Mass-Spectrometry-Based Assay at Single-Base Resolution for Simultaneously Detecting m6A and m6Am in RNA.

The methylation modifications of adenosine, especially N6-methyladenosine (m6A) and N6, 2'-odimethyladenosine (m6Am), play vital roles in various biological, physiological, and pathological processes. However, current methods for detecting these modifications at single-base resolution have limitations. Mass spectrometry (MS), a highly accurate and sensitive technique, can be utilized to differentiate between m6A and m6Am by analyzing the molecular weight differences in their fragments during tandem MS analysis. In this study, we present an MS-based method that allows for the simultaneous determination of m6A and m6Am sites in targeted RNA fragments at single-nucleotide resolution. The approach involves the utilization of tandem MS in conjunction with targeted RNA enrichment and enzymatic digestion, eliminating the need for PCR amplification. By employing this strategy, we can accurately identify m6A and m6Am sites in targeted RNA fragments with high confidence. To evaluate the effectiveness of our method, we applied it to detect m6A and m6Am sites in cell and tissue samples. Furthermore, we verified the accuracy of our approach by performing CRISPR/Cas9-mediated knockout of the corresponding methyltransferases. Overall, our MS-based method offers a reliable and precise means for the simultaneous detection of m6A and m6Am modifications in targeted RNA fragments, providing valuable insights into the functional characterization of these modifications in various biological contexts.

Boosting Electrochemiluminescence Performance of a Dual-Active Site Iron Single-Atom Catalyst-Based Luminol-Dissolved Oxygen System via Plasmon-Induced Hot Holes.

Due to the commonly low content of biomarkers in diseases, increasing the sensitivity of electrochemiluminescence (ECL) systems is of great significance for in vitro ECL diagnosis and biodetection. Although dissolved O2 (DO) has recently been considered superior to H2O2 as a coreactant in the most widely used luminol ECL systems owing to its improved stability and less biotoxicity, it still has unsatisfactory ECL performance because of its ultralow reactivity. In this study, an effective plasmonic luminol-DO ECL system has been developed by complexing luminol-capped Ag nanoparticles (AgNPs) with plasma-treated Fe single-atom catalysts (Fe-SACs) embedded in graphitic carbon nitride (g-CN) (pFe-g-CN). Under optimal conditions, the performance of the resulting ECL system could be markedly increased up to 1300-fold compared to the traditional luminol-DO system. Further investigations revealed that duple binding sites of pFe-g-CN and plasmonically induced hot holes that disseminated from AgNPs to g-CN surfaces lead to facilitate significantly the luminous reaction process of the system. The proposed luminol-DO ECL system was further employed for the stable and ultrasensitive detection of prostate-specific antigen in a wide linear range of 1.0 fg/mL to 1 µg/mL, with a pretty low limit of detection of 0.183 fg/mL.

Research on proactive defense and dynamic repair of complex networks considering cascading effects.

Cascading effects can result in the nonlinear propagation of failures in complex networks, ultimately leading to network collapse. Research on the fault propagation principles, defense strategies, and repair strategies can help mitigate the effects of cascading failures. Especially, proactive defense and dynamic repair are flexible and effective methods to ensure network security. Most studies on the cascade of complex networks are based on the unprocessed initial information of the network. However, marginal nodes are a type of node that cloaks the initial information of the network. In this study, we rank the importance of nodes according to the intensity of network energy confusion after the removal of this node, clarify the meaning of marginal nodes and proposed two methods to screen marginal nodes. The results indicated that the proactive removal of marginal nodes can effectively reduce the effect of cascading failures without causing any negative disturbance to the energy flow of the network. In addition, network repair according to the proposed strategy can minimize the cascade effect in the repair process and improve repair efficiency.

Effects of Varicocele Surgical Repair on Serum Hormone and Inhibin B Levels for Patients With Varicocele: A Systematic Review and Meta-Analysis.

Varicocele surgical repair can improve the function of the testis for patients with varicocele. We carried out a systematic review and meta-analysis to assess the effects of varicocele surgical repair on serum hormones and inhibin B levels in patients with varicocele. A literature search was performed in August 2022, and no language or geographic region restrictions were applied. The search included the following databases: PubMed, Embase, and Medline. A literature review was performed to identify all published clinical trials assessing serum hormone and inhibin B levels before and after varicocele surgical repair. The reference lists of retrieved studies were also investigated. A systematic review and meta-analysis were conducted. Eight articles were selected from 162 articles, including 452 patients. The combined analysis showed that after surgical treatment, mean serum testosterone, inhibin B, and sperm concentration levels increased compared with preoperative levels (p < .05). After surgical treatment, mean serum follicle-stimulating hormon (FSH), and Luteinizing hormone (LH) levels decreased compared with preoperative levels (p < .05). This meta-analysis demonstrates that varicocele surgical repair can improve testicular function, increase serum testosterone, and inhibin B levels and decrease serum FSH and LH levels in patients with varicocele. This might be related to the improvement of infertility. A large-scale multicenter randomized controlled study is needed for further confirmation.

Deamination-triggered exponential signal amplification for chemiluminescent detection of cytosine deaminase at the single-cell level.

We construct a sensitive chemiluminescent biosensor for sensitive detection of cytosine deaminase APOBEC3A based on deamination-triggered exponential signal amplification. This biosensor displays good specificity and high sensitivity, and it can screen APOBEC3A inhibitors and measure endogenous APOBEC3A at the single-cell level, with prospective applications in disease diagnostics and therapy.

The efficacy of CO2 laser in the treatment of genitourinary syndrome of menopause: a systematic review and meta-analysis of randomized controlled trials.

CO2 laser has been proposed as a treatment strategy for genitourinary syndrome of menopause (GSM). In order to assess its efficacy for treating GSM, we conducted a systematic review and meta-analysis. To identify the current state of randomized controlled trials on CO2 laser therapy for GSM, a literature review was conducted. We systematically searched the following databases: PUBMED, EMBASE and the Cochrane Controlled Trials Register. In addition, a review of the references in the retrieved studies was carried out. Of 562 identified studies, 9 were eligible and were included in our analysis, involving 523 patients in total. Based on our analysis, CO2 laser has no statistical difference compared with estrogen in VHI (p = 0.87), FSFI total score (p = 0.19), FSFI-Arousal (p = 0.11), FSFI-Desire (p = 0.72), FSFI-Orgasm (p = 0.45) and FSFI-Satisfaction (p = 0.08). The meta-analysis also showed that CO2 laser significantly improved FSFI-Lubrication scores compared with estrogen therapy (p = 0.0004). Furthermore, compared with the sham group, CO2 laser group had statistically improved VHI scores (p = 0.003) and FSFI scores (p < 0.00001). CO2 laser therapy may be an effective alternative to estrogen therapy for GSM both in cases where estrogen is not applicable because of comorbidities and in cases in which women do not desire to take estrogen.

Machine Learning-Based Label-Free SERS Profiling of Exosomes for Accurate Fuzzy Diagnosis of Cancer and Dynamic Monitoring of Drug Therapeutic Processes.

Exosomes are a class of extracellular vesicles secreted by cells, which can be used as promising noninvasive biomarkers for the early diagnosis and treatment of diseases, especially cancer. However, due to the heterogeneity of exosomes, it remains a grand challenge to distinguish accurately and reliably exosomes from clinical samples. Herein, we achieve accurate fuzzy discrimination of exosomes from human serum samples for accurate diagnosis of breast cancer and cervical cancer through machine learning-based label-free surface-enhanced Raman spectroscopy (SERS), by using "hot spot" rich 3D plasmonic AuNPs nanomembranes as substrates. Due to the existence of some weak distinguishable SERS fingerprint signals and the high sensitivity of the method, the machine learning-based SERS analysis can precisely identify three (normal and cancerous) cell lines, two of which are different types of cancer cells, without specific labeling of biomarkers. The prediction accuracy based on the machine learning algorithm was up to 91.1% for the discrimination of different cell lines (H8, HeLa, and MCF-7 cell)-derived exosomes. Our model trained with SERS spectra of cell-derived exosomes could reach 93.3% prediction accuracy for clinical samples. Furthermore, the action mechanism of the chemotherapeutic process of MCF-7 cells can be revealed by dynamic monitoring of SERS profiling of the exosomes secreted. The method would be useful for noninvasive and accurate diagnosis and postoperative assessment of cancer or other diseases in the future.

The parABSm system is involved in megaplasmid partitioning and genome integrity maintenance in Thermus thermophilus.

The characteristics of the parABS system in polyploid bacteria are barely understood. We initially analyzed the physiological functions and mechanisms of the megaplasmid parABSm system in the thermophilic polyploid bacterium Thermus thermophilus. Deletion of parABm was possible only when a plasmid-born copy of parABm was provided, indicating that these genes are conditionally essential. The cell morphology of the parABm deletion mutant (ΔparABm) was changed to some extent, and in certain extra-large or twisted cells, the nucleoids were dispersed and damaged. Compared with that of the wild type, the frequency of anucleate cells was significantly increased. Genome content analyses showed that ΔparABm had lost ∼160 kb of megaplasmid and ∼23 kb of chromosomal sequences, respectively. Genome fluorescent tagging and PFGE experiments demonstrated that the truncated megaplasmid was frequently interlinked and could not be segregated correctly; thus, certain daughter cells eventually lost the entire megaplasmid and became twisted or enlarged with damaged nucleoids. Further, we found that when the megaplasmid was lost in these cells, the toxins encoded by the megaplasmid toxin-antitoxin (TA) systems (VapBC64_65 and VapBC142_143) would exert detrimental effects, such as to fragment DNA. Thus, parABSm might ensure the existence of these TA systems, thereby preventing genomic degradation. Together, our results suggested that in T. thermophilus, the megaplasmid-encoded parABS system plays an essential role in the megaplasmid partitioning process; also it is an important determination factor for the genome integrity maintenance.

Glass Capillary-Based Nanopores for Single Molecule/Single Cell Detection.

A glass capillary-based nanopore (G-nanopore), due to its tapered tip, easy tunability in orifice size, and especially its flexible surface modifications that can be tailored to effectively capture and enhance the ionic current signal of single entities (single molecules, single cells, and single particles), offers a powerful and nanoconfined sensing platform for diverse biological measurements of single cells and single molecules. Compared with other artificial two-dimensional solid-state nanopores, its conical tip and high spatial and temporal resolution characteristics facilitate noninvasive single molecule and selected area (subcellular) single cell detections (e.g., DNA mutations, highly expressed proteins, and small molecule markers that reflect the change characteristics of the tumor), as a small G-nanopore (≤100 nm) does negligible damage to cell functions and cell membrane integrity when inserted through the cell membrane. In this brief review, we summarize the preparation of G-nanopores and discuss the advantages of them as solid-state sensing platforms for single molecule and single cell detection applications as well as for cancer diagnosis and treatment applications. We also describe the current bottlenecks that limit the widespread use of G-nanopores in clinical applications and provide an outlook on future developments. The brief review will provide the reader with a quick survey of this field and facilitate the rapid development of a G-nanopore sensing platform for future tumor diagnosis and personalized medicine based on single-molecule/single-cell bioassay.

Solid-Phase Pyrolysis Synthesis of Highly Fluorescent Nitrogen/Sulfur Codoped Graphene Quantum Dots for Selective and Sensitive Diversity Detection of Cr(VI).

In this study, a simple one-step solid-phase pyrolysis synthesis procedure was employed to prepare N and S codoped highly fluorescent graphene quantum dots (N/S-GQDs). The as-synthesized quantum dot showed λexcitation-dependent blue fluorescence (FL) emission with a relative quantum yield of about 22% and displayed good biocompatibility, high water dispersibility, and excellent stability under extreme conditions (i.e., ionic strength, pH, and temperature). The potential applicability of the as-synthesized quantum dot was tested by employing solution- and paper-based FL detection modes for Cr(VI) detection. The proposed solution- and paper-based FL sensors showed lower limit of detection (LOD) values of 0.01 and 0.4 µM, respectively. The as-constructed paper- and solution-based FL sensors proved the feasibility of sensitive, cost-effective, and on-site detection of Cr(VI).

Gold nanoparticle-based signal amplified electrochemiluminescence for biosensing applications.

Since the content levels of biomarkers at the early stage of many diseases are generally lower than the detection threshold concentration, achieving ultrasensitive and accurate detection of these biomarkers is still one of the major goals in bio-analysis. To achieve ultrasensitive and reliable bioassay, it requires developing highly sensitive biosensors. Among all kinds of biosensors, electrogenerated chemiluminescence (ECL) based biosensors have attracted enormous attention due to their excellent properties. In order to improve the performance of ECL biosensors, gold nanoparticles (Au NPs) have been widely utilized as signal amplification tags. The introduction of Au NPs could dramatically enhance the performance of the constructed ECL biosensors via diverse ways such as electrode modification material, efficient energy acceptor in ECL resonant energy transfer (ECL-RET), reaction catalyst, surface plasmon resonance (SPR) enhancer, and as nanocarrier. Herein, we summarize recent developments and progress of ECL biosensors based on Au NPs signal amplification strategies. We will cover ECL applications of Au NPs as a signal amplification tag in the detection of proteins, metal ions, nucleic acids, small molecules, living cells, exosomes, and cell imaging. Finally, brief summary and future outlooks of this field will be presented.

Plasmon-Boosted Fe, Co Dual Single-Atom Catalysts for Ultrasensitive Luminol-Dissolved O2 Electrochemiluminescence Detection of Prostate-Specific Antigen.

Improving the sensitivity of electrochemiluminescence (ECL) systems is highly desired for in vitro ECL diagnosis and bio-detections due to the often-low content of biomarkers in diseases. And dissolved O2 (DO) as a co-reactant is considered superior to H2O2 in the most commonly used luminol ECL systems due to better stability and low biotoxicity, but it still suffers from low ECL performance due to the low reactivity of DO. In this study, an efficient luminol-DO ECL system was developed through the complexing of Fe, Co dual single-atom catalysts (D-SACs) supported by N-doped graphene with the luminol-capped Ag nanoparticles (AgNPs). Benefiting from the electronic interaction between Fe and Co metal sites in the relevant D-SACs and plasmon enhancement of AgNPs, the performance of the corresponding ECL system could be significantly boosted up to ≈677-fold under optimal testing conditions, comparable to the classic luminol-O2 system. Furthermore, the developed luminol-DO ECL system was successfully applied for the stable ultrasensitive detection of prostate-specific antigen (PSA) in a wide linear range of 1 fg/mL to 1 µg/mL, with a low limit of detection (0.98 fg/mL).

Associations Between a Polymorphism in the Rat 5-HT1A Receptor Gene Promoter Region (rs198585630) and Cognitive Alterations Induced by Microwave Exposure.

The nervous system is a sensitive target of electromagnetic radiation (EMR). Chronic microwave exposure can induce cognitive deficits, and 5-HT system is involved in this effect. Genetic polymorphisms lead to individual differences. In this study, we evaluated whether the single-nucleotide polymorphism (SNP) rs198585630 of 5-HT1A receptor is associated with cognitive alterations in rats after microwave exposure with a frequency of 2.856 GHz and an average power density of 30 mW/cm2. Rats were exposed to microwaves for 6 min three times a week for up to 6 weeks. PC12 cells and 293T cells were exposed to microwaves for 5 min up to 3 times at 2 intervals of 5 min. Transcriptional activity of 5-HT1A receptor promoter containing rs198585630 C/T allele was determined in vitro. Electroencephalograms (EEGs), spatial learning and memory, and mRNA and protein expression of 5-HT1A receptor were evaluated in vivo. We demonstrated that transcriptional activity of 5-HT1A receptor promoter containing rs198585630 C allele was higher than that of 5-HT1A receptor promoter containing T allele. The transcriptional activity of 5-HT1A receptor promoter was stimulated by 30 mW/cm2 microwave exposure, and rs198585630 C allele was more sensitive to microwave exposure, as it showed stronger transcriptional activation. Rats carrying rs198585630 C allele exhibited increased mRNA and protein expression of 5-HT1A receptor and were more susceptible to 30 mW/cm2 microwave exposure, showing cognitive deficits and inhibition of brain electrical activity. These findings suggest SNP rs198585630 of the 5-HT1A receptor is an important target for further research exploring the mechanisms of hypersensitivity to microwave exposure.

Photoactivated Bacteriorhodopsin/SiNx Nanopore-Based Biological Nanofluidic Generator with Single-Protein Sensitivity.

Nanofluidics is an emerging hot field that explores the unusual behaviors of ions/molecules transporting through nanoscale channels, which possesses a broad application prospect. However, in situ probing bioactivity of functional proteins on a single-molecule level by a nanofluidic device has not been reported, and it is still a big challenge in the field. Herein, we reported a biological nanofluidic device with a single-protein sensitivity, based on natural proton-pumping protein, bacteriorhodopsin (bR), and a single SiNx nanopore. Nanofluidic single-molecule probing of bR proton-pumping activity and its light response were achieved under applied voltage of 0 V, by biologically self-powered steady-state ionic current nanopore sensing. Green-light irradiation of the device led to the monitoring of a steady-state proton current of ∼3.51 pA/per bR trimer, corresponding to charge density of 815 µC/cm2 generated by each bR monomer, which far exceeded the previously reported value of 1.4 µC/cm2. This finding and method would promote the development of artificial biological and hybrid nanofluidic devices in biosensing and energy conversion applications.

Ultrahigh Elastic Polymer Electrolytes for Solid-State Lithium Batteries with Robust Interfaces.

Solid polymer electrolytes (SPEs) are promising for solid-state lithium batteries, but their practical application is significantly impeded by their low ionic conductivity and poor compatibility. Here, we report an ultrahigh elastic SPE based on cross-linked polyurethane (PU), succinonitrile (SN), and lithium bistrifluoromethanesulfonimide (LiTFSI). The resulting electrolyte (PU-SN-LiTFSI) exhibits an ionic conductivity of 2.86 × 10-4 S cm-1, a tensile strength of 3.8 MPa, and a breaking elongation exceeding 3000% at room temperature. A solid-state lithium battery using the electrolyte exhibits a high specific capacity of 150 mAh g-1 at 0.2C and a long cycling life of up to 700 cycles at 0.5C at room temperature, showing one of the best performances among its counterparts. The excellent performances are attributed to the fact that its ultrahigh elasticity, good ionic conductivity, tensile strength, and electrochemical stability contribute to robust electrode/electrolyte interfaces, thus greatly decreasing the charge-transfer resistance in charge/discharge processes. Our investigations provide a novel strategy to address the intrinsic interfacial issue of solid-state batteries.

Surface-Bonding-Enhanced Self-Co-Reactant Electrogenerated Chemiluminescence for Sensitive and Selective Detection of Thioglycolic Acid in Cosmetics.

Thioglycolic acid (TGA) is an organic compound widely used in cosmetics that cause a variety of health problems when overexposed to it. So far many attempts have been made to develop methods for TGA detection, but most of them need sophisticated instrumentations and are a little bit complicated. Therefore, a simple, cheap and sensitive detection method of TGA is highly desired. Herein, we demonstrated for the first time an Au-S bonding amplified, highly sensitive electrochemiluminescence (ECL) sensing method for TGA detection using tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)3 2+ ) as a luminophore and TGA as a self-co-reactant, via an anodic reaction at the Au electrode surface. Due to different molecular coordination environments of the TGA at the electrode surface, the ECL signal intensity of the developed ECL system gives much higher ECL signal in borate buffer than phosphate buffer of the same pH. Under the optimized experimental conditions, the ECL intensity has a direct relationship with the concentration of TGA in the range of 0.03 µM to 300 µM and a limit of detection of 0.013 µM (3σ/m). The reported ECL system has further been applied for the detection of TGA in cosmetics with acceptable recoveries.

Plasmon-Enhanced Nitrogen Vacancy-Rich Carbon Nitride Electrochemiluminescence Aptasensor for Highly Sensitive Detection of miRNA.

The development of biosensors for biologically important substances with ultralow content such as microRNA is of great significance. Herein, a novel surface plasmon-enhanced electrogenerated chemiluminescence-based aptasensor was developed for ultrasensitive sensing of microRNA by using nitrogen vacancy-rich carbon nitride nanosheets as effective luminophores and gold nanoparticles as plasmonic sources. The introduction of nitrogen vacancies improved the electrochemiluminescence behavior due to improved conductance and electrogenerated chemiluminescence activity. The introduction of plasmonic gold nanoparticles increased the electrochemiluminescence signal intensity by more than eightfold. The developed surface plasmon-enhanced electrogenerated chemiluminescence aptasensor exhibited good selectivity, ultrasensitivity, excellent stability, and reproducibility for the determination of microRNA-133a, with a dynamic linear range of 1 aM to 100 pM and a limit of detection about 0.87 aM. Moreover, the surface plasmon-enhanced electrogenerated chemiluminescence sensor obtained a good recovery when detecting the content of microRNA in actual serum.

High-efficiency genome editing of an extreme thermophile Thermus thermophilus using endogenous type I and type III CRISPR-Cas systems.

Thermus thermophilus is an attractive species in the bioindustry due to its valuable natural products, abundant thermophilic enzymes, and promising fermentation capacities. However, efficient and versatile genome editing tools are not available for this species. In this study, we developed an efficient genome editing tool for T. thermophilus HB27 based on its endogenous type I-B, I-C, and III-A/B CRISPR-Cas systems. First, we systematically characterized the DNA interference capabilities of the different types of the native CRISPR-Cas systems in T. thermophilus HB27. We found that genomic manipulations such as gene deletion, mutation, and in situ tagging could be easily implemented by a series of genome-editing plasmids carrying an artificial self-targeting mini-CRISPR and a donor DNA responsible for the recombinant recovery. We also compared the genome editing efficiency of different CRISPR-Cas systems and the editing plasmids with donor DNAs of different lengths. Additionally, we developed a reporter gene system for T. thermophilus based on a heat-stable ß-galactosidase gene TTP0042, and constructed an engineered strain with a high production capacity of superoxide dismutases by genome modification.

Effects of heavy metals on bacterial community structures in two lead-zinc tailings situated in northwestern China.

We evaluated the variations of bacterial communities in six heavy metal contaminated soils sampled from Yanzi Bian (YZB) and Shanping Cun (SPC) tailings located in northwestern China. Statistical analysis showed that both the heavy metals and soil chemical properties could affect the structure and diversity of the bacterial communities in the tailing soils. Cd, Cu, Zn, Cr, Pb, pH, SOM (soil organic matters), TP (total phosphorus) and TN (total nitrogen) were the main driving factors of the bacterial community variations. As a consequence, the relative abundances of certain bacterial phyla including Proteobacteria, Chloroflexi, Firmicutes, Nitrospirota and Bacteroidota were significantly increased in the tailing soils. Further, we found that the abundance increasement of these phyla were mainly contributed by certain species, such as s__unclassified_g__Thiobacillus (Proteobacteria), s__unclassified_g__Sulfobacillus (Firmicutes) and Leptospirillum ferriphilum (Nitrospirota). Thus, these species were considered to be strongly heavy metal tolerant. Together, our findings will provide a useful insight for further bioremediations of these contaminated areas.

Plasmon-Boosted Cu-Doped TiO2 Oxygen Vacancy-Rich Luminol Electrochemiluminescence for Highly Sensitive Detection of Alkaline Phosphatase.

In this study, an effective oxygen vacancy (Ov)-involved luminol-dissolved oxygen (O2) electrochemiluminescence (luminol-DO ECL) system was developed and exploited for ECL sensing applications through significant plasmon enhancement of the Ov-involved weak luminol-DO ECL signals by the combined use of Cu-doped TiO2 oxygen vacancy and a Au@SiO2 nanomembrane. The results disclosed that the ECL response of the corresponding system could be synergistically boosted, and the plausible underlying mechanism has been discussed. Furthermore, for the first time, the developed system has been successfully applied for the highly sensitive detection of alkaline phosphatase with a low limit of detection of 0.005 U/L, with an excellent linear range from 0.005 to 10 U/L, as well as good stability and reproducibility.