Stimulating Phonon Bottleneck Effect in Organic Semiconductors by Charge-Transfer-Mediated J-Aggregation.

Hot carriers rapidly lose kinetic energies on a subpicosecond time scale, posing significant limitations on semiconductors' photon-conversion efficiencies. To slow the hot carrier cooling, the phonon bottleneck effect is constructed prevalently in quantum-confined structures with discrete energy levels. However, the maximum energy separation (ΔEES) between the energy levels is in a range of several hundred meV, leading to unsatisfactory cooling time. To address this, we design a novel organic semiconductor capable of forming intermolecular charge transfer (CT) in J-aggregates, where the lowest singlet excited state (S1) splits into two states due to the significant interplay between the Coulomb interaction and intermolecular CT coupling. The ΔEES between the two states can be adjusted up to 1.02 eV, and an extremely slow carrier cooling process of ∼72.3 ps was observed by femtosecond transient absorption spectroscopy. Moreover, the phonon bottleneck effect was identified in organic materials for the first time, and CT-mediated J-aggregation with short-range interactions was found to be the key to achieving large ΔEES. The significantly prolonged carrier cooling time, compared to <100 fs in the isolated molecule (10-6 M), highlights the potential of organic molecules with diversified aggregation structures in achieving long-lived hot carriers. These findings provide valuable insights into the intrinsic photophysics of electron-phonon scattering in organic semiconductors.

PDZK1 improves ventricular remodeling in hypertensive rats by regulating the stability of the Mas receptor.

Ventricular remodeling is one of the main causes of mortality from heart failure due to hypertension. Exploring its mechanism and finding therapeutic targets have become urgent scientific problems to be solved. A number of studies have shown that Mas, as an Ang-(1-7) specific receptor, was significantly reduced in myocardial tissue of rats undergoing hypertensive ventricular remodeling. It has been reported that Mas receptor levels are significantly downregulated in myocardium undergoing ventricular remodeling, but studies focused on intracellular and post-translational modifications of Mas are lacking. The results of this research are as follows: (1) PDZK1 interacts with the carboxyl terminus of Mas through its PDZ1 domain; (2) the expression of PDZK1 and Mas is decreased in rats undergoing hypertensive ventricular remodeling, and PDZK1 upregulation can ameliorate hypertensive myocardial fibrosis and myocardial hypertrophy; (3) PDZK1 enhances the stability of Mas protein through the proteasome pathway, and the proteasome inhibitor MG132 promotes hypertensive ventricular remodeling. PDZK1 improves ventricular remodeling in hypertensive rats by regulating Mas receptor stability. This study provides a scientific basis for the prevention and treatment of ventricular remodeling.

Effect of different vegetation restoration on soil organic carbon dynamics and fractions in the Rainy Zone of Western China.

Vegetation restoration on purple soil (Eutric Leptic Regosols) slopes aiming at reducing soil erosion in the Rainy Zone of Western China has significantly altered soil organic carbon (SOC) storage and distribution. A better understanding of the effects of different vegetation restoration types on SOC dynamics and fractions is critical in devising better policy to protect or enhance SOC stocks to improve soil quality and ecosystem function. In the present study, total, labile, and non-labile organic carbon (TOC, LC, and NLC), and carbon management index (CMI) of Cryptomeria fortunei (CF), mixed C. fortunei and Betula luminifera (MF), Neosinocalamus affinis (NA), and Camellia sinensis (CS) were compared with those of Zea mays field (ZM) on purple soil slopes in the Rainy Zone of Western China in order to develop more effective ways to implement vegetation restoration in the future. Different vegetation restoration types (CF, MF, NA and CS) increased TOC stock by 47.79%-118.31% and NLC stock by 56.61%-129.52% in the 0-50 cm soil layer compared with that of ZM. The direction and magnitude of changes in LC stock and CMI, however, depended strongly on the vegetation restoration type. Compared with ZM, CF had the largest increase of LC stock and CMI, whereas NA had the largest decrease of LC stock and CMI in the 0-50 cm soil layer. The LC:TOC ratio in four reforested species all declined significantly compared with that of ZM (p < 0.01), indicating decreased SOC activity after afforestation. The vegetation type and soil depth together explained more than 90% of the changes of TOC and its fractions in the plantations on purple soil slopes. Our study demonstrates that transforming the ZM into the CS is optimal to achieve the sustainable development goal, whereas transforming the ZM into the NA reduces the SOC activity and availability.

Effects of land use change on soil carbon and nitrogen in purple paddy soil.

Rural land use patterns in southern China centered on household grain crop production have observed significant changes in the past few decades, profoundly affecting the release and fixation of carbon and nitrogen in the paddy soil of the region. This study selected different land use patterns developed in purple paddy soil on a decadal time scale, examined the changing rate of soil carbon and nitrogen of the purple paddy soil after abandonment, dry-farming, and fish-farming, and revealed the impact of land use changes on the balance of soil carbon and nitrogen. Results showed that the loss rates of soil organic carbon, readily oxidizable organic carbon and total nitrogen at the initial stage of dry-farming were most considerable, followed by abandonment and fish-farming. An average of 11.95-13.94 g kg-1 soil organic carbon loss and 0.90-1.03 g kg-1 total nitrogen loss of the cultivation horizon were observed when purple paddy soil was abandoned and dry farmed. In comparison, the net release of soil organic carbon and total nitrogen after fish-farming were 6.64 and -0.23 g kg-1. The changes of land use of rural area driven by rising labor cost and market demand have been inducing a continuous decline in soil C:N and significantly reducing the purple paddy soil's carbon sequestration ability. The promotion of no-tillage management, increase of organic manure application, and avoidance of over-use of nitrogen fertilizer in dryland farming need to be further considered to meet the dual pressures of China's resource constraints and carbon neutrality goals. A regression model may predict the changes in soil carbon after the change of paddy soil utilization, which provides a pathway for predicting changes in farmland carbon sequestration potential and carbon storage caused by changes in paddy soil utilization in the future.

Dietary leucine supplementation alters energy metabolism and induces slow-to-fast transitions in longissimus dorsi muscle of weanling piglets.

Leucine plays an important role in promoting muscle protein synthesis and muscle remodelling. However, what percentage of leucine is appropriate in creep feed and what proteome profile alterations are caused by dietary leucine in the skeletal muscle of piglets remain elusive. In this case, we applied isobaric tags for relative and absolute quantitation to analyse the proteome profile of the longissimus dorsi muscles of weanling piglets fed a normal leucine diet (NL; 1·66 % leucine) and a high-leucine diet (HL; 2·1 % leucine). We identified 157 differentially expressed proteins between these two groups. Bioinformatics analysis of these proteins exhibited the suppression of oxidative phosphorylation and fatty acid ß-oxidation, as well as the activation of glycolysis, in the HL group. For further confirmation, we identified that SDHB, ATP5F1, ACADM and HADHB were significantly down-regulated (P<0·01, except ATP5F1, P<0·05), whereas the glycolytic enzyme pyruvate kinase was significantly up-regulated (P<0·05) in the HL group. We also show that enhanced muscle protein synthesis and the transition from slow-to-fast fibres are altered by leucine. Together, these results indicate that leucine may alter energy metabolism and promote slow-to-fast transitions in the skeletal muscle of weanling piglets.

Global Liver Proteome Analysis Using iTRAQ Reveals AMPK-mTOR-Autophagy Signaling Is Altered by Intrauterine Growth Restriction in Newborn Piglets.

Intrauterine growth restriction (IUGR) impairs fetal growth and development, perturbs nutrient metabolism, and increases the risk of developing diseases in postnatal life. However, the underlying mechanisms by which IUGR affects fetal liver development and metabolism remain incompletely understood. Here, we applied a high-throughput proteomics approach and biochemical analysis to investigate the impact of IUGR on the liver of newborn piglets. As a result, we identified 78 differentially expressed proteins in the three biological replicates, including 31 significantly up-regulated proteins and 47 significantly down-regulated proteins. Among them, a majority of differentially expressed proteins were related to nutrient metabolism and mitochondrial function. Additionally, many significantly down-regulated proteins participated in the mTOR signaling pathway and the phagosome maturation signaling pathway. Further analysis suggested that glucose concentration and hepatic glycogen storage were both reduced in IUGR newborn piglets, which may contribute to AMPK activation and mTORC1 inhibition. However, AMPK activation and mTORC1 inhibition failed to induce autophagy in the liver of IUGR neonatal pigs. A possible reason is that PP2Ac, a potential candidate in autophagy regulation, is significantly down-regulated in the liver of IUGR newborn piglets. These findings may provide implications for preventing and treating IUGR in human beings and domestic animals.

Quantitative proteomics analysis reveals glutamine deprivation activates fatty acid ß-oxidation pathway in HepG2 cells.

Glutamine, a multifunctional amino acid, functions in nutrient metabolism, energy balance, apoptosis, and cell proliferation. Lipid is an important nutrient and controls a broad range of physiological processes. Previous studies have demonstrated that glutamine can affect lipolysis and lipogenesis, but the effect of glutamine on the detailed lipid metabolism remains incompletely understood. Here, we applied the quantitative proteomics approach to estimate the relative abundance of proteins in HepG2 cells treated by glutamine deprivation. The results showed that there were 212 differentially abundant proteins in response to glutamine deprivation, including 150 significantly increased proteins and 62 significantly decreased proteins. Interestingly, functional classification showed that 43 differentially abundant proteins were related to lipid metabolism. Further bioinformatics analysis and western blotting validation revealed that lipid accumulation may be affected by ß-oxidation of fatty acid induced by glutamine deprivation in HepG2 cells. Together, our results may provide the potential for regulating lipid metabolism by glutamine in animal production and human nutrition. The MS data have been deposited to the ProteomeXchange Consortium with identifier PXD003387.

Influence of psychological intervention on pain and immune functions of patients receiving lung cancer surgery.

OBJECTIVE: To observe the influence of psychological intervention on pain, immune system and adrenocortical functions of patients receiving lung cancer surgery. METHODS: We selected 124 patients who received surgery for treating stage I or II lung cancer and divided into experimental group and control group. The experimental group received comprehensive psychological intervention while the control group was given conventional nursing intervention. Pain of patients in two groups was evaluated by visual analog scale (VAS). Before and after intervention, CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+) and free cortisol level in serum were measured. Moreover, QLQ-C30, a life quality measurement scale developed by European Organization for Research and Treatment of Cancer (EORTC) was used. RESULTS: Compared to control group, VAS of patients in experimental group remarkably decreased before anesthesia, 6 hour, 12 hour 24 hour and 48 hour after surgery (P<0.05), and moreover, OLQ-C30 score and various factor scores (except physical symptoms) in experimental group were much higher (P<0.05). No statistical significant difference was found in immune index between two groups before intervention (P>0.05). Differences of CD3(+) and CD4(+) before and after intervention were both statistically significant (P<0.05), so did free cortisol level (P<0.05). CONCLUSION: Comprehensive psychological intervention can effectively relieve pain, improve immune functions and enhance quality of life for patients suffering from lung cancer surgery.

[Application of self-stabilizing zero-profile three-dimensional printed artificial vertebral bodies for treatment of cervical spondylotic myelopathy].

Objective: To evaluate the safety and effectiveness of applying self-stabilizing zero-profile three-dimensional (3D) printed artificial vertebral bodies in anterior cervical corpectomy and fusion (ACCF) for cervical spondylotic myelopathy. Methods: A retrospective analysis was conducted on 37 patients diagnosed with cervical spondylotic myelopathy who underwent single-level ACCF using either self-stabilizing zero-profile 3D-printed artificial vertebral bodies ( n=15, treatment group) or conventional 3D-printed artificial vertebral bodies with titanium plates ( n=22, control group) between January 2022 and February 2023. There was no significant difference in age, gender, lesion segment, disease duration, and preoperative Japanese Orthopedic Association (JOA) score between the two groups ( P>0.05). Operation time, intraoperative bleeding volume, hospitalization costs, JOA score and improvement rate, incidence of postoperative prosthesis subsidence, and interbody fusion were recorded and compared between the two groups. Results: Compared with the control group, the treatment group had significantly shorter operation time and lower hospitalization costs ( P<0.05); there was no significant difference in intraoperative bleeding volume between the two groups ( P>0.05). All patients were followed up, with a follow-up period of 6-21 months in the treatment group (mean, 13.7 months) and 6-19 months in the control group (mean, 12.7 months). No dysphagia occurred in the treatment group, while 5 cases occurred in the control group, with a significant difference in the incidence of dysphagia between the two groups ( P<0.05). At 12 months after operation, both groups showed improvement in JOA scores compared to preoperative scores, with significant differences ( P<0.05); however, there was no significant difference in the JOA scores and improvement rate between the two groups ( P>0.05). Radiographic examinations showed the interbody fusion in both groups, and the difference in the time of interbody fusion was not significant ( P>0.05). At last follow-up, 2 cases in the treatment group and 3 cases in the control group experienced prosthesis subsidence, with no significant difference in the incidence of prosthesis subsidence ( P>0.05). There was no implant displacement or plate-screw fracture during follow-up. Conclusion: The use of self-stabilizing zero-profile 3D-printed artificial vertebral bodies in the treatment of cervical spondylotic myelopathy not only achieves similar effectiveness to 3D-printed artificial vertebral bodies, but also reduces operation time and the incidence of postoperative dysphagia.

Gut Microbiota, Human Blood Metabolites, and Esophageal Cancer: A Mendelian Randomization Study.

BACKGROUND: Unbalances in the gut microbiota have been proposed as a possible cause of esophageal cancer (ESCA), yet the exact causal relationship remains unclear. PURPOSE: To investigate the potential causal relationship between the gut microbiota and ESCA with Mendelian randomization (MR) analysis. METHODS: Genome-wide association studies (GWASs) of 207 gut microbial taxa (5 phyla, 10 classes, 13 orders, 26 families, 48 genera, and 105 species) and 205 gut microbiota metabolic pathways conducted by the Dutch Microbiome Project (DMP) and a FinnGen cohort GWAS of esophageal cancer specified the summary statistics. To investigate the possibility of a mediation effect between the gut microbiota and ESCA, mediation MR analyses were performed for 1091 blood metabolites and 309 metabolite ratios. RESULTS: MR analysis indicated that the relative abundance of 10 gut microbial taxa was associated with ESCA but all the 12 gut microbiota metabolic pathways with ESCA indicated no statistically significant association existing. Two blood metabolites and a metabolite ratio were discovered to be mediating factors in the pathway from gut microbiota to ESCA. CONCLUSION: This research indicated the potential mediating effects of blood metabolites and offered genetic evidence in favor of a causal correlation between gut microbiota and ESCA.

ANKFY1 bridges ATG2A-mediated lipid transfer from endosomes to phagophores.

Macroautophagy is a process that cells engulf cytosolic materials by autophagosomes and deliver them to lysosomes for degradation. The biogenesis of autophagosomes requires ATG2 as a lipid transfer protein to transport lipids from existing membranes to phagophores. It is generally believed that endoplasmic reticulum is the main source for lipid supply of the forming autophagosomes; whether ATG2 can transfer lipids from other organelles to phagophores remains elusive. In this study, we identified a new ATG2A-binding protein, ANKFY1. Depletion of this endosome-localized protein led to the impaired autophagosome growth and the reduced autophagy flux, which largely phenocopied ATG2A/B depletion. A pool of ANKFY1 co-localized with ATG2A between endosomes and phagophores and depletion of UVRAG, ANKFY1 or ATG2A/B led to reduction of PI3P distribution on phagophores. Purified recombinant ANKFY1 bound to PI3P on membrane through its FYVE domain and enhanced ATG2A-mediated lipid transfer between PI3P-containing liposomes. Therefore, we propose that ANKFY1 recruits ATG2A to PI3P-enriched endosomes and promotes ATG2A-mediated lipid transfer from endosomes to phagophores. This finding implicates a new lipid source for ATG2A-mediated phagophore expansion, where endosomes donate PI3P and other lipids to phagophores via lipid transfer.

Supercoiled DNA percentage: A key in-process control of linear DNA template for mRNA drug substance manufacturing.

The development of messenger RNA (mRNA) vaccines and therapeutics necessitates the production of high-quality in vitro-transcribed mRNA drug substance with specific critical quality attributes (CQAs), which are closely tied to the uniformity of linear DNA template. The supercoiled plasmid DNA is the precursor to the linear DNA template, and the supercoiled DNA percentage is commonly regarded as a key in-process control (IPC) during the manufacturing of linear DNA template. In this study, we investigate the influence of supercoiled DNA percentage on key mRNA CQAs, including purity, capping efficiency, double-stranded RNA (dsRNA), and distribution of poly(A) tail. Our findings reveal a significant impact of supercoiled DNA percentage on mRNA purity and in vitro transcription yield. Notably, we observe that the impact on mRNA purity can be mitigated through oligo-dT chromatography, alleviating the tight range of DNA supercoiled percentage to some extent. Overall, this study provides valuable insights into IPC strategies for DNA template chemistry, manufacturing, and controls (CMC) and process development for mRNA drug substance.

Identification of long non-coding RNA and circular RNA associated networks in cellular stress responses.

Mammalian cells employ various adaptive responses to cope with multiple stresses to maintain homeostasis. Functional roles of non-coding RNAs (ncRNAs) in response to cellular stresses have been proposed, and systematical investigations about the crosstalk among distinct types of RNAs are required. Here, we challenged HeLa cells with thapsigargin (TG) and glucose deprivation (GD) treatments to induce endoplasmic reticulum (ER) and metabolic stresses, respectively. Ribosomal RNA (rRNA)-depleted RNA sequencing (RNA-seq) was then performed. Characterization of the RNA-seq data revealed a series of differentially expressed long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) with parallel changes responsive to both stimuli. We further constructed the lncRNA/circRNA-mRNA co-expressing network, competing endogenous RNA (ceRNA) network in the lncRNA/circRNA-miRNA-mRNA axis, and lncRNA/circRNA-RNA binding protein (RBP) interactome map. These networks indicated the potential cis and/or trans regulatory roles of lncRNAs and circRNAs. Moreover, Gene Ontology analysis demonstrated that these identified ncRNAs were associated with several essential biological processes known to be related to cellular stress responses. In conclusion, we systematically established functional regulatory networks of lncRNA/circRNA-mRNA, lncRNA/circRNA-miRNA-mRNA and lncRNA/circRNA-RBP to perceive the potential interactions and biological processes during cellular stresses. These results provided insights in ncRNA regulatory networks of stress responses and the basis for further identification of pivotal factors involved in cellular stress responses.

Realizing Photocatalytic Overall Water Splitting by Modulating the Thickness-Induced Reaction Energy Barrier of Fluorenone-Based Covalent Organic Frameworks.

Direct photocatalytic hydrogen and oxygen evolution from water splitting is an attractive approach for producing chemical fuels. In this work, a novel fluorenone-based covalent organic framework (COF-SCAU-2) is successfully exfoliated into ultrathin three-layer nanosheets (UCOF-SCAU-2) for photocatalytic overall water splitting (OWS) under visible light. The ultrathin structures of UCOF-SCAU-2 greatly enhance carrier separation, utilization efficiency, and the exposure of active surface sites. Surprisingly, UCOF-SCAU-2 exhibits efficient photocatalytic OWS performance, with hydrogen and oxygen evolution rates reaching 0.046 and 0.021 mmol h-1 g-1 , respectively, under visible-light irradiation, whereas bulk COF-SCAU-2 shows no activity for photocatalytic OWS. Charge-carrier kinetic analysis and DFT calculations confirm that reducing the thickness of the COF nanosheets increases the number of accessible active sites, reduces the distance for charge migration, prolongs the lifetimes of photogenerated carriers, and decreases the Gibbs free energy of the rate-limiting step compared to nonexfoliated COFs. This work offers new insights into the effect of the layer thickness of COFs on photocatalytic OWS.

Using Piezosurgery in Anterior Cervical Discectomy and Fusion to Treat Complex Cervical Spondylotic Myelopathy Is Safe and Effective.

Objective: To investigate the safety and efficacy of piezosurgery in anterior cervical discectomy and fusion (ACDF) for cervical spondylotic myelopathy (CSM). Methods: 47 patients with complex CSM (cCSM) underwent ACDF surgery from 2014 to 2017. Among these patients, 26 underwent ACDF using piezosurgery (group A) and 21 underwent ACDF by using traditional tools such as high-speed air drill, bone curette, and Kerrison bone punch (group B). Average surgical time, intraoperative blood loss, surgical complications, preoperative and postoperative Japanese Orthopaedic Association (JOA) scores, and improvement rate were measured. Results: Average surgical time and intraoperative blood loss were significantly lower in group A than those in group B (P < 0.01). The incidences of surgical complications were 3.8% and 23.8% in the A and B groups (P < 0.05), respectively. There were no significant differences in JOA scores and improvement rates between data collection periods at preoperative, 3-day postoperative, and 1-year postoperative follow-ups (P > 0.05). Conclusion: For treating cCSM, both the piezosurgery and traditional tools led to significant neurological improvement. However, the piezosurgery was superior to the traditional tools in terms of surgical time, blood loss, and complication rate. Hence, piezosurgery was a safe and effective adjunct for ACDF treating cCSM.

Lysine crotonylation regulates leucine-deprivation-induced autophagy by a 14-3-3ε-PPM1B axis.

Lysine crotonylation as a protein post-translational modification regulates diverse cellular processes and functions. However, the role of crotonylation in nutrient signaling pathways remains unclear. Here, we find a positive correlation between global crotonylation levels and leucine-deprivation-induced autophagy. Crotonylome profiling identifies many crotonylated proteins regulated by leucine deprivation. Bioinformatics analysis dominates 14-3-3 proteins in leucine-mediated crotonylome. Expression of 14-3-3ε crotonylation-deficient mutant significantly inhibits leucine-deprivation-induced autophagy. Molecular dynamics analysis shows that crotonylation increases molecular instability and disrupts the 14-3-3ε amphipathic pocket through which 14-3-3ε interacts with binding partners. Leucine-deprivation-induced 14-3-3ε crotonylation leads to the release of protein phosphatase 1B (PPM1B) from 14-3-3ε interaction. Active PPM1B dephosphorylates ULK1 and subsequently initiates autophagy. We further find that 14-3-3ε crotonylation is regulated by HDAC7. Taken together, our findings demonstrate that the 14-3-3ε-PPM1B axis regulated by crotonylation may play a vital role in leucine-deprivation-induced autophagy.

KAT7-mediated CANX (calnexin) crotonylation regulates leucine-stimulated MTORC1 activity.

Amino acids play crucial roles in the MTOR (mechanistic target of rapamycin kinase) complex 1 (MTORC1) pathway. However, the underlying mechanisms are not fully understood. Here, we establish a cell-free system to mimic the activation of MTORC1, by which we identify CANX (calnexin) as an essential regulator for leucine-stimulated MTORC1 pathway. CANX translocates to lysosomes after leucine deprivation, and its loss of function renders either the MTORC1 activity or the lysosomal translocation of MTOR insensitive to leucine deprivation. We further find that CANX binds to LAMP2 (lysosomal associated membrane protein 2), and LAMP2 is required for leucine deprivation-induced CANX interaction with the Ragulator to inhibit Ragulator activity toward RRAG GTPases. Moreover, leucine deprivation promotes the lysine (K) 525 crotonylation of CANX, which is another essential condition for the lysosomal translocation of CANX. Finally, we find that KAT7 (lysine acetyltransferase 7) mediates the K525 crotonylation of CANX. Loss of KAT7 renders the MTORC1 insensitivity to leucine deprivation. Our findings provide new insights for the regulatory mechanism of the leucine-stimulated MTORC1 pathway.Abbreviations: CALR: calreticulin; CANX: calnexin; CLF: crude lysosome fraction; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; ER: endoplasmic reticulum; GST: glutathione S-transferase; HA: hemagglutinin; HEK293T: human embryonic kidney-293T; KAT7: lysine acetyltransferase 7; Kcr; lysine crotonylation; KO: knockout; LAMP2: lysosomal associated membrane protein 2; LAMTOR/Ragulator: late endosomal/lysosomal adaptor: MAPK and MTOR activator; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MTOR: mechanistic target of rapamycin kinase; PDI: protein disulfide isomerase; PTM: post-translational modification; RPS6KB1/p70S6 kinase 1: ribosomal protein S6 kinase B1; RPTOR: regulatory associated protein of MTOR complex 1; SESN2: sestrin 2; TMEM192: transmembrane protein 192; ULK1: unc-51 like autophagy activating kinase 1.

Exportin 4 depletion leads to nuclear accumulation of a subset of circular RNAs.

Numerous RNAs are exported from the nucleus, abnormalities of which lead to cellular complications and diseases. How thousands of circular RNAs (circRNAs) are exported from the nucleus remains elusive. Here, we provide lines of evidence to demonstrate a link between the conserved Exportin 4 (XPO4) and nuclear export of a subset of circRNAs in metazoans. Exonic circRNAs (ecircRNAs) with higher expression levels, larger length, and lower GC content are more sensitive to XPO4 deficiency. Cellular insufficiency of XPO4 leads to nuclear circRNA accumulation, circRNA:DNA (ciR-loop) formation, linear RNA:DNA (liR-loop) buildup, and DNA damage. DDX39 known to modulate circRNA export can resolve ciR-loop, and splicing factors involved in the biogenesis of circRNAs can also affect the levels of ciR-loop. Testis and brain are two organs with high abundance of circRNAs, and insufficient XPO4 levels are detrimental, as Xpo4 heterozygous mice display male infertility and neural phenotypes. Increased levels of ciR-loop, R-loop, and DNA damage along with decreased cell numbers are observed in testis and hippocampus of Xpo4 heterozygotes. This study sheds light on the understandings of mechanism of circRNA export and reveals the significance of efficient nuclear export of circRNAs in cellular physiology.

PFKP facilitates ATG4B phosphorylation during amino acid deprivation-induced autophagy.

ATG4B facilitates autophagy by promoting autophagosome maturation through the reversible lipidation and delipidation of LC3. Recent reports have shown that phosphorylation of ATG4B regulates its activity and LC3 processing, leading to modulate autophagy activity. However, the mechanism about how ATG4B phosphorylation is involved in amino acid deprivation-induced autophagy is unclear. Here, we combined the tandem affinity purification with mass spectrometry (MS) and identified the ATG4B-interacting proteins including its well-known partner gamma-aminobutyric acid receptor-associated protein (GABARAP, a homolog of LC3) and phosphofructokinase 1 platelet isoform (PFKP). Further immunoprecipitation assays showed that amino acid deprivation strengthened the interaction between ATG4B and PFKP. By genetic depletion of PFKP using CRISPR/Cas9, we uncovered that PFKP loss reduced the degradation of LC3-II and p62 due to a partial block in autophagic flux. Furthermore, MS analysis of Flag-tagged ATG4B immunoprecipitates identified phosphorylation of ATG4B serine 34 residue (S34) and PFKP serine 386 residue (S386) under amino acid deprivation condition. In vitro kinase assay validated that PFKP functioning as a protein kinase phosphorylated ATG4B at S34. This phosphorylation could enhance ATG4B activity and p62 degradation. In addition, PFKP S386 phosphorylation was important to ATG4B S34 phosphorylation and autophagy in HEK293T cells. In brief, our findings describe that PFKP, a rate-limiting enzyme in the glycolytic pathway, functions as a protein kinase for ATG4B to regulate ATG4B activity and autophagy under amino acid deprivation condition.

Lysobacter may drive the hormetic effects of Pb on soil alkaline phosphatase.

It has become increasingly recognized that hormesis phenomena exist in soil ecosystem, but the research on the hormetic responses of soil enzymes are still limited. This study was conducted to investigate the hormetic effects of lead (Pb) on the activity of soil alkaline phosphatase (ALP) and the associated microbial groups. Soils were treated by adding Pb (NO3)2 solution with 0, 10, 100, 500, 1000, 2000, 4000, and 5000 mg/kg of Pb, respectively. A moist heat sterilization method (121 °C × 30 min) was used to discriminate the microbial effect on soil ALP hormesis from other factors. The bacterial community composition and abundance in the control (CK) and Pb-treated soils were detected by the high-throughput sequencing technique. The ALP activity at doses of 500-1000 mg/kg of Pb was significantly higher than that of CK (0 mg/kg of Pb), showing a typical inverted U-shaped dose response with the stimulation magnitude of 9.8-10.3% within 48 h of incubation. In addition, ALP activity decreased by 80% on average after soil sterilization. Analysis of bacterial community composition indicated that the relative abundance of Lysobacter at 1000 mg Pb/kg was higher than that of CK at genus level, with the increase of 69.82%. The highly significant correlation between soil ALP activities and relative abundance of Lysobacter indicated that this bacterial genus could possibly contribute to the hormetic responses of soil ALP to added doses of Pb in soils.