Alternative Mechanisms Research Articles

Robust lipid scrambling by TMEM16 proteins requires an open groove and thin membrane.

Biological membranes are complex and dynamic structures with different populations of lipids on their inner and outer leaflets. The Ca2+-activated TMEM16 family of membrane proteins play an important role in collapsing this asymmetric lipid distribution by spontaneously, and bidirectionally, scrambling phospholipids between the two leaflets, which can initiate signaling and alter the physical properties of the membrane. While evidence shows that lipid scrambling occurs via an open hydrophilic pathway ("groove") that spans the membrane, it remains unclear if all family members facilitate lipid movement in this manner. Here we present a comprehensive computational study of lipid scrambling by all TMEM16 members with experimentally solved structures. We performed coarse-grained molecular dynamics (MD) simulations of 27 structures from five different family members solved under activating and non-activating conditions, and we captured over 700 scrambling events in aggregate. This enabled us to directly compare scrambling rates, mechanisms, and protein-lipid interactions for fungal and mammalian TMEM16s, in both open (Ca2+-bound) and closed (Ca2+-free) conformations with statistical rigor. We show that nearly all (>90%) scrambling occurs in the dilated groove and the degree of induced membrane thinning positively correlates with the scrambling rate. Surprisingly, we also observed 60 scrambling events that occurred outside the canonical groove, over 90% of which took place at the dimer-dimer interface in mammalian TMEM16s. This new site suggests an alternative mechanism for lipid scrambling in the absence of an open groove.

Triagem in silico de compostos com atividade ansiolítica encontrados na espécie Magnolia obovata

Magnolia obovata, known as “Japanese cucumber”, is a deciduous tree of Asian origin, constituting a medicinal plant due to its anti-inflammatory, anxiolytic, antidepressant effects, among other central effects, already demonstrated in the literature. The objective of this study was to suggest the mechanisms of action for the effects on the central nervous system of the compounds identified in the species M. obovata, especially regarding the anxiolytic effect currently sought with the use of the plant. Nineteen compounds present in M. obovata were identified, with only 2 molecules (alpha-eudesmol and gamma-eudesmol) showing in silico pharmacokinetic and toxicological properties favorable to anxiolytic bioactivity. Such molecules inhibit acylcarnitine hydrolase and increase free acylcarnitine, possibly generating an anxiolytic effect. Pharmacophoric modeling of those molecules showed 6 interaction points with the 5 most potent known ligands of acylcarnitine hydrolase and such structural similarity is promising for acting on this target. There are advantages of the alternative mechanism of action of this compound in relation to current anxiolytics, which could be used to formulate new therapies in the treatment of anxiety disorders. The results obtained here open perspectives for tests in in vitro and in vivo models, aiming to confirm the results of the computational analyses.

The impact of freeze-dried Baiyedancong-Oolong tea aqueous extract containing bioactive compounds on the activities of CYP450 enzymes, the transport capabilities of P-gp and OATs, and transcription levels in mice.

In this study, (-)-epigallocatechin gallate (EGCG) and caffeine extracted from freeze-dried autumn Baiyedancong Oolong tea (FBOT) were orally administered to mice for 7 consecutive days to explore the effects of BOT and its bioactive compounds on the activities and transcription levels of CYP450 enzymes, intestinal effluence transporter P-gp, and renal ingestion Organic Anion Transporters (OATs). Concurrently, EGCG and caffeine enhanced the activities of CYP3A, CYP2E1, and CYP2C37 in the liver of mice, while impairing the transport capabilities of P-gp and OATs. Reduced levels of MDR1 encoding P-gp transcription in the small intestine and renal OAT1 and OAT3 revealed that transcription was involved in the regulation of CYP450, P-gp, and OATs. The reduced transcription level of liver CYP2E1 suggested that CYP2E1 activity may have been elevated due to alternative mechanisms, but not through transcription. The absorption, metabolism, and excretion of drugs may be influenced by the daily consumption or high-dose administration of BOT and its related products, in which EGCG and caffeine may make great contributions.

C-4 analogues of podophyllotoxin as tubulin inhibitors: synthesis, biological evaluation, and structure-activity relationship

The diversity of lignan small molecules derived from podophyllotoxin, a non-covalent tubulin inhibitor isolated from the Podophyllum family, has led to the clinical development of FDA-approved anticancer agents etoposide and teniposide. While these two compounds share the same tetracyclic core as podophyllotoxin, two subtle structural changes—4’ demethylation on the aromatic ring and stereospecific glycosylation at the C-4 hydroxyl—result in an alternate biological mechanism. Given the immense pharmacological importance of altering the C-4 position, we synthesised and evaluated a systematic library of diversified esters to establish a structure-activity relationship regarding modification at C-4 on the properties of podophyllotoxin. We determined the biological activity of these esters through cell viability assays, computer docking models, tubulin polymerisation assays, and cell cycle analysis. Altogether, we demonstrate that increasing steric hindrance at C-4 leads to a loss in potency against human cancer cells but has a significantly lesser impact on cell-free tubulin inhibition.

Expression of Manganese Transporters ZIP8, ZIP14, and ZnT10 in Brain Barrier Tissues.

Manganese (Mn) is an essential trace mineral for brain function, but excessive accumulation can cause irreversible nervous system damage, highlighting the need for proper Mn balance. ZIP14, ZnT10, and ZIP8 are key transporters involved in maintaining Mn homeostasis, particularly in the absorption and excretion of Mn in the intestine and liver. However, their roles in the brain are less understood. The blood-cerebrospinal fluid barrier and the blood-brain barrier, formed by the choroid plexus and brain blood vessels, respectively, are critical for brain protection and brain metal homeostasis. This study identified ZIP14 on the choroid plexus epithelium, and ZIP8 and ZnT10 in brain microvascular tissue. We show that despite significant Mn accumulation in the CSF of Znt10 knockout mice, ZIP14 expression levels in the blood-cerebrospinal fluid barrier remain unchanged, indicating that ZIP14 does not have a compensatory mechanism for regulating Mn uptake in the brain in vivo. Additionally, Mn still enters the CSF without ZIP14 when systemic levels rise. This indicates that alternative transport mechanisms or compensatory pathways ensure Mn balance in the CSF, shedding light on potential strategies for managing Mn-related disorders.

RNAseq-Based Carboxylesterase Nl-EST1 Gene Expression Plasticity Identification and Its Potential Involvement in Fenobucarb Resistance in the Brown Planthopper Nilaparvata lugens.

Carbamate insecticides have been used for over four decades to control brown planthopper, Nilaparvata lugens, but resistance has been reported in many countries, including the Republic of Korea. The bioassay results on resistance to fenobucarb showed that the LC50 values were 3.08 for the susceptible strain, 10.06 for the 2015 strain, and 73.98 mg/L for the 2019 strain. Compared to the susceptible strain, the 2015 and 2019 strains exhibited resistance levels 3.27 and 24.02 times higher, respectively. To elucidate the reason for the varying levels of resistance to fenobucarb in these strains, mutations in the acetylcholinesterase 1 (ACE1) gene, the target gene of carbamate, were investigated, but no previously reported mutations were confirmed. Through RNA-seq analysis focusing on the expression of detoxification enzyme genes as an alternative resistance mechanism, it was found that the carboxylesterase gene Nl-EST1 was overexpressed 2.4 times in the 2015 strain and 4.7 times in the 2019 strain compared to the susceptible strain. This indicates a strong correlation between the level of resistance development in each strain and the expression level of Nl-EST1. Previously, Nl-EST1 was reported in an organophosphorus insecticide-resistant strain of Sri Lanka 2000. Thus, Nl-EST1 is crucial for developing resistance to organophosphorus and carbamate insecticides. Resistance-related genes such as Nl-EST1 could serve as expression markers for resistance diagnosis, and can apply to integrated resistance management of N. lugens.

Rapid Response Mode observations of GRB 160203A: Looking for fine-structure line variability at z = 3.52

Context. Gamma-ray bursts are the most energetic known explosions. Although they fade rapidly, they give us the opportunity to measure redshift and important properties of their host galaxies. We report the photometric and spectroscopic study of the Swift GRB 160203A at z = 3.518, and its host galaxy. Fine-structure absorption lines, detected in the afterglow at different epochs, allow us to investigate variability due to the strong fading background source. Aims. We obtained two optical to near-infrared spectra of the GRB afterglow with X-shooter on ESO/VLT, 18 minutes and 5.7 hours after the burst, allowing us to investigate temporal changes of fine-structure absorption lines. Methods. We measured H I column density log N(HI/cm–2) = 21.75 ± 0.10, and several heavy-element ions along the GRB sightline in the host galaxy, among which Si II, Al II, Al III, C II, Ni II, Si IV, C IV, Zn II and Fe II, and Fe II∗ and Si II∗ fine-structure transitions from energetic levels excited by the afterglow, at the common redshift z = 3.518. We measured [M/H]TOT = –0.78 ± 0.13 and a [Zn/Fe]FIT = 0.69 ± 0.15, representing the total (dust corrected) metallicity and dust depletion, respectively. We detected additional intervening systems along the line of sight at ɀ = 1.03, ɀ = 1.26, ɀ = 1.98, ɀ = 1.99, ɀ = 2.20, and ɀ = 2.83. We could not measure significant variability in the strength of the fine-structure lines throughout all the observations and determined an upper limit for the GRB distance from the absorber of d < 300 pc, adopting the canonical UV pumping scenario. However, we note that the quality of our data is not sufficient to conclusively rule out collisions as an alternative mechanism. Results. GRB 160203A belongs to a growing sample of GRBs with medium resolution spectroscopy, provided by the Swift/X-shooter legacy programme, which enables a detailed investigation of the interstellar medium in high-redshift GRB host galaxies. In particular, this host galaxy shows relatively high metal enrichment and dust depletion already in place when the universe was only 1.8 Gyr old.

Revisiting oxygen toxicity: evolution and adaptation to superoxide in a SOD-deficient bacterial pathogen.

Defenses against oxidants are crucial for the virulence of pathogens, with superoxide scavenging enzymes (SOSEs) playing a vital role for most aerobes. However, our knowledge of superoxide adaptation primarily stems from the study of SOSE-encoding bacteria. Here, we investigated the evolution of a naturally SOSE-deficient pathogen ( Leptospira spp.), along with the alternative mechanisms it recruits to combat superoxide stress. We demonstrate that emergence of pathogenic Leptospira correlated with SOD loss, but that a long-lasting adaptation to superoxide remains possible. We reveal that cysteine and leucine biosynthesis are the most induced pathways in response to superoxide and demonstrate the importance of sulfur metabolism in superoxide adaptation in this SOSE-deficient model. We also propose cysteine oxidation as a key mediator of superoxide toxicity in the absence of SOSEs. This study challenges our conventional understanding of the oxygen toxicity theory and proposes a new model of superoxide adaptation through metabolic rewiring in bacteria.

The role of pigments in light color variation of the firefly Photinus pyralis.

Fireflies use bioluminescent signals to communicate with their mates. Luciferase has been thought to be the sole contributor to light color; however, populations of the Photinus pyralis firefly display variation in the color of their emitted signals yet have identical luciferase sequences. Here, we examined whether pigments could be present in the light organs of the twilight-active species P. pyralis and contribute to this variation. We detected patterns of expression that suggest ommochrome and pterin screening pigments are expressed in P. pyralis light organs and could filter light emitted by luciferase and play a role in signal tuning. There were no significant differences between the pigment gene expression of P. pyralis individuals with yellower and greener signals. Our study provides alternative mechanisms that could influence pigments in P. pyralis light organs that could also play a role in modifying signal color.

The deformation mechanism of low symmetric Ti–Pt intermetallic compounds containing high density of planar defects

Intermetallic compounds typically exhibit limited plastic deformation capacity due to challenges in activating dislocation slip and deformation twinning, coupled with a lack of alternative deformation mechanisms. Ti–Pt alloys are a prevalent type of intermetallic compound utilized in high-temperature shape memory alloys and as materials for energy applications in electric fields. However, they often exhibit poor deformation capability. Here, we prepared a low-symmetry intermetallic phase, Ti4Pt3, which demonstrates significant plastic deformation capability. This phase features a high density of parallel planar defects, resulting in an exceptionally large lattice periodicity perpendicular to these defects. Through in-situ scanning electron microscope compression tests, we observed substantial plastic deformation in this new phase. Analysis of the deformed Ti4Pt3 phase revealed that the dense planar defects create uniformly distributed sites of internal stress concentration, enabling a rapid increase in back stress within crystals. This phenomenon leads to notable lattice rotation and localized order-disorder transitions, both crucial mechanisms facilitating plastic deformation and enhancing deformation capacity. Our research underscores the potential of leveraging structural asymmetry to enable unconventional deformation mechanisms, thereby enhancing the plasticity of intermetallic materials.

PITAR, a DNA damage-inducible cancer/testis long noncoding RNA, inactivates p53 by binding and stabilizing TRIM28 mRNA.

In tumors with WT p53, alternate mechanisms of p53 inactivation are reported. Here, we have identified a long noncoding RNA, PITAR (p53 Inactivating TRIM28 Associated RNA), as an inhibitor of p53. PITAR is an oncogenic Cancer/testis lncRNA and is highly expressed in glioblastoma (GBM) and glioma stem-like cells (GSC). We establish that TRIM28 mRNA, which encodes a p53-specific E3 ubiquitin ligase, is a direct target of PITAR. PITAR interaction with TRIM28 RNA stabilized TRIM28 mRNA, which resulted in increased TRIM28 protein levels and reduced p53 steady-state levels due to enhanced p53 ubiquitination. DNA damage activated PITAR, in addition to p53, in a p53-independent manner, thus creating an incoherent feedforward loop to inhibit the DNA damage response by p53. While PITAR silencing inhibited the growth of WT p53 containing GSCs in vitro and reduced glioma tumor growth in vivo, its overexpression enhanced the tumor growth in a TRIM28-dependent manner and promoted resistance to Temozolomide. Thus, we establish an alternate way of p53 inactivation by PITAR, which maintains low p53 levels in normal cells and attenuates the DNA damage response by p53. Finally, we propose PITAR as a potential GBM therapeutic target.

The Russian–Ukrainian war: The possibilities of ensuring accountability

AbstractThe outbreak of the Russian–Ukrainian war fundamentally shook the international legal order. Consequently, possibilities of holding the aggressor state and potential perpetrators of international crimes accountable became a central topic. Options for establishing state responsibility for violating international law are limited. Possible individual criminal prosecution mechanisms are available at both national and international levels. At the national level, several jurisdictional bases can be found for prosecuting the most serious international crimes.In addition to the procedures at the national level, the International Criminal Court has jurisdiction at the international level in terms of holding persons accountable; however, this also faces challenges. In connection with the current conflict, the International Criminal Court cannot conduct proceedings in relation to all categories of crimes, so the international community seeks to rely on alternative mechanisms that support prosecution efforts. Furthermore, there is an increasing political demand for the creation of new mechanisms which, however, for the time being, are based on shaky international legal grounds.

Перспективы рынка киберстрахования США под влиянием новых вызовов

The article delves into the analysis of current trends in the cyber risk insurance market in the United States, which is both the driving force of the global insurance industry and a major victim of cyber attacks. The work highlights the key factors that define the current cyber insurance market landscape and influence the increasing frequency and severity of cyber attacks. Legislative initiatives to curb the activities of attackers and create an effective infrastructure to combat ransomware developers are examined, which in turn determine insurers' underwriting policies when assessing cyber risks. The research emphasizes the growth of alternative risk transfer mechanisms and instruments, as well as the increasing restrictions on underwriting risks of a systemic nature against the backdrop of escalating cyber threats. The article presents an assessment of the current state and outlines the prospects for the development of the US cyber insurance market with a focus on improving modeling mechanisms and attracting highly specialized personnel.

Population differences in putty-nosed monkey (Cercopithecus nictitans) call order.

Non-human primates generally lack the ability to learn new call structures or to substantially modify existing ones, suggesting that callers need alternative mechanisms to convey information. One way to escape the constraints of limited vocal control is by assembling calls into variable sequences, as has been documented in various animal species. Here, we were interested in the flexibility with which different calls might be assembled in a species known for its meaningful call order, putty-nosed monkeys (Cercopithecus nictitans). Since most information comes from studies conducted at Gashaka Gumti National Park (Nigeria), we tested two further populations in the Nouabalé-Ndoki National Park (Republic of the Congo) and Taï National Park (Côte d'Ivoire) in how males responded to common threats, leopards, and crowned eagles. As predicted, callers produced the same basic call types as seen elsewhere-long 'pyow', short 'pyow' ('kek'), 'hack'-but populations differed in how males assembled calls. To leopards, males from both populations started with 'pyows' and 'keks', with occasional hacks later, as already reported from Gashaka. To crowned eagle, however, Nouabalé-Ndoki males consistently initiated their responses with 'pyows', whereas neither Taï nor Gashaka males ever did, demonstrating that nonhuman primates have some control over sequence production. We discuss possible mechanisms to account for the population differences, predation pressure, and male-male competition, and address implications for linguistic theories of animal call order, notably the Urgency and Informativity Principles.

Salt-Induced Hydrophobic C-Terminal Region of α-Synuclein Triggers Its Fibrillation under the Mimic Physiologic Condition.

α-Synuclein (αS) causes Parkinson's disease due to the structural alteration into amyloid fibrils that form after the interaction with synaptic membranes in neurons. To understand the alternation mechanism, the effect of salt (NaCl) on the interaction of αS with synaptic mimic membrane was characterized at the molecular level because salt triggered the amyloid fibril formation. The membrane-bound conformation (or the initial conformation before fibrillation) showed that NaCl decreased the number of helical structures and Tyr residues interacting with the membrane surface compared to when NaCl was absent, implying an increase in solvent-exposed regions. The N-terminal region of αS interacted with the membrane, forming the helical structures regardless of NaCl, while the C-terminal region formed a random structure with weak membrane interaction, but NaCl inhibited the interaction of its hydrophobic area, suggesting that salt promoted amyloid fibril formations by exposing the hydrophobic C-terminal region, which can intermolecularly interact with free αS.

Coherent spin wave excitation with radio-frequency spin–orbit torque

Spin waves, collective perturbations of magnetic moments, are both fundamental probes for magnetic physics and promising candidates for energy-efficient signal processing and computation. Traditionally, coherent propagating spin waves have been generated by radio frequency (RF) inductive Oersted fields from current-carrying electrodes. An alternative mechanism, spin–orbit torque (SOT), offers more localized excitation through interfacial spin accumulation but has been mostly limited to DC to kHz frequencies. SOT driven by RF currents, with potentially enhanced pumping efficiency and unique spin dynamics, remains largely unexplored, especially in magnetic insulators. Here, we conduct a comprehensive theoretical and computational investigation into the generation of coherent spin waves via RF-SOT in the prototypical yttrium iron garnet. We characterize the excitation of forward volume, backward volume, and surface modes in both linear and nonlinear regimes, employing single and interdigitated electrode configurations. We reveal and explain several unique and surprising features of RF-SOT compared to inductive excitation, including higher efficiency, distinct mode selectivity, and directional symmetry, a ∼3π/4 phase offset, reduced anharmonic distortion in the nonlinear regime, and the absence of second harmonic generation. These insights position RF-SOT as a promising new mechanism for future magnonic and spintronic applications.

Insomnia symptoms as long-term predictors of anxiety symptoms in middle-aged and older adults from the English Longitudinal Study of Ageing (ELSA), and the role of systemic inflammation

Insomnia, i.e., difficulties in sleep onset and sleep maintenance, may increase the risk of anxiety symptoms, although long-term follow-up studies are rarely reported. Here, we examined whether insomnia symptoms may predict anxiety symptoms in a 9-year follow-up, and whether inflammation may play a mediating role. Data from 1355 participants (63.44 ± 7.47 years, 55.1 % females) from the English Longitudinal Study of Ageing (ELSA) were analysed. Insomnia symptoms were assessed in 2012/13. High-sensitivity C-reactive protein (hs-CRP), a marker of systemic inflammation, was measured in 2016/17. Anxiety symptoms were assessed in 2020/21. After adjusting for confounders and baseline levels, structural equation modelling (SEM) revealed that insomnia symptoms significantly predicted anxiety symptoms (β = 0.357, p < .001) but not hs-CRP (β = −0.016, p = .634). Similarly, hs-CRP was not related to anxiety symptoms (β = −0.024, p = .453). The hs-CRP mediation hypothesis was therefore rejected (β = 0.0004; 95 % BCI -0.001 to 0.005), and multi-group SEM showed that sex did not moderate these paths. However, baseline diagnoses of anxiety disorders prospectively predicted higher hs-CRP (B = 0.083, p = .030). Results of the current study suggest that individuals with baseline anxiety disorders may be at higher risk of developing low-grade chronic inflammation. Several alternative psychophysiological mechanisms linking insomnia and anxiety symptoms should be explored, including autonomic and cortical pre-sleep arousal, cortisol reactivity, and pro-inflammatory cytokines. Finally, insomnia symptoms may be a treatment target to lower the risk of anxiety symptoms in elderly.

Sub-Boltzmann Switching, Hysteresis-Free Charge Modulated Negative Differential Resistance FinFET.

The ever-increasing power consumption in integrated circuits has raised concerns about the relentless doubling of transistor density in chips and cost drop per combinational/sequential circuits. To address the physical limit of thermionic emission carrier transport (i.e., subthreshold swing >60 mV/decade at 300 K), alternative charge-transport mechanisms or the implementation of functional substances have been attempted but without appreciable success. One such choice is to take advantage of negative differential resistance with the activation of localized electrons or migration of atom and oxygen vacancies to extend the capabilities of Si-transistors. However, inconsistency in current during forward/reverse bias sweep is confronted as a notable weak point. This work proposes an eye-catching solution to modulating potential distribution between a resistance switching layer and a transistor by employing charge trapping within a hafnium zirconium oxide layer. This approach introduces features advancing the potential of "More Moore" technologies.

Plasma glucosylceramide levels are regulated by ATP10D and are not involved in Parkinson's disease pathogenesis.

GBA1 variants and decreased glucocerebrosidase (GCase) activity are implicated in Parkinson's disease (PD). We investigated the hypothesis that increased levels of glucosylceramide (GlcCer), one of GCase main substrates, are involved in PD pathogenesis. Using multiple genetic methods, we show that ATP10D, not GBA1, is the main regulator of plasma GlcCer levels, yet it is not involved in PD pathogenesis. Plasma GlcCer levels were associated with PD, but not in a causative manner, and are not predictive of disease status. These results argue against targeting GlcCer in GBA1-PD and underscore the need to explore alternative mechanisms and biomarkers for PD.

No such thing as waste in primary food sector.

The European Union (EU) circular economy action plan aims to double its use of recycled material by 2030. We argue that waste-centric approach to resources may have adverse consequences to this ambition. The aim of the work was to find out the factors limiting or promoting the use of waste from primary food sector in countries with the same cultural background and similar climate in Estonia, Latvia, Lithuania and Norway. Biomass from the primary food production sector is of good quality and excellent to use, but its use may be limited if given waste status. From numerous management decision trigger clusters, which may affect valorisation of the biomass, we focus on technology and policy. Our semi-structured interviews addressed the analysis and management of waste or by-products and explored the end-of-waste and alternative mechanisms that allowed the biomass to be valorised. However, the interviews revealed that none of the companies regard anything becoming waste, but as raw material or production left-over. Any obstacles hypothesised turned out not to be acknowledged by companies at all. This appears to be a very good example of the use of resources, but the approach is haphazard and may conflict with official understanding and waste reporting requirements. Definition of waste is the same in the EU, and arbitrary treatment can be misleading. There is a need for better management of the material flow to ensure effective biomass circulation avoiding its becoming waste. We recommend that this be addressed by introducing environmental, social, governance and a self-control system.