The development of functional interfaces for healthcare applications is in great demand to perform sensitive and reliable testing. For this purpose, in the present study, the use of transition metal dichalcogenides (TMDCs) combined with carbon nanotubes modified pencil graphite electrode (PGE) was demonstrated for fish sperm double-stranded deoxyribonucleic acid (fsDNA) detection. The advanced system consisted of molybdenum disulfide (MoS2), tungsten disulfide (WS2), and multi-walled carbon nanotubes (MWCNTs), which exhibited superior and fascinating electrochemical properties on PGE as a result of synergetic effect occurred between the materials. The MoS2-WS2-MWCNTs incorporated PGE (MoS2-WS2-MWCNTs/E) was characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray diffraction (XRD) analysis, and electrochemical techniques. The elemental composition of the sensing platform was monitored with energy-dispersive X-ray spectroscopy (EDS). Specific parameters, including the amount of components in the hybrid material were optimized. MoS2-WS2-MWCNTs/E presented a good linear range for fsDNA between 0.1 and 50mgL-1 with a low detection limit of 0.05 mg L-1 by using the differential pulse voltammetry (DPV) technique. The detection performance of the resulting electrode was compared with the responses of MoS2-MWCNTs/E and WS2-MWCNTs/E as well. This sensing platform establishes the use of dichalcogenides-carbon nanotubes-based material for facile, sensitive, and reproducible sensing applications.

In this study, an innovative electrochemical biosensor was developed for the rapid, specific, and sensitive detection of Acinetobacter baumannii without the need for sample pretreatment. The biosensor utilized an aptamer as a specific capture probe for A. baumannii and employed a self-powered DNAzyme walker cleavage cycle reaction to achieve signal amplification. Upon introduction of the target bacteria, the aptamer captured the bacteria and released the Trigger, activating the DNAzyme to cleave the substrate chain containing methylene blue (MB). This led to the release of MB-labeled DNA fragments from the electrode surface, resulting in a significant decrease in the square wave voltammetry (SWV) signal of MB on the sensing platform. The limit of detection (LOD) for A. baumannii was determined to be 30 CFU/mL, enabling discrimination of the target bacteria from other common clinical isolates. Furthermore, the biosensor's potential for real sample analysis was demonstrated in cerebrospinal fluid (CSF), showcasing its efficacy and versatility as a biosensing tool with wide-ranging applications in disease diagnosis and bioanalysis.

Tetrahedral DNA frameworks (TDFs) have been extensively utilized in biosensing systems. At the macroscopic level, it has been well established that TDFs, when employed as probe scaffolds, significantly enhance the interfacial DNA hybridization efficiency between target single-stranded DNA (ssDNA) and probe ssDNA at the solid-liquid interface. However, a molecular-level understanding of how TDF scaffolds facilitate this interfacial hybridization remains elusive. In this study, we employed single-molecule total internal reflection fluorescence microscopy (SM-TIRFM) to monitor the hybridization processes between freely diffusing target ssDNA and probe ssDNA with or without TDF scaffolds. Statistical analysis of interfacial hybridization kinetics at single probe sites revealed that TDF scaffolds significantly accelerate the interfacial hybridization kinetics, reducing the target dissociation time by 0.5-fold and increasing the association constant (Ka) by nearly 4-fold. This study offers new insights for the development of DNA nanostructure-based biosensors.

The development of biosensors that can detect specific analytes continuously, in vivo, in real time has proven difficult due to biofouling, probe degradation and signal drift that often occur in vivo. By drawing inspiration from intestinal mucosa that can protect host cell receptors in the presence of the gut microbiome, we develop a synthetic biosensor that can continuously detect specific target molecules in vivo. The biomimetic multicomponent sensor features the hierarchical nano-bio interface design with three-dimensional bicontinuous nanoporous structure, polymer coating and aptamer switches, balancing small-molecule sensing and surface protection in complex biological environments. Our system is stable for at least 1 month in undiluted serum in vitro or 1 week implanted within the blood vessels of free-moving rats, retaining over 50% baseline signal and reproducible calibration curves. We demonstrate that the implanted system can intravenously track pharmacokinetics in real time even after 4 days of continuous exposure to flowing blood within rat femoral vein. In this way, our work provides a generalizable design foundation for biosensors that can continuously operate in vivo for extended durations.

Electrochemistry has become a key technique for studying biomolecular reactions and dynamics of living systems by using electron-transfer reactions to probe the complex interactions between biological redox molecules and their surrounding environments. To enable such measurements, redox mediators such as ferro/ferricyanide, ferrocene methanol, and tris(bipyridine) ruthenium(II) chloride are used. However, the impact of these exogeneous redox mediators on the health of cell cultures remains underexplored. Herein, we present the effects of three common redox mediators on the health of four of the most commonly used cell lines (Panc1, HeLa, U2OS, and MDA-MB-231) in biological studies. Cell health was assessed using three independent parameters: reactive oxygen species quantification by fluorescence flow cytometry, cell migration through scratch assays, and cell growth via luminescence assays. We show that as the concentration of mediator exceeds 1 mM, ROS increases in all cell types while cell viability plumets. In contrast, cell migration was only hindered at the highest concentration of each mediator. Our observations highlight the crucial role that optimized mediator concentrations play in ensuring accuracy when studying biological systems by electrochemical methods. As such, these findings provide a critical reference for selecting redox mediator concentrations for bioanalytical studies on live cells.

Exosomal RNA has emerged as a promising biomarker for glioblastoma (GBM) due to its exceptional stability in biofluids and strong correlation with tumor progression. In this study, we present an innovative intelligent molecular cleavage and dual-signal relay amplification-based ratiometric (ISR) strategy for high-sensitivity monitoring and dynamic analysis of exosomal RNA in glioma. The core mechanism is based on a hollow duplex structure that effectively prevents premature cleavage by duplex-specific nuclease (DSN), ensuring both the accuracy and stability of the detection system. Upon the introduction of the target microRNA (miRNA), one strand of the hollow duplex is displaced, forming a miRNA-DNA duplex that serves as a substrate for DSN, initiating target recycling and signal amplification. This dynamic process, coupled with dual-signal relay amplification, significantly enhances both sensitivity and stability, even at low miRNA concentrations. Our ratiometric approach substantially improves detection accuracy by comparing dual signal outputs. We further demonstrate the capability of real-time tracking of exosomal RNA dynamics, enabling precise monitoring of miRNA fluctuations over time. The practical applicability of our ISR strategy was validated by accurately detecting exosomal miRNA levels in clinical serum samples from glioblastoma patients, distinguishing them from healthy controls with high precision. Our method represents a significant advancement in early cancer detection and disease monitoring, with broad implications for precision medicine and the development of point-of-care diagnostic tools. Looking ahead, the ISR strategy holds great potential for monitoring a wide range of diseases, offering new opportunities for personalized diagnostics and therapeutic strategies.

Protein biosensors are significant tools in modern diagnostics due to their exceptional sensitivity and specificity in detecting protein biomarkers critical for disease diagnosis, therapeutic monitoring, and biomedical research. Innovations in transduction methods, nanomaterials, and point-of-care system integration have spurred recent advancements in biosensor technology. This summary examines key developments in protein biosensors, focusing on their structure, applications, and future potential. Nanomaterial-enhanced electrochemical biosensors, such as graphene, polyaniline, and carbon nanotubes, offer improved signal transmission due to their large surface area and faster electron transfer rates. Label-free immunosensors activated with gold nanoparticles and MXene-based sensors capable of combined biomarker analysis for detecting ovarian cancer are notable examples. During the COVID-19 pandemic, colorimetric and fluorescence optical biosensors facilitated easier diagnostics. An example of this is the incorporation of SARS-CoV-2 detection technologies into mobile phones. Real-time, label-free tracking with molecular precision is now possible due to the development of new methods, such as CRISPR-based platforms and quartz crystal microbalance (QCM)-based biosensors. This advancement is crucial for effectively managing infectious diseases and cancer. Synthetic fluorescence biosensors increase diagnostics by improving the visualization of protein interactions and cellular communication. Despite these achievements, challenges related to scalability, sustainability, and regulatory compliance remain. Proposed solutions include sustainable biosensor manufacturing, artificial intelligence-enhanced analytics for efficacy evaluation, and multidisciplinary approaches to optimize interaction with decentralised diagnostic systems. This work demonstrates how protein biosensors can advance precision medicine and global health.

Currently, mankind is fiercely struggling with cancer. Recently, we have been winning the battle against cancer through precision medicine and accompanying diagnostic methods, and we are raising many hopes with blockbuster drugs. It would be even better if we could read the cancer nucleotide sequence, identify them in advance, and suggest treatments simultaneously. However, this may be an impossible dream because it takes a lot of time and effort to diagnose and ensure all the long gene sequences of cancer at once. Thus, victory will be even closer if a rapid and accurate diagnosis of the cancer-specific gene biomarkers that will soon be imprinted can be made. With the advent of nanotechnology, a new short cancer diagnostic toolkit has been proposed to achieve the goal. This review presents a small diagnostic device that detects certain cancers' genetic fragments (simply 'Gizmo'). The development of numerous diagnostic methods has focused on (1) directly detecting pre-selectively targeted genes using novel diagnostic systems, and (2) indirectly detecting substantial improvements in diagnostic sensitivity only through detection signal amplification without existing gene amplification steps. Our fight against cancer is not a dream, but the result of success, and it is assumed that victory will accelerate as soon as possible.

Sustainable development represents a significant and pressing challenge confronting the global community at present. A wide variety of macroscopic engineering systems has been developed to promote sustainable development. Recent advancements in DNA materials have showcased their substantial contributions toward achieving sustainable development goals (SDGs). Compared to nonbiological materials, DNA materials possess exceptional properties such as genetic functionality, molecular programmability, recognition capabilities, and biocompatibility. These unique characteristics enable DNA materials to serve as general and versatile substrates beyond their genetic role. Consequently, they can be used to develop DNA-based engineering systems that offer versatile solutions to support sustainable development. In this Perspective, we critically examine the opportunities that DNA-based engineering systems present in contributing to the achievement of the SDGs within various real-world scenarios. We establish direct relationships between DNA-based engineering systems and the SDGs, highlighting their inherent merits in accelerating sustainable development. Furthermore, in order to successfully achieve SDGs, we address the challenges associated with these systems and emphasize the urgent need for developing multifunctional, reliable, biosafe, and intelligent DNA-based engineering systems to overcome these challenges.

This paper presents a low-cost, portable sensing platform for rapid DNA mass measurement, addressing a critical need in life science research. The platform features a novel interdigital open-gate junction field-effect transistor (ID-OGJFET) with a large sensing area that converts negatively charged DNA mass into an electrical current. The system enables DNA mass detection in under ten seconds with a resolution of less than 1 µA, demonstrating sensitivity across a range from 0.48 ng to 29.5 ng, achieving a Limit of Detection as low as 1.18-1.25 ng/µL. A custom-designed electronic reader and fluidic sample holder facilitate efficient operation. Simulation studies using molecular dynamics and finite element methods provide further insights into the sensor's DNA detection mechanism. This highly sensitive system is significantly more cost-effective than commercially available semiconductor characterization alternatives. The device's high performance and affordability make it a valuable tool for molecular biology applications, and it holds potential for advancing FET-based sensing instrumentation and measurement research.

The development of universal electrochemical sensing platforms with high sensitivity and specificity is of great significance for advancing practical disease diagnostic methods and devices. Exploring the structural properties of electrode materials and their interaction with biomolecules is essential to developing novel and distinctive analytical approaches. Here, we innovatively investigated the effect of DNA length and configuration on DNA molecule transfer into the nanostructure of a nanoporous gold (NPG) electrode. The NPG electrode can not only distinguish and quantify short DNA strands but can also prevent the diffusion of long DNA, thereby minimizing or eliminating background interference. Leveraging these findings, we developed a universal DNA-based NPG electrochemical biosensing platform for the detection of different types of biomolecules. As a proof-of-concept, this sensing platform was integrated with nuclease-assisted target-recycling recognition and amplification reactions to achieve sensitive and specific detection of single-stranded DNA, microRNA-21, and carcino-embryonic antigen, with detection limits of 4.09, 27.4, and 0.28 fM, respectively. The demonstrated universality, sensitivity, specificity, and capability for analyzing complex samples ensure a comprehensive and robust detection approach for nucleic acid-based molecular diagnosis.

Hydrogels are 3D networks of water-swollen hydrophilic polymers. It possesses unique properties (e.g., carrying biorecognition elements and creating a micro-environment) that make it highly suitable for bacteria detection (e.g., expedited and effective bacteria detection) and mitigation of bacterial contamination in specific environments (e.g., food systems). This study first introduces the materials used to create hydrogels for bacteria detection and the mechanisms for detection. We also summarize different hydrogel-based detection methods that rely on external stimuli and biorecognition elements, such as enzymes, temperature, pH, antibodies, and oligonucleotides. Subsequently, a range of widely utilized bacterial detection technologies were discussed where recently hydrogels are being used. These modifications allow for precise, real-time diagnostics across varied food matrices, responding effectively to industry needs for sensitivity, scalability, and portability. After highlighting the utilization of hydrogels and their role in these detection techniques, we outline limitations and advancements in the methods for the detection of foodborne pathogenic bacteria, especially the potential application of hydrogels in the food industry.

CD4 T lymphocytes play a key role in initiating the adaptive immune response, releasing cytokines that mediate numerous signal transduction pathways across the immune system. Therefore, CD4 T cell counts are widely used as an indicator of overall immunological health. HIV, one of the leading causes of death in the developing world, specifically targets and gradually depletes CD4 cells, making CD4 counts a critical metric for monitoring disease progression. As a result, accurately counting CD4 cells represents a pressing challenge in global healthcare. Flow cytometry remains the gold standard for enumerating CD4 T cells; however, flow cytometers are expensive, difficult to transport, and require skilled medical staff to prepare samples, operate the equipment, and interpret results. This highlights the critical need for novel, rapid, cost-effective, and portable methods of CD4 enumeration that are suitable for deployment in resource-limited countries. This review will survey and analyze emerging research in CD4 counting, with a focus on microfluidic systems, which represent a promising area of investigation.

In this study, we developed a simple pull-down assay using peptide nucleic acids (PNAs) equipped with a His-Tag and a G-quadruplex (G4) ligand for the selective recognition and quantification of G4-forming DNA sequences. Efficient and specific target recovery was achieved using optimized buffer conditions and magnetic Ni-NTA beads, while quantification was realized by employing the enzyme-like properties of the G4/hemin complex. The assay was validated through HPLC analysis and adapted for a 96-well plate format. The results show that higher recovery can be achieved using His-Tag with Ni-NTA magnetic beads as compared to the more common biotin-streptavidin purification. The inclusion of the G4-ligand as an additional selectivity handle was shown to be beneficial for both recovery and selectivity.

The COVID-19 epidemic and its continuous spread pose a serious threat to public health. Coronavirus strains known as SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) variants have undergone genomic changes. The severity of the symptoms, the efficiency of vaccinations, and the transmission capacity of the virus can be impacted by these alterations. Point-of-care diagnostic assays can identify particular genetic or protein sequences that are exclusive to each variety. Currently, ultrafast, responsive, and accurate antibody detection faces several challenges. Here, we outline the fabrication, implementation, and sensing performance benchmarking of an ultrafast (5 s) and inexpensive (0.15 USD) assay with label-free sensing of SARS-CoV-2 S (Spike)/N (Nucleocapsid) protein and other variants in real patient samples. A label-free DNA aptameric capacitive bio-sensing device was used to detect SARS-CoV-2 variants. Our novel, cutting-edge bio-sensing device contains a Wooden quoits conformation structural aptamer (WQCSA)-based inter-digitated capacitor electronic (WQCSA-IDCE) system. WQCSA-aptamer was used as a switch-turn on response to achieve ultrasensitivity in the variable area of the SARS-CoV-2. The molecular beacon (MB) method was also used to measure the fluorescently colored SARS-CoV-2 S/N protein. These sensors can be used with several types of label-free DNA aptamers to act as rapid, affordable, and label-free biosensors for a variety of critical acute respiratory virus syndrome disorders.

In this study, we propose a strategy to explore the impact of the proportion of tris(2-carboxyethyl)phosphine (TCEP) and 6-mercaptohexanol (MCH) on the efficiency of oligonucleotide functionalization on PDMS microfluidic channels equipped with pairs of homemade microfabricated platinum microelectrodes. We identified an optimal concentration of these compounds that enables the effective orientation and distribution of probes, thereby facilitating subsequent target hybridization. The experiment included optimizing sample injection into microfluidic channels. We used TCEP as a reducing agent to help the DNA probes adhere to the channel electrode better. This stopped the formation of disulfide bonds during the probe immobilization step. We found the optimal TCEP/MCH mixture ratio (5 mM TCEP and 50 mM MCH), which led to a more uniform distribution and orientation of the DNA probes on the platinum electrode. These optimized conditions resulted in a more compact DNA monolayer and enhanced detection capabilities. The biosensor's performance was evaluated by the detection of the hybridization of complementary DNA sequences in the presence of equimolar Fe(CN)63-/Fe(CN)64-. The detection of the synthetic GP8 resistance gene is facilitated by a measurable decrease in the electron transfer rate, which is directly proportional to its concentration. Under the optimized conditions, the DNA biosensor showed excellent sensitivity (with a detection limit of 10-17 M) and high specificity when tested against noncomplementary DNA strands.

Advances in protein engineering-enabled enzyme immobilization technologies have significantly improved enzyme-electrode wiring in enzymatic electrochemical systems, which harness natural biological machinery to either generate electricity or synthesize biochemicals. In this review, we provide guidelines for designing enzyme-electrodes, focusing on how performance variables change depending on electron transfer (ET) mechanisms. Recent advancements in enzyme immobilization technologies are summarized, highlighting their contributions to extending enzyme-electrode sustainability (up to months), enhancing biosensor sensitivity, improving biofuel cell performance, and setting a new benchmark for turnover frequency in bioelectrocatalysis. We also highlight state-of-the-art protein-engineering approaches that enhance enzyme-electrode interfacing through three key principles: protein-protein, protein-ligand, and protein-inorganic interactions. Finally, we discuss prospective avenues in strategic protein design for real-world applications.

Sensitive and quantitative detection of chemical and biological molecules for screening, diagnosis and monitoring diseases is essential to treatment planning and response monitoring. Electrochemical biosensors are fast, sensitive, and easy to miniaturize, which has led to rapid development in clinical diagnosis. Benefiting from their excellent molecular recognition ability and high programmability, DNA nanomaterials could overcome the Debye length of electrochemical biosensors by simple molecular design and are well suited as recognition elements for electrochemical biosensors. Therefore, to enhance the sensitivity and specificity of electrochemical biosensors, significant progress has been made in recent years by optimizing the DNA nanomaterials design. Here, the establishment of electrochemical sensing strategies based on DNA nanomaterials is reviewed in detail. First, the structural design of DNA nanomaterial is examined to enhance the sensitivity of electrochemical biosensors by improving recognition and overcoming Debye length. In addition, the strategies of electrical signal transduction and signal amplification based on DNA nanomaterials are reviewed, and the applications of DNA nanomaterial-based electrochemical biosensors and integrated devices in clinical diagnosis are further summarized. Finally, the main opportunities and challenges of DNA nanomaterial-based electrochemical biosensors in detecting disease biomarkers are presented in an aim to guide the design of DNA nanomaterial-based electrochemical devices with high sensitivity and specificity.

Research on self-assembled deoxyribonucleic acid (DNA) nanostructures with different shapes, sizes, and functions has recently made rapid progress owing to its biocompatibility, programmability, and stability. Among these, triangular unit-based DNA nanostructures, which are typically multi-arm DNA tiles, have been widely applied because of their unique structural rigidity, spatial flexibility, and cell permeability. Triangular unit-based DNA nanostructures are folded from multiple single-stranded DNA using the principle of complementary base pairing. Its shape and size can be determined using pre-set scaffold strands, segmented base complementary regions, and sequence lengths. The resulting DNA nanostructures retain the desired sequence length to serve as binding sites for other molecules and obtain satisfactory results in molecular recognition, spatial orientation, and target acquisition. Therefore, extensive research on triangular unit-based DNA nanostructures has shown that they can be used as powerful tools in the biosensing field to improve specificity, sensitivity, and accuracy. Over the past few decades, various design strategies and assembly techniques have been established to improve the stability, complexity, functionality, and practical applications of triangular unit-based DNA nanostructures in biosensing. In this review, we introduce the structural design strategies and principles of typical triangular unit-based DNA nanostructures, including triangular, tetrahedral, star, and net-shaped DNA. We then summarize the functional properties of triangular unit-based DNA nanostructures and their applications in biosensing. Finally, we critically discuss the existing challenges and future trends.

Self-assembled supramolecular DNA tetrahedra composed of programmed sequence-engineered complementary base-paired strands represent elusive nanostructures having key contributions to the development and diverse applications of DNA nanotechnology. By appropriate engineering of the strands, DNA tetrahedra of tuneable sizes and chemical functionalities were designed. Programmed functionalities for diverse applications were integrated into tetrahedra structures including sequence-specific recognition strands (aptamers), catalytic DNAzymes, nanoparticles, proteins, or fluorophore. The article presents a comprehensive review addressing methods to assemble and characterize the DNA tetrahedra nanostructures, and diverse applications of DNA tetrahedra framework are discussed. Topics being addressed include the application of structurally functionalized DNA tetrahedra nanostructure for the assembly of diverse optical or electrochemical sensing platforms and functionalized intracellular sensing and imaging modules. In addition, the triggered reconfiguration of DNA tetrahedra nanostructures and dynamic networks and circuits emulating biological transformations are introduced. Moreover, the functionalization of DNA tetrahedra frameworks with nanoparticles provides building units for the assembly of optical devices and for the programmed crystallization of nanoparticle superlattices. Finally, diverse applications of DNA tetrahedra in the field of nanomedicine are addressed. These include the DNA tetrahedra-assisted permeation of nanocarriers into cells for imaging, controlled drug release, active chemodynamic/photodynamic treatment of target tissues, and regenerative medicine.

Food processing unavoidably introduces various risky ingredients that threaten food safety. N-Nitrosamines (NAs) constitute a class of food contaminants, which are considered carcinogenic to humans. According to the compiled information, pretreatment methods based on solid-phase extraction (SPE) were widely used before the determination of volatile NAs in foods. The innovation of adsorbents and hybridization of other methods have been confirmed as the future trends of SPE-based pretreatment methods. Moreover, technologies based on liquid chromatography and gas chromatography were popularly applied for the detection of NAs. Recently, sensor-based methods have garnered increasing attention due to their efficiency and flexibility. More portable sensor-based technologies are recommended for on-site monitoring of NAs in the future. The application of artificial intelligence can facilitate data processing during high-throughput detection of NAs. Natural bioactive compounds have been confirmed to be effective in mitigating NAs in foods through antioxidation, scavenging precursors, and regulating microbial activities. Meanwhile, they exhibit strong protective activities against hepatic damage, pancreatic cancer, and other NA injuries. Further supplementation of data on the bioavailability of bioactives can be achieved through encapsulation and clinical trials. The utilization of bioinformatics tools rooted in various omics technologies is suggested for investigating novel mechanisms and finally broadening their applications in targeted therapies.

Understanding the growth of mesoporous crystalline materials, such as mesoporous metals, on different substrates can provide valuable insights into the crystal growth dynamics and the redox reactions that influence their electrochemical sensing performance. Herein, it is demonstrated how the amorphous nature of the glass substrate can suppress the typical <111> oriented growth in mesoporous Au (mAu) films. The suppressed <111> growth is manifested as an accumulation of strain, leading to the generation of abundant surface defects, which are beneficial for enhancing the electrochemical activity. The fine structuring attained enables dramatically accelerated diffusion and enhances the electrochemical sensing performance for disease-specific biomolecules. As a proof-of-concept, the as-fabricated glass-grown mAu film demonstrates high sensitivity in electrochemical detection of SARS-CoV-2-specific RNA with a limit of detection (LoD) as low as 1 attomolar (aM).

In the present work, a novel electrochemical DNA sensor was designed to detect L. monocytogenes. Two different gene fragments were selected for the sensor design. One is a 702 bp long fragment of the hlyA gene, encoding the synthesis of listeriolysin O toxin, which is unique only to pathogenic strains of L. monocytogenes and is essential for pathogenicity. The other is a 209 bp long fragment of the 16 S RNA gene found in all species of the Listeria genus. As the working electrode, the pencil graphite electrode was modified in various ways (activated or covered with polypyrrole), and six different combinations were constituted using three types of the modified working electrode and two different gene fragments. The developed system is based on differential pulse voltammetric transduction of guanine oxidation after hybridization between the selected gene fragment (38 µg/mL) and the selected fragment-specific inosine-modified probe (1.8 µmol/L) immobilized on a pencil graphite electrode surface. The comparison of all combinations demonstrates that the best results are obtained with the combination formed from a polypyrrole-coated pencil graphite electrode (prepared at 2 scans) and 702 bp fragment of the hlyA gene. The analysis time is less than 1 hour, and the necessary DNA concentrations for the analysis have been determined as 8.2 × 10-11 M DNA and 2.7 × 10-10 M DNA respectively, for the hlyA gene and 16 S RNA gene.

Point-of-care (POC) devices have become rising stars in the biosensing field, aiming at prognosis and diagnosis of diseases with a positive impact on the patient but also on healthcare and social care systems. Putting the patient at the center of interest requires the implementation of noninvasive technologies for collecting biofluids and the development of wearable platforms with integrated artificial intelligence-based tools for improved analytical accuracy and wireless readout technologies. Many electrical and electrochemical transducer technologies have been proposed for POC-based sensing, but several necessitate further development before being widely deployable. This review focuses on recent innovations in electrochemical and electrical biosensors and their growth opportunities for nanotechnology-driven multidisciplinary approaches. With a focus on analytical aspects to pave the way for future electrical/electrochemical diagnostics tests, current limitations and drawbacks as well as directions for future developments are highlighted.

The number of innovative applications for DNA nowadays is growing quickly. Its use as a nanowire or electrochemical biosensor leads to the need for a deep understanding of the charge-transfer process along the strand, as well as its redox properties. These features are computationally simulated and analyzed in detail throughout this work by combining molecular dynamics, multilayer schemes, and the Marcus theory. One-electron oxidation potential and hole delocalization have been analyzed for six DNA double strands that cover all possible binary combinations of nucleotides. The results have revealed that the one-electron oxidation potential decreases with respect to the single-stranded DNA, giving evidence that the greater rigidity of a double helix induces an increase in the capacity of storing the positive charge generated upon oxidation. In addition, the hole is mainly stored in nucleobases with large reducer character, i.e., purines, especially when those are arranged in a stacked configuration in the same strand. From the computational point of view, the sampling needed to describe biological systems implies a significant computational cost. Here, we show that a small number of representative conformations generated by clustering analysis provides accurate results when compared with those obtained from sampling, reducing considerably the computational cost.

Simultaneous detection of multiple tumor biomarkers with a simple and low-cost assay is crucial for early cancer detection and diagnosis. Herein, we presented a low-cost and simple assay for multiplexed detection of tumor biomarkers using a spatially separated electrodes strategy. The sensor is fabricated based on a metal-free thiol-yne click reaction, which is mediated by visible light, on commercially available indium tin oxide (ITO) electrodes. Four biomarkers, including p53 DNA, Brca2 DNA, K-ras DNA, and MicroRNA-204 RNA, were used as model analytes, and the corresponding oligonucleotide probes were modified on the desired electrode units sequentially with 530 nm irradiation light in the presence of photosensitizer Eosin Y. By this visible light-mediated coupling reaction, oligonucleotide probe densities of up to 9.2 ± 0.7 × 1010 molecules/cm2 were readily obtained on the ITO electrode surface. The proposed multiplexed E-NA sensor could detect four different nucleic acid targets concurrently without crosstalk among adjacent electrodes and was also successfully applied for detecting targets in a 20% fetal calf serum sample. The detection limits for p53 DNA, Brca2 DNA, K-ras DNA, and MicroRNA-204 RNA were 0.72 nM, 0.97 nM, 2.15 nM, and 1.73 nM, respectively. The developed approach not only has a great potential for developing cost-effective biosensors on affordable substrates for nucleic acid target detection, but also be easily extended to detect other targets by modifying the specific oligonucleotide probes anchored on the electrode.

Due to their life cycle, viruses can disrupt the metabolism of their hosts, causing diseases. If we want to disrupt their life cycle, it is necessary to identify their presence. For this purpose, it is possible to use several molecular-biological and bioanalytical methods. The reference selection was performed based on electronic databases (2020-2023). This review focused on electrochemical methods with high sensitivity and selectivity (53% voltammetry/amperometry, 33% impedance, and 12% other methods) which showed their great potential for detecting various viruses. Moreover, the aforementioned electrochemical methods have considerable potential to be applicable for care-point use as they are portable due to their miniaturizability and fast speed analysis (minutes to hours), and are relatively easy to interpret. A total of 2011 articles were found, of which 86 original papers were subsequently evaluated (the majority of which are focused on human pathogens, whereas articles dealing with plant pathogens are in the minority). Thirty-two species of viruses were included in the evaluation. It was found that most of the examined research studies (77%) used nanotechnological modifications. Other ones performed immunological (52%) or genetic analyses (43%) for virus detection. 5% of the reports used peptides to increase the method's sensitivity. When evaluable, 65% of the research studies had LOD values in the order of ng or nM. The vast majority (79%) of the studies represent proof of concept and possibilities with low application potential and a high need of further research experimental work.

Glucose is an important biomarker for diagnosing and prognosing various diseases, including diabetes and hypoglycemia, which can have severe side effects, symptoms, and even lead to death in patients. As a result, there is a need for quick and economical glucose level measurements to help identify those at potential risk. With the increase in smartphone users, portable smartphone glucose sensors are becoming popular. In this paper, we present a disposable microfluidic glucose sensor that accurately and rapidly quantifies glucose levels in human urine using a combination of colorimetric analysis and computer vision. This glucose sensor implements a disposable microfluidic device based on medical-grade tapes and glucose analysis strips on a glass slide integrated with a custom-made polydimethylsiloxane (PDMS) micropump that accelerates capillary flow, making it economical, convenient, rapid, and equipment-free. After absorbing the target solution, the disposable device is slid into the 3D-printed main chassis and illuminated exclusively with Light Emitting Diode (LED) illumination, which is pivotal to color-sensitive experiments. After collecting images, the images are imported into the algorithm to measure the glucose levels using computer vision and average RGB values measurements. This article illustrates the impressive accuracy and consistency of the glucose sensor in quantifying glucose in sucrose water. This is evidenced by the close agreement between the computer vision method used by the sensor and the traditional method of measuring in the biology field, as well as the small variation observed between different sensor performances. The exponential regression curve used in the study further confirms the strong relationship between glucose concentrations and average RGB values, with an R-square value of 0.997 indicating a high degree of correlation between these variables. The article also emphasizes the potential transferability of the solution described to other types of assays and smartphone-based sensors.

Organic metallic nanohybrids (NHs), in which many small metal nanoparticles are encapsulated within a conductive polymer matrix, are useful as sensitive electrochemical labels because the constituents produce characteristic oxidation current responses. Gold NHs, consisting of gold nanoparticles and poly(m-toluidine), and copper NHs, consisting of copper nanoparticles and polyaniline, did not interfere with each other in terms of the electrochemical signals obtained on the same electrode. Antibodies were introduced into these NHs to function as electrochemical labels for targeting specific bacteria. Electrochemical measurements using screen-printed electrodes dry-fixed with NH-labeled bacterial cells enabled the estimation of bacterial species and number within minutes, based on the distinct current response of the labels. Our proposed method achieved simultaneous detection of enterohemorrhagic Escherichia coli and Staphylococcus aureus in a real sample. These NHs will be powerful tools as electrochemical labels and are expected to be useful for rapid testing in food and drug-related manufacturing sites.

Recent decades have experienced tough situations due to the lack of reliable diagnostic facilities. The most recent cases occurred during the pandemic, where researchers observed the lack of diagnostic facilities with precision. Microorganisms and viral disease's ability to escape diagnosis has been a global challenge. DNA always has been a unique moiety with a strong and precise base-paired structure. DNA in human and foreign particles makes identification possible through base pairing. Since then, researchers have focused heavily on designing diagnostic assays targeting DNA in particular. Moreover, DNA nanotechnology has contributed vastly to designing composite nanomaterials by combining DNA/nucleic acids with functional nanomaterials and inorganic nanoparticles exploiting their physicochemical properties. These nanomaterials often exhibit unique or enhanced properties due to the synergistic activity of the many components. The capabilities of DNA and additional nanomaterials have shown the combination of robust and advanced tailoring of biosensors. Preceding findings state that the conventional strategies have exhibited certain limitations such as a low range of target detection, less biodegradability, subordinate half-life, and high susceptibility to microenvironments; however, a DNA-nanomaterial-based biosensor has overcome these limitations meaningfully. Additionally, the unique properties of nucleic acids have been studied extensively due to their high signal conduction abilities. Here, we review recent studies on DNA-nanomaterial-based biosensors, their mechanism of action, and improved/updated strategies in vivo and in situ. Furthermore, this review highlights the recent methodologies on DNA utilization to exploit the interfacial properties of nanomaterials in DNA sensing. Lastly, the review concludes with the limitations/challenges and future directions.

The abnormal expression of human DNA methyltransferases (DNMTs) is closely related with the occurrence and development of a wide range of human cancers. DNA (cytosine-5)-methyltransferase-1 (DNMT1) is the most abundant human DNA methyltransferase and is mainly responsible for genomic DNA methylation patterns. Abnormal expression of DNMT1 has been found in many kinds of tumors, and DNMT1 has become a valuable target for the diagnosis and drug therapy of diseases. Nowadays, DNMT1 has been found to be involved in multiple cancers such as pancreatic cancer, breast cancer, bladder cancer, lung cancer, gastric cancer and other cancers. In order to achieve early diagnosis and for scientific research, various analytical methods have been developed for qualitative or quantitative detection of low-abundance DNMT1 in biological samples and human tumor cells. Herein, we provide a brief explication of the research progress of DNMT1 involved in various cancer types. In addition, this review focuses on the types, principles, and applications of DNMT1 detection methods, and discusses the challenges and potential future directions of DNMT1 detection.

DNA nanotechnology deals with the design of non-naturally occurring DNA nanostructures that can be used in biotechnology, medicine, and diagnostics. In this study, we introduced a nucleic acid five-way junction (5WJ) structure for direct electrochemical analysis of full-length biological RNAs. To the best of our knowledge, this is the first report on the interrogation of such long nucleic acid sequences by hybridization probes attached to a solid support. A hairpin-shaped electrode-bound oligonucleotide hybridizes with three adaptor strands, one of which is labeled with methylene blue (MB). The four strands are combined into a 5WJ structure only in the presence of specific DNA or RNA analytes. Upon interrogation of a full-size 16S rRNA in the total RNA sample, the electrode-bound MB-labeled 5WJ association produces a higher signal-to-noise ratio than electrochemical nucleic acid biosensors of alternative design. This advantage was attributed to the favorable geometry on the 5WJ nanostructure formed on the electrode's surface. The 5WJ biosensor is a cost-efficient alternative to the traditional electrochemical biosensors for the analysis of nucleic acids due to the universal nature of both the electrode-bound and MB-labeled DNA components.

The fast and precise selective sensing of DNA nucleobases is a long-pursued method that can lead to huge advances in the field of genomics and have an impact on aspects such as the prevention of diseases, health enhancement, and, in general, all types of medical treatments. We present here a new type of nanoscale sensor based on carbon nanotubes with a specific geometry that can discriminate the type of nucleobase and also its angle of orientation. The proper differentiation of nucleobases is essential to clearly sequence DNA chains, while angular discrimination is key to improving the sensing selectivity. We perform first-principle and quantum transport simulations to calculate the transmission, conductance, and current of the nanotube-based nanoscale sensor in the presence of the four nucleotides (A, C, G, and T), each of them rotated 0, 90, 180, or 270°. Our results show that this system is able to effectively discriminate between the four nucleotides and their angle of orientation. We explain these findings in terms of the interaction between the phosphate group of the nucleotide and the nanotube wall. The phosphate specifically distorts the electronic structure of the nanotube depending on the distance and the orientation and leads to nontrivial changes in the transmission. This work provides a method for finer and more precise sequential DNA chains.

Fibrinogen as a major inflammation marker and blood coagulation factor has a direct impact on the health of humanity. The variations in fibrinogen content lead to risky conditions such as bleeding and cardiovascular diseases. So, accurate methods for monitoring of this glycoprotein are of high importance. The conventional methods, such as the Clauss method, are time consuming and require highly specialized expert analysts. The development of fast, simple, easy to use, and inexpensive methods is highly desired. In this way, biosensors have gained outstanding attention since they offer means for performing analyses at the points-of-care using self-testing devices, which can be applied outside of clinical laboratories or hospital. This review indicates that different electrochemical and optical sensors have been successfully implemented for the detection of fibrinogen under normal levels of fibrinogen in plasma. The biosensors for the detection of fibrinogen have been designed based on the quartz crystal microbalance, field-effect transistor, electrochemical impedance spectroscopy, amperometry, surface plasmon resonance, localized surface plasmon resonance, and colorimetric techniques. Also, this review demonstrates the utility of the application of nanoparticles in different detection techniques.

Mechanical forces play an important role in cellular communication and signaling. We developed in this study novel electrochemical DNA-based force sensors for measuring cell-generated adhesion forces. Two types of DNA probes, i.e., tension gauge tether and DNA hairpin, were constructed on the surface of a smartphone-based electrochemical device to detect piconewton-scale cellular forces at tunable levels. Upon experiencing cellular tension, the unfolding of DNA probes induces the separation of redox reporters from the surface of the electrode, which results in detectable electrochemical signals. Using integrin-mediated cell adhesion as an example, our results indicated that these electrochemical sensors can be used for highly sensitive, robust, simple, and portable measurement of cell-generated forces.

DNA helicase activity is essential for the vital DNA metabolic processes of recombination, replication, transcription, translation, and repair. Recently, an unexpected, rapid exponential ATP-stimulated DNA unwinding rate was observed from an Archaeoglobus fulgidus helicase (AfXPB) as compared to the slower conventional helicases from Sulfolobus tokodaii, StXPB1 and StXPB2. This unusual rapid activity suggests a "molecular wrench" mechanism arising from the torque applied by AfXPB on the duplex structure in transitioning from open to closed conformations. However, much remains to be understood. Here, we investigate the concentration dependence of DNA helicase binding and ATP-stimulated kinetics of StXPB2 and AfXPB, as well as their binding and activity in Bax1 complexes, via an electrochemical assay with redox-active DNA monolayers. StXPB2 ATP-stimulated activity is concentration-independent from 8 to 200 nM. Unexpectedly, AfXPB activity is concentration-dependent in this range, with exponential rate constants varying from seconds at concentrations greater than 20 nM to thousands of seconds at lower concentrations. At 20 nM, rapid exponential signal decay ensues, linearly reverses, and resumes with a slower exponential decay. This change in AfXPB activity as a function of its concentration is rationalized as the crossover between the fast molecular wrench and slower conventional helicase modes. AfXPB-Bax1 inhibits rapid activity, whereas the StXPB2-Bax1 complex induces rapid kinetics at higher concentrations. This activity is rationalized with the crystal structures of these complexes. These findings illuminate the different physical models governing molecular wrench activity for improved biological insight into a key factor in DNA repair.

The study of DNA processes is essential to understand not only its intrinsic biological functions but also its role in many innovative applications. The use of DNA as a nanowire or electrochemical biosensor leads to the need for a deep investigation of the charge transfer process along the strand as well as of the redox properties. In this contribution, the one-electron oxidation potential and the charge delocalization of the hole formed after oxidation are computationally investigated for different heterogeneous single-stranded DNA strands. We have established a two-step protocol: (i) molecular dynamics simulations in the frame of quantum mechanics/molecular mechanics (QM/MM) were performed to sample the conformational space; (ii) energetic properties were then obtained within a QM1/QM2/continuum approach in combination with the Marcus theory over an ensemble of selected geometries. The results reveal that the one-electron oxidation potential in the heterogeneous strands can be seen as a linear combination of that property within the homogeneous strands. In addition, the hole delocalization between different nucleobases is, in general, small, supporting the conclusion of a hopping mechanism for charge transport along the strands. However, charge delocalization becomes more important, and so does the tunneling mechanism contribution, when the reducing power of the nucleobases forming the strand is similar. Moreover, charge delocalization is slightly enhanced when there is a correlation between pairs of some of the interbase coordinates of the strand: twist/shift, twist/slide, shift/slide, and rise/tilt. However, the internal structure of the strand is not the predominant factor for hole delocalization but the specific sequence of nucleotides that compose the strand.

Glucose serves as a pivotal biomarker crucial for the monitoring and diagnosis of a spectrum of medical conditions, encompassing hypoglycemia, hyperglycemia, and diabetes, all of which may precipitate severe clinical manifestations in individuals. As a result, there is a growing demand within the medical domain for the development of rapid, cost-effective, and user-friendly diagnostic tools. In this research article, we introduce an innovative glucose sensor that relies on microfluidic devices meticulously crafted from disposable, medical-grade tapes. These devices incorporate glucose urine analysis strips securely affixed to microscope glass slides. The microfluidic channels are intricately created through laser cutting, representing a departure from traditional cleanroom techniques. This approach streamlines production processes, enhances cost-efficiency, and obviates the need for specialized equipment. Subsequent to the absorption of the target solution, the disposable device is enclosed within a 3D-printed housing. Image capture is seamlessly facilitated through the use of a smartphone camera for subsequent colorimetric analysis. Our study adeptly demonstrates the glucose sensor's capability to accurately quantify glucose concentrations within sucrose solutions. This is achieved by employing an exponential regression model, elucidating the intricate relationship between glucose concentrations and average RGB (Red-Green-Blue) values. Furthermore, our comprehensive analysis reveals minimal variation in sensor performance across different instances. Significantly, this study underscores the potential adaptability and versatility of our solution for a wide array of assay types and smartphone-based sensor systems, making it particularly promising for deployment in resource-constrained settings and undeveloped countries. The robust correlation established between glucose concentrations and average RGB values, substantiated by an impressive R-square value of 0.98709, underscores the effectiveness and reliability of our pioneering approach within the medical field.

The combination of gene therapy and immunotherapy concepts, along recent advances in DNA nanotechnology, have the potential to provide important tools for cancer therapies. We present the development of stimuli-responsive microcapsules, loaded with a viral immunogenetic agent, harnessing the immune response against the Coronavirus Disease 2019, COVID-19, to selectively attack liver cancer cells (hepatoma) or recognize breast cancer or hepatoma, by expression of green fluorescence protein, GFP. The pH-responsive microcapsules, modified with DNA-tetrahedra nanostructures, increased hepatoma permeation by 50 %. Incorporation of a GFP-encoding lentivirus vector inside the tumor-targeting pH-stimulated miRNA-triggered and Alpha-fetoprotein-dictated microcapsules enables the demonstration of neoplasm selectivity, with approximately 5,000-, 8,000- and 50,000-fold more expression in the cancerous cells, respectively. The incorporation of the SARS-CoV-2 spike protein in the gene vector promotes specific recognition of the immune-evading hepatoma by the COVID-19-analogous immune response, which leads to cytotoxic and inflammatory activity, mediated by serum components taken from vaccinated or recovered COVID-19 patients, resulting in effective elimination of the hepatoma (>85 % yield).

While natural oligonucleotides (ONs) are increasingly used as therapeutic and diagnostic tools, they still face certain challenges such as low resistance to enzymatic degradation, potential immunogenicity, and delivery issues, which can limit their applications. Peptide Nucleic Acids (PNAs) are promising alternatives due to their high affinity for DNA and RNA, the high resistance to enzymatic degradation, and the easy introduction of a wide range of potential modifications. Chemical modifications that enable the covalent targeting of specific DNA and RNA strands offer additional advantages, including enhanced potency. The current study focuses on the utilization of furan-PNAs as pro-reactive probe systems and their applications to DNA and RNA targeting. Specifically, in this methodological paper, we provide practical insights into the design, synthesis, and application of furan-containing PNA probes for achieving efficient PNA-DNA and PNA-RNA interstrand crosslinking (ICL), as well as ON-templated PNA-PNA ligation systems. Furthermore, we discuss the applications of these probes in targeting DNA secondary structures, such as G-quadruplexes and i-motifs, target pull-down assays, and on-surface detection.

A modular, multi-purpose, and cost-effective electrochemical biosensor based on a five-stranded four-way junction (5S-4WJ) system was developed for SARS-CoV-2 (genes S and N) and Influenza A virus (gene M) detection. The 5S-4WJ structure consists of an electrode-immobilized universal stem-loop (USL) strand, two auxiliary DNA strands, and a universal methylene blue redox strand (UMeB). This design allows for the detection of specific nucleic acid sequences using square wave voltammetry (SWV). The sequence-specific auxiliary DNA strands (m and f) ensure selectivity of the biosensor for target recognition utilizing the same USL and UMeB components. An important feature of this biosensor is the ability to reuse the USL-modified electrodes to detect the same or alternative targets in new samples. This is accomplished by a simple procedure involving rinsing the electrodes with water to disrupt the 5S-4WJ structure and subsequent re-hybridization of the USL strand with the appropriate set of strands for a new analysis. The biosensor exhibited minimal loss in signal after rehybridization, demonstrating its potential as a viable multiplex assay for both current and future pathogens, with a low limit of quantification (LOQ) of as low as 17 pM.

Oligonucleotide-functionalized graphene biosensors show immense promise for use as label-free point of care devices for detection of nucleic acid biomarkers at clinically relevant levels. Graphene-based nucleic acid sensors can be fabricated at low cost and have been shown to reach limits of detection in the attomolar range. Here we demonstrate devices functionalized with 22mer or 8omer DNA probes are capable of detecting full length genomic HIV-1 subtype B RNA, with a limit of detection below 1 aM in nuclease free water. We also show that these sensors are suitable for detection directly in Qiazol lysis reagent, again with a limit of detection below 1 aM for both 22mer and 8omer probes.

Oligonucleotide-based therapeutics have attracted attention as an emerging modality that includes the modulation of genes and their binding proteins related to diseases, allowing us to take action on previously undruggable targets. Since the late 2010s, the number of oligonucleotide medicines approved for clinical uses has dramatically increased. Various chemistry-based technologies have been developed to improve the therapeutic properties of oligonucleotides, such as chemical modification, conjugation, and nanoparticle formation, which can increase nuclease resistance, enhance affinity and selectivity to target sites, suppress off-target effects, and improve pharmacokinetic properties. Similar strategies employing modified nucleobases and lipid nanoparticles have been used for developing coronavirus disease 2019 mRNA vaccines. In this review, we provide an overview of the development of chemistry-based technologies aimed at using nucleic acids for developing therapeutics over the past several decades, with a specific emphasis on the structural design and functionality of chemical modification strategies.

Electrochemical DNA sensors can be operated in either static or flow-based detection schemes. In static schemes, manual washing steps are still necessary, resulting in a tedious and time-consuming process. In contrast, in flow-based electrochemical sensors, the current response is collected when the solution flows through the electrode continuously. However, the drawback of such a flow system is the low sensitivity due to the limited time for the interaction between the capturing element and the target. Herein, we propose a novel electrochemical capillary-driven microfluidic DNA sensor to combine the advantages of static and flow-based electrochemical detection systems into a single device by incorporating burst valve technology. The microfluidic device with a two-electrode configuration was applied for the simultaneous detection of two different DNA markers, human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV) cDNA, via the specific interaction between pyrrolidinyl peptide nucleic acids (PNA) probes and the DNA target. The integrated system, while requiring a small sample volume (7 μL for each sample loading port) and less analysis time, achieved good performance in terms of the limits of detection (LOD) (3SD_blank/slope) and quantification (LOQ) (10SD_blank/slope) at 1.45 nM and 4.79 nM for HIV and 1.20 nM and 3.96 nM for HCV, respectively. The simultaneous detection of HIV-1 and HCV cDNA prepared from human blood samples showed results that are in complete agreement with the RT‒PCR assay. The results qualify this platform as a promising alternative for the analysis of either HIV-1/HCV or coinfection that can be easily adapted for other clinically important nucleic acid-based markers.

Throughout the world, oral cancer is a common and aggressive malignancy with a high risk of morbidity, mortality, and recurrence. The importance of early detection in cancer prevention and disease treatment cannot be overstated. Conventional therapeutic strategies have minor difficulties but considerable side effects and unfavourable consequences in clinical applications. Hence, there is a requirement for effective ways for early detection and treatment of oral cancer. At present, numerous forms of nanoparticles have piqued researchers' interest as a potentially useful tool for diagnostic probes and medicinal devices. Because of their inherent physicochemical properties and customizable surface modification, they are able to circumvent some of restrictions and accomplish the intended diagnostic and therapeutic impact. Nanotechnology is a unique field that has revolutionised the industry and is paving the way for new treatments for oral cancer. It can help with a better diagnosis with less harmful substances and is setting current guidelines for treatment. The use of nanotechnology in cancer diagnosis, therapy, and care improves clinical practise dramatically. The different types of nanoparticles that have been developed for the diagnosis and therapy of oral cancers will be covered in this study. The difficulties and potential uses of nanoparticles in the treatment and diagnosis of oral cancer are then highlighted. In order to emphasise existing difficulties and potential remedies for oral cancer, a prospective view of the future is also provided.

DNA has been demonstrated as a powerful platform for the construction of inorganic nanoparticles (NPs) into complex three-dimensional assemblies. Despite extensive research, the physical fundamental details of DNA nanostructures and their assemblies with NPs remain obscure. Here, we report the identification and quantification of the assembly details of programmable DNA nanotubes with monodisperse circumferences of a 4, 5, 6, 7, 8, or 10 DNA helix and their pearl-necklace-like assemblies with ultrasmall gold nanoparticles, Au₂₅ nanoclusters (AuNCs), liganded by -S(CH₂)_nNH₃⁺ (n = 3, 6, 11). The flexibilities of DNA nanotubes, analyzed via statistical polymer physics analysis through atomic force microscopy (AFM), demonstrate that ∼2.8 power exponentially increased with the DNA helix number. Moreover, the short-length liganded AuS(CH₂)₃NH₃⁺ NCs were observed to be able to form pearl-necklace-like DNA-AuNC assemblies stiffened than neat DNA nanotubes, while long-length liganded AuS(CH₂)₆NH₃⁺ and AuS(CH₂)₁₁NH₃⁺ NCs could fragment DNA nanotubular structures, indicating that DNA-AuNC assembling can be precisely manipulated by customizing the hydrophobic domains of the AuNC nanointerfaces. We prove the advantages of polymer science concepts in unraveling useful intrinsic information on physical fundamental details of DNA-AuNC assembling, which facilitates DNA-metal nanocomposite construction.

The effective charge of highly charged polyelectrolytes is significantly lowered by a condensation of counterions. This effect is more pronounced for divalent ions. Here we present a study of the counterion condensation to dendritic polyglycerol sulfate (dPGS) that consists of a hydrophilic dendritic scaffold onto which sulfate groups are appended. The interactions between the dPGS and divalent ions (Mg²⁺ and Ca²⁺) were analyzed using isothermal titration calorimetry (ITC) and showed no ion specificity upon binding, but clear competition between the monovalent and divalent ions. Our findings, in line with the latest theoretical studies, demonstrate that a large fraction of the monovalent ions is sequentially replaced with the divalent ions.