Picosecond Fluorescence Relaxation Spectroscopy of the Calcium-Discharged Photoproteins Aequorin and Obelin

Addition of calcium ions to the Ca2+-regulated photoproteins, such as aequorin and obelin, produces a blue bioluminescence originating from a fluorescence transition of the protein-bound product, coelenteramide. The kinetics of several transient fluorescent species of the bound coelenteramide is resolved after picosecond-laser excitation and streak camera detection. The initially formed spectral distributions at picosecond-times are broad, evidently comprised of two contributions, one at higher energy (25000 cm-1) assigned as from the Ca2+-discharged photoprotein-bound coelenteramide in its neutral state. This component decays much more rapidly (t1/2 2 ps) in the case of the Ca2+-discharged obelin than aequorin (t1/2 30 ps). The second component at lower energy shows several intermediates in the 150-500 ps times, with a final species having spectral maxima 19400 cm-1, bound to Ca2+-discharged obelin, and 21300 cm-1, bound to Ca2+-discharged aequorin, and both have a fluorescence decay lifetime of 4 ns. It is proposed that the rapid kinetics of these fluorescence transients on the picosecond time scale, correspond to times for relaxation of the protein structural environment of the binding cavit

Biosynthesis of Gold Nanoparticles by Foliar Broths: Roles of Biocompounds and Other Attributes of the Extracts

Biosynthesis of nanoparticles has arisen as a promising alternative to conventional synthetic methodologies owing to its eco-friendly advantages, and the involved bioprotocol still needs further clarification. This research, for the first time from the standpoint of statistics, confirmed an electrostatic force or ionic bond-based interaction between the chloroauric ions and the involved bioconstituents and manifested that reducing sugars and flavonoids were both important reductants responsible for conversion of Au(III) to Au(0). The result also demonstrated that the proteins were not the reducing agents, yet they might be protection agents in biosynthesis of gold nanoparticles (GNPs). Besides, a significant linear relationship was found between the anti-oxidant ability of the foliar broths and their capability to reduce Au(III) into Au(0). Furthermore, the preliminary investigation based on the boxplot on the size/shape distribution of the biosynthesized GNPs revealed that gold nanospheres with higher degree of homogeneity in size tended to be promoted by foliar broths containing higher content of reducing sugars/flavonoids and proteins. Otherwise, i.e., for those broths with lower content of the above biocompounds, sphere GNPs of wider size distribution or even gold nanotriangles tended to be fabricated

Disease characteristics of MCT8 deficiency : an international, retrospective, multicentre cohort study

Background Disordered thyroid hormone transport, due to mutations in the SLC16A2 gene encoding monocarboxylate transporter 8 (MCT8), is characterised by intellectual and motor disability resulting from cerebral hypothyroidism and chronic peripheral thyrotoxicosis. We sought to systematically assess the phenotypic characteristics and natural history of patients with MCT8 deficiency. Methods We did an international, multicentre, cohort study, analysing retrospective data from Jan 1, 2003, to Dec 31, 2019, from patients with MCT8 deficiency followed up in 47 hospitals in 22 countries globally. The key inclusion criterion was genetically confirmed MCT8 deficiency. There were no exclusion criteria. Our primary objective was to analyse the overall survival of patients with MCT8 deficiency and document causes of death. We also compared survival between patients who did or did not attain full head control by age 1·5 years and between patients who were or were not underweight by age 1–3 years (defined as a bodyweight-for-age Z score <–2 SDs or <5th percentile according to WHO definition). Other objectives were to assess neurocognitive function and outcomes, and clinical parameters including anthropometric characteristics, biochemical markers, and neuroimaging findings. Findings Between Oct 14, 2014, and Jan 17, 2020, we enrolled 151 patients with 73 different MCT8 (SLC16A2) mutations. Median age at diagnosis was 24·0 months (IQR 12·0-60·0, range 0·0-744·0). 32 (21%) of 151 patients died; the main causes of mortality in these patients were pulmonary infection (six [19%]) and sudden death (six [19%]). Median overall survival was 35·0 years (95% CI 8·3–61·7). Individuals who did not attain head control by age 1·5 years had an increased risk of death compared with patients who did attain head control (hazard ratio [HR] 3·46, 95% CI 1·76–8·34; log-rank test p=0·0041). Patients who were underweight during age 1–3 years had an increased risk for death compared with patients who were of normal bodyweight at this age (HR 4·71, 95% CI 1·26–17·58, p=0·021). The few motor and cognitive abilities of patients did not improve with age, as evidenced by the absence of significant correlations between biological age and scores on the Gross Motor Function Measure-88 and Bayley Scales of Infant Development III. Tri-iodothyronine concentrations were above the age-specific upper limit in 96 (95%) of 101 patients and free thyroxine concentrations were below the age-specific lower limit in 94 (89%) of 106 patients. 59 (71%) of 83 patients were underweight. 25 (53%) of 47 patients had elevated systolic blood pressure above the 90th percentile, 34 (76%) of 45 patients had premature atrial contractions, and 20 (31%) of 64 had resting tachycardia. The most consistent MRI finding was a global delay in myelination, which occurred in 13 (100%) of 13 patients. Interpretation Our description of characteristics of MCT8 deficiency in a large patient cohort reveals poor survival with a high prevalence of treatable underlying risk factors, and provides knowledge that might inform clinical management and future evaluation of therapies

Study of tetrabutylammonium bromide in aqueous solution by neutron scattering

The study of electrolyte solutions by neutron scattering is an example of the large range of possibilities of the technique. Structure and dynamics at different time and length scales, discrimination of global from local motions, separation of coherent from incoherent contributions are necessary to embrace the complexity of a subject where charge and hydrophobicity play important and competitive roles. The behaviour of aqueous solutions of tetrabutylammonium bromide is studied here by several neutron scattering techniques: Small Angle Neutron Scattering, Neutron Diffraction, Time-of-Flight and Neutron Spin Echo. We concentrate on the conformation and dynamics of the hydrophobic cations. In particular, the center-of-mass (CoM) motion of the cation at the microscopic scale is best described via the low Q coherent signal, as measured by Neutron Spin Echo. Due to a possible cage formation effect in the TBABr solution, at the scale of the distance between cations, the cationic CoM relaxation time is larger than that predicted by a simple extrapolation of results issued from the hydrodynamic regime and those obtained from the incoherent signal analysis

Diffusion of water in clays – microscopic simulation and neutron scattering

10 pagesThe dynamics of water in porous charged media (montmorillonite clay) is investigated on the picosecond-timescale by quasi-elastic neutron scattering (time-of-flight (TOF) and neutron spin echo (NSE) techniques) and classical molecular dynamics simulations. Correspondence is discussed not only in terms of integrated quantities such as diffusion coefficients but also more directly on the level of intermediate scattering functions. Both simulated and experimental water diffusion coefficients are of the order of 5–10 × 10-10 m2 s-1. Closer analysis suggests that, unlike NSE, TOF and simulation underestimate relaxation times in the low-Q region due to insufficiently large correlation times probed. Comparison between experimental and simulated dynamics is rendered difficult by the features of the real montmorillonite clay (interstratification, mesoporous water) omitted in the model. For the de-coupling of phenomena in a real clay, a more complete set of data for the montmorillonite clay (different ions, hydration states) or the use of other, in some respect more homogeneous clays (hectorite, vermiculite) is suggested

Dynamic study of N’N-dimethylparanitroaniline encapsulated in silicalite-1 matrix using neutron spin-echo spectroscopy

The present work focuses on the dynamic studies of N’N-dimethyl-paranitroaniline (dmpNA) encapsulated in silicalite zeolite. Quasielastic neutron scattering (QENS) experiments are carried out using neutron spin-echo technique. Polarisation of the scattered neutron beam is measured at carefully chosen values of Q = 0.35, 0.9, 1.1 and 1.45  Å−1 at fixed T = 298  K and at fixed Q = 0.9  Å−1 at 150, 200, 250 and 298 K. This gives insight into the motion and the related activation energy of the guest dmpNA molecule. The quasielastic signal observed in the present system within the time range considered is due to fast local rotational motions of protons of the end methyl groups. The results are in good agreement with the dynamics of methyl group rotations reported in the literature by back-scattering QENS technique

Experiments – Simulations – Theories: Multiscale approaches for solutions

The theory of complex media depends on various levels of description. At the macroscopic scale, hydrodynamic equations are used but they are not valid at small scales, the latter domain can be very important if slow processes occur in the system. At the microscopic scale, molecular dynamics describe the motion of the atoms individually. The two methods which correspond to different levels of description are complementary, but they are not the only ones. Intermediate methods based on continuous solvent model are able to take proper account of the solute dynamics and they can be applied at an intermediate time scale. We present such multi-scale descriptions in the case of solutions. Such an approach is all the more important in the context of of neutron science since neutron scattering experiments are now able to explore a large time domain: they are able to encompass the intermediate time domain for which intermediate methods are unavoidable