Ultrasensitive Label-Free Detection of Protein-Membrane Interaction Exemplified by Toxin-Liposome Insertion.

Measuring the high-affinity binding of proteins to liposome membranes remains a challenge. Here, we show an ultrasensitive and direct detection of protein binding to liposome membranes using high throughput second harmonic scattering (SHS). Perfringolysin O (PFO), a pore-forming toxin, with a highly membrane selective insertion into cholesterol-rich membranes is used. PFO inserts only into liposomes with a cholesterol concentration >30%. Twenty mole-percent cholesterol results in neither SHS-signal deviation nor pore formation as seen by cryo-electron microscopy of PFO and liposomes. PFO inserts into cholesterol-rich membranes of large unilamellar vesicles in an aqueous solution with Kd = (1.5 ± 0.2) × 10-12 M. Our results demonstrate a promising approach to probe protein-membrane interactions below sub-picomolar concentrations in a label-free and noninvasive manner on 3D systems. More importantly, the volume of protein sample is ultrasmall (<10 μL). These findings enable the detection of low-abundance proteins and their interaction with membranes

Optical studies of strain and defect distribution in semipolar (1(1)over-bar01) GaN on patterned Si substrates

Formation of defects in semipolar ( 11¯01 )-oriented GaN layers grown by metal-organic chemical vapor deposition on patterned Si (001) substrates and their effects on optical properties were investigated by steady-state and time-resolved photoluminescence (PL) and spectrally and spatially resolved cathodoluminescence (CL). Near-band edge emission is found to be dominant in the c +-wings of semipolar ( 11¯01 )GaN, which are mainly free from defect-related emission lines, while the c – wings contain a large number of basal stacking faults. When the advancing c+ and c — fronts meet to coalesce into a continuous film, the existing stacking faults contained in c — wings continue to propagate in the direction perpendicular to the c-axis and, as a result, the region dominated by stacking fault emission is extended to the film surface.Additional stacking faults are observed within the c+ wings, where the growing c+ wings of GaN are in contact with the SiO2 masking layer. Out-diffusion of oxygen/silicon species and concentration of strain near the contact region are considered as possible causes of the stacking fault formation. CL linescans performed along the surface and across the thickness of the non-coalesced and coalesced layers revealed that, while most of the material in the near-surface region of the non-coalesced layers is relaxed, coalescence results in nonuniform strain distribution over the layer surface. Red-shifted near-band-edge emission from the near-surface region indicates tensile stress near the surface of a coalesced layer, reaching a value of 0.3 GPa. The regions near the GaN/AlN/Si(111) interface show slightly blue shifted, broadened near-band-edge emission, which is indicative of a high concentration of free carriers possibly due to incorporation of shallow-donor impurities (Si and/or O) from the substrate or SiO2 mask. Steady-state and time-resolved PL results indicate that semipolar ( 11¯01 )GaN on patterned Si exhibits optical properties (PL intensity and carrier lifetimes) approaching to those of the state-of-the-art c-plane GaN grown using in situ SiNx nanonetwork mask on c-plane sapphire. Long PL lifetimes (∼2 ns) for the ( 11¯01 )GaN layers show that the semipolar material holds promise for light emitting and detecting devices

Isolated recessive nail dysplasia caused by <i>FZD6 </i>mutations:report of three families and review of the literature

Congenital abnormalities of the nail are rare conditions that are most frequently associated with congenital ectodermal syndromes involving several of the epidermal appendages including the skin, teeth, hair and nails. Isolated recessive nail dysplasia (IRND) is much rarer but has recently been recognized as a condition resulting in 20‐nail dystrophy in the absence of other cutaneous or extracutaneous findings. A few case reports have identified mutations in the Frizzled 6 (FZD6) gene in families presenting with abnormal nails consistent with IRND. These reports have highlighted the role of Wnt–FZD signalling in the process of nail formation. We report three families presenting with features of IRND, in whom we identified mutations in FZD6, including one previously unreported mutation

Impact of active layer design on InGaN radiative recombination coefficient and LED performance

The relative roles of radiative and nonradiative processes and the polarization field on the light emission from blue (∼425 nm) InGaN light emitting diodes (LEDs) have been studied. Single and multiple double heterostructure (DH) designs have been investigated with multiple DH structures showing improved efficiencies. Experimental results supported by numerical simulations of injection dependent electron and hole wavefunction overlap and the corresponding radiative recombination coefficients suggest that increasing the effective active region thickness by employing multiple InGaN DH structures separated by thin and low barriers is promising for LEDs with high efficiency retention at high injection. The use of thin and low barriers is crucial to enhance carrier transport across the active region. Although increasing the single DH thickness from 3 to 6 nm improves the peak external quantum efficiency (EQE) by nearly 3.6 times due to increased density of states and increased emitting volume, the internal quantum efficiency (IQE) suffers a loss of nearly 30%. A further increase in the DH thickness to 9 and 11 nm results in a significantly slower rate of increase of EQE with current injection and lower peak EQE values presumably due to degradation of the InGaN material quality and reduced electron-hole spatial overlap. Increasing the number of 3 nm DH active regions separated by thin (3 nm) In0.06Ga0.94N barriers improves EQE, while maintaining high IQE (above 95% at a carrier concentration of 1018 cm−3) and showing negligible EQE degradation up to 550 A/cm2 in 400 × 400 μm2 devices due to increased emitting volume and high radiative recombination coefficients and high IQE. Time-resolved photoluminescence measurements revealed higher radiative recombination rates with increasing excitation due to screening of the internal field and enhanced electron and hole overlap at higher injection levels. To shed light on the experimental observations, the effect of free-carrier screening on the polarization field at different injection levels and the resulting impact on the quantum efficiency were investigated by numerical simulations

Excitonic recombination dynamics in non-polar GaN/AlGaN quantum wells

The optical properties of GaN/Al0.15Ga0.85N multiple quantum wells are examined in 8 K–300 K temperature range. Both polarized CW and time resolved temperature-dependent photoluminescence experiment are performed so that we can deduce the relative contributions of the non-radiative and radiative recombination processes. From the calculation of the proportion of the excitonic population having wave vector in the light cone, we can deduce the variation of the radiative decay time with temperature. We find part of the excitonic population to be localized in concert with the report of Corfdir et al. (Jpn. J. Appl. Phys., Part 2 52, 08JC01 (2013)) in case of a-plane quantum wells

Ethanol production from xylose by pichia stipitis NRRL Y-7124 in a stirred tank bioreactor

The ethanol production by Pichia stipitis was evaluated in a stirred tank bioreactor using semidefined medium containing xylose (90.0 g/l) as the main carbon source. Experimental assays were performed according to a 22 full factorial design to evaluate the influence of aeration (0.25 to 0.75 vvm) and agitation (150 to 250 rpm) conditions on ethanol production. In the studied range of values, the agitation increase and aeration decrease favored ethanol production, which was maximum (26.7 g/l) using 250 rpm and 0.25 vvm, conditions that gave a volumetric oxygen transfer coefficient (kLa value) of 4.9 h-1. Under these conditions, the ethanol yield factor, ethanol productivity, and the process efficiency were 0.32 g/g, 0.32 g/l.h, and 63%, respectively. These results are promising and contribute to the development of a suitable process for ethanol production from xylose by Pichia stipitis.The authors gratefully acknowledge Santander, Fapesp, Capes, and CNPq (Brazil)

Biallelic Mutations in ADPRHL2, Encoding ADP-Ribosylhydrolase 3, Lead to a Degenerative Pediatric Stress-Induced Epileptic Ataxia Syndrome.

ADP-ribosylation, the addition of poly-ADP ribose (PAR) onto proteins, is a response signal to cellular challenges, such as excitotoxicity or oxidative stress. This process is catalyzed by a group of enzymes referred to as poly(ADP-ribose) polymerases (PARPs). Because the accumulation of proteins with this modification results in cell death, its negative regulation restores cellular homeostasis: a process mediated by poly-ADP ribose glycohydrolases (PARGs) and ADP-ribosylhydrolase proteins (ARHs). Using linkage analysis and exome or genome sequencing, we identified recessive inactivating mutations in ADPRHL2 in six families. Affected individuals exhibited a pediatric-onset neurodegenerative disorder with progressive brain atrophy, developmental regression, and seizures in association with periods of stress, such as infections. Loss of the Drosophila paralog Parg showed lethality in response to oxidative challenge that was rescued by human ADPRHL2, suggesting functional conservation. Pharmacological inhibition of PARP also rescued the phenotype, suggesting the possibility of postnatal treatment for this genetic condition

Random UV-C mutagenesis of Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 to improve anaerobic growth on lignocellulosic sugars

Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 was mutagenized using UV-C irradiation to produce yeast strains for anaerobic conversion of lignocellulosic sugars to ethanol. UV-C irradiation potentially produces large numbers of random mutations broadly and uniformly over the whole genome to generate unique strains. Wild-type cultures of S. stipitis NRRL Y-7124 were subjected to UV-C (234 nm) irradiation targeted at approximately 40% cell survival. When surviving cells were selected in sufficient numbers via automated plating strategies and cultured anaerobically on xylose medium for 5 months at 28°C, five novel mutagenized S. stipitis strains were obtained. Variable number tandem repeat analysis revealed that mutations had occurred in the genome, which may have produced genes that allowed the anaerobic utilization of xylose. The mutagenized strains were capable of growing anaerobically on xylose/glucose substrate with higher ethanol production during 250- to 500-h growth than a Saccharomyces cerevisiae yeast strain that is the standard for industrial fuel ethanol production. The S. stipitis strains resulting from this intense multigene mutagenesis strategy have potential application in industrial fuel ethanol production from lignocellulosic hydrolysates