Fibrin/MoS-nanosheet conductive hydrogels with programmed time scales and pathways for bioresorption.

Electrically conductive hydrogels are of interest as scaffolds for tissue engineering applications involving the growth, implantation, or attachment of electrically active cells. Such hydrogels should exhibit soft mechanics, tunable conductivity to match native tissue, biocompatibility, and biodegradability into non-toxic, clearable species. Common conductors based on metals or polymers can be challenged by insufficient biocompatibility or biodegradability.

Characterizing ssRNA and dsRNA electrophoretic behavior: empirical insights with neural network-aided predictions.

RNA-based therapeutics are currently at the forefront of the biopharmaceutical industry because of their safety, efficacy, and shortened time from disease discovery to therapy development. Microfluidic electrophoresis provides a great analytical platform to analyze nucleic acids in unprecedented detail. However, while DNA has been studied extensively within microfluidic systems, there is limited data available for RNA, particularly of chemically modified molecules, such as those used in the COVID-19 mRNA vaccines, and for long double-stranded RNA molecules, which may accompany, intentionally or as a by-product, RNA therapeutics.

Opioid quantification via microsampling techniques to assess opioid use in human laboratory studies.

Despite progress in neurobiological studies with human subjects, sample availability remains a challenge. Urine samples, widely used for screening, suffer from false-positive results due to immunoassay cross-reactivity. Serum, used for confirmatory testing, offers advantages but faces limitations due to blood collection.

Precision through electric-field assisted automatable high throughput sample preparation of dried blood spots for neonatal abstinence syndrome detection.

In the United States, approximately 20 % of pregnant women disclose opioid misuse, contributing significantly to the widespread occurrence of Neonatal Abstinence Syndrome (NAS) in neonates exposed to opioids during gestation. Current NAS diagnosis heavily relies on clinical observation of symptoms, with the Finnegan Neonatal Abstinence Scoring System (FNASS) serving as the gold standard due to challenges associated with obtaining biological specimens from newborns. This methodological constraint poses difficulties in achieving accurate quantitative assessments and implementing timely therapeutic interventions.

Utilizing Functionalized Dual-Microbead Interfaces for Measurements of Amyloid-beta (1-42) with Femtomolar Sensitivity.

The presence of amyloid-beta (1-42) in CSF and plasma has been implicated as a pathological hallmark of Alzheimer's disease. Here, we demonstrate a high-sensitivity method of detection and quantification for total Aβ42 utilizing two-antibody functionalized microparticles interfaces. Our method of biomarker detection utilizes two types of microbeads: (1) magnetic particles functionalized with monoclonal antibodies and (2) polystyrene particles dual functionalized with monoclonal antibodies and a unique DNA "tag".

Review of microRNA detection workflows from liquid biopsy for disease diagnostics.

MicroRNAs have emerged as effective biomarkers in disease diagnostics, particularly cancer, due to their role as regulatory sequences. More recently, microRNAs have been detected in liquid biopsies, which hold immense potential for early disease diagnostics. This review comprehensively analyses distinct liquid biopsy microRNA detection methods validated with clinical samples.

BMT: A Cross-Validated ThinPrep Pap Cervical Cytology Dataset for Machine Learning Model Training and Validation.

In the past several years, a few cervical Pap smear datasets have been published for use in clinical training. However, most publicly available datasets consist of pre-segmented single cell images, contain on-image annotations that must be manually edited out, or are prepared using the conventional Pap smear method. Multicellular liquid Pap image datasets are a more accurate reflection of current cervical screening techniques.

How to convert a 3D printer to a personal automated liquid handler for life science workflows.

Automated liquid handlers are fundamental in modern life science laboratories, yet their high costs and large footprints often limit accessibility for smaller labs. This study presents an innovative approach to decentralizing a liquid handling system by converting a low-cost 3D printer into a customizable and accurate liquid handler. The Personal Automated Liquid Handler (PALH) system, costing ∼$400, incorporates a single-channel pipet, custom 3D-printed components, and open-source software for personalized workflows, allowing researchers to build and modify the system for specific experimental needs.

Peripheral blood to next-generation sequencing ready DNA library: a novel engineering design for automation.

Whole genome sequencing (WGS) has become a gold standard for diagnosing genomic variation. Peripheral blood is a common sample source for the extraction of nucleic acids for Next-Generation Sequencing (NGS) applications. Here, we present an integrated and fully automated device design that uses new concepts of fluid mechanics, heat-mass transfer, and thermodynamics of enzymatic reactions to extract nucleic acids from the blood and perform DNA library preparation from a pre-filled plate.

Leveraging the fundamentals of heat transfer and fluid mechanics in microscale geometries for automated next-generation sequencing library preparation.

Next-generation sequencing (NGS) is emerging as a powerful tool for molecular diagnostics but remains limited by cumbersome and inefficient sample preparation. We present an innovative automated NGS library preparation system with a simplified mechanical design that exploits both macro- and microfluidic properties for optimizing heat transfer, reaction kinetics, mass transfer, fluid mechanics, adsorption-desorption rates, and molecular thermodynamics. Our approach introduces a unique two-cannula cylindrical capillary system connected to a programmable syringe pump and a Peltier heating element able to execute all steps with high efficiency.

Electrophoretic Microfluidic Characterization of mRNA- and pDNA-Loaded Lipid Nanoparticles.

Lipid nanoparticles (LNPs) are clinically advanced nonviral gene delivery vehicles with a demonstrated ability to address viral, oncological, and genetic diseases. However, the further development of LNP therapies requires rapid analytical techniques to support their development and manufacturing. The method developed and described in this paper presents an approach to rapidly and accurately analyze LNPs for optimized therapeutic loading by utilizing an electrophoresis microfluidic platform to analyze the composition of LNPs with different clinical lipid compositions (Onpattro, Comirnaty, and Spikevax) and nucleic acid (plasmid DNA (pDNA) and messenger RNA (mRNA)) formulations.

Assessment of pDNA isoforms using microfluidic electrophoresis.

The recent rise in nucleic acid-based vaccines and therapies has resulted in an increased demand for plasmid DNA (pDNA). As a result, there is added pressure to streamline the manufacturing of these vectors, particularly their design and construction, which is currently considered a bottleneck. A significant challenge in optimizing pDNA production is the lack of high-throughput and rapid analytical methods to support the numerous samples produced during the iterative plasmid construction step and for batch-to-batch purity monitoring.

Microfluidic AAV Purity Characterization: New Insights into Serotype and Sample Treatment Variability.

Despite recent advances in nucleic acid delivery systems with the success of LNP vehicles, adeno-associated virus (AAV) remains the leading platform for targeted gene delivery due to its low immunogenicity to humans, high transduction efficiency, and range of serotypes with varying tropisms. Depending on the therapeutic goals and serotype used, different production conditions may be more amenable, generating an ever-growing need for rapid yet robust analytical techniques to support the high-quality manufacturing of AAV. A critical bottleneck exists for assessing full capsids where rapid, high-throughput techniques capable of analyzing a range of serotypes are needed.

Enzymatic isolation and microfluidic electrophoresis analysis of residual dsRNA impurities in mRNA vaccines and therapeutics.

The versatility, rapid development, and ease of production scalability of mRNA therapeutics have placed them at the forefront of biopharmaceutical research. However, despite their vast potential to treat diseases, their novelty comes with unsolved analytical challenges. A key challenge in ensuring sample purity has been monitoring residual, immunostimulatory dsRNA impurities generated during the transcription of mRNA.

A microfluidic electrophoretic dual dynamic staining method for the identification and relative quantitation of dsRNA contaminants in mRNA vaccines.

mRNA vaccines (, COVID-19 vaccine) offer various advantages over traditional vaccines in preventing and reducing disease and shortening the time between pathogen discovery and vaccine creation. Production of mRNA vaccines results in several nucleic acid and enzymatic by-products, most of which can be detected and removed; however, double-stranded RNA (dsRNA) contaminants pose a particular challenge. Current purification and detection platforms for dsRNA vary in effectiveness, with problems in scalability for mass mRNA vaccine production.

Insight into Increased Recovery and Simplification of Genomic DNA Extraction Methods from Dried Blood Spots.

There is no consensus on how to perform the manual extraction of nucleic acids from dried blood spots (DBSs). Current methods typically involve agitation of the DBSs in a solution for varying amounts of time with or without heat, and then purification of the eluted nucleic acids with a purification protocol. We explored several characteristics of genomic DNA (gDNA) DBS extraction such as extraction efficiency, the role of red blood cells (RBCs) in extraction and critical kinetic factors to understand if these protocols can be simplified while maintaining sufficient gDNA recovery.

An in-depth analysis of four classes of antidepressants quantification from human serum using LC-MS/MS.

Depression is a growing global crisis, with females at a higher rate of diagnosis than males. While the percentage of patients on prescribed antidepressants have tripled over the last two decades, we are still at a crossroad where the discrepancy lies between finding a drug to suit a patient and monitoring the abundance of it in the body to prevent unwanted side effects. Liquid Chromatography tandem mass spectrometry (LC-MS/MS) has garnered the attention of clinicians as a technique to accurately monitor therapeutic drugs in human serum with high specificity and accuracy.

An Optimized CoBRA Method for the Microfluidic Electrophoresis Detection of Breast Cancer Associated Methylation.

Although breast cancer screening assays exist, many are inaccessible and have high turnaround times, leaving a significant need for better alternatives. Hypermethylation of tumor suppressor genes is a common epigenetic marker of breast cancer. Methylation tends to occur most frequently in the promoter and first exon regions of genes.

Ultrasound frequency sonication facilitates high-throughput and uniform dissociation of cellular aggregates and tissues.

A sample preparation step involving dissociation of tissues into their component cells is often required to conduct analysis of nucleic acids and other constituents from tissue samples. Frequently, the extracellular matrix and cell-cell adhesions are disrupted via treatment with a chemical dissociating reagent or various mechanical forces. In this work, a new, high-throughput, multiplexed method of dissociating tissues and cellular aggregates into single cells using ultrasound frequency bath sonication is explored and characterized.

Electro-DBS: A Simple Method to Rapidly Extract Genomic DNA from Dried Blood Spots.

Dried blood spots (DBSs) have been used for more than 50 years and are used as a method of sample collection for pharmacological testing, genetic analyses, monitoring of viral infections, and more. Protocols for nucleic acid extraction from DBSs involve several steps that require specialized equipment and can be lengthy. Thus, we sought to explore ways to reduce the analytical burden of DBSs.

A Comparative Analysis of Deep Learning Models for Automated Cross-Preparation Diagnosis of Multi-Cell Liquid Pap Smear Images.

Routine Pap smears can facilitate early detection of cervical cancer and improve patient outcomes. The objective of this work is to develop an automated, clinically viable deep neural network for the multi-class Bethesda System diagnosis of multi-cell images in Liquid Pap smear samples. 8 deep learning models were trained on a publicly available multi-class SurePath preparation dataset.

An investigation into simplifying total RNA extraction with minimal equipment using a low volume, electrokinetically driven microfluidic protocol.

Current methods for total RNA extraction are time-consuming and require several hands-on steps and specialized equipment. Microfluidic devices can offer the opportunity to reduce the number of hands-on steps, decrease the volumes of reagents required for purification, and make extraction high throughput. Here, we investigated the translation of a high volume magnetic bead-based total RNA extraction method (from human whole blood) onto a low input volume microfluidic device.

Electrophoresis-Mediated Characterization of Full and Empty Adeno-Associated Virus Capsids.

Adeno-associated virus (AAV) has shown great potential in gene therapy due to its low immunogenicity, lack of pathogenicity to humans, and ability to provide long-term gene expression . However, there is currently a need for fast, high-throughput characterization systems that require low volumes for the determination of its sample composition in terms of full and empty capsids since empty capsids are a natural byproduct of AAV synthesis. To address this need, the following study proposes a high-throughput electrophoresis-mediated microfluidics approach that is independent of sample input concentration to estimate the composition of a given sample by combining its protein and ssDNA information relative to a standard.

Electric-field facilitated rapid and efficient dissociation of tissues Into viable single cells.

Single-Cell Analysis is a growing field that endeavors to obtain genetic profiles of individual cells. Disruption of cell-cell junctions and digestion of extracellular matrix in tissues requires tissue-specific mechanical and chemical dissociation protocols. Here, a new approach for dissociating tissues into constituent cells is described.

Pre-eclampsia: a Scoping Review of Risk Factors and Suggestions for Future Research Direction.

Abstract: Most of maternal deaths are preventable, and one-quarter of maternal deaths are due to pre-eclampsia and eclampsia. Prenatal screening is essential for detecting and managing pre-eclampsia. However, pre-eclampsia screening is solely based on maternal risk factors and has low (< 5% in the USA) detection rates.