CydDC-mediated reductant export in Escherichia coli controls the transcriptional wiring of energy metabolism and combats nitrosative stress

The glutathione/cysteine exporter CydDC maintains redox balance in Escherichia coli. A cydD mutant strain was used to probe the influence of CydDC upon reduced thiol export, gene expression, metabolic perturbations, intracellular pH homeostasis, and tolerance to nitric oxide (NO). Loss of CydDC was found to decrease extracytoplasmic thiol levels, whereas overexpression diminished the cytoplasmic thiol content. Transcriptomic analysis revealed a dramatic up-regulation of protein chaperones, protein degradation (via phenylpropionate/phenylacetate catabolism), ?-oxidation of fatty acids, and genes involved in nitrate/nitrite reduction. 1H NMR metabolomics revealed elevated methionine and betaine and diminished acetate and NAD+ in cydD cells, which was consistent with the transcriptomics-based metabolic model. The growth rate and ?pH, however, were unaffected, although the cydD strain did exhibit sensitivity to the NO-releasing compound NOC-12. These observations are consistent with the hypothesis that the loss of CydDC-mediated reductant export promotes protein misfolding, adaptations to energy metabolism, and sensitivity to NO. The addition of both glutathione and cysteine to the medium was found to complement the loss of bd -type cytochrome synthesis in a cydD strain (a key component of the pleiotropic cydDC phenotype), providing the first direct evidence that CydDC substrates are able to restore the correct assembly of this respiratory oxidase. These data provide an insight into the metabolic flexibility of E. coli , highlight the importance of bacterial redox homeostasis during nitrosative stress, and report for the first time the ability of periplasmic low molecular weight thiols to restore haem incorporation into a cytochrome complex

Photoemission Measurement of Equilibrium Segregation at GeSi Surfaces

Photoemission spectroscopy is used to demonstrate that Ge segregates to the first atomic layer of Ge0.5Si0.5(100)2×1 and that the second layer is predominantly Si. Comparison of the resolved signals from the dimer atoms of the reconstructed (100)2×1 surfaces of Ge, Si, and equiatomic Ge‐Si alloy shows that the surface layer of the alloy is extremely Ge rich and the second layer is occupied mainly by Si atoms. This result is in good agreement with theoretical predictions

The Use of Senior Volunteers in the Care of Discharged Geriatric Patients

This article reports on a project that utilized senior volunteers in the role of health advocates for geriatric patients discharged from a hospital. The project was evaluated to determine if healthy and active seniors could make a contribution to the health and social welfare of such discharged elderly persons. The study was conducted in Montreal, Canada and funded by a federal grant from Health Canada. The research collaborators came from a 414-bed secondary care university-affiliated community hospital, a community social service agency with a mandate to respond to the needs of its frail elderly constituents, and a university-based research centr

Evolutionary Pressure on Mitochondrial Cytochrome b Is Consistent with a Role of CytbI7T Affecting Longevity during Caloric Restriction

Background: Metabolism of energy nutrients by the mitochondrial electron transport chain (ETC) is implicated in the aging process. Polymorphisms in core ETC proteins may have an effect on longevity. Here we investigate the cytochrome b (cytb) polymorphism at amino acid 7 (cytbI7T) that distinguishes human mitochondrial haplogroup H from haplogroup U. Principal Findings: We compared longevity of individuals in these two haplogroups during historical extremes of caloric intake. Haplogroup H exhibits significantly increased longevity during historical caloric restriction compared to haplogroup U(p = 0.02) while during caloric abundance they are not different. The historical effects of natural selection on the cytb protein were estimated with the software TreeSAAP using a phylogenetic reconstruction for 107 mammal taxa from all major mammalian lineages using 13 complete protein-coding mitochondrial gene sequences. With this framework, we compared the biochemical shifts produced by cytbI7T with historical evolutionary pressure on and near this polymorphic site throughout mammalian evolution to characterize the role cytbI7T had on the ETC during times of restricted caloric intake. Significance: Our results suggest the relationship between caloric restriction and increased longevity in human mitochondrial haplogroup H is determined by cytbI7T which likely enhances the ability of water to replenish the Q i binding site and decreases the time ubisemiquinone is at the Qo site, resulting in a decrease in the average production rate of radica

Crystal Field Splitting and Charge Flow in the Buckled-Dimer Reconstruction of Si(100)—2× 1

The effect of the 2×1 reconstruction on the core-electron binding energies of the outermost Si(100) layers has been determined using high-resolution photoemission data. A previously unobserved 190-meV crystal-field splitting is resolved for the up-atoms of the asymmetric surface dimers, whose average core-level shift is -400 meV. The signal from the down-atoms is clearly identified and has a shift of +220 meV. These new findings indicate a charge flow of ∼0.05e from the subsurface to the surface layers, with a substantially larger difference of ∼0.34e between the up-atoms and down-atoms in the dimer

Solution structure of a bacterial microcompartment targeting peptide and its application in the construction of an ethanol bioreactor

Targeting of proteins to bacterial microcompartments (BMCs) is mediated by an 18-amino-acid peptide sequence. Herein, we report the solution structure of the N-terminal targeting peptide (P18) of PduP, the aldehyde dehydrogenase associated with the 1,2-propanediol utilization metabolosome from Citrobacter freundii. The solution structure reveals the peptide to have a well-defined helical conformation along its whole length. Saturation transfer difference and transferred NOE NMR has highlighted the observed interaction surface on the peptide with its main interacting shell protein, PduK. By tagging both a pyruvate decarboxylase and an alcohol dehydrogenase with targeting peptides, it has been possible to direct these enzymes to empty BMCs in vivo and to generate an ethanol bioreactor. Not only are the purified, redesigned BMCs able to transform pyruvate into ethanol efficiently, but the strains containing the modified BMCs produce elevated levels of alcohol

The variable relationship between arm and hand use: a rationale for using finger magnetometry to complement wrist accelerometry when measuring daily use of the upper extremity.

Wrist-worn accelerometers are becoming more prevalent as a means to assess use of the impaired upper extremity in daily life after stroke. However, wrist accelerometry does not measure joint movements of the hand, which are integral to functional use of the upper extremity. In this study, we used a custom-built, non-obtrusive device called the manumeter to measure both arm use (via wrist accelerometry) and hand use (via finger magnetometry) of a group of unimpaired subjects while they performed twelve motor tasks at three intensities. We also gave the devices to four stroke subjects and asked them to wear them for six hours a day for one month. From the in-lab testing we found that arm use was a strong predictor of hand use for individual tasks, but that the slope of the relationship varied by up to a factor of ~12 depending on the task being performed. Consistent with this, in the daily use data collected from stroke subjects we found a broad spread in the relationship between arm and hand use. These results suggest that analyzing the spread of the relationship between daily hand and arm use will give more insight into upper extremity recovery than wrist accelerometry or finger magnetometry alone, because the spread reflects the nature of the daily tasks performed as well as the amount of upper extremity use

The Transit Light Curve Project. VIII. Six Occultations of the Exoplanet TrES-3

We present photometry of the exoplanet host star TrES-3 spanning six occultations (secondary eclipses) of its giant planet. No flux decrements were detected, leading to 99%-confidence upper limits on the planet-to-star flux ratio of 0.00024, 0.0005, and 0.00086 in the i, z, and R bands respectively. The corresponding upper limits on the planet's geometric albedo are 0.30, 0.62, and 1.07. The upper limit in the i band rules out the presence of highly reflective clouds, and is only a factor of 2-3 above the predicted level of thermal radiation from the planet.Comment: To appear in AJ [14 pages

Tensile properties of the transverse carpal ligament and carpal tunnel complex

A new sophisticated method that uses video analysis techniques together with a Maillon Rapide Delta to determine the tensile properties of the transverse carpal ligament–carpal tunnel complex has been developed. Six embalmed cadaveric specimens amputated at the mid-forearm and aged (mean (SD)): 82 (6.29) years were tested. The six hands were from three males (four hands) and one female (two hands). Using trigonometry and geometry the elongation and strain of the transverse carpal ligament and carpal arch were calculated. The cross-sectional area of the transverse carpal ligament was determined. Tensile properties of the transverse carpal ligament–carpal tunnel complex and Load–Displacement data were also obtained. Descriptive statistics, one-way ANOVA together with a post-hoc analysis (Tukey) and t-tests were incorporated. A transverse carpal ligament–carpal tunnel complex novel testing method has been developed. The results suggest that there were no significant differences between the original transverse carpal ligament width and transverse carpal ligament at peak elongation (P= 0.108). There were significant differences between the original carpal arch width and carpal arch width at peak elongation (P=0.002). The transverse carpal ligament failed either at the mid-substance or at their bony attachments. At maximum deformation the peak load and maximum transverse carpal ligament displacements ranged from 285.74 N to 1369.66 N and 7.09 mm to 18.55 mm respectively. The transverse carpal ligament cross-sectional area mean (SD) was 27.21 (3.41)mm2. Using this method the results provide useful biomechanical information and data about the tensile properties of the transverse carpal ligament–carpal tunnel complex

Electromagnetic navigation in total knee arthroplasty—a single center, randomized, single-blind study comparing the results with conventional techniques

We report on the results of a randomized study (n = 200) to compare total knee arthroplasty performed using conventional instrumentation or electromagnetic computer assisted surgical technique. 92% of navigated and 85% of conventional knees were implanted within ± 3° from neutral mechanical alignment; there was no statistically significant difference between these proportions. There was also no difference in femoral or tibial rotation assessed by CT scan. At 1 year follow up there was no statistical difference between the two groups in American Knee Society Score, Oxford Knee Scores, patient satisfaction, quality of life, hospital length of stay, complication rates or other adverse events. Tourniquet time in the navigated group was longer. Proving value for navigation in total knee arthroplasty surgery remains a challenge