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Background: Phospholipids (PLs) are major constituents of cell membranes, play important roles in cell proliferation and death, as well as in signal transduction, and therefore are relevant biomarkers for different pathologies. On the other hand, when the analysis of small compounds, such as therapeutics in blood is desired, then phospholipids are part of the matrix and cause serious interference during analysis. Currently, both the analysis and removal of PLs from biological samples are limited by extensive sample preparation and instrumental separation., Results: A fast and simple quantitative Ti 4+ -modified paper spray tandem mass spectrometric (TiPS-MS/MS) method was established in urine, where the enrichment of phospholipids was achieved, as well as reduction of matrix effects (primarily caused by high salt content) that ultimately led to improved sensitivity and selectivity. The method could achieve a physiologically relevant limit of detection (0.01-0.03 μg mL -1 ). Also, the usefulness of the Ti 4+ -modified paper was investigated in the opposite mode, namely for the selective removal of phospholipids from matrices such as plasma. Clonidine is used as model compound, as the detection of this compound is known to suffer from ion suppression by phospholipids. Compared with blank paper spray tandem mass spectrometry, the limit of detection could be improved from 0.3 μg mL -1 to 0.03 μg mL -1 by employing a Ti 4+ -modified paper on top of the spray tip to capture phospholipids from the sample., Significance and Novelty: A novel Ti 4+ -modified paper was developed to allow for rapid solid-phase extraction of phospholipids from urine or selective removal from plasma, followed by direct paper spray mass spectrometric detection as a fast and convenient sample preparation and analysis combination. The paper properties are based on the Ti 4+ metal ion, which can selectively bind phosphate-containing compounds under acidic conditions, and its applicability was demonstrated in relevant biological matrices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Anti-obesity drugs, used to suppress appetite and reduce fat absorption, have been circulated and traded illegally worldwide. The traditional methods of liquid chromatography tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) for analyzing these drugs in in vivo samples require complex sample pretreatment and time-consuming procedures. To address this issue, a thermal-assisted paper spray mass spectrometry (PS-MS) method was developed in this study to analyze anti-obesity drugs in raw urine. By incorporating a heat source and optimizing the spray solvent and paper substrate, this technique demonstrates reduced matrix effect and higher sensitivity compared to traditional PS-MS methodology for direct analysis of anti-obesity drugs in urine samples. A temperature range of 100-200 °C can be set for screening anti-obesity drugs in urine samples, with the flexibility to adjust the temperature according to the specific drug being analyzed. The limits of detection (LODs) for these 15 anti-obesity drugs in urine ranged between 1 and 500 ng mL -1 . Furthermore, the thermal-assisted PS-MS method exhibited good linearities ( R 2 , 0.9903-0.9997) within the range from 10-100 to 1000 ng mL -1 for the direct quantitation of anti-obesity drugs in urine samples with an internal standard. Therefore, the thermal-assisted PS-MS technique may provide a novel approach for the direct analysis of drugs in complex samples.

Current procedures for the assessment of chronic wound infection are time-consuming and require complex instruments and trained personnel. The incidence of chronic wounds worldwide, and the associated economic burden, urge for simple and cheap point-of-care testing (PoCT) devices for fast on-site diagnosis to enable appropriate early treatment. The enzyme myeloperoxidase (MPO), whose activity in infected wounds is about ten times higher than in non-infected wounds, appears to be a suitable biomarker for wound infection diagnosis. Herein, we develop a single-component foldable paper-based device for the detection of MPO in wound fluids. The analyte detection is achieved in two steps: (i) selective immunocapture of MPO, and (ii) reaction of a specific dye with the captured MPO, yielding a purple color with increasing intensity as a function of the MPO activity in infected wounds in the range of 20-85 U/mL. Ex vivo experiments with wound fluids validated the analytic efficiency of the paper-based device, and the results strongly correlate with a spectrophotometric assay.

The prevalence of fentanyl abuse raises global public health concerns with an unprecedented surge in overdose deaths. Rapid identification and quantification of fentanyl in biofluids is of paramount importance to combat fentanyl abuse for law enforcement agencies and promptly treat patients for medical professionals. Herein, a freestanding surface-enhanced Raman spectroscopy (SERS) biosensor with excellent condensing enrichment capability, termed FrEnSERS biosensor, is reported for quantitative label-free detection of trace fentanyl in biofluids. This biosensor comprises a reduced graphene oxide membrane decorated with high-density hydrophobic Au nanostars. A combination of the high SERS enhancement and the focusing effect for analyte enrichment of the hydrophobic surface accounts for the remarkable SERS performance of the FrEnSERS biosensor. We demonstrate that the FrEnSERS biosensor achieves the sensitive and quantitative detection of fentanyl in both serum and urine over a wide dynamic range spanning more than 4 orders of magnitude, with a limit of detection of 0.47 ng/mL for serum samples and 0.73 ng/mL for urine samples. Our biosensor is sensitive, cost-effective, and reliable for rapid quantitative analysis of fentanyl in biofluids with great promise for forensic analysis and clinical diagnosis.

Three-dimensional (3D) dried blood spheroids formed on hydrophobic paper are a new microsampling platform that can stabilize labile molecules in whole blood stored in ambient air at room temperature. In this study, we define the ideal conditions for preparing the dried blood spheroids. The physical morphology of 3D dried blood spheroids is found to be largely impacted by the unregulated relative humidity of the surrounding environment. A solution of KOH placed in an enclosed chamber offers a facile way to control humidity. We also report a general polymer coating strategy that serves to stabilize dried biofluids when prepared under ordinary ambient conditions without regulation of humidity. Dried blood spheroids coated in xanthan gum polymer exhibited enhanced chemical and physical stability. The same xanthan gum polymer provided chemical stability for 2D dried blood spots when compared with the conventional noncoated samples. We have expanded the application of xanthan gum to less viscous biofluids such as urine to induce an artificial protective barrier that also provides enhanced stability for labile performance-enhancing drugs stored at room temperature. The impact of polymer coating on direct biofluid analysis via paper spray mass spectrometry was determined by comparing the relative ionization efficiency, percent difference of ionization efficiency, and matrix effects of performance-enhancing drugs that were spiked in undiluted raw urine. Acceptable analytical performance was recorded for all three criteria, including high ionization efficiencies that ranged from 77 to 93% in the presence of the xanthan gum polymer.

Chloride (Cl - ) is an inorganic anion present in a broad range of samples (e.g. biological, environmental, food, water, etc.), the determination of which is of widespread significance. In this work, we translate the well-established traditional argentometric method (Mohr's precipitation titration) into a small, simple, portable, and low-cost paper-based microfluidic diagnostic device, which provides rapid and quantitative analysis. The developed device enables the determination of chloride sample volumes as small as 5 μL. A distance-based detection method is implemented providing fully instrument-free quantitation. The beneficial effects of channel geometry (variable widths with constant heights) on analytical parameters were investigated. Trapezoidal channels (channel width changes linearly with height) were used to create a gradient of paper surface (titrant) available for the reaction, compared to the typical uniform rectangular channels (constant channel width). The trapezoid with increasing width offered higher sensitivity and lower detection limits (i.e. 0.05 mM vs 0.1 mM from the rectangular channel) for chloride determination across the concentration range of 0.05-25 mM. In addition, the effect of concentration of the deposited reagent on the obtained distance signals was investigated using varying concentrations of titrant (AgNO 3 ), which allowed determination of chloride across a wider dynamic range (up to 200 mM). The utility of the paper devices was demonstrated by determination of chloride in a variety of matrices including body fluids (sweat, serum, and urine) and water samples (drinking, mineral, river water)., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Recently, the phenomenon of fentanyls overdose leading to death is emerging in an endless stream. There is an urgent requirement to quickly identify fentanyl content in body fluids for medical and judicial purposes. With this in mind, we present a paper-based SERS substrate decorated with uniform gold nanospheres treated by chloride ion for the detection of fentanyl citrate in urine and serum. In particular, the paper-based SERS sensor was prepared by liquid/liquid self-assembly technique and chloride ion was introduced to clean and modify the substrate surface, which improved the sensitivity of the solid substrate with an enhancement factor (EF) as high as 1.64 × 10 5 . Moreover, the uniformity of each paper-based substrate and the repeatability on different batches of substrate were excellent, and there was no obvious change in the intensity response of Raman spectra within a month. As a result, the quantitative analysis of fentanyl citrate in artificial urine and rat serum were performed based on the modified paper-based substrate with the limit of detection as low as 0.59 μg/mL and 2.78 μg/mL, respectively. Both the concentrations of the two biological samples with the Raman signal intensity were linearly plotted and the recovery of the spiked samples with different concentrations was collected to verify the accuracy of the quantitative curves. All the results suggest that this work makes SERS method available for the rapid identification and quantitative analysis of illicit drug in the real biological samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Paper spray MS is part of a cohort of ambient ionization or direct analysis methods that seek to analyze complex samples without prior sample preparation. Extraction and electrospray ionization occur directly from the paper substrate upon which a dried matrix spot is stored. Paper spray MS is capable of detecting drugs directly from dried blood, plasma and urine spots at the low ng/ml to pg/ml levels without sample preparation. No front end separation is performed, so MS/MS or high-resolution MS is required. Here, we discuss paper spray methodology, give a comprehensive literature review of the use of paper spray MS for bioanalysis, discuss technological advancements and variations on this technique and discuss some of its limitations.

Ambient electrostatic paper spray ionization from a hydrophobic paper occurs when a DC potential is applied to the dry paper triangle. Online liquid/liquid extraction of small organic compounds from a drop of biological fluid present on the dry hydrophobic paper is achieved with an organic spray solvent in under 1 min and utilizes in situ electrostatic-spray ionization for more efficient detection of extracted molecules. Direct analysis of small volumes of biofluids with no sample pretreatment is possible, which is applicable in point-of-care analyses. High sensitivity and quantitative accuracy was achieved for the direct analysis of illicit drugs in 4 μL of raw blood, serum, and whole urine. The study was extended to monitor the activity of alanine transaminase enzyme, a key biomarker for the detection of liver injury in patients (with HIV and tuberculosis) who typically take several medications at once.

We report the first demonstration of using heat on a paper device to rapidly concentrate a clinically relevant analyte of interest from a biological fluid. Our technology relies on the application of localized heat to a paper strip to evaporate off hundreds of microliters of liquid to concentrate the target analyte. This method can be used to enrich for a target analyte that is present at low concentrations within a biological fluid to enhance the sensitivity of downstream detection methods. We demonstrate our method by concentrating the tuberculosis-specific glycolipid, lipoarabinomannan (LAM), a promising urinary biomarker for the detection and diagnosis of tuberculosis. We show that the heat does not compromise the subsequent immunodetectability of LAM, and in 20 min, the tuberculosis biomarker was concentrated by nearly 20-fold in simulated urine. Our method requires only 500 mW of power, and sample flow is self-driven via capillary action. As such, our technology can be readily integrated into portable, battery-powered, instrument-free diagnostic devices intended for use in low-resource settings.

In recent years, calcium phosphate-base composites, such as hydroxyapatite (HA) and carbonate apatite (CA) have been considered desirable and biocompatible coating layers in clinical and biomedical applications such as implants because of the high resistance of the composites. This review focuses on the effects of voltage, time and electrolytes on a calcium phosphate-base composite layer in case of pure titanium and other biomedical grade titanium alloys via the plasma electrolytic oxidation (PEO) method. Remarkably, these parameters changed the structure, morphology, pH, thickness and crystallinity of the obtained coating for various engineering and biomedical applications. Hence, the structured layer caused improvement of the biocompatibility, corrosion resistance and assignment of extra benefits for Osseo integration. The fabricated layer with a thickness range of 10 to 20 μm was evaluated for physical, chemical, mechanical and tribological characteristics via XRD, FESEM, EDS, EIS and corrosion analysis respectively, to determine the effects of the applied parameters and various electrolytes on morphology and phase transition. Moreover, it was observed that during PEO, the concentration of calcium, phosphor and titanium shifts upward, which leads to an enhanced bioactivity by altering the thickness. The results confirm that the crystallinity, thickness and contents of composite layer can be changed by applying thermal treatments. The corrosion behavior was investigated via the potentiodynamic polarization test in a body-simulated environment. Here, the optimum corrosion resistance was obtained for the coating process condition at 500 V for 15 min in Ringer solution. This review has been summarized, aiming at the further development of PEO by producing more adequate titanium-base implants along with desired mechanical and biomedical features., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The analysis of body fluids is of utmost importance in forensic casework since many biological fluids contain DNA. The ATR FT-IR spectroscopy is an emerging approach for the confirmatory, rapid, facile, non-destructive, and on-site identification and differentiation of body fluid stains. Notwithstanding the ATR FT-IR spectroscopy is showing a colossal promise towards the identification of body fluids, and further forensic enquiry related to substrate's interference is still in its infancy stage. Therefore, in the present proof-of-concept study, the ATR FT-IR spectroscopy has been utilized for the detection of vaginal fluid stains and to investigate the effect of different substrates on sample analysis. Simulated vaginal fluid samples were prepared on some selected substrates such as glass, plastic, floor tiles, polished wood, paper, and on various cloth substrates and analyzed without any prior sample preparation. Results suggested that vaginal fluid can be successfully detected on non-porous substrates, but it turned out to be a challenging task on porous substrates. However, on the basis of certain peaks, successful identification of vaginal fluid can be done directly on various case-related substrates. The best approach for the detection of vaginal fluid depends upon the nature of substrates and type of interference encountered. In addition, 10 non-vaginal fluid substances which look similar to vaginal fluid and which may lead to misclassification of vaginal fluid or can deliver false-positive results were also analyzed. The spectra of look-alike substances were classified using the chemometric tools such as PCA and PCA-LDA. The developed PCA model successfully classified all vaginal fluid samples from non-vaginal fluid substances with 100% accuracy, specificity, and sensitivity rate. In addition, the effects of other factors such as aging and mixing with other body fluids have also been studied and the results have been described.

The exploration into nanomaterial-based nonenzymatic biosensors with superb performance in terms of good sensitivity and anti-interference ability in disease marker monitoring has always attained undoubted priority in sensing systems. In this work, we report the design and synthesis of a highly active nanocatalyst, i.e., palladium and platinum nanoparticles (Pt&Pd-NPs) decorated ultrathin nanoporous gold (NPG) film, which is modified on a homemade graphene paper (GP) to develop a high-performance freestanding and flexible nanohybrid electrode. Owing to the structural characteristics the robust GP electrode substrate, and high electrochemically catalytic activities and durability of the permeable NPG support and ultrafine and high-density Pt&Pd-NPs on it, the resultant Pt&Pd-NPs–NPG/GP electrode exhibits excellent sensing performance of low detection limitation, high sensitivity and anti-interference capability, good reproducibility and long-term stability for the detection of small molecular biomarkers hydrogen peroxide (H2O2) and glucose (Glu), and has been applied to the monitoring of H2O2 in different types of live cells and Glu in body fluids such as urine and fingertip blood, which is of great significance for the clinical diagnosis and prognosis in point-of-care testing. [ABSTRACT FROM AUTHOR]

[Display omitted] • Triangular silver nanoprism (AgNPrs) was synthesized using novel hydrothermal protocol. • AgNPrs was used for the identification of dopamine in human biofluids using colorimetric and spectrophotometric assay. • A novel photochemical one-drop microarray was proposed for the low-cost sensing of dopamine. Trace-level detection of Dopamine (DA) in human body fluids is crucial for diagnosing neurological disorders, understanding mental health, and advancing drug discovery, but found challenging with rapid and low-cost detection technologies. Herein, an innovative paper-based photochemical method has been developed for the detection of DA using triangular silver nanoprism (AgNPrs), as an optical nanoprobe, in human blood plasma and urine samples. The redox reaction between DA and AgNPrs was evaluated through a color change at two pHs (5.35and 6.14), which can be analyzed by UV–Vis spectrophotometry and microfluidic paper-based colorimetric device (µ PCD) with low limit of quantification of 0.01 µM. This study presents pioneering work with the potential to inspire the development of simple and effective on-site semi quantifying recognition devices for other neurotransmitters, requiring only a lightweight piece of paper and a single drop of the necessary reagent. [ABSTRACT FROM AUTHOR]

Frequent oceanic oil spill incidents and the discharge of industrial oily wastewaters have caused serious threats to environments, food chains and human beings. Lignin wastes with many reactive groups exist as the byproducts from bioethanol and pulping processing industries, and they are either discarded as wastes or directly consumed as a fuel. To make full use of lignin wastes and simultaneously deal with oily wastewaters, porous lignin-based composites have been rationally designed and prepared. In this review, recent advances in the preparation of porous lignin-based composites are summarized in terms of aerogels, sponges, foams, papers, and membranes, respectively. Then, the mechanisms and the application of porous lignin-based adsorbents and filtration materials for oil/water separation are discussed. Finally, the challenges and perspectives of porous lignin-based composites are proposed in the field of oil/water separation. The utilization of abundant lignin wastes can replace fossil resources, and meanwhile porous lignin-based composites can be used to efficiently treat with oily wastewaters. The above utilization strategy opens an avenue to the rational design and preparation of lignin wastes with high-added value, and gives a possible solution to use lignin wastes in a sustainable and environmentally friendly way., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A well-developed-multiwall carbon nanotube (f-MWCNT)/biphasic calcium phosphate (BCP) composites were synthesized using ultrasonication method for orthopedic implantation applications. The formation of composites and its phase was confirmed by using X-ray diffraction. The presence of various functional groups was identified by using Fourier transform infra-red (FT-IR) spectroscopy. The presence of f-MWCNT was confirmed by Raman spectroscopy. High-resolution transmission electron microscopy (HR-TEM) analysis revealed that BCP units were bound by the surface of f-MWCNTs. The synthesized composites were coated on medical grade 316L stainless steel substrates using electro deposition technique. To determine its corrosion resistance characteristics, the developed substrates were exposed to a simulated bodily fluid (SBF) solution for 0, 4, and 7 days. These results strongly suggest that the coated composites can be utilized for bone tissue repair., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A significant bottleneck exists for mass-production of ion-selective electrodes despite recent developments in manufacturing technologies. Here, we present a fully-automated system for large-scale production of ISEs. Three materials, including polyvinyl chloride, polyethylene terephthalate and polyimide, were used as substrates for fabricating ion-selective electrodes (ISEs) using stencil printing, screen-printing and laser engraving, respectively. We compared sensitivities of the ISEs to determine the best material for the fabrication process of the ISEs. The electrode surfaces were modified with various carbon nanomaterials including multi-walled carbon nanotubes, graphene, carbon black, and their mixed suspensions as the intermediate layer to enhance sensitivities of the electrodes. An automated 3D-printed robot was used for the drop-cast procedure during ISE fabrication to eliminate manual steps. The sensor array was optimized, and the detection limits were 10 -5  M, 10 -5  M and 10 -4  M for detection of K + , Na + and Ca 2+ ions, respectively. The sensor array integrated with a portable wireless potentiometer was used to detect K + , Na + and Ca 2+ in real urine and simulated sweat samples and results obtained were in agreement with ICP-OES with good recoveries. The developed sensing platform offers low-cost detection of electrolytes for point-of-care applications., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Abstract Body fluid biomarkers have great potential for different clinical purposes, including diagnosis, prognosis, patient stratification and treatment effect monitoring. This is exemplified by current use of several excellent biomarkers, such as the Alzheimer’s disease cerebrospinal fluid (CSF) biomarkers, anti-neuromyelitis optica antibodies and blood neurofilament light. We still, however, have a strong need for additional biomarkers and several gaps in their development and implementation should be filled. Examples of such gaps are i) limited knowledge of the causes of neurological diseases, and thus hypotheses about the best biomarkers to detect subclinical stages of these diseases; ii) the limited success translating discoveries obtained by e.g. initial mass spectrometry proteomic low-throughput studies into immunoassays for widespread clinical implementation; iii) lack of interaction among all stakeholders to optimise and adapt study designs throughout the biomarker development process to medical needs, which may change during the long period needed for biomarker development. The Society for CSF Analysis and Clinical Neurochemistry (established in 2015) has been founded as a concerted follow-up of large standardisation projects, including BIOMARKAPD and SOPHIA, and the BioMS-consortium. The main aims of the CSF society are to exchange high level international scientific experience, to facilitate the incorporation of CSF diagnostics into clinical practice and to give advice on inclusion of CSF analysis into clinical guidelines. The society has a broad scope, as its vision is that the gaps in development and implementation of biomarkers are shared among almost all neurological diseases and thus they can benefit from the activities of the society.

The homeostatic control of Sodium (Na+) ion in the human body assumes paramount relevance owing to its physiological importance. Any deviation from the normal level causes serious health problems like hypernatremia, hyponatremia, stroke, kidney problems etc. Therefore, quantification of Na+ levels in body fluids has significant diagnostic and prognostic importance. However, interfering ions like Potassium ion (K+) is the major hurdle in sodium detection. In this work, we synthesized the clusters of 3–9 nm-sized highly stable and pure Copper nanoparticles surface functionalised with curcumin, through chemical reduction method. Each cluster of particles is encapsulated in a curcumin layer which is clearly visible in TEM images. The results show that these curcumin functionalized Cu NPs (CuC) are highly selective to the colorimetric detection of Na+. The ions like K+, Mg2+ and Zn2+ did not interfere with the Na+ in this sensing technique. Low-cost paper-based sensor strips are fabricated and calibrated for the sensing of sodium in the physiological range and shade cards were developed as a calorimetric guide for estimation of Na+ which makes them ideal point of care diagnostic platform. We demonstrate that the proposed CuC paper strip can be used for detecting Na+ concentration within the whole physiological range in both blood serum and urine. [ABSTRACT FROM AUTHOR]

Background: The use of filter paper as a simple, inexpensive tool for storage and transportation of blood, ‘Dried Blood Spots’ or Guthrie cards, for diagnostic assays is well-established. In contrast, there are a paucity of diagnostic evaluations of dried cerebrospinal fluid (CSF) spots. These have potential applications in low-resource settings, such as Laos, where laboratory facilities for central nervous system (CNS) diagnostics are only available in Vientiane. In Laos, a major cause of CNS infection is Japanese encephalitis virus (JEV). We aimed to develop a dried CSF spot protocol and to evaluate its diagnostic performance using the World Health Organisation recommended anti-JEV IgM antibody capture enzyme-linked immunosorbent assay (JEV MAC-ELISA). Methodology and Principal Findings: Sample volumes, spotting techniques and filter paper type were evaluated using a CSF-substitute of anti-JEV IgM positive serum diluted in Phosphate Buffer Solution (PBS) to end-limits of detection by JEV MAC-ELISA. A conventional protocol, involving eluting one paper punch in 200μl PBS, did not detect the end-dilution, nor did multiple punches utilising diverse spotting techniques. However, pre-cut filter paper enabled saturation with five times the volume of CSF-substitute, sufficiently improving sensitivity to detect the end-dilution. The diagnostic accuracy of this optimised protocol was compared with routine, neat CSF in a pilot, retrospective study of JEV MAC-ELISA on consecutive CSF samples, collected 2009–15, from three Lao hospitals. In comparison to neat CSF, 132 CSF samples stored as dried CSF spots for one month at 25–30°C showed 81.6% (65.7–92.3 95%CI) positive agreement, 96.8% (91.0–99.3 95%CI) negative agreement, with a kappa coefficient of 0.81 (0.70–0.92 95%CI). Conclusions/Significance: The novel design of pre-cut filter paper saturated with CSF could provide a useful tool for JEV diagnostics in settings with limited laboratory access. It has the potential to improve national JEV surveillance and inform vaccination policies. The saturation of filter paper has potential use in the wider context of pathogen detection, including dried spots for detecting other analytes in CSF, and other body fluids. [ABSTRACT FROM AUTHOR]

The Curfews period of December 2015 to February 2016 in south-eastern Turkey was a time of collective violence in multiple forms, among which the Cizre Basement Massacres are the most known. The period of the general elections in June 2015, until the later and most recent attempt to carry out a military coup in July 2016, covers multiple curfews and military operations in the south-eastern regions of Turkey, and some social media material shared from the region caught international attention and provoked artistic as well as political responses. Zehra Doğan, a Kurdish journalist and artist, modified and re-shared some footage that was first posted in social media accounts of the special forces in Nusaybin and as a result was sentenced to three years in prison. She continued to produce artworks in prison, even though it was 'prohibited' for her to draw. She started to use her own body fluids, hair, newspaper, toilet paper, and any other available daily material in the prison to produce works of art. She also produced a series of work with her menstrual blood. For this article, I conducted two thematic one-to-one discussions with Zehra Doğan and studied her series of works produced with menstrual blood within the scope of mimesis and embodiment theories. Further sections of the article discuss: the function of these series of works for her acting out and working through processes of the collective political violence of which she became both a witness and a victim; methods of artistic transmission of political violence both for the artist and for the audience; and the possible ritualistic functions of using blood while producing work of arts in relation to collective political killings. [ABSTRACT FROM AUTHOR]

Competing Interests: The authors have no funding or conflicts of interest to disclose.

A novel SnO 2 quantum dots (SnO 2 QDs)-functionalized Ti 3 C 2 MXene nanocomposite was prepared via in situ synthesis method, resulting in well-regulated the nucleation and growth of SnO 2 QDs to evenly distribute onto MXene nanosheets. Ultra-small size SnO 2 QDs decorated on the surface of Ti 3 C 2 MXene nanosheets can effectively prevent the restacking of MXene and remarkably increase the electroactive surface area of the electrode, which can further increase electrocatalytic activity toward dopamine. Then, an ultrasensitive electroanalytical method based on SnO 2 QDs-functionalized Ti 3 C 2 MXene nanocomposite for dopamine monitoring was developed, and the effects of experimental condition were investigated systematically. Under optimized conditions, the prepared sensor presented a linear dependence for dopamine in the concentration range from 0.004 to 8.0 µM with the detection limit of 2.0 nM (S/N = 3). Moreover, it selectively perceived dopamine in presence of physiological interferents in urine and serum samples with excellent linearities (correlation coefficients higher than 0.9920). The relative recoveries were in the range 97.67-105.3% and 103.0-106.8%, while the limits of quantitation were 10.12 nM and 9.62 nM in urine and serum sample, respectively, demonstrating the method suitability for dopamine sensing and being envisioned as a promising candidate for neurotransmitter monitoring in biological diagnosis., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Contemporary criminal investigations are based on the statements made by the victim and the eyewitnesses. They also rely on the physical evidences found in the crime scene. These evidences, and more particularly biological ones, have a great judicial value in the courtroom. They are usually used to revoke the suspect's allegations and confirm or refute the statements made by the victim and the witnesses. Stains of body fluids are biological evidences highly sought by forensic investigators. In many criminal cases, the success of the investigation relies on the correct identification and classification of these stains. Therefore, the adoption of reliable and accurate forensic analytical methods seems to be of vital importance to attain this objective. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) is a modern and universal analytical technique capable of fingerprint recognition of the analyte using minimal amount of the test sample. The current systematic review aims to through light on the fundamentals of this technique and to illustrate its wide range of applications in forensic investigations. ATR-FTIR is a nondestructive technique which has demonstrated an exceptional efficiency in detecting, identifying and discriminating between stains of various types of body fluids usually encountered in crime scenes. The ATR-FTIR spectral data generated from bloodstains can be used to deduce a wealth of information related to the donor species, age, gender, and race. These data can also be exploited to discriminate between stains of different types of bloods including menstrual and peripheral bloods. In addition, ATR-FTIR has a great utility in the postmortem investigations. More particularly, in estimating the postmortem interval and diagnosing death caused by extreme weather conditions. It is also useful in diagnosing some ambiguous death causes such as fatal anaphylactic shock and diabetic ketoacidosis., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cancer is one of the leading causes of death worldwide producing an estimated cost of $161.2 billion in the US in 2017 only. Early detection of cancer can not only reduce cancer mortality rates but also dramatically reduce healthcare costs given that the 17 million new cancer cases in 2018 are estimated to grow 27.5 million new cases by 2040. Analytical devices based upon paper substrates can provide effective, rapid, and extremely low‐cost alternatives for early cancer detection compared to existing testing methods. However, low concentrations of biomarkers in body fluids as well as the possible association of any given biomarker with multiple diseases remains one of the greatest challenges to widespread adoption of these paper‐based devices. Fortunately, recent advances have opened the possibility of detecting multiple biomarkers within the same device, which can be predictive of a patient's condition with unprecedented cost‐effectiveness. Accordingly, this review highlights the recent advancements in paper‐based analytical devices with a multiplexing focus. The primary areas of interest include lateral flow assay and microfluidic paper‐based assay formats, signal amplification approaches to enhance the sensitivity for a specific cancer type, along with current challenges and future outlook for the detection of multiple cancer biomarkers. [ABSTRACT FROM AUTHOR]

Paper-based biofuel cells have the potential to serve as health monitoring devices for urinary glucose and volume by incorporating the fabricated electrodes into nursing diapers. In this study, we evaluate the output power and stability of a biofuel cell by combining 3D impedance measurement and admittance analysis. The 3D impedance-based analysis method can simultaneously detect changes in the overall electrode structure of the enzyme electrode and in the quantity of active enzymes and mediators on the electrode surface. The biofuel cell electrodes were fabricated via screen-printing, using polydimethylglycidyl (poly-GMA)- modified MgO-templated carbon (GMgOC) or poly-GMA-unmodified MgO-templated carbon (NMgOC). The enzymes used were flavin adenine dinucleotide-dependent glucose dehydrogenase and bilirubin oxidase, with Azure-A serving as the mediator. Admittance analysis was utilized to measure the enzyme activity and estimate the charge transfer resistance by extrapolating the spectra obtained through admittance analysis. The charge transfer resistances of both electrodes exhibited a greater stability for the GMgOC electrode. This can be attributed to the covalent immobilization of the enzyme and mediator on the electrode facilitated by GMgOC, effectively suppressing enzyme and mediator elution. The electric double-layer capacitance values of both electrodes indicated the overall stability of the electrode structure during the measurement. [ABSTRACT FROM AUTHOR]

Saliva is a common body fluid with significant forensic value used to investigate criminal cases such as murder and assault. In the past, saliva identification often relied on the α-amylase test; however, this method has low specificity and is prone to false positives. Accordingly, forensic researchers have been working to find new specific molecular markers to refine the current saliva identification approach. At present, research on immunological methods, mRNA, microRNA, circRNA, and DNA methylation is still in the exploratory stage, and the application of these markers still has various limitations. It has been established that salivary microorganisms exhibit good specificity and stability. In this study, 16S rDNA sequencing technology was used to sequence the V3-V4 hypervariable regions in saliva samples from five regions to reveal the role of regional location on the heterogeneity in microbial profile information in saliva. Although the relative abundance of salivary flora was affected to a certain extent by geographical factors, the salivary flora of each sample was still dominated by Streptococcus, Neisseria, and Rothia. In addition, the microbial community in the saliva samples in this study was significantly different from that in the vaginal secretions, semen, and skin samples reported in our previous studies. Accordingly, saliva can be distinguished from the other three body fluids and tissues. Moreover, we established a prediction model based on the random forest algorithm that could distinguish saliva between different regions at the genus level even though the model has a certain probability of misjudgment which needs more in-depth research. Overall, the microbial community information in saliva stains might have prospects for potential application in body fluid identification and biogeographic inference., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The transition period is very stressful for primiparous cows due to their first calving experience and will be more challenging if it occurs under heat stress conditions. Heat stress reduces the feed intake of dairy cows. Therefore, it reduces the consumption of minerals and vitamins. Oral administration of boluses through the provision of mineral-vitamin compounds can reduce metabolic abnormalities after calving. The present study aimed to evaluate effect of sustained-release bolus on body condition score (BCS) change, serum metabolites, uterine health, and reproductive status in primiparous cows. Heifers were selected at the beginning of the close-up period (n = 60, BCS = 3.35 ± 0.12). There were 2 experimental treatments at the time of calving: (1) heifers without bolus oral administration (H - Bo, n = 30); (2) heifers with bolus oral administration (H + Bo, n = 30). The results showed that although the rate of BCS loss was lower in the group receiving bolus, the effect of bolus was not significant. The effect of bolus on blood level of glucose, non-esterified fatty acids (NEFA), and beta-hydroxybutyrate (BHBA) was not significant; however, the highest concentration of albumin (P = 0.05) was observed in the H + Bo group on day 42 after calving. The concentration of aspartate transaminase (AST) tended to increase (P = 0.06) on day 14 after calving and entire the study. Total antioxidant capacity (TAC) was affected (P < 0.05) by bolus throughout the period of study, and the highest (P < 0.05) concentration of glutathione peroxidase (GPX) and superoxide dismutase (SOD) was observed in H + Bo group on day 42 after calving. The H + Bo group had the lowest (P < 0.05) vaginal discharge score (VDS). In general, oral administration of the sustained-release bolus in heifers significantly affected the antioxidant factors and uterine health, as well as had positive effects on liver function, body condition, and reproduction status., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Practical screening tools and ultrasensitive technologies can play pivotal roles in precision cancer profiling for early diagnosis at asymptomatic stages, as well as for monitoring prognosis, risk stratification, and disease recurrence. While a number of sensors and diagnostic tools continue to be developed for ultrasensitive detection and off-site analysis, there has been an increasing interest in point-of-care devices, particularly those that are mechanically flexible and potentially wearable by the patient. In this chapter, we present a critical insight into the integrated engineering approaches involved in such flexible systems. We consider various aspects in the design of flexible devices, the biomarkers of interest, and the different transduction mechanisms by which mechanically flexible devices can be used in the area of cancer monitoring. We then discuss the different types of flexible biosensing platforms that have been developed to date, including wearables on skin and on clothing, and exhaled breath and implantable sensors. Finally, we discuss the design challenges and future outlook in the development of flexible platforms that can provide comprehensive cancer biomarker panels for patients and clinicians., (© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

Over the past decades, paper-based lateral flow immunoassays (LFIAs) have been extensively developed for rapid, facile, and low-cost detection of a wide array of target analytes in a point-of-care manner. Conventional home pregnancy tests are the most significant example of LFAs, which detect elevated concentrations of human chorionic gonadotrophin (hCG) in body fluids to identify early pregnancy. In this work, we have upgraded these platforms to a higher version by developing a customized microfluidic paper-based analytical device (μPAD), as the new generation of paper-based point-of-care platforms, for colorimetric immunosensing. This will offer a cost-efficient and environmentally friendly alternative platform for paper-based immunosensing, eliminating the need for nitrocellulose (NC) membrane as the substrate material. The performance of the developed platform is demonstrated by detection of hCG (as a model case) in urine samples and subsequently indicating positive or negative pregnancy. A dual-functional silane-based composite was used to treat filter paper in order to enhance the colorimetric signal intensity in the detection zones of μPADs. In addition, microfluidic pathways were designed in a manner to provide the desired regulated fluid flow, generating sufficient incubation time (delays) at the designated detection zones, and consequently enhancing the obtained signal intensity. The presented approaches allow to overcome the existing limitations of μPADs in immunosensing and will broaden their applicability to a wider range of assays. Although, the application of the developed hCG μPAD assay is mainly in qualitative (i.e., positive or negative) detection of pregnancy, the semi-quantitative measurement of hCG was also investigated, indicating the viability of this assay for sensitive detection of the target hCG analyte within the related physiological range (i.e., 10–500 ng/mL) with a LOD value down to 10 ng/mL. [ABSTRACT FROM AUTHOR]

6-Thioguanine is an immunosuppressive drug, an analogue of guanine, applied to treat acute leukemia and inflammatory bowel disease. Excessive use of 6-thioguanine during clinical treatment may cause side effects. Moreover, providing a dose too low will be ineffective. Therefore, there is a critical need for a rapid, selective and routine approach to quantifying 6-thioguanine in body fluids to support a clinical application. A fully validated HPLC method has been developed to determine 6-thioguanine in whole blood samples using 5-bromouracil as an internal standard. 6-Thioguanine nucleotides were released from erythrocytes by perchloric acid, and then hydrolysed at 100 °C to the parent thiopurine, 6-thioguanine. The following validation parameters of the method were determined: specificity/selectivity, linearity range (479-17,118 ng/mL, R > 0.992), limits of detection (150 ng/mL) and quantification (479 ng/mL), accuracy (- 5.6 < Bias < 14.7), repeatability (CV 1.30-3.24%), intermediate precision (CV 4.19-5.78%), extraction recovery (79.1-103.6%) and carryover. Furthermore, the stability of the drug in whole blood samples under various storage conditions was investigated. The suggested method is suitable for determining 6-thioguanine in whole blood erythrocyte samples for drug level monitoring, thus correct dosing., (© 2023. Springer Nature Limited.)

Human body fluids contain biomarkers which are used extensively for prognostication, diagnosis, monitoring, and evaluation of different treatments for a variety of diseases and disorders. The application of biosensors based on cell-free protein synthesis (CFPS) offers numerous advantages including on-demand and at-home use for fast, accurate detection of a variety of biomarkers in human fluids at an affordable price. However, current CFPS-based biosensors use commercial RNase inhibitors to inhibit different RNases present in human fluids and this reagent is approximately 90% of the expense of these biosensors. Here the flexible nature of Escherichia coli-lysate-based CFPS was used for the first time to produce murine RNase Inhibitor (m-RI) and to optimize its soluble and active production by tuning reaction temperature, reaction time, reduced potential, and addition of GroEL/ES folding chaperons. Furthermore, RNase inhibition activity of m-RI with the highest activity and stability was determined against increasing amounts of three human fluids of serum, saliva, and urine (0%-100% v/v) in lyophilized CFPS reactions. To further demonstrate the utility of the CFPS-produced m-RI, a lyophilized saliva-based glutamine biosensor was demonstrated to effectively work with saliva samples. Overall, the use of CFPS-produced m-RI reduces the total reagent costs of CFPS-based biosensors used in human body fluids approximately 90%., (© 2021 Wiley Periodicals LLC.)

Simultaneous detection of multiple tumor biomarkers in body fluids could facilitate early diagnosis of lung cancer, so as to provide scientific reference for clinical treatment. This paper depicted a multi-parameter paper-based electrochemical aptasensor for simultaneous detection of carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in a clinical sample with high sensitivity and specificity. The paper-based device was fabricated through wax printing and screen-printing, which enabled functions of sample filtration and sample auto injection. Amino functional graphene (NG)-Thionin (THI)- gold nanoparticles (AuNPs) and Prussian blue (PB)- poly (3,4- ethylenedioxythiophene) (PEDOT)- AuNPs nanocomposites were synthesized respectively. They were used to modify the working electrodes not only for promoting the electron transfer rate, but also for immobilization of the CEA and NSE aptamers. A label-free electrochemical method was adopted, enabling a rapid simple point-of-care testing. Experimental results showed that the proposed multi-parameter aptasensor exhibited good linearity in ranges of 0.01–500 ng mL−1 for CEA (R2 = 0.989) and 0.05–500 ng mL−1 for NSE (R2 = 0.944), respectively. The limit of detection (LOD) was 2 pg mL−1 for CEA and 10 pg mL−1 for NSE. In addition, the device was evaluated using clinical serum samples and received a good correlation with large electrochemical luminescence (ECL) equipment, which would offer a new platform for early cancer diagnostics, especially in those resource-limit areas. • The aptasensor could enable simultaneous detection of two tumor markers in a single sample with high sensitivity. • The aptasensor had a fast response time as a label-free electrochemical detection method was adopted. • The aptasensor was fabricated by wax printing and screen-printing, which could be mass-produced easily with low cost. [ABSTRACT FROM AUTHOR]

Background: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTATM Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo’s national surveillance for routine diagnosis were selected for this study. Methods: The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory’s standard DNA extraction, and processing the same samples after being dried and stored on FTATM card, and DNA extracted following the manufacturer’s instructions. Results: The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTATM cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory’s standard methodology, results showed high concordance, with Kappa index ranges of 0.9877–1.00 for CSF, and 0.8004–1.00 for serum samples. Conclusion: The use of FTATM cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost. [ABSTRACT FROM AUTHOR]

Exosomes emerged as valuable sources of disease biomarkers and new therapeutic tools. However, extracellular vesicles isolation with exosome-like characteristics from certain biofluids is still challenging which can limit their potential use in clinical settings. While ultracentrifugation-based procedures are the gold standard for exosome isolation from cell cultures, no unique and standardized method for exosome isolation from distinct body fluids exists. The complexity, specific composition, and physical properties of each biofluid constitute a technical barrier to obtain reproducible and pure exosome preparations, demanding a detailed characterization of both exosome isolation and characterization methods. Moreover, some isolation procedures can affect downstream proteomic or RNA profiling analysis. This review compiles and discussed a set of comparative studies addressing distinct exosome isolation methods from human biofluids, including cerebrospinal fluid, plasma, serum, saliva, and urine, also focusing on body fluid specific challenges, physical properties, and other potential variation sources. This summarized information will facilitate the choice of exosome isolation methods, based on the type of biological samples available, and hopefully encourage the use of exosomes in translational and clinical research., (© 2022. Springer-Verlag GmbH Germany, part of Springer Nature.)

Extracellular vesicles (EV) are small membrane-coated structures secreted by all cells of the body and can be detected in all bodily fluids, including urine. EV contents (e.g. proteins and distinct RNA classes) reflect the physiological state of their cells of origin, offering a new source of biomarkers. Accordingly, urinary Extracellular Vesicles (uEVs) are emerging as a source for early biomarkers of kidney damage and beyond, holding the potential to replace the conventional invasive techniques including kidney biopsy. However, the lack of standardization and sample collection and isolation methods, and the influence of factors such as inter- and intra-individual variability create difficulties in interpreting current results. Here we review recent results and reported uses of especially urinary EVs and also pinpoint approaches to be considered when designing experiments.

Extracellular vesicles (EVs) are considered as cargo and mediate intercellular communication. As natural biological nanoparticles, EVs can be secreted by almost all kinds of cells and exist in biofluids such as milk, urine, blood, etc. In the past decades, several methods have been utilized to isolate EVs from cell culture medium, biofluids, and tissues. Here in this chapter, we summarized conventional and novel methods and fundamental procedures of EVs extraction and purification from different biofluids (plasma, urine, milk, and saliva) and tissues (brain, intestinal tissue, muscles, and heart). The present section also discusses how to choose appropriate methods to extract EVs from tissues based on downstream analysis. This chapter will expand the horizons of EVs isolation and purification from different mediums., (© 2023. The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd.)

Coronary artery disease (CAD) is the leading cause of death and disability worldwide. Despite recent progress in the diagnosis and treatment of CAD, evidence gaps remain, including pathogenesis, the most efficient diagnostic strategy, prognosis of individual patients, monitoring of therapy, and novel therapeutic strategies. These gaps could all be filled by developing novel, minimally invasive, blood-based biomarkers. Potentially, extracellular vesicles (EVs) could fill such gaps. EVs are lipid membrane particles released from cells into blood and other body fluids. Because the concentration, composition, and functions of EVs change during disease, and because all cell types involved in the development and progression of CAD release EVs, currently available guidelines potentially enable reliable and reproducible measurements of EVs in clinical trials, offering a wide range of opportunities. In this chapter, we provide an overview of the associations reported between EVs and CAD, including (1) the role of EVs in CAD pathogenesis, (2) EVs as biomarkers to diagnose CAD, predict prognosis, and monitor therapy in individual patients, and (3) EVs as new therapeutic targets and/or drug delivery vehicles. In addition, we summarize the challenges encountered in EV isolation and detection, and the lack of standardization, which has hampered real clinical applications of EVs. Since most conclusions are based on animal models and single-center studies, the knowledge and insights into the roles and opportunities of EVs as biomarkers in CAD are still changing, and therefore, the content of this chapter should be seen as a snapshot in time rather than a final and complete compendium of knowledge on EVs in CAD., (© 2023. The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd.)

Synthetic cannabinoids (SCs) remain one of the largest groups of new psychoactive substances. Recently, new synthetic cannabinoids 5F-MDMB-PICA and 4F-MDMB-BINACA are increasing in popularity. A 33-year-old man lost consciousness after smoking an unknown substance. A glass pipe and two lumps of substance that turned out to contain 5F-MDMB-PICA and 4F-MDMB-BINACA were found at the scene. Blood, urine and cerebrospinal fluid were collected during the examination of the body. The synthetic cannabinoids were isolated from autopsy materials by precipitation with acetonitrile and extraction with ethyl acetate. The screening and quantitative analyses were performed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The liquid chromatography-quadrupole/time of flight mass spectrometry (LC-Q/TOF) technique was used for metabolite identification. 5F-MDMB-PICA was detected and quantified in all analysed materials, whereas 4F-MDMB-BINACA was found only in cerebrospinal fluid. The determined concentrations of 5F-MDMB-PICA were 0.9 (blood), 0.1 (urine) and 3.2 ng/mL (cerebrospinal fluid). The concentration of 4F-MDMB-BINACA in cerebrospinal fluid was 0.1 ng/mL. The main metabolites of both compounds (hydrolysis and oxidative defluorination) were found in all analysed body fluids. Cerebrospinal fluid may be important alternative material in autopsy cases. Rapid elimination of 5F-MDMB-PICA and 4F-MDMB-BINACA compounds also means that the metabolite analysis can be crucial for the investigation. Laboratories must be made aware of their presence and incorporate these SCs and their metabolites into workflows for detection and confirmation. Ester hydrolysis and oxidative defluorination products can be found in blood, urine and cerebrospinal fluid making them useful biomarkers of intake., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Metastasis is a dynamic succession of events involving the dissemination of tumour cells to distant sites within the body, ultimately reducing the survival of patients with cancer. To colonize distant organs and, therefore, systemically disseminate within the organism, cancer cells and associated factors exploit several bodily fluid systems, which provide a natural transportation route. Indeed, the flow mechanics of the blood and lymphatic circulatory systems can be co-opted to improve the efficiency of cancer cell transit from the primary tumour, extravasation and metastatic seeding. Flow rates, vessel size and shear stress can all influence the survival of cancer cells in the circulation and control organotropic seeding patterns. Thus, in addition to using these fluids as a means to travel throughout the body, cancer cells exploit the underlying physical forces within these fluids to successfully seed distant metastases. In this Review, we describe how circulating tumour cells and tumour-associated factors leverage bodily fluids, their underlying forces and imposed stresses during metastasis. As the contribution of bodily fluids and their mechanics raises interesting questions about the biology of the metastatic cascade, an improved understanding of this process might provide a new avenue for targeting cancer cells in transit.

The illegal use of clenbuterol seriously endangers food safety and human health. Accurate monitoring of the illegal use of clenbuterol in livestock can efficiently prevent the clenbuterol residue pork products from entering the consumer market. Thus, in this study, a simple, rapid, and sensitive method for the determination of clenbuterol in swine urine was developed using electromembrane extraction combined with liquid chromatography-tandem mass spectrometry. It should be noted that the electromembrane extraction method presented many advantages of simple operation, fast mass transfer rate, good sample clean-up capability, and less organic solvent consumption. The effect of important factors on the extraction efficiency of clenbuterol was investigated. Under the optimal conditions, good linearity was achieved for clenbuterol over the range of 1-1000 ng/ml (linear correlation [R 2 ] = 0.9996). The recoveries of clenbuterol in swine urine at three spiked levels ranged from 83.7% to 110.0% with relative standard deviation values lower than 9.7% (n = 4). The limits of detection and quantification for clenbuterol were 0.07 and 0.25 ng/ml, respectively. These results suggested that the proposed method has great potential for the extraction and determination of trace analyte in a complex sample matrix for monitoring illegal use in livestock., (© 2022 Wiley-VCH GmbH.)

The characteristics of ATR FT-IR spectroscopy are extremely attractive and escalating popularity in the field of body fluid analysis owing to its non-destructive, rapid, and reliable nature. Herein, the present study establishes that how ATR FT-IR spectroscopy could be utilized as a non-destructive, non-invasive, and confirmatory technique for sex discrimination from dry urine traces. Traces of body fluids are of paramount importance to criminal investigations as a major source of individualization by DNA profiling. However, the significance of DNA profiling from urine traces is highly diminished due to the small amount of DNA in urine traces. For that reason, the sex discrimination between the male and female donors is sorely desirable. In this study, ATR FT-IR spectroscopy in combination with partial least squares-discriminant analysis (PLS-DA) model unequivocally demonstrated the successful sex discrimination of an individual from dry traces of urine with 95.3% accuracy. PCA-Linear Discriminant Analysis (LDA) approach provided 85.2% of accuracy; however, PCA could not provide the sufficient findings for the discrimination of male and female urine spectra. The validation study was conducted and obtained 0% rates of false-positive and negative assignments. Additionally, this study also attended to assess the influence of substrates on the analysis of urine traces and results have been discussed., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Identification of the source of mosquito blood meals is an important component for disease control and surveillance. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has emerged as an effective tool for mosquito blood meal identification, using the abdomens of freshly engorged mosquitoes. In the field, mosquito abdomens are crushed on Whatman filter papers to determine the host feeding patterns by identifying the origin of their blood meals. The aim of this study was to test whether crushing engorged mosquito abdomens on Whatman filter papers was compatible with MALDI-TOF MS for mosquito blood meal identification. Both laboratory reared and field collected mosquitoes were tested. Material and methods: Sixty Anopheles gambiae Giles were experimentally engorged on the blood of six distinct vertebrate hosts (human, sheep, rabbit, dog, chicken and rat). The engorged mosquito abdomens were crushed on Whatman filter papers for MALDI-TOF MS analysis. 150 Whatman filter papers, with mosquitoes engorged on cow and goat blood, were preserved. A total of 77 engorged mosquito abdomens collected in the Comoros Islands and crushed on Whatman filter papers were tested with MALDI-TOF MS. Results: The MS profiles generated from mosquito engorged abdomens crushed on Whatman filter papers exhibited high reproducibility according to the original host blood. The blood meal host was correctly identified from mosquito abdomens crushed on Whatman filter papers by MALDI-TOF MS. The MS spectra obtained after storage were stable regardless of the room temperature and whether or not they were frozen. The MS profiles were reproducible for up to three months. For the Comoros samples, 70/77 quality MS spectra were obtained and matched with human blood spectra. This was confirmed by molecular tools. Conclusion: The results demonstrated that MALDI-TOF MS could identify mosquito blood meals from Whatman filter papers collected in the field during entomological surveys. The application of MALDI-TOF MS has proved to be rapid and successful, making it a new and efficient tool for mosquito-borne disease surveillance. [ABSTRACT FROM AUTHOR]

Type 1 diabetes (T1D) is increasing in incidence and predictable with measurement of serum islet autoantibodies (iAb) years prior to clinical disease onset. Identifying iAb positive individuals reduces diabetic ketoacidosis and identifies individuals for T1D prevention trials. However, large scale screening for iAb remains challenging as assays have varying sensitivities and specificities, insulin autoantibodies remain difficult to measure and venipuncture is generally required to obtain serum. We developed an approach to reliably measure all four major iAb, including insulin autoantibodies, from dried blood spots (DBS) on filter-paper. By spiking iAb positive serum into iAb negative whole blood in a dose titration, we optimized the conditions for autoantibody elution from filter paper as measured by fluid phase radioimmunoassays. After assessing stability of measuring iAb from DBS over time, we then screened iAb from DBS and the corresponding serum in new-onset T1D (n = 52), and controls (n = 72) which included first-degree relatives of T1D patients. iAb measured from eluted DBS in new-onset T1D strongly correlated with serum measurements (R2 = 0.96 for mIAA, GADA = 0.94, IA-2A = 0.85, ZnT8A = 0.82, p<0.01 for each autoantibody). There were no false positives in control subjects, and 5/6 with previously unknown iAb positivity in sera were detected using DBS. With further validation, measuring iAb from DBS can be a reliable method to screen for T1D risk. [ABSTRACT FROM AUTHOR]

India has the highest burden of Tuberculosis (TB) and multidrug-resistant TB (MDR-TB) worldwide. Innovative technology is the need of the hour to identify these cases that remain either undiagnosed or inadequately diagnosed due to the unavailability of appropriate tools at primary healthcare settings. We developed and evaluated 3 kits, namely ‘TB Detect’ (containing BioFM-Filter device), ‘TB Concentration and Transport’ (containing Trans-Filter device) and ‘TB DNA Extraction’ kits. These kits enable bio-safe equipment-free concentration of sputum on filters and improved fluorescence microscopy at primary healthcare centres, ambient temperature transport of dried inactivated sputum filters to central laboratories and molecular detection of drug resistance by PCR and DNA sequencing (Mol-DST). In a 2-site evaluation (n = 1190 sputum specimens) on presumptive TB patients, BioFM-Filter smear exhibited a significant increase in positivity of 7% and 4% over ZN smear and LED-FM smear (p<0.05), respectively and an increment in smear grade status (1+ or 2+ to 3+) of 16% over ZN smear and 20% over LED-FM smear. The sensitivity of Mol-DST in presumptive MDR-TB and XDR-TB cases (n = 148) was 90% for Rifampicin (95% confidence interval [CI], 78–96%), 84% for Isoniazid (95% CI, 72–92%), 83% for Fluoroquinolones (95% CI, 66–93%) and 75% for Aminoglycosides (95% CI, 35–97%), using phenotypic DST as the reference standard. Test specificity was 88–93% and concordance was ~89–92% (κ value 0.8–0.9). The patient-friendly kits described here address several of the existing challenges and are designed to provide ‘Universal Access’ to rapid TB diagnosis, including drug-resistant disease. Their utility was demonstrated by application to sputum at 2 sites in India. Our findings pave the way for larger studies in different point-of-care settings, including high-density urban areas and remote geographical locations. [ABSTRACT FROM AUTHOR]

An immunoassay was previously developed as a technique to improve methods for detection and analysis of fly artifacts found at crime scenes. The dot blot assay utilized a polyclonal antiserum (anti-md3) based on a unique digestive cathepsin D found in cyclorrhaphous Diptera. In this study, artifacts produced by adults of Calliphora vicina, Cynomya cadaverina, Sarcophaga bullata, and Protophormia terraenovae were examined using the immunoassay to determine if insect-derived stains could be distinguished from a range of human body fluid stains. A lift technique was developed which permitted transfer of fly artifacts from test materials to filter paper for dot blot analyses. All species readily deposited artifacts on all test household materials regardless of diet consumed. Despite differences in texture and porosity of the household materials, artifacts of all species transferred to the filter paper. With all fly species, anti-md3 serum bound to artifacts produced after feeding on semen, blood, feces, urine, and saliva. By contrast, anti-md3 serum did not react with any of the human fluids tested, nor with any of the lifts from household materials not exposed to flies. There was no evidence of false positives with any of the fly species tested, regardless of diet consumed. There was also no indication of false negatives with any of the dot blot assays. These observations suggest that immunoassays using anti-md3 serum performed on a simple lift of suspected fly artifacts can be used effectively as a confirmatory assay to distinguish fly regurgitate and fecal stains from human body fluids. [ABSTRACT FROM AUTHOR]