Dual-Stimuli Regulation of DNAzyme Cleavage Reaction by Coordination-Driven Nanoprobes for Cancer Cell Imaging.

Endowing current artificial chemical reactions (ACRs) with high specificity and intricate activation capabilities is crucial for expanding their applications in accurate bioimaging within living cells. However, most of the reported ACR-based evaluations relied on either single biomarker stimuli or dual activators without obvious biological relevance, still limiting their accuracy and fidelity. Herein, taking the metal-ion-dependent DNAzyme cleavage reaction as a model ACR, two regulators, glutathione (GSH) and telomerase (TE) activated DNAzyme cleavage reactions, were exploited for precise discrimination of cancerous cells from normal cells. DNA probe was self-assembled into the ZIF-90 nanoparticle framework to construct coordination-driven nanoprobes. This approach enhances the stability and specificity of tumor imaging by utilizing biomarkers associated with rapid tumor proliferation and those commonly overexpressed in tumors. In conclusion, the research not only paves the way for new perspectives in cell biology and pathology studies but also lays a solid foundation for the advancement of biomedical imaging and disease diagnostic technologies.

Understanding the Role of Biofilms in Acute Recurrent Tonsillitis through 3D Bioprinting of a Novel Gelatin-PEGDA Hydrogel.

Acute recurrent tonsillitis is a chronic, biofilm-related infection that is a significant burden to patients and healthcare systems. It is often treated with repeated courses of antibiotics, which contributes to antimicrobial resistance. Studying biofilms is key to understanding this disease. In vitro modelling using 3D bioprinted hydrogels is a promising approach to achieve this. A novel gelatin-PEGDA pseudomonas fluorescens-laden bioink was developed and bioprinted in a 3D hydrogel construct fabricated using computer-aided design to mimic the tonsillar biofilm environment. The bioprinted constructs were cultured at 37 °C in lysogeny broth for 12 days. Bacterial growth was assessed by spectrophotometry. Cellular viability analysis was conducted using optical fluorescence microscopy (FDA/PI staining). A biocompatible 3D-printed bacteria-laden hydrogel construct was successfully fabricated. Bacterial growth was observed using optical fluorescence microscopy. A live/dead cellular-staining protocol demonstrated bacterial viability. Results obtained after the 12-day culture period showed higher bacterial growth in the 1% gelatin concentration construct compared to the 0% control. This study demonstrates the first use of a bacteria-laden gelatin-PEGDA hydrogel for biofabrication of a 3D-printed construct designed to model acute recurrent tonsillitis. Initiating a study with clinically relevant ex vivo tonsil bacteria will be an important next step in improving treatment of this impactful but understudied disease.

Orthogonally Sequential Activation of Self-Powered DNAzymes Cascade for Reliable Monitoring of mRNA in Living Cells.

Ratiometric imaging of tumor-related mRNA is significant, yet spatiotemporally resolved regulation on the ratiometric signals to avoid non-specific activation in the living cells remains challenging. Herein, orthogonally sequential activation of concatenated DNAzyme circuits is, first, developed for Spatio Temporally regulated Amplified and Ratiometric (STAR) imaging of TK1 mRNA inside living cells with enhanced reliability and accuracy. By virtue of the synthesized CuO/MnO2 nanosheets, orthogonally regulated self-powered DNAzyme circuits are operated precisely in living cells, sequentially activating two-layered DNAzyme cleavage reactions to achieve the two ratiometric signal readouts successively for reliable monitoring of low-abundance mRNA in living cells. It is found that the ratiometric signals can only be derived from mRNA over-expressed tumor cells, also irrespective of probes' delivery concentration. The presented approach could provide new insight into orthogonally regulated ratiometric systems for reliable imaging of specific biomarkers in living cells, benefiting disease precision diagnostics.

Autonomous operation of 3D DNA walkers in living cells for microRNA imaging.

Three dimensional (3D) DNA walkers hold great potential in serving as an ideal candidate for signal transduction and amplification in bio-assays. However, the autonomous operation of 3D DNA walkers inside living cells is still few and far between, which could be attributed to the lack of suitable driving forces and moderate efficiency in terms of the cellular uptake of such complex 3D DNA components. Herein, a newly updated autonomously operated and highly integrated 3D DNA walker on Au nanoparticles (Au NPs)/zeolitic imidazolate framework-8 (ZIF-8) was activated in a tumor microenviroment and its signal amplified assay capability in living cells was demonstrated using miRNA as a sensing model biomolecule. Specifically, we assembled a 3D DNA motor, including Zn2+-dependent DNAzyme and substrates on the AuNPs grafted on ZIF-8. After being delivered into a living cell, ZIF-8 was efficiently degraded in the tumor microenvironment (low pH value), locally releasing the Zn2+ and DNA motor. Then, a self-sufficient DNA motor autonomously performed the bio-analytical task of imaging miRNA-10b, with a low detection limit of 34 pM. Also, such self-sufficient 3D walkers allowed real-time imaging of MDA-MB-231 cells by intracellular operation. This method demonstrates the self-sufficient 3D DNA motor's bioanalytical application in living cells which may inspire various other biological applications including gene delivery, therapy, etc.

Femoral Stress Changes after Total Hip Arthroplasty with the Ribbed Prosthesis: A Finite Element Analysis.

BACKGROUND: A total hip reconstruction is related to the stress distribution throughout the prosthesis, cement, and femur. Researches on reducing the stress in all components to minimize the risk of failure are of great significance. The objective of our study was to determine the biomechanical variation in overall femoral stress and periprosthetic femoral stress distribution after implantation with the Ribbed anatomic prosthesis. METHODS: Three-dimensional finite element models of intact femur and Ribbed prosthesis were developed according to the morphology, while the hip joint loading and the strength of related muscles were applied in the models. The overall stress changes of the intact femur before and after the implantation were analyzed, and the periprosthetic stress distribution especially in the proximal region of the femur was quantified. RESULTS: As a result, the overall stress pattern of the femur did not change after the implantation compared with the intact femur. The region of peak stress value was located in the middle and lower segments of the full length femur, but the stress value level decreased. The final prosthesis resulted in a significant decrease in the equivalent stress level of the periprosthetic bone tissue, and the most severe area appeared at the endmost posterior quadrant. The stress shielding ratio of the Ribbed prosthesis was 71.6%. The stress value level gradually increased towards the distal part of the prosthesis and recovered to physiological level at the end of the prosthesis. CONCLUSIONS: The Ribbed prosthesis can cause significant stress shielding effect in the proximal femur. These results may help optimize prosthetic design to reduce stress shielding effect and improve clinical outcomes.

In Situ Cation Exchange Generated ZnS-Ag2S Nanoparticles for Photothermal Detection of Transcription Factor.

The photothermal reagent (PTA)-mediated point-of-care detection of disease biomarkers using thermometers as readout has attracted increasing attention, but complex modification or generation process of PTAs still limited their further applications. Herein, we report a photothermal detection platform in which the target recognition triggered the in situ generation of the PTA and ZnS-Ag2S nanoparticles (NPs) by one-step autonomous cation exchange reaction (ACER). As a proof of concept, NF-kB1, a kind of disease biomarker, was used to demonstrate the photothermal detection platform. First, a triplex-like DNA structure containing the recognition part of NF-kB1 and cytosine-Ag+-cytosine (C-Ag+-C) unit was designed. With the recognition of NF-kB1, abundant C-Ag+-C units were released to take ACER with the prepared ZnS NPs. In addition, because of the intrinsic detecting limitation of the thermometer, hybridization chain reaction (HCR) was introduced to load more Ag+ for the enhancement of the photothermal signal. After the irradiation of 808 nm laser toward the generated ZnS-Ag2S NPs dispersions, its temperature increased with the increased concentration of NF-kB1 in the range from 10 to 1000 nM with a detection limit of 2.31 nM. The signal-amplifying role of HCR was confirmed by the control experiment. This platform of in situ generation of PTA not only integrated the recognition process with detection but also avoided the complex process of modification or generation, which has remarkable potential to be extended for the detections of other different disease biomarkers.

Urinary Metabolites Associated with Blood Pressure on a Low- or High-Sodium Diet.

Dietary salt intake has significant effects on arterial blood pressure and the development of hypertension. Mechanisms underlying salt-dependent changes in blood pressure remain poorly understood, and it is difficult to assess blood pressure salt-sensitivity clinically. Methods: We examined urinary levels of metabolites in 103 participants of the Dietary Approaches to Stop Hypertension (DASH)-Sodium trial after nearly 30 days on a defined diet containing high sodium (targeting 150 mmol sodium intake per day) or low sodium (50 mmol per day). Targeted chromatography/mass spectrometry analysis was performed in 24 h urine samples for 47 amino metabolites and 10 metabolites related to the tricarboxylic acid cycle. The effect of an identified metabolite on blood pressure was examined in Dahl salt-sensitive rats. Results: Urinary metabolite levels improved the prediction of classification of blood pressure salt-sensitivity based on race, age and sex. Random forest and generalized linear mixed model analyses identified significant (false discovery rate <0.05) associations of 24 h excretions of ß-aminoisobutyric acid, cystine, citrulline, homocysteine and lysine with systolic blood pressure and cystine with diastolic blood pressure. The differences in homocysteine levels between low- and high-sodium intakes were significantly associated with the differences in diastolic blood pressure. These associations were significant with or without considering demographic factors. Treatment with ß-aminoisobutyric acid significantly attenuated high-salt-induced hypertension in Dahl salt-sensitive rats. Conclusion: These findings support the presence of new mechanisms of blood pressure regulation involving metabolic intermediaries, which could be developed as markers or therapeutic targets for salt-sensitive hypertension.