Targeted Modifications in Adeno-Associated Virus Serotype 8 Capsid Improves Its Hepatic Gene Transfer Efficiency In Vivo

Recombinant adeno-associated virus vectors based on serotype 8 (AAV8) have shown significant promise for liver-directed gene therapy. However, to overcome the vector dose dependent immunotoxicity seen with AAV8 vectors, it is important to develop better AAV8 vectors that provide enhanced gene expression at significantly low vector doses. Since it is known that AAV vectors during intracellular trafficking are targeted for destruction in the cytoplasm by the host-cellular kinase/ubiquitination/proteasomal machinery, we modified specific serine/threonine kinase or ubiquitination targets on the AAV8 capsid to augment its transduction efficiency. Point mutations at specific serine (S)/threonine (T)/lysine (K) residues were introduced in the AAV8 capsid at the positions equivalent to that of the effective AAV2 mutants, generated successfully earlier. Extensive structure analysis was carried out subsequently to evaluate the structural equivalence between the two serotypes. scAAV8 vectors with the wild-type (WT) and each one of the S/T -> Alanine (A) or K-Arginine (R) mutant capsids were evaluated for their liver transduction efficiency in C57BL/6 mice in vivo. Two of the AAV8-S -> A mutants (S279A and S671A), and a K137R mutant vector, demonstrated significantly higher enhanced green fluorescent protein (EGFP) transcript levels (similar to 9- to 46-fold) in the liver compared to animals that received WT-AAV8 vectors alone. The best performing AAV8 mutant (K137R) vector also had significantly reduced ubiquitination of the viral capsid, reduced activation of markers of innate immune response, and a concomitant two-fold reduction in the levels of neutralizing antibody formation in comparison to WT-AAV8 vectors. Vector bio-distribution studies revealed that the K137R mutant had a significantly higher and preferential transduction of the liver (106 vs. 7.7 vector copies/mouse diploid genome) when compared to WT-AAV8 vectors. To further study the utility of the K137R-AAV8 mutant in therapeutic gene transfer, we delivered human coagulation factor IX (h. FIX) under the control of liver-specific promoters (LP1 or hAAT) into C57BL/6 mice. The circulating levels of h. FIX: Ag were higher in all the K137R-AAV8 treated groups up to 8 weeks post-hepatic gene transfer. These studies demonstrate the feasibility of the use of this novel AAV8 vectors for potential gene therapy of hemophilia B.

Tungsten-based nanomaterials (WO3 & Bi2WO6): Modifications related to charge carrier transfer mechanisms and photocatalytic applications

Heterogeneous photocatalysis is an ideal green energy technology for the purification of wastewater. Although titania dominates as the reference photocatalyst, its wide band gap is a bottleneck for extended utility. Thus, search for non-TiO2 based nanomaterials has become an active area of research in recent years. In this regard, visible light absorbing polycrystalline WO3 (2.4-2.8 eV) and Bi2WO6 (2.8 eV) with versatile structure-electronic properties has gained considerable interest to promote the photocatalytic reactions. These materials are also explored in selective functional group transformation in organic reactions, because of low reduction and oxidation potential of WO3 CB and Bi2WO6 VB, respectively. In this focused review, various strategies such as foreign ion doping, noble metal deposition and heterostructuring with other semiconductors designed for efficient photocatalysis is discussed. These modifications not only extend the optical response to longer wavelengths, but also prolong the life-time of the charge carriers and strengthen the photocatalyst stability. The changes in the surface-bulk properties and the charge carrier transfer dynamics associated with each modification correlating to the high activity are emphasized. The presence of oxidizing agents, surface modification with Cu2+ ions and synthesis of exposed facets to promote the degradation rate is highlighted. In depth study on these nanomaterials is likely to sustain interest in wastewater remediation and envisaged to signify in various green energy applications. (C) 2015 Elsevier B.V. All rights reserved.

Perceptual thresholds for detecting modifications applied to the acoustical properties of a violin

This study is the first step in the psychoacoustic exploration of perceptual differences between the sounds of different violins. A method was used which enabled the same performance to be replayed on different "virtual violins," so that the relationships between acoustical characteristics of violins and perceived qualities could be explored. Recordings of real performances were made using a bridge-mounted force transducer, giving an accurate representation of the signal from the violin string. These were then played through filters corresponding to the admittance curves of different violins. Initially, limits of listener performance in detecting changes in acoustical characteristics were characterized. These consisted of shifts in frequency or increases in amplitude of single modes or frequency bands that have been proposed previously to be significant in the perception of violin sound quality. Thresholds were significantly lower for musically trained than for nontrained subjects but were not significantly affected by the violin used as a baseline. Thresholds for the musicians typically ranged from 3 to 6 dB for amplitude changes and 1.5%-20% for frequency changes. interpretation of the results using excitation patterns showed that thresholds for the best subjects were quite well predicted by a multichannel model based on optimal processing. (c) 2007 Acoustical Society of America.

Contribution à l'étude des modifications hydro-sédimentaires consécutives à la réouverture artificielle de l'embouchure du Comoé à Grand-Bassam

The Bassam artificial inlet was opened (in September 22, 1987) in order to revacuate offshore invasive aquatic plants. This has considerably modified the hydro-sedimentary environment of the lagoonal domain at Grand-Bassam. Tidal currents effect has confered the lagoonal waters an estuarine feature, the saline intrusion (25-35%o) increased beyond the Moossou bridges up to the confluence of the Comoe river. This has led to a cleaning of the river lower course and above all to the bottoms of the lagoon, which were previously confined. The remobilization of mud (from lagoonal bottoms around Bouet Island) evacuated offshore, has exposed lagoonal and oceanic sandy features. This new system remains fragile, with the seasonal (low river-flow) clogging.

Effects of Gear Modifications on the Trawl Performance and Catching Efficiency of the West Coast Upper Continental Slope Groundfish Survey Trawl

Since 1984, annual bottom trawl surveys of the west coast (California–Washington) upper continental slope (WCUCS) have provided information on the abundance, distribution, and biological characteristics of groundfish resources. Slope species of the deep-water complex (DWC) are of particular importance and include Dover sole, Microstomus pacificus; sablefish, Anoplopoma fimbria; shortspine thornyhead, Sebastolobus alascanus; and longspine thornyhead, S. altivelis. In the fall of 1994, we conducted an experimental gear research cruise in lieu of our normal survey because of concerns about the performance of the survey trawl. The experiment was conducted on a soft mud bottom at depths of 460–490 m off the central Oregon coast. Treatments included different combinations of door-bridle rigging, groundgear weight, and scope length. The experimental design was a 2 ´ 2 ´ 2 factorial within a randomized complete-block. Analysis of variance was used to examine the effects of gear modifications on the engineering performance of the trawl (i.e. trawl dimensions, variation in trawl dimensions, and door attitude) and to determine if catch rates in terms of weight and number of DWC species and invertebrates were affected by the gear modifications. Trawl performance was highly variable for the historically used standard trawl configuration. Improvements were observed with the addition of either a 2-bridle door or lighter ground gear. Changes in scope length had relatively little effect on trawl performance. The interaction of door bridle and ground gear weight had the most effect on trawl performance. In spite of the standard trawl’s erratic performance, catch rates of all four DWC species and invertebrates were not significantly different than the 2-bridle/heavy combination, which did the best in terms of engineering performance. The most important factor affecting DWC catch rates was ground gear. Scope length and the type of door bridle had little effect on DWC catch rates. Subsequent revisions to survey gear and towing protocol and their impact on the continuity of the slope survey time series are discussed.

Gear modifications for fishing octopus, Octopus vulgaris, on live-bottom and adjacent flat bottom habitats in coastal waters off North Carolina

The Common Octopus, Octopus vulgaris, is an r-selected mollusk found off the coast of North Carolina that interests commercial fishermen because of its market value and the cost-effectiveness of unbaited pots that can catch it. This study sought to: 1) determine those gear and environmental factors that influenced catch rates of octopi, and 2) evaluate the feasibility of small-scale commercial operations for this species. Pots were fished from August 2010 through September 2011 set in strings over hard and sandy bottom in waters from 18 to 30 m deep in Onslow Bay, N.C. Three pot types were fished in each string; octopus pots with- and without lids, and conch pots. Proportional catch was modeled as a function of gear design and environmental factors (location, soak time, bottom type, and sea surface water temperature) using binomially distributed generalized linear models (GLM’s); parsimony of each GLM was assessed with Akaike Information Criteria (AIC). A total of 229 octopi were caught throughout the study. Pots with lids, pots without lids, and conch pots caught an average of 0.15, 0.17, and 0.11 octopi, respectively, with high variability in catch rates for each pot type. The GLM that best fit the data described proportional catch as a function of sea surface temperature, soak time, and station; greatest proportional catches occurred over short soak times, warmest temperatures, and less well known reef areas. Due to operating expenses (fuel, crew time, and maintenance), low catch rates of octopi, and high gear loss, a directed fishery for this species is not economically feasible at the catch rates found in this study. The model fitting to determine factors most influential on catch rates should help fishermen determine seasons and gear soak times that are likely to maximize catch rates. Potting for octopi may be commercially practical as a supplemental activity when targeting demersal fish species that are found in similar habitats and depth ranges in coastal waters off North Carolina.

A comparison between the catching efficiency of two milkfish fry collecting gears and their respective modifications

An experiment was conducted along the shore of Culasi, Antique in Panay to compare the milkfry (Chanos chanos) catching efficiency of the ordinary fry seine against its innovation and the ordinary sweeper against its 2 modifications. Results show that it is possible to replace the wings of the presently used sweeper and the ends of the fry seine with a coarse-meshed netting. This improvement decreases the water resistance of the gears and thus enables fry gatherers to use larger ones thereby giving more catch.

Modifications in bangus culture

The article presents the milkfish aquaculture techniques in the Philippines modified from the traditional method. These are modular method and silo method, methods on eliminating snails, fertilizer-water replenishment scheme, supplemental feeding and the stunting of fingerlings are also presented.