Phase-locked µPIV analysis of flow dynamics in a simulated root canal with different laser-activated irrigations.

PURPOSE: To measure the dynamic characteristics of the flow field in a complex root canal model activated by two laser-activated irrigation (LAI) modalities at different activation energy outputs: photon-induced photoacoustic streaming (PIPS) and microshort pulse (MSP). METHODS: A phase-locked micro-scale Particle Image Velocimetry (µPIV) system was employed to characterise the temporal variations of LAI-induced velocity fields in the root canal following a single laser pulse. The wall shear stress (WSS) in the lateral root canal was subsequently estimated from the phase-averaged velocity fields. RESULTS: Both PIPS and MSP were able to generate the 'breath mode' of the irrigant current under all tested conditions. The transient irrigation flush in the root canal peaked at speeds close to 6 m/s. However, this intense flushing effect persisted for only about 2000 µs (or 3% of a single laser-pulse activation cycle). For MSP, the maximum WSS magnitude was approximately 3.08 Pa at an activation energy of E = 20 mJ/pulse, rising to 9.01 Pa at E = 50 mJ/pulse. In comparison, PIPS elevated the WSS to 10.63 Pa at E = 20 mJ/pulse. CONCLUSION: Elevating the activation energy can boost the peak flushing velocity and the maximum WSS, thereby enhancing irrigation efficiency. Given the same activation energy, PIPS outperforms MSP. Additionally, increasing the activation frequency may be an effective strategy to improve irrigation performance further.

Patient-derived Immunocompetent Tumor Organoids: A Platform for Chemotherapy Evaluation in the Context of T-cell Recognition.

Most of the anticancer compounds synthesized by chemists are primarily evaluated for their direct cytotoxic effects at the cellular level, often overlooking the critical role of the immune system. In this study, we developed a patient-derived, T-cell-retaining tumor organoid model that allows us to evaluate the anticancer efficacy of chemical drugs under the synergistic paradigm of antigen-specific T-cell-dependent killing, which may reveal the missed drug hits in the simple cytotoxic assay. We evaluated clinically approved platinum (Pt) drugs and a custom library of twenty-eight PtIV compounds. We observed low direct cytotoxicity of Pt drugs, but variable synergistic effects in combination with immune checkpoint inhibitors (ICIs). In contrast, the majority of PtIV compounds exhibited potent tumor-killing capabilities. Interestingly, several PtIV compounds went beyond direct tumor killing and showed significant immunosynergistic effects with ICIs, outstanding at sub-micromolar concentrations. Among these, Pt-19, PtIV compounds with cinnamate axial ligands, emerged as the most therapeutically potent, demonstrating pronounced immunosynergistic effects by promoting the release of cytotoxic cytokines, activating immune-related pathways and enhancing T cell receptor (TCR) clonal expansion. Overall, this initiative marks the first use of patient-derived immunocompetent tumor organoids to explore and study chemotherapy, advancing their path toward more effective small molecule drug discovery.

Computer-Aided Drug Design Boosts RAS Inhibitor Discovery.

The Rat Sarcoma (RAS) family (NRAS, HRAS, and KRAS) is endowed with GTPase activity to regulate various signaling pathways in ubiquitous animal cells. As proto-oncogenes, RAS mutations can maintain activation, leading to the growth and proliferation of abnormal cells and the development of a variety of human cancers. For the fight against tumors, the discovery of RAS-targeted drugs is of high significance. On the one hand, the structural properties of the RAS protein make it difficult to find inhibitors specifically targeted to it. On the other hand, targeting other molecules in the RAS signaling pathway often leads to severe tissue toxicities due to the lack of disease specificity. However, computer-aided drug design (CADD) can help solve the above problems. As an interdisciplinary approach that combines computational biology with medicinal chemistry, CADD has brought a variety of advances and numerous benefits to drug design, such as the rapid identification of new targets and discovery of new drugs. Based on an overview of RAS features and the history of inhibitor discovery, this review provides insight into the application of mainstream CADD methods to RAS drug design.

Characteristics of the Irrigant Flow in a Simulated Lateral Canal Under Two Typical Laser-Activated Irrigation Regimens.

BACKGROUND AND OBJECTIVES: Photon-induced photoacoustic streaming (PIPS) and shockwave-enhanced emission photoacoustic streaming (SWEEPS) are two promising laser-activated irrigation (LAI) methods for root canal irrigation. Their performance in driving irrigant flush in a complex root canal system will be evaluated by microscale particle image velocimetry (µPIV) measurement and will be compared with that of ultrasonic-activated irrigation (UAI). STUDY DESIGN/MATERIALS AND METHODS: A µPIV system with 7 µm fluorescent tracer particles was adopted to measure two-dimensional (2D) velocity fields around the junction region, with a size of 1.8 mm × 1.5 mm, between one main canal and one lateral canal in an endodontic training block, which was driven by SWEEPS (Er:YAG laser) operating at 15 Hz and 20 mJ. The flow field driven by PIPS (Er:YAG laser) at the same frequency and energy, as well as by UAI (with non-cutting insert) operating at 40% unit power, was also measured for a direct comparison. RESULTS: It was found that both SWEEPS and PIPS can activate a so-called "breath mode" during the irrigation. Namely, the induced irrigant flush presented a back-and-forth oscillation along both the main canal and the lateral one. The maximum flow speed in the lateral canal was observed to be up to 10 m/s in the SWEEPS modality, while reduced to around 7 m/s in the PIPS modality. The penetration length in the lateral canal in both modalities was estimated to be larger than 1 mm. In comparison, the flow field induced by UAI was characterized as irregular vortical structures, the maximum flow speed in the lateral canal was 0.15 m/s and significantly lower than LAI (P < 0.01), and the penetration length was less than 300 µm and lower than LAI (P < 0.05). CONCLUSION: Compared to UAI, PIPS, and SWEEPS are more capable of delivering the irrigant deeper into the lateral canal. Furthermore, the back-and-forth flush in the breath mode is ideal for removing debris during irrigation. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

An Integrated Multi-Sensor Network for Adaptive Grasping of Fragile Fruits: Design and Feasibility Tests.

Secure grasping of fragile fruits and other agricultural products without potential slip and damage is still a challenge due to the size and shape varying, bruise susceptible, as well as hardness changing during fruit and vegetable maturation. In the robotic grasping process, the mechanical damage mainly depends upon the aggressiveness of the gripper and the sensitivity of the product to the damage. In this study, a flexible gripper integrated with multi-sensor network is designed and tested. The network proposed includes three kinds of sensors that enable the gripper to grasp various products with the sense of touch and visual perception. Particular attention has been attached to the sensors applied between the fingers, and this makes sensing and grasping capabilities improved. To create an accurate grasping system, a grasping algorithm and the force control model are proposed for any bending state based on Cosserat theory. The boundary detection is included in the grasping algorithm, detecting the shape edge by some certain point calculation. The created grasping system guarantees mechanical compliance by evaluating and adjusting the finger status including force, angle, and direction. Multi-group tests have been done on grasping several objects of different sizes and materials in daily life. The relationship between force, bending, and surface material is also analyzed and compared under different conditions. The numerical comparisons related to the measurement error are analyzed based on their standard deviations. Experimental results indicate that this flexible manipulator with proposed system and strategy has better grasping ability for fragile fruits with its good flexibility and dexterity.

Facile Preparation of CO2 -Responsive Polymer Nano-Objects via Aqueous Photoinitiated Polymerization-Induced Self-Assembly (Photo-PISA).

Carbon dioxide (CO2 )-responsive polymer nano-objects are prepared by photoinitiated reversible addition-fragmentation chain transfer dispersion polymerization of 2-hydroxypropyl methacrylate and 2-(dimethylamino)ethyl methacrylate (DMAEMA) in water at room temperature using a poly(poly(ethylene glycol) methyl ether methacrylate) macromolecular chain transfer agent. Kinetic studies confirm that full monomer conversions are achieved in all cases within 10 min of visible-light irradiation (405 nm, 0.5 mW cm-2 ). The effect of DMAEMA on the polymerization is studied in detail, and pure higher order morphologies (worms and vesicles) are prepared by this particular formulation. Finally, CO2 -responsive property of the obtained vesicles is investigated by dynamic light scattering, visual appearance, and transmission electron microscope.

Low-Temperature Synthesis of Thermoresponsive Diblock Copolymer Nano-Objects via Aqueous Photoinitiated Polymerization-Induced Self-Assembly (Photo-PISA) using Thermoresponsive Macro-RAFT Agents.

Photoinitiated reversible addition-fragmentation chain transfer (RAFT) dispersion polymerization of 2-hydroxypropyl methacrylate is conducted in water at low temperature using thermoresponsive copolymers of 2-(2-methoxyethoxy) ethyl methacrylate and oligo(ethylene glycol) methacrylate (Mn = 475 g mol(-1) ) as the macro-RAFT agent. Kinetic studies confirm that quantitative monomer conversion is achieved within 15 min of visible-light irradiation (405 nm, 0.5 mW cm(-2) ), and good control is maintained during the polymerization. The polymerization can be temporally controlled by a simple "ON/OFF" switch of the light source. Finally, thermoresponsive diblock copolymer nano-objects with a diverse set of complex morphologies (spheres, worms, and vesicles) are prepared using this particular formulation.

Prediction of immune infiltration and prognosis for patients with urothelial bladder cancer based on the DNA damage repair-related genes signature.

Objectives: To analyze the correlations between the expression and effect of DNA damage repair genes and the immune status and clinical outcomes of urothelial bladder cancer (BLCA) patients. In addition, we evaluate the efficacy and value of utilizing the DNA damage repair genes signature as a prognosis model for BLCA. Methods: Two subtype groups (C1 and C2) were produced based on the varied expression of DNA damage repair genes. Significantly differentiated genes and predicted enriched gene pathways were obtained between the two subtypes. Seven key genes were obtained from the DNA damage repair-related genes and a 7-gene signature prognosis model was established based on the key genes. The efficacy and accuracy of this model in prognosis prediction was evaluated and verified in two independent databases. Also, the difference in biological functions, drug sensitivity, immune infiltration and affinity between the high-risk group and low-risk group was analyzed. Results: The DNA damage repair gene signature could significantly differentiate the BLCA into two molecular subgroups with varied genetic expression and enriched gene pathways. Seven key genes were screened out from the 232 candidate genes for prognosis prediction and a 7-gene signature prognosis model was established based on them. Two independent patient cohorts (TCGA cohort and GEO cohort) were utilized to validate the efficacy of the prognosis model, which demonstrated an effective capability to differentiate and predict the overall survival of BLCA patients. Also, the high-risk group and low-risk group derived from the 7-gene model exhibited significantly differences in drug sensitivity, immune infiltration status and biological pathways enrichment. Conclusions: Our established 7-gene signature model based on the DNA damage repair genes could serve as a novel prognosis predictive tool for BLCA. The differentiation of BLCA patients based on the 7-gene signature model may be of great value for the appropriate selection of specific chemotherapy agents and immune-checkpoint blockade therapy administration.

ALOX5 deficiency contributes to bladder cancer progression by mediating ferroptosis escape.

Ferroptosis is an iron-dependent form of regulated cell death driven by the lethal lipid peroxides. Previous studies have demonstrated that inducing ferroptosis holds great potential in cancer therapy, especially for patients with traditional therapy failure. However, cancer cells can acquire ferroptosis evasion during progression. To date, the therapeutic potential of inducing ferroptosis in bladder cancer (BCa) remains unclear, and whether a ferroptosis escape mechanism exists in BCa needs further investigation. This study verified that low pathological stage BCa cells were highly sensitive to RSL3-induced ferroptosis, whereas high pathological stage BCa cells exhibited obviously ferroptosis resistance. RNA-seq, RNAi-mediated loss-of-function, and CRISPR/Cas9 experiments demonstrated that ALOX5 deficiency was the crucial factor of BCa resistance to ferroptosis in vitro and in vivo. Mechanistically, we found that ALOX5 deficiency was regulated by EGR1 at the transcriptional level. Clinically, ALOX5 expression was decreased in BCa tissues, and its low expression was associated with poor survival. Collectively, this study uncovers a novel mechanism for BCa ferroptosis escape and proposes that ALOX5 may be a valuable therapeutic target and prognostic biomarker in BCa treatment.

Analysis and identification of the necroptosis landscape on therapy and prognosis in bladder cancer.

Bladder cancer (BLCA) is one of the most common malignant tumors of the urinary system, but the current therapeutic strategy based on chemotherapy and immune checkpoint inhibitor (ICI) therapy cannot meet the treatment needs, mainly owing to the endogenous or acquired apoptotic resistance of cancer cells. Targeting necroptosis provides a novel strategy for chemotherapy and targeted drugs and improves the efficacy of ICIs because of strong immunogenicity of necroptosis. Therefore, we systemically analyzed the necroptosis landscape on therapy and prognosis in BLCA. We first divided BLCA patients from The Cancer Genome Atlas (TCGA) database into two necroptosis-related clusters (C1 and C2). Necroptosis C2 showed a significantly better prognosis than C1, and the differential genes of C2 and C1 were mainly related to the immune response according to GO and KEGG analyses. Next, we constructed a novel necroptosis-related gene (NRG) signature consisting of SIRT6, FASN, GNLY, FNDC4, SRC, ANXA1, AIM2, and IKBKB to predict the survival of TCGA-BLCA cohort, and the accuracy of the NRG score was also verified by external datasets. In addition, a nomogram combining NRG score and several clinicopathological features was established to more accurately and conveniently predict the BLCA patient's survival. We also found that the NRG score was significantly related to the infiltration levels of CD8 T cells, NK cells, and iDC cells, the gene expression of CTLA4, PD-1, TIGIT, and LAG3 of TME, and the sensitivity to chemotherapy and targeted agents in BLCA patients. In conclusion, the NRG score has an excellent performance in evaluating the prognosis, clinicopathologic features, tumor microenvironment (TME), and therapeutic sensitivity of BLCA patients, which could be utilized as a guide for chemotherapy, ICI therapy, and combination therapy.