Epigenetic Age Estimation by Detecting DNA Methylation Status in Buccal Swabs.

The analysis of DNA methylation (DNAm) levels at specific CpG sites represents one of the most promising molecular techniques for estimating an individual's age. To date, a considerable number of studies have reported the development of age prediction models on the basis of DNAm in body fluids, with only a few utilizing buccal swabs. The objective of this study was to identify age-dependent methylation CpG sites in three different genes (HOXC4, TRIM59, and ELOVL2) in buccal swab samples from the Chinese Han population. A total of 461 buccal swabs, with an age range of 0.4-80.8 years, were divided into a training set (n = 325) and a validation set (n = 136). Samples were analyzed by pyrosequencing in order to identify age-related genes with correlation coefficient. A random forest regression model was ultimately proposed, including eight CpGs in three genes, with a mean absolute error (MAE) of 2.119 years. The model performs independent validation set with an MAE of 4.391 years. Our findings illustrate that buccal swabs present a suitable alternative to biological traces for age prediction based on DNAm pattern using pyrosequencing and random forest regression, offering the additional advantage of being collected noninvasively.

Upconversion Phosphor-Driven Photodegradation of Plastics.

Plastic waste poses a profound threat to ecosystems and human health, necessitating novel strategies for effective degradation in nature. Here, we present a novel approach utilizing upconversion phosphors as additives to significantly accelerate plastic photodegradation in nature via enhancing ultraviolet (UV) radiation. Pr-doped Li2CaGeO4 (LCGO:Pr) upconversion phosphors readily converting blue light into deep-UV radiation, dramatically improve photodegradation rates for polyethylene (PE) and polyethylene terephthalate (PET) microplastics. In situ spectroscopic studies show that upconversion fluorescence initiates the photophysical cleavage of C-C and C-O bonds in the backbones of PE and PET, resulting in plastic degradation. Moreover, incorporating LCGO:Pr into polypropylene (PP) sheets realizes markedly enhanced photodamage, with the cracking area increasing by nearly 38-fold under simulated sunlight for 10 days. This underscores the potential of employing this approach for the construction of light-driven destructible polymers. Further optimization and exploration of material compatibility hold promise for developing sustainable photodegradable plastics.

Simultaneous removal of nitrate, copper, carbamazepine, and calcium from micropolluted water by fulvic acid through promotion of denitrification and microbial-induced calcium precipitation: Performance and mechanism.

In this paper, a study of the denitrification strain Cupriavidus sp. W12 was conducted to remove copper (Cu2+), carbamazepine (CBZ), and calcium (Ca2+) by microbial-induced calcium precipitation (MICP) after adding fulvic acid (FA). After the addition of 20 mg/L FA, the removal efficiencies of nitrate (NO3--N), Cu2+, CBZ, and Ca2+ reached 100.0 %, 98.7 %, 96.6 %, and 73.6 %, correspondingly and there was no accumulation of nitrite (NO2--N). FA stimulated the growth of strain W12, improved electron transfer activity, and facilitated the conversion of gaseous nitrogen. The research revealed that FA might enhance microbial activity and result in a more dense and porous structure of the biological precipitate. Cu2+ and CBZ were removed by co-precipitation and adsorption. As the initial report of FA promoting MICP to remove complex pollutants, this paper offers a theoretical foundation for NO3--N, Cu2+, CBZ, and Ca2+ remediation in micropolluted water.

A Self-Propelled Linear Piezoelectric Micro-Actuator Inspired by the Movement Patterns of Aquatic Beetles.

The locomotion mechanisms and structural characteristics of insects in nature offer new perspectives and solutions for designing miniature actuators. Inspired by the underwater movement of aquatic beetles, this paper presents a bidirectional self-propelled linear piezoelectric micro-actuator (SLPMA), whose maximum size in three dimensions is currently recognized as the smallest known of the self-propelled piezoelectric linear micro-actuators. Through the superposition of two bending vibration modes, the proposed actuator generates an elliptical motion trajectory at its driving feet. The size was determined as 15 mm × 12.8 mm × 5 mm after finite element analysis (FEA) through modal and transient simulations. A mathematical model was established to analyze and validate the feasibility of the proposed design. Finally, a prototype was fabricated, and an experimental platform was constructed to test the driving characteristics of the SLPMA. The experimental results showed that the maximum no-load velocity and maximum carrying load of the prototype in the forward motion were 17.3 mm/s and 14.8 mN, respectively, while those in the backward motion were 20.5 mm/s and 15.9 mN, respectively.

Robust tetra-armed poly (ethylene glycol)-based hydrogel as tissue bioadhesive for the efficient repair of meniscus tears.

Repair and preservation of the injured meniscus has become paramount in clinical practice. However, the complexities of various clinic stitching techniques for meniscus repair pose challenges for grassroots doctors. Hence, there is a compelling interest in innovative therapeutic strategies such as bioadhesives. An ideal bioadhesive must cure quickly in aqueous and blood environments, bind strongly, endure arthroscopic washing pressures, and degrade appropriately for tissue regeneration. Here, we present a tetra-poly (ethylene glycol) (PEG)-based hydrogel bioadhesive, boasting high biocompatibility, ultrafast gelation, facile injectable operation, and favorable mechanical strength. In view of the synergistic effects of chemical anchor and physical chain entanglement to tightly bind the meniscus, a seamless interface was formed between the surrounding meniscal tissues and hydrogels, enabling the longitudinal tear of the meniscus fused in situ to withstand large tensile force with the adhesive strength of 541.5 ± 31.4 kPa and arthroscopic washout resistance of 29.4 kPa. Superior to existing commercial adhesives, ours allows sutureless application and arthroscopic assistance, without requiring specialized clinical skills. This research is expected to significantly impact our understanding of meniscal healing and ultimately promote a simpler process for achieving functional and structural recovery in torn menisci.

Which technique provides more benefits in return to sports and clinical outcomes after anterior cruciate ligament reconstruction: Double-bundle or single-bundle? A randomized controlled study.

BACKGROUND: The achievement of an optimal return to sport (RTS) has remained a key goal after sports-related injuries, with the ongoing debate on the effectiveness of different surgical approaches for anterior cruciate ligament (ACL) rupture. This study aims to assess clinical outcomes and RTS across various surgical methods, such as anatomical single-bundle reconstruction (ASBR), central-axial single-bundle reconstruction (CASBR), and double-bundle reconstruction (DBR). METHODS: A randomized clinical trial was conducted, comprising 191 patients who underwent ACL rupture. These patients were divided into three groups based on the ACL reconstruction techniques they received (ASBR, CASBR, DBR). Over the 2-year follow-up period, the study assessed RTS through four single-hop tests, isokinetic extension tests, and limb asymmetry indices. Postoperative graft status was determined using the signal-to-noise quotient (SNQ), while knee function was evaluated using the International Knee Documentation Committee 2000 (IKDC-2000) score, Lysholm score, Tegner score, and degree of knee laxity. A binary logistic regression model was developed to forecast the factors influencing ideal RTS. RESULTS: DBR (67.63%) and CASBR (58.00%) exhibited higher RTS passing rates compared to ASBR (30.39%; χ2 = 19.57, P <0.05). Quadriceps strength symmetry in the lower limbs was identified as the key determinant of RTS (χ2 = 17.08, P <0.05). The RTS rate was influenced by SNQs of the graft's tibial site (odds ratio: 0.544) and quadriceps strength of the reconstructed knee joint at 60°/s (odds ratio: 6.346). Notably, the DBR group showed enhanced knee stability, evidenced by superior results in the Lachman test (χ2 = 13.49, P <0.01), objective IKDC-2000 (χ2 = 27.02, P = 0.002), and anterior instability test (χ2 = 9.46, P <0.01). Furthermore, DBR demonstrated superior clinical outcomes based on the Lysholm score (DBR: 89.57 ± 7.72, CASBR: 83.00 ± 12.71, ASBR: 83.21 ± 11.95; F = 10.452, P <0.01) and IKDC-2000 score (DBR: 90.95 ± 7.00, CASBR: 84.64 ± 12.68, ASBR: 83.63 ± 11.41; F = 11.78, P <0.01). CONCLUSION: For patients with ACL rupture, more ideal RTS rate and clinical outcomes were shown in the DBR group than in the ASBR and CASBR groups. Autograft status and quadriceps strength are postively related to RTS. TRIAL REGISTRATION: ClinicalTrials.gov (NCT05400460).

Corneal nerve changes by anti-glaucoma medications examined by in vivo confocal microscopy.

AIM: To evaluate the effects of antiglaucoma eye drops on corneal nerves by in vivo confocal microscopy (IVCM). METHODS: This study comprised 79 patients diagnosed with glaucoma and 16 healthy control individuals. Among the glaucoma patients, 54 were treated with medication, while 25 remained untreated. Central corneal images were evaluated by IVCM, and then ACCMetrics was used to calculate the following parameters: corneal nerve fiber density (CNFD), branch density (CNBD), fiber length (CNFL), total branch density (CTBD), fiber area (CNFA), fiber width (CNFW), and fractal dimension (CNFrD). The correlation between IVCM parameters and drugs was evaluated using non-parametric measurements of Spearman's rank correlation coefficient. RESULTS: The CNFD was reduced in glaucoma groups compared to healthy subjects (P<0.01). Patients using anti-glaucoma medications exhibited poorer confocal parameters compared to untreated patients. As the number of medications and usage count increased, CNFD, CNBD, CNFL, CTBD, CNFA, and CNFrD experienced a decline, while CNFW increased (all P<0.01). For the brinzolamide-therapy group, there was a significant decrease in CNFD and CNFL compared to the other monotherapy groups (P<0.001). In the absence of medication, CNFD in males was lower than that in females (P<0.05). Among patients under medication therapy, CNFD remained consistent between males and females. CONCLUSION: Antiglaucoma eye drops affect the microstructure of corneal nerves. IVCM and ACCMetrics are useful tools that could be used to evaluate the corneal nerve changes.

Exploratory research on the multi-life stages mesh-type model of Caenorhabditis elegans in radiation ecology.

To address the lack of effective dose quantification methods for the model organism Caenorhabditis elegans (C. elegans) in radiation ecology research, this study employs remeshing techniques to develop a comprehensive mesh-type model covering multi-life stages, from embryonic to larval (L1, L2, L3, L4) and adulthood. Using these models, Dose Coefficients (DC) for C. elegans in a soil environment under different exposure conditions (external and internal), material settings, and radioactive nuclides (³H, 6°Co, 9°Sr, 129I, 1³1I, 1³4Cs, 1³7Cs) were calculated with the Monte Carlo toolkit Geant4. The results show that the difference in DC, when C. elegans material is set as either biological material or water, is within 5%. Under external exposure conditions, the impact of life stages on the population's average DC is minimal (with a maximum deviation not exceeding 10%). However, the distribution within the population varied significantly across life stages (under external exposure to 137Cs, the dispersion was 12.02% for adults and a considerably higher 60.30% for larvae). The earlier the life stage, the greater the variability in DC distribution within the C. elegans population. Furthermore, correlation analysis indicates a strong relationship between DC and life stages under internal exposure scenarios. The mesh-type model of C. elegans established in this study provides a valuable tool for radiation ecology research and has potential applications in broader research fields.

A sensitive one-pot ROA assay for rapid miRNA detection.

MicroRNAs (miRNAs) and short RNA fragments (18-25 nt) are crucial biomarkers in biological research and disease diagnostics. However, their accurate and rapid detection remains a challenge, largely due to their low abundance, short length, and sequence similarities. In this study, we report on a highly sensitive, one-step RNA O-circle amplification (ROA) assay for rapid and accurate miRNA detection. The ROA assay commences with the hybridization of a circular probe with the test RNA, followed by a linear rolling circle amplification (RCA) using dUTP. This amplification process is facilitated by U-nick reactions, which lead to an exponential amplification for readout. Under optimized conditions, assays can be completed within an hour, producing an amplification yield up to the microgram level, with a detection limit as low as 0.15 fmol (6 pM). Notably, the ROA assay requires only one step, and the results can be easily read visually, making it user-friendly. This ROA assay has proven effective in detecting various miRNAs and phage ssRNA. Overall, the ROA assay offers a user-friendly, rapid, and accurate solution for miRNA detection. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-024-00140-0.

Multiple effects of microbially induced calcium precipitation on bacteria under different molar volumes of organic pollutants.

Microbially induced calcium precipitation (MICP) has been extensively discussed as a water treatment method. However, the impact of MICP on the selective adsorption of different organic contaminants in industrial wastewater and the metabolism and growth of bacteria has not been elucidated in detail. In this study, by comparing the differences in the metabolism and removal of bacteria by phenol, bisphenol A (BPA), and tetracycline (TC), it was found that bioprecipitates had significant differences in the adsorption capacity of organic pollutants with different molar volumes. Concurrently, bacteria produced more extracellular polymeric substances (EPS) under the influence of organic pollutants, and the self-protection mechanism of bacteria would reduce the amount of gaseous nitrogen. However, the points on the surface of EPS promoted the process of MICP, and MICP encapsulated bacteria to form precipitates to regulate bacteria in water and further improve the removal of carbon and nitrogen in water through biomineralization. This experiment provides new insights into the selective adsorption of bioprecipitates and its multiple effects on bacteria.

Deep learning enhancing guide RNA design for CRISPR/Cas12a-based diagnostics.

Rapid and accurate diagnostic tests are fundamental for improving patient outcomes and combating infectious diseases. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas12a-based detection system has emerged as a promising solution for on-site nucleic acid testing. Nonetheless, the effective design of CRISPR RNA (crRNA) for Cas12a-based detection remains challenging and time-consuming. In this study, we propose an enhanced crRNA design system with deep learning for Cas12a-mediated diagnostics, referred to as EasyDesign. This system employs an optimized convolutional neural network (CNN) prediction model, trained on a comprehensive data set comprising 11,496 experimentally validated Cas12a-based detection cases, encompassing a wide spectrum of prevalent pathogens, achieving Spearman's ρ = 0.812. We further assessed the model performance in crRNA design for four pathogens not included in the training data: Monkeypox Virus, Enterovirus 71, Coxsackievirus A16, and Listeria monocytogenes. The results demonstrated superior prediction performance compared to the traditional experiment screening. Furthermore, we have developed an interactive web server (https://crispr.zhejianglab.com/) that integrates EasyDesign with recombinase polymerase amplification (RPA) primer design, enhancing user accessibility. Through this web-based platform, we successfully designed optimal Cas12a crRNAs for six human papillomavirus (HPV) subtypes. Remarkably, all the top five predicted crRNAs for each HPV subtype exhibited robust fluorescent signals in CRISPR assays, thereby suggesting that the platform could effectively facilitate clinical sample testing. In conclusion, EasyDesign offers a rapid and reliable solution for crRNA design in Cas12a-based detection, which could serve as a valuable tool for clinical diagnostics and research applications.

The meniscotibial ligament does exist: An anatomic and histological description.

Purpose: To describe the anatomical and histological characteristics of the human MTL (meniscotibial ligament) that keeps the meniscus stable and are rarely discussed. Study design: Descriptive laboratory study. Methods: In total, six fresh-frozen adult cadaver knees were dissected, and the dissection protocol were designed by two experienced anatomy professors. The anatomical morphology of MTL was observed. The main anatomical specimens included meniscus, tibial plateau, MTL. The osteotome was used to excise the portion of the tibial plateau, which could obtain the complex including partial meniscus, MTL, and a tibial fragment. A histopathologic study was performed by two experienced pathologists. Results: Macroscopically, the MTL could be divided into two parts: medial meniscotibial ligament (MMTL)and lateral meniscotibial ligament (LMTL). The MMTL is distributed continuously, whereas the LMTL is discontinuous on the tibial plateau. The average length from the tibial attachment of the LMTL to the articular surface was 19 ± 1.0mm (mean ± SD). The average length from the tibial attachment of the MMTL to the articular surface was 10 ± 1.2 mm (mean ± SD). Microscopy of the MTL showed that the MTL is a ligamentous tissue, composed of a network of oriented collagenous fibers. Conclusions: In all knees, the MTL was inserted on the outer edge of the meniscus, attaching to the tibia below the level of articular cartilage, which was key to maintaining the rotational stability of knee and the meniscus in the physiological position on the tibial plateau. Histological analysis of this ligament demonstrated that the MTL is a veritable ligamentous structure, which is made up of collagen type I-expressing fibroblasts. Clinical relevance: This article contributes to the understanding of the anatomical and histological characteristics of the MTL. It is beneficial to promote the development of relevant surgical techniques for the MTL lesion.