Visualization of Engineered M13 Phages Bound to Bacterial Targets by Transmission Electron Microscopy.

The filamentous phage M13 is one of the most well-studied and characterized phages, particularly since it was introduced as a scaffold for phage display, a technique to express and evolve fusion proteins on the M13 phage's coat to study protein or peptide binding interactions. Since phages can be engineered or evolved to specifically bind to a variety of targets, engineered M13 phages have been explored for applications such as drug delivery, biosensing, and cancer therapy, among others. Specifically, with the rising challenge of antimicrobial resistance among bacteria, chimeric M13 phages have been explored both as detection and therapeutic agents due to the flexibility in tuning target specificity. Transmission electron microscopy (TEM) is a powerful tool enabling researchers to directly visualize and characterize binding of phages to bacterial surfaces. However, the filamentous phage structure poses a challenge for this technique, as the phages have similar morphology to bacterial structures such as pili. In order to differentiate between bacterial structures and the filamentous phages, here we describe a protocol to prepare TEM samples of engineered M13 phages bound to bacterial cells, in which the phage virions have been specifically labeled by decoration of the major capsid proteins with gold nanoparticles. This protocol enables clear visualization and unambiguous identification of attached filamentous phages within the context of bacterial cells expressing numerous pili.

NIR-Mediated Pyroelectric Catalysis for Sustained ROS/RNS Generation and Advanced Cancer Therapy In Vivo via Injectable Hydrogel.

Exogenous electrical stimulation has attracted considerable attention due to the advantages of microelectric induction and subsequent biological effects such as actin reorganization and reactive oxygen species (ROS) generation. Herein, an injectable hydrogel of BPR-ARG@Gel (BAG) with pyroelectric BPR nanoparticle loading and l-arginine (ARG) introduction was fabricated for advanced cancer therapy in vivo. Due to the photothermal effect, the holes and electrons in BPR nanoparticles were separated to produce an open-circuit voltage and consequently catalyze water H2O to generate toxic superoxide (•O2-) and hydroxyl radicals (•OH). These ROS substances further oxidize ARG to produce NO for synergistic tumor treatments. The mice experiments indicated that the employment of BAG hydrogel incorporation with a near-infrared laser downregulated the heat shock protein and recruited immune cells with 5-fold-enhanced expression of proinflammatory cytokines of interferon-γ. It was also noteworthy that the injectable hydrogel of BAG substantially induced the generation of reactive oxygen/nitrogen species (ROS/RNS) with reliable biosafety and strong tumor inhibition. Overall, these findings have provided potentially new inspirations and a feasible strategy to translate this multifunctional hydrogel toward tumor therapy in a pyroelectric stimulation manner.

Integrin αvß3-targeted self-assembled polypeptide nanomicelles for efficacious sonodynamic therapy against breast cancer.

Sonodynamic therapy (SDT) is an advanced non-invasive cancer treatment strategy with moderate tissue penetration, less invasiveness and a reliable curative effect. However, due to the low stability, potential bio-toxicity and lack of tumor targeting capability of most sonosensitizers, the vast clinical application of SDT has been challenging and limited. Therefore, it is desirable to develop a novel approach to implement sonosensitizers to SDT for cancer treatments. In this study, an amphiphilic polypeptide was designed to effectively encapsulate rose bengal (RB) as a model sonosensitizer to form peptido-nanomicelles (REPNs). The as-fabricated REPNs demonstrated satisfactory tumor targeting and fluorescence performances, which made them superb imaging tracers in vivo. In the meantime, they generated considerable amounts of reactive oxygen species (ROS) to promote tumor cell apoptosis under ultrasound irradiation and showed excellent anti-tumor performance without obvious side effects. These engineered nanomicelles in combination with medical ultrasound may be used to achieve integrin αvß3-targeted sonodynamic therapy against breast cancer, and it is also a promising non-invasive cancer treatment strategy for clinical translations.

Improved phase-to-height mapping method combine with device attitude.

Phase-to-height mapping is one of the important processes in three dimensional phase measurement profilometry. But, in traditional phase-to-height mapping method, the measurement accuracy is affected by device attitude, so it needs saving a large amount of mapping equations to achieve high-quality phase-to-height mapping. In order to improve that, this paper proposes an improved phase-to-height mapping method combine with device attitude. Firstly, we get the unwrapped phase of the target. Then, using generalized regression neural network is used to reduce the offset of phase information at the same height due to the randomness of device attitude. Last, the phase-to-height mapping is completed by substituting the unwrapped phase (the difference between having detected object and no detected object) of eliminate the offset into improved phase-to-height mapping method. Experimental results show that the proposed method could achieve high-quality phase-to-height mapping with less mapping equation and less memory space. Compared with the nonlinear phase-to-height mapping method (probabilistic neural network to eliminate phase offset), its accuracy is improved by 44.30%. Compared with the nonlinear phase-to-height mapping method (radial basis function neural network to eliminate phase offset), the accuracy is improved by 39.58%.

External stimuli-triggered photodynamic and sonodynamic therapies in combination with hybrid nanomicelles of ICG@PEP@HA: laser vs. ultrasound.

The concept of combining external medical stimuli with internal functional biomaterials to achieve cancer-oriented treatments is being emergingly developed. Optical and acoustical activations have shown particular promise as non-invasive regulation modalities in cancer treatment and intervention. It is always challenging to leverage the contributions of optical and acoustical stimuli and find appropriate biomaterials to optimally match them. Herein, a type of hybrid nanomicelle (ICG@PEP@HA) containing ICG as a photo/sonosensitizer, an amphiphilic peptide for membrane penetration and hyaluronic acid for cluster determinant 44 (CD44) targeting was fabricated. Triggered by the external stimuli of laser and US irradiation, their photo/sonothermal performance, in vitro reactive oxygen species (ROS) production capability and tumor-targeting efficiency have been systematically evaluated. It was interestingly found that the external stimulus of laser irradiation induced a greater quantity of ROS, which resulted in significant cell apoptosis and tumor growth inhibition in the presence of ICG@PEP@HA. The individual analyses and corresponding rationales have been investigated. Meanwhile, these hybrid nanomicelles were administered into MDA-MB-231 tumor-bearing nude mice for PDT and SDT therapies and their biocompatibility assessment, and a prevailing PDT efficacy and reliable bio-safety have been evidenced based on the hematological analysis and histochemical staining. In summary, this study has validated a novel pathway to utilize these hybrid nanomicelles for laser/US-triggered localized tumor treatment, and the treatment efficiency may be leveraged by different external stimuli sources. It is also expected to give rise to full accessibility to clinical translations for human cancer treatments by means of the as-reported laser/US-nanomicelle combination strategy.

Straw-Derived Activated Carbon Decorated with Ag3PO4 for Organic Pollutant Removal by a Circular Degradation Mechanism: Adsorption and Photocatalysis.

The escalating problem of water pollution has become an urgent concern, as it significantly undermines people's quality of life and overall public health. The increasing severity of water pollution represents a global challenge, with profound implications for human society. In this study, hydrothermal carbonization coupled with alkaline activation was utilized to repurpose barley straw into activated carbon (AC) as an absorbent. Silver phosphate (Ag3PO4) was synthesized as a potent photocatalyst. Subsequent ultrasound-assisted loading integrated the robust adsorptive capabilities of the AC with the advanced photocatalytic efficiency of silver phosphate, resulting in a superior composite material (AC/Ag3PO4) and implementing a novel "absorption-photocatalysis" active circular degradation strategy to remove hazardous organics in water. Comprehensive characterization assays confirmed the successful synthesis and incorporation of Ag3PO4 onto the AC scaffold. The composite with a Ag3PO4 concentration of 3 wt % exhibited a high methylene blue (MB) removal efficiency of 99.4% within 100 min. The reaction rate of this composite surpassed that of standalone AC by a factor of 2.89. Furthermore, cyclic regeneration studies via adsorption-desorption methodologies revealed the composite's resilience and sustained performance. The MB removal efficiency was maintained at 85.5% over five consecutive cycles, demonstrating the composite's remarkable stability. The integration of adsorptive and photocatalytic functionalities within a single system mitigates potential secondary pollution arising during the AC's desorption phase and enhances the organic contaminant removal efficiency. Moreover, the utilization of this integrated material reduces the quantity of chemicals and energy required for conventional adsorption water treatment techniques, as the material harnesses sunlight or alternative light sources to catalyze contaminant decomposition. This reduces the dependence on chemical treatment agents, contributing to resource conservation and alleviating environmental burdens. This pioneering approach offers a novel paradigm for addressing pollutant challenges in aqueous environments.