Transgenic, high-protein sorghums display promise in poultry diets in an initial comparison.

This study aimed to compare the inclusion of transgenic sorghums against commercially available sorghums on growth performance in broiler chickens. Isonitrogenous and isoenergetic diets were offered to a total 288 male Ross 308 broiler chickens from 14 to 35 d posthatch. Three dietary treatments were diets based on transgenic sorghums with a mean protein content of 154.7 g/kg and 5 treatments were based on commercially available sorghum hybrids with a mean protein content of 90.6 g/kg. Soybean meal inclusions in the commercial sorghum diets averaged 215 g/kg, which was reduced to 171 g/kg in the transgenic sorghum diets because of the higher protein contents. Overall growth performance was highly satisfactory, and commercial sorghums supported 2.55% (2,330 vs. 2,272 g/bird; P = 0.010) more weight gains and 2.74% (2,929 vs. 2,851 g/bird; P = 0.012) higher feed intakes; however, the transgenic sorghums supported a fractionally better FCR (1.255 vs 1.257; P = 0.826). There were no statistical differences in apparent jejunal and ileal starch and protein (N) digestibility coefficients between treatments. The transgenic sorghum diets generated slightly, but significantly, higher AME:GE ratios and AMEn, but the commercial sorghum diets generated 6.33% (235 vs. 221 g/kg; P < 0.001) greater breast meat yields. Apparent ileal digestibility coefficients of 16 amino acids averaged 0.839 and 0.832 for transgenic and commercial sorghum-based diets, respectively, without any significant differences in individual amino acids. This outcome suggests amino acid digestibilities of the transgenic sorghums may be inherently higher than commercial hybrid sorghums as the 25.7% higher average soybean meal inclusions would have advantaged amino acid digestibilities in commercial sorghum diets. The possibility that the digestibilities of amino acids in the kafirin component of transgenic sorghums was enhanced by modifications to the structure of kafirin protein bodies is discussed. In conclusion, transgenic sorghums with higher protein concentrations led to 20.5% reduction of soybean meal inclusions in broiler diets, and this change did not compromise feed conversion efficiency compared to standard commercial hybrid sorghums.

LAMP1 controls CXCL10-CXCR3 axis mediated inflammatory regulation of macrophage polarization during inflammatory stimulation.

Increased expression of CXCL10 and its receptor CXCR3 represents an inflammatory response in cells and tissues. Macrophage polarization and autophagy are major functions in inflammatory macrophages; however, the cellular functions of the CXCL10-CXCR3 axis in macrophages are not well understood. Here, we examined the role of CXCL10-CXCR3-axis-regulated autophagy in macrophage polarization. First, in non-inflammatory macrophages, whereas CXCL10 promotes M2 polarization and inhibits M1 polarization, CXCR3 antagonist AMG487 induces the opposite macrophage polarization. Next, CXCL10 promotes the expression of autophagy proteins (Atg5-Atg12 complex, p62, LC3-II, and LAMP1) and AMG487 inhibits their expression. Knockdown of LAMP1 by short interfering RNA switches the CXCL10-induced polarization from M2 to M1 in non-inflammatory macrophages. Furthermore, in inflammatory macrophages stimulated by poly(I:C), CXCL10 induces M1 polarization and AMG487 induces M2 polarization in association with a decrease in LAMP1. Finally, AMG487 alleviates lung injury after poly(I:C) treatment in mice. In conclusion, CXCL10-CXCR3 axis differentially directs macrophage polarization in inflammatory and non-inflammatory states, and autophagy protein LAMP1 acts as the switch controlling the direction of macrophage polarization by CXCL10-CXCR3.

Effect of faba bean Vicia faba L. water/alcohol extract on growth performance, antioxidant capacity, textural properties, and collagen deposition in the swim bladder of juvenile Nibea coibor.

Faba bean has gained attention as a cost-effective protein source with the potential to enhance product quality (texture properties, collagen content, etc.) in fish. However, its anti-nutrition factor, high feed conversion ratio, poor growth performance, etc. limit the widely application as a dietary source, especially in carnivorous fish. The water or alcohol extract of faba bean might resolve the problem. In this study, the juvenile Nibea coibor, known for their high-protein, large-sized, and high-grade swim bladder, were fed with seven isoproteic and isolipid experimental diets with the additive of faba bean water extract (1.25%, 2.5%, and 5%) or faba bean alcohol extract (0.9%, 1.8%, and 3.6%), with a control group without faba bean extract. After the 10-week feeding trail, the growth, antioxidant capacity, textural properties, and collagen deposition of the swim bladder were analyzed. Results showed that the 1.25% faba bean water extract group could significantly promote growth, textural quality of the swim bladder, and have beneficial effects on antioxidant response of fish. Conversely, dietary supplementation of faba bean alcohol extract resulted in reduced growth performance in a dose-dependent manner. Furthermore, fish fed diet with 1.25% faba bean water extract exhibited increased collagen content and upregulated collagen-related gene expression in the swim bladder, which was consistent with the Masson stain analysis for collagen fiber. Our results suggested that the anti-nutrient factor and bioactive component of faba bean may mainly be enriched in alcohol extract and water extract of faba bean, respectively. Besides, the appropriate addition of water extract of faba bean may improve the texture quality of the swim bladder by promoting collagen deposition. This study would provide a theoretical basis for the formulated diets with faba bean extract to promote product quality of marine fish.

Assembly and Utility of a Drawstring-Mimetic Supramolecular Complex.

Inspired by the drawstring structure in daily life, here we report the development of a drawstring-mimetic supramolecular complex at the molecular scale. This complex consists of a rigid figure-of-eight macrocyclic host molecule and a flexible linear guest molecule which could interact through three-point non-covalent binding to form a highly selective and efficient host-guest assembly. The complex not only resembles the drawstring structure, but also mimics the properties of a drawstring with regard to deformations under external forces. The supramolecular drawstring can be utilized as an interlocked crosslinker for poly(methyl acrylate), and the corresponding polymer samples exhibit comprehensive enhancement of macroscopic mechanical performance including stiffness, strength, and toughness.

ACE2 improves endothelial cell function and reduces acute lung injury by downregulating FAK expression.

Endothelial cell (EC) barrier dysfunction and increased adhesion of immune inflammatory cells to ECs crucially contribute to acute lung injury (ALI). Angiotensin-converting enzyme 2 (ACE2) is an essential regulator of the renin-angiotensin system (RAS) and exerts characteristic vasodilatory and anti-inflammatory effects. SARS-COV-2 infects the lungs by binding to ACE2, which can lead to dysregulation of ACE2 expression, further leading to ALI with predominantly vascular inflammation and eventually to more severe acute respiratory distress syndrome (ARDS). Therefore, restoration of ACE2 expression represents a valuable therapeutic approach for SARS-COV-2-related ALI/ARDS. In this study, we used polyinosinic-polycytidylic acid (Poly(I:C)), a double-stranded RNA analog, to construct a mouse ALI model that mimics virus infection. After Poly(I:C) exposure, ACE2 was downregulated in mouse lung tissues and in cultured ECs. Treatment with DIZE, an ACE2-activating compound, upregulated ACE2 expression and relieved ALI in mice. DIZE also improved barrier function and reduced the number of THP-1 monocytes adhering to cultured ECs. Focal adhesion kinase (FAK) and phosphorylated FAK (p-FAK) levels were increased in lung tissues of ALI mice as well as in Poly(I:C)-treated ECs in vitro. Both DIZE and the FAK inhibitor PF562271 decreased FAK/p-FAK expression in both ALI models, attenuating ALI severity in vivo and increasing barrier function and reducing monocyte adhesion in cultured ECs. Furthermore, in vivo experiments using ANG 1-7 and the MAS inhibitor A779 corroborated that DIZE-mediated ACE2 activation stimulated the activity of the ANG 1-7/MAS axis, which inhibited FAK/p-FAK expression in the mouse lung. These findings provide further evidence that activation of ACE2 in ECs may be a valuable therapeutic strategy for ALI.

DNMT3B Alleviates Liver Steatosis Induced by Chronic Low-grade LPS via Inhibiting CIDEA Expression.

BACKGROUND & AIMS: Nonalcoholic fatty liver disease is the most prevalent chronic liver disease and threats to human health. Gut dysbiosis caused by lipopolysaccharide (LPS) leakage has been strongly related to nonalcoholic fatty liver disease progression, although the underlying mechanisms remain unclear. METHODS: Previous studies have shown that low-grade LPS administration to mice on a standard, low-fat chow diet is sufficient to induce symptoms of fatty liver. This study confirmed these findings and supported LPS as a lipid metabolism regulator in the liver. RESULTS: Mechanically, LPS induced dysregulated lipid metabolism by inhibiting the expression of DNA methyltransferases 3B (DNMT3B). Genetic overexpression of DNMT3B alleviated LPS-induced lipid accumulation, whereas its knockdown increased steatosis in mice and human hepatocytes. LPS-induced lower expression of DNMT3B led to hypomethylation in promoter region of CIDEA, resulting in increased binding of SREBP-1c to its promoter and activated CIDEA expression. Hepatic interference of CIDEA reversed the effect of LPS on lipogenesis. These effects were independent of a high-fat diet or high fatty acid action. CONCLUSIONS: Overall, these findings sustain the conclusion that LPS is a lipogenic factor and could be involved in hepatic steatosis progression.

Enhancement of Pt-O Synergistic Sites through Titanium Vacancies for Low-Temperature Nitrogen Oxide Reduction.

Improving the reaction rate of each step is significant for accelerating the multistep reaction of NO reduction by H2. However, simultaneously enhancing the activation of different gaseous reactants using single-atom catalysts remains a challenge to maximize the activity. Herein, we propose a strategy that utilizes titanium-vacancy-regulated electronic properties of single atoms and defective support (Pt1/d-TiO2) to facilitate electron transfer from edge-share O atoms (OTi) to adjacent Pt single atoms. This leads to the formation of low-valence Pt and unsaturated-charge OTi sites, which causes the catalytic reaction to follow a synergistic mechanism. Specifically, experimental and theoretical analyses demonstrate that low-valence Pt sites finely tune the adsorption of H2 molecules, consequently lowering the dissociation energy from 0.15 to as low as 0.01 eV. Moreover, using quasi-in situ spectroscopy, we clearly observe NO molecules being adsorbed on interfacial oxygen sites of a defective support. Then, the bond energy of the N-O bond is weakened through an electron acceptance-donation mechanism between unsaturated-charge OTi sites and NO, thereby facilitating NO activation. The designed single-atom catalysts with synergistic sites exhibit unmatched activity at low temperatures (above 90% NOx conversion at 100 °C), along with higher turnover frequency value (0.74 s-1) and superior stability, making them potentially suitable for industrial applications.

Optical fiber flow sensor based on a lever-hinge configuration.

Based on a lever-hinge structure, a target-type fiber Bragg grating (FBG) flow sensor is proposed. Differential measurements of temperature and pressure are achieved using two FBGs. The design idea of the sensor is demonstrated from both theoretical and experimental aspects, and the relationship between FBG wavelength and temperature and the relationship between FBG wavelength and volume flow rate were established, respectively. The sensor is compact with good resolution, high stability, wide measurement range, and easy fabrication, and can be applied to measure temperature and volume flow rate in injection wells.

Insights into the Correlation of Cross-linking Modes with Mechanical Properties for Dynamic Polymeric Networks.

Simultaneously introducing covalent and supramolecular cross-links into one system to construct dually cross-linked networks, has been proved an effective approach to prepare high-performance materials. However, so far, features and advantages of dually cross-linked networks compared with those possessing individual covalent or supramolecular cross-linking points are rarely investigated. Herein, on the basis of comparison between supramolecular polymer network (SPN), covalent polymer network (CPN) and dually cross-linked polymer network (DPN), we reveal that the dual cross-linking strategy can endow the DPN with integrated advantages of CPN and SPN. Benefiting from the energy dissipative ability along with the dissociation of host-guest complexes, the DPN shows excellent toughness and ductility similar to the SPN. Meanwhile, the elasticity of covalent cross-links in the DPN could rise the structural stability to a level comparable to the CPN, exhibiting quick deformation recovery capacity. Moreover, the DPN has the strongest breaking stress and puncture resistance among the three, proving the unique property advantages of dual cross-linking method. These findings gained from our study further deepen the understanding of dynamic polymeric networks and facilitate the preparation of high-performance elastomeric materials.

Regioselective stilbene O-methylations in Saccharinae grasses.

O-Methylated stilbenes are prominent nutraceuticals but rarely produced by crops. Here, the inherent ability of two Saccharinae grasses to produce regioselectively O-methylated stilbenes is reported. A stilbene O-methyltransferase, SbSOMT, is first shown to be indispensable for pathogen-inducible pterostilbene (3,5-bis-O-methylated) biosynthesis in sorghum (Sorghum bicolor). Phylogenetic analysis indicates the recruitment of genus-specific SOMTs from canonical caffeic acid O-methyltransferases (COMTs) after the divergence of Sorghum spp. from Saccharum spp. In recombinant enzyme assays, SbSOMT and COMTs regioselectively catalyze O-methylation of stilbene A-ring and B-ring respectively. Subsequently, SOMT-stilbene crystal structures are presented. Whilst SbSOMT shows global structural resemblance to SbCOMT, molecular characterizations illustrate two hydrophobic residues (Ile144/Phe337) crucial for substrate binding orientation leading to 3,5-bis-O-methylations in the A-ring. In contrast, the equivalent residues (Asn128/Asn323) in SbCOMT facilitate an opposite orientation that favors 3'-O-methylation in the B-ring. Consistently, a highly-conserved COMT is likely involved in isorhapontigenin (3'-O-methylated) formation in wounded wild sugarcane (Saccharum spontaneum). Altogether, our work reveals the potential of Saccharinae grasses as a source of O-methylated stilbenes, and rationalize the regioselectivity of SOMT activities for bioengineering of O-methylated stilbenes.

Discovery of ML210-Based glutathione peroxidase 4 (GPX4) degrader inducing ferroptosis of human cancer cells.

Ferroptosis is an iron-dependent cell death caused by the accumulation of lipid peroxidation. The glutathione peroxidase 4 (GPX4) is an antioxidative enzyme and a major regulator of ferroptosis. Targeting GPX4 has become a promising strategy for cancer therapy. Here in this article, we designed and synthesized a series of GPX4 degraders using ML210 as a warhead. DC-2 among them has been found to have the best degradation activity with the DC50 value of 0.03 µM in HT1080 cells. It also showed an obvious cell growth inhibition effect with the IC50 value of 0.1 µM in HT1080 cells. Mechanism research showed that DC-2 induced GPX4 degradation via the ubiquitin-proteasome pathway and autophagy-lysosome pathway. GPX4 degradation induced by DC-2 could result in the accumulation of ROS and subsequent ferroptosis. The pharmacodynamics study showed that DC-2 could reduce the GPX4 level in HT1080 tumor tissue in mice and has a good safety profile. Above all, a potent and safe compound DC-2 has been found to induce GPX4 degradation and subsequent ferroptosis. This study may lay the foundation for a highly efficient and safe drug with a new mechanism for cancer therapy.

Structure-activity relationship study of RSL3-based GPX4 degraders and its potential noncovalent optimization.

Ferroptosis is an iron-dependent, non-apoptotic form of cell death involving in various disease processes. Mechanistically, glutathione peroxidase 4 (GPX4) which belongs to the redox enzyme can convert lipid hydroperoxides into innocuous lipid alcohol to protect cells from ferroptosis. Therefore, targeting manipulation of GPX4 may represent a promising strategy for regulating cell redox homeostasis and ferroptosis. In this work, we designed, synthesized and evaluated a series of RSL3-based GPX4 degraders using PROTAC strategy. The structure-activity relationship of these compounds with different E3 ligase ligands, linker lengths and chemical compositions was systematically studied. Compound R17 with carbon chain linker and lenalidomide E3 ligand was selected as the most potent GPX4 degrader for degrading GPX4 protein in nanomolar level either in wild tumor cells or in drug-resistant tumor cells. We also optimized the POI ligand of R17 with chloracetylamine replaced to propionamide to construct noncovalent GPX4 degrader NC-R17. Such noncovalent modification led to a moderate GPX4 degradation activity and represents a promising strategy for the development of noncovalent GPX4 PROTACs. In general, we screened a set of GPX4 degraders to give the compound R17 with excellent protein degradation activity, and further optimization gave the noncovalent degrader NC-R17 with moderate efficacy. These results lay a firm foundation for the discovery of novel anti-tumor drugs targeting GPX4 and offer the proof of concept for the design of noncovalent GPX4 PROTACs.

Oligo[2]catenane That Is Robust at Both the Microscopic and Macroscopic Scales.

Polycatenanes are extremely attractive topological architectures on account of their high degrees of conformational freedom and multiple motion patterns of the mechanically interlocked macrocycles. However, exploitation of these peculiar structural and dynamic characteristics to develop robust catenane materials is still a challenging goal. Herein, we synthesize an oligo[2]catenane that showcases mechanically robust properties at both the microscopic and macroscopic scales. The key feature of the structural design is controlling the force-bearing points on the metal-coordinated core of the [2]catenane moiety that is able to maximize the energy dissipation of the oligo[2]catenane via dissociation of metal-coordination bonds and then activation of sequential intramolecular motions of circumrotation, translation, and elongation under an external force. As such, at the microscopic level, the single-molecule force spectroscopy measurement exhibits that the force to rupture dynamic bonds in the oligo[2]catenane reaches a record high of 588 ± 233 pN. At the macroscopic level, our oligo[2]catenane manifests itself as the toughest catenane material ever reported (15.2 vs 2.43 MJ/m3). These fundamental findings not only deepen the understanding of the structure-property relationship of poly[2]catenanes with a full set of dynamic features but also provide a guiding principle to fabricate high-performance mechanically interlocked catenane materials.

Multiphase holdup measurement of oil-gas-water flow using new dual receiver fiber-optical probe array multiphase logging tool.

To solve the problem of multiphase holdup measurement, a new dual-receiver fiber-optical probe array multiphase logging tool (NDRFOPA_MLT) is designed and developed. This paper first constructed the mechanism model of an NDRFOP for phase holdup measurement by using the ray tracing method and theoretically analyzed the feasibility of NDRFOP to measure phase holdup; considering the shortcomings of NDRFOP local measurement, a NDRFOPA sensor for oil production three-phase flow is designed and developed. At the same time, the volume of fluid model was used to simulate the distribution characteristics of the medium in the NDRFOPA_MLT measurement pipeline under the working conditions of oil-gas-water flow with a total flow rate range of 0.42-1.25 m3/h, water holdup range of 50%-80%, oil holdup range of 10%-30%, and gas holdup range of 10%-40%. In addition, the NDRFOPA_MLT measurement models for different multiphase flow conditions were established by the ZEMAX ray tracing method, and the sensitivity distribution, response characteristics, and phase holdup measurement methods were studied to obtain the phase holdup measurement results under multiphase flow conditions. Finally, a multiphase flow experimental platform with a measurement pipe diameter of 20 mm and a measurement pipe length of 300 mm was established, and experiments were conducted under multiphase flow conditions, such as a gas flow rate range of 0.04-0.16 m3/h, oil flow rate range of 0.64-1.70 m3/h, and water flow rate range of 0.53-2.58 m3/h. The experimental results showed that phase holdup measurement error was mainly kept within 10%.

Accumulated cholesterol protects tumours from elevated lipid peroxidation in the microenvironment.

Elevated lipid peroxidation (LPO), usually present in the tumour microenvironment (TME), is profoundly implicated in antitumour immunity and may be targeted for the development of new antitumour therapies. However, tumour cells may also rewire their metabolism to survive elevated LPO. Here, we report a novel and nonantioxidant mechanism by which tumour cells benefit from accumulated cholesterol to restrain LPO and ferroptosis, a nonapoptotic form of cell death characterized by accumulated LPO. Modulating cholesterol metabolism, especially LDLR-mediated cholesterol uptake, shifted the susceptibility of tumour cells to ferroptosis. Elevation of cellular cholesterol content specifically restrained LPO triggered by GSH-GPX4 inhibition or oxidizing factors in the TME. Furthermore, depletion of TME cholesterol by MßCD efficiently enhanced the antitumour efficacy of ferroptosis in a mouse xenograft model. Distinct from the antioxidant effect of its metabolic intermediates, the protective role of cholesterol was ascribed to its ability to decrease membrane fluidity and promote lipid raft formation, which affects the diffusion of LPO substrates. A correlation between LPO and lipid rafts was also found in tumour tissues from renal cancer patients. Together, our findings have identified a general and nonsacrificial mechanism by which cholesterol suppresses LPO, which can be exploited to enhance the efficacy of ferroptosis-based antitumour strategies.

In situ formation of cocatalytic sites boosts single-atom catalysts for nitrogen oxide reduction.

Nitrogen oxide (NOx) pollution presents a severe threat to the environment and human health. Catalytic reduction of NOx with H2 using single-atom catalysts poses considerable potential in the remediation of air pollution; however, the unfavorable process of H2 dissociation limits its practical application. Herein, we report that the in situ formation of PtTi cocatalytic sites (which are stabilized by Pt-Ti bonds) over Pt1/TiO2 significantly increases NOx conversion by reducing the energy barrier of H2 activation. We demonstrate that two H atoms of H2 molecule are absorbed by adjacent Pt atoms in Pt-O and Pt-Ti, respectively, which can promote the cleave of H-H bonds. Besides, PtTi sites facilitate the adsorption of NO molecules and further lower the activation barrier of the whole de-NOx reaction. Extending the concept to Pt1/Nb2O5 and Pd1/TiO2 systems also sees enhanced catalytic activities, demonstrating that engineering the cocatalytic sites can be a general strategy for the design of high-efficiency catalysts that can benefit environmental sustainability.

Dual degradation mechanism of GPX4 degrader in induction of ferroptosis exerting anti-resistant tumor effect.

Targeting Glutathione peroxidase 4 (GPX4) has become a promising strategy for drug-resistant cancer therapy via ferroptosis induction. It was found that the GPX4 inhibitors such as RSL3 have GPX4 degradation ability via not only autophagy-lysosome pathway but also ubiquitin-proteasome system (UPS). Proteolysis targeting chimeras (PROTACs) using small molecule with both inhibition and degradation ability as the ligand of protein of interest (POI) have not been reported. To obtain better compounds with effective disturbance of GPX4 activity, and compare the difference between GPX4 inhibitors with degradation ability and their related PROTACs, we designed and synthesized a series of GPX4 degraders using PROTAC technology in terms of its excellent characteristics such as high efficiency and selectivity and the capacity of overcoming resistance. Hence, 8e was discovered as a potent and highly efficacious GPX4 degrader based upon the inhibitor RSL3. It was 2-3 times more potent than RSL3 in all the in vitro anti-tumor assays, indicating the importance of the PROTAC ternary complex of GPX4, 8e and E3 ligase ligand. 8e revealed better potency in resistant tumor cells than in wide type cells. Furthermore, we discovered for the first time that degrader 8e exhibit GPX4 degradation activity via ubiquitin-proteasome system (UPS) and autophagy-lysosome pathway with UPS plays the major role in the process. Our data also suggested that 8e and RSL3 could potently induce ferroptosis of HT1080 cells via GPX4 inhibition and degradation. In summary, our data revealed that the GPX4 degrader 8e achieves better degradation and anti-tumor effects compared to its related GPX4 inhibitor RSL3. Thus, an efficient strategy to induce GPX4 degradation and subsequent ferroptosis was established in this study for malignant cancer treatment in the future.

Bromide-Mediated Photoelectrochemical Epoxidation of Alkenes Using Water as an Oxygen Source with Conversion Efficiency and Selectivity up to 100.

In a photoelectrochemical (PEC) cell, the production of solar fuels such as hydrogen is often accompanied either by the oxidation of water or by the oxidation of organic substrates. In this study, we report bromide-mediated PEC oxidation of alkenes at a mesoporous BiVO4 photoanode and simultaneous hydrogen evolution at the cathode using water as an oxygen source. NaBr as a redox mediator was demonstrated to play a dual role in the PEC organic synthesis, which facilitates the selective oxidation of alkenes into epoxides and suppresses the photocorrosion of BiVO4 in water. This method enables a near-quantitative yield and 100% selectivity for the conversion of water-soluble alkenes into their epoxides in H2O/CH3CN solution (v/v, 4/1) under simulated sunlight without the use of noble metal-containing catalysts or toxic oxidants. The maximum solar-to-electricity efficiency of 0.58% was obtained at 0.39 V vs Ag/AgCl. The obtained epoxide products such as glycidol are important building blocks of the chemical industry. Our results provide an energy-saving and environment-benign approach for producing value-added chemicals coupled with solar fuel generation.

Phosphorothioate-Based Site-Specific Labeling of Large RNAs for Structural and Dynamic Studies.

Pulsed electron-electron double resonance (PELDOR) spectroscopy, X-ray scattering interferometry (XSI), and single-molecule Förster resonance energy transfer (smFRET) are molecular rulers that provide inter- or intramolecular pair-wise distance distributions in the nanometer range, thus being ideally suitable for structural and dynamic studies of biomolecules including RNAs. The prerequisite for such applications requires site-specific labeling of biomolecules with spin labels, gold nanoparticles, and fluorescent tags, respectively. Recently, site-specific labeling of large RNAs has been achieved by a combination of transcription of an expanded genetic alphabet containing A-T/G-C base pairs and NaM-TPT3 unnatural base pair (UBP) with post-transcriptional modifications at UBP bases by click chemistry or amine-NHS ester reactions. However, due to the bulky sizes of functional groups or labeling probes used, such strategies might cause structural perturbation and decrease the accuracy of distance measurements. Here, we synthesize an α-thiophosphorylated variant of rTPT3TP (rTPT3αS), which allows for post-transcriptional site-specific labeling of large RNAs at the internal α-phosphate backbone via maleimide-modified probes. Subsequent PELDOR, XSI, and smFRET measurements result in narrower distance distributions than labeling at the TPT3 base. The presented strategy provides a new route to empower the molecular rulers for structural and dynamic studies of large RNA and its complex.

PDTAC: Targeted Photodegradation of GPX4 Triggers Ferroptosis and Potent Antitumor Immunity.

Targeted degradation of proteins, especially those regarded as undruggable or difficult to drug, attracts wide attention to develop novel therapeutic strategies. Glutathione peroxidase 4 (GPX4), the key enzyme regulating ferroptosis, is currently a target with just covalent inhibitors. Here, we developed a targeted photolysis approach and achieved efficient degradation of GPX4. The photodegradation-targeting chimeras (PDTACs) were synthesized by conjugating a clinically approved photosensitizer (verteporfin) to noninhibitory GPX4-targeting peptides. These chimeras selectively degraded the target protein in both cell lysates and living cells upon red-light irradiation. The targeted photolysis of GPX4 resulted in dominant ferroptotic cell death in malignant cancer cells. Moreover, the dying cells resulting from the PDTACs exhibited potent immunogenicity in vitro and efficiently elicited antitumor immunity in vivo. Our approach therefore provides a novel method to induce GPX4 dysfunction based on noncovalent binding and specifically trigger immunogenic ferroptosis, which may boost the application of ferroptosis in cancer immunotherapy.