Rapid, simultaneous detection of mycotoxins with smartphone recognition-based immune microspheres.

How to achieve simultaneous and rapid detection of various mycotoxins in food has important practical significance in the field of food processing and safety. In this paper, a smartphone immunoassay system based on hydrogel microspheres has been constructed to quickly detect two mycotoxins at the same time. The rapid detection system was reflected in the following three processes: (1) rapid separation of free matter after direct competition reaction based on hydrogel solid-phase carrier particles; (2) rapid detection process based on efficient catalytic function of enzymes; (3) fast capture and analysis of images based on smartphone software. Ochratoxin A (OTA) and zearalenone (ZEN) are secondary toxic metabolites of fungi that can contaminate a wide range of foods and feeds. OTA and ZEN were used as detection model molecules to verify the feasibility of the intelligent rapid detection system. The entire detection process was within 30 min, and the results were analyzed in only 10 s. Detection limits of mycotoxins OTA and ZEN are 0.7711 ng L-1 and 1.0391 ng L-1. The recoveries of both mycotoxins ranged from 76.72 to 122.05%. This study provides a universal rapid detection method for on-site application of large-scale food security testing. Schematic diagram of the construction of the smartphone detection system: The system is divided into three parts: detection, image capture and analysis, and result.

Background-free upconversion-encoded microspheres for mycotoxin detection based on a rapid visualization method.

Methods for detecting mycotoxins are very important because of the great health hazards of mycotoxins. However, there is a high background and low signal-to-noise ratio in real-time sensing, and therefore it is difficult to meet the fast, accurate, and convenient requirements for control of food quality. Here we constructed a quantitative fluorescence image analysis based on multicolor upconversion nanocrystal (UCN)-encoded microspheres for detection of ochratoxin A and zearalenone. The background-free encoding image signal of UCN-doped microspheres was captured by fluorescence microscopy under near-infrared excitation, whereas the detection image signal of phycoerythrin-labeled secondary antibodies conjugated to the microspheres was captured under blue light excitation. We custom-wrote an algorithm to analyze the two images for the same sample in 10 s, and only the gray value in the red channel of the secondary probe confirmed the quantity. The results showed that this novel detection platform performed feasible and reliable fluorescence image measurements by this method. Additionally, the limit of detection of was 0.34721 ng/mL for ochratoxin A and 0.41162 ng/mL for zearalenone. We envision that this UCN encoding strategy will be usefully applied for fast, accurate, and convenient testing of multiple food contaminants to ensure the safety of the food.

Smartphone recognition-based immune microparticles for rapid on-site visual data-sharing detection of Newcastle disease virus.

Since the last century, animal viruses have posed great threats to the health of humans and the farming industry. Therefore, virus control is of great urgency, and regular, timely, and accurate detection is essential to it. Here, we designed a rapid on-site visual data-sharing detection method for the Newcastle disease virus with smartphone recognition-based immune microparticles. The detection method we developed includes three major modules: preparation of virus detection vectors, sample detection, and smartphone image analysis with data upload. First, the hydrogel microparticles containing active carboxyl were manufactured, which coated nucleocapsid protein of NDV. Then, HRP enzyme-labeled anti-NP nanobody was used to compete with the NDV antibody in the serum for color reaction. Then the rough detection results were visible to the human eyes according to the different shades of color of the hydrogel microparticles. Next, the smartphone application was used to analyze the image to determine the accurate detection results according to the gray value of the hydrogel microparticles. Meanwhile, the result was automatically uploaded to the homemade cloud system. The total detection time was less than 50 min, even without trained personnel, which is shorter than conventional detection methods. According to experimental results, this detection method has high sensitivity and accuracy. And especially, it uploads the detection information via a cloud platform to realize data sharing, which plays an early warning function. We anticipate that this rapid on-site visual data-sharing detection method can promote the development of virus self-checking at home.

Recent advances in cellular optogenetics for photomedicine.

Since the successful introduction of exogenous photosensitive proteins, channelrhodopsin, to neurons, optogenetics has enabled substantial understanding of profound brain function by selectively manipulating neural circuits. In an optogenetic system, optical stimulation can be precisely delivered to brain tissue to achieve regulation of cellular electrical activity with unprecedented spatio-temporal resolution in living organisms. In recent years, the development of various optical actuators and novel light-delivery techniques has greatly expanded the scope of optogenetics, enabling the control of other signal pathways in non-neuronal cells for different biomedical applications, such as phototherapy and immunotherapy. This review focuses on the recent advances in optogenetic regulation of cellular activities for photomedicine. We discuss emerging optogenetic tools and light-delivery platforms, along with a survey of optogenetic execution in mammalian and microbial cells.

NIR-Responsive Photodynamic Nanosystem Combined with Antitumor Immune Optogenetics Bacteria for Precise Synergetic Therapy.

Photodynamic therapy (PDT) and immunotherapy are considered promising methods for the treatment of tumors. However, these treatment systems are still suffering from shortcomings such as hypoxia, easy metastasis, and delayed immune response during PDT. Therefore, it is still challenging to establish a programmed and rapid response immune combination therapy platform. Here, we construct a two-step synergetic therapy platform for the treatment of primary tumors and distant tumors using upconversion nanoparticles (UCNPs) and engineered bacteria as therapeutic media. In the first step, erbium ion (Er3+)-doped UCNPs act as a photoswitcher to activate the photosensitizer ZnPc to produce 1O2 for primary tumor therapy. In the second step, thulium ion (Tm3+)-doped UCNPs can emit blue-violet light under the excitation of near-infrared (NIR) light to activate the engineered bacteria to produce interferon (INF-γ) and release them in the intestine, which can not only treat tumors directly but also act with PDT to regulate immune pathways to activate the immune system, resulting in a joint immunotherapy effect to inhibit the growth of distant tumors. As a new type of programmatic combination therapy, we have proved that this platform can jointly activate the body's immune system during PDT and immunization treatment and can effectively inhibit tumor metastasis.

Hydrogel microcapsules containing engineered bacteria for sustained production and release of protein drugs.

Subcutaneous administration of sustained-release formulations is a common strategy for protein drugs, which avoids first pass effect and has high bioavailability. However, conventional sustained-release strategies can only load a limited amount of drug, leading to insufficient durability. Herein, we developed microcapsules based on engineered bacteria for sustained release of protein drugs. Engineered bacteria were carried in microcapsules for subcutaneous administration, with a production-lysis circuit for sustained protein production and release. Administrated in diabetic rats, engineered bacteria microcapsules was observed to smoothly release Exendin-4 for 2 weeks and reduce blood glucose. In another example, by releasing subunit vaccines with bacterial microcomponents as vehicles, engineered bacterial microcapsules activated specific immunity in mice and achieved tumor prevention. The engineered bacteria microcapsules have potential to durably release protein drugs and show versatility on the size of drugs. It might be a promising design strategy for long-acting in situ drug factory.

Light-Sensitive Lactococcus lactis for Microbe-Gut-Brain Axis Regulating via Upconversion Optogenetic Micro-Nano System.

The discovery of the gut-brain axis has proven that brain functions can be affected by the gut microbiota's metabolites, so there are significant opportunities to explore new tools to regulate gut microbiota and thus work on the brain functions. Meanwhile, engineered bacteria as oral live biotherapeutic agents to regulate the host's healthy homeostasis have attracted much attention in microbial therapy. However, whether this strategy is able to remotely regulate the host's brain function in vivo has not been investigated. Here, we engineered three blue-light-responsive probiotics as oral live biotherapeutic agents. They are spatiotemporally delivered and controlled by the upconversion optogenetic micro-nano system. This micro-nano system promotes the small intestine targeting and production of the exogenous L. lactis in the intestines, which realizes precise manipulation of brain functions including anxiety behavior, Parkinson's disease, and vagal afferent. The noninvasive and real-time probiotic intervention strategy makes the communiation from the gut to the host more controllable, which will enable the potential for engineered microbes accurately and effectively regulating a host's health.

NIR light-responsive bacteria with live bio-glue coatings for precise colonization in the gut.

Recombinant bacterial colonization plays an indispensable role in disease prevention, alleviation, and treatment. Successful application mainly depends on whether bacteria can efficiently spatiotemporally colonize the host gut. However, a primary limitation of existing methods is the lack of precise spatiotemporal regulation, resulting in uncontrolled methods that are less effective. Herein, we design upconversion microgels (UCMs) to convert near-infrared light (NIR) into blue light to activate recombinant light-responsive bacteria (Lresb) in vivo, where autocrine "functional cellular glues" made of adhesive proteins assist Lresb inefficiently colonizing the gut. The programmable engineering platform is further developed for the controlled and effective colonization of Escherichia coli Nissle 1917 (EcN) in the gut. The colonizing bacteria effectively alleviate DSS-induced colitis in mice. We anticipate that this approach could facilitate the clinical application of engineered microbial therapeutics to accurately and effectively regulate host health.

Optotheranostic Nanosystem with Phone Visual Diagnosis and Optogenetic Microbial Therapy for Ulcerative Colitis At-Home Care.

Ulcerative colitis (UC) is a relapsing disorder characterized by chronic inflammation of the intestinal tract. However, the home care of UC based on remote monitoring, due to the operational complexity and time-consuming procedure, restrain its widespread applications. Here we constructed an optotheranostic nanosystem for self-diagnosis and long-acting mitigations of UC at home. The system included two major modules: (i) A disease prescreening module mediated by smartphone optical sensing. (ii) Disease real-time intervention module mediated by an optogenetic engineered bacteria system. Recombinant Escherichia coli Nissle 1917 (EcN) secreted interleukin-10 (IL-10) could downregulate inflammatory cascades and matrix metalloproteinases; it is a candidate for use in the therapeutic intervention of UC. The results showed that the Detector was able to analyze, report, and share the detection results in less than 1 min, and the limit of detection was 15 ng·mL-1. Besides, the IL-10-secreting EcN treatment suppressed the intestinal inflammatory response in UC mice and protected the intestinal mucosa against injury. The optotheranostic nanosystems enabled solutions to diagnose and treat disease at home, which promotes a mobile health service development.

Author Correction: Upconversion optogenetic micro-nanosystem optically controls the secretion of light-responsive bacteria for systemic immunity regulation.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Upconversion optogenetic micro-nanosystem optically controls the secretion of light-responsive bacteria for systemic immunity regulation.

Chemical molecules specifically secreted into the blood and targeted tissues by intestinal microbiota can effectively affect the associated functions of the intestine especially immunity, representing a new strategy for immune-related diseases. However, proper ways of regulating the secretion metabolism of specific strains still remain to be established. In this article, an upconversion optogenetic micro-nanosystem was constructed to effectively regulate the specific secretion of engineered bacteria. The system included two major modules: (i) Modification of secretory light-responsive engineered bacteria. (ii) Optical sensing mediated by upconversion optogenetic micro-nanosystem. This system could regulate the efficient secretion of immune factors by engineered bacteria through optical manipulation. Inflammatory bowel disease and subcutaneously transplanted tumors were selected to verify the effectiveness of the system. Our results showed that the endogenous factor TGF-ß1 could be controllably secreted to suppress the intestinal inflammatory response. Additionally, regulatory secretion of IFN-γ was promoted to slow the progression of B16F10 tumor.

An efficient delivery of photosensitizers and hypoxic prodrugs for a tumor combination therapy by membrane camouflage nanoparticles.

Photodynamic therapy (PDT) is an oxygen-dependent, non-invasive cancer treatment. The hypoxia in the tumor environment limits the therapeutic effects of PDT. The combined delivery of photosensitizers and hypoxic prodrugs is expected to improve the efficacy of tumor treatment. In this paper, an erythrocyte and tumor cell membrane camouflage nanocarrier co-loaded with a photosensitizer (indocyanine green) and a hypoxic prodrug (tirapazamine) were used to combine PDT with chemotherapy. The system achieved less macrophage clearance through erythrocyte membranes and tumor-targeted tumor cell membranes, thereby inducing cell death and increasing tumor environment hypoxia by NIR irradiation of photosensitizers. Furthermore, the hypoxic environment activated TPZ to kill more tumor cells. In vivo results showed that the tumor inhibition rate of the drug-loaded nanoparticles increased from 34% to 64% after membrane modification. Moreover, the tumor inhibition rate of the photodynamic treatment group alone was only 47%, and the tumor inhibition rate after the combination was 1.3 times that of photodynamic therapy alone. Our platform is expected to contribute to the further application of cancer combination therapy.

A smartphone-based quantitative detection platform of mycotoxins based on multiple-color upconversion nanoparticles.

The detection of mycotoxins in food is urgently needed because they pose a significant threat to public health. In this study, we developed a quantitative detection platform for mycotoxins by integrating multicolor upconversion nanoparticle barcode technology with fluorescence image processing using a smartphone-based portable device. The multi-colored upconversion nanoparticle encoded microspheres (UCNMs) were used as encoded signals for detecting different mycotoxins simultaneously. After indirect competitive immunoassays using UCNMs, images could be captured by the portable device and the camera of a smartphone. Then, a self-written Android application, which is an HSV-based image recognition program installed on a smartphone, analyzed images and offered a reliable and accurate result in less than 1 min. The quantitative detection platform of mycotoxins proved to be feasible and reliable, and the limit of detection (LOD) was 1 ng, which was lower than that obtained from standard assays. This study demonstrates a method for detecting mycotoxins in food and other point of care analysis.