Time-Resolved Cryofixation Methods for the Study of Dynamic Cellular Events by Electron Microscopy: A Review

The preservation of cells for electron microscopy by chemical fixation is a lengthy process, requiring up to 30 minutes for cytoplasmic stabilisation. This time lag enables many changes to occur in specimens so that they may not reflect their living state when they are observed in electron microscopes. Many artefacts can be avoided by using cryofixation, which freezes specimens over a period that is measured in milliseconds, so that specimens are preserved by cryoimmobilisation. This time resolution can be used to study rapid processes in biology and chemistry because, although electron microscopes cannot observe dynamic cellular events directly, processes can be arrested after known time intervals so that transient stages are preserved and a series of time-lapse steps is acquired. Some experiments have involved freezing specimens which were maintained in controlled states and others have shown results after stimulation where structural differences are seen between one millisecond and the next. The experimental techniques that have been applied prior to freezing are electrical and chemical stimulation, electrophoresis, chemical relaxation after a temperature jump, electroporation, which is analogous to relaxation after applying a radio frequency electrical field, and flash photolysis methods. This review describes the origins and application of time-resolved freezing, which integrates electron microscopy with dynamic biochemical, physiological, and ultrastructural events

Impacts of Mid-Level Biofuel Content In Gasoline on SIDI Engine-Out and Tailpipe Particulate Matter Emissions

In this work, the influences of ethanol and iso-butanol blended with gasoline on engine-out and post three-way catalyst (TWC) particle size distribution and number concentration were studied using a General Motors (GM) 2.0L turbocharged spark ignition direct injection (SIDI) engine. The engine was operated using the production engine control unit (ECU) with a dynamometer controlling the engine speed and the accelerator pedal position controlling the engine load. A TSI Fast Mobility Particle Sizer (FMPS) spectrometer was used to measure the particle size distribution in the range from 5.6 to 560 nm with a sampling rate of 1 Hz. U.S. federal certification gasoline (E0), two ethanol-blended fuels (E10 and E20), and 11.7% iso-butanol blended fuel (BU12) were tested. Measurements were conducted at 10 selected steady-state engine operation conditions. Bi-modal particle size distributions were observed for all operating conditions with peak values at particle sizes of 10 nm and 70 nm. Idle and low-speed / low-load conditions emitted higher total particle numbers than other operating conditions. At idle, the engine-out particulate matter (PM) emissions were dominated by nucleation mode particles, and the production TWC reduced these nucleation mode particles by more than 50%, while leaving the accumulation mode particle distribution unchanged. At an engine load higher than 6 bar net mean effective pressure (NMEP), accumulation mode particles dominated the engine-out particle emissions, and the TWC had little effect. Compared to the baseline gasoline (E0), E10 does not significantly change PM emissions, while E20 and BU12 both reduce PM emissions under the conditions studied. Iso-butanol was observed to impact PM emissions more than ethanol, with up to 50% reductions at some conditions. In this paper, issues related to PM measurement using the FMPS are also discussed. While some uncertainties are due to engine variation, the FMPS must be carefully maintained in order to achieve repeatable measurement results

Criteria for and Appropriateness of Renal Transplantation in Elderly Patients With End-Stage Renal Disease : A Literature Review and Position Statement on Behalf of the European Renal Association-European Dialysis and Transplant Association Descartes Working Group and European Renal Best Practice

During the last 20 years, waiting lists for renal transplantation (RT) have grown significantly older. However, elderly patients (ie ≥65 years of age) are still more rarely referred or accepted to waiting lists and, if enlisted, have less chances of actually receiving a kidney allograft, than younger counterparts. In this review, we looked at evidence for the benefits and risks of RT in the elderly trying to answer the following questions: Should RT be advocated for elderly patients? What should be the criteria to accept elderly patients on the waiting list for RT? What strategies might be used to increase the rate of RT in waitlisted elderly candidates? For selected elderly patients, RT was shown to be superior to dialysis in terms of patient survival. Virtually all guidelines recommend that patients should not be deemed ineligible for RT based on age alone, although a short life expectancy generally might preclude RT. Concerning the assessment of comorbidities in the elderly, special attention should be paid to cardiac evaluation and screening for malignancy. Comorbidity scores and frailty assessment scales might help the decision making on eligibility. Psychosocial issues should also be evaluated. To overcome the scarcity of organ donors, elderly RT candidates should be encouraged to consider expanded criteria donors and living donors, as alternatives to deceased standard criteria donors. It has been demonstrated that expanded criteria donor RT in patients 60 years or older is associated with higher survival rates than remaining on dialysis, whereas living donor RT is superior to all other options.Peer reviewe

Effects of Intermediate Ethanol Blends on Legacy Vehicles and Small Non‑Road Engines, Report 1 - Updated

In summer 2007, the U.S. Department of Energy (DOE) initiated a test program to evaluate the potential impacts of intermediate ethanol blends on legacy vehicles and other engines. The purpose of the test program is to assess the viability of using intermediate blends as a contributor to meeting national goals in the use of renewable fuels. Through a wide range of experimental activities, DOE is evaluating the effects of E15 and E20--gasoline blended with 15 and 20% ethanol--on tailpipe and evaporative emissions, catalyst and engine durability, vehicle driveability, engine operability, and vehicle and engine materials. This first report provides the results available to date from the first stages of a much larger overall test program. Results from additional projects that are currently underway or in the planning stages are not included in this first report. The purpose of this initial study was to quickly investigate the effects of adding up to 20% ethanol to gasoline on the following: (1) Regulated tailpipe emissions for 13 popular late model vehicles on a drive cycle similar to real-world driving and 28 small non-road engines (SNREs) under certification or typical in use procedures. (2) Exhaust and catalyst temperatures of the same vehicles under more severe conditions. (3) Temperature of key engine components of the same SNREs under certification or typical in-use conditions. (4) Observable operational issues with either the vehicles or SNREs during the course of testing. As discussed in the concluding section of this report, a wide range of additional studies are underway or planned to consider the effects of intermediate ethanol blends on materials, emissions, durability, and driveability of vehicles, as well as impacts on a wider range of nonautomotive engines, including marine applications, snowmobiles, and motorcycles. Section 1 (Introduction) gives background on the test program and describes collaborations with industry and agencies to date. Section 2 (Experimental Setup) provides details concerning test fuels, vehicle and SNRE selection, and test methods used to conduct the studies presented in this report. Section 3 (Results and Discussion) summarizes the vehicle and SNRE studies and presents data from testing completed to date. Section 4 (Next Steps) describes planned future activities. The appendixes provide test procedure details, vehicle and SNRE emissions standards, analysis details, and additional data and tables from vehicle and SNRE tests

Judicious Ligand Design in Ruthenium Polypyridyl CO2 Reduction Catalysts to Enhance Reactivity by Steric and Electronic Effects

A series of RuII polypyridyl complexes of the structural design [RuII(R−tpy)(NN)(CH3CN)]2+ (R−tpy=2,2′:6′,2′′-terpyridine (R=H) or 4,4′,4′′-tri-tert-butyl-2,2′:6′,2′′-terpyridine (R=tBu); NN=2,2′-bipyridine with methyl substituents in various positions) have been synthesized and analyzed for their ability to function as electrocatalysts for the reduction of CO2 to CO. Detailed electrochemical analyses establish how substitutions at different ring positions of the bipyridine and terpyridine ligands can have profound electronic and, even more importantly, steric effects that determine the complexes’ reactivities. Whereas electron-donating groups para to the heteroatoms exhibit the expected electronic effect, with an increase in turnover frequencies at increased overpotential, the introduction of a methyl group at the ortho position of NN imposes drastic steric effects. Two complexes, [RuII(tpy)(6-mbpy)(CH3CN)]2+ (trans-[3]2+; 6-mbpy=6-methyl-2,2′-bipyridine) and [RuII(tBu−tpy)(6-mbpy)(CH3CN)]2+ (trans-[4]2+), in which the methyl group of the 6-mbpy ligand is trans to the CH3CN ligand, show electrocatalytic CO2 reduction at a previously unreactive oxidation state of the complex. This low overpotential pathway follows an ECE mechanism (electron transfer–chemical reaction–electron transfer), and is a direct result of steric interactions that facilitate CH3CN ligand dissociation, CO2 coordination, and ultimately catalytic turnover at the first reduction potential of the complexes. All experimental observations are rigorously corroborated by DFT calculations

Accelerating Policy Decisions to Adopt Haemophilus influenzae Type b Vaccine: A Global, Multivariable Analysis

Jessica Shearer and colleagues analyze data from 147 countries to identify factors that influence the time taken to introduce routine vaccination against Haemophilus influenzae type b (Hib)

Should the Arteriovenous Fistula Be Created before Starting Dialysis?: A Decision Analytic Approach

Background: An arteriovenous fistula (AVF) is considered the vascular access of choice, but uncertainty exists about the\ud optimal time for its creation in pre-dialysis patients. The aim of this study was to determine the optimal vascular access\ud referral strategy for stage 4 (glomerular filtration rate ,30 ml/min/1.73 m2) chronic kidney disease patients using a decision\ud analytic framework.\ud Methods: A Markov model was created to compare two strategies: refer all stage 4 chronic kidney disease patients for an\ud AVF versus wait until the patient starts dialysis. Data from published observational studies were used to estimate the\ud probabilities used in the model. A Markov cohort analysis was used to determine the optimal strategy with life expectancy\ud and quality adjusted life expectancy as the outcomes. Sensitivity analyses, including a probabilistic sensitivity analysis, were\ud performed using Monte Carlo simulation.\ud Results: The wait strategy results in a higher life expectancy (66.6 versus 65.9 months) and quality adjusted life expectancy\ud (38.9 versus 38.5 quality adjusted life months) than immediate AVF creation. It was robust across all the parameters except\ud at higher rates of progression and lower rates of ischemic steal syndrome.\ud Conclusions: Early creation of an AVF, as recommended by most guidelines, may not be the preferred strategy in all predialysis\ud patients. Further research on cost implications and patient preferences for treatment options needs to be done\ud before recommending early AVF creation

Higher Level Phylogeny and the First Divergence Time Estimation of Heteroptera (Insecta: Hemiptera) Based on Multiple Genes

Heteroptera, or true bugs, are the largest, morphologically diverse and economically important group of insects with incomplete metamorphosis. However, the phylogenetic relationships within Heteroptera are still in dispute and most of the previous studies were based on morphological characters or with single gene (partial or whole 18S rDNA). Besides, so far, divergence time estimates for Heteroptera totally rely on the fossil record, while no studies have been performed on molecular divergence rates. Here, for the first time, we used maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) with multiple genes (18S rDNA, 28S rDNA, 16S rDNA and COI) to estimate phylogenetic relationships among the infraorders, and meanwhile, the Penalized Likelihood (r8s) and Bayesian (BEAST) molecular dating methods were employed to estimate divergence time of higher taxa of this suborder. Major results of the present study included: Nepomorpha was placed as the most basal clade in all six trees (MP trees, ML trees and Bayesian trees of nuclear gene data and four-gene combined data, respectively) with full support values. The sister-group relationship of Cimicomorpha and Pentatomomorpha was also strongly supported. Nepomorpha originated in early Triassic and the other six infraorders originated in a very short period of time in middle Triassic. Cimicomorpha and Pentatomomorpha underwent a radiation at family level in Cretaceous, paralleling the proliferation of the flowering plants. Our results indicated that the higher-group radiations within hemimetabolous Heteroptera were simultaneously with those of holometabolous Coleoptera and Diptera which took place in the Triassic. While the aquatic habitat was colonized by Nepomorpha already in the Triassic, the Gerromorpha independently adapted to the semi-aquatic habitat in the Early Jurassic