383 research outputs found
New Nanomaterials and Luminescent Optical Sensors for Detection of Hydrogen Peroxide
Accurate methods that can continuously detect low concentrations of hydrogen peroxide (H2O2) have a huge application potential in biological, pharmaceutical, clinical and environmental analysis. Luminescent probes and nanomaterials are used for fabrication of sensors for H2O2 that can be applied for these purposes. In contrast to previous reviews focusing on the chemical design of molecular probes for H2O2, this mini-review highlights the latest luminescent nanoparticular materials and new luminescent optical sensors for H2O2 in terms of the nanomaterial composition and luminescent receptor used in the sensors. The nanomaterial section is subdivided into schemes based on gold nanoparticles, polymeric nanoparticles with embedded enzymes, probes showing aggregation-induced emission enhancement, quantum dots, lanthanide-based nanoparticles and carbon based nanomaterials, respectively. Moreover, the sensors are ordered according to the type of luminescent receptor used within the sensor membranes. Among them are lanthanide complexes, metal-ligand complexes, oxidic nanoparticles and organic dyes. Further, the optical sensors are confined to those that are capable to monitor the concentration of H2O2 in a sample over time or are reusable. Optical sensors responding to gaseous H2O2 are not covered. All nanomaterials and sensors are characterized with respect to the analytical reaction towards H2O2, limit of detection (LOD), analytical range, electrolyte, pH and response time/incubation time. Applications to real samples are given. Finally, we assess the suitability of the nanomaterials to be used in membrane-based sensors and discuss future trends and perspectives of these sensors in biomedical research
Critical review of polymer and hydrogel deposition methods for optical and electrochemical bioanalytical sensors correlated to the sensor’s applicability in real samples
Sensors, ranging from in vivo through to single-use systems, employ protective membranes or hydrogels to enhance sample collection or serve as filters, to immobilize or entrap probes or receptors, or to stabilize and enhance a sensor’s lifetime. Furthermore, many applications demand specific requirements such as biocompatibility and non-fouling properties for in vivo applications, or fast and inexpensive mass production capabilities for single-use sensors. We critically evaluated how membrane materials and their deposition methods impact optical and electrochemical systems with special focus on analytical figures of merit and potential toward large-scale production. With some chosen examples, we highlight the fact that often a sensor’s performance relies heavily on the deposition method, even though other methods or materials could in fact improve the sensor. Over the course of the last 5 years, most sensing applications within healthcare diagnostics included glucose, lactate, uric acid, O2, H+ ions, and many specific metabolites and markers. In the case of food safety and environmental monitoring, the choice of analytes was much more comprehensive regarding a variety of natural and synthetic toxicants like bacteria, pesticides, or pollutants and other relevant substances. We conclude that more attention must be paid toward deposition techniques as these may in the end become a major hurdle in a sensor’s likelihood of moving from an academic lab into a real-world product
Influenza virus differentially activates mTORC1 and mTORC2 signaling to maximize late stage replication
<div><p>Influenza A virus usurps host signaling factors to regulate its replication. One example is mTOR, a cellular regulator of protein synthesis, growth and motility. While the role of mTORC1 in viral infection has been studied, the mechanisms that induce mTORC1 activation and the substrates regulated by mTORC1 during influenza virus infection have not been established. In addition, the role of mTORC2 during influenza virus infection remains unknown. Here we show that mTORC2 and PDPK1 differentially phosphorylate AKT upon influenza virus infection. PDPK1-mediated phoshorylation of AKT at a distinct site is required for mTORC1 activation by influenza virus. On the other hand, the viral NS1 protein promotes phosphorylation of AKT at a different site via mTORC2, which is an activity dispensable for mTORC1 stimulation but known to regulate apoptosis. Influenza virus HA protein and down-regulation of the mTORC1 inhibitor REDD1 by the virus M2 protein promote mTORC1 activity. Systematic phosphoproteomics analysis performed in cells lacking the mTORC2 component Rictor in the absence or presence of Torin, an inhibitor of both mTORC1 and mTORC2, revealed mTORC1-dependent substrates regulated during infection. Members of pathways that regulate mTORC1 or are regulated by mTORC1 were identified, including constituents of the translation machinery that once activated can promote translation. mTORC1 activation supports viral protein expression and replication. As mTORC1 activation is optimal midway through the virus life cycle, the observed effects on viral protein expression likely support the late stages of influenza virus replication when infected cells undergo significant stress.</p></div
On Immunologists and Microbiologists: Ground Zero in the Battle for Interdisciplinary Knowledge
The individual disciplines of microbiology and immunology are exploding with new information necessary for understanding host-pathogen relationships, infectious diseases, cancer, and autoimmunity. Because of overlapping scientific interests, immunologists and microbiologists often share common academic affiliations. The coexistence is uneasy. Significant problems arise because the groups have evolved different intellectual traditions. Pressures are intensified by sporadic changes in perceptions of their relative worth. As the mixing of microbiologists and immunologists can be likened to ground zero in the fight for interdisciplinary knowledge, it is useful, at this time of escalating data acquisition and growing appreciation for multidisciplinary research, to examine their histories, the challenges to amalgamation, and the advantages of their association for the advancement of knowledge and the delivery of protection against disease. The exploration supports a recommitment to integration of the disciplines and a proposal to facilitate this by inclusion of expertise bridging the areas
Enhanced Chemiluminescence of a Superior Luminol Derivative Provides Sensitive Smartphone‐Based Point‐of‐Care Testing with Enzymatic μPAD
Chemiluminescence (CL) provides ideal conditions for point-of-care testing (POCT) with wide dynamic ranges, superior sensitivities, and detection simplicity. It has not arrived routinely in the POCT field due to naturally low quantum yields of typical probes and the lack of sensitive low-cost detection devices. Here, we developed a universal microfluidic paper-based analytical device (μPAD) using l-lactate as model analyte. We demonstrate that a smartphone camera can compete with a scientific CCD camera as performance benchmark when using the strong CL emitter, m-carboxy luminol, resulting in extraordinary signal-to-noise ratios of 67. The μPAD provides CV<10 %, stability at room temperature for≥3 months and simple processing. Furthermore, the μPAD enables the detection of picomoles of the luminophore providing additional design flexibility. Thus, this new CL-μPAD is available for translating the many CL standard analytical assays performed in microtiter plates, microarrays or other more complex detection strategies to the POC
New terbium complex as a luminescent probe for determination of chlorogenic acid in green coffee and roasted coffee infusions
Green coffee is coming into vogue as a food that contains remarkable contents of antioxidants like chlorogenic acid (ChA) and induces mild stimulation to the consumer. While most methods for determination of ChA require chromatographic separation prior its quantitation, we present the first probe and a simple, sensitive and validated luminescence method for the determination of chlorogenic acid in green and roasted coffee infusion samples that does not require a chromatographic separation. ChA can remarkably quench the luminescence intensity of the Tb3+ complex with 1-(furan-2-ylmethyl)-4-hydroxy-N-(4-methylpyridin-2-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinoline-3-carboxamide (R3) in aqueous solution containing urotropine buffer at a near neutral pH 7.5 at λexc = 315 nm and λem = 545 nm. Under optimal conditions, the quenching of the luminescence intensity is directly proportional to the concentration of ChA in the range of 0.5–30 μg/mL, and the detection limit is 180 ng/mL. From measurements of luminescence decay time, it was determined that both static and dynamic quenching is induced upon coordination of ChA to Tb-R3. The related quenching constants are KS = 5.97∙104 M−1 and KD = 1.05⋅104 M−1. Finally, the method was applied successfully to the determination of ChA in real green and roasted coffee infusion samples and validated by HPLC to yield very closely matching concentrations of both methods. Therefore, this method can serve for a simple quality control of total ChA contents in coffee prior and after roasting
Optical pH Sensing in Milk: A Small Puzzle of Indicator Concentrations and the Best Detection Method
Optical chemical sensors can yield distinctively different responses that are dependent on the method applied for readout and evaluation. We therefore present a comprehensive study on the pH determined non-continuously with optical sensors in real milk samples by either photometry or colorimetry (via the RGB-readout of digital images) compared to the pH values obtained electrochemically by potentiometry. Additionally, the photometric determination of pH was conducted with single-wavelength and a dual wavelength ratiometric evaluation of the absorbance. It was found that both the precision and accuracy of the pH determined by photometry benefit from lower concentrations of bromocresol purple, which served as the pH indicator inside the sensor membrane. A further improvement is obtained by the ratiometric evaluation of the photometric sensor response. The pH values obtained from the colorimetric evaluation, however, gain in precision and accuracy if a higher concentration of the indicator is immobilized inside the sensor membrane. This has a major impact on the future fabrication of optical pH sensor membranes because they can be better tuned to match to the most precise and accurate range of the planned detection method
Cationic liposomes for generic signal amplification strategies in bioassays
Liposomes have been widely applied in bioanalytical assays. Most liposomes used bare negative charges to prevent non-specific binding and increase colloidal stability. Here, in contrast, highly stable, positively charged liposomes entrapping the fluorescent dye sulforhodamine B (SRB) were developed to serve as a secondary, non-specific label, and signal amplification tool in bioanalytical systems by exploiting their electrostatic interaction with negatively charged vesicles, surfaces, and microorganisms. The cationic liposomes were optimized for long-term stability (> 5 months) and high dye entrapment yield. Their capability as secondary, non-specific labels was first successfully proven through electrostatic interactions of cationic and anionic liposomes using dynamic light scattering, and then in a bioassay with fluorescence detection leading to an enhancement factor of 8.5 without any additional surface blocking steps. Moreover, the cationic liposomes bound efficiently to anionic magnetic beads were stable throughout magnetic separation procedures and could hence serve directly as labels in magnetic separation and purification strategies. Finally, the electrostatic interaction was exploited for the direct, simple, non-specific labeling of gram-negative bacteria. Isolated Escherichia coli cells were chosen as models and direct detection was demonstrated via fluorescent and chemiluminescent liposomes. Thus, these cationic liposomes can be used as generic labels for the development of ultrasensitive bioassays based on electrostatic interaction without the need for additional expensive recognition units like antibodies, where desired specificity is already afforded through other strategies
Highly sensitive interleukin 6 detection by employing commercially ready liposomes in an LFA format
Recent years have confirmed the ubiquitous applicability of lateral flow assays (LFA) in point-of-care testing (POCT). To make this technology available for low abundance analytes, strategies towards lower limits of detections (LOD), while maintaining the LFA’s ease of use, are still being sought. Here, we demonstrate how liposomes can significantly improve the LOD of traditional gold nanoparticle (AuNP)–based assays while fully supporting a ready-to-use system for commercial application. We fine-tuned liposomes towards photometric and fluorescence performance on the synthesis level and applied them in an established interleukin 6 (IL-6) immunoassay normally using commercial AuNP labels. IL-6’s low abundance (< 10 pg mL−1) and increasing relevance as prognostic marker for infections make it an ideal model analyte. It was found that liposomes with a high encapsulant load (150 mmol L−1 sulforhodamine B (SRB)) easily outperform AuNPs in photometric LFAs. Specifically, liposomes with 350 nm in diameter yield a lower LOD even in complex matrices such as human serum below the clinically relevant range (7 pg mL−1) beating AuNP by over an order of magnitude (81 pg mL−1). When dehydrated on the strip, liposomes maintained their signal performance for over a year even when stored at ambient temperature and indicate extraordinary stability of up to 8 years when stored as liquid. Whereas no LOD improvement was obtained by exploiting the liposomes’ fluorescence, an extraordinary gain in signal intensity was achieved upon lysis which is a promising feature for high-resolution and low-cost detection devices. Minimizing the procedural steps by inherently fluorescent liposomes, however, is not feasible. Finally, liposomes are ready for commercial applications as they are easy to mass-produce and can simply be substituted for the ubiquitously used AuNPs in the POCT market
Phages Enter the Fight against Colorectal Cancer
Intestinal microbiota undergo significant changes in colorectal cancer (CRC). Zheng et al. (Nat. Biomed. Eng., 2019) observe detrimental overpopulation of Fusobacterium nucleatum in mice and patients, suppressing the beneficial butyrate-producing Clostridium butyricum. Phage-guided irinotecan-loaded dextran nanoparticles promote release of bacterial-derived butyrate, while F. nucleatum and CRC cells are eliminated. These findings describe a possible novel therapeutic strategy for CRC. [Abstract copyright: Copyright © 2019 Elsevier Inc. All rights reserved.
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