High Reproducibility Research Articles

Development and optimization of multivesicular gefitinib liposomal transdermal system employing lipoid S100 for breast cancer: pharmacokinetics, bioavailability, and skin irritation studies in Wistar rats

BackgroundConventional therapies in cancer treatment face challenges including drug resistance, lack of specificity, and severe adverse reactions. This study explores the potential of liposomal transdermal delivery systems as an alternative to current therapies with improved BA and PK. The objective of the study was to formulate gefitinib liposomes by thin film hydration technique (TFH) using lipoid S100. A central composite design (CCD) was used to develop and optimize GEF-LIP-TDDs and to analyze the optimum concentrations of the selected variables (phospholipid, cholesterol) in liposomal formation. The model fitting was performed using Design-Expert (Stat-Ease, Ver 13). The GEF liposomes were evaluated for %EE, mean particle size and PDI. The optimized liposomes were fabricated as a transdermal patch by mercury substrate method and evaluated for %drug content, in vitro diffusion, in vivo biodistribution (PK and BA), and skin irritation studies in female Albino Wistar rats. The stability of the optimized transdermal patch was also assessed for 3 months.ResultsThe CCD model was significant with F-value of 37.97, P-value of 0.0500 and R2 of 0.9644. The average vesicle size, PDI, and ZP of GEF-LIPs (F1–F13) were found to be between 112.8 to 373.7 nm, 0.186 to 0.510 and − 3.69 to − 82.2 mV, respectively. F3-GEF-LIP exhibited a mean vesicle size of 96.07 nm, ZP of − 46.06 mV, and a PDI of 0.423. F3-GEF-LIP demonstrated exceptional %EE (97.79) and sustained release effect (%CDR, 83.32) following a diffusion-controlled mechanism. TEM images confirmed liposomes of multivesicular type (MVV, < 100 nm). Importantly, optimized F3-GEF-LIP-TD showed no signs of edema in Wistar rats. The biodistribution of F3-GEF-LIP-TD was similar to pure GEF and was higher in the liver (p < 0.05). The BA of F3-GEF-LIP-TD was observed to be 74.05 ± 0.11% in comparison with oral GEF-LIP (65.25 ± 0.08%) and pure GEF (58.10 ± 0.17%).ConclusionTFH technique offers stable liposomes with high reproducibility. Our findings imply that GEF-LIP-TD provides enhanced BA and tissue distribution and can be considered as a substitution for orals or in combination for treating breast cancer. Lipoid S100 is a potential lipid for developing stable multivesicular nanoliposomes.Graphical abstract

Interlaboratory comparison of standardised metabolomics and lipidomics analyses in human and rodent blood using the MxP® Quant 500 kit.

Metabolomics and lipidomics are pivotal in understanding phenotypic variations beyond genomics. However, quantification and comparability of mass spectrometry (MS)-derived data are challenging. Standardised assays can enhance data comparability, enabling applications in multi-center epidemiological and clinical studies. Here we evaluated the performance and reproducibility of the MxP® Quant 500 kit across 14 laboratories. The kit allows quantification of 634 different metabolites from 26 compound classes using triple quadrupole MS. Each laboratory analysed twelve samples, including human plasma and serum, lipaemic plasma, NIST SRM 1950, and mouse and rat plasma, in triplicates. 505 out of the 634 metabolites were measurable above the limit of detection in all laboratories, while eight metabolites were undetectable in our study. Out of the 505 metabolites, 412 were observed in both human and rodent samples. Overall, the kit exhibited high reproducibility with a median coefficient of variation (CV) of 14.3 %. CVs in NIST SRM 1950 reference plasma were below 25 % and 10 % for 494 and 138 metabolites, respectively. To facilitate further inspection of reproducibility for any compound, we provide detailed results from the in-depth evaluation of reproducibility across concentration ranges using Deming regression. Interlaboratory reproducibility was similar across sample types, with some species-, matrix-, and phenotype-specific differences due to variations in concentration ranges. Comparisons with previous studies on the performance of MS-based kits (including the AbsoluteIDQ p180 and the Lipidyzer) revealed good concordance of reproducibility results and measured absolute concentrations in NIST SRM 1950 for most metabolites, making the MxP® Quant 500 kit a relevant tool to apply metabolomics and lipidomics in multi-center studies.

Chitosan-Based Electronics: The Importance of Acid Strength and Plasticizing Additives on Device Performance.

A rise in demand for disposable consumer electronics such as smart packaging, wearable electronics, and single-use point-of-source sensors requires the development of eco-friendly and compostable electronic materials. Chitosan is derived from crustacean waste and offers high dielectric constant values without requiring rigorous purification, making it sustainable for large-scale electronic device manufacturing. When processed in acidic media, the protonated backbone of chitosan pairs with counterions from the acid dissociation to form chitosan thin films with electrical double layers (EDLs) and tunable capacitive properties. We report the importance of the choice of acid when processing chitosan by surveying a series of halogenated and biosourced acids with varying pKa values and solutions with different pH values. Oxalic acid outperforms other acids, with a maximum areal capacitance of 161 nF·mm-2. Tartaric acid and citric acid, despite lower capacitance values, showed promising results with a stable EDL capacitance and high reproducibility, making them optimal for large-area manufacturing. The incorporation of sorbitol as a plasticizer boosts the EDL formation onset of all chitosan-acid combinations to 1 × 103-105 Hz and improves reproducibility. High-performing single-walled carbon nanotube thin film transistors were made using chitosan-based dielectrics treated with different acids with and without sorbitol, leading to transconductance as high as ≈5.2 μS and Ion/Ioff of 105. The capacitors and transistors remain functional after one year of storage in ambient conditions. Overall, this study demonstrates durable high-performance electronics based on chitosan and stresses the importance of processing acid and the use of plasticizing additives, such as sorbitol.

Glial Fibrillary Acidic Protein's Usefulness as an Astrocyte Biomarker Using the Fully Automated LUMIPULSE® System.

Glial fibrillary acidic protein (GFAP) is an important biomarker for neuroinflammatory conditions. Recently, advancements in the treatment of neurological diseases have highlighted the increasing importance of biomarkers, creating a demand for accurate and simple measurement systems for GFAP levels, which are essential for both research and clinical applications. This study presents the development and validation of a novel fully automated immunoassay for the quantitative determination of GFAP levels in biological samples. We examined the analytical performance of the GFAP assay on the LUMIPULSE platform. The assay's parameters, including antibody concentrations, incubation times, and detection methods, were optimized to enhance sensitivity and specificity. GFAP levels were measured in 396 serum or plasma samples, comprising both healthy controls and patients with neurodegenerative diseases. In the analytical performance studies, intra- and inter-assay coefficients of variation (CV) were below 5%, indicating high reproducibility. Additionally, the assay demonstrated good linearity over the measurement range. The limit of quantification (LoQ) for this assay was 6.0 pg/mL, which is sufficient for measuring specimens from healthy controls. In clinical validation studies, GFAP levels were significantly elevated in patients with neurodegenerative diseases compared to healthy controls. This automated GFAP assay provides a robust and reliable tool for GFAP measurement, facilitating further research into GFAP's role in neurological disorders and potentially aiding in the diagnosis and monitoring of these conditions.

NIMG-26. GLIOBLASTOMA TISSUE INVASION MAPPING WITH NOVEL HYPOXIA-TARGETED BLOOD OXYGENATION-LEVEL DEPENDENT MRI

Abstract BACKGROUND Glioblastoma is a devastating incurable malignant primary brain tumor exhibiting an erratic growth pattern and brain tissue invasion exceeding the typically depicted contrast-enhancing lesion on MRI. This behaviour is strongly associated with the presence of aberrant vasculature. Advanced clinical neuroimaging techniques, however, have not provided convincing imaging markers to determine glioblastoma tissue infiltration and monitor treatment effects. We investigated a new imaging technique based on blood oxygenation-level dependent (BOLD) MRI contrast with precise hypoxic respiratory targeting in order to better determine tumor tissue invasion not restricted to the typically depicted focal glioblastoma lesion on contrast-enhanced MRI. METHODS A computer-controlled gas blender was used to induce transient and standardized hypoxic stimulus in isocapnic conditions during echo-planar-imaging acquisition in patients with brain tumors.8,9 Data were preprocessed using SPM and in-house written MATLAB scripts. The induced BOLD signal changes were used to calculate %BOLD signal change during hypoxic stimulus with respect to the baseline, contrast to noise (CNR) ratio and goodness-of-fit (Rsquared) for each voxel in the whole-brain and automatically segmented region of interest (contrast-enhancement, necrosis, edema) and overlayed using color-maps onto anatomical high resolution T1. RESULTS Hypoxic targeting during gas modulation could be performed with high reproducibility, caused no discomfort in patients with brain tumors and was able to induce strong signal change during hypoxic stimulus with high contrast-noise ratio and Rsquared. In particular, when compared with other tumors, BOLD hypoxia glioblastoma tissue invasion maps show very distinct hypoxic signal responses in some cases extending beyond the focal -contrast-enhancing- glioblastoma lesion. CONCLUSION The ability to map at voxel level BOLD hypoxia signal patterns may determine a novel therapeutic imaging marker based on structural vascular alterations in the tumour tissue infiltration zone before these become appreciable with standard neuroimaging. This may support a refinement and tailoring of tumor resection and improve follow-up after resection and during treatment i.e. chemoradiotherapy.

Age-Related Differences and Reliability of a Field-Based Fitness Test Battery in Young Trained Footballers: The Role of Biological Age.

the purpose of this study was to analyze the reliability of a field-based fitness test battery in young trained football players, according to biological age. 197 young trained football players (12-19 years old) participated in the study. We measured anthropometric measurements (i.e., height, sitting height, length leg, and body mass), a bilateral vertical jumping test (CMJ), a progressive loading test of squats and hip thrust, acceleration and speed tests (10 m and 30 m sprint tests), a change-of-direction ability test (V-cut test), and a cardiorespiratory fitness test (30-15 intermittent fitness test). Statistical data are shown as the mean ± standard deviation by PHV group in tests and retests. Test reliability was assessed through the intraclass correlation coefficient (ICC), with an ICC above 0.9 being considered high. To evaluate accuracy and repeatability, standard error of measurement, coefficient of variation, and minimum detectable change at 90% were determined and Bland-Altman diagrams were used, establishing a statistical significance of p < 0.05. All of the tests showed non-significant differences between the test and retest in the pooled sample (p > 0.05). Furthermore, all of them presented a trivial effect size (<0.2) and high intraclass correlation coefficients (>0.9), which indicates the high reproducibility of the tests, despite some of them presenting a significant difference between trials (i.e., the CMJ, 10 m sprint, V-cut, and squat tests). Low measurement errors were found for all tests (coefficient of variation [CV] = 4.39-9.39), except for the CMJ and the progressive loading test for squat and hip thrust exercises (CV = 12.2-21.11). Similar results were found irrespective of biological age group. All tests were reliable for the pooled sample as well as for the biological age groups.

Electrochemical C−O and C−N Arylation using Alternating Polarity in flow for Compound Libraries

AbstractEtherification and amination of aryl halide scaffolds are commonly used reactions in parallel medicinal chemistry to rapidly scan structure–activity relationships with abundant building blocks. Electrochemical methods for aryl etherification and amination demonstrate broad functional group tolerance and extended nucleophile scope compared to traditional methods. Nevertheless, there is a need for robust and scale‐transferable workflows for electrochemical compound library synthesis. Herein we describe a platform for automated electrochemical synthesis of C−X arylation (X=NH, OH) in flow to access compound libraries. A comprehensive Design of Experiment (DoE) study identifies an optimal protocol which generates high yields across&gt;30 aryl halide scaffolds, diverse amines (including electron‐deficient sulfonamides, sulfoximines, amides, and anilines) and alcohols (including serine residues within peptides). Reaction sequences are automated on commercially available equipment to generate libraries of anilines and aryl ethers. The unprecedented application of potentiostatic alternating polarity in flow is essential to avoid accumulating electrode passivation. Moreover, it enables reactions to be performed in air, without supporting electrolyte and with high reproducibility over consecutive runs. Our method represents a powerful means to rapidly generate nucleophile independent C−X arylation compound libraries using flow electrochemistry.

Eco-friendly synthesis and surface modification of ZnO nanoparticles using Pueraria montana root extract: enhanced photocatalytic performance with trisodium pyrophosphate

ABSTRACT The synthesis of zinc oxide (ZnO) nanoparticles using Pueraria montana (kudzu) plant extracts highlights advancements in green nanotechnology. This study explores the use of Pueraria montana roots, rich in phytochemicals such as phenols, terpenoids and flavonoids, which serve as natural reducing and stabilizing agents for nanoparticle synthesis. The eco-friendly production and surface modification of ZnO nanoparticles were achieved using these plant extracts, enhancing their photocatalytic performance with trisodium pyrophosphate (TSPP).Characterization techniques, including XRD, TEM, and FE-SEM, confirmed the nanoparticles’ crystalline structure, with a BET surface area of 14.01 m²/g and a type II adsorption isotherm. The synthesized nanoparticles exhibited exceptional dye degradation capabilities, achieving 99.9% removal of malachite green (MG) and 97.78% for aniline blue (AB). Kinetic analysis revealed a steady-state rate constant (k) of 2 × 10⁻⁴ min⁻¹ for MG where as the response rate constant(k) for the photocatalytic degradation of AB using modified ZnO nanostructures was 3.0 × 10−3 min−1 allowing near-total dye removal within 600 minutes for MG and 350 min for AB . Adsorption studies were well described by Langmuir and Freundlich isotherms, indicating maximum adsorption capacities of 59.7 mg/g for MG and 61.2 mg/g for AB. Surface modification with TSPP improved charge carrier separation and enhanced the generation of reactive oxygen species under UV exposure, further increasing dye degradation efficiency.Statistical analysis indicated high reproducibility, with close mean adsorption values of 99.125 for MG and 97.3125 for AB, supported by small standard deviations. The findings affirm the potential of using Pueraria montana in synthesizing efficient photocatalysts, providing a sustainable and cost-effective approach for water treatment applications. Overall, this research underscores the effectiveness of plant-based materials in advancing eco-friendly nanotechnology solutions.

Construction of a portable and sensitive electrochemical immunosensor for the rapid detection of erythromycin based on semiconductive bimetallic MOF

The sensitive determination of antibiotics in food products is vital for ensuring food safety and protecting human health. Herein, we have fabricated a novel electrochemical portable and sensitive electrochemical immunosensor for the efficient detection of erythromycin (ERY) containing in food stuffs. For this, a semiconductive cooper/ferric bimetallic metal-organic framework (scMOF), which was synthesized using 2,3,6,7,10,11-hexahydroxytriphenylene (HHTP) as linking ligand (denoted as CuxFe3-x(HHTP)2), was utilized simultaneously as the platform for anchoring antibody and for modifying the sprinted printed electrode (SPE) to construct the electrochemical immunosensor. The obtained scMOF, CuxFe3-x(HHTP)2, exhibited high porosity, promoted conductivity, and enhanced anchoring ability toward antibody. Thereby, the developed SPE immunosensor demonstrated the superior biosensing performance for the detection of ERY. Within a wide range from 1.0 fg mL−1 to 1.0 ng mL−1, the CuxFe3-x(HHTP)2-based portable SPE immunosensor had an ultralow detection limit of 0.69 fg mL−1, together with high selectivity, good reproducibility, and excellent long-term stability, as well as acceptable practicality. The present SPE immunosensor based on scMOFs not only provides an innovative biosensing strategy for the sensitive inspection of antibiotics, but also extends the application of scMOF in the field of food analysis.

Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs.

The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 109-6.43 × 101 copies/µL, with an R2 value of 0.996 and a standard curve equation of y = -3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings.

Photonic sensor based on surface imprinted polymers for enhanced point-of-care diagnosis of bacterial urinary tract infections

Effective bacterial detection is crucial for health diagnostics, particularly for the detection of pathogenic species like Escherichia coli (E. coli), which is responsible for up to 90% of urinary tract infections (UTIs), is especially crucial. Current detection methods are time-consuming, often delaying diagnosis and treatment. This study introduces an innovative approach for rapid E. coli detection using porous silicon (PSi) substrates combined with Surface Imprinted Polymers (SIPs) for photoluminescence-based (PL-based) E. coli detection. The PSi/SIP substrates offer high sensitivity, selectivity, and a low limit of detection (LOD) without the need for natural recognition elements. These substrates, fabricated via metal-assisted chemical etching (MACE) and PDMS-based E. coli imprinting, demonstrate reliable repeatability and a fast detection. Real-time detection experiments in phosphate-buffered saline (PBS) and urine showed consistent stair-like quenching of the PL signal with increasing E. coli concentrations, achieving theoretical LODs of approximately 13 ± 2 CFU/mL in PBS and 17 ± 3 CFU/mL in urine. The substrates exhibited excellent selectivity, differentiating E. coli from other species such as Cronobacter sakazakii (C. sakazakii) and Listeria monocytogenes. The high sensitivity and reproducibility of PSi/SIP substrates, along with the ease of use and rapid detection capabilities of the resulting sensor, highlight the potential of this novel platform for point-of-care (PoC) applications in clinical diagnostics.

Femtosecond laser-inscribed in-fiber Mach-Zehnder interferometer for ultra-sensitive small-angle torsion measurement

We propose and demonstrate a novel in-fiber Mach-Zehnder interferometer (MZI), using a femtosecond (FS) laser to inscribe two parallel straight waveguides along both sides of the core-cladding boundary of an optical fiber. Both simulations and experiments demonstrated that constructing a second straight waveguide (WG2) on the opposite side of the fiber core significantly enhances the mode coupling efficiency in fiber, resulting in ultra-sensitive small twist-angle torsion measurement by intensity interrogation. By finely adjusting the waveguide spacing between the WG2 and the fiber core during the FS fabrication process, a large tuning range of ∼ 200 nm for the designable interference wavelength with maximal fringe contrast has been achieved. Through comparative experiments, the proposed in-fiber MZI performs ultra-sensitivity with 17417.92 dB/(rad/mm) over small twist angles from −10° to 0° for the sample “WG1-4/WG2-4” and 19480.57 dB/(rad/mm) over small twist angles from 0° to 10° for the sample “WG1-4/WG2-6”. Besides, the device is insensitive to temperature and strain. The exhibited advantages of high sensitivity, high reproducibility, and low crosstalks promote the proposed in-fiber MZI to be applied in future industrial engineering monitoring and intelligent health monitoring.

Highly efficient electrocatalysts of NiMoO4 doped CdO nanostructure interconnected with modified glassy carbon electrode for the detection of chloramphenicol

In this study, we developed an excellent electrocatalyst of NiMoO4 doped with CdO and then coated electrochemical detection of chloramphenicol (CAP) via a glassy carbon electrode (NiMoO4@CdO/GCE). In particular, this drug treats typhoid, fever, and salmonellosis infections. CAP detection and measurements have become progressively important in ensuring human safety. The NiMoO4@CdO nanocomposite was prepared using the one-step hydrothermal method and characterized for its structural and optical properties. The electrochemical determination of CAP was investigated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) techniques. We observed that the glassy carbon electrode modified with NiMoO4@CdO efficiently reduced the CAP at a very low potential. While CAP interacts with this system, metal oxide nanocomposites can combine with CAP reduction to produce phenylhydroxylamine. According to the DPV analysis, the bare electrode provides a linear response in the 10–100 µM range and a substantial reduction in CAP over a low detection limit of 0.083 µM when compared to (NiMoO4@CdO/GCE). Other co-existing molecules challenge the utility of this sensor. Additionally, this sensor exhibited high selectivity, stability, and reproducibility. Further, we evaluated the sensor for real-sample analysis, and the results were satisfactory.

Tetraphenylethene-Containing Nanofilm via Interfacial Confinement Self-Assembly for Fluorescent Monitoring of SOCl2 Leakage in a Li-SOCl2 Battery Electrolyte.

Lithium thionyl chloride (Li-SOCl2) batteries are widely used due to their high energy density and long shelf life. However, the corrosive nature of SOCl2 poses a safety hazard, necessitating effective leak detection methods. We report an approach for real-time fluorescent detection of SOCl2 leakage in Li-SOCl2 batteries using a tetraphenylethene-based nanofilm. The nanofilms were prepared through the interfacial confined self-assembly method and exhibit excellent flexibility, homogeneity, tunable size and thickness, and adaptability to various substrates, enabling easy integration into sensor devices. They possess high photochemical stability and a photoluminescence quantum yield exceeding 25%, demonstrating their potential as high-performance fluorescent sensing material. The nanofilms also exhibit high sensitivity, good reproducibility, and selectivity toward SOCl2 detection. Upon exposure to SOCl2 vapor, the nanofilm shows a red-shifted and fluorescence quenching response, attributed to the protonation of the acylhydrazone bond in the nanofilm by the hydrolysis product of SOCl2, which disrupts the electronic structure of the nanofilm and leads to a decrease in the fluorescence intensity and a shift in the emission wavelength. A detection limit of ∼1.31 ppt and excellent repeatability over 300 cycles are demonstrated, highlighting their high sensitivity and reliability. This work paves the way for a highly sensitive and reliable leak detection system for Li-SOCl2 batteries, enhancing their safety and reliability in various applications.

Infrared Laser Ablation and Capture of Formalin-Fixed Paraffin-Embedded Tissue.

Formalin-fixed paraffin-embedded (FFPE) tissue is a ubiquitous and invaluable resource for biomedical research and clinical applications. However, FFPE tissue proteomics is challenging due to protein cross-linking and chemical modification. Laser ablation sampling allows precise removal of material from tissue sections with high spatial control and reproducibility for offline proteomics by liquid chromatography coupled with tandem mass spectrometry. In this work, we used a pulsed mid-infrared laser for microsampling of rat liver tissue for subsequent identification and quantification of proteins. It was found that more proteins were identified by FFPE tissue laser ablation sampling compared to fresh frozen (FF) tissue laser ablation sampling and that more proteins were identified by laser ablation than by manual dissection of FFPE tissue. In contrast to previous studies, no loss of hydrophilic proteins due to residual cross-linking was observed. The efficient capture of proteins by laser ablation microsampling is attributed to efficient laser breakup of the tissue which facilitates downstream processing of the proteins.

Clinical validation of an AI-based pathology tool for scoring of metabolic dysfunction-associated steatohepatitis.

Metabolic dysfunction-associated steatohepatitis (MASH) is a major cause of liver-related morbidity and mortality, yet treatment options are limited. Manual scoring of liver biopsies, currently the gold standard for clinical trial enrollment and endpoint assessment, suffers from high reader variability. This study represents the most comprehensive multisite analytical and clinical validation of an artificial intelligence (AI)-based pathology system, AI-based measurement of metabolic dysfunction-associated steatohepatitis (AIM-MASH), to assist pathologists in MASH trial histology scoring. AIM-MASH demonstrated high repeatability and reproducibility compared to manual scoring. AIM-MASH-assisted reads by expert MASH pathologists were superior to unassisted reads in accurately assessing inflammation, ballooning, MAS ≥ 4 with ≥1 in each score category and MASH resolution, while maintaining non-inferiority in steatosis and fibrosis assessment. These findings suggest that AIM-MASH could mitigate reader variability, providing a more reliable assessment of therapeutics in MASH clinical trials.

Mechanisms of lipid-mediated regulation of the pore-forming activity of antimicrobial agents: studies on planar lipid bilayers

Planar lipid bilayers are unique tools designed for modeling cell membranes and electrophysiological studies of incorporated ion channels. Such model systems are designed to limit the number of components taking part in the functioning of biological membranes in order to characterize in detail the occurring processes under well-controlled experimental conditions. Planar lipid bilayers make it possible to record single events with a measured current of more than a tenth of a picoampere. The relative simplicity of the method, the ability to observe single molecular events, and the high reproducibility of the results obtained determines the unprecedented effectiveness of using planar lipid bilayers to identify key physical and chemical factors responsible for regulating the functioning of ion channels. This review represents an analysis of literature data concerning the mechanisms of lipid-associated regulation of ion channels formed by various antimicrobial agents. The examination allows us to consider the lipids as molecular chaperones that ensure the formation of pores in target membranes by antimicrobials.

Review of domestic test systems for detecting current or past SARS‑CoV‑2 infection using enzyme‑linked immunosorbent assay

Introduction. Rapid and reliable assessment of circulating antibody levels to SARS-CoV-2 remains the current challenge for monitoring and preventing the spread of COVID-19. As the risk of emergence and dissemination of new viral strains continues to grow, epidemiologic research is also becomes increasingly important. The enzyme-linked immunosorbent assay (ELISA) is useful to assess the vaccination effectiveness and the need for revaccination, the proportion of post-disease and vaccinated individuals, and to monitor population immunity. This article provides an overview of the local ELISA kits currently available for the detection of antibodies against SARS-CoV-2 viral proteins, with a comparison to the certified kit by Mediagnost Ltd. (Germany).The aim. Laboratory testing of seven domestic reagent kits for the determination of antibodies to SARS-CoV-2 viral proteins by ELISA in comparison with the certified test system “Anti-SARS-CoV-2 ELISA E 111-IVD” (Mediagnost Ltd., Reutlingen, Germany).Materials and methods. The study included 80 serum samples from conditionally healthy donors: experimental group (n=70, serum samples collected after 2019) and control group (n=10, serum samples collected before 2018). The experimental group is comprised of non-infected and post-disease individuals; a proportion of those were vaccinated against COVID-19.Results and conclusions. The data obtained indicate high reproducibility, sensitivity (100 %), and specificity for all domestic ELISA kits in detecting both post-disease (90–95 %) and vaccinated volunteers (100 %). The main quality criteria of the obtained results were comparable to those for the certified test system, “Anti-SARS-CoV-2 ELISA E 111-IVD” (Mediagnost Ltd., Germany). All kits detecting IgG to the receptor-binding domain (RBD) and to the S-glycoprotein were effective for the reliable assessment of antibody production after the vaccination. Kits for the detection of antibodies to the RBD and/or viral nucleocapsid are useful for identifying unvaccinated post-disease individuals.

A novel label-free impedance biosensor for KRAS G12C mutations detection based on PET-RAFT and ROP synergistic signal amplification

The KRAS G12C mutations, as crucial biomarkers, are closely associated with non-small cell lung cancer. Here, a novel label-free electrochemical biosensor with synergistic signal amplification of photocell energy transfer-reversible addition fragmentation chain transfer (PET-RAFT) and ring-opening polymerization (ROP) was developed for the first time for sensitive detection of KRAS G12C mutations. Specifically, hairpin DNA (hDNA), which act as biomolecular probe, was self-assembled on Au electrode surface by Au-S bond. 4-cyano-4-[(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid (CDTPA), the chain transfer agent of PET-RAFT reaction, was then attached to hDNA via amide bond. After that, the target DNA (tDNA) was captured on the electrode surface by complementary base pairing with hDNA. Subsequently, large numbers of electro-active monomers N-acryloxysuccinimide (NAS) were successfully grafted to the electrode surface via PET-RAFT reaction, which provided plenty of junction sites for doxorubicin-polycaprolactone (Dox-PCL) synthesized by ROP. Finally, the Dox-PCL was connected to the electrode surface by ester bond, significantly amplifying the electrochemical signal. Under optimized conditions, the biosensor has a wide linear detection range of 0.1 pM to 1 μM, with a detection limit of 86.9 fM. Attribute to its high sensitivity, specificity, reproducibility and stability, this biosensor possesses considerable potential in early diagnosis of disease and biomedical research.

Cross-cultural Adaptation of the Iranian Version of the Voice Symptom Scale

Background and Objectives: This research aims to evaluate the validity and reliability of the Persian version of the voice symptom scale (VoiSS) questionnaire. We also sought to determine the best cut-off point to classify high-risk populations into controls (participants) and dysphonic patients. Methods: The study was conducted in 3 main steps. The translation process was carried out by a pair of Iranian bilingual speech-language pathologists along with an expert panel. For the validity study, the questionnaire was administered to 268 cases that were suspected of dysphonia. Finally, to assess the reliability, the questionnaire was completed twice in 2 weeks by 40 dysphonia patients. To normalize data generation 107 individuals without dysphonia completed the P-VoiSS, after that the cutoff value of the P-VoiSS was calculated. Results: Cronbach’s α was estimated at 0.914, and for impairment, emotional, and physical domains, it was 0.877, 0.926, and 0.725, respectively. We also estimated intra-cluster correlation (ICC) of 0.984 indicating high reproducibility of the Persian VoiSS questionnaire. The estimated intra-cluster correlation (ICC) for subscale was 0.962 for impairment, 0.989 for emotional, and 0.952 for physical domains. The Mean±SD score of the questionnaire for the healthy group was 10.1±5.9, while it was statistically higher in dysphonia patients (44.1±20.6). The statistical difference was observed in the subscales of the questionnaire between healthy and dysphonia groups (P&lt;0.05). The exploratory factor analysis determined that the Persian VoiSS has three subtest/latent factors like the original form. We also estimate the best cut-point at 20.5. Conclusion: The Persian adaptation of VoiSS is a dependable and effective tool that can be utilized for screening high-risk populations.