A miniaturized additive-manufactured carbon black/PLA electrochemical sensor for pharmaceuticals detection.

Additive manufacturing is a technique that allows the construction of prototypes and has evolved a lot in the last 20 years, innovating industrial fabrication processes in several areas. In chemistry, additive manufacturing has been used in several functionalities, such as microfluidic analytical devices, energy storage devices, and electrochemical sensors. Theophylline and paracetamol are important pharmaceutical drugs where overdosing can cause adverse effects, such as tachycardia, seizures, and even renal failure. Therefore, this paper aims at the development of miniaturized electrochemical sensors using 3D printing and polylactic acid-based conductive carbon black commercial filament for theophylline and paracetamol detection. Electrochemical characterizations of the proposed sensor were performed to prove the functionality of the device. Morphological characterizations were carried out, in which chemical treatment could change the surface structure, causing the improvement of the analytical signal. Thus, the detection of theophylline at a linear range of 5.00-150 µmol L-1 with a limit of detection of 1.2 µmol L-1 was attained, and the detection of paracetamol at a linear range of 1.00-200 µmol L-1 with a limit of detection of 0.370 µmol L-1 was obtained, demonstrating the proposed sensor effectively detected pharmaceutical drugs.

Miniaturized flexible micro-tube plasma ionization source for the effective ionization of non-easily ionizable pesticides in food with liquid chromatography/mass spectrometry.

In this article, we have studied the potential of flexible microtube plasma (FµTP) as ionization source for the liquid chromatography high-resolution mass spectrometry detection of non-easily ionizable pesticides (viz. nonpolar and non-ionizable by acid/basic moieties). Phthalimide-related compounds such as dicofol, dinocap, o-phenylphenol, captan, captafol, folpet and their metabolites were studied. Dielectric barrier discharge ionization (DBDI) was examined using two electrode configurations, including the miniaturized one based on a single high-voltage (HV) electrode and a virtual ground electrode configuration (FµTP), and also the two-ring electrode DBDI configuration. Different ionization pathways were observed to ionize these challenging, non-easily ionizable nonpolar compounds, involving nucleophilic substitutions and proton abstraction, with subtle differences in the spectra obtained compared with APCI. An average sensitivity increase of 5-fold was attained compared with the standard APCI source. In addition, more tolerance with matrix effects was observed in both DBDI sources. The importance of the data reported is not just limited to the sensitivity enhancement compared to APCI, but, more notably, to the ability to effectively ionize nonpolar, late-eluting (in reverse-phase chromatography) non-ionizable compounds. Besides o-phenylphenol ([M - H]-), all the parent species were efficiently ionized through different mechanisms involving bond cleavages through the effect of plasma reagent species or its combination with thermal degradation and subsequent ionization. This tool can be used to figure out overlooked nonpolar compounds in different environmental samples of societal interest through non-target screening (NTS) strategies.

The Use of the RETeval Portable Electroretinography Device for Low-Cost Screening: A Mini-Review.

Electroretinography (ERG) provides crucial insights into retinal function and the integrity of the visual pathways. However, ERG assessments classically require a complicated technical background with costly equipment. In addition, the placement of corneal or conjunctival electrodes is not always tolerated by the patients, which restricts the measurement for pediatric evaluations. In this short review, we give an overview of the use of the RETeval portable ERG device (LKC Technologies, Inc., Gaithersburg, MD, USA), a modern portable ERG device that can facilitate screening for diseases involving the retina and the optic nerve. We also review its potential to provide ocular biomarkers in systemic pathologies, such as Alzheimer's disease and central nervous system alterations, within the framework of oculomics.

Miniaturization of an enclosed electrospinning process to enhance reproducibility in the fabrication of rapidly dissolving cell-based biosensors.

There is broad interest in producing electrospun films embedded with biological materials. It is well known that electrospinning requires careful control of the process conditions, especially the environmental conditions such as relative humidity (RH). Given that commercial electrospinning systems are expensive (> $10,000) and are typically too large to be used in standard biological safety cabinets (BSC), we designed and built a miniaturized electrospinning box (E-Box) that will fit inside a BSC, and the RH can be easily controlled using simple instrumentation (gas cylinder, regulator, needle valve, rotameter). It uses an inexpensive computerized numerical control machine to control the spinneret positioning and collector rotational speed-all the parts for the device (except the syringe pump and voltage supply) can be purchased for approximately $1000. We demonstrate the usefulness of our design in optimizing the production of Escherichia coli-embedded pullulan-trehalose films to be used as rapidly dissolving biosensors for environmental monitoring. At a fixed electrospinning recipe, we showed that decreasing the RH from approximately 48% to 22% resulted in the average fiber diameter increasing from 240 (± 11) nm to 314 (± 8) nm. We also demonstrate the usefulness of our design in performing sequential electrospinning experiments to evaluate process performance reproducibility. For example, from just 1 mL of a polymer solution, we produced 16 electrospun films (approximately 3 cm by 8 cm each)-from those films we hole-punched approximately 80 biosensor discs which were then used in subsequent experiments to determine the amount of two different biocides (Grotan BK and triclosan) in aqueous samples. The technique developed in this study is ideal for creating electrospun materials in high quantities that are highly reproducible through the precise control of RH.

A strategy for Cas13 miniaturization based on the structure and AlphaFold.

The small size of the Cas nuclease fused with various effector domains enables a broad range of function. Although there are several ways of reducing the size of the Cas nuclease complex, no efficient or generalizable method has been demonstrated to achieve protein miniaturization. In this study, we establish an Interaction, Dynamics and Conservation (IDC) strategy for protein miniaturization and generate five compact variants of Cas13 with full RNA binding and cleavage activity comparable the wild-type enzymes based on a combination of IDC strategy and AlphaFold2. In addition, we construct an RNA base editor, mini-Vx, and a single AAV (adeno-associated virus) carrying a mini-RfxCas13d and crRNA expression cassette, which individually shows efficient conversion rate and RNA-knockdown activity. In summary, these findings highlight a feasible strategy for generating downsized CRISPR/Cas13 systems based on structure predicted by AlphaFold2, enabling targeted degradation of RNAs and RNA editing for basic research and therapeutic applications.

Submillimeter Multifunctional Ferromagnetic Fiber Robots for Navigation, Sensing, and Modulation.

Small-scale robots capable of remote active steering and navigation offer great potential for biomedical applications. However, the current design and manufacturing procedure impede their miniaturization and integration of various diagnostic and therapeutic functionalities. Herein, submillimeter fiber robots that can integrate navigation, sensing, and modulation functions are presented. These fiber robots are fabricated through a scalable thermal drawing process at a speed of 4 meters per minute, which enables the integration of ferromagnetic, electrical, optical, and microfluidic composite with an overall diameter of as small as 250 µm and a length of as long as 150 m. The fiber tip deflection angle can reach up to 54o under a uniform magnetic field of 45 mT. These fiber robots can navigate through complex and constrained environments, such as artificial vessels and brain phantoms. Moreover, Langendorff mouse hearts model, glioblastoma micro platforms, and in vivo mouse models are utilized to demonstrate the capabilities of sensing electrophysiology signals and performing a localized treatment. Additionally, it is demonstrated that the fiber robots can serve as endoscopes with embedded waveguides. These fiber robots provide a versatile platform for targeted multimodal detection and treatment at hard-to-reach locations in a minimally invasive and remotely controllable manner.

Microfluidic based single cell or droplet manipulation: Methods and applications.

The isolation of single cell or droplet is first and crucial step to single-cell analysis, which is important for cancer research and diagnostic methods. This review provides an overview of technologies that are currently used or in development to realize the isolation. Microfluidic based manipulation is an emerging technology with the distinct advantages of miniaturization and low cost. Therefore, recent developments in microfluidic isolated methods have attracted extensive attention. We introduced herein five strategies based on microfluid: trap, microfluidic discrete manipulation, bioprinter, capillary and inertial force. For every technology, their basic principles and features were discussed firstly. Then some modified approaches and applications were listed as the extension. Finally, we compared the advantages and drawbacks of these methods, and analyzed the trend of the manipulation based on microfluidics.

Flexible Needle Steering with Tethered and Untethered Actuation: Current States, Targeting Errors, Challenges and Opportunities.

Accurate needle targeting is critical for many clinical procedures, such as transcutaneous biopsy or radiofrequency ablation of tumors. However, targeting errors may arise, limiting the widespread adoption of these procedures. Advances in flexible needle (FN) steering are emerging to mitigate these errors. This review summarizes the state-of-the-art developments of FNs and addresses possible targeting errors that can be overcome with steering actuation techniques. FN steering techniques can be classified as passive and active. Passive steering directly results from the needle-tissue interaction forces, whereas active steering requires additional forces to be applied at the needle tip, which enhances needle steerability. Therefore, the corresponding targeting errors of most passive FNs and active FNs are between 1 and 2 mm, and less than 1 mm, respectively. However, the diameters of active FNs range from 1.42 to 12 mm, which is larger than the passive steering needle varying from 0.5 to 1.4 mm. Therefore, the development of active FNs is an area of active research. These active FNs can be steered using tethered internal direct actuation or untethered external actuation. Examples of tethered internal direct actuation include tendon-driven, longitudinal segment transmission and concentric tube transmission. Tendon-driven FNs have various structures, and longitudinal segment transmission needles could be adapted to reduce tissue damage. Additionally, concentric tube needles have immediate advantages and clinical applications in natural orifice surgery. Magnetic actuation enables active FN steering with untethered external actuation and facilitates miniaturization. The challenges faced in the fabrication, sensing, and actuation methods of FN are analyzed. Finally, bio-inspired FNs may offer solutions to address the challenges faced in FN active steering mechanisms.

Gyrolab Immunoassays: Miniaturization, Automation, and Integration into a Rapid Workflow.

Gyrolab® is an open immunoassay platform that automates the complete immunoassay protocol in a microfluidic disc. The column profiles generated with Gyrolab immunoassays are used to gain more information about biomolecular interactions that can be useful in assay development or quantify analytes in samples. Gyrolab immunoassays can be used to cover a broad concentration range and diversity of matrices in applications ranging from biomarker monitoring, pharmacodynamics and pharmacokinetics studies, to bioprocess development in many areas, including therapeutic antibodies, vaccines, and cell and gene therapy.This chapter is an overview of Gyrolab technology, including system components and the assay development workflow, including the process of selecting affinity reagents, Gyrolab Bioaffy CDs, and assay conditions to optimize immunoassays. Two case studies are included. The first involves an assay for the humanized antibody pembrolizumab used in cancer immunotherapy that can generate data for pharmacokinetics studies. The second case study involves quantification of the biomarker and biotherapeutic interleukin-2 (IL-2) in human serum and buffer. IL-2 has been implicated in the cytokine storm associated with COVID-19, and cytokine release syndrome (CRS), which can occur during chimeric antigen receptor T cell (CART) therapy used in treating cancer. These molecules also have therapeutic relevance in combination.

Trends in monoliths: Packings, stationary phases and nanoparticles.

Monoliths media are gaining interest as excellent substitutes to conventional particle-packed columns. Monolithic columns show higher permeability and lower flow resistance than conventional liquid chromatography columns, providing high-throughput performance, resolution and separation in short run times. Monolithic columns with longer length, smaller inner diameter and specific selectivity to peptides or enantiomers have been played important role in hyphenated system. Monolithic stationary phases possess great efficiency, resolution, selectivity and sensitivity in the separation of complex biological samples, such as the complex mixtures of peptides for proteome analysis. The development of monolithic stationary phases has opened the new avenue in chromatographic separation science and is in turn playing much more important roles in the wide application area. Monolithic stationary phases have been widely used in fast and high efficiency one- and multi-dimensional separation systems, miniaturized devices, and hyphenated system coupled with mass spectrometers. The developing technology for preparation of monolithic stationary phases is revolutionizing the column technology for the separation of complex biological samples. These techniques using porous monoliths offer several advantages, including miniaturization and on-line coupling with analytical instruments. Additionally, monoliths are ideal support media for imprinting template-specific sites, resulting in the so-called molecularly-imprinted monoliths, with ultra-high selectivity. In this review, the origin of the concept, the differences between their characteristics and those of traditional packings, their advantages and drawbacks, theory of separations, the methods for the monoliths preparation of different forms, nanoparticle monoliths and metal-organic framework are discussed. Two application areas of monolithic metal-organic framework and nanoparticle monoliths are provided. The review article discusses the results reported in a total of 218 references. Other older references were included to illustrate the historical development of monoliths, both in preparation and types, as well as separation mechanism.

Robotic probes at the cell scale.

The miniaturization of robotic tools and probes enables the fundamental study of mechanical properties of cells and tissues.

Quality and utility of YouTube videos about mPCNL.

This study assessed the reliability, efficacy, and quality of YouTube videos on miniaturized PCNL procedures using a scoring system. Video searches were conducted using the keywords miniaturized percutaneous nephrolithotomy (mPCNL), ultra-mini-PCNL (UMP), and super-mini-PCNL (SMP) via https://www.youtube.com . All videos > 2 min in English containing the procedure of mPCNL were ordered and listed according to relevance and popularity. Global quality scores (GQS) and reliability scales were used for quality and content analysis of the videos. The confirmed video power index (VPI) was used to assess video popularity, and the mPCNL score, developed by three endourologists, was used to measure the informational and educational value of the videos. While specialists (61.9%) were the main source for uploading 63 videos included in the study, the largest target group was patients (61.9%). Operational procedures (68.3%) were the main content, and the highest upload region was Asian continent (66.7%). The majority of videos were in English (60.3%), focused on standard mPCNL rather than miniaturization subtypes (77.8%). The mean VPI, GQS, reliability, and mPCNL scores were 4.61, 2.86, 2.61, 9.58, respectively. These parameters, which were positively correlated, were found to be statistically significantly higher for videos of 'academic' origin, 'Physicians' target audience and 'English audio' language format (p < 0.05). Because mPCNL videos, which are popular and have high informational and educational value, are aimed at medical professionals rather than patients, they are not functional in treatment selection and explaining the procedure. By raising its standards, video portals can make the treatment process practical as a functional bridge between patients and health care professionals.

Miniaturized systems for gas chromatography: Developments in sample preparation and instrumentation.

An important field of research is the miniaturization of analytical systems for laboratory applications and on-field analysis. In particular, gas chromatography (GC) has benefited from the recent advances in enabling technologies like photolithography, micromachining, hot embossing, and 3D-printing to improve sampling and sample preparation, microcolumn technologies, and detection. In this article, the developments and applications reported since 2015 were reviewed and summarized. Important applications using benchtop instruments, portable GCs, and micro-GCs (µGCs) were showcased to illustrate the current challenges associated with each miniaturized interfaces and systems. For instance, portable instruments need to be energy-efficient and ideally depend on renewable sources for carrier gas generation. Lastly, multidimensional separations were addressed using miniaturized systems to effectively improve the peak capacity of portable systems.

Small, smaller, smallest: Miniaturization of chromatographic process development.

Biopharmaceuticals are becoming increasingly important in modern healthcare. Monoclonal antibodies (mAb) are one of the most widely used therapeutic proteins and are important for the treatment of cancer and autoimmune diseases, among others. After cell culture there are still large amounts of other impurities (e.g. host cell proteins) in solution. Chromatography is usually the first purification step, allowing to increase purity and reduce volume. This comes associated with high costs and chromatography accounts for a significant portion of total production costs for therapeutic proteins. Chromatographic process development may be time consuming and use large amounts of resins. Therefore, there is increased interest in finding cheaper techniques for chromatographic process development without compromising accuracy. This paper presents a highly sophisticated microfluidic chip approach for efficient adsorption isotherm determinations compared to current chromatographic process development. Implementation of an image analysis software ensures that chromatographic resin volume is accurately determined. The adsorption isotherm performance of microfluidics was compared to the robotic Liquid-handling Station (LHS) and labor intensive Eppendorf tubes. The microfluidic chip allows a 15-fold volume reduction and resin consumptions as low as 100/200 nl (200/100-fold reduction). The microfluidic chip performed comparably to the other miniaturized techniques, using less liquid and resin volume. For process development of expensive products (e.g. monoclonal antibodies), miniaturization (provided by the microfluidic chip) proved to be the most cost effective alternative whereas for less valuable products (e.g. lysozyme) automation (provided by the LHS) was the most cost effective alternative.

Evolution of Biotelemetry in Medical Devices: From Radio Pills to mm-Scale Implants.

The advent of semiconductor technology in the mid-20th century created unprecedented opportunities to develop a new generation of small-scale wireless medical sensing devices that can support remote monitoring of patients' vital signs. The first radio pills were developed as early as the 1950's using only a few transistors. These swallowable capsules could sense and wirelessly transmit vital parameters from inside the human body. Since then we have witnessed the rapid progress of medical devices driven by the evolution of semiconductor technology, from single-transistor oscillators to complex mixed-signal multi-channel and multi-modal systems. This paper retraces the evolution of biotelemetry devices from their very early inception to the smart miniaturized systems of modern days, focusing on semiconductor-enabled sensing methods and circuits developed over the last six decades. The paper also includes the author's perspective on current and future trends in the development of CMOS-based biotelemeters, focusing on concepts of implant modularity, miniaturization and hybrid energy harvesting solutions.

A miniature robotic steerable endoscope for maxillary sinus surgery called PliENT.

In endoscopic maxillary sinus surgery, the maxillary sinus is accessed through the nasal cavity which constitutes a narrow and tortuous pathway. However, surgeons still use rigid endoscopes and rigid, straight or pre-bent instruments for this procedure. Resection of the uncinate process and creation of a medial antrostomy is warranted to access the pathology inside the maxillary sinus and depending on the location of the pathology (lateral, inferior or anterior wall), additional resection of healthy tissue and/or functional structures like the lacrimal duct and/or inferior turbinate is necessary to gain optimal access. In order to avoid this additional resection, a functional single-handed, steerable endoscope for endoscopic maxillary sinus surgery has been designed and built. This endoscope is, to our knowledge, the most slender active steerable endoscope ever reported for maxillary sinus surgery. The performance of the endoscope was validated by two surgeons on a cadaver. An increased field of view was found in comparison to currently used endoscopes. As a direct consequence, a reduced need for resection of healthy tissue was confirmed.

Hair shaft miniaturization causes stem cell depletion through mechanosensory signals mediated by a Piezo1-calcium-TNF-α axis.

In aging, androgenic alopecia, and genetic hypotrichosis disorders, hair shaft miniaturization is often associated with hair follicle stem cell (HFSC) loss. However, the mechanism causing this stem cell depletion in vivo remains elusive. Here we show that hair shaft loss or a reduction in diameter shrinks the physical niche size, which results in mechanical compression of HFSCs and their apoptotic loss. Mechanistically, cell compression activates the mechanosensitive channel Piezo1, which triggers calcium influx. This confers tumor necrosis factor alpha (TNF-α) sensitivity in a hair-cycle-dependent manner in otherwise resistant HFSCs and induces ectopic apoptosis. Persistent hair shaft miniaturization during aging and genetic hypotrichosis disorders causes long-term HFSC loss by inducing continuous ectopic apoptosis through Piezo1. Our results identify an unconventional role of the inert hair shaft structure as a functional niche component governing HFSC survival and reveal a mechanosensory axis that regulates physical-niche-atrophy-induced stem cell depletion in vivo.

Surgical Technique of Supine Percutaneous Nephrolithotomy in Children / Técnica quirúrgica de la nefrolitotomía percutánea en supino en niños

Percutaneous nephrolithotomy (PCNL) in children has becomemore widely used due to its high efficacy and safety and to the development of miniaturized instruments. A supine approach is promising due to advantages such as better ventilation, reproducibility, and ergonomics. The purpose of the present study is to describe our surgical technique with special considerations in the pediatric population. We used an oblique supine position supported by one silicone gel positioning pad under the hip and another under the ipsilateral flank. The anatomical landmarks used to guide the puncture were the 11th and 12th ribs, the posterior axillary line, and the iliac crest. Initially, a ureteral catheter was introduced endoscopically. A retrograde pyelography was performed to guide the puncture, which was performed using a biplanar technique. A hydrophilic guide wire was then advanced through the needle. Dilation was performed with Alken telescopic dilators until 14 Ch. Fragmentation was performed either with a 13 Ch semirigid cystoscope or a flexible ureteroscope using a holmium: yttrium aluminum garnet (Ho:Yag) laser.We left a double J catheter. Supine PCNL in the pediatric population has comparable efficacy in terms of stone free rate to that of the prone approach as well as less complications. Certain considerations in children are careful padding and placement of the patient close to the edge of the table. Puncture should be guided by ultrasound to reduce radiation exposure. Miniaturized equipment is not widely available, so adaptation of adult equipment for the pediatric population is sometimes necessary.

La nefrolitotomía percutánea en niños se ha vuelto ampliamente utilizada por su alta efectividad, seguridad, y por la miniaturización de los instrumentos endoscópicos. El abordaje en supino es prometedor por sus ventajas, como mejor ventilación, reproducibilidad, y ergonomía. El propósito es describir nuestra técnica quirúrgica con las consideraciones especiales a tener en cuenta en la población pediátrica. Todos nuestros pacientes han sido intervenidos bajo la siguiente técnica quirúrgica: en una posición oblicua en supino, utilizando soportes de silicona ubicados debajo de la cadera y del flanco ipsilateral, se marcan los reparos anatómicos: las costillas once y doce, la línea axilar posterior y la cresta ilíaca. Inicialmente se introduce un cateter ureteral por vía endoscópica, con el cual se realiza una pielografía retrógrada para guiar la punción con una técnica biplanar. Se avanza una guía hidrofílica y, sobre esta, los dilatadores telescopados de Alken hasta un tracto de 14 Ch. Se realiza la fragmentación con un cistoscopio semirígido de 13 Ch o con un ureteroscopio flexible utilizando el láser Ho:Yag. Se deja un cateter JJ. La nefrolitotomía percutánea en la población pediátrica es comparable en términos de tasa libre de cálculos al abordaje en prono, con menos complicaciones. Una consideración importante en niños es una adecuada posición, cerca al eje de la mesa. La punción debe ser guiada por ultrasonido para disminuir la exposición a radiación. La disponibilidad de equipos miniaturizados es limitada, por lo cual usualmente es necesario adaptar los equipos de adultos.

The influence of antenna gain and beamwidth used in OSSEUS in the screening process for osteoporosis.

Applications on electromagnetic waves in the field of biotelemetry have increased in the latest years, being used to prevent, diagnose, and treatment of several diseases. In this context, biotelemetry allows minimally invasive monitoring of the physiologic, improving comfort and patient care and significantly reducing hospital costs. Aiming to assist the mineral bone density classification, through a radio frequency signal (RF), for a later diagnosis of osteoporosis, Osseus was proposed in 2018. This equipment is a combination of the application of techniques and concepts of several areas such as software, electrical, electronic, computational, and biomedical engineering, developed at a low cost, with easy access to the population, and non-invasive. However, when placed on evaluation, potential improvements were identified to increase the stability of Osseus operation. It is proposed the implementation of improvements in the antennas used by Osseus, aiming its miniaturization, improvement in the reception of the RF signal, and better stability of the equipment's operation. Then, two antennas were built, one of which was used as a project for the second, which is an array. The array showed significant improvements in the radiation parameters relevant to the application, being a candidate to replace the antennas currently in use at Osseus.

Pushing the detection limits: strategies towards highly sensitive optical-based protein detection.

Proteins are one of the main constituents of living cells. Studying the quantities of proteins under physiological and pathological conditions can give valuable insights into health status, since proteins are the functional molecules of life. To be able to detect and quantify low-abundance proteins in biofluids for applications such as early disease diagnostics, sensitive analytical techniques are desired. An example of this application is using proteins as biomarkers for detecting cancer or neurological diseases, which can provide early, lifesaving diagnoses. However, conventional methods for protein detection such as ELISA, mass spectrometry, and western blotting cannot offer enough sensitivity for certain applications. Recent advances in optical-based micro- and nano-biosensors have demonstrated promising results to detect proteins at low quantities down to the single-molecule level, shining lights on their capacities for ultrasensitive disease diagnosis and rare protein detection. However, to date, there is a lack of review articles synthesizing and comparing various optical micro- and nano-sensing methods of enhancing the limits of detections of the antibody-based protein assays. The purpose of this article is to critically review different strategies of improving assay sensitivity using miniaturized biosensors, such as assay miniaturization, improving antibody binding capacity, sample purification, and signal amplification. The pros and cons of different methods are compared, and the future perspectives of this research field are discussed.