Residue specific contributions to stability and activity inferred from saturation mutagenesis and deep sequencing

Multiple methods currently exist for rapid construction and screening of single-site saturation mutagenesis (SSM) libraries in which every codon or nucleotide in a DNA fragment is individually randomized. Nucleotide sequences of each library member before and after screening or selection can be obtained through deep sequencing. The relative enrichment of each mutant at each position provides information on its contribution to protein activity or ligand-binding under the conditions of the screen. Such saturation scans have been applied to diverse proteins to delineate hot-spot residues, stability determinants, and for comprehensive fitness estimates. The data have been used to design proteins with enhanced stability, activity and altered specificity relative to wild-type, to test computational predictions of binding affinity, and for protein model discrimination. Future improvements in deep sequencing read lengths and accuracy should allow comprehensive studies of epistatic effects, of combinational variation at multiple sites, and identification of spatially proximate residues.

Iodide Retention by Modified Kaolinite in the Context of Safe Disposal of High Level Nuclear Waste

Bentonite clay is identified as potential buffer in deep geological repositories (DGR) that store high level radioactive wastes (HLW) as the expansive clay satisfies the expected mechanical and physicochemical functions of the buffer material. In the deep geological disposal of HLW, iodine-129 is one of the significant nuclides, attributable to its long half-life (half life 1⁄4 1:7 × 107 years). However, the negative charge on the basal surface of bentonite particles precludes retention of iodide anions. To render the bentonite effective in retaining hazardous iodide species in DGR, improvement of the anion retention capacity of bentonite becomes imperative. The iodide retention capac-ity of bentonite is improved by admixing 10 and 20% Ag-kaolinite (Ag-K) with bentonite (B) on a dry mass basis. The present study produced Ag-kaolinite by heating silver nitrate-kaolinite mixes at 400°C. Marginal release of iodide retained by Ag-kaolinite occurred under extreme acidic (pH 1⁄4 2:5) and alkaline (pH 1⁄4 12:5) conditions. The swell pressure and iodide etention results of the B-Ag-K specimens bring out that mixing Ag-K with bentonite does not chemically modify the expansive clay; the mixing is physical in nature and Ag-K presence only contributes to iodide retention of the admixture. DOI: 10.1061/(ASCE)HZ.2153-5515.0000121. © 2012 American Society of Civil Engineers.

Characterizing the pocketome of Mycobacterium tuberculosis and application in rationalizing polypharmacological target selection

Polypharmacology is beginning to emerge as an important concept in the field of drug discovery. However, there are no established approaches to either select appropriate target sets or design polypharmacological drugs. Here, we propose a structural-proteomics approach that utilizes the structural information of the binding sites at a genome-scale obtained through in-house algorithms to characterize the pocketome, yielding a list of ligands that can participate in various biochemical events in the mycobacterial cell. The pocket-type space is seen to be much larger than the sequence or fold-space, suggesting that variations at the site-level contribute significantly to functional repertoire of the organism. All-pair comparisons of binding sites within Mycobacterium tuberculosis (Mtb), pocket-similarity network construction and clustering result in identification of binding-site sets, each containing a group of similar binding sites, theoretically having a potential to interact with a common set of compounds. A polypharmacology index is formulated to rank targets by incorporating a measure of druggability and similarity to other pockets within the proteome. This study presents a rational approach to identify targets with polypharmacological potential along with possible drugs for repurposing, while simultaneously, obtaining clues on lead compounds for use in new drug-discovery pipelines.

Structure, stability and elasticity of DNA nanotubes

DNA nanotubes are tubular structures composed of DNA crossover molecules. We present a bottom up approach for the construction and characterization of these structures. Various possible topologies of nanotubes are constructed such as 6-helix, 8-helix and tri-tubes with different sequences and lengths. We have used fully atomistic molecular dynamics simulations to study the structure, stability and elasticity of these structures. Several nanosecond long MD simulations give the microscopic details about DNA nanotubes. Based on the structural analysis of simulation data, we show that 6-helix nanotubes are stable and maintain their tubular structure; while 8-helix nanotubes are flattened to stabilize themselves. We also comment on the sequence dependence and the effect of overhangs. These structures are approximately four times more rigid having a stretch modulus of similar to 4000 pN compared to the stretch modulus of 1000 pN of a DNA double helix molecule of the same length and sequence. The stretch moduli of these nanotubes are also three times larger than those of PX/JX crossover DNA molecules which have stretch moduli in the range of 1500-2000 pN. The calculated persistence length is in the range of a few microns which is close to the reported experimental results on certain classes of DNA nanotubes.

GHG footprint of major cities in India

Concentration of greenhouse gases (GHG) in the atmosphere has been increasing rapidly during the last century due to ever increasing anthropogenic activities resulting in significant increases in the temperature of the Earth causing global warming. Major sources of GHG are forests (due to human induced land cover changes leading to deforestation), power generation (burning of fossil fuels), transportation (burning fossil fuel), agriculture (livestock, farming, rice cultivation and burning of crop residues), water bodies (wetlands), industry and urban activities (building, construction, transport, solid and liquid waste). Aggregation of GHG (CO2 and non-CO2 gases), in terms of Carbon dioxide equivalent (CO(2)e), indicate the GHG footprint. GHG footprint is thus a measure of the impact of human activities on the environment in terms of the amount of greenhouse gases produced. This study focuses on accounting of the amount of three important greenhouses gases namely carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) and thereby developing GHG footprint of the major cities in India. National GHG inventories have been used for quantification of sector-wise greenhouse gas emissions. Country specific emission factors are used where all the emission factors are available. Default emission factors from IPCC guidelines are used when there are no country specific emission factors. Emission of each greenhouse gas is estimated by multiplying fuel consumption by the corresponding emission factor. The current study estimates GHG footprint or GHG emissions (in terms of CO2 equivalent) for Indian major cities and explores the linkages with the population and GDP. GHG footprint (Aggregation of Carbon dioxide equivalent emissions of GHG's) of Delhi, Greater Mumbai, Kolkata, Chennai, Greater Bangalore, Hyderabad and Ahmedabad are found to be 38,633.2 Gg, 22,783.08 Gg, 14,812.10 Gg, 22,090.55 Gg, 19,796.5 Gg, 13,734.59 Gg and 91,24.45 Gg CO2 eq., respectively. The major contributors sectors are transportation sector (contributing 32%, 17.4%, 13.3%, 19.5%, 43.5%, 56.86% and 25%), domestic sector (contributing 30.26%, 37.2%, 42.78%, 39%, 21.6%, 17.05% and 27.9%) and industrial sector (contributing 7.9%, 7.9%, 17.66%, 20.25%, 1231%, 11.38% and 22.41%) of the total emissions in Delhi, Greater Mumbai, Kolkata, Chennai, Greater Bangalore, Hyderabad and Ahmedabad, respectively. Chennai emits 4.79 t of CO2 equivalent emissions per capita, the highest among all the cities followed by Kolkata which emits 3.29 t of CO2 equivalent emissions per capita. Also Chennai emits the highest CO2 equivalent emissions per GDP (2.55 t CO2 eq./Lakh Rs.) followed by Greater Bangalore which emits 2.18 t CO2 eq./Lakh Rs. (C) 2015 Elsevier Ltd. All rights reserved.

Embodied and operational energy of urban residential buildings in India

Globally, buildings consume nearly half of the total energy produced, and consequently responsible for a large share of CO2 emissions. A building's life cycle energy (LCE) comprises its embodied energy (EE) and operational energy (OE). The building design, prevalent climatic conditions and occupant behaviour primarily determines its LCE. Thus, for the identification of appropriate emission-reduction strategies, studies into building LCE are crucial. While OE reflects the energy utilized in operating a, EE comprises the initial capital energy involved in its construction (material and burden associated with material consumption in buildings. Assessment of EE and OE in buildings is crucial towards identifying appropriate design and operational strategies for reduction of the building's life cycle energy. This paper discusses EE and OE assessment of a few residential buildings in different climatic locations in India. The study shows that share of OE and EE in LCE greatly depends upon the types of materials used in construction and extent of space conditioning adopted. In some cases EE can exceed life cycle OE. Buildings with reinforced concrete frame and monolithic reinforced concrete walls have very high EE. (C) 2015 Elsevier B.V. All rights reserved.

Summary report on water quantity and quality studies of Vancouver Lake, Washington

Vancouver Lake, located adjacent to the Columbia River and just north of the Vancouver-Portland metropolitan area, is a "dying" lake. Although all lakes die naturally in geologic time through the process of eutrophication,* Vancouver Lake is dying more rapidly due to man's activities and due to the resultant increased accumulation of sediment, chemicals, and wastes. Natural eutrophication takes thousands of years, whereas man-made modifications can cause the death of a lake in decades. Vancouver Lake does, however, have the potential of becoming a valuable water resource asset for the area, due particularly to its location near the Columbia River which can be used as a source of "flushing" water to improve the quality of Vancouver Lake. (Document pdf contains 59 pages) Community interest in Vancouver Lake has waxed and waned. Prior to World War II, there were relatively few plans for discussions about the Lake and its surrounding land area. A plan to drain the Lake for farming was prohibited by the city council and county commissioners. Interest increased in 1945 when the federal government considered developing the Lake as a berthing harbor for deactivated ships at which time a preliminary proposal was prepared by the City. The only surface water connection between Vancouver Lake and the Columbia River, except during floods, is Lake River. The Lake now serves as a receiving body of water for Lake River tidal flow and surface flow from creeks and nearby land areas. Seasonally, these flows are heavily laden with sediment, septic tank drainage, fertilizers and drainage from cattle yards. Construction and gravel pit operations increase the sediment loads entering the Lake from Burnt Bridge Creek and Salmon Creek (via Lake River by tidal action). The tidal flats at the north end of Vancouver Lake are evidence of this accumulation. Since 1945, the buildup of sediment and nutrients created by man's activities has accelerated the growth of the large water plants and algae which contribute to the degeneration of the Lake. Flooding from the Columbia River, as in 1968, has added to the deposition in Vancouver Lake. The combined effect of these human and natural activities has changed Vancouver Lake into a relatively useless body of shallow water supporting some wildlife, rough fish, and shallow draft boats. It is still pleasant to view from the hills to the east. Because precipitation and streamflow are the lowest during the summer and early fall, water quantity and quality conditions are at their worst when the potential of the Lake for water-based recreation is the highest. Increased pollution of the Lake has caused a larger segment of the community to become concerned. Land use and planning studies were undertaken on the Columbia River lowlands and a wide variety of ideas were proposed for improving the quality of the water-land environment in order to enhance the usefulness of the area. In 1966, the College of Engineering Research Division at Washington State University (WSU0 in Pullman, Washington, was contacted by the Port of Vancouver to determine possible alternatives for restoring Vancouver Lake. Various proposals were prepared between 1966 and 1969. During the summer and fall of 1967, a study was made by WSU on the existing water quality in the Lake. In 1969, the current studies were funded to establish a data base for considering a broad range of alternative solutions for improving the quantity and quality of Vancouver Lake. Until these studies were undertaken, practically no data on a continuous nature were available on Vancouver Lake, Lake River, or their tributaries. (Document pdf contains 59 pages)

Coral Remote Sensing Workshop: Proceedings and Recommendations, 17-18 September, Sheraton, Brickell Ave, Miami FL

Coral reefs exist in warm, clear, and relatively shallow marine waters worldwide. These complex assemblages of marine organisms are unique, in that they support highly diverse, luxuriant, and essentially self-sustaining ecosystems in otherwise nutrient-poor and unproductive waters. Coral reefs are highly valued for their great beauty and for their contribution to marine productivity. Coral reefs are favorite destinations for recreational diving and snorkeling, as well as commercial and recreational fishing activities. The Florida Keys reef tract draws an estimated 2 million tourists each year, contributing nearly $800 million to the economy. However, these reef systems represent a very delicate ecological balance, and can be easily damaged and degraded by direct or indirect human contact. Indirect impacts from human activity occurs in a number of different forms, including runoff of sediments, nutrients, and other pollutants associated with forest harvesting, agricultural practices, urbanization, coastal construction, and industrial activities. Direct impacts occur through overfishing and other destructive fishing practices, mining of corals, and overuse of many reef areas, including damage from souvenir collection, boat anchoring, and diver contact. In order to protect and manage coral reefs within U.S. territorial waters, the National Oceanic and Atmospheric Administration (NOAA) of the U.S. Department of Commerce has been directed to establish and maintain a system of national marine sanctuaries and reserves, and to monitor the condition of corals and other marine organisms within these areas. To help carry out this mandate the NOAA Coastal Services Center convened a workshop in September, 1996, to identify current and emerging sensor technologies, including satellite, airborne, and underwater systems with potential application for detecting and monitoring corals. For reef systems occurring within depths of 10 meters or less (Figure 1), mapping location and monitoring the condition of corals can be accomplished through use of aerial photography combined with diver surveys. However, corals can exist in depths greater than 90 meters (Figure 2), well below the limits of traditional optical imaging systems such as aerial or surface photography or videography. Although specialized scuba systems can allow diving to these depths, the thousands of square kilometers included within these management areas make diver surveys for deeper coral monitoring impractical. For these reasons, NOAA is investigating satellite and airborne sensor systems, as well as technologies which can facilitate the location, mapping, and monitoring of corals in deeper waters. The following systems were discussed as having potential application for detecting, mapping, and assessing the condition of corals. However, no single system is capable of accomplishing all three of these objectives under all depths and conditions within which corals exist. Systems were evaluated for their capabilities, including advantages and disadvantages, relative to their ability to detect and discriminate corals under a variety of conditions. (PDF contains 55 pages)

Sexual Reproduction and Early Development in the Estuarine Sea Anemone, Nematostella vectensis Stephenson, 1935

This dissertation: 1) determines the factor(s) responsible for spawning induction in NematosteJla vectensis; 2) isolates, describes, and documents the source of jelly from egg masses of N. vectensis; and 3) describes N. vectensis' early development. Namatostella vectensis were maintained on a 7-day mussel feeding/water change regime over 159 days. Within 36 hours of mussel feeding/water change. 69.1% of females and 78.5% of males spawned reliably. Through manipulation of feeding, water change, oxygen and nitrogenous waste concentrations, spawning induction was found to be triggered by the oxygen concentration associated with water change, and not by feeding. Ammonia, anemones' major waste product, inhibited this induction in a concentration-dependent manner. Female N. vectensis release eggs in a persistent jellied egg mass which is unique among the Actiniaria. The major component of this egg mass jelly was a positive periodic acid-Schiffs staining, 39.5-40.5 kD glycoprotein. Antibodies developed in rabbits against this glycoprotein bound to jelly of intact egg masses and to granules (~ 2.8 IJm in diameter) present in female anemone mesenteries and their associated filaments. Antibodies did not label male tissues. Nematostella vecfensis embryos underwent first karyokinesis -60 minutes following the addition of sperm to eggs. Second nuclear division took place, followed by first cleavage, 90-120 minutes later. Each of the 4 blastomeres that resulted from first cleavage contained a single nucleus. Arrangement of these blastomeres ranged from radial to pseudospiral. Embryonic development was both asynchronous and holoblastic. Following formation of the 4-cell stage, 71% of embryos proceeded to cleave again to form an 8-cell stage. In each of the remaining 29% of embryos, a fusion of from 2-4 blastomeres resulted in 4 possible patterns which had no affect on either cleavage interval timing or subsequent development. The fusion event was not due to ooplasmic segregation. Blastomeres isolated from 4-celled embryos were regulative and developed into normal planula larvae and juvenile anemones that were 1/4 the size of those that developed from intact 4-celled embryos. Embryos exhibiting the fusion phenomenon were examined at the fine structural level. The fusion phenomenon resulted in formation of a secondary syncytium and was not a mere compaction of blastomeres.

Image-based delineation and classification of built heritage masonry

[EN]Fundación Zain is developing new built heritage assessment protocols. The goal is to objectivize and standardize the analysis and decision process that leads to determining the degree of protection of built heritage in the Basque Country. The ultimate step in this objectivization and standardization effort will be the development of an information and communication technology (ICT) tool for the assessment of built heritage. This paper presents the ground work carried out to make this tool possible: the automatic, image-based delineation of stone masonry. This is a necessary first step in the development of the tool, as the built heritage that will be assessed consists of stone masonry construction, and many of the features analyzed can be characterized according to the geometry and arrangement of the stones. Much of the assessment is carried out through visual inspection. Thus, this process will be automated by applying image processing on digital images of the elements under inspection. The principal contribution of this paper is the automatic delineation the framework proposed. The other contribution is the performance evaluation of this delineation as the input to a classifier for a geometrically characterized feature of a built heritage object. The element chosen to perform this evaluation is the stone arrangement of masonry walls. The validity of the proposed framework is assessed on real images of masonry walls.

Long-Term Evolution Of Delayed Ettringite And Gypsum In Portland Cement Mortars Under Sulfate Erosion

The magnitude evolution of ettringite and gypsum in hydrated Portland cement mortars due to sulfate attack was detected by X-ray powder diffraction. The influences of sulfate concentration and water-to-cement ratio on the evolution of ettringite and gypsum were investigated. Experimental results show that the magnitude of ettringite formation in sodium sulfate solution follows a three-stage process, namely, the 'penetration period', 'enhance period of strength', and 'macro-crack period'. The cracking of concrete materials is mainly attributed to the effect of ettringite. The gypsum formations occurred in two stages, the 'latent period' and the 'accelerated period'. The gypsum formation including ettringite formation was relative to the linear expansion of mortars to some extend. Both water-to-cement ratio and sulfate concentration play important roles in the evolution of ettringite and gypsum. (C) 2008 Elsevier Ltd. All rights reserved.

Aspects of the fishing industry and an overview of artificial reefs and fish aggregating devices for increasing fisheries output and viability in Nigeria

Aspects of the Nigerian fishing industry are outlined to explain the concept of fishing systems viability which is often influenced by a combination of factors including biological productivity, as well as technical, economic and social factors. The productivity of the aquatic environments can be increased by the construction and installation of artificial reefs and fish aggregating devices. These man-made structures provide shelters, food and breeding grounds for fin fish and shell fish. The habitat enhancement techniques are appropriate, efficient, cheap and simple strategic options for increase in fish production. Recommendations for effective utilization and long term management are outlined.

Lake Kainji water hyacinth infestation and the control strategies

The paper documents the aquatic vascular plants situation of Lake Kainji, particularly with the invasion of water hyacinth (Eichhormia crassipes) hitherto alien to the lake system. The frequency of occurrence of some aquatic plants on Lake Kainji have increased whilst a few decreased over the period between 1984 and 1995. More than 53.6% of the lake surface has now been covered by aquatic vascular plants of which water hyacinth alone covered 30.6% (16.4% of the system). Apart from the characteristic rapid multiplication and biomass build-up of the plant, more than 9.1 clumps with a diameter range between 0.8-4cm enter the lake per minute. Thus more than 16.4ha per day or annually 6000ha of water hyacinth are added as drifts. Because of the peculiar hydrological cycle of the lake and the plant aggressiveness and invasiveness, NIFFR (with the support of the GTZ) had developed an approach of control which combines manual, biological, natural, boom construction and chemical measures. These are discussed

The optoelectronic swept-frequency laser and its applications in ranging, three-dimensional imaging, and coherent beam combining of chirped-seed amplifiers

This thesis explores the design, construction, and applications of the optoelectronic swept-frequency laser (SFL). The optoelectronic SFL is a feedback loop designed around a swept-frequency (chirped) semiconductor laser (SCL) to control its instantaneous optical frequency, such that the chirp characteristics are determined solely by a reference electronic oscillator. The resultant system generates precisely controlled optical frequency sweeps. In particular, we focus on linear chirps because of their numerous applications. We demonstrate optoelectronic SFLs based on vertical-cavity surface-emitting lasers (VCSELs) and distributed-feedback lasers (DFBs) at wavelengths of 1550 nm and 1060 nm. We develop an iterative bias current predistortion procedure that enables SFL operation at very high chirp rates, up to 10^16 Hz/sec. We describe commercialization efforts and implementation of the predistortion algorithm in a stand-alone embedded environment, undertaken as part of our collaboration with Telaris, Inc. We demonstrate frequency-modulated continuous-wave (FMCW) ranging and three-dimensional (3-D) imaging using a 1550 nm optoelectronic SFL.

We develop the technique of multiple source FMCW (MS-FMCW) reflectometry, in which the frequency sweeps of multiple SFLs are "stitched" together in order to increase the optical bandwidth, and hence improve the axial resolution, of an FMCW ranging measurement. We demonstrate computer-aided stitching of DFB and VCSEL sweeps at 1550 nm. We also develop and demonstrate hardware stitching, which enables MS-FMCW ranging without additional signal processing. The culmination of this work is the hardware stitching of four VCSELs at 1550 nm for a total optical bandwidth of 2 THz, and a free-space axial resolution of 75 microns.

We describe our work on the tomographic imaging camera (TomICam), a 3-D imaging system based on FMCW ranging that features non-mechanical acquisition of transverse pixels. Our approach uses a combination of electronically tuned optical sources and low-cost full-field detector arrays, completely eliminating the need for moving parts traditionally employed in 3-D imaging. We describe the basic TomICam principle, and demonstrate single-pixel TomICam ranging in a proof-of-concept experiment. We also discuss the application of compressive sensing (CS) to the TomICam platform, and perform a series of numerical simulations. These simulations show that tenfold compression is feasible in CS TomICam, which effectively improves the volume acquisition speed by a factor ten.

We develop chirped-wave phase-locking techniques, and apply them to coherent beam combining (CBC) of chirped-seed amplifiers (CSAs) in a master oscillator power amplifier configuration. The precise chirp linearity of the optoelectronic SFL enables non-mechanical compensation of optical delays using acousto-optic frequency shifters, and its high chirp rate simultaneously increases the stimulated Brillouin scattering (SBS) threshold of the active fiber. We characterize a 1550 nm chirped-seed amplifier coherent-combining system. We use a chirp rate of 5*10^14 Hz/sec to increase the amplifier SBS threshold threefold, when compared to a single-frequency seed. We demonstrate efficient phase-locking and electronic beam steering of two 3 W erbium-doped fiber amplifier channels, achieving temporal phase noise levels corresponding to interferometric fringe visibilities exceeding 98%.

Observational and theoretical advances in cosmological foreground emission

Observational and theoretical work towards the separation of foreground emission from the cosmic microwave background is described. The bulk of this work is in the design, construction, and commissioning of the C-Band All-Sky Survey (C-BASS), an experiment to produce a template of the Milky Way Galaxy's polarized synchrotron emission. Theoretical work is the derivation of an analytical approximation to the emission spectrum of spinning dust grains.

The performance of the C-BASS experiment is demonstrated through a preliminary, deep survey of the North Celestial Pole region. A comparison to multiwavelength data is performed, and the thermal and systematic noise properties of the experiment are explored. The systematic noise has been minimized through careful data processing algorithms, implemented both in the experiment's Field Programmable Gate Array (FPGA) based digital backend and in the data analysis pipeline. Detailed descriptions of these algorithms are presented.

The analytical function of spinning dust emission is derived through the application of careful approximations, with each step tested against numerical calculations. This work is intended for use in the parameterized separation of cosmological foreground components and as a framework for interpreting and comparing the variety of anomalous microwave emission observations.