Deconstructing mammalian reproduction: using knockouts to define fertility pathways

Reproduction is the sine qua non for the propagation of species and continuation of life. It is a complex biological process that is regulated by multiple factors during the reproductive life of an organism. Over the past decade, the molecular mechanisms regulating reproduction in mammals have been rapidly unraveled by the study of a vast number of mouse gene knockouts with impaired fertility. The use of reverse genetics to generate null mutants in mice through targeted disruption of specific genes has enabled researchers to identify essential regulators of spermatogenesis and oogenesis in vivo and model human disorders affecting reproduction. This review focuses on the merits, utility, and the variations of the knockout technology in studies of reproduction in mammals.     

Toxic groundwater contaminants: an overlooked contributor to urban stream syndrome?

Screening for common groundwater contaminants was performed along eight urban stream reaches (100s-1000s of m) at approximately 25-75 cm below the streambeds. Four sites had known or suspected chlorinated-solvent plumes; otherwise no groundwater contamination was known previously. At each site, between 5 and 22 contaminants were detected at levels above guideline concentrations for the preservation of aquatic life, while several others were detected at lower levels, but which may still indicate some risk. Contaminants of greatest concern include numerous metals (Cd, Zn, Al, Cu, Cr, U), arsenic, various organics (chlorinated and petroleum), nitrate and ammonium, and chloride (road salt likely), with multiple types occurring at each site and often at the same sampling location. Substantial portions of the stream reaches (from 40 to 88% of locations sampled) possessed one or more contaminants above guidelines. These findings suggest that this diffuse and variable-composition urban groundwater contamination is a toxicity concern for all sites and over a large portion of each study reach. Synergistic toxicity, both for similar and disparate compounds, may also be important. We conclude that groundwater contaminants should be considered a genuine risk to urban stream aquatic ecosystems, specifically benthic organisms, and may contribute to urban stream syndrome.     

Projecting water withdrawal and supply for future decades in the U.S. under climate change scenarios

The sustainability of water resources in future decades is likely to be affected by increases in water demand due to population growth, increases in power generation, and climate change. This study presents water withdrawal projections in the United States (U.S.) in 2050 as a result of projected population increases and power generation at the county level as well as the availability of local renewable water supplies. The growth scenario assumes the per capita water use rate for municipal withdrawals to remain at 2005 levels and the water use rates for new thermoelectric plants at levels in modern closed-loop cooling systems. In projecting renewable water supply in future years, median projected monthly precipitation and temperature by sixteen climate models were used to derive available precipitation in 2050 (averaged over 2040-2059). Withdrawals and available precipitation were compared to identify regions that use a large fraction of their renewable local water supply. A water supply sustainability risk index that takes into account additional attributes such as susceptibility to drought, growth in water withdrawal, increased need for storage, and groundwater use was developed to evaluate areas at greater risk. Based on the ranking by the index, high risk areas can be assessed in more mechanistic detail in future work.     

Comparison of a real-time PCR method with a culture method for the detection of Salmonella enterica serotype enteritidis in naturally contaminated environmental samples from integrated poultry houses

Conventional culture methods have traditionally been considered the "gold standard" for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis-specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis-specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies to improve food safety.     

Bacterial attachment to RO membranes surface-modified by concentration-polarization-enhanced graft polymerization

Concentration polarization-enhanced radical graft polymerization, a facile surface modification technique, was examined as an approach to reduce bacterial deposition onto RO membranes and thus contribute to mitigation of biofouling. For this purpose an RO membrane ESPA-1 was surface-grafted with a zwitterionic and negatively and positively charged monomers. The low monomer concentrations and low degrees of grafting employed in modifications moderately reduced flux (by 20-40%) and did not affect salt rejection, yet produced substantial changes in surface chemistry, charge and hydrophilicity. The propensity to bacterial attachment of original and modified membranes was assessed using bacterial deposition tests carried out in a parallel plate flow setup using a fluorescent strain of Pseudomonas fluorescens. Compared to unmodified ESPA-1 the deposition (mass transfer) coefficient was significantly increased for modification with the positively charged monomer. On the other hand, a substantial reduction in bacterial deposition rates was observed for membranes modified with zwitterionic monomer and, still more, with very hydrophilic negatively charged monomers. This trend is well explained by the effects of surface charge (as measured by ζ-potential) and hydrophilicity (contact angle). It also well correlated with force distance measurements by AFM using surrogate spherical probes with a negative surface charge mimicking the bacterial surface. The positively charged surface showed a strong hysteresis with a large adhesion force, which was weaker for unmodified ESPA-1 and still weaker for zwitterionic surface, while negatively charged surface showed a long-range repulsion and negligible hysteresis. These results demonstrate the potential of using the proposed surface- modification approach for varying surface characteristics, charge and hydrophilicity, and thus minimizing bacterial deposition and potentially reducing propensity biofouling.     

PMF analysis of wide-range particle size spectra collected on a major highway

Particle number concentration data have been collected on a very busy road in central London (Marylebone Road). Continuous size distributions from 15 nm to 10 μm diameter, collected over 21 days, were analyzed using positive matrix factorization which identified 10 factors, five of which were observed to make major contributions (greater than 8%) to either the total number or volume of particulate matter. The sources associated with each factor were identified from the size distribution, directional association, diurnal variation and their relationship to meteorological pollution and traffic volume variables. The factors related to the emissions on Marylebone Road accounted for 40.5% of particle volume and 71.9% of particle number. These comprised nucleation mode exhaust particles (3.6% of total volume and 27.4% of total number), solid mode exhaust particles (18.8% of total volume and 38.0% of total number), brake dust (13.7% of total volume and 1.7% of total number and resuspension (4.4% of total volume and 4.8% of total number). Another six factors were associated with the urban background accounting for 59.5% of total volume and 28.2% of total particle number count. The method is extremely successful at separating the components of on-road emissions including brake wear and resuspension.     

The ZEBRA electric vehicle battery: power and energy improvements

Vehicle trials with the first sodium/nickel chloride ZEBRA batteries indicated that the pulse power capability of the battery needed to be improved towards the end of the discharge. A research programme led to several design changes to improve the cell which, in combination, have improved the power of the battery to greater than 150 W kg⁻¹ at 80% depth of discharge. Bench and vehicle tests have established the stability of the high power battery over several years of cycling. The gravimetric energy density of the first generation of cells was less than 100 Wh kg⁻¹. Optimisation of the design has led to a cell with a specific energy of 120 Wh kg⁻¹ or 86 Wh kg⁻¹ for a 30 kWh battery. Recently, the cell chemistry has been altered to improve the useful capacity. The cell is assembled in the over-discharged state and during the first charge the following reactions occur: at 1.6 V: Al+4NaCl=NaAlCl₄+3Na; at 2.35 V: Fe+2NaCl=FeCl₂+2Na; at 2.58 V: Ni+2NaCl=NiCl₂+2 Na. The first reaction serves to prime the negative sodium electrode but occurs at too low a voltage to be of use in providing useful capacity. By minimising the aluminium content more NaCl is released for the main reactions to improve the capacity of the cell. This, and further composition optimisation, have resulted in cells with specific energies in excess of 140 Wh kg⁻¹, which equates to battery energies>100 Wh kg⁻¹. The present production battery, as installed in a Mercedes Benz A class electric vehicle, gives a driving range of 205 km (128 miles) in city and hill climbing. The cells with improved capacity will extend the practical driving range to beyond 240 km (150 miles).     

Viscosity variation considering cavitation in a journal bearing lubricant containing additives

The variation in viscosity with temperature is important in lubrication: this was studied by using a viscosity—film thickness relation. This is justified since it has been verified experimentally that the highest temperature occurs in zones where the film thickness is lowest. The use of additives was considered by using a micropolar fluid model. The geometry considered was that of an infinitely long journal bearing considering cavitation. There is an increase in the load capacity when additives are used. It was established theoretically that the use of additives increases the operating temperature range in bearings.     

Fluid–Solid Reaction Kinetics for Solids of Non-basic Geometries: Application of the Law of Additive Times in Combination with the Shape-Factor Method

Metallurgical and Materials Transactions B - Fluid–solid reactions often occur under the mixed rate control of chemical and mass transfer processes. The results of this work demonstrate that...     

Diffuse reflectance spectroscopy of particulate systems: a comparative study of discrete models

A comparative study of statistical particle model theory of diffuse reflectance has been made. Theories have been applied to six particulate samples having different optical characteristics and average particle sizes that vary from 42 to 106 mum. We made an overall assessment of each theoretical model by determining the CIELAB color difference using experimentally measured and theoretically predicted diffuse reflectance spectra in the visible spectral range. We discuss the performance ratings of the models of other researchers and discovered numerous typographical errors in Fresnel reflection coefficient expressions. We provide the correct versions for these expressions.