To address the challenge of drug accumulation and penetration at the tumor site(s), herein we describe a first-in-class nanocarrier containing 24 copies each of two bioactive peptides (BAPs) genetically fused in frame to the 24 N-termini of a human ferritin H-type construct, named THE-10. The two BAPs are specific for PD-L1 and integrin αVβ3/αVβ5 plus Neuropilin (iRGD) respectively, conferring immune checkpoint blockade and drug-internalization properties. In turn, the THE-10 backbone brings 48 BAPs contiguous for synergism, prolonged blood half-life, and release into the tumor microenvironment upon conditional cleavage of a metalloprotease-sensitive site. Predicted THE-10 multitasking activity was experimentally supported as follows. Size-exclusion chromatography and surface plasmon resonance demonstrated BAP cleavage/release and receptor binding (nanomolar KD). Live-cell/time-lapse imaging demonstrated 4-fold-increased internalization of naked therapeutic antibodies, mirrored by enhanced cytotoxicity of the corresponding Antibody-Drug Conjugate. Slight antitumor effects were observed in vivo by treating immune checkpoint-sensitive syngeneic mouse colorectal model with THE-10 alone. Drug boosting was instead considerable on colorectal and pancreatic tumor allografts when THE-10 was co-administered with both small and large chemotherapeutic agents, outperforming the original iRGD cyclic peptide. Thus, THE-10 may enhance target therapy, chemotherapy and immunotherapy altogether, e.g. it candidates as a multitasking, all-round, antineoplastic therapy booster.

• MeSH
• Ferritins * chemistry   genetics   pharmacology   MeSH
• Immunotherapy * MeSH
• Humans MeSH
• Mice MeSH
• Cell Line, Tumor MeSH
• Nanoparticles * chemistry   MeSH
• Drug Carriers * chemistry   MeSH
• Antineoplastic Agents pharmacology   chemistry   MeSH
• Recombinant Proteins chemistry   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

The PB2 subunit of the influenza virus polymerase complex is essential for viral replication, primarily through a mechanism known as cap-snatching. In this process, PB2 binds to the 5' cap structure of host pre-mRNAs, enabling the viral polymerase to hijack the host transcriptional machinery. This binding facilitates the cleavage and integration of the capped RNA fragment into viral mRNA, thereby promoting efficient viral replication. Inhibiting the PB2-cap interaction is therefore crucial, as it directly disrupts the viral replication cycle. Consequently, targeting PB2 with specific inhibitors is a promising strategy for antiviral drug development against influenza. However, there are currently no available methods for the high-throughput screening of potential inhibitors. The development of new inhibitor screening methods of potential PB2 binders is the focus of this study. In this study, we present two novel methods, DIANA and AlphaScreen, for screening influenza PB2 cap-binding inhibitors and evaluate their effectiveness compared to the established differential scanning fluorimetry (DSF) technique. Using a diverse set of substrates and compounds based on the previously described PB2 binder pimodivir, we thoroughly assessed the capabilities of these new methods. Our findings demonstrate that both DIANA and AlphaScreen are highly effective for PB2 inhibitor screening, offering distinct advantages over traditional techniques such as isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR). These advantages include improved scalability, reduced sample requirements, and the capacity for label-free detection. Notably, DIANA's ability to determine Ki values from a single-well measurement significantly enhances its practicality and efficiency in inhibitor screening. This research represents a significant step forward in the development of more efficient and scalable screening strategies, helping advance efforts in the discovery of antiviral drugs against influenza.

• MeSH
• Antiviral Agents * pharmacology   chemistry   MeSH
• Fluorometry methods   MeSH
• Humans MeSH
• Piperidines pharmacology   MeSH
• Pyridines MeSH
• Pyrimidines MeSH
• Pyrroles MeSH
• RNA Caps metabolism   MeSH
• RNA-Dependent RNA Polymerase antagonists & inhibitors   metabolism   MeSH
• High-Throughput Screening Assays * methods   MeSH
• Viral Proteins * antagonists & inhibitors   metabolism   MeSH
• Influenza A virus drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

MXenes and their related nanocomposites with superior physicochemical properties such as high surface area, ease of synthesis and functionalization, high drug loading capacity, collective therapy potentials, pH-triggered drug release behavior, high photothermal conversion, and excellent photodynamic efficiency have been explored as alluring materials in photomedicine; the application of photons in medicine is facilitated for imaging and various disease treatment methods such as photothermal cancer/tumor ablation. Non-invasive theranostic strategies with synergistic activities have been developed using photothermal, photodynamic, and magnetic therapies together with remotely controlled drug/gene delivery for the diagnosis and treatment of various malignant diseases. Photothermal/photodynamic therapy and photoacoustic imaging using MXene-based structures have shown great promise in cancer phototherapy. However, hybridization and surface functionalization should be further explored to obtain biocompatible MXene-based composites/platforms with unique properties, high stability, and improved functionality in photomedicine. Toxicological and long-term biosafety assessments as well as clinical translation evaluations ought to be given high priority in research. Although some limited studies have revealed the excellent potentials of MXenes and their derivatives in photomedicine, further steps should be taken towards extensive research and detailed analysis in the field of optimizing the properties and improving the performance of these materials with a clinical and industrial outlook. Optical biosensing platforms have been developed along with electrochemical sensors and wearable sensors constructed from MXenes and their derivatives; future studies warrant the comprehensive analysis of optical transduction aspects such as colorimetry, electrochemiluminescence, photoluminescence, surface-enhanced Raman scattering, and surface plasmon resonance. Herein, the potentials of MXenes in photomedicine are deliberated encompassing important challenges and future research directions.

• MeSH
• Photochemotherapy * MeSH
• Phototherapy methods   MeSH
• Hyperthermia, Induced * methods   MeSH
• Humans MeSH
• Neoplasms * diagnostic imaging   drug therapy   MeSH
• Nanocomposites * chemistry   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

Microribonucleic acids (miRNAs) are short noncoding ribonucleic acids that have been linked with a multitude of human diseases including lung, breast, and hematological cancers. In this work, we present a novel, extremely sensitive assay for the label-free optical biosensor-based detection of miRNAs, which is based on the oligonucleotide-triggered release of nanoparticles from a sensor surface. We combine this assay (herein referred to as the nanoparticle-release (NPR) assay) with a surface plasmon resonance biosensor and show that the assay is able to enhance the specific sensor response associated with the binding of target miRNA while suppressing the interfering effects caused by the non-specific binding. We apply the assay to the detection of miRNAs related to myelodysplastic syndromes (miR-125b, miR-16) in blood plasma and demonstrate that the assay enables detection of miR-125b with a limit of detection (LOD) of 349 aM (corresponding to the lowest detectable amounts of 419 zmol). The achieved LOD is better by a factor of ∼100 when compared to the conventional nanoparticle-enhanced sandwich assay. Moreover, we demonstrate that the NPR assay may be combined with time-division multiplexing for the multiplexed miRNA detection.

• MeSH
• Biosensing Techniques * MeSH
• Metal Nanoparticles * MeSH
• Plasma MeSH
• Humans MeSH
• MicroRNAs * genetics   MeSH
• Myelodysplastic Syndromes * diagnosis   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Recently nanoparticle enhanced Laser Induced Breakdown Spectroscopy (NELIBS) is getting a growing interest as an effective alternative method for improving the analytical performance of LIBS. On the other hand, the plasmonic effect during laser ablation can be used for a different task rather than elemental analysis. In this paper, the dependence of NELIBS emission signal enhancement on nanoparticle-protein solutions dried on a reference substrate (metallic titanium) was investigated. Two proteins were studied: Human Serum Albumin (HSA) and Cytochrome C (CytC). Both proteins have a strong affinity for the gold nanoparticles (AuNPs) due to the bonding between the single free exterior thiol (associated with a cysteine residue) and the gold surface to form a stable protein corona. Then, since the protein sizes are vastly different, a different number of protein units is needed to cover AuNP surface to form a protein layer. The NP-protein solution was dropped and dried onto the titanium substrate. Then the NELIBS signal enhancement of Ti emission lines was correlated to the solution characteristics as determined with Dynamic Light Scattering (DLS), Surface Plasmon Resonance (SPR) spectroscopy and Laser Doppler Electrophoresis (LDE) for ζ-potential determination. Moreover, the dried solutions were studied with TEM (Transmission Electron Microscopy) for the inspection of the inter-particle distance. The structural effect of the NP-protein conjugates on the NELIBS signal reveals that NELIBS can be used to determine the number of protein units required to form the nanoparticle-protein corona with good accuracy. Although the investigated NP-protein systems are simple cases in biological applications, this work demonstrates, for the first time, a different use of NELIBS that is beyond elemental analysis and it opens the way for sensing the nanoparticle protein corona.

• MeSH
• Metal Nanoparticles * MeSH
• Lasers MeSH
• Humans MeSH
• Protein Corona * MeSH
• Spectrum Analysis MeSH
• Gold MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Autosomal dominant tubulointerstitial kidney disease (ADTKD)-uromodulin (UMOD) is the most common nonpolycystic genetic kidney disease, but it remains unrecognized due to its clinical heterogeneity and lack of screening test. Moreover, the fact that the clinical feature is a poor predictor of disease outcome further highlights the need for the development of mechanistic biomarkers in ADTKD. However, low abundant urinary proteins secreted by thick ascending limb cells, where UMOD is synthesized, have posed a challenge for the detection of biomarkers in ADTKD-UMOD. In the CRISPR/Cas9-generated murine model and patients with ADTKD-UMOD, we found that immunoglobulin heavy chain-binding protein (BiP), an endoplasmic reticulum chaperone, was exclusively upregulated by mutant UMOD in the thick ascending limb and easily detected by Western blot analysis in the urine at an early stage of disease. However, even the most sensitive ELISA failed to detect urinary BiP in affected individuals. We therefore developed an ultrasensitive, plasmon-enhanced fluorescence-linked immunosorbent assay (p-FLISA) to quantify urinary BiP concentration by harnessing the newly invented ultrabright fluorescent nanoconstruct, termed "plasmonic Fluor." p-FLISA demonstrated that urinary BiP excretion was significantly elevated in patients with ADTKD-UMOD compared with unaffected controls, which may have potential utility in risk stratification, disease activity monitoring, disease progression prediction, and guidance of endoplasmic reticulum-targeted therapies in ADTKD.NEW & NOTEWORTHY Autosomal dominant tubulointerstitial kidney disease (ADTKD)-uromodulin (UMOD) is an underdiagnosed cause of chronic kidney disease (CKD). Lack of ultrasensitive bioanalytical tools has hindered the discovery of low abundant urinary biomarkers in ADTKD. Here, we developed an ultrasensitive plasmon-enhanced fluorescence-linked immunosorbent assay (p-FLISA). p-FLISA demonstrated that secreted immunoglobulin heavy chain-binding protein is an early urinary endoplasmic reticulum stress biomarker in ADTKD-UMOD, which will be valuable in monitoring disease progression and the treatment response in ADTKD.

• MeSH
• Biomarkers urine   MeSH
• Immunosorbent Techniques * MeSH
• Nephritis, Interstitial genetics   urine   MeSH
• Humans MeSH
• Mice MeSH
• Heat-Shock Proteins urine   MeSH
• Endoplasmic Reticulum Stress physiology   MeSH
• Uromodulin genetics   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

A combined approach to signal enhancement in fluorescence affinity biosensors and assays is reported. It is based on the compaction of specifically captured target molecules at the sensor surface followed by optical probing with a tightly confined surface plasmon (SP) field. This concept is utilized by using a thermoresponsive hydrogel (HG) binding matrix that is prepared from a terpolymer derived from poly(N-isopropylacrylamide) (pNIPAAm) and attached to a metallic sensor surface. Epi-illumination fluorescence and SP-enhanced total internal reflection fluorescence readouts of affinity binding events are performed to spatially interrogate the fluorescent signal in the direction parallel and perpendicular to the sensor surface. The pNIPAAm-based HG binding matrix is arranged in arrays of sensing spots and employed for the specific detection of human IgG antibodies against the Epstein-Barr virus (EBV). The detection is performed in diluted human plasma or with isolated human IgG by using a set of peptide ligands mapping the epitope of the EBV nuclear antigen. Alkyne-terminated peptides were covalently coupled to the pNIPAAm-based HG carrying azide moieties. Importantly, using such low-molecular-weight ligands allowed preserving the thermoresponsive properties of the pNIPAAm-based architecture, which was not possible for amine coupling of regular antibodies that have a higher molecular weight.

• MeSH
• Acrylic Resins chemistry   MeSH
• Biosensing Techniques methods   MeSH
• Fluorescence MeSH
• Hydrogels chemistry   metabolism   MeSH
• Immunoglobulin G analysis   immunology   MeSH
• Epstein-Barr Virus Infections diagnosis   immunology   metabolism   virology   MeSH
• Humans MeSH
• Peptide Fragments immunology   metabolism   MeSH
• Polymers chemistry   MeSH
• Epstein-Barr Virus Nuclear Antigens immunology   MeSH
• Herpesvirus 4, Human immunology   isolation & purification   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Here, we present a new family of hierarchical porous hybrid materials as an innovative tool for ultrasensitive and selective sensing of enantiomeric drugs in complex biosamples via chiral surface-enhanced Raman spectroscopy (SERS). Hierarchical porous hybrid films were prepared by the combination of mesoporous plasmonic Au films and microporous homochiral metal-organic frameworks (HMOFs). The proposed hierarchical porous substrates enable extremely low limit of detection values (10-12 M) for pseudoephedrine in undiluted blood plasma due to dual enhancement mechanisms (physical enhancement by the mesoporous Au nanostructures and chemical enhancement by HMOF), chemical recognition by HMOF, and a discriminant function for bio-samples containing large biomolecules, such as blood components. We demonstrate the effect of each component (mesoporous Au and microporous AlaZnCl (HMOF)) on the analytical performance for sensing. The growth of AlaZnCl leads to an increase in the SERS signal (by around 17 times), while the use of mesoporous Au leads to an increase in the signal (by up to 40%). In the presence of a complex biomatrix (blood serum or plasma), the hybrid hierarchical porous substrate provides control over the transport of the molecules inside the pores and prevents blood protein infiltration, provoking competition with existing plasmonic materials at the limit of detection and enantioselectivity in the presence of a multicomponent biomatrix.

• MeSH
• Biosensing Techniques * MeSH
• Metal Nanoparticles * MeSH
• Plasma MeSH
• Pseudoephedrine MeSH
• Stereoisomerism MeSH
• Gold MeSH
• Publication type
• Journal Article MeSH

Our recent experience of the COVID-19 pandemic has highlighted the importance of easy-to-use, quick, cheap, sensitive and selective detection of virus pathogens for the efficient monitoring and treatment of virus diseases. Early detection of viruses provides essential information about possible efficient and targeted treatments, prolongs the therapeutic window and hence reduces morbidity. Graphene is a lightweight, chemically stable and conductive material that can be successfully utilized for the detection of various virus strains. The sensitivity and selectivity of graphene can be enhanced by its functionalization or combination with other materials. Introducing suitable functional groups and/or counterparts in the hybrid structure enables tuning of the optical and electrical properties, which is particularly attractive for rapid and easy-to-use virus detection. In this review, we cover all the different types of graphene-based sensors available for virus detection, including, e.g., photoluminescence and colorimetric sensors, and surface plasmon resonance biosensors. Various strategies of electrochemical detection of viruses based on, e.g., DNA hybridization or antigen-antibody interactions, are also discussed. We summarize the current state-of-the-art applications of graphene-based systems for sensing a variety of viruses, e.g., SARS-CoV-2, influenza, dengue fever, hepatitis C virus, HIV, rotavirus and Zika virus. General principles, mechanisms of action, advantages and drawbacks are presented to provide useful information for the further development and construction of advanced virus biosensors. We highlight that the unique and tunable physicochemical properties of graphene-based nanomaterials make them ideal candidates for engineering and miniaturization of biosensors.

• MeSH
• Betacoronavirus genetics   isolation & purification   pathogenicity   MeSH
• Biosensing Techniques * instrumentation   methods   trends   MeSH
• Equipment Design MeSH
• DNA, Viral analysis   genetics   MeSH
• Electrochemical Techniques MeSH
• Graphite * chemistry   MeSH
• Nucleic Acid Hybridization MeSH
• Clinical Laboratory Techniques * instrumentation   methods   statistics & numerical data   MeSH
• Colorimetry MeSH
• Coronavirus Infections diagnosis   epidemiology   virology   MeSH
• Quantum Dots chemistry   MeSH
• Humans MeSH
• Luminescence MeSH
• Nanostructures chemistry   MeSH
• Pandemics MeSH
• Surface Plasmon Resonance MeSH
• Spectrum Analysis, Raman MeSH
• Antigen-Antibody Reactions MeSH
• Virology methods   MeSH
• Pneumonia, Viral diagnosis   epidemiology   virology   MeSH
• Viruses genetics   isolation & purification   pathogenicity   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

The effects of combining naturally evolved photosynthetic pigment-protein complexes with inorganic functional materials, especially plasmonically active metallic nanostructures, have been a widely studied topic in the last few decades. Besides other applications, it seems to be reasonable using such hybrid systems for designing future biomimetic solar cells. In this paper, we describe selected results that point out to various aspects of the interactions between photosynthetic complexes and plasmonic excitations in Silver Island Films (SIFs). In addition to simple light-harvesting complexes, like peridinin-chlorophyll-protein (PCP) or the Fenna-Matthews-Olson (FMO) complex, we also discuss the properties of large, photosynthetic reaction centers (RCs) and Photosystem I (PSI)-both prokaryotic PSI core complexes and eukaryotic PSI supercomplexes with attached antenna clusters (PSI-LHCI)-deposited on SIF substrates.

• MeSH
• Chlorophyll A metabolism   MeSH
• Spectrometry, Fluorescence methods   MeSH
• Formaldehyde chemistry   MeSH
• Photosynthesis * MeSH
• Photosystem I Protein Complex metabolism   MeSH
• Glucose chemistry   MeSH
• Carotenoids metabolism   MeSH
• Nanostructures chemistry   ultrastructure   MeSH
• Silver chemistry   MeSH
• Light-Harvesting Protein Complexes metabolism   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH