Search

The molecular energy transfer is crucial for many different physicochemical processes. The efficiency of traditional resonance energy transfer relies on dipole–dipole distance between molecules and becomes negligible when the distance is larger than ∼ 10 nm, which is difficult to overcome. Cavity polariton, formed when placing molecules inside the cavity, is a promising way to surmount the distance limit. By hybridizing a two-level atom (TLA) and a lithium fluoride (LiF) molecule with a cavity, we numerically simulate the reaction process and the energy transfer between them. Our results show that the TLA can induce a deep potential well, which can be seen as a replica of the potential energy surface of bare LiF, acting as a reservoir to absorb/release the molecular kinetic energy. In addition, the energy transfer shows a molecular nuclear kinetic energy dependent behavior, namely, more nuclear kinetic energy igniting more energy transfer. These findings show us a promising way to manipulate the energy transfer process within the cavity using an intentional TLA, which can also serve as a knob to control the reaction process. [ABSTRACT FROM AUTHOR]

Phycocyanin 620 (PC620) is the outermost light-harvesting complex in phycobilisome of cyanobacteria, engaged in light collection and energy transfer to the core antenna, allophycocyanin. Recently, long-lived exciton–vibrational coherences have been observed in allophycocyanin, accounting for the coherent energy transfer [Zhu et al., Nat. Commun. 15, 3171 (2024)]. PC620 has a nearly identical spatial location of three α84–β84 phycocyanobilin pigment pairs to those in allophycocyanin, inferring an existence of possible coherent energy transfer pathways. However, whether PC620 undergoes coherent or incoherent energy transfer remains debated. Furthermore, accurate determination of energy transfer rates in PC620 is still necessary owing to the spectral overlap and broadening in conventional time-resolved spectroscopic measurements. In this work, the energy transfer process within PC620 was directly resolved by polarization-controlled two dimensional electronic spectroscopy (2DES) and global analysis. The results show that the energy transfer from α84 to the adjacent β84 has a lifetime constant of 400 fs, from β155 to β84 of 6–8 ps, and from β155 to α84 of 66 ps, fully conforming to the Förster resonance energy transfer mechanism. The circular dichroism spectrum also reveals that the α84–β84 pigment pair does not form excitonic dimer, and the observed oscillatory signals are confirmed to be vibrational coherence, excluding the exciton–vibrational coupling. Nodal line slope analysis of 2DES further reveals that all the vibrational modes participate in the energy dissipation of the excited states. Our results consolidate that the ultrafast energy transfer process in PC620 is incoherent, where the twisted conformation of α84 is suggested as the main cause for preventing the formation of α84–β84 excitonic dimer in contrast to allophycocyanin. [ABSTRACT FROM AUTHOR]

It is increasingly clear that cellular metabolic function varies not just between cells of different tissues, but also within tissues and cell types. In this essay, we envision how differences in central carbon metabolism arise from multiple sources, including the cell cycle, circadian rhythms, intrinsic metabolic cycles, and others. We also discuss and compare methods that enable such variation to be detected, including single-cell metabolomics and RNA-sequencing. We pay particular attention to biosensors for AMPK and central carbon metabolites, which when used in combination with metabolic perturbations, provide clear evidence of cellular variance in metabolic function.

In the recently discovered proton-coupled energy transfer (PCEnT) mechanism, the transfer of electronic excitation energy between donor and acceptor chromophores is coupled to a proton transfer reaction. Herein, we develop a general theory for PCEnT and derive an analytical expression for the nonadiabatic PCEnT rate constant. This theory treats the transferring hydrogen nucleus quantum mechanically and describes the PCEnT process in terms of nonadiabatic transitions between reactant and product electron–proton vibronic states. The rate constant is expressed as a summation over these vibronic states, and the contribution of each pair of vibronic states depends on the square of the vibronic coupling as well as the spectral convolution integral, which can be viewed as a generalization of the Förster-type spectral overlap integral for vibronic rather than electronic states. The convolution integral also accounts for the common vibrational modes shared by the donor and acceptor chromophores for intramolecular PCEnT. We apply this theory to model systems to investigate the key features of PCEnT processes. The excited vibronic states can contribute significantly to the total PCEnT rate constant, and the common modes can either slow down or speed up the process. Because the pairs of vibronic states that contribute the most to the PCEnT rate constant may correspond to spectroscopically dark states, PCEnT could occur even when there is no apparent overlap between the donor emission and acceptor absorption spectra. This theory will assist in the interpretation of experimental data and will guide the design of additional PCEnT systems. [ABSTRACT FROM AUTHOR]

Electronic energy transfer in molecular wires is usually theoretically investigated with a harmonic bath to model the environment. The present study is a continuation of our previous work [A. Sindhu and A. Jain, Chem. Phys. Chem. 23, e2022003 (2022)] on studying the dynamics of molecular wires using surface hopping simulations. We extend our study to a 7-site model Hamiltonian and investigate the effects of an anharmonic bath on coherent energy transfer in molecular wires. We show that oscillatory and coherent population dynamics remain intact even in the presence of the anharmonic bath and further highlight the multiple channels available for energy flow in molecular wires. [ABSTRACT FROM AUTHOR]

A previously developed semiclassical theory of nonadiabatic energy transfer is used to analyze electronic excitation and quenching in three-dimensional atomic collisions. The predicted transition probabilities, cross sections, and rate coefficients are compared with the quantum scattering calculations for O + O and N + N, for the same interaction potentials and nonadiabatic coupling, and with the experimental data where available. The theory predictions are in very good agreement with quantum scattering, at the conditions when the energy transfer is dominated by a single pair of adiabatic potentials. Closed-form analytic expressions for the cross sections and rate coefficients are obtained, for both the strongly and weakly coupled cases. The results quantify and illustrate the effect of the interaction potentials and their coupling on the energy transfer. The analytic cross sections and rate coefficients are in good agreement with the numerical predictions. The same approach has been used to predict the rate coefficients of electronic excitation and quenching in collisions of N + O atoms. The fidelity of these predictions may be improved considerably if accurate potentials for the excited electronic states of N + O and their coupling are available. The applicability of the semiclassical theory for the prediction of the rates of heavy particle impact excitation in atom–molecule collisions is discussed. [ABSTRACT FROM AUTHOR]

Nonmuscle myosin II (NMII) generates cytoskeletal forces that drive cell division, embryogenesis, muscle contraction and many other cellular functions. However, at present there is no method that can directly measure the forces generated by myosins in living cells. Here, we describe a Förster resonance energy transfer (FRET)-based tension sensor that can detect myosin-associated force along the filamentous actin network. Fluorescence lifetime imaging microscopy (FLIM)-FRET measurements indicate that the forces generated by NMII isoform B (NMIIB) exhibit significant spatial and temporal heterogeneity as a function of donor lifetime and fluorophore energy exchange. These measurements provide a proxy for inferred forces that vary widely along the actin cytoskeleton. This initial report highlights the potential utility of myosin-based tension sensors in elucidating the roles of cytoskeletal contractility in a wide variety of contexts.

N-methyl-D-aspartate receptors are ionotropic glutamate receptors that mediate synaptic transmission and plasticity. Variable GluN2 subunits in diheterotetrameric receptors with identical GluN1 subunits set very different functional properties. To understand this diversity, we use single-molecule fluorescence resonance energy transfer (smFRET) to measure the conformations of the ligand binding domain and modulatory amino-terminal domain of the common GluN1 subunit in receptors with different GluN2 subunits. Our results demonstrate a strong influence of the GluN2 subunits on GluN1 rearrangements, both in non-agonized and partially agonized activation intermediates, which have been elusive to structural analysis, and in the fully liganded state. Chimeric analysis reveals structural determinants that contribute to these subtype differences. Our study provides a framework for understanding the conformational landscape that supports highly divergent levels of activity, desensitization, and agonist potency in receptors with different GluN2s and could open avenues for the development of subtype-specific modulators.

The compound 2,3-diazabicyclo[2.2.2]oct-2-ene (DBO) is a versatile fluorophore widely used in Förster resonance energy transfer (FRET) spectroscopy studies due to its remarkable sensitivity, enabling precise donor–acceptor distance measurements, even for short peptides. Integrating time-resolved and FRET spectroscopies with molecular dynamics simulations provides a robust approach to unravel the structure and dynamics of biopolymers in a solution. This study investigates the structural behavior of three octapeptide variants: Trp-(Gly-Ser)3-Dbo, Trp-(GlyGly)3-Dbo, and Trp-(SerSer)3-Dbo, where Dbo represents the DBO-containing modified aspartic acid, using molecular dynamics simulations. Glycine- and serine-rich amino acid fragments, common in flexible protein regions, play essential roles in functional properties. Results show excellent agreement between end-to-end distances, orientational factors from simulations, and the available experimental and theoretical data, validating the reliability of the GROMOS force field model. The end-to-end distribution, modeled using three Gaussian distributions, reveals a complex shape, confirmed by cluster analysis highlighting a limited number of significant conformations dominating the peptide landscape. All peptides predominantly adopt a disordered state in the solvent, yet exhibit a compact shape, aligning with the model of disordered polypeptide chains in poor solvents. Conformations show marginal dependence on chain composition, with Ser-only chains exhibiting slightly more elongation. This study enhances our understanding of peptide behavior, providing valuable insights into their structural dynamics in solution. [ABSTRACT FROM AUTHOR]

Conical intersection (CI) leads to fast electronic energy transfer. However, Hamm and Stock [Phys. Rev. Lett. 109, 173201 (2012)] showed the existence of a vibrational CI and its role in vibrational energy relaxation. In this paper, we further investigate the vibrational energy relaxation using an isolated model Hamiltonian system of four vibrational modes with two distinctively different timescales (two fast modes and two slow modes). We show that the excitation of the slow modes plays a crucial role in the energy relaxation mechanism. We also analyze the system from a mixed quantum-classical (surface hopping method) and a completely classical point of view. Notably, surface hopping and even classical simulations also capture fast energy relaxation, which is a signature of CI's existence. [ABSTRACT FROM AUTHOR]

The structural diversity of biological macromolecules in different environments contributes complexity to enzymological processes vital for cellular functions. Fluorescence resonance energy transfer and electron microscopy are used to investigate the enzymatic reaction of T4 DNA ligase catalyzing the ligation of nicked DNA. The data show that both the ligase-AMP complex and the ligase-AMP-DNA complex can have four conformations. This finding suggests the parallel occurrence of four ligation reaction pathways, each characterized by specific conformations of the ligase-AMP complex that persist in the ligase-AMP-DNA complex. Notably, these complexes have DNA bending angles of ≈0°, 20°, 60°, or 100°. The mechanism of parallel reactions challenges the conventional notion of simple sequential reaction steps occurring among multiple conformations. The results provide insights into the dynamic conformational changes and the versatile attributes of T4 DNA ligase and suggest that the parallel multiple reaction pathways may correspond to diverse T4 DNA ligase functions. This mechanism may potentially have evolved as an adaptive strategy across evolutionary history to navigate complex environments.

BackgroundChronic sympathetic stimulation drives desensitization and downregulation of β1 adrenergic receptor (β1AR) in heart failure. We aim to explore the differential downregulation subcellular pools of β1AR signaling in the heart.Methods and resultsWe applied chronic infusion of isoproterenol to induced cardiomyopathy in male C57BL/6J mice. We applied confocal and proximity ligation assay to examine β1AR association with L-type calcium channel, ryanodine receptor 2, and SERCA2a ((Sarco)endoplasmic reticulum calcium ATPase 2a) and Förster resonance energy transfer-based biosensors to probe subcellular β1AR-PKA (protein kinase A) signaling in ventricular myocytes. Chronic infusion of isoproterenol led to reduced β1AR protein levels, receptor association with L-type calcium channel and ryanodine receptor 2 measured by proximity ligation (puncta/cell, 29.65 saline versus 14.17 isoproterenol, P

Detecting ascorbate imbalances is crucial for timely disease diagnosis and sample monitoring across various conditions. In this study, we synthesized N,S-GQDs and employed the fluorescence quenching effect of CoOOH nanosheets through resonance energy transfer (FRET) to visually detect changes in ascorbate levels. Upon the addition of ascorbic acid (AA), the reduction reaction with CoOOH induced fluorescence recovery in N,S-GQDs. Our findings demonstrate that the synthesized N,S-GQDs/CoOOH nanocomplexes provide a reliable probe for fluorescence-based detection of ascorbic acid within a linear range of 2–82 μM, with a detection limit of 48.3 nM. Additionally, quantitative determination of ascorbic acid in blood and orange juice samples was successfully achieved using the N,S-GQDs/CoOOH, with good recovery rates. Importantly, confocal fluorescence imaging experiments conducted on MCF-7 cells and zebrafish confirmed that the synthetic N,S-GQDs/CoOOH can effectively visualize the overexpression of ascorbic acid in vivo. [ABSTRACT FROM AUTHOR]

The behaviour of photoexcited electrons in light-responsive coordination polymers (CPs) significantly determines their performance in fluorescence sensing, smart materials, photoelectric display and photocatalysis, and this behaviour can be skilfully manipulated by optimizing the geometric and electronic structures of the ligand field around the metal ion. To reveal the micro-environmental effect of the ligand field on the bandgap and photoexcited electrons, three semiconductive Cd(II)-based CPs have been achieved through coordination of π-conjugated 2,6-bis(2-pyrazin-2-yl)-4-(4-(tetrazol-5-yl)phenyl)pyridine (HL) and 1,3,5-benzenetricarboxylic acid (H3BTC) with Cd(II) ion. Possessing N3O4 and O6 donors, complex 1 assumed an extended triple-decker sandwich structure with a central {Cd2(BTC)} sheet terminated up and down by planar L− spacers. Two quadrangular {Cd2(L)2} dimers with N4O3 and N4O2 donors were propagated by doubly deprotonated HBTC2− connectors to the cationic (4 4) layer of 2. The reproducible {Cd2(L)2} dimers with only N4O3 donors in 3 were extended by mirror-symmetric HBTC2− linkers to a bent chain. The distinct donor combinations in the local ligand fields of Cd(II) ions dominated the bandgaps and movements of the charge-carriers. Emitting intense steel blue fluorescence, complex 1 served as a highly efficient bilirubin probe with a quenching constant up to 9.48 × 104 M−1 driven by photo-induced electron charge and Förster resonance energy transfer. In contrast, complex 2, with excellent separation efficiency of charge carriers, acted as a photocatalyst to completely degrade methylene blue up to 97% within 90 minutes of UV irradiation. Apparently, slight modifications of the surroundings of the ligand field by a heterodonor strategy achieved a remarkable effect on the photophysical properties of the semiconductive CPs, providing valuable hints for the construction of photosensitive CPs. [ABSTRACT FROM AUTHOR]

Microbial rhodopsin-derived genetically encoded voltage indicators (GEVIs) are powerful tools for mapping bioelectrical dynamics in cell culture and in live animals. Förster resonance energy transfer (FRET)-opsin GEVIs use voltage-dependent quenching of an attached fluorophore, achieving high brightness, speed, and voltage sensitivity. However, the voltage sensitivity of most FRET-opsin GEVIs has been reported to decrease or vanish under two-photon (2P) excitation. Here, we investigated the photophysics of the FRET-opsin GEVIs Voltron1 and Voltron2. We found that the previously reported negative-going voltage sensitivities of both GEVIs came from photocycle intermediates, not from the opsin ground states. The voltage sensitivities of both GEVIs were nonlinear functions of illumination intensity; for Voltron1, the sensitivity reversed the sign under low-intensity illumination. Using photocycle-optimized 2P illumination protocols, we demonstrate 2P voltage imaging with Voltron2 in the barrel cortex of a live mouse. These results open the door to high-speed 2P voltage imaging of FRET-opsin GEVIs in vivo. [ABSTRACT FROM AUTHOR]

Polymersomes are polymeric vesicles composed of amphiphilic block copolymers with a hydrophilic inner lumen and a hydrophobic membrane, capable of encapsulating both hydrophilic payloads, and hydrophobic payloads. The one‐pot synthesis and self‐assembly of polyethylene‐glycol‐based amphiphilic polythiourethanes consisting of a pendant methyl or aromatic ester are reported here. During the ring opening of cyclodithiocarbonate using ethylene diamine or hexyl diamine, generated thiols are utilized to modify the backbone with methyl, benzyl, and diphenyl esters. The aromatic pendant containing P3 and P4 has higher hydrophobic dye loading than the other methyl ester containing polymers P1 and P2. All the polymers have hydrodynamic diameters in the range of 40–98 nm. Atomic force microscopy (AFM) results show that all the polymers are able to form spherical aggregates with diameters of 38.1 ± 7.7 and 85.1 ± 27.4 nm. Among them, aromatic side‐chain‐containing polymers have 42% and 57% calcein encapsulation efficiency, whereas methyl‐ester‐containing polymers have 70% and 60% efficiency. The polymers with longer spacers (P2–P4) and aromatic groups (P3 and P4) have shown better stability and slow dye exchange over 48 h compared to the polymer with an aliphatic side chain and a shorter spacer probed by fluorescence resonance energy transfer (FRET) study. [ABSTRACT FROM AUTHOR]

Ribonucleoprotein (RNP) condensates partition RNA and protein into multiple liquid phases. The multiphasic feature of condensate-enriched components creates experimental challenges for distinguishing membraneless condensate functions from the surrounding dilute phase. We combined fluorescence lifetime imaging microscopy (FLIM) with phasor plot filtering and segmentation to resolve condensates from the dilute phase. Condensate-specific lifetimes were used to track protein-protein interactions by measuring FLIM-Förster resonance energy transfer (FRET). We used condensate FLIM-FRET to evaluate whether mRNA decapping complex subunits can form decapping-competent interactions within P-bodies. Condensate FLIMFRET revealed the presence of core subunit interactions within P-bodies under basal conditions and the disruption of interactions between the decapping enzyme (Dcp2) and a critical cofactor (Dcp1A) during oxidative stress. Our results show a context-dependent plasticity of the P-body interaction network, which can be rewired within minutes in response to stimuli. Together, our FLIM-based approaches provide investigators with an automated and rigorous method to uncover and track essential protein-protein interaction dynamics within RNP condensates in live cells. [ABSTRACT FROM AUTHOR]

The glucagon‐like peptide‐1 receptor (GLP‐1R) plays an important role in regulating insulin secretion and reducing body weight, making it a prominent target in the treatment of type 2 diabetes and obesity. Extensive research on GLP‐1R signaling has provided insights into the connection between receptor function and physiological outcomes, such as the correlation between Gs signaling and insulin secretion, yet the exact mechanisms regulating signaling remain unclear. Here, we explore the internalization pathway of GLP‐1R, which is crucial for controlling insulin release and maintaining pancreatic beta‐cell function. Utilizing a reliable and sensitive time‐resolved fluorescence resonance energy transfer (TR‐FRET) internalization assay, combined with HEK293‐derived knockout cell lines, we were able to directly compare the involvement of different endocytic machinery in GLP‐1R internalization. Our findings indicate that the receptor internalizes independently of arrestin and is dependent on Gs and Gi/o activation and G protein–coupled receptor kinase phosphorylation. Mechanistically, we observed that the receptor undergoes distinct clathrin‐ and caveolae‐mediated internalization in HEK293 cells. This study also investigated the role of arrestins in GLP‐1R function and regulation. These insights into key endocytic components that are involved in the GLP‐1R internalization pathway could enhance the rational design of GLP‐1R therapeutics for type 2 diabetes and other GLP‐1R‐related diseases. [ABSTRACT FROM AUTHOR]

Fusarium graminearum FgSte2 and FgSte3 are G-protein-coupled receptors (GPCRs) shown to play roles in hyphal chemotropism and fungal plant pathogenesis in response to activity arising from host-secreted peroxidases. Here, we follow up on the observation that chemotropism is dependent on both FgSte2 and FgSte3 being present; testing the possibility that this might be due to formation of an FgSte2-FgSte3 heterodimer. Bioluminescence resonance energy transfer (BRET) analyses were conducted in Saccharomyces cerevisiae, where the addition of horse radish peroxidase (HRP) was found to increase the transfer of energy from the inducibly expressed FgSte3-Nano luciferase donor, to the constitutively expressed FgSte2-yellow fluorescent protein (YFP) acceptor, compared to controls. A partial response was also detected when an HRP-derived ligand-containing extract was enriched from F. graminearum spores and applied instead of HRP. In contrast, substitution with pheromones or an unrelated bovine GPCR, rhodopsin-YFP used as acceptor, eliminated all BRET responses. Interaction results were validated by affinity pulldown and receptor expression was validated by confocal immunofluorescence microscopy. Taken together these findings demonstrate the formation of HRP and HRP-derived ligand stimulated heterodimers between FgSte2 and FgSte3. Outcomes are discussed from the context of the roles of ligands and reactive oxygen species in GPCR dimerization. [ABSTRACT FROM AUTHOR]

A dual-signal aptamer-based assay utilizing colorimetric and fluorescence techniques was developed for the determination of zearalenone (ZEN). The CdTe quantum dots, serving as the fluorescent signal source, were surface-modified onto Fe3O4@SiO2 and subsequently functionalized with the aptamer. The COF-Au was modified with complementary chain, which possessed peroxide (POD)-like enzyme properties, and could catalyze the peroxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to ox TMB, resulting in the generation of colorimetric signals. The two parts were merged based on the principle of base complementary pairing, resulting in an assembled structure exhibiting a diminished fluorescence signal due to the Förster resonance energy transfer (FRET) effect. Due to the higher affinity of the aptamer towards the target, the presence of ZEN resulted in the detachment of COF-Au, leading to an increase in supernatant concentration of COF-Au proportional to ZEN concentration. Consequently, this enhanced the catalytic ability and amplified the colorimetric signal. The fluorescence of precipitation increased simultaneously with the reduction of FRET, enabling linear detection of colorimetry in the range 0.5 ~ 10,000 μg·kg−1 and fluorescence in the range 0.1 ~ 10,000 μg·kg−1, with respective detection limits of 0.36 μg·kg−1 and 0.09 μg·kg−1. The spike recovery in wheat flour and corn ranged from 93.4 to 122.0%. This technology was simple to operate and had low cost and good application prospects. [ABSTRACT FROM AUTHOR]

Multi-wavelength emission fluorescent manganese-nitrogen co-doped carbon dots (Mn, N co-doped CDs) were synthesized by solvothermal method using β-cyclodextrin, O-phenylenediamine, and manganese chloride as raw materials. The prepared Mn, N co-doped CDs were used as fluorescent nanosensing platforms for the detection of metal ions and biomolecules and were found to be capable of fluorescence detection of tannic acid (TA) and hafnium (Hf) ion at 320, 380, and 480 nm excitation wavelengths with multi-response linear ranges of 0.7 ~ 1.2 µM and 6.35 ~ 13 µM and detection limits of 0.45 µM and 6.3 µM, respectively. The wide linear ranges and low detection limits may be due to the fluorescence resonance energy transfer effect between the platform and TA and Hf ions. In addition, it was found that Mn, N co-doped CDs had good photostability, biocompatibility, and low cytotoxicity, which could be used for in vitro fluorescence imaging of exogenous TA and Hf ion imaging in U2OS osteosarcoma cells. Thus, the probe has a promising application in biomedical fields as a new multi-responsive fluorescence nanosensing platform member. [ABSTRACT FROM AUTHOR]

Titin is the third contractile filament in the sarcomere, and it plays a critical role in sarcomere integrity and both passive and active tension. Unlike the thick and thin filaments, which are polymers of myosin and actin, respectively, titin is a single protein that spans from Z-disk to M-line. The N2A region within titin has been identified as a signaling hub for the muscle and is shown to be involved in multiple interactions. The insertion sequence (UN2A) within the N2A region was predicted as a potential binding site for the Ca2+-binding protein, S100A1. We demonstrate using a combination of size exclusion chromatography, surface plasmon resonance, and fluorescence resonance energy transfer that S100A1 can bind to the UN2A region. We further demonstrate that this interaction occurs under conditions where calcium is bound to S100A1, suggesting that the conformational shift in S100A1 when calcium binds is important. We also observed a conformational change in UN2A induced by shifts in pH, suggesting that conformational flexibility in UN2A plays a critical role in the interaction with S100A1. These results lead us to propose that the interaction of S100A1 and UN2A might act as a sensor to regulate titin’s function in response to physiological changes in the muscle. [ABSTRACT FROM AUTHOR]

Developing activatable photodynamic agents is becoming a promising mode for realizing selective photodynamic therapy (PDT) in cancer treatment. However, till now only a few H2S‐activatable photodynamic agents have been involved in this field. Here, we fabricated H2S‐activatable nano‐assemblies (P@TPPCY) for the treatment of colon cancer containing high concentration H2S. The H2S‐activatable photosensitizer (TPPCY) was synthesized by covalent conjugation between tetraphenylporphyrin (TPP) and heptamethine cyanine (Cy7), and then TPPCY was encapsulated by an amphiphilic polymer DSPE‐mPEG to form P@TPPCY nano‐assemblies. We demonstrated that the photosensitizing effect of TPP was efficiently quenched by Cy7 with strong absorption in the NIR region via fluorescence resonance energy transfer (FRET) effect. Interestingly, the FRET effect between Cy7 and TPP was attenuated by the decrease of absorption of Cy7 in the NIR region after the Cy7 reacted with H2S, and then the photosensitizing effect of TPP in P@TPPCY was activated. Strikingly, the P@TPPCY showed efficient H2S‐activatable photodynamic therapy (PDT) in vitro against HCT116 cells (human colon carcinoma cell line), thus this work provides a new method for realizing accurate treatment of colon cancer in virtue of high H2S concentration microenvironment. [ABSTRACT FROM AUTHOR]

Hyperphosphorylation and aggregation of MAPT (microtubule-associated protein tau) is a pathogenic hallmark of tauopathies and a defining feature of Alzheimer disease (AD). Pathological MAPT/tau is targeted by macroautophagy/autophagy for clearance after being sequestered within autophagosomes, but autophagy dysfunction is indicated in tauopathy. While mitochondrial bioenergetic deficits have been shown to precede MAPT/tau pathology in tauopathy brains, it is unclear whether energy metabolism deficiency is involved in the pathogenesis of autophagy defects. Here, we reveal that stimulation of anaplerotic metabolism restores defective oxidative phosphorylation (OXPHOS) in tauopathy neurons which, strikingly, leads to pronounced MAPT/tau clearance by boosting autophagy functionality through enhancements of mitochondrial biosynthesis and supply of phosphatidylethanolamine for autophagosome biogenesis. Furthermore, early anaplerotic stimulation of OXPHOS elevates autophagy activity and attenuates MAPT/tau pathology, thereby counteracting memory impairment in tauopathy mice. Taken together, our study sheds light on a pivotal role of mitochondrial bioenergetic deficiency in tauopathy-related autophagy defects and suggests a new therapeutic strategy to prevent the buildup of pathological MAPT/tau in AD and other tauopathy diseases. Abbreviation: AA: antimycin A; AD, Alzheimer disease; ATP, adenosine triphosphate; AV, autophagosome/autophagic vacuole; AZ, active zone; Baf-A1: bafilomycin A1; CHX, cycloheximide; COX, cytochrome c oxidase; DIV, days in vitro; DRG, dorsal root ganglion; ETN, ethanolamine; FRET, Förster/fluorescence resonance energy transfer; FTD, frontotemporal dementia; Gln, glutamine; HA: hydroxylamine; HsMAPT/Tau, human MAPT; IMM, inner mitochondrial membrane; LAMP1, lysosomal-associated membrane protein 1; LIs, lysosomal inhibitors; MDAV, mitochondria-derived autophagic vacuole; MmMAPT/Tau, murine MAPT; NFT, neurofibrillary tangle; OCR, oxygen consumption rate; Omy: oligomycin; OXPHOS, oxidative phosphorylation; PPARGC1A/PGC-1alpha: peroxisome proliferative activated receptor, gamma, coactivator 1 alpha; PE, phosphatidylethanolamine; phospho-MAPT/tau, hyperphosphorylated MAPT; PS, phosphatidylserine; PISD, phosphatidylserine decarboxylase;SQSTM1/p62, sequestosome 1; STX1, syntaxin 1; SYP, synaptophysin; Tg, transgenic; TCA, tricarboxylic acid; TEM, transmission electron microscopy. [ABSTRACT FROM AUTHOR]

Background: The unavoidable propagation of the coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has underscored the urgent requirement for efficacious therapeutic agents. The dried fruit of Terminalia chebula Retz., namely Chebulae Fructus, is widely used for treating bacterial and viral infectious diseases, which was witnessed to perform anti-SARS-CoV-2 activity in recommended Chinese patent medicine. Aim: SARS-CoV-2 main protease (Mpro) and papain-like protease (PLpro) present essential effects on SARS-CoV-2 replication and transcription, considering as the attractive targets for therapeutic intervention. In this study, we focused on the dual-target to obtain broad-spectrum antiviral candidates from Chebulae Fructus. Methods: The identified compounds from Chebulae Fructus were used to build a library in a previous study, which were evaluated by molecular docking to screen potential antiviral agents. The SARS-CoV-2 Mpro and PLpro were expressed in E. coli cells and purified. Fluorescence resonance energy transfer (FRET) and surface plasmon resonance (SPR) were utilized to verify the affinity with dual targets. SARS-CoV-2 wild-type, Omicron BA.5 and Omicron EG.5 variants were employed to validate their antiviral activities in vitro. Molecular dynamics simulation was conducted via Gromacs 2022 software in 500 ns to unveil the conformation stability. Results: Targeting on Mpro and PLpro, eight compounds were screened as the potential dual-target inhibitors in molecular docking. In FRET and SPR assays, 1,2,3,4,6-penta- O -galloyl- β -D-glucose (PGG) and 1,2,3,6-tetra- O -galloyl- β -D-glucose (TGG) showed good inhibitory activities with IC50 values ranging from 1.33 to 27.37 μM, and affinity with KD values ranging from 0.442 to 0.776 μM. Satisfactorily, both PGG and TGG display antiviral activity in vitro with EC50 values ranging from 3.20 to 37.29 μM, suggesting as the promising candidates against SARS-CoV-2. In molecular dynamics simulation study, the complexes of Mpro-PGG, Mpro-TGG, PLpro-PGG, and PLpro-TGG exhibited stability over 500 ns period, unveiling the potential interactions. Conclusion: PGG and TGG are the promising dual-target inhibitors of SARS-CoV-2, which may avoid drug resistance and have a good development prospect. The outcomes of this study provide an effective strategy to systematically explore the antiviral bioactive compounds from Chebulae Fructus. [ABSTRACT FROM AUTHOR]

This manuscript details the application of Isothermal Titration Calorimetry (ITC) to characterize the kinetics of 3CLpro, the main protease from the Severe Acute Respiratory Syndrome CoronaVirus-2 (SARS-CoV-2), and its inhibition by Ensitrelvir, a known non-covalent inhibitor. 3CLpro is essential for producing the proteins necessary for viral infection, which led to the COVID-19 pandemic. The ITC-based assay provided rapid and reliable measurements of 3CLpro activity, allowing for the direct derivation of the kinetic enzymatic constants KM and kcat by monitoring the thermal power required to maintain a constant temperature as the substrate is consumed. The manuscript highlights several advantages of the proposed ITC-based assay over traditional methods used to study 3CLpro, such as Förster Resonance Energy Transfer (FRET) and Liquid Chromatography-Mass Spectrometry (LC–MS) and overcomes the need for non-biological substrates or discontinuous post-reaction steps. The ease of application of the ITC method allowed for the determination of the temperature dependence of the catalytic constants, enabling the estimation of the reaction activation energy. Additionally, the assay was used to determine the inhibition mode and kinetic parameters for 3CLpro inhibition by Ensitrelvir. This molecule was revealed to act as a slow- and tight-binding inhibitor that forms an initial E•I complex (KI = 9.9 ± 0.7 nM) quickly transitioning to a tighter E•I* assembly (KI* = 1.1 ± 0.2 nM). This versatile calorimetric method is proposed for general use in the discovery and development of drugs targeting 3CLpro. [ABSTRACT FROM AUTHOR]

Fluorescence resonance energy transfer (FRET)‐based biosensors are powerful tools for studying second messengers with high temporal and spatial resolution. FRET is commonly detected by ratio imaging, but fluorescence lifetime imaging microscopy (FLIM), which measures the donor fluorophore's lifetime, offers a robust and more quantitative alternative. We have introduced and optimized four generations of FRET sensors for cAMP, based on the effector molecule Epac1, including variants for either ratio imaging or FLIM detection. Recently, Massengill and colleagues introduced additional mutations that improve cytosolic localization in these sensors, focusing on constructs optimized for ratio imaging. Here we present and briefly characterize these mutations in our dedicated FLIM sensors, finding they enhance cytosolic localization while maintaining performance comparable to original constructs. [ABSTRACT FROM AUTHOR]

The concentration of zinc ions in semen is significantly correlated with sperm viability and male fertility. In this work, a reliable ratiometric fluorescence probe (ZIF-9-TCPP) based on the efficient Förster resonance energy transfer (FRET) process between two luminophores, benzimidazole (BIM) and meso-tetra (4-carboxyphenyl) porphyrin (TCPP) for Zn2+ detection has been constructed, where the emissions of BIM and TCPP are used as reference and detection signals. The proximity of BIM and TCPP in one framework (ZIF-9-TCPP) and the overlapped spectra between BIM and TCPP afford the attainment of a highly efficient FRET (around 90% efficiency). Efficient FRET improves the fluorescence intensity of porphyrin to enhance the sensitivity of detection. The unique spectral shift resulting from Zn2+ binding to the porphyrin ring ensures the selectivity of detection. In addition, the response mechanism of the proposed ratiometric probes to Zn2+ has been investigated. This work provides a convenient way to design an efficient FRET system and a promising method for sperm mobility detection. [ABSTRACT FROM AUTHOR]

Progress in synthetic biology and nanotechnology plays at present a major role in the fabrication of sophisticated and miniaturized analytical devices that provide the means to tackle the need for new tools and methods for environmental and food safety. Significant research efforts have led to biosensing experiments experiencing a remarkable growth with the development and application of recombinant luminescent proteins (RLPs) being at the core of this boost. Integrating RLPs into biosensors has resulted in highly versatile detection platforms. These platforms include luminescent enzyme-linked immunosorbent assays (ELISAs), bioluminescence resonance energy transfer (BRET)–based sensors, and genetically encoded luminescent biosensors. Increased signal-to-noise ratios, rapid response times, and the ability to monitor dynamic biological processes in live cells are advantages inherent to the approaches mentioned above. Furthermore, novel fusion proteins and optimized expression systems to improve their stability, brightness, and spectral properties have enhanced the performance and pertinence of luminescent biosensors in diverse fields. This review highlights recent progress in RLP-based biosensing, showcasing their implementation for monitoring different contaminants commonly found in food and environmental samples. Future perspectives and potential challenges in these two areas of interest are also addressed, providing a comprehensive overview of the current state and a forecast of the biosensing strategies using recombinant luminescent proteins to come. [ABSTRACT FROM AUTHOR]

A BRET system is described, in which Nanoluciferase was fused to the lipid transfer protein CERT for efficient energy transfer to a Nile red‐labeled ceramide, which is either directly bound to CERT or transported to the adjacent Golgi membrane. Bulk formation of sphingomyelin, a major plasma membrane component in mammals, is dependent on CERT‐mediated transfer of its predecessor ceramide. CERT is considered a promising drug target but no direct cell‐based methods exist to efficiently identify inhibitors. The utility of the method was demonstrated by a library of 140 derivatives of the CERT inhibitor HPA‐12. These were obtained in a combinatorial synthesis using solid‐phase transacylation. Screening of the library led to six compounds that were picked and confirmed to be superior to HPA‐12 in a subsequent dose‐response study and also in an orthogonal lipidomics analysis. [ABSTRACT FROM AUTHOR]

A remarkable and unexpected increase in the photopolymerization efficiency of an acrylic resin by a bisacylphosphine oxide photoinitiator was observed when an optical brightener was present in the medium. High values for the maximal rates of photopolymerization were obtained by RT‐FTIR at 365 nm under a very low irradiance of 1 mW/cm2. Fluorescence studies revealed that the quenching process occurs through singlet‐singlet energy transfer between the first singlet excited state of the optical brightener and the ground state photoinitiator. This mechanism acts as an additional pathway for the excitation of the photoinitiator, thereby increasing the total amount of initiating radicals. Using the Förster resonance energy transfer model, we calculated the relative efficiency of the photosensitization process with respect to the direct excitation efficiency of the photoinitiator. The results demonstrate that the photosensitization process can be predicted, paving the way for further improvements in photoinitiating systems. [ABSTRACT FROM AUTHOR]

We have synthesized two dyads (dyad 1 and 2) comprising of tetraarylpyrrolo[3,2‐b]pyrrole (TAPP) and BODIPY. In dyad 1, two BODIPYs are directly connected with TAPP moiety whereas in dyad 2, BODIPYs are connected through phenylethynyl linkers. TAPP is a blue energy donor which is easy to synthesize and functionalize as compared to other well‐known blue energy donors like pyrene, perylene etc. This is the first report of using TAPP as an energy donor in BODIPY based dyad molecules. Complete quenching of TAPP fluorescence in the dyads suggests fast energy transfer from TAPP to BODIPY unit (ETE~99.9 %). Ultrafast fluorescence and transient absorption spectroscopic studies of dyad 1 showed TAPP to BODIPY energy transfer in 125 fs (kET=8.0×1012 s−1) which is one of the fastest energy transfer events in BODIPY based dyad reported so far. Whereas, in dyad 2, energy transfer is almost four times slower (480 fs, kET=2.1×1012 s−1). These results were rationalized by theoretical Förster formulations. This study shows that suitably matched optical properties of TAPP and BODIPY dyes along with their easy syntheses will be the key to develop highly efficient energy transfer systems in future for multiple applications. [ABSTRACT FROM AUTHOR]

Circularly polarized luminescence (CPL) materials offer exciting potential due to their unique light properties. However, controlling the color of afterglow emission, a key feature for many applications remains a challenge. Here, a co‐doping strategy is presented for precise control of multicolor CPL afterglow in films. This approach utilizes chiral naphthylphosphoric acid derivatives as energy and chirality transfer donors, commonly used fluorescent dye rhodamine B (RB) and PVA are selected as the acceptor and the polymeric matrix, respectively. Remarkably, the co‐doped films exhibit ultra‐long green circular polarization phosphorescence lasting several seconds with high efficiency (14.23%), lifetime (1025.5 ms), and dissymmetry factor (8.44 × 10−3). Crucially, by adjusting the acceptor content, it can precisely tune the phosphorescence‐to‐fluorescence ratio and achieve multicolor afterglow. This control stems from a combination of Förster resonance energy transfer, chirality transfer, and color‐mixing effects. Furthermore, this approach significantly improves the asymmetry factor of the CPL afterglow by an order of magnitude (5.17 × 10−2). It is anticipated that this straightforward approach to precisely regulating the afterglow color of CPL will provide invaluable insights for the future design of highly efficient and innovative CPL materials. [ABSTRACT FROM AUTHOR]

Styrylpyridinium dye bearing azadithia‐15‐crown‐5 ether receptor group SP and 4‐alkoxy‐1,8‐naphthalimide fluorophore were linked using copper‐catalyzed azide‐alkyne cycloaddition click reaction to afford dyad compound NI‐SP. Chemosensor NI‐SP exhibited selective ratiometric fluorescent response to the presence of Hg2+ in aqueous solution due to the interplay between resonance energy transfer (RET) and intramolecular charge transfer (ICT) processes occurred upon excitation. The observed switching of the ratio of emission intensities in the blue and red channels R was higher than in the case of monochromophoric styrylpyridine derivative SP showing ratiometric response based on ICT mechanism only. Biological studies revealed that NI‐SP penetrates into human lung adenocarcinoma A549 cells and accumulates in cytoplasm and lysosomes. When cells were pre‐incubated with mercury (II) perchlorate, the ratio R was increased 2.6 times, which enables detection of intracellular Hg2+ ions and their quantitative analysis in the 0.7–6.0 μM concentration range. [ABSTRACT FROM AUTHOR]

The development of rewritable information encryption technology with high security is essential to meet the increasing confidentiality needs, however, it remains challenging. Hence, a strategy using self‐growth technology and host–guest interaction to construct reprogrammable photoswitchable supramolecular fluorescent polymers (PSFP) is developed, enabling rewritable information encryption. In this strategy, the polymerizable adamantane (AD) is integrated into polymer materials by employing the self‐growth method to create the desired 3D structure on the surface of the matrix. The fluorescent dyes modified by β‐cyclodextrin (CDPA and CDNA) and photochromic spiropyran‐linked β‐cyclodextrin (CDSP) are incorporated into the growing structure through host–guest interactions between AD unit and β‐cyclodextrin (β‐CD) moieties. Based on the photo‐induced fluorescence resonance energy transfer (FRET) process, PSFP can undergo reversible dual‐state fluorescence switching between blue/green and red. Interestingly, owing to the dynamic features of host–guest interactions, the fluorescent information can be erased and rewritten easily. These outstanding performances promote to apply PSFP to rewritable information encryption and anti‐counterfeiting labels. Overall, this strategy opens up new prospects for smart photoswitchable fluorescent materials. [ABSTRACT FROM AUTHOR]

Background and Purpose Experimental Approach Key Results Conclusion and Implications Orexins have important biological effects on the central and peripheral nervous systems. Their primary ability is to regulate the sleep–wake cycle. Orexins and their antagonists, via OX1 receptor have been shown to have proapoptotic and antitumor effects on various digestive cancers cell models. We investigated, (1) the ability of orexin‐A and its antagonists to regulate OX1 receptor expression at the cell surface and (2), how OX1 antagonists induced proapoptotic effect in cancer cells models.The OX1 receptor internalisation is determined by imaging flow cytometry in colon cancer cell models and the OX1 receptor coupling to G proteins via bioluminescence resonance energy transfer and molecular dynamic simulation.Orexin‐A induced rapid receptor internalisation within 15 min via β‐arrestin 2 recruitment, whereas antagonists had no effect. Furthermore, Gq is critical for receptor internalisation and signalling pathways, and no other G proteins appear to be recruited. Surprisingly, antagonists induced recruitment and conformational changes in Gq protein. Simulated molecular dynamics of agonists/orexin receptor/Gq complexes show that antagonists exhibits a similar binding mode, stable at the binding site and show conformational changes of ECL2, similar to that of the agonists.OX1 receptor activation induced orexin/β‐arrestin‐dependent internalisation, which was independent of the apoptotic pathway induced by orexins and antagonists. In addition, antagonists activate the Gq protein, suggesting its putative partial dissociation. These results suggest that the development of OX1 receptor targeting molecules, including orexin antagonists with antitumor properties, may pave the way for innovative cancer therapies. [ABSTRACT FROM AUTHOR]

[Display omitted] Förster resonance energy transfer (FRET) serves as a critical mechanism to study intermolecular interactions and the formation of macromolecular assemblies. Cascade FRET is a multi-step FRET process which can overcome limitations associated with traditional single-step FRET. Herein, a novel organic–inorganic hybrid composed of a nile red derivative attached to the edge of the layered silicate clay Laponite (Lap-NR) was used to facilitate cascade FRET between Laponite sheets. Utilizing naphthalene-diimide edge-modified Laponite (Lap-NDI) as the initial donor, Rhodamine 6G on the basal surface of Laponite as the first acceptor, and Lap-NR as the second acceptor, cascade FRET was achieved. The influence of solvent composition in a DMF/water mixture on cascade FRET was investigated, revealing that a higher water content led to an enhancement of the cascade FRET process, which is attributed to increased aggregation-induced emission of Lap-NDI and the enhanced quantum yield of R6G in water. This study provides a unique approach to achieve cascade FRET by taking advantage of the anisotropic surface chemistry of a two-dimensional nanomaterial, providing a colloidally-driven alternative with improved tunability compared to macromolecular routes. The flexibility and simplicity of this approach will advance the state of the art of organic–inorganic hybrids for applications in optoelectronics, sensors, and hybrid photovoltaics. [ABSTRACT FROM AUTHOR]

Intrinsically disordered proteins (IDPs) adopt ensembles of rapidly fluctuating heterogeneous conformations, influencing their binding capabilities and supramolecular transitions. The primary conformational descriptors for understanding IDP ensembles--the radius of gyration (RG), measured by small-angle X-ray scattering (SAXS), and the root mean square (rms) end-to-end distance (RE), probed by fluorescent resonance energy transfer (FRET)--are often reported to produce inconsistent results regarding IDP expansion as a function of denaturant concentration in the buffer. This ongoing debate surrounding the FRET-SAXS discrepancy raises questions about the overall reliability of either method for quantitatively studying IDP properties. To address this discrepancy, we introduce a genetically encoded anomalous SAXS (ASAXS) ruler, enabling simultaneous and direct measurements of RG and RE without assuming a specific structural model. This ruler utilizes a genetically encoded noncanonical amino acid with two bromine atoms, providing an anomalous X-ray scattering signal for precise distance measurements. Through this approach, we experimentally demonstrate that the ratio between RE and RG varies under different denaturing conditions, highlighting the intrinsic properties of IDPs as the primary source of the observed SAXS-FRET discrepancy rather than shortcomings in either of the two established methods. The developed genetically encoded ASAXS ruler emerges as a versatile tool for both IDPs and folded proteins, providing a unified approach for obtaining complementary and site-specific conformational information in scattering experiments, thereby contributing to a deeper understanding of protein functions. [ABSTRACT FROM AUTHOR]

Autophagy is a ubiquitous process of organelle interaction in eukaryotic cells, in which various organelles or proteins are recycled and operated through the autophagy pathway to ensure nutrient and energy homeostasis. Although numerous fluorescent probes have been developed to image autophagy, these environment‐responsive probes suffer from inherent deficiencies such as inaccuracy and limited versatility. Here, we present a modular macrocyclic amphiphile Förster Resonance Energy Transfer (FRET) platform (SC6A12C/NCM, SN), constructed through the amphiphilic assembly of sulfonatocalix[6]arene (SC6A12C) with N‐cetylmorpholine (NCM) for lysosome targeting. The hydrophobic fluorophore BPEA (FRET donor) was entrapped within the inner hydrophobic phase and showed strong fluorescence emission. Attributed to the broad‐spectrum encapsulation of SC6A12C, three commercially available organelle probes (Mito‐Tracker Red, ER Tracker Red, and RhoNox‐1) were selected as SC6A12C guests (FRET acceptors). During autophagy process, the formation of intracellular host–guest complexes leads to strong FRET signal, allowing us to visualize the fusion of mitochondria, endoplasmic reticulum, and Golgi apparatus with lysosomes, respectively. This study provides a versatile and accessible platform for imaging organelle autophagy. [ABSTRACT FROM AUTHOR]

Cyclic nucleotide-binding domain (CNBD) ion channels play crucial roles in cellular-signaling and excitability and are regulated by the direct binding of cyclic adenosine- or guanosine-monophosphate (cAMP, cGMP). However, the precise allosteric mechanism governing channel activation upon ligand binding, particularly the energetic changes within domains, remains poorly understood. The prokaryotic CNBD channel SthK offers a valuable model for investigating this allosteric mechanism. In this study, we investigated the conformational dynamics and energetics of the SthK C-terminal region using a combination of steady-state and time-resolved transition metal ion Förster resonance energy transfer (tmFRET) experiments. We engineered donor-acceptor pairs at specific sites within a SthK C-terminal fragment by incorporating a fluorescent noncanonical amino acid donor and metal ion acceptors. Measuring tmFRET with fluorescence lifetimes, we determined intramolecular distance distributions in the absence and presence of cAMP or cGMP. The probability distributions between conformational states without and with ligand were used to calculate the changes in free energy (ΔG) and differences in free energy change (ΔΔG) in the context of a simple four-state model. Our findings reveal that cAMP binding produces large structural changes, with a very favorable ΔΔG. In contrast to cAMP, cGMP behaved as a partial agonist and only weakly promoted the active state. Furthermore, we assessed the impact of protein oligomerization and ionic strength on the structure and energetics of the conformational states. This study demonstrates the effectiveness of time-resolved tmFRET in determining the conformational states and the ligand-dependent energetics of the SthK C-terminal region. [ABSTRACT FROM AUTHOR]

Stimulus‐responsive fluorescent polymeric materials (FPMs) that can change their fluorescent states via external stimuli have been widely applied in multi‐level information encryption and anti‐counterfeiting. While many efforts have mainly focused on the design of dual‐stimuli responsive FPMs, the construction of orthogonal photo‐ and thermo‐regulation of fluorescent polymeric systems remains a challenge. Here, the orthogonal photo‐ and thermo‐responsive fluorescent polymeric hydrogels (PTFPHs) containing capsules with the phase change materials (PCMs), photoresponsive molecules (DTE), and thermally responsive molecules (TPA‐DCPP) are reported. They are capable of reconfiguring fluorescence (none, green, yellow, red) via photo‐ and thermo‐induced fluorescence resonance energy transfer (FRET) process. Additionally, not only can the thermochromic property of PTFPHs be regulated by modulating the composition of PCMs, but the thermochromic and photochromic properties of PTFPHs can also be tuned by using SDS to control the size of capsules. The developed hydrogels exhibited high fluorescence contrast, fast response, and excellent reversibility. The PTFPHs are successfully applied in multi‐level information encryption and integrated application between temperature monitoring and anti‐counterfeiting. The works represented a strategy for developing multi‐stimuli responsive PTFPHs in multi‐level information encryption and advanced anti‐counterfeiting. [ABSTRACT FROM AUTHOR]