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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.

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.

Paper chromatographic procedures may be used to detect cephalothin and its metabolite desacetylcephalothin in urine, plasma, synovial fluid, and cerebrospinal fluid. Protein-bound antibiotics are released from plasma, cerebrospinal fluid, and synovial fluid by dilution with an equal volume of dimethylformamide. Data are presented on the sample preparation, paper chromatographic system, and other specific techniques.

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]

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]

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]

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.)

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.)

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]

In addition to cobalt, metallic nickel, cadmium and other metals (e.g. zinc and copper) dissolve when incubated with horse serum at 37°. The dissolving property of copper in serum resembles that of cobalt, its solubility being increased greatly in the presence of oxygen, whereas the solubilities of cadmium, zinc and most preparations of nickel are the same aerobically and anaerobically. In all of these "metal-sera" the cations are bound, although in different proportions, both by proteins and by small diffusible molecules.Although Co(2+) ions and cobalt-serum cause limited catalytic oxidation of fresh serum, most of the oxygen uptake by suspensions of metallic cobalt in serum, or by more simple model systems, is due to absorption of oxygen by the metal powder; the consequences of this are discussed.Metallic cobalt, cadmium and nickel dissolve readily when incubated with sterile homogenates of rat muscle (and other tissues), the dissolved cations being bound predominantly by small, diffusible molecules, rather than by the protein components. Binding by small molecules, in preference to proteins, also occurs when the metals dissolve in vivo. In both the in vivo and in vitro systems, the metallic ions are not bound by a specific cation carrier, but are distributed amongst a number of components. These components have greater affinities for the dissolved metals than serum proteins and seem likely to be the normal cation carriers in vivo. As in serum, solubility in muscle homogenates is not a specific property of the carcinogenic metals as other, non-carcinogenic metals also dissolve. The specificity of the former metals, therefore, is attributed to the subsequent effects of the dissolved cations after intracellular incorporation.

Methods using dialysis or ultrafiltration are described for the collection of extracellular fluid in rumen contents for analysis of amino acids. Marked differences in the concentration of aspartic acid, glutamic acid, and alanine were found in samples of either diffusate or ultrafiltrate and in clarified acidified rumen liquor. Concentrations are given for aspartic acid, glutamic acid, alanine, glycine, gamma-aminobutyric acid, valine, delta-aminovaleric acid, and leucine.

When young children do not receive adequate amounts of the micronutrient iodine in their diet, their growth and cognitive development can be impaired. Nearly every country in the world has programs in place to track iodine intake and provide supplemental iodine if needed, usually in the form of fortified salt. The iodine nutrition status of a population can be tracked by monitoring iodine levels in urine samples to see if the median value falls in the range of 100–300 micrograms of iodine per liter of urine (μg I/L), which indicates adequate or more than adequate iodine nutrition. Many low and middle-income countries (LMIC) do not have a laboratory capable of carrying out this challenging assay, so samples must be sent out for assay in external labs, which is expensive and time-consuming. In most LMIC, population iodine surveys are carried out every 5–10 years, which limits the utility of the data for program monitoring and evaluation. To solve this problem, we developed a field-friendly paper test card that uses the Sandell-Kolthoff reaction to measure urinary iodine levels. A blind internal validation study showed that 93% of samples (n = 60) of iodide in an artificial urine matrix were categorized correctly by visual analysis as deficient, adequate, or excessive for levels set forth by the World Health Organization. Quantitative measurements based on computer image analysis had an error of 40 ± 20 μg I/L (n = 35 for samples in the calibration range) and these results categorized 88% of the samples (n = 60) correctly. We employed lifecycle analysis principles to address the known toxicity of arsenic, which is an obligatory reagent in the Sandell-Kolthoff reaction. Disposal of the cards in a landfill (their most likely destination after use) could let arsenic leach into groundwater; toxicity characteristic leaching procedure (TCLP) tests showed that the level of arsenic leached from the cards was 28.78 ppm, which is above the United States Environmental Protection Agency’s limit of 5 parts per million for solid waste. We integrated a remediation module into the card. This module contains oxone, to oxidize As(III) to As(V) oxyacids, and the iron oxide goethite. TCLP testing showed that the leachable amount of arsenic was reduced by at least 97.6%—from 28.8 ppm to lower than 0.7 ± 0.7 ppm (n = 20). This upstream intervention rendered the test card suitable for landfilling while retaining its functionality to perform a critical public health evaluation. [ABSTRACT FROM AUTHOR]

Blood filter paper strips are cost-effective materials used to store body fluid specimens under challenging field conditions, extending the reach of zoonotic pathogen surveillance and research. We describe an optimized procedure for the extraction of parasite DNA from whole blood (WB) stored on Type I Advantec Nobuto strips from both experimentally spiked and field-collected specimens from canine and skunks, respectively. When comparing two commercial kits for extraction, Qiagen's DNeasy Blood & Tissue Kit performed best for the detection of parasite DNA by PCR from Trypanosoma cruzi-spiked canine WB samples on Nobuto strips. To further optimize recovery of β-actin from field-collected skunk WB archived on Nobuto strips, we modified the extraction procedures for the Qiagen kit with a 90 °C incubation step and extended incubation post-addition of proteinase K, a method subsequently employed to identify a T. cruzi infection in one of the skunks. Using this optimized extraction method can efficaciously increase the accuracy and precision of future molecular epidemiologic investigations targeting neglected tropical diseases in field-collected WB specimens on filter strips. [ABSTRACT FROM AUTHOR]

Introduction and aim. In the literature, blood is described as a body fluid that circulates in a closed vascular system that makes up about 10% of the body weight. The morphotic elements of blood are suspended in this fluid, which is why we can define blood as liquid tissue. Blood in the organization helped to play a role, performing a transport, hemodynamic and protective-defensive function. Its main components are red blood cells, white blood cells and platelets. Erythrocytes due to the ability to connect to the source, participate in the delivery from the lungs to the source. Leukocytes include a group of blood cells that include monocytes, lymphocytes and granulocytes that participate in the body's reactions. Platelets are referred to in the literature as thrombocytes, as non-nucleated blood components. The main component of blood samples is participation in hemostatic processes in the composition. They play one key role in the blood clotting process. At some point, when the continuity of the straight line is interrupted, platelets pass to the subendothelial matrix, the formation of a platelet plug, a clot is formed. Until the beginning of the twentieth, blood functions are provided only for hemostatic activities. Recent scientific research that blood cells are involved in non-hemostatic processes. Including the immune system. Collecting information on the role of blood in an independent team, enabling review work. Material and methods. Literature search from 1989 up to 2022 has been conducted on PubMed, Google Scholar databases and the Practical Medicine for Physicians. For the final analysis, 10 research papers and 9 review papers were used. Following keywords were used to search articles: platelets, in- flammation, immune system, hemostasis. The articles were from the years 1989-2022. Results. Platelets are one of the main morphotic elements of blood, when it comes to size, they are the smallest of all components of peripheral blood, but they are very reactive. They participate in the processes of coagulation and maintaining hemostasis in the body. The characteristic properties of platelets are the presence of dense granules δ and α, their compactness during hemostasis or inflammation are released into the body. Platelets, in addition to active participation in hemostasis processes, take part in immune processes. Platelets have been shown to be regulators of immune processes through the presence of TLRs that trigger the initiation of an immune system response. In addition to the regulatory properties, platelets are involved in the innate immunity and inflammation of the body. Platelets with the participation of endothelial cells lead to the recruitment of leukocytes to the site of inflammation. Platelets participate in the recognition of a foreign pathogen that is in the body, the mechanisms that lead to this are presented in the paper. In addition to the function of platelets in the immune system, the topic of autoimmune diseases, where platelets play a role, is discussed. The work deals with the subject of the influence of platelets on the immune system and their role in immune processes. Through the progress of scientific research, platelets have been shown to play a significant role in immune mechanisms. The work is a review of literature dealing with topics related to the immune system and platelets and aims to explain the mechanisms that lead to the active participation of platelets in the immune system. Conclusion. Platelets are involved in the immune system, it has been shown at the same time that platelets perform regulatory functions in the immune system through the CD154 molecule and additionally participate in inflammation. Where are the first cells that are responsible for fighting in the body's non-specific immune response. In addition, TLR receptors that are found on the surface of platelets play a key role in recognizing a threat and initiating an immune response. Through these receptors, platelets recognize pathogens in the body. Scientists continue to study the non-haemostatic functions of platelets and discover their significant role in the body, which is not only based on blood coagulation processes, but to a large extent in the body's immune defense. [ABSTRACT FROM AUTHOR]

We developed an affordable, highly sensitive, and specific paper-based microfluidic platform for fast multiplexed detections of important biomarkers in various body fluids, including urine, saliva, serum, and whole blood. The sensor array consisted of five individual sensing channels with various functionalities that only required a micro liter-sized sample, which was equally split into aliquots by the built-in paper microfluidics. We achieved the individual functionalizations of various bioreceptors by employing the use of wax barriers and 'paper bridges' in an easy and low-cost manner. Pyrene carboxylic acid-modified single-walled carbon nanotubes (PCA/SWNTs) were deposited by quantitative inkjet printing with an optimal 3-dimensional semiconductor density on a paper substrate. Multiple antibodies were immobilized onto the SWNTs surface for highly sensitive and specific field-effect transistor (FET)/chemiresistor (CR) biosensors. We explored the optimal sensing conditions for the paper-based CR biosensor to achieve high sensitivities and specificities towards the target biomarker proteins (human serum albumin (HSA) and human immunoglobulin G (HIgG)) and achieved an ultralow detectable concentration of HSA and HIgG at 1.5 pM. Besides, origami folding was employed to simplify the fabrication process further. The sensing platform described in this work was cost-effective, semi-automated, and user-friendly. It demonstrated the capability of having multiple sensing functions in one paper-based microfluidic sensing platform. It envisioned the potential of a point-of-care device with full-analysis for practical diagnostics in an ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free and Deliverable to end-users) fashion for a quick test of targets of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Sweat, commonly referred to as the ultrafiltrate of blood plasma, is an essential physiological fluid in the human body. It contains a wide range of metabolites, electrolytes, and other biologically significant markers that are closely linked to human health. Compared to other bodily fluids, such as blood, sweat offers distinct advantages in terms of ease of collection and non-invasive detection. In recent years, considerable attention has been focused on wearable sweat sensors due to their potential for continuous monitoring of biomarkers. Electrochemical methods have been extensively used for in situ sweat biomarker analysis, as thoroughly reviewed by various researchers. This comprehensive review aims to provide an overview of recent advances in non-electrochemical methods for analyzing sweat, including colorimetric methods, fluorescence techniques, surface-enhanced Raman spectroscopy, and more. The review covers multiple aspects of non-electrochemical sweat analysis, encompassing sweat sampling methodologies, detection techniques, signal processing, and diverse applications. Furthermore, it highlights the current bottlenecks and challenges faced by non-electrochemical sensors, such as limitations and interference issues. Finally, the review concludes by offering insights into the prospects for non-electrochemical sensing technologies. By providing a valuable reference and inspiring researchers engaged in the field of sweat sensor development, this paper aspires to foster the creation of innovative and practical advancements in this domain. [ABSTRACT FROM AUTHOR]

Aim: To investigate if free PSA (fPSA) and total PSA (tPSA) values obtained from simultaneously collected urine, fresh and dried on filter paper, reflect the serum free and total PSA. Materials and Methods: The sera and 20 cc first voided urine from 33 consecutive men aged between 40 and 84 (mean 61 ± 12), were collected in the morning and delivered to the laboratory. Three different aliquots of 100 microgram urine were taken with automatic pipette and dropped on 3 certain areas of a filter paper and allowed to dry for each patient. On each paper, borders of dried urine were marked. PSA values were obtained from the sera and fresh urine samples and recorded. Later on particular days dried urine samples were dissolved and eventually PSA values were derived and recorded again. The results were compared to each other. Correlations were evaluated by using an SPSS statistics program. Results: Serum PSA values correlated weakly (r < 0.24) with fresh and dried urine PSA values. While PSA in fresh and dried urine samples showed strong correlation (0.5 < r < 0.74), a very strong correlation (r > 0.75) among PSA values of dried urine samples of 1-day, 7-.and 28-days, were seen. Conclusions: We conclude that PSA values obtained from fresh and dried urine could not reflect serum PSA values. But, because dried urine on a filter paper can be stable for years, it could be used for forensic purposes. [ABSTRACT FROM AUTHOR]

Quick and effective point-of-care (POC) devices have the chance to revolutionize healthcare in developed and developing countries since they can operate anywhere the patient is, with the possibility of obtaining and sending the results to the doctor without delay. In recent years, significant efforts have focused on developing new POC systems that can screen for biomarkers continuously and non-invasively in body fluids to prevent, diagnose, and manage diseases. However, one of the critical challenges left to address is how to power them effectively and sufficiently. In developing countries and rural and remote areas, where there are usually no well-established electricity grids or nearby medical facilities, and using batteries is unreliable or not cost-effective, alternative power sources are the most challenging issue for stand-alone and self-sustained POC devices. Here, we provide an overview of the techniques for used self-powering POC devices, where the sample is used to detect and simultaneously generate energy to power the system. Likewise, this paper introduced the state-of-the-art with a review of different research projects, patents, and commercial products for self-powered POCs from the mid-2010s until present day. [ABSTRACT FROM AUTHOR]