Emerging role of exosomal microRNA in liver cancer in the era of precision medicine; potential and challenges.

Exosomal microRNAs (miRNAs) have great potential in the fight against hepatocellular carcinoma (HCC), the fourth most common cause of cancer-related death worldwide. In this study, we explored the various applications of these small molecules while analyzing their complex roles in tumor development, metastasis, and changes in the tumor microenvironment. We also discussed the complex interactions that exist between exosomal miRNAs and other non-coding RNAs such as circular RNAs, and show how these interactions coordinate important biochemical pathways that propel the development of HCC. The possibility of targeting exosomal miRNAs for therapeutic intervention is paramount, even beyond their mechanistic significance. We also highlighted their growing potential as cutting-edge biomarkers that could lead to tailored treatment plans by enabling early identification, precise prognosis, and real-time treatment response monitoring. This thorough analysis revealed an intricate network of exosomal miRNAs lead to HCC progression. Finally, strategies for purification and isolation of exosomes and advanced biosensing techniques for detection of exosomal miRNAs are also discussed. Overall, this comprehensive review sheds light on the complex web of exosomal miRNAs in HCC, offering valuable insights for future advancements in diagnosis, prognosis, and ultimately, improved outcomes for patients battling this deadly disease.

Biosensing of Alpha-Fetoprotein: A Key Direction toward the Early Detection and Management of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is currently one of the most prevalent cancers worldwide. Associated risk factors include, but are not limited to, cirrhosis and underlying liver diseases, including chronic hepatitis B or C infections, excessive alcohol consumption, nonalcoholic fatty liver disease (NAFLD), and exposure to chemical carcinogens. It is crucial to detect this disease early on before it metastasizes to adjoining parts of the body, worsening the prognosis. Serum biomarkers have proven to be a more accurate diagnostic tool compared to imaging. Among various markers such as nucleic acids, circulating genetic material, proteins, enzymes, and other metabolites, alpha-fetoprotein (AFP) is a protein marker primarily used to diagnose HCC. However, current methods need a large sample and carry a high cost, among other challenges, which can be improved using biosensing technology. Early and accurate detection of AFP can prevent severe progression of the disease and ensure better management of HCC patients. This review sheds light on HCC development in the human body. Afterward, we outline various types of biosensors (optical, electrochemical, and mass-based), as well as the most relevant studies of biosensing modalities for non-invasive monitoring of AFP. The review also explains these sensing platforms, detection substrates, surface modification agents, and fluorescent probes used to develop such biosensors. Finally, the challenges and future trends in routine clinical analysis are discussed to motivate further developments.

Thread-powered cell lysis and isotachophoresis: unlocking microbial DNA for diverse molecular applications.

Central to the domain of molecular biology resides the foundational process of DNA extraction and purification, a cornerstone underpinning a myriad of pivotal applications. In this research, we introduce a DNA extraction and purification technique leveraging polypropylene (PP) threads. The process commences with robust cell lysis achieved through the vigorous agitation of interwoven PP threads. The friction between the threads facilitates cell lysis especially those microbes having tough cell wall. For purification of DNA, thread-based isotachophoresis was employed which makes the whole process swift and cost-effective. Lysed cell-laden threads were submerged in a trailing electrolyte which separated DNA from other cellular contents. The process was performed with a tailored ITP device. An electric field directs DNA, cell debris, trailing electrolyte, and leading electrolyte toward the anode. Distinct ion migration resulted in DNA concentrating on the PP thread's anode-proximal region. The SYBR green dye is used to visualize DNA as a prominent green zone under blue light. The purified DNA exhibits high purity levels of 1.82 ± 0.1 (A260/A280), making it suitable for various applications aiming at nucleic acid detection.

Hair and Nail-On-Chip for Bioinspired Microfluidic Device Fabrication and Biomarker Detection.

The advent of biosensors has tremendously increased our potential of identifying and solving important problems in various domains, ranging from food safety and environmental analysis, to healthcare and medicine. However, one of the most prominent drawbacks of these technologies, especially in the biomedical field, is to employ conventional samples, such as blood, urine, tissue extracts and other body fluids for analysis, which suffer from the drawbacks of invasiveness, discomfort, and high costs encountered in transportation and storage, thereby hindering these products to be applied for point-of-care testing that has garnered substantial attention in recent years. Therefore, through this review, we emphasize for the first time, the applications of switching over to noninvasive sampling techniques involving hair and nails that not only circumvent most of the aforementioned limitations, but also serve as interesting alternatives in understanding the human physiology involving minimal costs, equipment and human interference when combined with rapidly advancing technologies, such as microfluidics and organ-on-a-chip to achieve miniaturization on an unprecedented scale. The coalescence between these two fields has not only led to the fabrication of novel microdevices involving hair and nails, but also function as robust biosensors for the detection of biomarkers, chemicals, metabolites and nucleic acids through noninvasive sampling. Finally, we have also elucidated a plethora of futuristic innovations that could be incorporated in such devices, such as expanding their applications in nail and hair-based drug delivery, their potential in serving as next-generation wearable sensors and integrating these devices with machine-learning for enhanced automation and decentralization.

Tuning Hydrophobicity of Paper Substrates for Effective Colorimetric detection of Glucose and Nucleic acids.

This study investigated the colorimetric response of standard glucose, serum glucose, and nucleic acid assays on various paper surfaces with different wettability, including hydrophilic, hydrophobic, and nearly superhydrophobic surfaces. Water contact angles (WCA) formed by water droplets on each surface were measured using ImageJ software. The hydrophilic surface showed no contact angle, while the hydrophobic and nearly superhydrophobic surfaces exhibited contact angles of 115.667° and 133.933°, respectively. The colorimetric sensitivity of the standard glucose assay was analyzed on these surfaces, revealing enhanced sensitivity on the nearly superhydrophobic surface due to the high molecular crowding effect owing to its non-wetting behavior and eventually confined reaction product at the sample loading zone. The hydrophobic nature of the surface restricts the spreading and diffusion of the reaction product, leading to a controlled and localized concentration of the assay product leading to moderate colorimetric intensity. On the other hand, the hydrophilic surface showed the least enhancement in colorimetric sensitivity; this is attributed to the high wettability of the hydrophilic surface causing the reaction product to spread extensively, resulting in a larger area of dispersion and consequently a lower colorimetric intensity. The measured limit of detection (LOD) for nucleic acid on nearly superhydrophobic surfaces was found to be 16.15 ng/µL, which was almost four-fold lower than on hydrophilic surfaces (60.08 ng/µL). Additionally, the LODs of standard glucose and clinical serum samples were two-fold lower on nearly superhydrophobic surfaces compared to hydrophilic surfaces. Our findings clearly highlight the promising potential of utilizing superhydrophobic surfaces to significantly enhance colorimetric sensitivity in paper-based diagnostic applications. This innovative approach holds promise for advancing point-of-care diagnostics and improving disease detection in resource-limited settings.

DNA compaction enhances the sensitivity of fluorescence-based nucleic acid assays: a game changer in point of care sensors?

Fluorescence-based nucleic acid assays frequently exhibit a feeble signal at low analyte concentrations, necessitating complex, expensive methods such as the development of sequence-specific oligo tags, molecular beacons, and chemical modifications to maintain high detection sensitivity. Hence, there is growing interest in accomplishing fluorescence enhancement in nucleic acid assays using robust and cost-effective strategies. The study exploits the use of two compaction agents, PEG 8000 and CTAB, to compact the ITS-2 amplicon of the fungus Candida albicans and evaluates the effect of both of these agents on the fluorescence intensity of SYTO-9 labelled nucleic acids. Conventional fluorometric measurements showed that both CTAB and PEG 8000 enhanced the emission intensity by ∼1.2- and 2-fold, respectively. Furthermore, we leveraged paper-based spot tests and distance-based assays to validate the effect of DNA compaction for enhancing sensitivity in the point-of-care context. The spot assay performed on paper with compacted samples showed an increase in the emission intensity of SYTO-9 and this was manifested by an elevated G channel intensity in the order of PEG 8000 compacted > CTAB compacted > amplified. Moreover, in the distance-based assay, the PEG 8000 compacted sample was found to migrate farther compared to CTAB compacted and amplified DNA samples at amplicon concentrations, 15 µg ml-1 and 39.65 µg ml-1. The limit of detection (LOD) for PEG 8000 and CTAB compacted samples on both paper-spot and distance-based assays were found to be 0.4 µg ml-1 and 0.5 µg ml-1, respectively. Hence our work provides an overview of employing DNA compaction as an approach for enhancing the sensitivity of fluorescence-based point-of-care nucleic acid assays without the need for cumbersome sensitivity enhancement methods.

Paper-based dots and smartphone for detecting counterfeit country eggs.

Herein, we present for the first time, the employment of paper-based devices for rapidly differentiating original country eggs from the plain broiler eggs that have been coated with tea to appear as the former. The devices leverage two types of phenomena involving the phenols present in tea in precisely 5 min, namely precipitation, which produces a well-defined dark bluish precipitate on the surface of the counterfeit country eggs or tea-coated broiler eggs and de-coloration, wherein the dried layer of tea coating present on the surface of the dummy country eggs get dissolved, thereby revealing the white colour of the plain broiler egg shell. To reduce the subjectivity, a smartphone application 'Eggo' has been developed which is capable of detecting the spots produced by both the methods using mobile's camera. Fourier Transform Infrared Spectroscopy (FTIR) analysis was performed to study the changes occurring on the shell surface. Such sophisticated yet simple technologies will revolutionize food fraud analysis.

A Hypothetical Approach to Concentrate Microorganisms from Human Urine Samples Using Paper-Based Adsorbents for Point-of-Care Molecular Assays.

This hypothesis demonstrates that the efficiency of loop-mediated isothermal amplification (LAMP) for nucleic acid detection can be positively influenced by the preconcentration of microbial cells onto hydrophobic paper surfaces. The mechanism of this model is based on the high affinity of microbes towards hydrophobic surfaces. Extensive studies have demonstrated that hydrophobic surfaces exhibit enhanced bacterial and fungal adhesion. By exploiting this inherent affinity of hydrophobic paper substrates, the preconcentration approach enables the adherence of a greater number of target cells, resulting in a higher concentration of target templates for amplification directly from urine samples. In contrast to conventional methods, which often involve complex procedures, this approach offers a simpler, cost-effective, and user-friendly alternative. Moreover, the integration of cell adhesion, LAMP amplification, and signal readout within paper origami-based devices can provide a portable, robust, and highly efficient platform for rapid nucleic acid detection. This innovative hypothesis holds significant potential for point-of-care (POC) diagnostics and field surveillance applications. Further research and development in this field will advance the implementation of this technology, contributing to improved healthcare systems and public health outcomes.

CRISPR/Cas technology: Opportunities for phytopathogenic viruses detection.

Detection and monitoring of viruses are essential for healthy plants and prosperity. Recent development in CRISPR/Cas system in diagnosis has open an avenue well suited for pathogen detection. Variety of CRISPR associated proteins are being discovered, suggesting array of application and detection strategies in diagnosis. Phytopathogenic viruses are diverse with respect to their nucleic acid compositions, which presents a challenge in developing a single device applicable for almost all viruses. The review describes about the efficient use of CRISPR/Cas Technology in diagnosis, such as SHERLOCK, DETECTR and SATORI. These methods are different in their characteristic to identify specific nucleic acids and processing the detectable signals. These technologies are in their infancy and lot of scope is there to develop commercial kits. Plant tissue culture-based industries, climate control green houses, indoor cultivation facilities etc. has been considered as few examples. This review will be beneficial for researchers seeking to develop detection mechanism based on CRISPR/Cas technology. The outcome in the form of cost-effective detection of viruses will be boon for agro-based industries, which are facing challenges through virus contamination.

Embellishing 2-D MoS2 Nanosheets on Lotus Thread Devices for Enhanced Hydrophobicity and Antimicrobial Activity.

Herein, we report cellulose-based threads from Indian sacred Lotus (Nelumbo nucifera) of the Nymphaceae family embellished with MoS2 nanosheets for its enhanced hydrophobic and antimicrobial properties. MoS2 nanosheets synthesized by a coprecipitation method using sodium molybdate dihydrate (Na2MoO4·2H2O) and thioacetamide (CH3CSNH2) were used as a sourse for MoS2 particle growth with cellulose threads extracted from lotus peduncles. The size, crystallinity, and morphology of pure and MoS2-coated fibers were studied using X-ray diffractometry (XRD) and scanning electron microscopy (SEM). the XRD pattern of pure lotus threads showed a semicrystalline nature, and the threads@MoS2 composite showed more crystallinity than the pure threads. SEM depicts that pure lotus threads possess a smooth surface, and the MoS2 nanosheets growth can be easily identified on the threads@MoS2. Further, the presence of MoS2 nanosheets on threads was confirmed with EDX elemental analysis. Antimicrobial studies with Escherichia coli and Candida albicans reveal that threads@MoS2 have better resistance than its counterpart, i.e., pure threads. MoS2 sheets play a predominant role in restricting the wicking capability of the pure threads due to their enhanced hydrophobic property. The water absorbency assay denotes the absorption rate of threads@MoS2 to 80%, and threads@MoS2 shows no penetration for the observed 60 min, thus confirming its wicking restriction. The contact angle for threads@MoS2 is 128°, indicating its improved hydrophobicity.

Paper-based microfluidic devices for food adulterants: Cost-effective technological monitoring systems.

Analytical sciences have witnessed emergent techniques for efficient clinical and industrial food adulterants detection. In this review, the contributions made by the paper-based devices are highlighted for efficient and rapid detection of food adulterants and additives, which is the need of the hour and how different categories of techniques have been developed in the past decade for upgrading the performance for point-of-care testing. A simple strategy with an arrangement for detecting specific adulterants followed by the addition of samples to obtain well-defined qualitative or quantitative signals for confirming the presence of target species. The paper-based microfluidics-based technology advances and prospects for food adulterant detection are discussed given the high-demand from the food sectors and serve as a valued technology for food researchers working in interdisciplinary technological frontiers.

Paper and thread as media for the frugal detection of urinary tract infections (UTIs).

Urinary tract infections (UTIs) make up a significant proportion of the global burden of disease in vulnerable groups and tend to substantially impair the quality of life of those affected, making timely detection of UTIs a priority for public health. However, economic and societal barriers drastically reduce accessibility of traditional lab-based testing methods for critical patient groups in low-resource areas, negatively affecting their overall healthcare outcomes. As a result, cellulose-based materials such as paper and thread have garnered significant interest among researchers as substrates for so-called frugal analytical devices which leverage the material's portability and adaptability for facile and reproducible diagnoses of UTIs. Although the field may be only in its infancy, strategies aimed at commercial penetration can appreciably increase access to more healthcare options for at-risk people. In this review, we catalogue recent advances in devices that use cellulose-based materials as the primary housing or medium for UTI detection and chart out trends in the field. We also explore different modalities employed for detection, with particular emphasis on their ability to be ported onto discreet casings such as sanitary products.

"Scratch it out": carbon copy based paper devices for microbial assays and liver disease diagnosis.

We present a facile paper-based microfluidic device fabrication technique leveraging off-the-shelf carbon paper for the deposition of hydrophobic barriers using a novel "stencil scratching" method. This exceedingly frugal approach (0.05$) requires practically no technical training to employ. Hydrophobic barriers fabricated using this approach offer a width of 3 mm and a hydrophilic channel width of 849 µm, with an ability to confine major aqueous solvents without leakage. The utility of the device is demonstrated by porting a cell viability assay showing a limit-of-detection (LOD) of 0.6 × 108 CFU mL-1 and bilirubin assay with human serum showing a detection range of 1.76-6.9 mg dL-1 and a limit-of-detection (LOD) of 1.76 mg dL-1. The intuitiveness and economic viability of the fabrication method afford it great potential in the field of point-of-care diagnostics geared towards providing testing infrastructure in resource-scarce regions globally.

Knitting Thread Devices: Detecting Candida albicans Using Napkins and Tampons.

Reproducible and in situ microbial detection, particularly of microbes significant in urinary tract infections (UTIs) such as Candida albicans, provides a unique opportunity to bring equity in the healthcare outcomes of disenfranchised groups like women in low-resource settings. Here, we demonstrate a system to potentially detect vulvovaginal candidiasis by leveraging the properties of multifilament cotton threads in the form of microfluidic-thread-based analytical devices (µTADs) to develop a frugal microbial identification assay. A facile mercerization method using heptane wash to boost reagent absorption and penetration is also performed and is shown to be robust compared to other existing conventional mercerization methods. Furthermore, the twisted mercerized fibers are drop-cast with media consisting of l-proline ß-naphthylamide, which undergoes hydrolysis by the enzyme l-proline aminopeptidase secreted by C. albicans, hence signaling the presence of the pathogen via simple color change with a limit of detection of 0.58 × 106 cfu/mL. The flexible and easily disposable thread-based detection device when integrated with menstrual hygiene products showed a detection time of 10 min using spiked vaginal discharge. The developed method boasts a long shelf life and high stability, making it a discreet detection device for testing, which provides new vistas for self-testing multiple diseases that are considered taboo in certain societies.

Electro-kinetically driven route for highly sensitive blood pathology on a paper-based device.

Enhancing the sensitivity of colorimetric detection in paper-devices is a quintessential step in achieving frugal diagnosis. Here, we demonstrate an effective way of improving the detection sensitivity of paper-based devices, as mediated by electro-kinetic mechanisms. By directly employing blood plasma, we investigate the electro-kinetic clustering of glucose, a neutral molecule in paper devices. Under the influence of uniform electric field, dispersed glucose gets accumulated in the paper strips. Due to the combination of EOF and electrophoretic migration, we achieve twofold increase in the colour intensity for both normal and diabetic samples. This approach is robust and possesses better sensitivity than conventional colorimetric assays and can be easily extended to other body fluid based diagnosis. These results may turn out to be of profound importance in improving the quality of pathological diagnosis in low-cost paper-based point-of-care devices deployed in resource-limited settings.

Thread integrated smart-phone imaging facilitates early turning point colorimetric assay for microbes.

This study employs a commercial multifilament cotton thread as a low-cost microbial identification assay integrated with smartphone-based imaging for high throughput and rapid detection of pathogens. The thread device with inter-twined fibers was drop-cast with test media and a pH indicator. The target pathogens scavenge the media components with different sugars and release acidic by-products, which in turn act as markers for pH-based color change. The developed thread-based proof-of-concept was demonstrated for the visual color detection (red to yellow) of Candida albicans (≈16 hours) and Escherichia coli (≈5 hours). Besides that, using a smart-phone to capture images of the thread-based colorimetric assay facilitates early detection of turning point of the pH-based color change and further reduces the detection time of pathogens viz. Candida albicans (≈10 hours) and Escherichia coli (≈1.5 hours). The reported thread and smartphone integrated image analysis works towards identifying the turning point of the colorimetric change rather than the end-point analysis. Using this approach, the interpretation time can be significantly reduced compared to the existing conventional microbial methods (≈24 hours). The thread-based colorimetric microbial assay represents a ready-to-use, low-cost and straightforward technology with applicability in resource-constrained environments, surpassing the need for frequent fresh media preparation, expensive instrumentation, complex fabrication techniques and expert intervention. The proposed method possesses high scalability and reproducibility, which can be further extended to bio(chemical) assays.

Fabricating Paper Based Devices Using Correction Pens.

We present a rapid (<10 s), cost-effective, unique single-step method for fabricating paper-based devices without necessitating any expensive instrumentation, simply by deploying correction pens that are otherwise commonly used for masking typos in printed or written matters. The marked regions formed by deposits from the correction pen demonstrate ubiquitous flow resistances to typical aqueous solutions and organic solvents in the transverse direction, resulting in a preferential bulk flow along the axial direction of the paper channels 'fabricated' in the process. Considering the simplicity and cost-effectiveness of this platform, it is deemed to be ideal for (bio) chemical sensing and point-of-care diagnostics in resource-limited settings.

Photosensitive surfactants with various hydrophobic tail lengths for the photocontrol of genomic DNA conformation with improved efficiency.

We report the synthesis and characterisation of photosensitive cationic surfactants with various hydrophobic tail lengths. These molecules, called AzoCx, are used as photosensitive nucleic acid binders (pNABs) and are applied to the photocontrol of DNA conformation. All these molecules induce DNA compaction in a photodependent way, originating in the photodependent polarity of their hydrophobic tails. We show that increasing hydrophobicity strongly enhances the compaction efficiencies of these molecules, but reduces the possibility of reversible photocontrol of a DNA conformation. Optimal performance was achieved with AzoC5, which allowed reversible control of DNA conformation with light at a concentration seven times smaller than previously reported.