Mechanistic Insights Research Articles

Modulating Polymerase Activity through Light-Oxygen-Voltage Domain Insertion.

Biochemical reaction networks adapt to environmental conditions by sensing chemical or physical stimuli and using tightly controlled mechanisms. While most signals come from molecules, many cells can also sense and respond to light. Among the biomolecular structures that enable light sensing, we selected a light-oxygen-voltage (LOV) domain in a previous study that tested the engineering of novel regulatory mechanisms into a nucleic acid polymerase. In this follow-up study, we studied the activities of previously selected variants in kinetic detail, and we generated additional LOV-polymerase fusion variants based on further insertion criteria. Our results provide mechanistic insights into how LOV domain insertion influences polymerase activity in a light-responsive manner: All active and photoresponsive enzyme variants studied by us to date were partially inhibited (i.e., "turned off") after irradiation with blue light at 470 nm, which can be explained by specific obstructions of the polymerase entry or exit structures (substrate entry channels or product exit channels, or both). Although the effects observed are moderate, we anticipate further engineering strategies that could be used to improve the extent of switchability and possibly to develop a "turn-on mode" insertion.

Frontostriatal salience network expansion in individuals in depression

Decades of neuroimaging studies have shown modest differences in brain structure and connectivity in depression, hindering mechanistic insights or the identification of risk factors for disease onset1. Furthermore, whereas depression is episodic, few longitudinal neuroimaging studies exist, limiting understanding of mechanisms that drive mood-state transitions. The emerging field of precision functional mapping has used densely sampled longitudinal neuroimaging data to show behaviourally meaningful differences in brain network topography and connectivity between and in healthy individuals2–4, but this approach has not been applied in depression. Here, using precision functional mapping and several samples of deeply sampled individuals, we found that the frontostriatal salience network is expanded nearly twofold in the cortex of most individuals with depression. This effect was replicable in several samples and caused primarily by network border shifts, with three distinct modes of encroachment occurring in different individuals. Salience network expansion was stable over time, unaffected by mood state and detectable in children before the onset of depression later in adolescence. Longitudinal analyses of individuals scanned up to 62 times over 1.5 years identified connectivity changes in frontostriatal circuits that tracked fluctuations in specific symptoms and predicted future anhedonia symptoms. Together, these findings identify a trait-like brain network topology that may confer risk for depression and mood-state-dependent connectivity changes in frontostriatal circuits that predict the emergence and remission of depressive symptoms over time.

Silver-π Interaction: A Diverse Approach to Hybrid Material and Its Efficacy in Electrocatalytic Reduction of Nitrate to Ammonia.

Inspired by the intriguing nature of the metal-π interaction in organometallic chemistry, a novel 1D hybrid material has been designed. Herein, a functionalized tellurium allyl macrocycle (TAM) acts as a molecular building block and is knit together via silver-π interaction to obtain Ag-TAM. Ag is coordinated to two allyl groups and a phenyl ring in η2 mode. Instead of the conventional polymerization strategy, a metal-π interaction is employed to interlink macrocycles. TAM and Ag-TAM showed electrocatalytic capability for the conversion of nitrate to ammonia. Ag-TAM showed an NH3 yield rate 2-fold greater than TAM with a high faradaic efficiency of 94.6% with good durability, proving that interlinking of macrocycles via metal-π interaction improves the catalytic activity. Detailed periodic density functional theory (DFT) calculations unveil novel mechanistic insights, suggesting cooperative catalysis between neighboring Ag sites and contributing to the enhanced efficiency.

Efficient Energy and Electron Transfer Photocatalysis with a Coulombic Dyad.

Photocatalysis holds great promise for changing the way value-added molecules are currently prepared. However, many photocatalytic reactions suffer from quantum yields well below 10%, hampering the transition from lab-scale reactions to large-scale or even industrial applications. Molecular dyads can be designed such that the beneficial properties of inorganic and organic chromophores are combined, resulting in milder reaction conditions and improved reaction quantum yields of photocatalytic reactions. We have developed a novel approach for obtaining the advantages of molecular dyads without the time- and resource-consuming synthesis of these tailored photocatalysts. Simply by mixing a cationic ruthenium complex with an anionic pyrene derivative in water a salt bichromophore is produced owing to electrostatic interactions. The long-lived organic triplet state is obtained by static and quantitative energy transfer from the preorganized ruthenium complex. We exploited this so-called Coulombic dyad for energy transfer catalysis with similar reactivity and even higher photostability compared to a molecular dyad and reference photosensitizers in several photooxygenations. In addition, it was shown that this system can also be used to maximize the quantum yield of photoredox reactions. This is due to an intrinsically higher cage escape quantum yield after photoinduced electron transfer for purely organic compounds compared to heavy atom-containing molecules. The combination of laboratory-scale as well as mechanistic irradiation experiments with detailed spectroscopic investigations provided deep mechanistic insights into this easy-to-use photocatalyst class.

Chiral Brønsted Acid-Catalyzed Intramolecular Asymmetric Dearomatization Reaction of Indoles with Cyclobutanones via Cascade Friedel-Crafts/Semipinacol Rearrangement.

The highly efficient synthesis of chiral indolines fused with an azabicyclo[2.2.1]heptanone moiety is achieved by an asymmetric dearomatization reaction of indoles with cyclobutanones. A new chiral imidodiphosphorimidate (IDPi) catalyst is synthesized and exhibits extraordinary activity in promoting a cascade Friedel-Crafts/semipinacol rearrangement. Target molecules are prepared in good yields (up to 95%) with excellent enantioselectivity (up to 98% ee) with operational convenience. Combined experimental and computational studies provide detailed mechanistic insights into the energy landscape and origin of the stereochemical induction of the reaction.

A Review on Fish Skin-Derived Gelatin: Elucidating the Gelatin Peptides-Preparation, Bioactivity, Mechanistic Insights, and Strategies for Stability Improvement.

Fish skin-derived gelatin has garnered significant attention recently due to its abundant availability and promising bioactive properties. This comprehensive review elucidates various intricacies concerning fish skin-derived gelatin peptides, including their preparation techniques, bioactive profiles, underlying mechanisms, and methods for stability enhancement. The review investigates diverse extraction methods and processing approaches for acquiring gelatin peptides from fish skin, emphasizing their impact on the peptide composition and functional characteristics. Furthermore, the review examines the manifold bioactivities demonstrated by fish skin-derived gelatin peptides, encompassing antioxidant, antimicrobial, anti-inflammatory, and anticancer properties, elucidating their potential roles in functional food products, pharmaceuticals, and nutraceuticals. Further, mechanistic insights into the functioning of gelatin peptides are explored, shedding light on their interactions with biological targets and pathways. Additionally, strategies aimed at improving the stability of gelatin peptides, such as encapsulation, modification, and integration into delivery systems, are discussed to extend the shelf life and preserve the bioactivity. Overall, this comprehensive review offers valuable insights into using fish skin-derived gelatin peptides as functional ingredients, providing perspectives for future research endeavors and industrial applications within food science, health, and biotechnology.

Radical‐Based Enantioconvergent Reductive Couplings of Racemic Allenes and Aldehydes

Transition metal‐catalyzed radical‐based enantioconvergent reactions have become a powerful strategy to synthesize enantiopure compounds from racemic starting materials. However, existing methods primarily address precursors with central chirality, neglecting those with axial chirality. Herein, we describe the enantioconvergent reductive coupling of racemic allenes with aldehydes, facilitated by a photoredox, chromium, and cobalt triple catalysis system. This method selectively affords one product from sixteen possible regio‐ and stereoisomers. The protocol leverages CoIII‐H mediated hydrogen atom transfer (MHAT) and Cr‐catalyzed radical‐polar crossover for efficient stereoablation of axial chirality and asymmetric addition, respectively. Supported by mechanistic insights from control experiments, deuterium labeling, and DFT calculations, our approach offers synthetic chemists a valuable tool for creating enantioenriched chiral homoallylic alcohols, promising to advance radical‐based strategies for synthesizing complex chiral molecules.

Corrigendum: Mechanistic Insights on the Enantioselective (4+3) cycloaddition between Oxyallylcations and Furans Catalyzed by BINOL‐based Phosphoramides

Corrigendum: Mechanistic Insights on the Enantioselective (4+3) cycloaddition between Oxyallylcations and Furans Catalyzed by BINOL‐based Phosphoramides

Multiomics analysis reveal the impact of 17α-Ethinylestradiol on mortality in juvenile zebrafish

17α-Ethinylestradiol (EE2) is known for its endocrine-disrupting effects on embryonic and adult fish. However, its impact on juvenile zebrafish has not been well established. In this study, juvenile zebrafish were exposed to EE2 at concentrations of 5 ng/L (low dose, L), 10 ng/L (medium dose, M), and 50 ng/L (high dose, H) from 21 days post-fertilization (dpf) to 49 dpf. We assessed their growth, development, behavior, transcriptome, and metabolome. The findings showed that the survival rate in the EE2-H group was 66.8 %, with all surviving fish displaying stunted growth and swollen, transparent abdomens by 49 dpf. Moreover, severe organ deformities were observed in the gills, kidneys, intestines, and heart of fish in both the EE2-H and EE2-M groups. Co-expression analysis of mRNA and lncRNA revealed that EE2 downregulated the transcription of key genes involved in the cell cycle, DNA replication, and Fanconi anemia signaling pathways. Additionally, metabolomic analysis indicated that EE2 influenced metabolism and development-related signaling pathways. These pathways were also significantly identified based on the genes regulated by lncRNA. Consequently, EE2 induced organ deformities and mortality in juvenile zebrafish by disrupting signaling pathways associated with development and metabolism. The results of this study offer new mechanistic insights into the adverse effects of EE2 on juvenile zebrafish based on multiomics analysis. The juvenile zebrafish are highly sensitive to EE2 exposure, which is not limited to adult and embryonic stages. It is a potential model for studying developmental toxicity.

Nickel‐Catalyzed [2+2+2] Cyclotrimerization of Alkynes and Other Unsaturated Systems: A Literature Overview

Transition metal‐catalyzed [2+2+2] cyclotrimerization provides easy access to carbocycles and heterocycles in a single operation with complete atom economy. In recent years, nickel catalysis has emerged as a powerful tool for constructing fused arenes and heteroarenes with high regio‐/chemoselectivity and functional group tolerance. This review highlights the historical perspective and advances made in recent years in nickel‐catalyzed [2+2+2] cycloadditions of alkynes, diynes, and various unsaturated systems. The catalytic activity of the nickel complexes, such as Ni‐phosphine, Ni‐NHC, Ni‐pincer, Ni‐diimine, etc, and their mechanistic insights have been thoroughly examined. Highly substituted arenes have broad applications in pharmaceuticals and soft materials. The applications of cyclotrimerization in these areas have been illustrated.

Selective Protein Degradation through Tetrazine Ligation of Genetically Incorporated Unnatural Amino Acids.

Small molecule-responsive tags for targeted protein degradation are valuable tools for fundamental research and drug target validation. Here, we show that genetically incorporated unnatural amino acids bearing a strained alkene or alkyne functionality can act as a minimalist tag for targeted protein degradation. Specifically, we observed the degradation of strained alkene- or alkyne-containing kinases and E2 ubiquitin-conjugating enzymes upon treatment with hydrophobic tetrazine conjugates. The extent of the induced protein degradation depends on the identity of the target protein, unnatural amino acid, and tetrazine conjugate, as well as the site of the unnatural amino acid in the target protein. Mechanistic studies revealed proteins undergo proteasomal degradation after tetrazine tethering, and the identity of tetrazine conjugates influences the dependence of ubiquitination on protein degradation. This work provides an alternative approach for targeted protein degradation and mechanistic insight, facilitating the future development of more effective targeted protein degradation strategies.

Mechanistic insights into Retama raetam's anti-proliferative and pro-apoptotic effects in A549 lung cancer cells: targeting PI3K/Akt pathway and ROS production.

Lung cancer, particularly non-small cell lung cancer (NSCLC), is a leading cause of cancer-related deaths worldwide. This study investigates the molecular mechanisms behind the anti-cancer effects of the tropical desert plant Retama raetam (R. raetam) on the A549 NSCLC cell line. The research examined R. raetam's anti-proliferative effects, cytotoxicity, apoptosis, reactive oxygen species (ROS) generation, mitochondrial membrane potential, and cell morphology in NSCLC A549 and L-132 cells. In addition, the influence of R. raetam on DNA fragmentation, apoptotic signaling, and PI3K/Akt pathways for its anti-cancer mechanism was examined. Our results indicated that R. raetam's effects were dose- and time-dependent to exhibit anti-proliferative effects on A549 cells. R. raetam treatment promoted apoptotic cell death cycle arrest, increased apoptotic cells, depolarized the mitochondrial membrane, and induced morphological alterations in cells and nuclei. It also inhibited A549 cell migration (P < 0.05), colonization, and invasiveness. Moreover, the study demonstrated that R. raetam treatment resulted in the upregulation of Bax expression, downregulation of Bcl-2 expression, and apoptotic fragmented DNA in A549 cells. The top five bioactive compounds derived from R. raetam exhibited molecular interactions that inhibit PIK3CA and AKT1. This inhibition leads to an increased frequency of apoptosis and subsequent death of cancer cells. Additionally, R. raetam extract induced an increase in ROS formation and cytochrome c levels, indicating that its toxic effects on A549 cells involve both ROS-dependent cytotoxicity through the disruption of mitochondrial transmembrane potential ΔΨm and ROS-independent cell cycle arrest through downregulation BCL-2, PARP, E-Cadherin, PI3K, and Akt expressions pathways.

Radical-Based Enantioconvergent Reductive Couplings of Racemic Allenes and Aldehydes.

Transition metal-catalyzed radical-based enantioconvergent reactions have become a powerful strategy to synthesize enantiopure compounds from racemic starting materials. However, existing methods primarily address precursors with central chirality, neglecting those with axial chirality. Herein, we describe the enantioconvergent reductive coupling of racemic allenes with aldehydes, facilitated by a photoredox, chromium, and cobalt triple catalysis system. This method selectively affords one product from sixteen possible regio- and stereoisomers. The protocol leverages CoIII-H mediated hydrogen atom transfer (MHAT) and Cr-catalyzed radical-polar crossover for efficient stereoablation of axial chirality and asymmetric addition, respectively. Supported by mechanistic insights from control experiments, deuterium labeling, and DFT calculations, our approach offers synthetic chemists a valuable tool for creating enantioenriched chiral homoallylic alcohols, promising to advance radical-based strategies for synthesizing complex chiral molecules.

Synergistic Photoelectrochemical Detection of Cd2+ with Carbon-Bismuth Oxybromide Composites: Mechanistic Insights from DFT Calculations

The existences of hazardous heavy metals like cadmium (Cd) in water bodies necessitates rapid, accurate on-site detection strategies. Herein, a novel photoelectrochemical scheme for sensitive detection of Cd2+ was proposed based on the hybrid carbon-bismuth oxybromide materials (C/Bi4O5Br2 and CNTs/BiOBr). The effects of ultraviolet light (UV-light) illumination during the Cd2+ deposition and desorption stages were analyzed, revealing enhanced photocurrent responses and detection sensitivity compared with those non-illumiantion. The optimal sensitivity of 0.42079 μA/ppb (C/Bi4O5Br2 under both deposition and desorption stages) and 0.15634 μA/ppb (CNTs/BiOBr under desorption stage only) were observed with UV illumination within a Cd2+ range of 10-100 ppb. Density functional theory calculation elucidated that Cd2+ adsorption on the semiconductor surface reduced the badgaps of Bi4O5Br2 and BiOBr, which would benefit electron transfer and redox reactions. Additionally, the interaction between Cd and the semiconductor produced a Schottky barrier, which prolonged the lifetime of the photo-generated electron-hole pairs, where the electrons migrated on the semiconductor surface formed electron capture traps and induced band bending, therefore facilitated the electrons in the conduction band to react with Cd2+ deposited on the electrode surface. The study provides mechanistic insights into the photoelectrochemical processes in enhancng precise and sensitive on-site heavy metal ions detection.

Posttransplant complications: molecular mechanisms and therapeutic interventions.

Posttransplantation complications pose a major challenge to the long-term survival and quality of life of organ transplant recipients. These complications encompass immune-mediated complications, infectious complications, metabolic complications, and malignancies, with each type influenced by various risk factors and pathological mechanisms. The molecular mechanisms underlying posttransplantation complications involve a complex interplay of immunological, metabolic, and oncogenic processes, including innate and adaptive immune activation, immunosuppressant side effects, and viral reactivation. Here, we provide a comprehensive overview of the clinical features, risk factors, and molecular mechanisms of major posttransplantation complications. We systematically summarize the current understanding of the immunological basis of allograft rejection and graft-versus-host disease, the metabolic dysregulation associated with immunosuppressive agents, and the role of oncogenic viruses in posttransplantation malignancies. Furthermore, we discuss potential prevention and intervention strategies based on these mechanistic insights, highlighting the importance of optimizing immunosuppressive regimens, enhancing infection prophylaxis, and implementing targeted therapies. We also emphasize the need for future research to develop individualized complication control strategies under the guidance of precision medicine, ultimately improving the prognosis and quality of life of transplant recipients.

P06-21 Multiparametric in vitro assays combined with high-throughput RNA sequencing for cardiotoxicity risk assessment and mechanistic insight

P06-21 Multiparametric in vitro assays combined with high-throughput RNA sequencing for cardiotoxicity risk assessment and mechanistic insight

Enhanced hydrolytic acidification with Zero-Valent Iron for efficient treatment of comprehensive wastewater from industrial parks: mechanistic insights and toxicity reduction

Enhanced hydrolytic acidification with Zero-Valent Iron for efficient treatment of comprehensive wastewater from industrial parks: mechanistic insights and toxicity reduction

Cardiac Arrhythmias in Pulmonary Arterial Hypertension and Chronic Thromboembolic Pulmonary Hypertension: Mechanistic Insights, Pathophysiology, and Outcomes.

Arrhythmias are increasingly recognized as severe complications of precapillary pulmonary hypertension, encompassing pulmonary arterial hypertension (PAH) and chronic thromboembolic pulmonary hypertension (CTEPH). Despite their significant contribution to symptoms, morbidity, in-hospital mortality, and potentially sudden death in PAH/CTEPH, there remains a lack of comprehensive data on epidemiology, pathophysiology, and outcomes to inform the management of these patients. This review provides an overview of the latest evidence on this subject, spanning from the molecular mechanisms underlying arrhythmias in the hypertrophied or failing right heart to the clinical aspects of epidemiology, diagnosis, and treatment.

Reduction of circulating IgE and allergens by a pH-sensitive antibody with enhanced FcγRIIb binding

Allergen-crosslinked IgE triggers allergy by interacting with its receptor on basophils and mast cells. The anti-IgE monoclonal antibody omalizumab can alleviate allergy by competing with the receptor for IgE binding. However, along with neutralization, omalizumab also inhibits IgE degradation, which is clinically associated with high dose and total IgE accumulation problems. In this study, we have developed an IgE-eliminating antibody on the basis of omalizumab, which has pH-dependent Fabs and an Fc with high affinity for FcγRIIb. In mice, the antibody rapidly eliminated total serum IgE to baseline levels and caused lower free IgE levels than omalizumab. At low dosages, the antibody also exhibited favorable IgE elimination effects. In addition, the antibody can degrade the corresponding allergen with the removal of IgE, addressing the allergy from its source. Introduction of the M252Y/S254T/T256E (YTE) mutation into this antibody prolongs its serum half-life without reducing potency. Thus, this engineered antibody holds a promising therapeutic option for allergy patients. Mechanistic insights are also included in this study.

Mechanistic insights into ultrafast degradation of electron-rich emerging pollutants by waste cyanobacteria resource utilization

Biochar has the issues with inadequate stability and low catalytic activity. In this study, two-step ball milling method was applied to synthesis carbon nitride co-modified cyanobacteria biochar (CN-BC). The optimized CN-BC can effectively activate peroxymonosulfate (PMS) to degrade emerging pollutants. The degradation rate constant of enrofloxacin in the CN-BC/PMS system reached 0.506 min−1, and 95 % of enrofloxacin could be effectively degraded within 10 min. Free radical quenching experiments and electrochemical experiments confirmed that the electron transfer and 1O2 were the main pathways in the CN-BC/PMS system. The CN-BC/PMS system has selective degradation of electron-rich pollutants, and the pollutants containing electron-donating groups are more easily degraded than those containing electron-withdrawing group. The selective degradation pathway of pollutants by CN-BC was further analyzed using density functional theory (DFT). Continuous flow CN-BC/PMS membrane system showed long-term abatement of emerging pollutants. This study improves the understanding of the mechanism of PMS activation by cyanobacterial biochar catalysts and provides a new pathway for membrane-based domain-limited catalysis.