented here will allow the implementation of new screens to identify genes that are important for genome stability in C. albicans and more generally in eukaryotic cells.     

Toward Higher Voltage Solid‐State Batteries by Metastability and Kinetic Stability Design

The energy density of battery systems is limited largely by the electrochemical window of the electrolyte. Herein, the combined thermodynamic and kinetic effects of mechanically induced metastability are shown to greatly widen the operational voltage window of solid‐state batteries based on ceramic‐sulfide electrolytes. Solid electrolyte voltage stability up to 10 V is achieved with minimal degradation, far beyond the capability of organic liquid electrolytes. Furthermore, combined experiment, ab initio computation, and theoretical modeling identify the nature of mechanically constrained Li₁₀GeP₂S₁₂ decomposition both within the bulk and at interfaces with cathode materials at very high voltages. Previously unclear kinetic processes are identified that, when properly implemented, can potentially allow solid‐state full cells with remarkably high operational voltages.     

Plant indicator values of a high-phytodiversity country (Italy) and their evidence,exemplified for model areas with climatic gradients in the southern inner Alps

We tested the new Ellenberg–Pignatti indicator values in Festucetalia valesiacae communities of three valley regions (southern inner Alps: Valle d’Aosta, Valtellina, Valle Venosta/Vinschgau). In these landscape types gradients of all value types exist: light (L), temperature (T), moisture (F/M), nitrogen (N), continentality (C) and soil reaction (R). L- and T-values were extended from 9 to 12 in the database of Pignatti due to the higher intensity of these factors in the area in question; in our dataset this is especially relevant for the L-values.In a spatial approach, we compared DCA results with indicator value results (presence–absence) and with climatic data for eight different plant communities. We found comprehensible results in all cases, also for the extension of the L-factor.In a temporal approach, we compared relevés and their indicator values from two time windows: 1930–1950 (Braun-Blanquet) and 1990–1995 (Schwabe and Kratochwil) for six of the eight different plant communities. Both relevé types were made in the same or in similar localities, but no permanent plots existed. Mostly we found in these communities, which are said to have high “stability”, no significant changes on average for any of the indicator values. In the case of N- and R-values there was a trend of increase in the different datasets, and in some cases this trend is significant.In general, the new database has shown the “robustness” of the Ellenberg indicator value model including the extended L-values, and established its usefulness for a synthetic approach in the studied valley systems. In our systems L-, T-, F/M- and C-gradients can be characterised well by means of indicator values and are an excellent tool for explaining DCA results. Caution is necessary especially when correlations of different factors are intermingled in temporal approaches.     

Stability curves of laboratory evolved thermostable mutants of a Bacillus subtilis lipase

Shape of the protein stability curves changes to achieve higher melting temperature. Broadly, these changes have been classified as upward shift (increased ?G_s), rightward shift (increase in T_s) and flattening of the stability curves (decrease in ?C_p). Comparative studies on homologous mesophilic–thermophilic protein pairs highlighted the differential contribution of these three strategies amongst proteins. But unambiguous way of identification of the strategies, which will be preferred for a protein, is still not achieved. We have performed comparative thermodynamic studies using differential scanning calorimeter (DSC) on thermostable variants of a lipase from Bacillus subtilis. These variants are products of 1, 2, 3 and 4 rounds of directed evolution and harbor mutations having definite contribution in thermostability unlike natural thermophilic proteins. We have shown that upward and rightward shift in stability curves are prime strategies in this lipase. Our results along with that from the other study on laboratory evolved xylanase A suggest that optimization of suboptimal thermodynamic parameters is having a dominant influence in selection of thermodynamic strategies for higher thermostability.