Translational diffusion, molecular brightness, and energy transfer analysis of mEGFP-linker-mScarlet-I crowding biosensor using fluorescence correlation spectroscopy.

Recently, we have investigated the sensitivity of an mEGFP-linker-mScarlet-I construct (GE2.3) in response to macromolecular crowding using ensemble time-resolved two-photon (2P) fluorescence measurements [Mersch , 2024, (5), 3927-3940] as a point of reference for developing a single-molecule approach for Förster resonance energy transfer (FRET). Here, we investigate the fluorescence fluctuations, FRET, molecular brightness, and translational diffusion of GE2.

Two-photon excited-state dynamics of mEGFP-linker-mScarlet-I crowding biosensor in controlled environments.

Macromolecular crowding affects many cellular processes such as diffusion, biochemical reaction kinetics, protein-protein interactions, and protein folding. Mapping the heterogeneous, dynamic crowding in living cells or tissues requires genetically encoded, site-specific, crowding sensors that are compatible with quantitative, noninvasive fluorescence micro-spectroscopy. Here, we carried out time-resolved 2P-fluorescence measurements of a new mEGFP-linker-mScarlet-I macromolecular crowding construct (GE2.

Efficient Chemical-Free Degradation of Waterborne Micropollutants with an Immobilized Dual-Porous TiO Photocatalyst.

Photocatalytic advanced oxidation processes (AOPs) promise a chemical-free route to energy-efficient degradation of waterborne micropollutants if long-standing mass transfer and light management issues can be overcome. Herein, we developed a dual-porous photocatalytic system consisting of a mesoporous (i.e.

Molecular Brightness Approach for FRET Analysis of Donor-Linker-Acceptor Constructs at the Single Molecule Level: A Concept.

In this report, we have developed a simple approach using single-detector fluorescence autocorrelation spectroscopy (FCS) to investigate the Förster resonance energy transfer (FRET) of genetically encoded, freely diffusing crTC2.1 (mTurquoise2.1-linker-mCitrine) at the single molecule level.

The Dockstore: enhancing a community platform for sharing reproducible and accessible computational protocols.

Dockstore (https://dockstore.org/) is an open source platform for publishing, sharing, and finding bioinformatics tools and workflows. The platform has facilitated large-scale biomedical research collaborations by using cloud technologies to increase the Findability, Accessibility, Interoperability and Reusability (FAIR) of computational resources, thereby promoting the reproducibility of complex bioinformatics analyses.

Fluorescence depolarization dynamics of ionic strength sensors using time-resolved anisotropy.

Eukaryotic cells exploit dynamic and compartmentalized ionic strength to impact a myriad of biological functions such as enzyme activities, protein-protein interactions, and catalytic functions. Herein, we investigated the fluorescence depolarization dynamics of recently developed ionic strength biosensors (mCerulean3-linker-mCitrine) in Hofmeister salt (KCl, NaCl, NaI, and NaSO) solutions. The mCerulean3-mCitrine acts as a Förster resonance energy transfer (FRET) pair, tethered together by two oppositely charged α-helices in the linker region.

Ocean acidification causes variable trait-shifts in a coral species.

High pCO habitats and their populations provide an unparalleled opportunity to assess how species may survive under future ocean acidification conditions, and help to reveal the traits that confer tolerance. Here we utilize a unique CO vent system to study the effects of exposure to elevated pCO2 on trait-shifts observed throughout natural populations of Astroides calycularis, an azooxanthellate scleractinian coral endemic to the Mediterranean. Unexpected shifts in skeletal and growth patterns were found.

FRET Analysis of Ionic Strength Sensors in the Hofmeister Series of Salt Solutions Using Fluorescence Lifetime Measurements.

Living cells are complex, crowded, and dynamic and continually respond to environmental and intracellular stimuli. They also have heterogeneous ionic strength with compartmentalized variations in both intracellular concentrations and types of ions. These challenges would benefit from the development of quantitative, noninvasive approaches for mapping the heterogeneous ionic strength fluctuations in living cells.

The right mindset: stress mindset moderates the association between perceived stress and depressive symptoms.

Having a stress-is-enhancing mindset - the extent to which one believes the effects of stress are enhancing rather than debilitating - is associated with greater health and well-being, and may mitigate negative outcomes associated with experiencing stress. The present study aimed to examine stress mindset and whether it moderates the association between perceived stress and mental health outcomes. Participants (= 293) completed questionnaires assessing their stress mindset, perceived stress level, and current depressive and anxiety symptoms.

Macromolecular crowding effects on energy transfer efficiency and donor-acceptor distance of hetero-FRET sensors using time-resolved fluorescence.

Living cells are crowded with macromolecules and organelles, which affect a myriad of biochemical processes. As a result, there is a need for sensitive molecular sensors for quantitative, site-specific assessment of macromolecular crowding. Here, we investigated the excited-state dynamics of recently developed hetero-FRET sensors (mCerulean3-linker-mCitrine) in homogeneous and heterogeneous environments using time-resolved fluorescence measurements, which are compatible with fluorescence lifetime imaging microscopy (FLIM).

Crowding Effects on Energy-Transfer Efficiencies of Hetero-FRET Probes As Measured Using Time-Resolved Fluorescence Anisotropy.

Macromolecular crowding is prevalent in all living cells due to the presence of large biomolecules and organelles. Cellular crowding is heterogeneous and is known to influence biomolecular transport, biochemical reactions, and protein folding. Emerging evidence suggests that some cell pathologies may be correlated with compartmentalized crowding.

Rotational and translational diffusion of size-dependent fluorescent probes in homogeneous and heterogeneous environments.

Living cells are crowded with dynamic distributions of macromolecules and organelles that influence protein diffusion, molecular transport, biochemical reactions, and protein assembly. Here, we test the hypothesis that the diffusion of single molecules deviates from Brownian motion as described by the Stokes-Einstein model in a manner that depends on the viscosity range, the chemical structure of both the diffusing species and the crowding agents, and the spatio-temporal resolution of the employed analytical methods. Our size-dependent fluorescent probes are rhodamine-110, quantum dots, enhanced green fluorescent proteins (EGFP), and mCerulean3-linker-mCitrine FRET probes with various linker length and flexibility.

Momentary experiential avoidance: Within-person correlates, antecedents, and consequences and between-person moderators.

We used ecological momentary assessment to investigate momentary correlates, antecedents, and consequences of experiential avoidance (EA), and to explore whether depression and anxiety moderate these within-person relationships. Participants recorded their mood, thoughts, stress, and EA four times daily for one week. Baseline depression and anxiety were associated with EA.

Direct Detection of Multiple Acidic Proton Sites in Zeolite HZSM-5.

Direct observation of multiple reactive sites in the zeolite HZSM-5, a member of the MFI family of zeolite structures, contradicts the traditional view of only one type of active protonic species in industrially important zeolites. In addition to the well-known Brönsted acid site proton, two other protonic species undergo room-temperature hydrogen-deuterium exchange with an alkane hydrocarbon reagent, including one zeolite moiety characterized by a broad H chemical shift at ca. 12-15 ppm that is reported here for the first time.

Early Transcriptional Responses during Heat Stress in the Coral Acropora hyacinthus.

Corals respond to heat pulses that cause bleaching with massive transcriptional change, but the immediate responses to stress that lead up to these shifts have never been detailed. Understanding these early signals could be important for identifying the regulatory mechanisms responsible for bleaching and how these mechanisms vary between more and less resilient corals. Using RNA sequencing (RNAseq) and sampling every 30 minutes during a short-term heat shock, we found that components of the transcriptome were significantly upregulated within 90 min and after a temperature increase of +2 °C.

Fluorescence Dynamics of a FRET Probe Designed for Crowding Studies.

Living cells are crowded with macromolecules and organelles. As a result, there is an urgent need for molecular sensors for quantitative, site-specific assessment of the macromolecular crowding effects on a myriad of biochemical processes toward quantitative cell biology and biophysics. Here we investigate the excited-state dynamics and translational diffusion of a novel FRET sensor (mCerulean-linker-mCitrine) in a buffer (PBS, pH 7.

N gas egress from patients' airways during LN spray cryotherapy.

Spray cryotherapy using liquid nitrogen (LN) is a general surgical tool used to ablate benign or malignant lesions. Adequate egress of the gaseous nitrogen (N) generated during this process must be provided for safe use when LN is used within the body rather than topically. When delivered to either the gastrointestinal tract (requiring active venting via a suction tube) or body cavities open to room barometric pressure (such as lung airways) allowing for passive venting, the N gas generated from the boiling process must be evacuated.

Metered Cryospray™: a novel uniform, controlled, and consistent in vivo application of liquid nitrogen cryogenic spray.

In this article, a novel cryotherapy approach using a uniform, controlled, and consistent in vivo application of liquid nitrogen (LN) spray as a Metered Cryospray (MCS) process is described. Although MCS may be used for many potential clinical applications, this paper focuses on the development that led to the controlled and consistent delivery of radial LN cryogen spray in order to generate a uniform circumferential effect and how the amount of MCS can be adapted to specifically ablate targeted diseases within a patient's lumen such as an airway or esophagus.

Investigating the widespread introduction of a tropical marine fouling species.

Little is known about the number and rate of introductions into terrestrial and marine tropical regions, and if introduction patterns and processes differ from temperate latitudes. Botryllid ascidians (marine invertebrate chordates) are an interesting group to study such introduction differences because several congeners have established populations across latitudes. While temperate botryllid invasions have been repeatedly highlighted, the global spread of tropical Botrylloides nigrum (Herdman, 1886) has been largely ignored.

Solution-based synthesis and purification of zinc tin phosphide nanowires.

The solution-based synthesis of nanoscale earth-abundant semiconductors has the potential to unlock simple, scalable, and tunable material processes which currently constrain development of novel compounds for alternative energy devices. One such promising semiconductor is zinc tin phosphide (ZnSnP2). We report the synthesis of ZnSnP2 nanowires via a solution-liquid-solid mechanism utilizing metallic zinc and tin in decomposing trioctylphosphine (TOP).