ALBERT

Description: Diffuse nutrient pollution from agriculture has been the concern of policymakers for several decades, and yet it remains a persistent environmental issue. The current approach to mitigating the problem is predominantly via command and control regulation within the Nitrates Directive and the Water Framework Directive. This article will set out how diffuse pollution can be considered a wicked policy problem which acts as an explanation of how it has eluded the current regulatory regime. It will further establish that the traditional planning process overlooked the complexity of the problem. Finally, it will illustrate the ineffectiveness of the current regulatory framework to mitigate the problem. This will be exemplified through the legal framework of Northern Ireland.

Description: This analysis explores new developments in judicial review of planning policy interpretation. It shows how the nature of policy, often contextual and judgment-dependent, has led the UK Supreme Court to rethink the standard of review applicable to this issue. By considering the recent decision in Samuel Smith as part of a trilogy of cases—including Tesco Stores and Hopkins Homes—this analysis reveals a change in judicial attitudes, away from the expansive judicial supervision upheld in Tesco Stores. Furthermore, this study reflects on how this change is related to two wider ideas. The first is the Court’s understanding of the law and policy divide in the planning field, whilst the second is to do with a pragmatic stance regarding the purpose of the planning system and the institutional role of the courts in it. Finally, this analysis shows how the new approach emphasises the distinctive character of policy in the planning context.

Description: In the present work, the performance of carbon nanotubes (c-CNTs) functionalized polydimethylsiloxane (PDMS) based coatings as extractive phases for in-tube solid phase microextraction (IT-SPME) coupled to Capillary LC (CapLC) has been evaluated. Carboxylic-single walled carbon nanotubes (c-SWNTs) and carboxylic-multi walled carbon nanotubes (c-MWNTs) have been immobilized on the activated surface of PDMS capillary columns. The effect of different percentages of diphenyl groups in the PDMS extractive phase has also been evaluated. The extraction capability of the capillary columns has been tested for different organic pollutants, nitrogen heterocyclic compounds and polycyclic aromatic compounds (PAHs). The results indicated that the use of the c-CNTs-PDMS capillary columns improve pyriproxyfen and mainly PAH extraction. Triazines were better extracted by unmodified TRB-35 and modified c-CNTs-PDMSTRB-5. The results showed that the extraction capability of the c-CNT capillary columns depends not only on the polarity of the analytes (as it occurs with PDMS columns) but also on the interactions that the analytes can establish with the immobilized c-CNTs on the PDMS columns. The extraction efficiency has been evaluated on the basis of the preconcentration rate that can be achieved, and, in this sense, the best c-CNTs-PDMS capillary column for each group of compounds can be proposed.

Description: The Saudi Arabia (SA) climate varies greatly, depending on the geography and the season. According to K ppen and Geiger, the climates of SA is “desert climate”. The analysis of the seasonal rainfall detects that spring and winter seasons have the highestrainfall incidence, respectively. Through the summer,small quantities of precipitation are observed, while autumn received more precipitation more than summer season considering the total annual rainfall. In all seasons, the SW area receives rainfall, with a maximum in spring, whereas in the summer season, the NE and NW areas receive very little quantities of precipitation. The Rub Al-Khali (the SE region) is almost totally dry. The maximum amount of annual rainfall does not always happen at the highest elevation. Therefore, the elevation is not the only factor in rainfall distribution.A great inter-annual change in the rainfall over the SA for the period (1978–2009) is observed. In addition, in the same period, a linear decreasing trend is found in the observed rainfall, whilst in the recent past (1994–2009) a statistically significant negative trend is observed. In the Southern part of the Arabian Peninsula (AP) and along the coast of the Red Sea, it is interesting to note that rainfall increased, whilst it decreased over most areas of SA during the 2000–2009 decade, compared to 1980–1989.Statistical and numerical models are used to predict rainfall over Saudi Arabia (SA). The statistical models based on stochastic models of ARIMA and numerical models based on Providing Regional Climates for Impact Studies of Hadley Centre (PRECIS). Climate and its qualitative character and quantified range of possible future changes are investigated. The annual total rainfall decreases in most regions of the SA and only increases in the south. The summertime precipitation will be the highest between other seasons over the southern, the southwestern provinces and Asir mountains, while the wintertime rainfall will remain the lowest.The climate in the SA is instructed by the El Niño Southern Oscillation (ENSO) and other circulations such as centers of high and low pressure, the North Atlantic Oscillation (NAO) and SOI. Strength and oscillation of subtropical jet stream play a big role in pulling hot, dry air masses of SA.

Description: Natural gas is an extremely important bridge fuel to a low-carbon energy economy for improving local air quality. Coal to synthetic natural gas (SNG) is an effective way to convert the high-carbon energy (coal) into the low-carbon energy with rich hydrogen (natural gas). For the modern coal to SNG industry, the high-temperature methanation (HTM) catalyst plays an important role, and the advanced evaluation process should necessitate the elimination of mass transfer effect. Some simple but effective model catalysts, such as slab and sphere, can be very helpful in defining the reaction conditions, and thus facilitating the evaluation process for real HTM catalysts. In this work, slab and sphere model catalysts were adopted to derive mass transfer and reaction-coupled equations, the numerical methods were used to solve the coupled equations for the concentration profiles in catalysts, and the effectiveness factors were accordingly calculated. By taking advantage of the Thiele module and the Weisz–Prater module , the criteria for the elimination of mass transfer effect in the HTM catalyst evaluation process were successfully defined. This work also complements the Weisz–Prater criterion by incorporating ‘negative reaction orders’.

Description: The energy expended for conditioning buildings around the world is worryingly large and increasing every year. Currently, almost half of houses around the world use some type of energy-expensive conventional air-conditioning system. These systems are high emitters of gases such as carbon dioxide and so high contributors to climate change. Consequently, alternatives must be considered. Earth–air heat exchangers (EAHEs) and phase-change materials (PCMs) may be options; they have, however, limitations. This paper proposes a novel hybrid space-conditioning system combining EAHEs with PCMs, which uses surfaces as sources of heating or cooling to provide better temperature distribution across a space and comfort enhancement with low energy use. The idea is to use an EAHE to provide cool air to discharge the PCM, consequently helping to overcome most of the limitations of both strategies. Two of a series of experiments undertaken to test the proposed system are reported in this article. The EAHE + PCM system compared with a reference room could decrease temperature swings by up to 47%. The system was proved to work although further work is required to make it commercially viable.

Description: The design of a future building with very high-energy efficiency demands from the architect to study the available solar resources in this urban environment. The purpose of the presented methodology is to study the variations in all components of the incident solar radiation daily, monthly and seasonally for all building facades. This is realized in the computer program 3D–SOLARIA. In the focus of the paper is the estimation of the background component of the incident diffuse solar irradiation on a building facade under orthogonally obstructed sky, using anisotropic sky view factors.

Description: The assessment of building envelope sustainability using existing building performance assessment methods is still a challenge and yet to be fully addressed. This is due to the single-dimensional nature and lack of integration of sustainable performance values. Besides, the issue of sustainability assessment in the building envelope requires considering many factors including life cycle considerations. As such, in an effort to develop an integrated approach that combines relevant sustainable development factors and life cycles, an integrated performance model (IPM) was developed. The IPM is an essential tool developed to aid the sustainable design of the residential building envelope that can reduce carbon emission and whole residential building energy consumption and ensure sustainable performance of the building envelope.

Description: The results and considerations on one self-consumption photovoltaic installation with net balance in Granada (South of Spain) are presented and discussed in this paper. The use of the building (one faculty) makes it optimal for this kind of consumption. Finally, the potential benefits and problems of self-consumption with net balance are presented.

Description: Passive convective heat transfer enhancement can be achieved by improving the thermo-physical properties of the working fluid, changing flow geometry or both. This work presents a numerical study to investigate the combined effect of using helical coils and nanofluids on the heat transfer characteristics and pressure losses in turbulent flow regime. The developed computational fluid dynamics models were validated against published experimental data and empirical correlations. Results have shown that combining the effects of alumina (Al 2 O 3 ) nanoparticles and tube coiling could enhance the heat transfer coefficient by up to 60% compared with that of pure water in straight tube at the same Reynolds number. Also, results showed that the pressure drop in helical coils using Al 2 O 3 nanofluid for volume fraction of 3% was six times that of water in straight tubes (80% of the pressure drop increase is due to nanoparticles addition), while the effect of Reynolds number on the pressure drop penalty factor was found to be insignificant.

Description: The demand for better indoor environment has led to a wide use of heating, ventilating and air conditioning (HVAC) systems. However, this demanding cannot be sustained without a strategy to reduce the energy consumption of HVAC systems. Employing advanced HVAC control is one of the strategies to maintain high-quality indoor thermal comfort with low energy consumption. This paper reviews the development of control technologies for HVAC systems. It summarizes the problems in existing reviews and gives a future perspective on HVAC control systems for occupant comfort and energy efficiency.

Description: The use of a novel micro pressurized liquid extraction (µPLE) method for the isolation of 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) from various solid samples is explored. The technique employs rapid heating in a single static extraction mode to remove analytes in a matter of seconds from 5–10 mg samples using only 125 µL of solvent. For example, results show that 30 s extractions with toluene at 200 °C produce respective PAH recovery ranges of 90%–130% and 88%–114% from samples of soil and smoked chicken. Comparatively, solids containing significant amounts of biochar were more challenging to extract from. For instance, when using a pure biochar sample matrix, recoveries for the 16 PAHs range from only 33%–66% after 60 s of extraction with toluene at 200 °C. Overall, these extraction results agree very well with those reported when using conventional methods on similar samples. Therefore, the findings indicate that µPLE can potentially provide an alternative sample preparation method for PAHs that is both very rapid and requires little solvent.

Description: Aquifers within the Columbia River Basalt Group (CRBG) provide a critical water supply throughout much of the Pacific Northwest of the United States. Increased pumping has resulted in water level declines in this region. Recharge into this aquifer system is generally not well understood. Recent suggestions of probable decades-long droughts in the 21st century add to this problem. We show that geophysical methods can provide useful parameters for improved modeling of aquifers in a primary CRBG aquifer located on the eastern edge of the Columbia Plateau. Groundwater models depend in part on the area, thickness, porosity, storativity, and nature of confinement of this aquifer, most of which are poorly constrained by existing well information and previous stress tests. We have made use of surface gravity measurements, borehole gravity measurements, barometric efficiency estimates, earth tidal response, and earthquake seismology observations to constrain these parameters. We show that the aquifer, despite its persistent drawdown, receives a great deal of recharge annually. Much of the recharge to the aquifer is due to leakage from overlying flows, ultimately tied to precipitation, an important result for future aquifer management in times of sustained drought.

Description: Low flows of the Ilmenau River (1434 km2) in northwest Germany have decreased by about 25% over the last 50 years. In the same period, moderate climate changes have taken place and annual groundwater abstractions for sprinkler irrigation have increased by up to 50 hm3 (million m3), with a strong variation due to the respective prevailing weather conditions. Time-series analyses with multiple regression analysis allow detecting and quantifying different influences on low flows. It is also shown that farmers allocate irrigation water volumes carefully according to seasonal precipitation and temperatures. Decline of groundwater levels in summer and the low flow situation are aggravated by the cumulative effect of higher irrigation in drier years. Groundwater recharge and recovery of the water table have been observed subsequently during the winter season.

Description: Cities’ energy usage accounts for two thirds of global primary energy consumption. Energy efficiency in urban areas is, therefore, one of the most important topics to consider when dealing with urban sustainability. This paper evaluates the goals for increasing energy efficiency and use of renewable energy sources in the areas of transportation, buildings and consumers’ awareness, as stated in the Climate action plan, for the municipality of Eskilstuna, Sweden. The efforts of the municipality to successfully reach their energy efficiency goals, are described in this paper including future perspectives. The results show that although the municipality counts with the advantage of owning and working together with the local housing company and energy provider, in order to reach the established goals, additional strategies need to be considered. For an increased use of renewable energy sources, analysis of rooftops suitable for photovoltaic (PV) installation should be carried out as well as the integration of goals for self-consumption. In the transport field, the city needs to prepare for large-scale electric vehicle (EV) market penetration and to consider different bike or car sharing options. Finally, more specific awareness campaigns are needed to engage the citizens in reducing their energy consumption and living a more sustainable life.

Description: Drought is a serious natural hazard with far-reaching impacts including soil damages, economic losses, and threatening the livelihood and health of local residents. The goal of the present work was to monitor the vegetation health across Lebanon in 2014 using remote sensing techniques. Landsat images datasets, with a spatial resolution of 30 m and from different platforms, were used to identify the VCI (Vegetation Condition Index) and TCI (Temperature Condition Index). The VCI was based on the Normalized Difference Vegetation Index (NDVI) datasets. The TCI used land surface temperature (LST) datasets. As a result, the VHI (Vegetation Health Index) was produced and classified into five categories: extreme, severe, moderate, mild, and no drought. The results show practically no extreme drought (~0.27 km2) in the vegetated area in Lebanon during 2014. Moderate to severe drought mainly occurred in the north of Lebanon (i.e., the Amioun region and the plain of Akkar). The Tyr region and the Bekaa valley experienced a low level of drought (mild drought). This approach allows decision makers to monitor, investigate and resolve drought conditions more effectively.

Description: Soil water potential (Ψ) controls the dynamics of water in soils and can therefore affect greenhouse gas fluxes. We examined the relationship between soil moisture content (θ) at five different levels of water potential (Ψ = 0, −0.05, −0.1, −0.33 and −15 bar) and greenhouse gas (carbon dioxide, CO2; nitrous oxide, N2O and methane, CH4) fluxes. The study was conducted in 2011 in a silt loam soil at Freeman farm of Lincoln University. Soil samples were collected at two depths: 0–10 and 10–20 cm and their bulk densities were measured. Samples were later saturated then brought into a pressure plate for measurements of Ψ and θ. Soil air samples for greenhouse gas flux analyses were collected using static and vented chambers, 30 cm in height and 20 cm in diameter. Determination of CO2, CH4 and N2O concentrations from soil air samples were done using a Shimadzu Gas Chromatograph (GC-14). Results showed that there were significant correlations between greenhouse gas fluxes and θ held at various Ψ in the 0–10 cm depth of soil group. For instance, θ at Ψ = 0 positively correlated with measured CO2 (p = 0.0043, r = 0.49), N2O (p = 0.0020, r = 0.64) and negatively correlated with CH4 (p = 0.0125, r = −0.44) fluxes. Regression analysis showed that 24%, 41% and 19% of changes in CO2, N2O and CH4 fluxes, respectively, were due to θ at Ψ = 0 (p 〈 0.05). This study stresses the need to monitor soil water potential when monitoring greenhouse gas fluxes.

Description: A device (prototype) with a working volume of 200 L was used to deplete olive mill wastewater (OMW) of polyphenols. The OMW transformed into feedstock by means of the device was then used for feeding a lab-scale photobioreactor, just for testing the production of bioH2. The main novelty of this prototype consists in the combination of several adsorbent matrices and the exploitation of their synergic action. In this investigation, three matrices have been used: active carbon, Azolla and zeolite. The device was operated at an olive oil company located in the heart of the Chianti zone (Province of Florence, Italy). The efficiency of polyphenol removal obtained using the device was ≥96%. The multi-matrix effluent (MMeff) generated was then used to obtain three different culture broths containing 25%, 50% and 100% of MMeff, respectively. The diluted (with water) culture broths were suitable for hydrogen generation, with the highest hydrogen production rate (12.7 mL H2/Lculture/h) being obtained using 50% MMeff. The hydrogen yields were: 334 mL H2/L of MMeff, when feeding the photofermenter with pure effluent (100%); 1308 mL H2/L of MMeff, with the half-diluted effluent (50%, v/v); and 432 mL H2/L of MMeff, with the highest-diluted effluent (25%, v/v).

Description: The microbiological and chemical drinking water quality of 20 vulnerable Finnish small groundwater supplies was studied in relation to environmental risk factors associated with potential sources of contamination. The microbiological parameters analyzed included the following enteric pathogens: Giardia and Cryptosporidium, Campylobacter species, noroviruses, as well as indicator microbes (Escherichia coli, intestinal enterococci, coliform bacteria, Clostridium perfringens, Aeromonas spp. and heterotrophic bacteria). Chemical analyses included the determination of pH, conductivity, TOC, color, turbidity, and phosphorus, nitrate and nitrite nitrogen, iron, and manganese concentrations. Giardia intestinalis was detected from four of the water supplies, all of which had wastewater treatment activities in the neighborhood. Mesophilic Aeromonas salmonicida, coliform bacteria and E. coli were also detected. None of the samples were positive for both coliforms and Giardia. Low pH and high iron and manganese concentrations in some samples compromised the water quality. Giardia intestinalis was isolated for the first time in Finland in groundwater wells of public water works. In Europe, small water supplies are of great importance since they serve a significant sector of the population. In our study, the presence of fecal indicator bacteria, Aeromonas and Giardia revealed surface water access to the wells and health risks associated with small water supplies.

Description: For 20 years, the number of resource policy approaches with direct and indirect relations to raw materials, resource and material efficiency has grown enormously at national and international level. This discussion paper makes an inventory of different political and regulatory approaches that contain a direct or indirect reference to resources such as construction materials, industrial minerals, or metals. They are examined and evaluated regarding foci and resource priorities as well as further categories such as target lines, governance levels, indicators used, integration into wider target systems, specification, and implementation. The aim is to provide an overview of the spectrum of resource objectives in international, European, and national strategies, programs, and initiatives. The closer analysis of raw material targets embedded in the policy programs and legal approaches reveals that most goals lack a time frame and a concrete vision, thus remain at a strategic level. To complement the overview, the state of research in the field of modeling and simulation is briefly discussed. Concluding remarks concerning their relation to the objectives identified and the task of target setting complete the discussion.

Description: In buildings without mechanical ventilation, the fresh air needed by the occupants is supplied only by air infiltration or natural ventilation. Its quantification by analytical methods is difficult, and so experimental methods such as the tracer gas technique are very useful. In this work, several case studies are presented, where one may see the potential of this technique to detect small differences in building air tightness, due to different envelope elements, namely the type of window frames and location of blinds' boxes. The applicability of this technique to other spaces rather than buildings can also be analyzed. When dealing with mechanical ventilation, it is difficult to measure the duct air flow rate precisely, because of the minimum duct length necessary to obtain a good mixing of the tracer gas with the air, when conventional probes are used. Research was carried out with the development of new probes to overcome this situation, and results are also shown. With accurate measurements of air flow rates, it is possible to reduce CO 2 emissions due to air heating or cooling in buildings.

Description: A number of factors influence the performance of an ejector, e.g. working fluid, geometry and operating conditions. In the present work, six low-environmental-impact working fluids were evaluated for their use in an ejector cooling system running on low-temperature thermal energy. The numerical analysis was based on a model applying the 1D constant-pressure mixing theory. Ejector performance was assessed for the temperatures of the generator, evaporator and condenser in the range of 80–120°C, 5–15°C and 25–40°C, respectively. The results indicated that owing to its high coefficient of performance and moderate operating pressures throughout the entire ejector cycle, isobutane is a good choice for a refrigerant. The area ratio required for running the ejector in critical mode, under changing operating conditions, varied in a significant range regardless of the selected refrigerant. This clearly indicates the importance of a variable geometry ejector design to strengthen the position of ejector cooling systems among other refrigeration technologies.

Description: Energy security remains a vital issue for the European Union (EU), even more so in the wake of the events that unfolded in early 2014 in Ukraine. The EU’s already fragile position in the international energy arena in terms of security of supply appears to be more uncertain than ever after its umpteenth fallout with its historic energy supplier, Russia. This situation is untenable and calls for swift and decisive action to adequately tackle the issue once and for all. The article looks at the creation of a single EU energy market through integration of energy networks in the EU. It then examines various ways to diversify the EU’s energy supply, whether through increasing the import of liquefied natural gas, through its relations with the Eurasian Union, the promotion of renewable energy or the construction of alternative pipelines and energy routes. The article then offers an analysis of the latest developments of the Energy Charter Conference. The article concludes that from energy transit, to technology transfer, to investment protection, energy and trade present interplays across various fields. Improvements can be made to the EU trading system to ensure greater energy security and more efficient energy markets.

Description: Despite the Egyptian authorities’ great hopes for the fledging shale gas industry in Egypt, it appears that it could be difficult for Egypt to realize these lofty ambitions, at least in the near future. The Egyptian shale gas industry faces technical challenges as well as a critical lack of detailed regulations regarding the exploration for and the production of shale gas. There is also a lack of guidance over the manner in which foreign investments can be made in the industry.

Description: Heat flow of the sedimentary succession of the Eastern Canada Sedimentary Basins varies from 40 mW/m2 close to the exposed shield in the north to high 60–70 mW/m2 in the southwest–northeast St. Lawrence corridor. As high fluid flow rates are required for a successful geothermal application, the most important targets are deep existing permeable aquifers rather than hard rock, which would need to be fracked. Unfortunately, the ten most populated Québec urban centers are in the areas where the Grenville (Canadian Shield) is exposed or at shallow depths with sedimentary cover where temperatures are 30 °C or less. The city of Drummondville will be the exception, as the basement deepens sharply southwest, and higher temperatures reaching >120 °C are expected in the deep Cambrian sedimentary aquifers near a 4–5-km depth. Deep under the area where such sediments could be occurring under Appalachian nappes, temperatures significantly higher than 140 °C are predicted. In parts of the deep basin, temperatures as high as 80 °C–120 °C exist at depths of 3–4 km, mainly southeast of the major geological boundary: the Logan line. There is a large amount of heat resource at such depths to be considered in this area for district heating.

Description: This research looks at coupling desalination with renewable energy sources to create a high-value product (treated water) from two low value resources (brackish groundwater and intermittent solar energy). Desalination of brackish groundwater is already being considered as a potential new water supply in Texas. This research uses Texas as a testbed for spatially-resolved analysis techniques while considering depth to brackish groundwater, water quality, and solar radiation across Texas to determine the locations with the best potential for integrating solar energy with brackish groundwater desalination. The framework presented herein can be useful for policymakers, regional planners, and project developers as they consider where to site desalination facilities coupled with solar photovoltaics. Results suggest that the northwestern region of Texas—with abundant sunshine and groundwater at relatively shallow depths and low salinity in areas with freshwater scarcity—has the highest potential for solar powered desalination. The range in capacity for solar photovoltaic powered reverse osmosis desalination was found to be 1.56 × 10—6 to 2.93 × 10—5 cubic meters of water per second per square meter of solar panel (m3/s/m2).

Description: Ground source heat pump (GSHP) systems have been proven to have higher efficiency compared to conventional air source heat pump systems for space heating and cooling applications. While vertical ground heat exchangers (GHE) are favorable in GSHP installation, this type of configuration requires higher capital costs as opposed to horizontal configuration. Numerical simulation has been used to accurately predict the thermal performance of GHE. In this paper, numerical analysis of thermal performance for slinky horizontal GHE loops in different orientations and operation modes is discussed. It was found that the loop orientation is not so important due to the little effect it has on thermal performance. While the mean heat exchange rate of copper loop increases 48% compared to HDPE loop, the analysis supports the common claim that heat exchange rate is predominantly limited by the thermal conductivity of the ground. With the same amount of circulation work, the mean heat exchange rate increases by 83%–162% when operated in parallel loops operations. The performance in these operations can be further optimized to 10%–14% increase when spacing between adjacent loops was provided. The spacing helps to minimize interference of heat flow that would penalize the overall thermal performance.

Description: Small Island Developing States (SIDS) of the Pacific over the last decade have established some of the most ambitious renewable energy targets in the world. The promotion of renewable energy has been motivated by a desire to lessen dependence on fossil fuels, given the adverse economic impacts of high oil prices on these countries. Efforts to attract development assistance and to strengthen the position of Pacific SIDS in climate change negotiations have likely also played a role. This paper explores the development of renewable energy resources in the Pacific through a public policy lens. The ambitious renewable energy targets established by Pacific SIDS are argued to be appropriate in some cases, but in other cases are criticised on economic grounds. A potential trade-off is identified between the risk mitigation benefits and poverty alleviation benefits of different renewable technology investments, with questions raised about whether support for the former rather than the latter by development partners is appropriate. A number of institutional and financial challenges to the development of renewable energy resources in Pacific SIDS are also discussed.

Description: The State of Wisconsin is located in an unusually water-rich portion of the world in the western part of the Great Lakes region of North America. This article presents an overview of the major groundwater quantity and quality concerns for this region in a geologic context. The water quantity concerns are most prominent in the central sand plain region and portions of a Paleozoic confined sandstone aquifer in eastern Wisconsin. Water quality concerns are more varied, with significant impacts from both naturally occurring inorganic contaminants and anthropogenic sources. Naturally occurring contaminants include radium, arsenic and associated heavy metals, fluoride, strontium, and others. Anthropogenic contaminants include nitrate, bacteria, viruses, as well as endocrine disrupting compounds. Groundwater quality in the region is highly dependent upon local geology and land use, but water bearing geologic units of all ages, Precambrian through Quaternary, are impacted by at least one kind of contaminant.

Description: Urbanized areas of the southwestern/western United States are among the fastest growing in the nation and face multiple water resource challenges. Low impact development (LID)/green infrastructure (GI) practices are increasingly popular technologies for managing stormwater; however, LID is often not as common in the southwest/west due to the lack of regulatory and/or economic drivers. There is also a lack of performance evaluation of these practices, particularly at the field scale. This study focused on investigating the hydrologic and pollutant removal performance of field-scale LID/GI systems in arid/semi-arid climates. Nine typical practices were reviewed: rainwater harvest system, detention pond, retention pond, bioretention, media filter, porous pavement, vegetated swale/buffer/strip, green roof, and infiltration trench, as well as integrated LIDs. We evaluate these practices by a cost-effectiveness analysis and also recommend best practices for the arid/semi-arid area. The analysis provides data support and insights for future implementation of LID/GI in the southwest/west.

Description: The spatial footprint of unconventional (hydraulic fracturing) and conventional oil and gas development in the Marcellus Shale region of the State of Pennsylvania was digitized from high-resolution, ortho-rectified, digital aerial photography, from 2004 to 2010. We used these data to measure the spatial extent of oil and gas development and to assess the exposure of the extant natural resources across the landscape of the watersheds in the study area. We found that either form of development: (1) occurred in ~50% of the 930 watersheds that defined the study area; (2) was closer to streams than the recommended safe distance in ~50% of the watersheds; (3) was in some places closer to impaired streams and state-defined wildland trout streams than the recommended safe distance; (4) was within 10 upstream kilometers of surface drinking water intakes in ~45% of the watersheds that had surface drinking water intakes; (5) occurred in ~10% of state-defined exceptional value watersheds; (6) occurred in ~30% of the watersheds with resident populations defined as disproportionately exposed to pollutants; (7) tended to occur at interior forest locations; and (8) had >100 residents within 3 km for ~30% of the unconventional oil and gas development sites. Further, we found that exposure to the potential effects of landscape disturbance attributable to conventional oil and gas development was more prevalent than its unconventional counterpart.

Description: Large-size (4–5 µm) superficially porous particles yield lower plate heights (e.g., the minimal reduced plate height or hmin ≈ 1.5) than fully porous particles of a similar size when packed into large-bore columns. This property allows for better chromatographic performance without the higher pressures required for smaller particles. This study explores the use of such particles in microfluidic LC columns where materials and fitting pressure limits can constrain the size of particle used. The theoretically predicted performance improvements compared to fully porous particles were not demonstrated in capillary columns (with hmin ≈ 2 for both particle types), in agreement with previous studies that examined smaller superficially porous particles. Microfluidic columns were then compared to capillary columns. Capillary columns significantly outperformed microfluidic columns due to imperfections imposed by microfluidic channel asymmetry and world-to-chip connection at the optimal flow rate; however, superficially porous particles packed in microfluidic LC columns had flatter plate height versus flow rate curves indicating potential for better performance at high reduced velocities.

Description: The thermal performance of a disc-shaped heat generation body with cooling channels is investigated by using constructal theory based on previous model of heat convection on a disc and previous analytical method of heat conduction on a disc. By taking minimum dimensionless maximum thermal resistance as optimization objective, the optimal aspect ratio of the elemental sector in the radial-pattern disc is obtained for the specified power pumping of the elemental sector; the optimal width ratio of the first-order and elemental cooling channels and the optimal dimensionless radius of the elemental sector are obtained for the specified power pumping of the disc. There exists a critical radius of the disc to determine whether the radial-pattern design and branched-pattern design should be adopted. These conclusions are different from those obtained by Wechsatol et al. 's model, and the essential reason for these differences is that the power pumping is specified in this article, but not the specified flow rate number in Wechsatol et al. 's model. Finally, the assumption about the heat capacity of the coolant and the thermal conductivity of the disc is validated. An analytical method is introduced in this article, which can provide another thought for the constructal optimization of disc with heat convection. The optimal constructs of the discs are obtained for the specified power pumping, which provides some different guidelines for the design of disc with heat convection.

Description: In this paper, solar box cookers with ordinary and finned absorber plates are theoretically investigated in terms of thermodynamic performance parameters. For a typical summer and winter day, temperatures of solar cooker components such as glass cover, internal air, absorber plate, cooking vessel and the liquid are determined theoretically versus time. The analysis is carried out for conventional and finned absorber plates, and accuracy of the results is verified by a previously published experimental work. Energy and exergy efficiencies of box-type solar cookers are plotted versus time for various cases. The methodology presented in this paper enables to make a preliminary evaluation of ordinary and modified solar box cookers with respect to changes in main environmental parameters such as temperature and illumination intensity. Some recommendations are also made to enhance the power outputs of the aforementioned solar cookers.

Description: The sun is the more plentiful source of natural power that we have in the Earth. However, the amount of radiation reaching the Earth's surface depends on astronomical and climatic factors. One of the factors exerting a greater influence is cloudiness. For that reason, it is important to quantify its influence on solar radiation. According to that, in this work, a prototype to capture images of the celestial vault is designed and implemented to be able to measure parameters related to the cloud properties and, later, to determine their influence on solar radiation.

Description: Under transient climatic conditions, solar water heaters using heat pipes are more effective at capturing incident solar radiation than other equivalent sized solar water heaters. The cost must be reduced to improve uptake of such systems. To investigate two methods were considered by this study: thermosyphon fluid flow and reflective concentrators. A physical reconfigurable laboratory model of the manifold and associated condensers of a heat-pipe-evacuated tube system were fabricated; fluid circulation was via thermosyphonic action, particle imaging velocimetry derived velocity maps and the use of concentrators was simulated. When condenser spacing was doubled, the Nusselt number increased by 43%, the velocity by 55% but the heat transfer efficiency of the model manifold decreased by 9%. Potential annual energy savings of 10 207 GWh could be realized if such systems could be successfully fabricated.

Description: Although globotetraosylceramide (Gb4) is only recognized by a single member of the verotoxin family namely, the pig edema disease toxin (VT2e), removal of the acetyl group from the terminal N-acetyl hexosamine of Gb4 to generate the free amino sugar containing species (aminoGb4) results in the generation of a glycolipid preferentially recognized by all members of the verotoxin family (i.e., VT1, VT2, VT2c, and VT2e). GT3, a site-specific mutant of VT2e, in which Gb4 recognition is lost but Gb3 binding is retained, also binds aminoGb4. We have now compared the binding of VT1, VT2, VT2e, and GT3 to a series of aminoGb4 derivatives using a TLC overlay technique. DimethylaminoGb4 is bound by VT1 and VT2 but not VT2e or GT3; formylaminoGb4 binds all toxins but poorly to VT2 and preferentially VT2e; trifluoroacetylaminoGb4 binds only VT2e and GT3; isopropylaminoGb4 binds VT1 and poorly to VT2; benzylaminoGb4 binds all four toxins. Thus, there is a marked distinction between the permissible amino substitutions for VT1 and VT2e binding. GT3 is a hybrid between these in that, according to the substitution, it behaves similarly either to VT1 or to VT2e. For each species, GT3 does not however, show a hybrid binding between that of VT1 and VT2e. Analysis of the binding as a function of pH shows opposite effects for VT1 and VT2e: decreased pH increases VT1, but decreases VT2e receptor glycolipid binding.

Description: This paper presents the results of two field studies to examine the differences in ambient temperatures in a micro-scale environment (at distances of 50–200 m between measuring points) in two different climates during typical summer weather conditions at two similar sites in terms of construction and activities. The analysis considered the land use around the measuring locations split into three categories (built, green and open) as well as climatic conditions and studied the effect of these on ambient temperature at each measuring location. It was found that, similarly to macro-scale studies at the urban level, measuring locations with a higher green cover have a lower ambient temperature compared with measuring locations with a higher built and/or open land cover. The results provide measured evidence in two different climates that small green areas distributed within the urban environment can provide a reduction in the ambient temperature thus contributing to the mitigation of urban heat island.

Description: Capillary pump loop (CPL) is a two-phase heat transfer device considered as a useful solution for thermal control applications in spacecrafts, satellites and electronic components. The purpose of this paper is to study various aspects of the working state of the CPL evaporator. A two-dimensional computational model was developed in order to analyze the flow and the phase transformation inside a cylindrical evaporator. At the present analysis, different cases were studied by changing liquid saturation temperature, inclination angle of the evaporator and the length of the porous heated wick. Water, ammonia, acetone and freon-134 were used as working liquids for numerical experiments. Results present the evaporator capability to vaporize each working liquid and find out its possibility of dry-out causing failure. This information is useful in choosing the best working liquid for an evaporator, enabling bigger amounts of heat to be transferred.

Description: In recent years, the concept of green building is becoming popular in China. Architects and engineers have the opportunity to actually implement green building practices. This paper reveals the failure of a naturally ventilated LEED-certified townhouse development project in China mentioned in recent research. The house owners made great alterations for the purpose of decoration, which removed or minimized the effectiveness of natural ventilation elements in the original design. The research shows that the houses show ‘International Style’ because the natural ventilation design principles were not fully applied in the design and the localized natural ventilation strategies were not fully considered by means of computational simulation in the design stage. Also, the lack of communication between designers and the owners caused the removal of most natural ventilation elements of these houses during the occupancy period. The authors advocate three criteria to avoid failures of natural ventilation design: localized/ climate adaptive design, relationship among design elements and design intention awareness.

Description: In the present paper, design of solar chimney (SC) and earth-to-air heat exchanger (EAHE) to meet the thermal need of flat buildings are studied regarding adaptive thermal comfort criteria. Investigation on the effects of geometric features shows that the design of SC with the air gap and outlet sizes of 0.2 m and also EAHE with the diameter and length of 0.5 and 25.0 m reveals better performance. Thermal comfort analysis shows that the SC is capable to power the underground heating system during few hours of the sunny days even at the ambient temperature as low as 0°C and the heating demand of 1000 W without needing the auxiliary devices. In addition, the required numbers of SCs and the underground air channels are strongly influenced by environmental outdoor conditions and heating demand of building and are approximately calculated by: $$\hbox{ room }\phantom{\rule{0.08em}{0ex}}\hbox{ volume/50 }\phantom{\rule{0.08em}{0ex}}\hbox{ and }\phantom{\rule{0.08em}{0ex}}\hbox{ 2 }\times (\hbox{ room }\phantom{\rule{0.08em}{0ex}}\hbox{ volume/50 })+\hbox{ 1, }$$ respectively.

Description: Daylighting has an important role in sustainable architecture as it affects the energy consumption in a building. Especially for an office building, daylight helps not only to reduce the load on artificial lighting but also to promote work efficiency through promoting good human health, well-being and user comfort. The objective of this research is to develop an innovative façade design strategy that comes from the development of digital technology and dynamic daylight performance measuring methods. Thus, the various parameters are studied through the computational process of cellular automata (CA) to generate the several alternative opening patterns on the building façade. Each CA design value was tested under static and dynamic sky condition to analyze the quality and quantity of daylight and visual comfort throughout the year. The results were compared to find an optimum alternative design in terms of the daylighting design criteria, from building code, standard and design guidelines for the office building. Finally, research of an adaptive façade design strategy was concluded with the results from the above hybridization of generative and performative design methodology. This study discovers the architectural design approach from the CA and it will make not only progress in building façade esthetics, but also human comfort with building sustainability.

Description: It is well known that the Low-Voltage DC (LVDC) distribution system is a promising topology as a future smart distribution system due to its high efficiency and reliability. However, there are still some challenges in the construction and implementation of an LVDC system. For practical application of the LVDC system, therefore, it is necessary to perform any simulation in advance by considering various conditions that can occur in an LVDC system. In order to provide a foundation for analyzing a DC system, this paper presents an LVDC distribution system model including essential components such as power electronic devices, Distributed Energy Resource (DER), and Energy Storage System (ESS), which can be considered for implementation in an LVDC system using Electro-Magnetic Transient Program (EMTP) software. Moreover, an analysis of the characteristic in both the steady state and the transient state is conducted in an LVDC distribution system.

Description: Jordan is characterized as a “water scarce” country. Therefore, conserving ecosystem services such as water regulation and soil retention is challenging. In Jordan, rainwater harvesting has been adapted to meet those challenges. However, the spatial composition and configuration features of a target landscape are rarely considered when selecting a rainwater-harvesting site. This study aimed to introduce landscape spatial features into the schemes for selecting a proper water-harvesting site. Landscape metrics analysis was used to quantify 10 metrics for three potential landscapes (i.e., Watershed 104 (WS 104), Watershed 59 (WS 59), and Watershed 108 (WS 108)) located in the Jordanian Badia region. Results of the metrics analysis showed that the three non–vegetative land cover types in the three landscapes were highly suitable for serving as rainwater harvesting sites. Furthermore, Analytic Hierarchy Process (AHP) was used to prioritize the fitness of the three target sites by comparing their landscape metrics. Results of AHP indicate that the non-vegetative land cover in the WS 104 landscape was the most suitable site for rainwater harvesting intervention, based on its dominance, connectivity, shape, and low degree of fragmentation. Our study advances the water harvesting network design by considering its landscape spatial pattern.

Description: Since the 90s, several studies were conducted to evaluate the predictability of the Sahelian rainy season and propose seasonal rainfall forecasts to help stakeholders to take the adequate decisions to adapt with the predicted situation. Unfortunately, two decades later, the forecasting skills remains low and forecasts have a limited value for decision making while the population is still suffering from rainfall interannual variability: this shows the limit of commonly used predictors and forecast approaches for this region. Thus, this paper developed and tested new predictors and new approaches to predict the upcoming seasonal rainfall amount over the Sirba watershed. Predictors selected through a linear correlation analysis were further processed using combined linear methods to identify those having high predictive power. Seasonal rainfall was forecasted using a set of linear and non-linear models. An average lag time up to eight months was obtained for all models. It is found that the combined linear methods performed better than non-linear, possibly because non-linear models require larger and better datasets for calibration. The R2, Nash and Hit rate score are respectively 0.53, 0.52, and 68% for the combined linear approach; and 0.46, 0.45, 61% for non-linear principal component analysis.

Description: Fulvic (FAs) and humic acids (HAs) are chemically fascinating. In water, they have a strong propensity to aggregate, but this research reveals that tendency is regulated by ionic strength. In the environment, conductivity extremes occur naturally—freshwater to seawater—warranting consideration at low and high values. The flow field flow fractionation (flow FFF) of FAs and HAs is observed to be concentration dependent in low ionic strength solutions whereas the corresponding flow FFF fractograms in high ionic strength solutions are concentration independent. Dynamic light scattering (DLS) also reveals insight into the conductivity-dependent behavior of humic substances (HSs). Four particle size ranges for FAs and humic acid aggregates are examined: (1) 〈10 nm; (2) 10 nm–6 µm; (3) 6–100 µm; and (4) >100 µm. Representative components of the different size ranges are observed to dynamically coexist in solution. The character of the various aggregates observed—such as random-extended-coiled macromolecules, hydrogels, supramolecular, and micellar—as influenced by electrolytic conductivity, is discussed. The disaggregation/aggregation of HSs is proposed to be a dynamic equilibrium process for which the rate of aggregate formation is controlled by the electrolytic conductivity of the solution.

Description: Global climate change is projected to adversely impact freshwater resources, and in many settings these impacts are already apparent. In Nigeria, these impacts can be especially severe because of limited adaptive capacity. Understanding the knowledge and attitudes of current and future Nigerian decision-makers is important to preparing Nigeria for climate change impacts. This paper examines the knowledge and attitudes of university students and government officials about the causes, effects, and priority given to climate change in Nigeria. Paper surveys were distributed to 379 study participants in Akwa Ibom and Lagos states of Nigeria. The findings reveal that approximately 90% of study participants believe that human activities are a significant cause of climate change, with no significant difference between ministry officials’ and students’ responses. Participants were less knowledgeable about the effects of climate change on Nigeria as a whole, but more aware of impacts relevant to Southern Nigeria, where study sites were located. Personal experience seemed to play a role in the knowledge and attitudes of respondents. Due to the varied climate of the country, campaigns to ensure comprehensive knowledge of climate change impacts to the entire country may be helpful.

Description: The demand for thermal coal has risen dramatically over the past 20 years, driven by an emerging markets-led commodities super cycle, which has recently come to an end. Today, thermal coal generates over 40 per cent of global electricity and is available in large quantities in countries such as China, India and the USA. Thermal coal is also the primary contributor to CO 2 emissions, a substantial driver of climate change. This has resulted in material tension for ‘sustainable and responsible’ institutional investors and strategic challenges for certain state-owned companies. Deciding upon the electricity generation technology in which to invest poses a significant dilemma. Thermal coal is abundant and by far the cheapest when externalities are not internalized. Electric renewables currently suffer from intermittency, viable storage solutions and are not very scalable. While cleaner than thermal coal, natural gas is abundant yet often needs to be imported and is more expensive than thermal coal. Although safe, scalable and clean from an emissions perspective, ‘new’ nuclear energy suffers from perception problems. The mixed method inductive methodology was used to determine the extent to which thermal coal remains investable over the next 20 years and has led to two divergent yet plausible scenarios impacting thermal coal investability. In assessing the investability of thermal coal across four chief stakeholders and geographies, a multifaceted interpretation of the term ‘investability’ is developed, which recognizes disparate financial and non-financial investment drivers. The research shows that thermal coal remains investable by a variety of stakeholders, particularly in the Transformative scenario, but their reasons for investing differ materially both by geography and the type of stakeholder. Finally, the key drivers for both scenarios are identified that can be monitored and used as an early warning system to inform investment decisions.

Description: In March of 2015 New Zealand’s Court of Appeal brought to a conclusion a long running dispute between the joint venture partners in the Pohokura gas and condensate field located in Taranaki, New Zealand. Todd Pohokura v. Shell Exploration NZ Limited et ano deals with the rights to production and offtake arising under the Association of International Petroleum Negotiator’s (AIPN) 1995 Model Form joint operating agreement (JOA), the powers of the Operating Committee in regards thereto and the practical implications of entering into a field development without documenting all the necessary gas sale and transportation arrangements in advance. The factual matrix provides an opportunity to review the principles of common law relevant to production and offtake, including the development of those principles from early English common law into U.S. domestic law, and how those principles have been reflected in model form JOAs on both sides of the Atlantic, including the AIPN forms and ultimately the Pohokura JOA. The analysis confirms the New Zealand courts’ rulings upholding the power of the Operating Committee to determine production rates under the AIPN Model form. However, it concludes that gas offtake arrangements that amend or add to the rights and obligations of the parties under the JOA must be agreed unanimously and are not within the jurisdiction of the Operating Committee to determine.

Description: Determining the optimum angle for a solar panel is important if tracking systems are not used and a tilt angle remains constant. This article determines the sensitivity of the optimum angle to surface reflectivity at different latitudes using a mathematical model that accounts for direct, diffuse and reflected radiation. A quadratic correlation is also developed to compute the optimal angle and maximum energy as a function of latitude and reflectivity. We also seek to determine how sensitive the optimal tilt angle is to cloud cover using the 35° latitude of the Prosperity solar facility in Albuquerque, NM.

Description: In sub-Saharan Africa, hydro-meteorological related disasters, such as floods, account for the majority of the total number of natural disasters. Over the past century, floods have affected 38 million people, claimed several lives and caused substantial economic losses in the region. The goal of this paper is to examine how personality disposition, social network, and socio-demographic factors mitigate the complex relationship between stressful life experiences of floods and ocean surges and the adoption of coping strategies among coastal communities in Nigeria and Tanzania. Generalized linear models (GLM) were fitted to cross-sectional survey data on 1003 and 1253 individuals in three contiguous coastal areas in Nigeria and Tanzania, respectively. Marked differences in the type of coping strategies were observed across the two countries. In Tanzania, the zero-order relationships between adoption of coping strategies and age, employment and income disappeared at the multivariate level. Only experience of floods in the past year and social network resources were significant predictors of participants’ adoption of coping strategies, unlike in Nigeria, where a plethora of factors such as experience of ocean surges in the past one year, personality disposition, age, education, experience of flood in the past one year, ethnicity, income, housing quality and employment status were still statistically significant at the multivariate level. Our findings suggest that influence of previous experience on adoption of coping strategies is spatially ubiquitous. Consequently, context-specific policies aimed at encouraging the adoption of flood-related coping strategies in vulnerable locations should be designed based on local needs and orientation.

Description: Ferritins and other cage proteins have been utilized as models to understand the fundamentals of protein folding and self-assembly. The bacterioferritin (BFR) from Escherichia coli, a maxi-ferritin made up of 24 subunits, was chosen as the basis for a mutagenesis study to investigate the role of electrostatic intermolecular interactions mediated through charged amino acids. Through structural and computational analyses, three charged amino acids R30, D56 and E60 which involved in an electrostatic interaction network were mutated to the opposite charge. Four mutants, R30D, D56R, E60H and D56R-E60H, were expressed, purified and characterized. All of the mutants fold into α-helical structures. Consistent with the computational prediction, they all show a lowered thermostability; double mutant D56R-E60H was found to be 16°C less stable than the wild type. Except for the mutant E60H, all the other mutations completely shut down the formation of protein cages to favour the dimer state in solution. The mutants, however, retain their ability to form cage-like nanostructures in the dried, surface immobilized conditions of transmission electron microscopy. Our findings confirm that even a single charge-inversion mutation at the 2-fold interface of BFR can affect the quaternary structure of its dimers and their ability to self-assemble into cage structures.

Description: Most of bacteria can swim by rotating flagella bidirectionally. The C ring, located at the bottom of the flagellum and in the cytoplasmic space, consists of FliG, FliM and FliN, and has an important function in flagellar protein secretion, torque generation and rotational switch of the motor. FliG is the most important part of the C ring that interacts directly with a stator subunit. Here, we introduced a three-amino acids in-frame deletion mutation (PSA) into FliG from Vibrio alginolyticus , whose corresponding mutation in Salmonella confers a switch-locked phenotype, and examined its phenotype. We found that this FliG mutant could not produce flagellar filaments in a fliG null strain but the FliG(PSA) protein could localize at the cell pole as does the wild-type protein. Unexpectedly, when this mutant was expressed in a wild-type strain, cells formed flagella efficiently but the motor could not rotate. We propose that this different phenotype in Vibrio and Salmonella might be due to distinct interactions between FliG mutant and FliM in the C ring between the bacterial species.

Description: Sulphation is known to be critically involved in the metabolism of acetaminophen in vivo . This study aimed to systematically identify the major human cytosolic sulfotransferase (SULT) enzyme(s) responsible for the sulphation of acetaminophen. A systematic analysis showed that three of the twelve human SULTs, SULT1A1, SULT1A3 and SULT1C4, displayed the strongest sulphating activity towards acetaminophen. The pH dependence of the sulphation of acetaminophen by each of these three SULTs was examined. Kinetic parameters of these three SULTs in catalysing acetaminophen sulphation were determined. Moreover, sulphation of acetaminophen was shown to occur in HepG2 human hepatoma cells and Caco-2 human intestinal epithelial cells under the metabolic setting. Of the four human organ samples tested, liver and intestine cytosols displayed considerably higher acetaminophen-sulphating activity than those of lung and kidney. Collectively, these results provided useful information concerning the biochemical basis underlying the metabolism of acetaminophen in vivo previously reported.

Description: In this study, the physicochemical and enzymatic properties of recombinant human ubiquitin (Ub)-specific protease (USP) 47, a novel member of the C19 family of de-ubiquitinating enzymes (DUB), were characterized for the first time. Recombinant human USP47 was expressed in a baculovirus expression system and purified to homogeneity. The purified protein was shown to be a monomeric protein with a molecular mass of ~146 kDa on sodium dodecyl sulphate—polyacrylamide gel electrophoresis. USP47 released Ub from Ub-aminoacyl-4-metheylcoumaryl-7-amide and Ub-tagged granzyme B. The substitution of the potential nucleophile Cys109 with Ser severely abrogated the Ub-releasing activity of USP47, indicating that USP47 is indeed a cysteine DUB. An assay using Ub dimer substrates showed that the enzyme cleaved a variety of isopeptide bonds between 2 Ub molecules, including the Lys48- and Lys63-linked isopeptide bonds. USP47 also released a Ub moiety from Lys48- and Lys63-linked polyUb chains. Of the inhibitors tested, N -ethylmaleimide, Zn ion and Ub aldehyde revealed a dose-dependent inhibition of USP47. In this study, clear differences in the enzymatic properties between USP47 and USP7 (the most closely related proteins among DUBs) were also found. Therefore, our results suggest that USP47 may play distinct roles in Ub-mediated cellular processes via DUB activity.

Description: P24 antigen is the main structural protein of HIV-1, its detection provide a means to aid the early diagnosis of HIV-1 infection. The aim of this study was to improve the selectivity and sensitivity of the HIV P24 diagnostic assay by developing a cohort of 9E8 affinity-matured antibodies through in vitro phage affinity maturation which was performed by complementarity determining region (CDR)-hot spot mutagenesis strategy. Antibody 9E8-491 had an affinity constant of 5.64 x 10 –11 M, which was 5.7-fold higher than that of the parent antibody (9E8). Furthermore, the affinity, sensitivity and specificity of 9E8-491 were higher than those of 9E8, which indicate that 9E8-491 is a good candidate detection antibody for HIV P24 assay. Structure analysis of matured variants revealed that most hydrogen bonds resided in HCDR3. Among the antibody–antigen predicted binding residues, Tyr 100A/100B was the original conserved residue that was commonly present in HCDR3 of 9E8 and variants. Arg 100 /Asp 100C was the major variant substitution that most likely influenced the binding differences among variants and 9E8 monoclonal antibody. Both efficient library panning and predicted structural data were in agreement that the binding residues were mostly located in HCDR3 and enabled identification of key residues that influence antibody affinity.

Description: Climate change impacts on nature and the environment have been widely discussed and studied. Traditionally, a company’s continuity management is based on risk analysis. There are also attempts to implement scenario-based methods in the risk management procedures of companies. For industrial decision makers, it is vital to acknowledge the impacts of climate change with regards to their adaptation strategies. However, a scenario-based approach is not always the most effective way to analyze these risks. This paper investigates the integration of scenario and risk-based methods for a company’s adaptation planning. It considers the uncertainties of the climate change scenarios and the recognized risks as well as suitable adaptation strategies. The paper presents the results of climate risk analysis prepared for two Finnish hydropower plants. The introduced method was first piloted in 2008 and then again in 2015. The update of the analysis pointed out that at the company level, the climate risks and other risks originating from governmental or political decisions form an intertwined wholeness where the origin of the risk is difficult to outline. It seems that, from the business point of view, the main adaptation strategies suggested by the integrated risk and scenarios approach are those that support buying “safety margins” in new investments and reducing decision time horizons. Both of these adaptation strategies provide an advantage in the circumstances where also political decisions and societal changes have a great effect on decision making.

Description: Many skeletal diseases have common pathological phenotype of defective osteogenesis of bone marrow stromal cells (BMSCs), in which histone modifications play an important role. However, few studies have examined the dynamics of distinct histone modifications during osteogenesis. In this study, we examined the dynamics of H3K9/K14 and H4K12 acetylation; H3K4 mono-, di- and tri-methylation; H3K9 di-methylation and H3K27 tri-methylation in osteogenic genes, runt-related transcription factor 2 (Runx2), osterix (Osx), alkaline phosphatase, bone sialoprotein and osteocalcin, during C3H10T1/2 osteogenesis. H3 and H4 acetylation and H3K4 di-methylation were elevated, and H3K9 di-methylation and H3K27 tri-methylation were reduced in osteogenic genes during C3H10T1/2 osteogenesis. C3H10T1/2 osteogenesis could be modulated by altering the patterns of H3 and H4 acetylation and H3K27 tri-methylation. In a glucocorticoid-induced osteoporosis mouse model, we observed the attenuation of osteogenic potential of osteoporotic BMSCs in parallel with H3 and H4 hypo-acetylation and H3K27 hyper-tri-methylation in Runx2 and Osx genes. When H3 and H4 acetylation was elevated, and H3K27 tri-methylation was reduced, the attenuated osteogenic potential of osteoporotic BMSCs was rescued effectively. These observations provide a deeper insight into the mechanisms of osteogenic differentiation and the pathophysiology of osteoporosis and can be used to design new drugs and develop new therapeutic methods to treat skeletal diseases.

Description: Dihydrouridine (D) is formed by tRNA dihydrouridine synthases (Dus). In mesophiles, multiple Dus enzymes bring about D modifications at several positions in tRNA. The extreme-thermophilic eubacterium Thermus thermophilus , in contrast, has only one dus gene in its genome and only two D modifications (D20 and D20a) in tRNA have been identified. Until now, an in vitro assay system for eubacterial Dus has not been reported. In this study, therefore, we constructed an in vitro assay system using purified Dus. Recombinant T. thermophilus Dus lacking bound tRNA was successfully purified. The in vitro assay revealed that no other factors in living cells were required for D formation. A dus gene disruptant ( dus ) strain of T. thermophilus verified that the two D20 and D20a modifications in tRNA were derived from one Dus protein. The dus strain did not show growth retardation at any temperature. The assay system showed that Dus modified tRNA Phe transcript at 60°C, demonstrating that other modifications in tRNA are not essential for Dus activity. However, a comparison of the formation of D in native tRNA Phe purified from the dus strain and tRNA Phe transcript revealed that other tRNA modifications are required for D formation at high temperatures.

Description: Human lactate dehydrogenase (LDH) has attracted attention as a potential target for cancer therapy and contraception. In this study, we reconstituted human lactic acid fermentation in Saccharomyces cerevisiae , with the goal of constructing a yeast cell-based LDH assay system. pdc null mutant yeast (mutated in the endogenous pyruvate decarboxylase genes) are unable to perform alcoholic fermentation; when grown in the presence of an electron transport chain inhibitor, pdc null strains exhibit a growth defect. We found that introduction of the human gene encoding LDHA complemented the pdc growth defect; this complementation depended on LDHA catalytic activity. Similarly, introduction of the human LDHC complemented the pdc growth defect, even though LDHC did not generate lactate at the levels seen with LDHA. In contrast, the human LDHB did not complement the yeast pdc null mutant, although LDHB did generate lactate in yeast cells. Expression of LDHB as a red fluorescent protein (RFP) fusion yielded blebs in yeast, whereas LDHA-RFP and LDHC-RFP fusion proteins exhibited cytosolic distribution. Thus, LDHB exhibits several unique features when expressed in yeast cells. Because yeast cells are amenable to genetic analysis and cell-based high-throughput screening, our pdc /LDH strains are expected to be of use for versatile analyses of human LDH.

Description: RelB is activated by the non-canonical NF-B pathway, which is crucial for immunity by establishing lymphoid organogenesis and B-cell and dendritic cell (DC) maturation. To elucidate the mechanism of the RelB-mediated immune cell maturation, a precise understanding of the relationship between cell maturation and RelB expression and activation at the single-cell level is required. Therefore, we generated knock-in mice expressing a fusion protein between RelB and fluorescent protein (RelB-Venus) from the Relb locus. The Relb Venus / Venus mice developed without any abnormalities observed in the Relb –/– mice, allowing us to monitor RelB-Venus expression and nuclear localization as RelB expression and activation. Relb Venus / Venus DC analyses revealed that DCs consist of RelB – , RelB low and RelB high populations. The RelB high population, which included mature DCs with projections, displayed RelB nuclear localization, whereas RelB in the RelB low population was in the cytoplasm. Although both the RelB low and RelB – populations barely showed projections, MHC II and co-stimulatory molecule expression were higher in the RelB low than in the RelB – splenic conventional DCs. Taken together, our results identify the RelB low population as a possible novel intermediate maturation stage of cDCs and the Relb Venus / Venus mice as a useful tool to analyse the dynamic regulation of the non-canonical NF-B pathway.

Description: Hyperthermophilic bacteria Thermotoga maritima and Thermotoga hypogea produce ethanol as a metabolic end product, which is resulted from acetaldehyde reduction catalysed by an alcohol dehydrogenase (ADH). However, the enzyme that is involved in the production of acetaldehyde from pyruvate is not well characterized. An oxygen sensitive and coenzyme A-dependent pyruvate decarboxylase (PDC) activity was found to be present in cell free extracts of T. maritima and T. hypogea . Both enzymes were purified and found to have pyruvate ferredoxin oxidoreductase (POR) activity, indicating their bifunctionality. Both PDC and POR activities from each of the purified enzymes were characterized in regards to their optimal assay conditions including pH dependency, oxygen sensitivity, thermal stability, temperature dependency and kinetic parameters. The close relatedness of the PORs that was shown by sequence analysis could be an indication of the presence of such bifunctionality in other hyperthermophilic bacteria. This is the first report of a bifunctional PDC/POR enzyme in hyperthermophilic bacteria. The PDC and the previously reported ADHs are most likely the key enzymes catalysing the production of ethanol from pyruvate in bacterial hyperthermophiles.

Description: Energy efficiency has been an important topic since the latter part of the last century. This is because adoption of energy efficiency measures has been acknowledged as one of the key methods of addressing the negative impact of climate change. In Zambia, however, the need to adopt energy efficiency measures has not just been driven by the imperative to mitigate the negative effects of climate change but also by a critical shortage of energy. This research looks at households’ energy consumption behavior in low- and high-income areas of Kitwe. Recent studies on the relationship between household energy consumption and behavioral lifestyle have been descriptive, with limited emphasis on the relationships between various variables. In this study, descriptive and inferential statistics have been used to investigate relationships between the two income groups and various energy consumption-related variables such as knowledge about energy reduction measures, energy saving strategies, barriers to the use of energy saving strategies, and the motives for using energy reduction strategies. Methodologically, the study was largely quantitative in nature, with questionnaires administered to a combined total of 56 households. However, key interviews were also conducted that helped us to get a clearer understanding of some of the issues covered in the research. Key findings are that whereas the descriptive statistics show that there are behavioral differences between the two income groups, the inferential statistics show that there is no relationship between income level and the energy efficiency variables. This has been found to be consistent with results from studies done elsewhere. The key lesson is that there is low usage of energy efficiency measures in both low- and high-income areas and that the authorities need to change the way information is disseminated to consumers from the current method of advertising to social diffusion.

Description: Magnetic nanoparticles (MNPs) exhibit unique magnetic properties making them ideally suited for a variety of biomedical applications. Depending on the desired magnetic effect, MNPs must meet special magnetic requirements which are mainly determined by their structural properties (e.g., size distribution). The hyphenation of chromatographic separation techniques with complementary detectors is capable of providing multidimensional information of submicron particles. Although various methods have already been combined for this approach, so far, no detector for the online magnetic analysis was used. Magnetic particle spectroscopy (MPS) has been proven a straightforward technique for specific quantification and characterization of MNPs. It combines high sensitivity with high temporal resolution; both of these are prerequisites for a successful hyphenation with chromatographic separation. We demonstrate the capability of MPS to specifically detect and characterize MNPs under usually applied asymmetric flow field-flow fractionation (A4F) conditions (flow rates, MNP concentration, different MNP types). To this end MPS has been successfully integrated into an A4F multidetector platform including dynamic ligth scattering (DLS), multi-angle light scattering (MALS) and ultraviolet (UV) detection. Our system allows for rapid and comprehensive characterization of typical MNP samples for the systematic investigation of structure-dependent magnetic properties. This has been demonstrated by magnetic analysis of the commercial magnetic resonance imaging (MRI) contrast agent Ferucarbotran (FER) during hydrodynamic A4F fractionation.

Description: The scaffolding protein Salvador (Sav) plays a key role in the Hippo (Hpo) signalling pathway, which controls tissue growth by inhibiting cell proliferation and promoting apoptosis. Dysregulation of the Hippo pathway contributes to cancer development. Since the identification of the first Sav gene in 2002, very little is known regarding the molecular basis of Sav-SARAH mediating interactions due to its insolubility. In this study, refolding of the first Sav (known as WW45)-SARAH provided insight into the biochemical and biophysical properties, indicating that WW45-SARAH exhibits properties of a disordered protein, when the domain was refolded at a neutral pH. Interestingly, WW45-SARAH shows folded and rigid conformations relative to the decrease in pH. Further, diffracting crystals were obtained from protein refolded under acidic pH, suggesting that the refolded WW45 protein at low pH has a homogeneous and stable conformation. A comparative analysis of molecular properties found that the acidic-stable fold of WW45-SARAH enhances a heterotypic interaction with Mst2-SARAH. In addition, using an Mst2 mutation that disrupts homotypic dimerization, we showed that the monomeric Mst2-SARAH domain could form a stable complex of 1:1 stoichiometric ratio with WW45 refolded under acidic pH.

Description: Hypercholesterolemia is one of the factors contributing to cardiovascular problems. Erythrocytes are known to contribute its cholesterol to atherosclerotic plaque. Our earlier study showed that erythrocytes overexpress chondroitin sulphate/dermatan sulphate (CS/DS), a linear co-polymer, during diabetes which resulted in increased cytoadherence to extracellular matrix (ECM) components. This study was carried out to determine whether diet-induced hypercholesterolemia had any effect on erythrocyte CS/DS and impacted cytoadherence to ECM components. Unlike in diabetes, diet-induced hypercholesterolemia did not show quantitative changes in erythrocyte CS/DS but showed difference in proportion of un-sulphated and 4- O -sulphated disaccharides. Erythrocytes from hypercholesterolemic rats showed increased adhesion to ECM components which was abrogated to various extents when subjected to chondroitinase ABC digestion. However, isolated CS/DS chains showed a different pattern of binding to ECM components indicating that orientation of CS/DS chains could be playing a role in binding.

Description: The antigen-binding domain of camelid dimeric heavy chain antibodies, known as VHH or Nanobody, has much potential in pharmaceutical and industrial applications. To establish the isolation process of antigen-specific VHH, a VHH phage library was constructed with a diversity of 8.4 x 10 7 from cDNA of peripheral blood mononuclear cells of an alpaca ( Lama pacos ) immunized with a fragment of IZUMO1 (IZUMO1 PFF ) as a model antigen. By conventional biopanning, 13 antigen-specific VHHs were isolated. The amino acid sequences of these VHHs, designated as N-group VHHs, were very similar to each other (〉93% identity). To find more diverse antibodies, we performed high-throughput sequencing (HTS) of VHH genes. By comparing the frequencies of each sequence between before and after biopanning, we found the sequences whose frequencies were increased by biopanning. The top 100 sequences of them were supplied for phylogenic tree analysis. In total 75% of them belonged to N-group VHHs, but the other were phylogenically apart from N-group VHHs (Non N-group). Two of three VHHs selected from non N-group VHHs showed sufficient antigen binding ability. These results suggested that biopanning followed by HTS provided a useful method for finding minor and diverse antigen-specific clones that could not be identified by conventional biopanning.

Description: The autophosphorylation of specific tyrosine residues occurs in the cytoplasmic region of the insulin receptor (IR) upon insulin binding, and this in turn initiates signal transduction. The R3 subfamily (Ptprb, Ptprh, Ptprj and Ptpro) of receptor-like protein tyrosine phosphatases (RPTPs) is characterized by an extracellular region with 6–17 fibronectin type III-like repeats and a cytoplasmic region with a single phosphatase domain. We herein identified the IR as a substrate for R3 RPTPs by using the substrate-trapping mutants of R3 RPTPs. The co-expression of R3 RPTPs with the IR in HEK293T cells suppressed insulin-induced tyrosine phosphorylation of the IR. In vitro assays using synthetic phosphopeptides revealed that R3 RPTPs preferentially dephosphorylated a particular phosphorylation site of the IR: Y960 in the juxtamembrane region and Y1146 in the activation loop. Among four R3 members, only Ptprj was co-expressed with the IR in major insulin target tissues, such as the skeletal muscle, liver and adipose tissue. Importantly, the activation of IR and Akt by insulin was enhanced, and glucose and insulin tolerance was improved in Ptprj -deficient mice. These results demonstrated Ptprj as a physiological enzyme that attenuates insulin signalling in vivo , and indicate that an inhibitor of Ptprj may be an insulin-sensitizing agent.

Description: The diazotrophic cyanobacterium Anabaena sp. strain PCC 7120 (A.7120) differentiates into specialized heterocyst cells that fix nitrogen under nitrogen starvation conditions. Although reducing equivalents are essential for nitrogen fixation, little is known about redox systems in heterocyst cells. In this study, we investigated thioredoxin (Trx) networks in Anabaena using TrxM, and identified 16 and 38 candidate target proteins in heterocysts and vegetative cells, respectively, by Trx affinity chromatography (Motohashi et al. (Comprehensive survey of proteins targeted by chloroplast thioredoxin. Proc Natl Acad Sci USA , 2001; 98 , 11224–11229)). Among these, the Fe–S cluster scaffold protein NifU that facilitates functional expression of nitrogenase in heterocysts was found to be a potential TrxM target. Subsequently, we observed that the scaffold activity of N-terminal catalytic domain of NifU is enhanced in the presence of Trx-system, suggesting that TrxM is involved in the Fe–S cluster biogenesis.

Description: A detailed statistical analysis was performed at the Neuquén river basin using precipitation data for 1980–2007. The hydrological year begins in March with a maximum in June associated with rainfall and another relative maximum in October derived from snow-break. General features of the rainy season and the excess or deficits thereof are analyzed using standardized precipitation index (SPI) for a six-month period in the basin. The SPI has a significant cycle of 14.3 years; the most severe excess (SPI greater than 2) has a return period of 25 years, while the most severe droughts (SPI less than −2) have a return period of 10 years. The SPI corresponding to the rainy season (April–September) (SPI9) has no significant trend and is used to classify wet/dry years. In order to establish the previous circulation patterns associated with interannual SPI9 variability, the composite fields of wet and dry years are compared. There is a tendency for wet (dry) periods to take place during El Niño (La Niña) years, when there are positive anomalies of precipitable water over the basin, when the zonal flow over the Pacific Ocean is weakened (intensified) and/or when there are negative pressure anomalies in the southern part of the country and Antarctic sea. Some prediction schemes using multiple linear regressions were performed. One of the models derived using the forward stepwise method explained 42% of the SPI9 variance and retained two predictors related to circulation over the Pacific Ocean: one of them shows the relevance of the intensity of zonal flow in mid-latitudes, and the other is because of the influence of low pressure near the Neuquén River basin. The cross-validation used to prove model efficiency showed a correlation of 0.41 between observed and estimated SPI9; there was a probability of detection of wet (dry) years of 80% (65%) and a false alarm relation of 25% in both cases.

Description: The Abdus Salam International Center for Theoretical Physics (ICTP) version 4.4 Regional Climate Model (RegCM4) is used to investigate the rainfall response to cooler/warmer sea surface temperature anomaly (SSTA) forcing in the Indian and Atlantic Oceans. The effect of SSTA forcing in a specific ocean basin is identified by ensemble, averaging 10 individual simulations in which a constant or linearly zonally varying SSTA is prescribed in individual basins while specifying the 1971–2000 monthly varying climatological sea surface temperature (SST) across the remaining model domain. The nonlinear rainfall response to SSTA amplitude also is investigated by separately specifying +1K, +2K, and +4K SSTA forcing in the Atlantic and Indian Oceans. The simulation results show that warm SSTs over the entire Indian Ocean produce drier conditions across the larger Blue Nile catchment, whereas warming ≥ +2K generates large positive rainfall anomalies exceeding 10 mm·day−1 over drought prone regions of Northeastern Ethiopia. However, the June–September rainy season tends to be wetter (drier) when the SST warming (cooling) is limited to either the Northern or Southern Indian Ocean. Wet rainy seasons generally are characterized by deepening of the monsoon trough, east of 40°E, intensification of the Mascarene high, strengthening of the Somali low level jet and the tropical easterly jet, enhanced zonal and meridional vertically integrated moisture fluxes, and steeply vertically decreasing moist static energy. The opposite conditions hold for dry monsoon seasons.

Description: Pursuant to a generous Association of International Petroleum Negotiator (AIPN) 2014 Summer Research Award, this article identifies, evaluates and compares the legal and fiscal rules, regulations and incentives necessary for countries with significant shale petroleum and natural gas formations to attempt to replicate the boom that is ongoing in the USA. As others have pointed out, 1 several legal, tax, and operational barriers can impair duplication of the US shale revolution in similarly endowed nations. This article identifies key factors responsible for the surge in US shale production, distill the fundamental forces from the US experience that are applicable to any jurisdiction, and evaluate and compare how several countries fare in this vein. The report also identifies avenues for reform and innovative policies that could be applied in other jurisdictions.

Description: Texas law has only recently codified in precedential decisions a reasonable formula for production allocations for horizontal, including fracked, wells. The ancient regime of the Rule of Capture does not generally apply, because of the nature of the geologic deposits and the new technology. This reasonable formula is applicable in the international arena for cross-border deposits and production blocks. Other national law and international treaties provide scant guidance in these matters and governments, national oil companies and international oil companies should review this formula to aid in negotiations for fair and equitable allocations that should preclude unnecessary disputes and litigation.

Description: A new dynamical downscaling methodology to analyze the impact of global climate change on the local climate of cities worldwide is presented. The urban boundary layer climate model UrbClim is coupled to 11 global climate models contained in the Coupled Model Intercomparison Project 5 archive, conducting 20-year simulations for present (1986–2005) and future (2081–2100) climate conditions, considering the Representative Concentration Pathway 8.5 climate scenario. The evolution of the urban heat island of eight different cities, located on three continents, is quantified and assessed, with an unprecedented horizontal resolution of a few hundred meters. For all cities, urban and rural air temperatures are found to increase strongly, up to 7 °C. However, the urban heat island intensity in most cases increases only slightly, often even below the range of uncertainty. A potential explanation, focusing on the role of increased incoming longwave radiation, is put forth. Finally, an alternative method for generating urban climate projections is proposed, combining the ensemble temperature change statistics and the results of the present-day urban climate.

Description: In this study, we examined the role of aminopeptidases with reference to endoplasmic reticulum aminopeptidase 1 (ERAP1) in nitric oxide (NO) synthesis employing murine macrophage cell line RAW264.7 cells activated by lipopolysaccharide (LPS) and interferon (IFN)- and LPS-activated peritoneal macrophages derived from ERAP1 knockout mouse. When NO synthesis was measured in the presence of peptides having N-terminal Arg, comparative NO synthesis was seen with that measured in the presence of Arg. In the presence of an aminopeptidase inhibitor amastatin, NO synthesis in activated RAW264.7 cells was significantly decreased. These results suggest that aminopeptidases are involved in the NO synthesis in activated RAW264.7 cells. Subsequently, significant reduction of NO synthesis was observed in ERAP1 knockdown cells compared with wild-type cells. This reduction was rescued by exogenously added ERAP1. Furthermore, when peritoneal macrophages prepared from ERAP1 knockout mouse were employed, reduction of NO synthesis in knockout mouse macrophages was also attributable to ERAP1. In the presence of amastatin, further reduction was observed in knockout mouse-derived macrophages. Taken together, these results suggest that several aminopeptidases play important roles in the maximum synthesis of NO in activated macrophages in a substrate peptide-dependent manner and ERAP1 is one of the aminopeptidases involved in the NO synthesis.

Description: Glycogen phosphorylase (GP) is biologically active as a dimer of identical subunits. Each subunit has two distinct maltooligosaccharide binding sites: a storage site and a catalytic site. Our characterization of the properties of these sites suggested that GP activity consists of two activities: (i) binding to the glycogen molecule and (ii) phosphorolysis of the non-reducing-end glucose residues. Activity (i) is mainly due to the activities of the two storage sites, which depended on the ionic strength of the medium and were directly inhibited by cyclodextrins (CDs). Activity (i) is of benefit to GP because a high concentration of non-reducing-end glucose residues is localized on the surface of the glycogen molecule. Activity (ii), the total activity of the two catalytic sites, exhibited relatively little ionic strength dependence. Because the combined activity of (i) and (ii) is deduced using glycogen as an assay substrate, the sole activity of (ii) must be measured using small maltooligosyl-substrates. By using a very low concentration of pyridylaminated maltohexaose, we demonstrated that the GP catalytic sites are active even in the presence of CDs, and that the actions of the catalytic site and the storage site are independent of each other.

Description: O -GlcNAcylation is a ubiquitous, dynamic and reversible post-translational protein modification in metazoans, and it is catalysed and removed by O -GlcNAc transferase (OGT) and O -GlcNAcase, respectively. Prokaryotes lack endogenous OGT activity. It has been reported that coexpression of mammalian OGT with its target substrates in Escherichia coli produce O -GlcNAcylated recombinant proteins, but the plasmids used were not compatible, and the expression of both OGT and its target protein were induced by the same inducer. Here, we describe a compatible dual plasmid system for coexpression of OGT and its target substrate for O -GlcNAcylated protein production in E. coli . The approach was validated using the CKII and p53 protein as control. This compatible dual plasmid system contains an arabinose-inducible OGT expression vector with a pUC origin and an isopropyl β - d -thiogalactopyranoside-inducible OGT target substrate expression vector bearing a p15A origin. The dual plasmid system produces recombinant proteins with varying O -GlcNAcylation levels by altering the inducer concentration. More importantly, the O -GlcNAcylation efficiency was much higher than the previously reported system. Altogether, we established an adjustable compatible dual plasmid system that can effectively yield O -GlcNAcylated proteins in E. coli .

Description: Active equi-paritioning of the F plasmid is achieved by its sopABC gene. SopA binds to the sopAB promoter region and SopB binds to sopC . SopA also polymerizes in the presence of ATP and Mg(II), which is stimulated by SopB. Non-specific DNA is known to inhibit SopA polymerization and disassemble SopA polymer. This study followed kinetics of polymerization and de-polymerization of SopA by turbidity measurement and found new effects by DNA and SopB. Plasmid DNA, at low concentrations, shortened the lag (nucleation) phase of SopA polymerization and also caused an initial ‘burst’ of turbidity. Results with two non-specific 20-bp DNAs indicated sequence/length dependence of these effects. sopAB operator DNA only showed inhibition of SopA polymerization. Results of turbidity decrease of pre-formed SopA polymer in the presence of ethylenediaminetetraacetic acid showed that SopB also accelerates disassembly of the SopA polymer. The steady-state level of turbidity in the presence of SopB and plasmid DNA indicated synergy between SopB and DNA in the disassembly. SopB protein showed no effect on SopA polymerization, when SopB was specifically bound to DNA. This result and others with truncation mutants of SopB suggested that a proper configuration of the domains of SopB is important for SopA-SopB interactions.

Description: Influenza A virus (IAV) has been raising public health and safety concerns worldwide. Cyanovirin-N (CVN) is a prominent anti-IAV candidate, but both cytotoxicity and immunogenicity have hindered the development of this protein as a viable therapy. In this article, linker-CVN (LCVN) with a flexible and hydrophilic polypeptide at the N-terminus was efficiently produced from the cytoplasm of Escherichia coli at a 〉15-l scale. PEGylation at the N-terminal α-amine of LCVN was also reformed as 20 kDa PEGylated linkered Cyanovirin-N (PEG 20k –LCVN). The 50% effective concentrations of PEG 20k –LCVN were 0.43 ± 0.11 µM for influenza A/HK/8/68 (H3N2) and 0.04 ± 0.02 µM for A/Swan/Hokkaido/51/96 (H5N3), dramatically lower than that of the positive control, Ribavirin (2.88 ± 0.66 x 10 3 µM and 1.79 ± 0.62 x 10 3 µM, respectively). A total of 12.5 µM PEG 20k –LCVN effectively inactivate the propagation of H3N2 in chicken embryos. About 2.0 mg/kg/day PEG 20k –LCVN increased double the survival rate (66.67%, P = 0.0378) of H3N2 infected mice, prolonged the median survival period, downregulated the mRNA level of viral nuclear protein and decreased (attenuated) the pathology lesion in mice lung. A novel PEGylated CVN derivative, PEG 20k –LCVN, exhibited potent and strain-dependent anti-IAV activity in nanomolar concentrations in vitro, as well as in micromolar concentration in vivo .

Description: The semi-filamentous multicellular cyanobacterium Limnothrix / Pseudanabaena sp. strain ABRG5-3 undergoes autolysis, which involves the accumulation of polyphosphate compounds and disintegration of thylakoid membranes in cells, as a unique feature that occurs due to growth conditions. In this study, the overexpression and easy recovery of alkane (a saturated hydrocarbon, C 17 H 36 ) as a biofuel were examined in recombinants of the cyanobacteria ABRG5-3 and Synechocystis sp. strain PCC6803. The results obtained indicated that the accumulated mass of alkane accounted for ~50 or 60% of the dry weight of ABRG5-3 or PCC6803 recombinant cells, respectively. Furthermore, cultivating cells in liquid medium BG11 in which the nitrogen resource had been depleted promoted the production of alkane and cell lysis, resulting in the easy recovery of target products from the supernatant.

Description: Leucine-rich repeat kinase 2 (LRRK2) has been identified as a causative gene for Parkinson’s disease (PD). LRRK2 contains a kinase and a GTPase domain, both of which provide critical intracellular signal-transduction functions. We showed previously that Rab5b, a small GTPase protein that regulates the motility and fusion of early endosomes, interacts with LRRK2 and co-regulates synaptic vesicle endocytosis. Using recombinant proteins, we show here that LRRK2 phosphorylates Rab5b at its Thr6 residue in in vitro kinase assays with mass spectrophotometry analysis. Phosphorylation of Rab5b by LRRK2 on the threonine residue was confirmed by western analysis using cells stably expressing LRRK2 G2019S. The phosphomimetic T6D mutant exhibited stronger GTPase activity than that of the wild-type Rab5b. In addition, phosphorylation of Rab5b by LRRK2 also exhibited GTPase activity stronger than that of the unphosphorylated Rab5b protein. Two assays testing Rab5’s activity, neurite outgrowth analysis and epidermal growth factor receptor degradation assays, showed that Rab5b T6D exhibited phenotypes that were expected to be observed in the inactive Rab5b, including longer neurite length and less degradation of EGFR. These results suggest that LRRK2 kinase activity functions as a Rab5b GTPase activating protein and thus, negatively regulates Rab5b signalling.

Description: Kratom is a tree planted in Southeast Asia, including Thailand, Malaysia, Myanmar (Burma) and elsewhere in the region. A long history of usage and abuse of kratom has led to the classification of kratom as a controlled substance in its native Thailand and other Southeast Asian countries. However, kratom is not controlled in the United States, and the wide availability of kratom on the Internet and in the streets has led to its emergence as an herbal drug of misuse. With the increasing popularity of kratom, efficient protocols are needed to detect kratom use. In this study, a rapid method for the analysis of kratom compounds, mitragynine and 7-hydroxymitragynine, in human urine has been developed and validated using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The chromatographic system employed a 2.6-μm 100 mm × 2.1 mm phenyl-hexyl analytical column and gradient elution with a 0.4-mL/min flow rate of water and acetonitrile as mobile phases. A triple quadrupole mass spectrometer was used as the detector for data acquisition. The analyst was the quantification software. The established method demonstrated linearity of >0.99 for both analytes, and low detection limits were obtained down to 0.002581 ng/mL for mitragynine and 0.06910 ng/mL for 7-hydroxymitragynine. The validated method has been utilized for clinical analysis of urine for the purpose of mitragynine and 7-hydroxymitragynine detection.

Description: The cellular Src (c-Src) tyrosine kinase is upregulated and believed to play a pivotal role in various human cancers. However, the molecular mechanism underlying c-Src-mediated tumour progression remains elusive. Recent studies have revealed that several microRNAs (miRNAs) function as tumour suppressors by regulating the malignant expression of signalling molecules. Aberrant expression of miRNAs is frequently observed in human cancers and should be exploited to seek related molecular targets. In this review, we focus on miRNAs found to be involved in Src signalling in various cancers. We summarize recent findings on Src-related miRNAs, their target genes, mechanisms behind their interplay and their implications for cancer therapeutics.

Description: Tail-anchored (TA) proteins, a class of membrane proteins having an N-terminal cytoplasmic region anchored to the membrane by a single C-terminal transmembrane domain, are posttranslationally inserted into the endoplasmic reticulum (ER) membrane. In yeasts, the posttranslational membrane insertion is mediated by the Guided Entry of TA Proteins (GET) complex. Get3, a cytosolic ATPase, targets newly synthesized TA proteins to the ER membrane, where Get2 and Get3 constitute the Get3 receptor driving the membrane insertion. While mammalian cells employ TRC40 and WRB, mammalian homologs of Get3 and Get1, respectively, they lack the gene homologous to Get2. We recently identified calcium-modulating cyclophilin ligand (CAML) as a TRC40 receptor, indicating that CAML was equivalent to Get2 in the context of the membrane insertion. On the other hand, CAML has been well characterized as a signaling molecule that regulates various biological processes, raising the question of how the two distinct actions of CAML, the membrane insertion and the signal transduction, are assembled. In this review, we summarize recent progress of the molecular mechanism of the membrane insertion of TA proteins and discuss the possibility that CAML could sense the various signals at the ER membrane, thereby controlling TA protein biogenesis.

Description: In bacterial organisms, the oriC -independent primosome plays an essential role in replication restart after dissociation of the replication DNA-protein complex following DNA damage. PriC is a key protein component in the oriC -independent replication restart primosome. Our previous study suggested that PriC was divided into an N-terminal domain and a C-terminal domain, with the latter domain being the major contributor to single-stranded DNA (ssDNA) binding capacity. In this study, we prepared several PriC mutants in which basic and aromatic amino acid residues were mutated to alanine. Five of these residues, Arg107, Lys111, Phe118, Arg121 and Lys165 in the C-terminal domain, were shown to be involved in ssDNA binding. Moreover, we evaluated the binding of the PriC mutants to the ssDNA-binding protein (SSB) complex. Five residues, Phe118, Arg121, Arg129, Tyr152 and Arg155 in the C-terminal domain of PriC, were shown to be involved in SSB binding in the presence of ssDNA. On the basis of these results, we propose a structural model of the C-terminal domain of PriC and discuss how the interactions of PriC with SSB and ssDNA may contribute to the regulation of PriC-dependent replication restart.

Description: L -Lysine α-oxidase (LysOX) from Trichoderma viride is a homodimeric 112 kDa flavoenzyme that catalyzes the oxidative deamination of L -lysine to form α-keto--aminocaproate. LysOX severely inhibited growth of cancer cells but showed relatively low cytotoxicity for normal cells. We have determined the cDNA nucleotide sequence encoding LysOX from T. viride. The full-length cDNA consists of 2,119 bp and encodes a possible signal peptide (Met1-Arg77) and the mature protein (Ala78-Ile617). The LysOX gene have been cloned and heterologously expressed in Streptomyces lividans TK24 with the enzyme activity up to 9.8 U/ml. The enzymatic properties of the purified recombinant LysOX, such as substrate specificity and thermal stability, are same as those of native LysOX. The crystal structure of LysOX at 1.9 Å resolution revealed that the overall structure is similar to that of snake venom L -amino acid oxidase (LAAO), and the residues involved in the interaction with the amino or carboxy group of the substrate are structurally conserved. However, the entrance and the inner surface structures of the funnel to the active site, as well as the residues involved in the substrate side-chain recognition, are distinct from LAAOs. These structural differences well explain the unique substrate specificity of LysOX.

Description: For a multistep pre-targeting method using antibodies, a streptavidin mutant with low immunogenicity, termed low immunogenic streptavidin mutant No. 314 (LISA-314), was produced previously as a drug delivery tool. However, endogenous biotins (BTNs) with high affinity ( K d 〈 10 –10 M) for the binding pocket of LISA-314 prevents access of exogenous BTN-labelled anticancer drugs. In this study, we improve the binding pocket of LISA-314 to abolish its affinity for endogenous BTN species, therefore ensuring that the newly designed LISA-314 binds only artificial BTN analogue. The replacement of three amino acid residues was performed in two steps to develop a mutant termed V212, which selectively binds to 6-(5-((3a S ,4 S ,6a R )-2-iminohexahydro-1 H -thieno[3,4- d ]imidazol-4-yl)pentanamido)hexanoic acid (iminobiotin long tail, IMNtail). Surface plasmon resonance results showed that V212 has a K d value of 5.9 x 10 –7 M towards IMNtail, but no binding affinity for endogenous BTN species. This V212/IMNtail system will be useful as a novel delivery tool for anticancer therapy.

Description: A number of gene mutations are detected in cells derived from human cancer tissues, but roles of these mutations in cancer cell development are largely unknown. We examined G364R mutation of MCM4 detected in human skin cancer cells. Formation of MCM4/6/7 complex is not affected by the mutation. Consistent with this notion, the binding to MCM6 is comparable between the mutant MCM4 and wild-type MCM4. Nuclear localization of this mutant MCM4 expressed in HeLa cells supports this conclusion. Purified MCM4/6/7 complex containing the G364R MCM4 exhibited similar levels of single-stranded DNA binding and ATPase activities to the complex containing wild-type MCM4. However, the mutant complex showed only 30–50% of DNA helicase activity of the wild-type complex. When G364R MCM4 was expressed in HeLa cells, it was fractionated into nuclease-sensitive chromatin fraction, similar to wild-type MCM4. These results suggest that this mutation does not affect assembly of MCM2-7 complex on replication origins but it interferes some step at function of MCM2-7 helicase. Thus, this mutation may contribute to cancer cell development by disturbing DNA replication.

Description: The provisions in the Habitats Directive relating to protection of sites establish a triumvirate of decision-makers: administrative authority, scientific advisor and judiciary. This article examines the relationship between these decision-makers as developed in recent case law, both at a European Union (EU) and national level. It argues that reference to the goal of environmental protection obscures the allocation of power among these actors, and that to truly understand the resulting system, we must acknowledge the differing norms which motivate each of these actors. In particular, it argues that we must consider the judiciary as an actor within the decision-making process, and should examine the role of the principles of judicial review and EU law in shaping this. It highlights that there are currently conflicts within the process, and that the principles of judicial review cannot provide a successful mechanism to manage these conflicts without an explicit consideration of the values ‘hidden’ therein.

Description: One of the key issues in the current controversy over the hunting of wolves in Sweden is whether the wolf population has reached favourable conservation status (FCS). FCS is a legal concept, created and defined in law, but like many legal concepts within environmental law, can only be understood by reference to ecological concepts such as species viability. These ecological determinations in turn often require some sort of legal or policy judgment, such as how great an extinction risk is acceptable for a viable population. This article interrogates contested legal and ecological aspects of FCS and argues for how they might be applied to the Swedish wolf in potential litigation.

Description: Within the regulatory space that exists at the intersection of UK company law and environmental regulation, the business community has generated its own environmental governance initiatives to address growing anxiety about companies’ externalised risk. Yet, there is currently nothing in law to prevent companies from frequently acting inconsistently with these voluntary unilateral assurances, which has led to widespread concern that environmental values are treated as merely instrumental to the dominant idea of achieving economic benefits for the company. This article examines a specific case for the legal facilitation of binding obligations owed to the environment, which require a company to make good on its previous commitments about environmental responsibility. It seeks to demonstrate that this is possible through the common law doctrine of estoppel, which can be opened up to prevent a company from acting inconsistently with its previous statements or actions about the governance of environmental risk.

Description: The aim of the article is to shed light on the particular issue of absence of judicial dialogue between the European Court of Human Rights (ECtHR) and the Court of Justice of the EU (CJEU) in the matter of environmental rights which represents a glaring exception to the generally cooperative disposition exhibited by the two courts in other domains linked to human rights protection. The article draws on this particular absence of judicial dialogue by examining the respective patterns of judicial reasoning employed by the CJEU and the ECtHR in cases before them that involve, or have a bearing on, environmental rights (substantive and procedural). Thus, the singular tendencies discernible in the ECtHR’s progressive jurisprudence in the field of environmental rights will be compared to CJEU’s jurisprudence relevant to environmental rights with the intention of detecting certain aspects in the CJEU’s approach which could further stand to be improved following the example of ECtHR’s activist environmental jurisprudence as a viable avenue for initiating the currently missing dialogue between the two courts in the matter of environmental rights.