Conventional diagnostic methods for biomarker detection often require invasive procedures and exhibit limited reproducibility and sensitivity. In this study, the turbulence-enhanced microneedle immunoassay platform (TMIP) was designed to enhance the performance and accuracy of biomarker detection in skin interstitial fluid (ISF). TMIP combines a bullet-shaped microneedle (MN) array for minimally invasive biomarker capture, a microfluidic device for MN-mediated immunoassay process simplification, and a star-shaped magnetic stirrer tool (MST) to facilitate efficient washing. By targeting S100 calcium-binding protein B (S100B), a diagnostic biomarker for melanoma, TMIP demonstrated substantial improvements in reproducibility, reducing signal deviations by up to 55 % compared to manual operation. The application of nanoporous MNs (NPMNs) achieved a low detection limit of 20 pg/mL with a high linearity (R2 = 0.9758). Validation using a gelatin phantom mimicking human skin confirmed TMIP's ability to achieve improved reproducibility and sensitivity. Furthermore, TMIP successfully detected S100B with high reproducibility in both the phantom (R2 = 0.97523) and melanoma-expressing mice within a rapid incubation time of 1 min. TMIP enables the detection of biomarkers with remarkable reproducibility and sub-nanogram sensitivity by simplifying the analysis process and enhancing reagent washing through turbulence. These features suggest that TMIP has the potential to serve as an efficient and reliable tool for biomarker detection in skin ISF.

Prostate cancer (PCa) is the second most commonly occurring cancer among men worldwide. Although the clinical management of PCa has significantly improved, a number of limitations have been identified in both early diagnosis and therapeutic treatment. Because multiple studies show that prostate-specific antigen (PSA) screening frequently results in overdiagnosis and overtreatment, the use of PSA alone as a diagnostic marker for PCa screening has been controversial. For individuals with locally advanced or metastatic PCa, androgen deprivation therapy (ADT) is the standard initially successful treatment; nonetheless, the majority of patients will eventually develop lethal metastatic castration-resistant prostate cancer (CRPC). Alternative treatment options, including chemo-, immuno-,or radio-therapy, can only prolong the survival of CRPC patients for several months with the most developing resistance. Considering this background, there is an urgent need to discuss about selective prostate-specific biomarkers that can predict clinically relevant PCa diagnosis and to develop biomarker-driven treatments to counteract CRPC. This review addresses several PCa-specific biomarkers that will assist physicians in determining which patients are at risk of having high-grade PCa, focusing on the clinical relevance of these biomarker-based tests among PCa patients. Secondly, this review highlights the effective use of these markers as drug targets to develop precision medicine or targeted therapies to counteract CRPC. Altogether, translating this biomarker-based research into the clinic will pave the way for the effective execution of personalized therapies for the benefit of healthcare providers, the biopharmaceutical industry, and patients.

This review provides a comprehensive overview of advanced analytical techniques employed in forensic science for paper comparison and differentiation, encompassing spectroscopic, chromatographic, mass spectrometric, and various complementary methods. It examines the principles, potential applications, and discriminatory potential of these diverse approaches, including the application of chemometrics and machine learning for data analysis and interpretation. However, the review critically highlights persistent challenges-such as substrate variability, environmental influences, database deficiencies, and validation gaps-which impede reliable forensic application. Enhancing scientific robustness and practical utility through focused efforts in validation, database creation, standardization, and interpretive methods is underscored as essential for advancing the field.

Although the most commonly used method for enhancing a limit of detection (LoD) in immunoassay is adopting chemiluminescence (CL), the liquid form of CL substrates has hindered its use for rapid diagnostic testing (RDT). In order to use the CL-based immunoassay in RDT with minimal user intervention, the liquid CL substrate should be converted to a dry form. In addition, a new RDT platform that is able to perform two sequential flows needs to be developed for the sequential flow control of the CL substrate. In this work, we have successfully developed a new dry form of CL substrate on the strip using a lyophilization process, as well as new lateral flow strips using an additional membrane pad for a time delay to achieve the desired sequential dual flows. Thus, on the dual-flow RDT strips, first the detection antibody conjugated with an enzyme flows over the test and control lines, and then the reconstituted CL substrate flows later. A hydrophilic PVDF membrane was selected as a pad material for the time delay to achieve the sequential dual flows through two flow paths, and flow introduction timing was functionally controlled to secure the time delay of approximately 5 minutes desired between the two flows. A CL-based cardiac troponin I (cTnI) assay was successfully performed on the new dual-flow RDT platform with a sample volume of 120 μL, achieving a LoD of 100 pg/mL. The achieved LoD is better than those possible with most of the currently available RDTs on the market. The new CL-based RDT platform with the capability of dual flows developed in this work can be used for numerous other immunodiagnostic platforms which need further high-sensitivity detection, envisaging a new RDT platform for point-of-care testing with further quantitative analysis.

A correlation between the emerging high case fatality rate of head and neck cancer and its propensity to migrate metastatically to other parts of the body makes it a significant global danger. It raises the demand for low-level detection, which is useful for early-stage diagnostics. Graphitic carbon nitride (g-C3N4) has recently garnered considerable attention as a promising nanomaterials for biosensor due to its exceptional redox behavior, electrochemical activity, and abundance of electroactive sites. The current study presents research outcomes regarding the development of an ultra-sensitive platform for detecting interleukin-8 (IL8), a cytokine associated with oral cancer. This investigation involves fabricating the platform using 3-aminopropyl trimethoxysilane (APTES)-functionalized g-C3N4 and assessing its efficacy in both laboratory-made and real samples. The process of g-C3N4 synthesis involved the thermal pyrolysis of urea without any add-on material. Moreover, the APTES@g-C3N4 nanomaterial was subjected to electrophoretic deposition onto an ITO-coated glass electrode. The fabricated APTES@g-C3N4/ITO electrode was covalently immobilized by the EDC and NHS chemical reaction in conjunction with anti-interleukin-8 (anti-IL8) antibodies. Before using these sensors for interleukin-8 (IL8) sensing, the anti-IL8/APTES@g-C3N4/ITO electrode was treated with bovine serum albumin (BSA) molecules utilized to obstruct non-targeted areas. Such a fabricated BSA/anti-IL8/APTES@g-C3N4/ITO electrochemical immunosensing bioelectrode was characterized by various analytical, morphological, and electrochemical techniques to confirm the stepwise fabrication of the sensor. BSA/anti-IL8/APTES@g-C3N4/ITO demonstrates a noticeable DPV based electrochemical response as a function of IL8 in the concentration ranging from 500 fg mL-1 to 160 ng mL-1. This BSA/anti-IL8/APTES@g-C3N4/ITO also exhibits a lower limit of detection (LOD) of 0.04 ng mL-1, a sensitivity of 0.015 mA log10 [ng mL-1] cm-2, and stability for up to 10 weeks. The biosensor demonstrates excellent performance in analyzing real samples, indicating its practical utility. This efficacy can be attributed to the abundance of electroactive sites, confined electronic structures, and strong interactions among the active g-C3N4 matrix, anti-IL8 molecules, and IL8 molecules. Our findings are essential for advancing early and point-of-care diagnostics, where quick turnaround times and great sensitivity are critical.

In this study, ionic-liquid submicron particles (ILSPs) encapsulating the luminophore tris(2',2-bipyridyl)ruthenium (II) ([Ru(bpy)3]2+) were developed as a carrier for an electrochemiluminescence immunoassay (ECLIA). The ILSPs were applied to quantitative determination of the model analyte prostate-specific antigen (PSA). The electrochemiluminescence of [Ru(bpy)3]2+ was measured with 2-(dibutylamino)ethanol as a co-reactant in nine ionic liquids (ILs). The electrochemiluminescence intensity was higher in 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide ([BMIM][TFSA]) and N-(2-methoxyethyl)-N-methyl pyrrolidinium bis(trifluoromethanesulfonyl)imide ([MEMP][TFSA]) than the other ILs. ILSPs were prepared using polyoxyethylene sorbitan monolaurate and sorbitan monooleate as surfactants and either [BMIM][TFSA] or [MEMP][TFSA]. The [BMIM][TFSA] ILSPs had a mean particle size of 244 nm and zeta potential of - 21.0 mV, and the [MEMP][TFSA] ILSPs had a mean particle size of 293 nm and zeta potential of - 17.9 mV. Microscope images showed that ILSPs were IL-in-water emulsions that completely encapsulated [Ru(bpy)3]2+. The ILSPs with [BMIM][TFSA] were more stable than those with [MEMP][TFSA], and [BMIM][TFSA] ILSPs was selected as a carrier in ECLIA for PSA determination. The calibration curve of PSA for ECLIA using the [BMIM][TFSA] ILSPs showed a good linear relationship (y = 0.29x + 4.02, r = 0.95) for the PSA concentration range of 100 pg/mL-100 μg/mL. The limit of detection and limit of quantification were 544 pg/mL and 35 ng/mL, respectively. Our results demonstrate that ECLIA using ILSPs can be used to easily determine the PSA concentration even with ILSPs in the particle state.

Adding pembrolizumab, an anti-PD-1 antibody approved for treatment of head and neck squamous cell carcinoma (HNSCC) to neoadjuvant (induction-) chemotherapy utilizing docetaxel and cisplatin (TP) followed by radiotherapy may improve outcome in larynx organ-preservation (LOP) that is investigated in the European Larynx-Organ preservation Study (ELOS). As biomarkers for response to TP and pembrolizumab +TP are missing but may include cytokines, this work aims on determining cytokines potentially linked to outcome as prognostic markers sufficient to predict and/or monitor response to successful LOP.

Collagenase IV digests were generated from 47 histopathological confirmed HNSCC tumor samples and seeded in 96-well plates containing pembrolizumab, docetaxel, cisplatin either solely or in binary or ternary combination. According to the FLAVINO protocol, supernatants were collected after 3 days, adherent cells fixed using ethanol, air-dried and pan-cytokeratin positive epithelial cells counted using fluorescence microscopy. The cytokines IL-6, IL-8, IFN-γ, IP-10, MCP-1, TNF-α, and VEGF in the supernatant were quantified by sandwich ELISA.

The mode of interaction between pembrolizumab and TP was assessed and correlated to outcome (overall, disease-specific and progression-free survival of patients). Suppression of MCP-1, IFN-γ and IL-6 production by pembrolizumab + TP exceeding the suppressive effect of TP was detected in the majority of samples and linked to improved survival. Multivariate Cox proportional hazard regression modeling revealed MCP-1, IFN-γ and IL-6 as independent outcome predictors.

Comparing response to TP vs. pembrolizumab vs. TP + pembrolizumab may allow for identification of patients with superior outcome independent from treatment applied.

In this study, multiple particle binding-liposomes (MPB-Lips), encapsulating the luminophore tris(2',2-bipyridyl)ruthenium (II) complex ([Ru(bpy)3]2+), were developed as an electrochemiluminescence (ECL) signal amplifier and were applied to detect the model analyte streptavidin (SA) using the indirect competitive ECL method. The MPB-Lips were prepared by mixing various ratios of two different liposomes-one containing a phospholipid with a primary amine group and a biotinyl group (BIO/NH2-Lip) and one containing a phospholipid with an N-hydroxysuccinimide group (NHS-Lip) to allow binding between particles via amide bonds. Quartz crystal microbalance analysis using SA-modified gold-coated quartz crystals showed that the frequency shift values of MPB-Lips gradually decreased in the order BIO/NH2-Lip:NHS-Lip = 1:0 < 1:1 < 1:3 < 1:5. This indicated that MPB-Lips were successfully formed. The indirect competitive ECL method using SA-modified gold electrodes showed that the 1:5-Lip system had greater sensitivity than the 1:0-Lip system-the limit of detection and quantification values for the systems were 1.84 and 6.30 μg mL-1 for 1:0-Lip, and 1.20 and 1.74 μg mL-1 for 1:5-Lip. Finally, the recovery of SA spiked in fetal bovine serum samples using the 1:5-Lip system showed good accuracy and precision with a recovery rate of 83-106% and relative standard deviation of 4-14%. Our study demonstrated that the MPB-Lips system was an effective and useful ECL amplifier and the ECL method using MPB-Lips could be applied to detect an analyte in a real sample.

The rise in antibiotic-resistant pathogens, highly infectious viruses, and chronic diseases has prompted the search for rapid and versatile medical tests that can be performed by the patient. Field-effect transistor (FET)-based electronic biosensing platforms are particularly attractive due to their sensitivity, fast turn-around time, potential for parallel detection of multiple pathogens, and compatibility with semiconductor manufacturing. However, an unmet critical need is a scalable, site-selective multiplexed biofunctionalization method with nanoscale precision for immobilizing different types of pathogen-specific bioreceptors on individual FETs, preventing parallel detection of multiple targets. Here, we propose a paradigm shift in FET biofunctionalization using thermal scanning probe lithography (tSPL) with a thermochemically sensitive polymer. This polymer can be spin-coated on fully-fabricated FET chips, making this approach applicable to any FET sensor material and technology. Crucially, we demonstrate the spatially selective multiplexed functionalization capability of this method by immobilizing different types of bioreceptors at prescribed locations on a chip with sub-20 nm resolution, paving the way for massively parallel FET detection of multiple pathogens. Antibody- and aptamer-modified graphene FET sensors are then realized, achieving ultra-sensitive detection of a minimum measured concentrations of 3 aM of SARS-CoV-2 spike proteins and 10 human SARS-CoV-2 infectious live virus particles per ml, and selectivity against human influenza A (H1N1) live virus.

This study explores the principles of resonance energy transfer and adsorption modulation using composites of Cu2S-MPA/NGODs. These composites can efficiently control the quenching process of electrochemiluminescence (ECL). Mercaptopropionic acid (MPA) was added during the synthesis of Cu2S-MPA to enhance its attachment to nitrogen-doped graphene quantum dots (NGODs). The UV absorption peaks of NGODs coincided with the emission peaks of luminol ECL, enabling resonance energy transfer and enhancing the quenching capability of Cu2S-MPA. Meanwhile, there is another quenching strategy. When the readily reducible Cu+ ions underwent partial reduction to Cu when they were bound to NGODs. This weakened the electrocatalytic effect on reactive oxygen species (ROS) and had a detrimental impact on electron transfer. Under optimal conditions, the immunosensor ECL intensity decreased linearly with the logarithm of carcinoembryonic antigen (CEA) concentration in the range of 0.00001-40 ng/mL, with a detection limit of 0.269 fg/mL. The sensor was effectively utilized for the identification of CEA in actual serum samples.

Current diagnostic tests for high sensitivity detection of protein biomarkers involve long incubation times or require bulky/expensive instrumentation, hindering their use for point-of-care testing. Here, we report a microfluidic electrochemical immunosensor that employs a unique finger-actuated mixer for rapid, ultrasensitive measurements of protein biomarkers. Mixing was implemented during the incubation steps, which accelerated biomolecular transport and promoted immunocomplex formation, leading to enhanced analytical sensitivity and a shortened detection time. Electrochemical measurements were performed using a handheld diagnostic device consisting of a smartphone and miniature potentiostat. Proof of principle was demonstrated by using this platform for quantitative measurements of C-X-C motif chemokine ligand 9 (CXCL9), a serological biomarker for autoimmune and inflammatory diseases, which could be detected in human plasma at concentrations as low as 4.7 pg mL-1 in <25 min. The ability to rapidly detect protein biomarkers with high sensitivity in a point-of-care format makes this device a promising tool for diagnostic testing, particularly in resource-limited settings.

The application of graphene-based nanocomposites for therapeutic and diagnostic reasons has advanced considerably in recent years due to advancements in the synthesis and design of graphene-based nanocomposites, giving rise to a new field of nano-cancer diagnosis and treatment. Nano-graphene is being utilized more often in the field of cancer therapy, where it is employed in conjunction with diagnostics and treatment to address the complex clinical obstacles and problems associated with this life-threatening illness. When compared to other nanomaterials, graphene derivatives stand out due to their remarkable structural, mechanical, electrical, optical, and thermal capabilities. The high specific surface area of these materials makes them useful as carriers in controlled release systems that respond to external stimuli; these compounds include drugs and biomolecules like nucleic acid sequences (DNA and RNA). Furthermore, the presence of distinctive sheet-like nanostructures and the capacity for photothermal conversion have rendered graphene-based nanocomposites highly favorable for optical therapeutic applications, including photothermal treatment (PTT), photodynamic therapy (PDT), and theranostics. This review highlights the current state and benefits of using graphene-based nanocomposites in cancer diagnosis and therapy and discusses the obstacles and prospects of their future development. Then we focus on graphene-based nanocomposites applications in cancer treatment, including smart drug delivery systems, PTT, and PDT. Lastly, the biocompatibility of graphene-based nanocomposites is also discussed to provide a unique overview of the topic.

In 2020, the World Health Organization (WHO) reported 604,000 new diagnoses of cervical cancer (CC) worldwide, and over 300,000 CC-related fatalities. The vast majority of CC cases are caused by persistent human papillomavirus (HPV) infections. HPV-related CC incidence and mortality rates have declined worldwide because of increased HPV vaccination and CC screening with the Papanicolaou test (PAP test). Despite these significant improvements, developing countries face difficulty implementing these programs, while developed nations are challenged with identifying HPV-independent cases. Molecular and proteomic information obtained from blood or tumor samples have a strong potential to provide information on malignancy progression and response to therapy in CC. There is a large amount of published biomarker data related to CC available but the extensive validation required by the FDA approval for clinical use is lacking. The ability of researchers to use the big data obtained from clinical studies and to draw meaningful relationships from these data are two obstacles that must be overcome for implementation into clinical practice. We report on identified multimarker panels of serum proteomic studies in CC for the past 5 years, the potential for modern computational biology efforts, and the utilization of nationwide biobanks to bridge the gap between multivariate protein signature development and the prediction of clinically relevant CC patient outcomes.

PCR amplification technology is the cornerstone of molecular biology. All-in-One PCR tube, as an emerging integrated device, is booming in biosensors application. All-in-One PCR tube biosensors are integrated PCR tubes designed for signal recognition, signal amplification or signal output. They enable "one-pot" detection within functionally modified and intelligently fabricated PCR tubes, effectively overcoming the limitations of conventional PCR applications, like complex procedural steps, risk of contamination and so on. Based on this, the review article summarizes the recent advance of All-in-One PCR tube biosensors for the first time as well as systematically categorizes five approaches of functional modification, three types of intelligent fabrication and relevant property characterization techniques. More emphasis is placed on the review of five ways of functional modification, including physical modification, chemical modification, UV photografting surface treatment, plasma surface modification, and layer-by-layer assembly coating. Moreover, All-in-One PCR tube biosensors covering different recognition elements range from small molecules to protein are detailed discussed on principle of sensing, providing a deeper understanding of the design and application of All-in-One-tube biosensor. Last, the future opportunities and challenges in this fascinating field are also deliberated.

Liver cancer is one of the deadliest types worldwide and early diagnosis is highly important for successful treatment. Therefore, it is necessary to develop rapid, sensitive, simple, and inexpensive analytical tools for its detection. MicroRNAs (miRNA) represent unique biomarkers whose expression in biofluids is strongly associated with cancer in general and miR-21, -31, -122, -145, -146a, -200c, -221, -222, and -223 in liver cancer, specifically. Various biosensors for miRNA detection have been developed. These include electrochemical biosensors based on amperometric, potentiometric, conductometric and impedimetric technology. Furthermore, the use of advanced nanomaterials with enhanced chemical stability, conductivity and electrocatalytic activity have greatly increased the sensitivity and specificity of these devices. The present review focuses on recent advances in electrochemical biosensors for miRNA detection in liver cancer.

This paper investigates an adaptive body biasing (ABB) circuit to improve the reliability and variability of a low-voltage inductor-capacitor (LC) voltage-controlled oscillator (VCO). The ABB circuit provides VCO resilience to process variability and reliability variation through the threshold voltage adjustment of VCO's transistors. Analytical equations considering the body bias effect are derived for the most important relations of the VCO and then the performance is verified using the post-layout simulation results. Under a 0.16% threshold voltage shift, the sensitivity of the normalized phase noise and transconductance of the VCO with the ABB circuit compared to the constant body bias (CBB) decreases by around 8.4 times and 3.1 times, respectively. Also, the sensitivity of the normalized phase noise and transconductance of the proposed VCO under 0.16% mobility variations decreases by around 1.5 times and 1.7 times compared to the CBB, respectively. The robustness of the VCO is also examined using process variation analysis through Monte Carlo and corner case simulations. The post-layout results in the 180 nm CMOS process indicate that the proposed VCO draws a power consumption of only 398 µW from a 0.6 V supply when the VCO frequency is 2.4 GHz. It achieves a phase noise of -123.19 dBc/Hz at a 1 MHz offset and provides a figure of merit (FoM) of -194.82 dBc/Hz.

Statistical analysis of the properties of single microparticles, such as cells, bacteria or plastic slivers, has attracted increasing interest in recent years. In this regard, field flow cytometry is considered the gold standard technique, but commercially available instruments are bulky, expensive, and not suitable for use in point-of-care (PoC) testing. Microfluidic flow cytometers, on the other hand, are small, cheap and can be used for on-site analyses. However, in order to detect small particles, they require complex geometries and the aid of external optical components. To overcome these limitations, here, we present an opto-fluidic flow cytometer with an integrated 3D in-plane spherical mirror for enhanced optical signal collection. As a result, the signal-to-noise ratio is increased by a factor of six, enabling the detection of particle sizes down to 1.5 µm. The proposed optofluidic detection scheme enables the simultaneous collection of particle fluorescence and scattering using a single optical fiber, which is crucial to easily distinguishing particle populations with different optical properties. The devices have been fully characterized using fluorescent polystyrene beads of different sizes. As a proof of concept for potential real-world applications, signals from fluorescent HEK cells and Escherichia coli bacteria were analyzed.

For the progress of point-of-care medicine, where individual health status can be easily and quickly monitored using a handheld sensor, saliva serves as one of the best-suited body fluids thanks to its availability and abundance of physiological indicators. Salivary biomarkers, combined with rapid and highly sensitive detection tools, may pave the way to new real-time health monitoring and personalized preventative therapy branches using saliva as a target matrix. Saliva is increasing in importance in liquid biopsy, a non-invasive approach that helps physicians diagnose and characterize specific diseases in patients. Here, we propose a proof-of-concept study combining the unique specificity in biomolecular recognition provided by surface-enhanced Raman spectroscopy (SERS) in combination with molecular dynamics (MD) simulations, which give leave to explore the biomolecular absorption mechanism on nanoparticle surfaces, in order to verify the traceability of two validated salivary indicators, i.e., interleukin-8 (IL-8) and lysozyme (LYZ), implicated in oropharyngeal squamous cell carcinoma (OSCC) and oral infection. This strategy simultaneously assures the detection and interpretation of protein biomarkers in saliva, ultimately opening a new route for the evolution of fast and accurate point-of-care SERS-based sensors of interest in precision medicine diagnostics.

As point-of-care testing (POCT) is becoming the new paradigm of medical diagnostics, there is a growing need to develop reliable POCT devices that can be conveniently operated in a minimally invasive manner. However, the clinical potential of POCT diagnostics is yet to be realized, mainly due to the limited and inconsistent amount of collected samples on these devices, undermining their accuracy. This study proposes a new biosensing platform modified with a functional polysuccinimide (PSI)-silica nanoparticle (SNP) composite system that can substantially increase the protein conjugation efficiency by modulating physicochemical interaction with proteins by several hundred percent from an unmodified device. The efficacy of this PSI-SNP system is further validated by applying it on the surface of a microneedle array (MN), which has emerged as a promising POCT device capable of accessing interstitial fluid through minimal penetration of the skin. This PSI-SNP MN is demonstrated to detect a wide array of proteins with high sensitivity on par with conventional whole serum analysis, validated by in vivo animal testing, effectively displaying broad applicability in biomedical engineering.

Immune-checkpoint blockade (ICB) of programmed-death-1 (PD-1) with pembrolizumab or nivolumab is approved for treating recurrent/metastatic (R/M) head and neck squamous cell carcinoma (HNSCC). NadiHN and ADRISK are phase IIB trials investigating in locally advanced (LA) HNSCC having low or high risk of recurrence the potential benefits from adding nivolumab to post-operative radiotherapy or pembrolizumab to cisplatin-based radio-chemotherapy.

Along five randomized controlled ICB trials including NadiHN and ADRISK, blood samples were taken before and after starting ICB in n=25 patients. Concentrations of vascular endothelial growth factor A (VEGF), CCL2 (MCP-1), interleukin-6 (IL-6), IL-8, interferon-gamma (IFN-γ), and CXCL10 (IP-10) pre- and post-ICB in EDTA-anticoagulated plasma and serum were compared. We used receiver operating characteristic (ROC) curves to identify optimal cutoff for defining subgroups before analyzing overall survival (OS) applying Kaplan-Meier plots and multivariate Cox regression.

We detected huge heterogeneity between cytokine patterns in pre-and post-ICB plasma and serum. We observed high correlation between concentrations of some cytokines. Despite absent systematic OS differences after ICB with pembrolizumab or nivolumab or between LA-HNSCC versus R/M HNSCC patients, we noticed improved outcome of patients having lower IFN-γ concentrations pre- and post-ICB and following ICB reduced concentrations of VEGF, IL-6, and IL-8 but not MCP-1. Contrarily, increases in IL-6, IL-8, and VEGF levels correlated with impaired outcome. Multivariate Cox regression revealed five independent OS predictors among cytokines; using natural logarithms of their hazard ratios to estimate an individual's risk of dying, three cytokine-expression pattern (CEP)-risk groups with no death within mean (95% confidence interval) follow-up of 29.2 (22.1-36.2) months and median OS of 11.3 (8.8-13.8) and 2.9 (0.4-5.4) months were found.

Whereas individual pre- or post-ICB cytokine concentrations in serum or plasma alone failed to predict the survivor group, CEP-risk groups may support the identification of individual patients with long-lasting benefit from ICB.

The sensitive analysis of glycoproteins is of great importance for early diagnosis and prognosis of diseases. In this work, a sandwich-type electrochemical aptasensor was developed for the detection of glycoproteins using 4-formylphenylboric acid (FPBA)-modified Cu-based metal-organic frameworks (FPBA-Cu-MOFs) as dual-functional signal probes. The target captured by the aptamer-modified electrode allowed the attachment of FPBA-Cu-MOFs based on the interaction between boronic acid and glycan on glycoproteins. Large numbers of Cu2+ ions in FPBA-Cu-MOFs produced an amplified signal for the direct voltammetric detection of glycoproteins. The electrochemical aptasensor showed a detection limit as low as 6.5 pg mL-1 for prostate specific antigen detection. The method obviates the use of antibody and enzymes for molecular recognition and signal output. The dual-functional MOFs can be extended to the design of other biosensors for the determination of diol-containing biomolecules in clinical diagnosis.

Rapid and reliable point-of-care (POC) diagnostic tests can have a significant impact on global health. One of the most common approaches for developing POC systems is the use of target-specific biomolecules. However, the conjugation of biomolecules can result in decreased activity, which may compromise the analytical performance and accuracy of the developed systems. To overcome this challenge, we present a polymer-based cross-linking protocol for controlled and directed conjugation of biological molecules. Our protocol utilizes a bifunctional thiol-polyethylene glycol (PEG)-hydrazide polymer to enable site-directed conjugation of IgG antibodies to the surface of screen-printed metal electrodes. The metal surface of the electrodes is first modified with thiolated PEG molecules, leaving the hydrazide groups available to react with the aldehyde group in the Fc fragments of the oxidized IgG antibodies. Using anti-Klebsiella pneumoniae carbapenemase-2 (KPC-2) antibody as a model antibody used for antimicrobial resistance (AMR) testing, our results demonstrate a ~10-fold increase in antibody coupling compared with the standard N-hydroxysuccinimide (NHS)-based conjugation chemistry and effective capture (>94%) of the target KPC-2 enzyme antigen on the surface of modified electrodes. This straightforward and easy-to-perform strategy of site-directed antibody conjugation can be engineered for coupling other protein- and non-protein-based biological molecules commonly used in POC testing and development, thus enhancing the potential for improved diagnostic accuracy and performance.

It is of great importance to have sensitive and accurate detection of cis-diol-containing biologically related substances because of their important functions in the research fields of metabolomics, glycomics, and proteomics. Boronic acids can specifically and reversibly interact with 1,2- or 1,3-diols to form five or six cyclic esters. Based on this unique property, boronic acid-based materials have been used as synthetic receptors for the specific recognition and detection of cis-diol-containing species. This review critically summarizes the recent advances with boronic acid-based materials as recognition elements and signal labels for the detection of cis-diol-containing biological species, including ribonucleic acids, glycans, glycoproteins, bacteria, exosomes, and tumor cells. We also address the challenges and future perspectives for developing versatile boronic acid-based materials with various promising applications.

Saliva is a versatile biofluid that contains a wide variety of biomarkers reflecting both physiologic and pathophysiologic states. Saliva collection is noninvasive and highly applicable for tests requiring serial sampling. Furthermore, advances in test accuracy, sensitivity and precision for saliva has improved diagnostic performance as well as the identification of novel markers especially in oral disease processes. These include dental caries, periodontitis, oral squamous cell carcinoma (OSCC) and Sjögren's syndrome (SS). Numerous growth factors, enzymes, interleukins and cytokines have been identified and are the subject of much research investigation. This review highlights current procedures for successful determination of saliva biomarkers including preanalytical factors associated with sampling, storage and pretreatment as well as subsequent analysis. Moreover, it provides an overview of the diagnostic applications of these salivary biomarkers in common oral diseases.

Due to their superior antibacterial properties, biocompatibility and high conductivity, nanomaterials have shown a broad prospect in the biomedical field and have been widely used in the prevention and treatment of oral diseases. Also due to their small particle sizes and biodegradability, nanomaterials can provide solutions for tissue engineering, especially for oral tissue rehabilitation and regeneration. At present, research on nanomaterials in the field of dentistry focuses on the biological effects of various types of nanomaterials on different oral diseases and tissue engineering applications. In the current review, we have summarized the biological effects of nanoparticles on oral diseases, their potential action mechanisms and influencing factors. We have focused on the opportunities and challenges to various nanomaterial therapy strategies, with specific emphasis on overcoming the challenges through the development of biocompatible and smart nanomaterials. This review will provide references for potential clinical applications of novel nanomaterials in the field of oral medicine for the prevention, diagnosis and treatment of oral diseases.

Protein mutation detection using liquid biopsy can be simply performed periodically, making it easy to detect the occurrence of newly emerging mutations rapidly. However, it has low diagnostic accuracy since there are more normal proteins than mutated proteins in body fluids. To increase the diagnostic accuracy, we analyzed plasma exosomes using nanoplasmonic spectra and deep learning. Exosomes, a promising biomarker, are abundant in plasma and stably carry intact proteins originating from mother cells. However, the mutated exosomal proteins cannot be detected sensitively because of the subtle changes in their structure. Therefore, we obtained Raman spectra that provide molecular information about structural changes in mutated proteins. To extract the unique features of the protein from complex Raman spectra, we developed a deep-learning classification algorithm with two deep-learning models. Consequently, controls with wild-type proteins and patients with mutated proteins were classified with high accuracy. As a proof of concept, we discriminated the lung cancer patients with mutations in the epidermal growth factor receptor (EGFR), L858R, E19del, L858R + T790M, and E19del + T790M, from controls with an accuracy of 0.93. Moreover, the protein mutation status of the patients with primary (E19del, L858R) and secondary (+T790M) mutations was clearly monitored. Overall, our technique is expected to be applied as a novel method for companion diagnostic and treatment monitoring.

Periostin is associated with several diseases, including cancers. Therefore, monitoring blood periostin levels is a powerful tool for diagnosing various diseases and identifying their severity. However, conventional detection methods pose several challenges, including high costs. To address these issues, we developed a novel one-shot dual aptamer-based fluorescence nanosensor for detecting periostin. The proposed nanosensor facilitates rapid, label-free, and sensitive detection of periostin using gold nanoprobes constructed by rhodamine-b isothiocyanate, PL2trunc aptamer, and gold nanoparticles and silver nanoprobes fabricated by the PL5trunc aptamer and silver nanoparticles. The two nanoprobes form a core-satellite structure by interacting with periostin, and the nanosensor detects periostin through the fluorescence regenerated by the increased proximity between them. The nanosensor successfully detected periostin with remarkable detection limits of 106.68 pM in buffer and 463.3 pM in serum-spiked conditions within 30 min without additional washing or signal amplification processes. Considering serum periostin levels in various diseases, the proposed nanosensor provides a suitable method for identifying patients with various diseases and determining disease severity. Moreover, the platform can be helpful as a practical method for on-site medical diagnosis because it can be adapted to detect other biomarkers simply by replacing the aptamer with other detection probes.

In this study, a novel split-type electrochemical immunosensor based on controlled release strategy was proposed for sensitive analysis and detection of tumor marker carbohydrate antigen 199 (CA19-9). Specifically, glucose (Glu) was encapsulated in carrier mesoporous silica (MSN) with encapsulation technology, and surface functionalized Zinc sulfide (ZnS) caps were used as "gatekeepers". The complex is formed by encapsulating Glu within MSN with ZnS (ZnS@MSN-Glu) as a signal amplifier labeled on the signal antibody (Ab₂). And the Ab₂ can detect the presence of antibodies. To reduce the interference of biological analysis, the immune recognition process of ZnS@MSN-Glu-Ab₂ bioconjugate and antigen was carried out in 96-well microplate, which did not interfere with the electrochemical analysis process. Therefore, the low sensitivity detection caused by biofouling of nanomaterials and immunoreaction on the testing platform is eliminated. Subsequently, the opening and timed release of mesopores were controlled by external stimuli, the disulfide bond cleavage by dithiothreitol (DTT), and glucose was effectively released. Then nickel cobalt layered double hydroxide (NiCo-LDH) were directly hydrothermally grown on carbon cloth (CC) electrodeposited with copper selenide (CuSe) nanosheets to construct three-dimensional (3D) cactus-like NiCo-LDH/CuSe/CC sensing platform. It can realize the catalytic oxidation of released glucose, triggering glucose-mediated signal amplification. The synergistic effect of the 3D cactus structure and active nanomaterials promotes electron conduction. Taking the detection of carbohydrate antigen CA19-9 as an example, the immunosensor shows a wide linear concentration range (0.001-100 U/mL) with the limit of detection of 0.0005 U/mL, realizing highly sensitive detection of CA19-9. This biosensing technique has considerable advantages and provides an innovative approach for trace detection of other biomarkers.

Photoelectrochemical (PEC) sensing is an emerging technological innovation for monitoring small substances/molecules in biological or non-biological systems. In particular, there has been a surge of interest in developing PEC devices for determining molecules of clinical significance. This is especially the case for molecules that are markers for serious and deadly medical conditions. The increased interest in PEC sensors to monitor such biomarkers can be attributed to the many apparent advantages of the PEC system, including an enhanced measurable signal, high potential for miniaturization, rapid testing, and low cost, amongst others. The growing number of published research reports on the subject calls for a comprehensive review of the various findings. This article is a review of studies on electrochemical (EC) and PEC sensors for ovarian cancer biomarkers in the last seven years (2016-2022). EC sensors were included because PEC is an improved EC; and a comparison of both systems has, expectedly, been carried out in many studies. Specific attention was given to the different markers of ovarian cancer and the EC/PEC sensing platforms developed for their detection/quantification. Relevant articles were sourced from the following databases: Scopus, PubMed Central, Web of Science, Science Direct, Academic Search Complete, EBSCO, CORE, Directory of open Access Journals (DOAJ), Public Library of Science (PLOS), BioMed Central (BMC), Semantic Scholar, Research Gate, SciELO, Wiley Online Library, Elsevier and SpringerLink.

In order to support biomolecule attachment, an effective electrochemical transducer matrix for biosensing devices needs to have many specialized properties, including quick electron transfer, stability, high surface area, biocompatibility, and the presence of particular functional groups. Enzyme-linked immunosorbent assays, gel electrophoresis, mass spectrometry, fluorescence spectroscopy, and surface-enhanced Raman spectroscopy are common techniques used to assess biomarkers. Even though these techniques provide precise and trustworthy results, they cannot replace clinical applications because of factors such as detection time, sample amount, sensitivity, equipment expense, and the need for highly skilled individuals. For the very sensitive and targeted electrochemical detection of the salivary oral cancer biomarker IL8, we have created a flower-structured molybdenum disulfide-decorated zinc oxide composite on GCE (interleu-kin-8). This immunosensor shows very fast detection; the limit of detection (LOD) for interleukin-8 (IL8) detection in a 0.1 M phosphate buffer solution (PBS) was discovered to be 11.6 fM, while the MoS₂/ZnO nanocomposite modified glassy carbon electrode (GCE) demonstrated a high catalytic current linearly from 500 pg to 4500 pg mL^-1 interleukin-8 (IL8). Therefore, the proposed biosensor exhibits excellent stability, high accuracy sensitivity, repeatability, and reproducibility and shows the acceptable fabrication of the electrochemical biosensors to detect the ACh in real sample analysis.

Sensors play a significant role in modern technologies and devices used in industries, hospitals, healthcare, nanotechnology, astronomy, and meteorology. Sensors based upon nanostructured materials have gained special attention due to their high sensitivity, precision accuracy, and feasibility. This review discusses the fabrication of graphene-based biosensors and gas sensors, which have highly efficient performance. Significant developments in the synthesis routes to fabricate graphene-based materials with improved structural and surface properties have boosted their utilization in sensing applications. The higher surface area, better conductivity, tunable structure, and atom-thick morphology of these hybrid materials have made them highly desirable for the fabrication of flexible and stable sensors. Many publications have reported various modification approaches to improve the selectivity of these materials. In the current work, a compact and informative review focusing on the most recent developments in graphene-based biosensors and gas sensors has been designed and delivered. The research community has provided a complete critical analysis of the most robust case studies from the latest fabrication routes to the most complex challenges. Some significant ideas and solutions have been proposed to overcome the limitations regarding the field of biosensors and hazardous gas sensors.

Cancer is the most challenging disease for medical professionals to treat. The factors underlying the complicated situation include anticancer drug-associated toxicity, non-specific response, low therapeutic window, variable treatment outcomes, development of drug resistance, treatment complications, and cancer recurrence. The remarkable advancement in biomedical sciences and genetics, over the past few decades, however, is changing the dire situation. The discovery of gene polymorphism, gene expression, biomarkers, particular molecular targets and pathways, and drug-metabolizing enzymes have paved the way for the development and provision of targeted and individualized anticancer treatment. Pharmacogenetics is the study of genetic factors having the potential to affect clinical responses and pharmacokinetic and pharmacodynamic behaviors of drugs. This chapter emphasizes pharmacogenetics of anticancer drugs and its applications in improving treatment outcomes, selectivity, toxicity of the drugs, and discovering and developing personalized anticancer drugs and genetic methods for prediction of drug response and toxicity.

Gene biomarkers of circulating tumor DNA (ctDNA) in liquid biopsies have been explored for use in the precise diagnosis of tumors. There is a great clinical need to realize the ultrasensitive detection of gene biomarkers in ctDNA. Here we reported that an ultrasensitive label-free biosensor was developed for the detection of the gastric cancer-related PIK3CA gene of ctDNA in peripheral blood. The polymeric l-arginine and graphene oxide-wrapped gold nanostars (rGO-AuNSs) were prepared and deposited on the glass electrode. The capturing DNA probes for the PIK3CA gene were prepared and successfully immobilized on the rGO-AuNS-modified electrode surface via π-π interaction among the rGO-AuNS composites and DNA probes. The resultant electrochemical sensor was effectively applied to detect the PIK3CA gene of ctDNA via the hybridization between the capturing DNA probe and ctDNA, the result of which showed that the biosensor exhibited desirable sensitivity, stability, and a wider dynamic response in a ctDNA concentration range from 1.0 × 10^-20 to 1.0 × 10^-10 M (R² = 0.997). Moreover, the low limit of detection of 1.0 × 10^-20 M (S/N = 3) indicates the biosensor owns satisfactory detection sensitivity. Fourteen PIK3CA genes and two PIK3CA gene mutations were detected in 60 clinical ctDNA samples of gastric cancer patients by using the developed biosensor. In conclusion, this ultrasensitive label-free electrochemical biosensor possesses a significant application prospect in the detection of the PIK3CA gene in ctDNA and in early screening for gastric cancer in the near future.

The early detection of biomarker proteins in clinical samples is of great significance for the diagnosis of diseases. However, it is still a challenge to detect low-concentration protein. Herein, a label-free aptamer-based amplification assay, termed the ATC-TA system, that allows fluorescence detection of very low numbers of protein without time-consuming washing steps and pre-treatment was developed. The target induces a conformational change in the allosteric aptasensor, triggers the target cycling and transcription amplification, and ultimately converts the input of the target protein into the output of the light-up aptamer (R-Pepper). It exhibits ultrahigh sensitivity with a detection limit of 5.62 fM at 37 ℃ and the accuracy is comparable to conventional ELISA. ATC-TA has potential application for the detection of endogenous PDGF-BB in serum samples to distinguish tumor mice from healthy mice at an early stage. It also successfully detects exogenous SARS-CoV-2 spike proteins in human serum. Therefore, this high-sensitive, universality, easy-to-operate and cost-effective biosensing platform holds great clinical application potential in early clinical diagnosis.

Monitoring the levels of cancer biomarkers is essential for cancer diagnosis and evaluation. In this study, a novel sandwich type sensing platform based on surface-enhanced Raman scattering (SERS) technology was developed for the detection of carcinoembryonic antigen (CEA), with a limit of detection (LOD) of 0.258 ng/mL. In order to achieve sensitive detection of CEA in complex samples, gold nanoparticle monolayer modified with CEA antibodies and with aptamer-functionalized probes was fabricated to target CEA. Two gold layers were integrated into the SERS platform, which greatly enhanced the signal of the probe by generating tremendous "hot spots". Meanwhile, the intensity ratio of Raman probes and the second-order peak of the silicon wafer was used to achieve dynamic calibration of the Raman probe signal. Excitingly, this sensing platform was capable of distinguishing cancer patients from healthy individuals via CEA concentrations in blood samples with the accuracy of 100%. This sandwich structure SERS sensing platform presented promising potential to be an alternative tool for clinical biomarker detection in the field of cancer diagnosis.

This study aims to assess whether preoperative platelet-lymphocyte ratio (PLR) and neutrophil-lymphocyte ratio (NLR) can predict occult metastasis in oral squamous cell carcinoma (OSCC).

Thirty-five OSCC cases were analyzed for clinicopathological and hematological data. Cases without metastasis (pN0) were checked for micrometastasis immunohistochemically using Desmoglein 3 (DSG3) and Cytokeratin (CK). Mean PLR and NLR were compared and analyzed between the study groups.

Metastatic deposits were detected in 9 out of 26 pN0 cases (34.6%) accounting for 11 out of 62 (17%) lymph nodes subjected to immunohistochemistry. The mean PLR was higher in OSCC cases with or without occult metastasis in comparison to controls (P < 0.001). No significant difference was found in the mean PLR and NLR between OSCC cases with and without occult metastasis. Furthermore, we found DSG3+ sinus histiocytes within the lymph nodes in majority of cases which is least reported in literature.

A significant percentage of cases showed occult metastasis in this study which led to upstaging of tumor. Although PLR was elevated in OSCC cases, it did not have a positive correlation with the presence of occult metastasis but was able to successfully distinguish OSCC patients from healthy individuals.

To improve the detection sensitivity of a porous silicon optical biosensor in the real-time detection of biomolecules, a non-spectral porous silicon optical biosensor technology, based on dual-signal light detection, is proposed. Double-light detection is a combination of refractive index change detection and fluorescence change detection. It uses quantum dots to label probe molecules to detect target molecules. In the double-signal-light detection method, the first detection-signal light is the detection light that is reflected from the surface of the porous silicon Bragg mirror. The wavelength of the detection light is the same as the wavelength of the photonic band gap edge of the porous silicon Bragg mirror. CdSe/ZnS quantum dots are used to label the probe DNA and hybridize it with the target DNA molecules in the pores of porous silicon to improve its effective refractive index and enhance the detection-reflection light. The second detection-signal light is fluorescence, which is generated by the quantum dots in the reactant that are excited by light of a certain wavelength. The Bragg mirror structure further enhances the fluorescence signal. A digital microscope is used to simultaneously receive the digital image of two kinds of signal light superimposed on the surface of porous silicon, and the corresponding algorithm is used to calculate the change in the average grey value before and after the hybridization reaction to calculate the concentration of the DNA molecules. The detection limit of the DNA molecules was 0.42 pM. This method can not only detect target DNA by hybridization, but also detect antigen by immune reaction or parallel biochip detection for a porous silicon biosensor.

Parathyroid hormone-related peptide (PTHrP) is related to bone metastasis and hypercalcemia in prostate and breast cancers and should be an excellent biomarker for aggressive forms of these cancers. Current clinical detection protocols for PTHrP are immunoradiometric assay and radioimmunoassay but are not sensitive enough to detect PTHrPs at early stages. We recently evaluated a prostate cancer biomarker panel, including serum monocyte differentiation antigen (CD-14), ETS-related gene protein, pigment epithelial-derived factor, and insulin-like growth factor-1, with promise for identifying aggressive prostate cancers. This panel predicted the need for patient biopsy better than PSA alone. In the present paper, we report an ultrasensitive microfluidic assay for PTHrPs and evaluate their diagnostic value and the value of including them with our prior biomarker panel to diagnose aggressive forms of prostate cancer. The immunoarray features screen-printed carbon sensor electrodes coated with 5 nm glutathione gold nanoparticles with capture antibodies attached. PTHrPs are bound to a secondary antibody attached to a polyhorseradish peroxidase label and delivered to the sensors to provide high sensitivity when activated by H₂O₂ and a mediator. We obtained an unprecedented 0.3 fg mL^-1 limit of detection for PTHrP bioactive moieties PTHrP 1-173 and PTHrP 1-86. We also report the first study of PTHrPs in a large serum pool to identify aggressive malignancies. In assays of 130 human patient serum samples, PTHrP levels distinguished between aggressive and indolent prostate cancers with 83-91% clinical sensitivity and 78-96% specificity. Logistic regression identified the best predictive model as a combination of PTHrP 1-86 and vascular endothelial growth factor-D. PTHrP 1-173 alone also showed a high ability to differentiate aggressive and indolent cancers.

Janus particles (JPs), named after the two-faced Roman god, are asymmetric particles with different chemical properties or polarities. JPs have been widely used in the biomedical field in recent years, including as drug carriers for targeted controlled drug release and as biosensors for biological imaging and biomarker detection, which is crucial in the early detection and treatment of diseases. In this review, we highlight the most recent advancements made with regard to Janus particles in point-of-care testing (POCT). Firstly, we introduce several commonly used methods for preparing Janus particles. Secondly, we present biomarker detection using JPs based on various detection methods to achieve the goal of POCT. Finally, we discuss the challenges and opportunities for developing Janus particles in POCT. This review will facilitate the development of POCT biosensing devices based on the unique properties of Janus particles.

Rapid and sensitive detection of cancer biomarkers is crucial for cancer screening, early detection, and improving patient survival rate. The present study proposes an electrochemical gene-sensor capable of detecting tumor related TP53 gene mutation hotspots by self-assembly of sulfhydryl ended hairpin DNA probes tagged with methylene blue (MB) onto a gold electrode. By performing a hybridization reaction with the target DNA sequence, the gene-sensor can rearrange the probe's structure, resulting in significant electrochemical signal differences by differential pulse voltammetry. When the DNA biosensor is hybridized with 1 μM target DNA, the peak current response signal can decrease more than 60%, displaying high sensitivity and specificity for the TP53 gene. The biosensor achieved rapid and sensitive detection of the TP53 gene with a detection limit of 10 nmol L-1, and showed good specific recognition ability for single nucleotide polymorphism (SNP) and base sequence mismatches in the TP53 gene affecting residue 248 of the P53 protein. Moreover, the biosensor demonstrated good reproducibility, repeatability, operational stability, and anti-interference ability for target DNA molecule in the complex system of 50% fetal bovine serum. The proposed biosensor provides a powerful tool for the sensitive and specific detection of TP53 gene mutation hotspot sequences and could be used in clinical samples for early diagnosis and detection of cancer.

Cytokines have been reported to alter the response to immune checkpoint inhibitors (ICIs) in patients with the tumor in accordance with their plasma concentrations. Here, we aimed to identify the key cytokines which influenced the responses and stimulated resistance to ICIs and tried to improve immunological response and develop novel clinical treatments in non-small cell lung cancer (NSCLC).

The promising predictive cytokines were analyzed via the multi-analyte flow assay. Next, we explored the correlation baseline level of plasma cytokines and clinical outcomes in 45 NSCLC patients treated with ICIs. The mechanism of the potential candidate cytokine in predicting response and inducing resistance to ICIs was then investigated.

We found NSCLC with a low baseline concentration of IL-6 in plasma specimens or tumor tissues could derive more benefit from ICIs based on the patient cohort. Further analyses revealed that a favorable relationship between PD-L1 and IL-6 expression was seen in NSCLC specimens. Results in vitro showed that PD-L1 expression in the tumor was enhanced by IL-6 via the JAK1/Stat3 pathway, which induced immune evasion. Notably, an adverse correlation was found between IL-6 levels and CD8⁺ T cells. And a positive association between IL-6 levels and myeloid-derived suppressor cells, M2 macrophages and regulator T cells was also seen in tumor samples, which may result in an inferior response to ICIs. Results of murine models of NSCLC suggested that the dual blockade of IL-6 and PD-L1 attenuated tumor growth. Further analyses detected that the inhibitor of IL-6 stimulated the infiltration of CD8⁺ T cells and yielded the inflammatory phenotype.

This study elucidated the role of baseline IL-6 levels in predicting the responses and promoting resistance to immunotherapy in patients with NSCLC. Our results indicated that the treatment targeting IL-6 may be beneficial for ICIs in NSCLC.

Protein biomarkers and microheterogeneity have attracted increasing attention in epidemiological and clinical research. Knowledge of within-person reproducibility over time is paramount to determine whether a single measurement accurately reflects an individual's long-term exposure. Yet, research investigating within-person reproducibility for proteoforms is limited. We investigated the reproducibility of the inflammatory markers C-reactive protein (CRP), serum amyloid A (SAA), and calprotectin (S100A8/9), and the renal function marker cystatin C (CnC) using a novel immuno-MALDI-TOF MS assay. Reproducibility, expressed as intraclass correlation coefficient (ICC), was calculated for 16 proteoforms using plasma samples of the Western Norway B Vitamin Intervention Trial (WENBIT) cohort collected 1-3 y apart from 295 stable angina pectoris (SAP) patients and 16 weeks apart from 38 subjects of the Intervention with Omega Fatty Acids in High-risk Patients with Hypertriglyceridemic Waist (OMEGA) trial with abdominal obesity but no other documented co-morbidities. ICCs for inflammatory markers were lower in WENBIT (CRP: 0.51, SAAt: 0.38, S100At: 0.31) compared to OMEGA subjects (CRP: 0.71, SAAt: 0.73, S100At: 0.48), while comparable for CnCt (WENBIT: 0.69, OMEGA: 0.67). Excluding SAP patients with elevated inflammation (CRP > 10 µg/ml) increased the ICC of SAAt to 0.55. Reduction of the time interval from 3 to 1 y in WENBIT group increased ICCs for all proteoforms. With a few exceptions ICCs did not differ between proteoforms of the same biomarker. ICCs were highest in OMEGA subjects with fair-to-good reproducibility for all markers. Reproducibility of SAA and S100A8/9 proteoforms in the WENBIT cohort was related to inflammation. This work will inform future clinical and epidemiological research which relies on single time point biomarker assessment to investigate inflammation and renal function.

With the increasing prevalence of diabetes in developing and developed countries, the socio-economic burden of diabetic retinopathy (DR), the leading complication of diabetes, is growing. Diabetic retinopathy (DR) is currently one of the leading causes of blindness in working-age adults worldwide. Robust methodologies exist to detect and monitor DR; however, these rely on specialist imaging techniques and qualified practitioners. This makes detecting and monitoring DR expensive and time-consuming, which is particularly problematic in developing countries where many patients will be remote and have little contact with specialist medical centres. Diabetic retinopathy (DR) is largely asymptomatic until late in the pathology. Therefore, early identification and stratification of vision-threatening DR (VTDR) is highly desirable and will ameliorate the global impact of this disease. A simple, reliable and more cost-effective test would greatly assist in decreasing the burden of DR around the world. Here, we evaluate and review data on circulating protein biomarkers, which have been verified in the context of DR. We also discuss the challenges and developments necessary to translate these promising data into clinically useful assays, to detect VTDR, and their potential integration into simple point-of-care testing devices.