Computational design of Periplasmic binding protein biosensors guided by molecular dynamics.

Periplasmic binding proteins (PBPs) are bacterial proteins commonly used as scaffolds for substrate-detecting biosensors. In these biosensors, effector proteins (for example fluorescent proteins) are inserted into a PBP such that the effector protein's output changes upon PBP-substate binding. The insertion site is often determined by comparison of PBP apo/holo crystal structures, but random insertion libraries have shown that this can miss the best sites.

High laser fluence ITO coatings utilizing a Fabry-Perot thin film filter to reduce effective absorption.

Laser-induced damage threshold of transparent conductors, such as Indium Tin Oxide (ITO), is limited by their high optical absorption due to free carriers. However, the effective absorption of a transparent conductor thin film can be reduced by an order of magnitude, without changing the electrical characteristics of the film, when placed in a low electric field section of a multilayer coating. A Fabry-Perot thin film interference filter has both high transmittance and low electric field positions, so it is an ideal thin film structure for this application.

Affordable and robust phenotyping framework to analyse root system architecture of soil-grown plants.

The phenotypic analysis of root system growth is important to inform efforts to enhance plant resource acquisition from soils; however, root phenotyping remains challenging because of the opacity of soil, requiring systems that facilitate root system visibility and image acquisition. Previously reported systems require costly or bespoke materials not available in most countries, where breeders need tools to select varieties best adapted to local soils and field conditions. Here, we report an affordable soil-based growth (rhizobox) and imaging system to phenotype root development in glasshouses or shelters.

The impact of the rhizobia-legume symbiosis on host root system architecture.

Legumes form symbioses with rhizobia to fix N2 in root nodules to supplement their nitrogen (N) requirements. Many studies have shown how symbioses affect the shoot, but far less is understood about how they modify root development and root system architecture (RSA). RSA is the distribution of roots in space and over time.

Inducible reporter/driver lines for the Arabidopsis root with intrinsic reporting of activity state.

Cell-, tissue- or organ-specific inducible expression systems are powerful tools for functional analysis of changes to the pattern, level or timing of gene expression. However, plant researchers lack standardised reagents that promote reproducibility across the community. Here, we report the development and functional testing of a Gateway-based system for quantitatively, spatially and temporally controlling inducible gene expression in Arabidopsis that overcomes several drawbacks of the legacy systems.

Pheno-Deep Counter: a unified and versatile deep learning architecture for leaf counting.

Direct observation of morphological plant traits is tedious and a bottleneck for high-throughput phenotyping. Hence, interest in image-based analysis is increasing, with the requirement for software that can reliably extract plant traits, such as leaf count, preferably across a variety of species and growth conditions. However, current leaf counting methods do not work across species or conditions and therefore may lack broad utility.

The Phosphate Fast-Responsive Genes and Affect Phosphocholine and Phosphoethanolamine Content.

Phosphate starvation-mediated induction of the HAD-type phosphatases PPsPase1 (AT1G73010) and PECP1 (AT1G17710) has been reported in Arabidopsis (). However, little is known about their in vivo function or impact on plant responses to nutrient deficiency. The preferences of PPsPase1 and PECP1 for different substrates have been studied in vitro but require confirmation in planta.

Extreme Dynamics in the BamA β-Barrel Seam.

BamA is an essential component of the β-barrel assembly machine (BAM) that is responsible for insertion and folding of β-barrel outer membrane proteins (OMPs) in Gram-negative bacteria. BamA is an OMP itself, and its β-barrel transmembrane domain is thought to catalyze OMP insertion and folding, although the molecular mechanism remains poorly understood. Crystal structures of BamA and complementary molecular dynamics simulations have shown that its β-barrel seam (the interface between the first and last barrel strands) is destabilized.

Flexibility in the Periplasmic Domain of BamA Is Important for Function.

The β-barrel assembly machine (BAM) mediates the biogenesis of outer membrane proteins (OMPs) in Gram-negative bacteria. BamA, the central BAM subunit composed of a transmembrane β-barrel domain linked to five polypeptide transport-associated (POTRA) periplasmic domains, is thought to bind nascent OMPs and undergo conformational cycling to catalyze OMP folding and insertion. One model is that conformational flexibility between POTRA domains is part of this conformational cycling.

Longitudinal zonation pattern in Arabidopsis root tip defined by a multiple structural change algorithm.

Background and Aims The Arabidopsis thaliana root is a key experimental system in developmental biology. Despite its importance, we are still lacking an objective and broadly applicable approach for identification of number and position of developmental domains or zones along the longitudinal axis of the root apex or boundaries between them, which is essential for understanding the mechanisms underlying cell proliferation, elongation and differentiation dynamics during root development. Methods We used a statistics approach, the multiple structural change algorithm (MSC), for estimating the number and position of developmental transitions in the growing portion of the root apex.

Small Angle X-ray Scattering (SAXS) Characterization of the POTRA Domains of BamA.

BamA is the central component of the BAM complex and contains a C-terminal β-barrel domain embedded in the outer membrane, and a soluble, periplasmic domain, made out of five polypeptide transport associated (POTRA) motifs. Structural characterization of the POTRA domains was carried out by a combination of crystallographic, NMR and solution Small Angle X-ray Scattering (SAXS) approaches. Despite its limited resolution, SAXS is an excellent complement to NMR and crystallography.

Chromium chloride increases insulin-stimulated glucose uptake in the perfused rat hindlimb.

Aim: To determine the effect of chromium chloride (CrCl3 ) on healthy skeletal muscle glucose uptake in the absence and presence of submaximal insulin using the rat hindlimb perfusion technique.

Methods: Sprague-Dawley rats were randomly assigned to an experimental group: basal (Bas), chromium chloride (Cr), submaximal insulin (sIns) or chromium chloride plus submaximal insulin (Cr-sIns).

Results: Insulin significantly increased [H(3)]-2 deoxyglucose (2-DG) uptake in the gastrocnemius muscles.

An amino acid mixture is essential to optimize insulin-stimulated glucose uptake and GLUT4 translocation in perfused rodent hindlimb muscle.

The purpose of this study was to investigate whether an amino acid mixture increases glucose uptake across perfused rodent hindlimb muscle in the presence and absence of a submaximal insulin concentration, and if the increase in glucose uptake is related to an increase in GLUT4 plasma membrane density. Sprague-Dawley rats were separated into one of four treatment groups: basal, amino acid mixture, submaximal insulin, or amino acid mixture with submaximal insulin. Glucose uptake was greater for both insulin-stimulated treatments compared with the non-insulin-stimulated treatment groups but amino acids only increased glucose uptake in the presence of insulin.

Genetic and molecular basis of nonhost disease resistance: complex, yes; silver bullet, no.

Nonhost resistance (NHR), in which a successful pathogen on some plants fails to overcome host barriers on others, has attracted much attention owing to its potential for robust crop improvement. Recent advances reveal that a multitude of underlying mechanisms contribute to NHR, ranging from components shared with recognition-based defenses up to recessive susceptibility factors involved in plant primary metabolism. Most NHR appears multi-factorial and quantitative.

Structural and biochemical insights into the mechanism of fosfomycin phosphorylation by fosfomycin resistance kinase FomA.

We present here the crystal structures of fosfomycin resistance protein (FomA) complexed with MgATP, with ATP and fosfomycin, with MgADP and fosfomycin vanadate, with MgADP and the product of the enzymatic reaction, fosfomycin monophosphate, and with ADP at 1.87, 1.58, 1.

Postexercise carbohydrate-protein supplementation improves subsequent exercise performance and intracellular signaling for protein synthesis.

Postexercise carbohydrate-protein (CHO + PRO) supplementation has been proposed to improve recovery and subsequent endurance performance compared to CHO supplementation. This study compared the effects of a CHO + PRO supplement in the form of chocolate milk (CM), isocaloric CHO, and placebo (PLA) on recovery and subsequent exercise performance. Ten cyclists performed 3 trials, cycling 1.

A low carbohydrate-protein supplement improves endurance performance in female athletes.

The purpose of this study was to investigate if a low mixed carbohydrate (CHO) plus moderate protein (PRO) supplement, provided during endurance exercise, would improve time to exhaustion (TTE) in comparison to a traditional 6% CHO supplement. Fourteen (n = 14) trained female cyclists and triathletes cycled on 2 separate occasions for 3 hours at intensities varying between 45 and 70% VO2max, followed by a ride to exhaustion at an intensity approximating the individual's ventilatory threshold average 75.06% VO2max.

Pseudomonas sax genes overcome aliphatic isothiocyanate-mediated non-host resistance in Arabidopsis.

Most plant-microbe interactions do not result in disease; natural products restrict non-host pathogens. We found that sulforaphane (4-methylsulfinylbutyl isothiocyanate), a natural product derived from aliphatic glucosinolates, inhibits growth in Arabidopsis of non-host Pseudomonas bacteria in planta. Multiple sax genes (saxCAB/F/D/G) were identified in Pseudomonas species virulent on Arabidopsis.

The effect of a low carbohydrate beverage with added protein on cycling endurance performance in trained athletes.

Ingesting carbohydrate plus protein during prolonged variable intensity exercise has demonstrated improved aerobic endurance performance beyond that of a carbohydrate supplement alone. The purpose of the present study was to determine if a supplement containing a mixture of different carbohydrates (glucose, maltodextrin, and fructose) and a moderate amount of protein given during endurance exercise would increase time to exhaustion (TTE), despite containing 50% less total carbohydrate than a carbohydrate-only supplement. We also sought post priori to determine if there was a difference in effect based on percentage of ventilatory threshold (VT) at which the subjects cycled to exhaustion.

Plant development: confused root cells have mixed identity in SCHIZORIZA.

Recently discovered regulators of asymmetric cell division highlight differences in the mechanisms responsible for cell fate segregation in plants and animals.

Arabidopsis DUO POLLEN3 is a key regulator of male germline development and embryogenesis.

Male germline development in angiosperms produces the pair of sperm cells required for double fertilization. A key regulator of this process in Arabidopsis thaliana is the male germline-specific transcription factor DUO POLLEN1 (DUO1) that coordinates germ cell division and gamete specification. Here, we uncover the role of DUO3, a nuclear protein that has a distinct, but overlapping role with DUO1 in male germline development.

Gibberellin signaling in the endodermis controls Arabidopsis root meristem size.

Plant growth is driven by cell proliferation and elongation. The hormone gibberellin (GA) regulates Arabidopsis root growth by controlling cell elongation, but it is currently unknown whether GA also controls root cell proliferation. Here we show that GA biosynthetic mutants are unable to increase their cell production rate and meristem size after germination.