A High-Temperature Superconducting Bandpass Dual-Mode Filter with Tunable Relative Bandwidth and Center Frequency.

This study proposes a high-temperature superconducting (HTS) bandpass filter with a continuously tunable bandwidth and center frequency. The proposed filter combines several gallium arsenide varactors and a dual-mode resonator (DMR). The even and odd modes of the DMR can be tuned simultaneously using a single bias voltage. The capacitive value of varactors in the circuit is tuned continuously under continuous voltage and frequency tunability. External couplings and the interstage can be realized using an interdigital coupling structure; a fixed capacitor is added to the feeder to improve its coupling strength. A low-insertion loss within the band is obtained using HTS technology. Additionally, the proposed filter is etched on a 0.5 mm-thick MgO substrate and combined with YBCO thin films for demonstration. For the as-fabricated device, the tuning frequency range of 1.22~1.34 GHz was 9.4%; the 3-dB fractional bandwidth was 12.95~17.39%, and the insertion loss was 2.28~3.59 dB. The simulation and experimental measurement results were highly consistent.

Deep-learning-based 3D cellular force reconstruction directly from volumetric images.

The forces exerted by single cells in the three-dimensional (3D) environments play a crucial role in modulating cellular functions and behaviors closely related to physiological and pathological processes. Cellular force microscopy (CFM) provides a feasible solution for quantifying mechanical interactions, which usually regains cellular forces from deformation information of extracellular matrices embedded with fluorescent beads. Owing to computational complexity, traditional 3D-CFM is usually extremely time consuming, which makes it challenging for efficient force recovery and large-scale sample analysis. With the aid of deep neural networks, this study puts forward a novel, data-driven 3D-CFM to reconstruct 3D cellular force fields directly from volumetric images with random fluorescence patterns. The deep-learning-based network is established through stacking deep convolutional neural networks (DCNN) and specific function layers. Some necessary physical information associated with constitutive relation of extracellular matrix material is coupled to the data-driven network. The mini-batch stochastic-gradient-descent and back-propagation algorithms are introduced to ensure its convergence and training efficiency. The networks not only have good generalization ability and robustness but also can recover 3D cellular forces directly from the input fluorescence image pairs. Particularly, the computational efficiency of the deep-learning-based network is at least one to two orders of magnitude higher than that of traditional 3D-CFM. This study provides a novel scheme for developing high-performance 3D-CFM to quantitatively characterize mechanical interactions between single cells and surrounding extracellular matrices, which is of vital importance for quantitative investigations in biomechanics and mechanobiology.

Injectable GelMA Cryogel Microspheres for Modularized Cell Delivery and Potential Vascularized Bone Regeneration.

Cell therapeutics hold tremendous regenerative potential and the therapeutic effect depends on the effective delivery of cells. However, current cell delivery carriers with unsuitable cytocompatibility and topological structure demonstrate poor cell viability during injection. Therefore, porous shape-memory cryogel microspheres (CMS) are prepared from methacrylated gelatin (GelMA) by combining an emulsion technique with gradient-cooling cryogelation. Pore sizes of the CMS are adjusted via the gradient-cooling procedure, with the optimized pore size (15.5 ± 6.0 µm) being achieved on the 30-min gradient-cooled variant (CMS-30). Unlike hydrogel microspheres (HMS), CMS promotes human bone marrow stromal cell (hBMSC) and human umbilical vein endothelial cell (HUVEC) adhesion, proliferated with high levels of stemness for 7 d, and protects cells during the injection process using a 26G syringe needle. Moreover, CMS-30 enhances the osteogenic differentiation of hBMSCs in osteoinductive media. CMS can serve as building blocks for delivering multiple cell types. Here, hBMSC-loaded and HUVEC-loaded CMS-30, mixed at a 1:1 ratio, are injected subcutaneously into nude mice for 2 months. Results show the development of vascularized bone-like tissue with high levels of OCN and CD31. These findings indicate that GelMA CMS of a certain pore size can effectively deliver multiple cells to achieve functional tissue regeneration.

Electrodeformation-Based Biomechanical Chip for Quantifying Global Viscoelasticity of Cancer Cells Regulated by Cell Cycle.

Mechanical phenotypes of cells are found to hold vital clues to reveal cellular functions and behaviors, which not only has great physiological significance but also is crucial for disease diagnosis. To this end, we developed a set of electrodeformation-based biomechanical microchip assays to quantify mechanical phenotypes on the single-cell level. By investigating the spatiotemporal dynamics of cancer cells driven by dielectrophoresis forces, we captured the key global viscoelastic indexes including cellular elasticity, viscosity, and transition time that was defined as the ratio of the transient viscosity and elasticity, simultaneously, and thus explored their intrinsic correlation with cell cycle progression. Our results showed that both global elasticity and viscosity have a significant periodic variation with cell cycle progression, but the transition time remained unchanged in the process, indicating that it might be an intrinsic property of cancer cells that is independent of the cell cycle and the type of cell in the experiments. Further, we investigated the molecular mechanism regulating cellular viscoelastic phenotypes on the biomechanical chips through intracellular cytoskeletal perturbation assays. These findings, together with the electrodeformation-based microchip technique, not only reveal the relation between mechanical phenotypes of cancer cells and cell cycle progression but also provide a platform for implementing multi-index mechanical phenotype assays associated with cancer cell cycles in the clinic.

Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection.

Electrotransfection is a widely used method for delivering genes into cells with electric pulses. Although different hypotheses have been proposed, the mechanism of electrotransfection remains controversial. Previous studies have indicated that uptake and intracellular trafficking of plasmid DNA (pDNA) are mediated by endocytic pathways, but it is still unclear which pathways are directly involved in the delivery. To this end, the present study investigated the dependence of electrotransfection on macropinocytosis. Data from the study demonstrated that electric pulses induced cell membrane ruffling and actin cytoskeleton remodeling. Using fluorescently labeled pDNA and a macropinocytosis marker (i.e., dextran), the study showed that electrotransfected pDNA co-localized with dextran in intracellular vesicles. Furthermore, electrotransfection efficiency could be decreased significantly by reducing temperature or treatment of cells with a pharmacological inhibitor of Rac1 and could be altered by changing Rac1 activity. Taken together, the findings suggested that electrotransfection of pDNA involved Rac1-dependent macropinocytosis.

A signal-on ratiometric fluorometric heparin assay based on the direct interaction between amino-modified carbon dots and DNA.

Amino-modified carbon dots (C-dots) with positively charged surface were prepared. They display strong blue fluorescence and are shown to act as quenchers of the green fluorescence of FAM-labeled ssDNA such as the F-probe used in this work that was immobilized on the C-dots. On the addition of highly negatively charged heparin (Hep), it will interact with the C-dots and displace the F-probe from C-dots. Once the F-probe is displaced by Hep, its green fluorescence is restored. The intrinsic blue fluorescence of the C-dots remains stable after addition of Hep. Thus, a signal-on ratiometric fluorometric assay was developed for the ultra-sensitive detection of Hep. The underlying mechanisms of quenching and recovery are discussed. Under optimized conditions, the recovery of the ratiometric fluorescence of the system composed of C-dots and quenched F-probe is proportional to the Hep concentration in the range of 0.01-2.0 µg·mL-1 (= 0.00125-0.25 U·mL-1). The method was successfully applied to the determination of Hep in spiked serum samples. Graphical abstract Schematic of a signal-on ratiometric fluorometric method for the ultra-sensitive detection of heparin on the basis of the displacement and fluorescence enhancement of adsorbed FAM-labeled ssDNA from amino-modified carbon dots (C-dots) by heparin.

Substrate stiffness promotes latent TGF-ß1 activation in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) was usually coupled with increased stiffness of the extracellular matrix (ECM) and elevated level of transforming growth factor-ß1 (TGF-ß1). However, the mechanism by which substrate rigidity modulated TGF-ß1 signaling transduction remained unknown. This paper investigated the molecular mechanism of how matrix stiffness regulating TGF-ß1 signaling in HCC cells. By means of stiffness tunable collagen I-coated polyacrylamide (PA) gels, we found that the expressions of ß1 integrin, p-FAK Y397 and p-Smad2 upregulated on stiffer gels as well as the content of TGF-ß1 in culture media of HCC cells, which were inhibited by RGD blocking peptides, Y-27632 (ROCK inhibitor) or Blebbistatin (myosin II inhibitor). Cellular traction force was also significantly higher when plated on stiffer substrates but dramatically decreased after treatment with Y-27632 or Blebbistatin. Furthermore, the upregulation of p-Smad2 in the HCC cells on stiffer PA gels induced by exogenetic latent TGF-ß1 was downregulated in the presence of RGD peptides. The nuclear translocation of Smad2 induced by latent TGF-ß1 was inhibited by Y-27632 or Blebbistatin. Our results suggested that the extracellular matrix stiffness regulated latent TGF-ß1 activation by cytoskeletal tension in HCC cells, showing that matrix stiffness was a key regulator involving the TGF-ß1 activity in HCC cells. The current study presented a mechanism of how hepatocirrhosis developed into liver cancer.

Integrated water resources management for emergency situations: A case study of Macau.

Integrated urban water management (IUWM) is a useful tool that can be used to alleviate water resource shortages in developing regions like Macau, where 98% of the raw water comes from mainland China. In Macau, scarce water resources deteriorate rapidly in emergency situations, such as accidental chemical spills upstream of the supply reservoir or salty tides. During these times, only the water from the two freshwater reservoirs in Macau can be used. In this study, we developed urban water management optimization models that integrated the raw water supply from the two reservoirs with various proposed governmental policies (wastewater reuse, rainwater collection, and water saving). We then determined how various water resource strategies would influence the urban water supply in Macau in emergency situations. Our results showed that, without imported raw water, the water supply from only the two Macau reservoirs would last for 7.95days. However, when all the government policies were included in the model, the supply could be extended to 13.79days. Out of the three non-conventional water resources, wastewater reuse is the most beneficial for increasing the Macau water supply, and rainwater collection also has great potential.

Mechanical analysis of rat trabecular meshwork.

Stiffness of trabecular meshwork (TM) may play an important role in regulating outflow resistance in healthy and glaucomatous eyes. However, the current techniques for stiffness measurement can only be applied to TM dissected from human donor or large animal eyes. It is a challenge to measure TM stiffness in mouse/rat eyes because of their smaller sizes and the delicate nature of TM dissection. To this end, a new technique was developed to determine the stiffness of rat TM using atomic force microscopy (AFM). In the study, rat eyes were enucleated immediately after death and perfused with a tracer (Evans blue) for 40 min. Then, the anterior segment was dissected and flat-mounted on a Petri dish with TM facing upwards. An AFM probe with a gold-coated colloid tip was used to sequentially indent the corneal, TM, and uveoscleral tissues. Assuming these tissues to be neo-Hookean materials, the indentation data were analyzed with a newly developed mathematical model to calculate the apparent initial Young's moduli (E0)(app). The geometric mean & SE of (E0)(app) were 162 Pa & 1.2 (n = 13) for TM and 6189 Pa & 1.4 (n = 11) for cornea; and the difference was statistically significant (p < 0.01). The technique established in this study allows the use of rat eye as a potential model for investigation of TM stiffness and its influences on outflow resistance. Future studies may also utilize this technique to evaluate mechanisms of TM stiffness change caused by aging, outflow dysfunction, pathogenesis of glaucoma, and drug treatment.

Water Impalement Resistance and Drag Reduction of the Superhydrophobic Surface with Hydrophilic Strips.

Superhydrophobic surfaces (SHS) offer versatile applications by trapping an air layer within microstructures, while water jet impact can destabilize this air layer and deactivate the functions of the SHS. The current work presents for the first time that introducing parallel hydrophilic strips to SHS (SHS-s) can simultaneously improve both water impalement resistance and drag reduction (DR). Compared with SHS, SHS-s demonstrates a 125% increase in the enduring time against the impact of water jet with velocity of 11.9 m/s and a 97% improvement in DR at a Reynolds number of 1.4 × 104. The key mechanism lies in the enhanced stability of the air layer due to air confinement by the adjacent three-phase contact lines. These lines not only impede air drainage through the surface microstructures during water jet impact, entrapping the air layer to resist water impalement, but also prevent air floating up due to buoyancy in Taylor-Couette flow, ensuring an even spread of the air layer all over the rotor, boosting DR. Moreover, failure modes of SHS under water jet impact are revealed to be related to air layer decay and surface structure destruction. This mass-producible structured surface holds the potential for widespread use in DR for hulls, autonomous underwater vehicles, and submarines.

Corrigendum to "Anti-tumor effect and mechanism of the total biflavonoid extract from S doederleinii on human cervical cancer cells in vitro and in vivo" [Heliyon 10(2) January 2024 e24778].

[This corrects the article DOI: 10.1016/j.heliyon.2024.e24778.].

Anti-tumor effect and mechanism of the total biflavonoid extract from S doederleinii on human cervical cancer cells in vitro and in vivo.

In this study, the therapeutic effect and possible mechanism of the total biflavonoid extract of Selaginella doederleinii Hieron (SDTBE) against cervical cancer were originally investigated in vitro and in vivo. First, the inhibition of SDTBE on proliferation of cervical cancer HeLa cells was evaluated, followed by morphological observation with AO/EB staining, Annexin V/PI assay, and autophagic flux monitoring to evaluate the possible effect of SDTBE on cell apoptosis and autophagy. Cell cycle, as well as mitochondrial membrane potential (ΔÑ°m), was detected with flow cytometry. Further, the apoptosis related protein expression and the autophagy related gene LC3 mRNA transcription level were analyzed by Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR), respectively. Finally, the anti-cervical cancer effect of the SDTBE was also validated in vivo in HeLa cells grafts mice. As results, SDTBE inhibited HeLa cells proliferation with the IC50 values of 49.05 ± 6.76 and 44.14 ± 4.75 µg/mL for 48 and 72 h treatment, respectively. The extract caused mitochondrial ΔÑ° loss, induced cell apoptosis by upregulating Bax, downregulating Bcl-2, activating Caspase-9 and Caspase-3, promoting cell autophagy and blocking the cell cycle in G0/G1 phase. Furthermore, 100, 200, and 300 mg/kg SDTBE suppressed the growth of HeLa cells xenografts in mice with the mean inhibition rates, 25.3 %, 57.5 % and 62.9 %, respectively, and the change of apoptosis related proteins and microvascular density was confirmed in xenografts by immunohistochemistry analysis. The results show that SDTBE possesses anti-cervical cancer effect, and the mechanism involves in activating Caspase-dependent mitochondrial apoptosis pathway.

Extracellular Matrix Rigidities Regulate the Tricarboxylic Acid Cycle and Antibiotic Resistance of Three-Dimensionally Confined Bacterial Microcolonies.

As a major cause of clinical chronic infection, microbial biofilms/microcolonies in host tissues essentially live in 3D-constrained microenvironments, which potentially modulate their spatial self-organization and morphodynamics. However, it still remains unclear whether and how mechanical cues of 3D confined microenvironments, for example, extracellular matrix (ECM) stiffness, exert an impact on antibiotic resistance of bacterial biofilms/microcolonies. With a high-throughput antibiotic sensitivity testing (AST) platform, it is revealed that 3D ECM rigidities greatly modulate their resistance to diverse antibiotics. The microcolonies in 3D ECM with human tissue-specific rigidities varying from 0.5 to 20 kPa show a ≈2-10 000-fold increase in minimum inhibitory concentration, depending on the types of antibiotics. The authors subsequently identified that the increase in 3D ECM rigidities leads to the downregulation of the tricarboxylic acid (TCA) cycle, which is responsible for enhanced antibiotic resistance. Further, it is shown that fumarate, as a potentiator of TCA cycle activity, can alleviate the elevated antibiotic resistance and thus remarkably improve the efficacy of antibiotics against bacterial microcolonies in 3D confined ECM, as confirmed in the chronic infection mice model. These findings suggest fumarate can be employed as an antibiotic adjuvant to effectively treat infections induced by bacterial biofilms/microcolonies in a 3D-confined environment.

Quaternized carbon dots with enhanced antimicrobial ability towards Gram-negative bacteria for the treatment of acute peritonitis caused by E. coli.

Infections caused by Gram-negative bacteria still pose a clinical challenge. Although nanomaterials have been developed for antibacterial treatments, a systematic evaluation of the mechanisms and intervention models of antibacterial materials toward Gram-negative bacteria is still lacking. Herein, antibacterial quaternized carbon dots (QCDs) were synthesized via a one-step melting method using anhydrous citric acid and diallyl dimethyl ammonium chloride (DDA). The QCDs exhibited effective broad-spectrum antibacterial activity and enhanced inhibitory ability towards Gram-negative bacteria. The antibacterial mechanism of the QCDs with respect to Gram-negative bacteria was investigated through the characterization of bacterial morphology changes, the absorption modes of the QCDs on bacteria, and the potential generation of reactive oxygen species by the QCDs. The QCDs showed low toxicity in different cells, and did not cause hemolysis. The QCDs were administered via intraperitoneal injection to treat acute peritonitis in mice infected with E. coli. Routine blood examination, magnetic resonance imaging, and pathological analysis were undertaken and it was found that, similar to the positive control group treated with gentamicin sulfate, the QCDs exhibited a therapeutic effect that eliminated infection and inflammation. This study explores a controllable synthetic strategy for the synthesis of active carbon dots with antibacterial activity, a material that is a promising candidate for new treatments of Gram-negative bacterial infections.

Catch bond-inspired hydrogels with repeatable and loading rate-sensitive specific adhesion.

Biological receptor-ligand adhesion governed by mammalian cells involves a series of mechanochemical processes that can realize reversible, loading rate-dependent specific interfacial bonding, and even exhibit a counterintuitive behavior called catch bonds that tend to have much longer lifetimes when larger pulling forces are applied. Inspired by these catch bonds, we designed a hydrogen bonding-meditated hydrogel made from acrylic acid-N-acryloyl glycinamide (AA-NAGA) copolymers and tannic acids (TA), which formed repeatable specific adhesion to polar surfaces in an ultra-fast and robust way, but hardly adhered to nonpolar materials. It demonstrated up to five-fold increase in shear adhesive strength and interfacial adhesive toughness with external loading rates varying from 5 to 500 mm min-1. With a mechanochemical coupling model based on Monte Carlo simulations, we quantitatively revealed the nonlinear dependence of rate-sensitive interfacial adhesion on external loading, which was in good agreement with the experimental data. Likewise, the developed hydrogels were biocompatible, possessed antioxidant and antibacterial properties and promoted wound healing. This work not only reports a stimuli-responsive hydrogel adhesive suitable for multiple biomedical applications, but also offers an innovative strategy for bionic designs of smart hydrogels with loading rate-sensitive specific adhesion for various emerging areas including flexible electronics and soft robotics.

Purse-string contraction guides mechanical gradient-dictated heterogeneous migration of epithelial monolayer.

Mechanical heterogeneity has been recognized as an important role in mediating collective cell migration, yet the related mechanism has not been elucidated. Herein, we fabricate heterogeneous stiffness gradients by leveraging microelastically-patterned hydrogels with varying periodic distance. We observe that a decrease in the periodic distance of the mechanical heterogeneity is accompanied by an overall increase in the velocity and directionality of the migrating monolayer. Moreover, inhibition of ROCK- and myosin ⅡA- but not Rac1-mediated contraction reduces monolayer migration on the mechanically heterogeneous substrates. Furthermore, we find that F-actin and myosin ⅡA form purse-string at the leading edge on the mechanically heterogeneous substrates. Together, these findings not only show that the orientational cell-cell contraction promotes collective cell migration under the mechanical heterogeneity, but also demonstrate that the mechanosensation arising from large-scale cell-cell interactions through purse-string formation mediated cell-cell orientational contraction can feed back to regulate the reorganization of epithelial tissues. STATEMENT OF SIGNIFICANCE: By detecting the links between heterogenous rigidity and collective cell migration behavior at the molecular level, we reveal that collective cell migration in the mechanical heterogeneity is driven by ROCK- and myosin-ⅡA-dependent cytoskeletal tension. We confirm that cytoskeletal tension across the epithelial tissue is holistically linked through F-actin and myosin-ⅡA, which cooperate to form purse-string structures for modulating collective tissue behavior on the exogenous matrix with mechanical heterogeneity. Mechanical heterogeneity initiates tissue growth, remodelling, and morphogenesis by orientating cell contractility. Therefore, tensional homeostasis across large-scale cell interactions appears to be necessary and sufficient to trigger collective tissue behavior. Overall, these findings shed light on the role of mechanical heterogeneity in tissue microenvironment for reorganization and morphogenesis.

High-Throughput Electromechanical Coupling Chip Systems for Real-Time 3D Invasion/Migration Assay of Cells.

Cell invasion/migration through three-dimensional (3D) tissues is not only essential for physiological/pathological processes, but a hallmark of cancer malignancy. However, how to quantify spatiotemporal dynamics of 3D cell migration/invasion is challenging. Here, this work reports a 3D cell invasion/migration assay (3D-CIMA) based on electromechanical coupling chip systems, which can monitor spatiotemporal dynamics of 3D cell invasion/migration in a real-time, label-free, nondestructive, and high-throughput way. In combination with 3D topological networks and complex impedance detection technology, this work shows that 3D-CIMA can quantitively characterize collective invasion/migration dynamics of cancer cells in 3D extracellular matrix (ECM) with controllable biophysical/biomechanical properties. More importantly, this work further reveals that it has the capability to not only carry out quantitative evaluation of anti-tumor drugs in 3D microenvironments that minimize the impact of cell culture dimensions, but also grade clinical cancer specimens. The proposed 3D-CIMA offers a new quantitative methodology for investigating cell interactions with 3D extracellular microenvironments, which has potential applications in various fields like mechanobiology, drug screening, and even precision medicine.

Geometric constraint-triggered collagen expression mediates bacterial-host adhesion.

Cells living in geometrically confined microenvironments are ubiquitous in various physiological processes, e.g., wound closure. However, it remains unclear whether and how spatially geometric constraints on host cells regulate bacteria-host interactions. Here, we reveal that interactions between bacteria and spatially constrained cell monolayers exhibit strong spatial heterogeneity, and that bacteria tend to adhere to these cells near the outer edges of confined monolayers. The bacterial adhesion force near the edges of the micropatterned monolayers is up to 75 nN, which is ~3 times higher than that at the centers, depending on the underlying substrate rigidities. Single-cell RNA sequencing experiments indicate that spatially heterogeneous expression of collagen IV with significant edge effects is responsible for the location-dependent bacterial adhesion. Finally, we show that collagen IV inhibitors can potentially be utilized as adjuvants to reduce bacterial adhesion and thus markedly enhance the efficacy of antibiotics, as demonstrated in animal experiments.

DNA hydrogels as selective biomaterials for specifically capturing DNA, protein and bacteria.

The ability to selectively capture biomacromolecules and other components from solution has many important applications in biotechnology. However, capturing targets from solution while minimizing interference with the sample solution is still challenging. Here, we describe the design and assembly of a group of DNA hydrogels consisting of long single-stranded DNA produced by rolling amplification reaction (RCA) and crosslinked by DNA duplexes. The developed DNA hydrogels can selectively capture and separate oligonucleotides, proteins and bacteria from solution in situ without complex separation processes. Since such DNA hydrogels can capture their targets in the solution independently, multiple DNA hydrogels that target different compounds can be employed to separate different compounds in the solution at the same time. The work not only expands the application of DNA hydrogels, but also paves the way for developing novel selective biomaterials. STATEMENT OF SIGNIFICANCE: Biomaterials capable of selectively capturing various components have great potential in the field of biotechnology. Here, we proposed a new class of hydrogel composed of crosslinked long DNA strands for selectively capturing DNA, protein and bacteria. Unlike traditional polymeric hydrogels that have small meshes and limit macromolecule diffusion owing to the short distance between two adjacent crosslinks, the described DNA hydrogel has a much larger distance between its crosslinks because of the sequence designability of DNA, which allows easy diffusion of biomacromolecules through its networks and greatly expand its specific surface area. Moreover, the developed DNA hydrogel can also easily combine different aptamers to target different components via the Watson-Crick base pairing without making significant changes in its original design.