Body Fluids Research Articles

The Role of Surface Energy and Wettability in Polymer-Based Drug Delivery Systems: Enhancing Bioadhesion and Drug Release Efficiency

Polymer-based drug delivery systems are pivotal in advancing the precision and efficacy of modern therapeutics. Among the key factors influencing these systems, surface energy and wettability are crucial in determining their interaction with biological tissues and drug release behavior. Surface energy, which governs the adhesive properties of the polymers, plays a significant role in bioadhesion—a critical aspect for drug delivery routes that require prolonged contact with biological surfaces, such as mucosal membranes. Wettability, the extent to which a polymer surface can be wetted by biological fluids, influences both drug loading efficiency and release kinetics. Hydrophilic polymers, characterized by high wettability, typically facilitate rapid drug release, whereas hydrophobic polymers, with low wettability, offer controlled, prolonged release profiles. Additionally, the balance between hydrophilicity and hydrophobicity is essential for optimizing drug-polymer interactions, particularly in systems designed for sustained or targeted drug delivery. Advances in surface modification techniques have enabled precise control over these parameters, allowing for the customization of polymers to meet specific therapeutic requirements. This review discusses the interplay between surface energy, wettability, and drug delivery performance, highlighting how these properties have been manipulated to enhance bioadhesion, drug retention, and release profiles. The review also addresses the challenges in designing polymer wettability, such as maintaining stability, scalability, and regulatory compliance. Finally, it explores emerging trends, including stimuli-responsive polymers and nanotechnology integration, which hold promise for the next generation of drug delivery systems that are more efficient, targeted, and patient-specific, than current systems.

Innovative voltammetric techniques for bumadizone analysis in pharmaceutical and biological samples: emphasizing green, white, and blue analytical approaches

There are no documented electroanalytical methods for quantifying the anti-inflammatory drug bumadizone (BUM) in pharmaceutical or biological matrices. So, a new voltammetric method was developed to determine BUM at nano concentrations in pharmaceutical forms, in the presence of its alkaline degradant, and in biological fluids. Five electrodes were tested, including three nano-reduced graphene oxide (nRGO) electrodes (5%, 15%, and 20%), a carbon paste electrode (CPE), and a 10% nRGO-modified CPE. The 10% nRGO-modified electrode showed the best performance, offering high selectivity and low detection limits, with good linearity in the concentration range of 0.9 × 102 to 15 × 102 ng mL−1. Differential pulse voltammetry successfully applied this electrode for BUM determination in various samples, achieving excellent recovery without preliminary separation. The method was validated according to ICH guidelines and compared favorably to the reference method. Its environmental impact was assessed using AGREE and Eco-scale metrics in addition to the RGB algorithm, showing superior greenness and whiteness profiles due to safer solvents and lower energy consumption, along with high practical effectiveness using the BAGI metric.

A Luminescent MOF-Based Sensor for Monitoring of an Anticancer Drug and a Pyrethroid Fungicide Biomarker in Wastewater and Biological Fluids.

The extensive use of insecticides, such as pyrethroids, and pharmaceutical drugs, such as doxorubicin (DOX) has significantly increased to meet the growing demand for food production and disease treatment. Among them, 3-phenoxybenzoic acid (3-PBA), a metabolite of pyrethroid insecticides, poses various health and environmental risks. Similarly, DOX is a well-known anticancer drug and has been continuously used for many years. The high demand and unregulated disposal of these substances raise concerns for both humans and the environment. To address this issue, there is a pressing need to monitor the presence of these analytes in wastewater to protect our ecosystems. This challenge has inspired us to develop an MOF-based fluorometric dual sensor capable of rapid and selective detection of these analytes in aqueous solutions. This work represents the first MOF-based dual probe for detecting these targeted analytes. There was a 98% fluorescence quenching upon the introduction of DOX whereas about a 11-fold increment of the probe's fluorescence intensity took place in the presence of 3-PBA. The sensitivity of the probe is notably high as limits of detection (LOD) are 8.7 nM for DOX and 1.2 nM for 3-PBA. Our designed probe has the highest KSV value for DOX which is 3.37 × 106 M-1. The MOF demonstrated remarkable rapid response time of just 5 and 10 s for DOX and 3-PBA, respectively. The MOF exhibited outstanding selectivity in detecting DOX and 3-PBA, even when other interfering substances were present. We tested the probe's sensing abilities in various environments, such as serum, urine, wastewater, and different pH levels. These findings underscore the sensor's practicality and usefulness in real-world applications. The underlying mechanisms driving the sensing processes were thoroughly investigated by using various modern analytical methods.

Altered circadian expression of clock genes and clock-regulatory epigenetic modifiers in saliva of children with fetal alcohol spectrum disorders

Prenatal alcohol-exposed (AE) infants and children often demonstrate disrupted sleep patterns, including more frequent awakenings, reduced total sleep time, and more night-to-night sleep variability. Despite the strong connection between sleep patterns and circadian rhythmicity, relatively little is known about circadian rhythm disruptions in individuals with AE. Recently, several reports demonstrated that evaluating the expression patterns of human clock genes in biological fluids could reveal an individual’s circadian phenotype. Human saliva offers an emerging and easily available physiological sample that can be collected non-invasively for core-clock gene transcript analyses. We compared the expression patterns of core-clock genes and their regulatory genes in salivary samples of children aged 6–10 years-old with and without AE during the light cycle between ZT0-ZT11. We isolated the RNA from the samples and measured the expression patterns of core clock genes and clock regulating genes using the human specific primers with quantitative real-time PCR. Analysis of core clock genes expression levels in saliva samples from AE children indicates significantly altered levels in expression of core-clock BMAL1, CLOCK, PER1-3 and CRY1,2, as compared to those in age-matched control children. We did not find any sex difference in levels of clock genes in AE and control groups. Cosinor analysis was used to evaluate the rhythmic pattern of these clock genes, which identified circadian patterns in the levels of core clock genes in the control group but absent in the AE group. The gene expression profile of a salivary circadian biomarker ARRB1 was rhythmic in saliva of control children but was arhythmic in AE children. Altered expression patterns were also observed in clock regulatory genes: NPAS2, NFL3, NR1D1, DEC1, DEC2, and DBP, as well as chromatin modifiers: MLL1, P300, SIRT1, EZH2, HDAC3, and ZR1D1, known to maintain rhythmic expression of core-clock genes. Overall, these findings provide the first evidence that AE disturbs the circadian patten expression of core clock genes and clock-regulatory chromatin modifiers in saliva.

Surface-enhanced Raman spectroscopy in forensic analysis

Abstract Surface-enhanced Raman spectroscopy (SERS) is a modern analytical technique that can be used to determine the structure and composition of analyzed specimens at the molecular level. This technique is also minimally invasive and destructive. This and other advantages of SERS against traditional techniques used in forensics make it the ideal method for the identification of bodily fluids, soil constituents, drugs and chemicals, and colorants on or in hair, fabric, and paints. In this article, we first introduce the reader to the fundamental physical properties of SERS. Next, we discuss the most recent findings that are focused on SERS-based analyses of physical evidence related to forensics research. Finally, we provide a critical overview of where the future of SERS should be in forensics science.

Versatile, reusable and highly sensitive SERS-based point-of-care testing microplatform for reliable ATP detection

The advancement in miniaturized Raman spectrometers, coupled with the single-molecule-level sensitivity and unique fingerprint identification capability of surface-enhanced Raman scattering (SERS), offers great potential for point-of-care testing (POCT). Despite this, accurately quantifying analyte molecules, particularly in complex samples with limited sample volumes, remains difficult. Herein, we present a versatile and reusable SERS microplatform for highly sensitive and reliable quantitative detection of adenosine triphosphate (ATP) in biological fluids. The platform utilizes gold-Prussian blue core-shell nanoparticles modified with polyethyleneimine (Au@PB@PEI NPs), embedded within gold nanoparticle-immobilized capillary-based silica monolithic materials. PB acts as an internal standard, while PEI enhances molecular capture. The periodic, bimodal porous structure of the silica monolithic materials provides uniform and abundant sites for nanoparticle attachment, facilitating rapid liquid permeation, intense SERS enhancement, and efficient enrichment. The platform regulates ATP capture and release through magnesium ions in the liquid phase, eliminating matrix interferences and enabling platform reuse. Integrating efficient molecular enrichment, separation, an interference-free internal standard, a liquid flow channel, and a detection chamber, our platform offers simplicity in operation, exceptional sensitivity and accuracy, and rapid analysis (∼10 min). Employing PB as an internal calibration standard, ratiometric Raman signals (I732/I2123) facilitate precise ATP quantification, achieving a remarkable limit of detection down to 0.62 pM. Furthermore, this platform has been proven to be highly reproducible and validated for ATP quantification in both mouse cerebrospinal fluid and human serum, underscoring its immense potential for POCT applications.

Metabolic signatures of metabolites of the purine degradation pathway in human plasma using HILIC UHPLC–HRMS

The metabolic disorders in the purine degradation pathway have proven to be closely associated with several human diseases. However, the etiology is not yet fully understood. Profile assay of purine intermediates and uric acid involved in the metabolic pathway can provide additional insight into the nature and severity of related diseases. Purine metabolites are endogenous chemicals with high hydrophilicity, polarity, and similar structures, thus there is a great need for a specific method to quantify them directly in biological fluids with a short running time. Herein, eight purine degradation pathway metabolites, including xanthine, hypoxanthine, guanine, xanthosine, inosine, guanosine, adenosine and uric acid, in human plasma were quantitatively measured using hydrophilic interaction chromatography-tandem high-resolution mass spectrometry (HILIC-HRMS) in a short running time of 10 min. The method was systematically validated for specificity, linearity of the calibration curve, the limit of detection, the limit of quantification, the lower limit of quantification, precision, accuracy, extraction recovery, matrix effect, and stability. The results showed that the method was linear (R2 > 0.99), accurate (the intra- and inter-day recoveries of all analytes ranged from 90.0 % to 110.0 %), and precise (the intra- and inter-day precisions were less than 6.7 % and 8.9 %, respectively) with the lower limits of quantification ranging from 3 to 10,000 ng/mL. The extraction recoveries and matrix effects were repeatable and stable. All the analytes were stable in the autosampler and could be subject to three freeze-thaw cycles. The developed method was ultimately applied to 100 plasma specimens from healthy individuals. The results showed that the concentrations of different purine metabolites varied dramatically in plasma specimens. Diet and body mass index (BMI) were the most significant factors determining purine levels, followed by drinking and sex. Age, smoking and bedtime showed a very weak correlation with purine metabolism. The findings of the present work reveal the characteristics of purine metabolism in human plasma under non-pathological conditions. The results also highlight the factors that can cause changes in purine metabolism, which are useful in developing effective treatment strategies for metabolic disorders of purines, particularly for those caused by lifestyle factors.

Silica particle size and polydispersity control from fundamental practical aspects of the Stöber method

Silica-based nanoparticles have been continuously investigated for their physical and chemical properties, which have made them adaptable for a wide range of scientific purposes, from adsorption in an aqueous medium to target-specific drug delivery in biological fluids. The achievement of these properties is, in general, possible by controlling the synthesis of the nanomaterial, and the Stöber method is one of the most commonly employed approaches for silica-based nanoparticles. In this study, experiments were carried out to investigate several parameters of the Stöber method for controlling the diameter and size dispersion of silica nanoparticles. The results of these experiments were supported by and contrasted with the theoretical aspects of silica particle nucleation and growth models. In this scenery, practical aspects of the Stöber Method such as the effects of variations in catalyst concentration, colloidal stability and drying approach on particle size and polydispersity were examined in detail for providing an organized basis of experimental and theoretical data about it for who aims at work with silica nanoparticle synthesis through the Stöber Method.

A method development for sertraline detection in the blood

Aim – to develop a method for isolation and determination of sertraline in biological fluids. Material and methods. For the study we used "Zoloft" drug ("Pfizer", USA). The study equipment and substances: liquid chromatograph Shimadzu LC-20 Prominence (Japan) with detector SPD-M20A, gas chromatograph Agilent Technologies (USA) 7890 A / 5977 MSD, (program MassHunter GC-MS ), waters vacuum unit and cartridges for solid-phase extraction Oasis HLB; papain enzymes (MedFloran, Russia); trypsin, chymotrypsin, hyaluronidase (Lidaza) (Samson-Med LLC, Russia); test strips from different manufacturers under trade names: "Be sure", "NarcoCHEC", "FACTOR-MED". Urine samples were obtained after administration of the drug to laboratory animals – guinea pigs. The base of sertraline from a solution of its salt was isolated by liquid-liquid extraction (LLE) with organic solvents and their mixtures at pH = 10, 11 and 12 (pKa sertraline 9.48). Obtaining data on the effectiveness of blood sample preparation methods (liquid-liquid extraction, solid-phase extraction and enzymatic hydrolysis) for sertraline was carried out on enriched model samples of donor blood according to the method proposed by Cheger. Results. Cross-positive urine test results were obtained with immunochromatographic test strips for derivatives of e 1,4-benzodiazepine and synthetic cannabimimetics ("spice"). The study with 1 mg/ml solution of sertraline gave a negative result. Desmethylsertraline, a major metabolite, was detected in the urine, a peak with a retention time of about 12.75 min, the peak of the original substance during the synthesis of sertraline – sertralinimine and the peak of the native sertraline with a retention time of about 12.65 min. A method for detecting sertraline by HPLC has been developed. Conclusion. The use of the enzymatic blood hydrolysis with trypsin allows for increasing the degree of extraction by 1.5 times when compared with liquid-liquid extraction with chloroform, and by 2 times when compared with solid-phase extraction.

Diaper-based wearable SERS sensing system with a silver nano dual-structure composite hydrogel for the detection of biomarkers and pH in urine

Wearable sensors are essential for continuous monitoring of an individual’s health status to enable personalized health management. Urine as a typical non-invasive biological fluid is rich in biological information. Although diaper-based biosensors have been developed, the development of wearable sensors for multiple biomarkers in urine remains a major challenge to be explored. In this paper, a hydrogel SERS sensor consisting of two metal nanostructures of silver nanowires (Ag NWs) and silver nanocubes (Ag NCs) was developed and adhered to nappies to form a wearable nappy-based SERS sensor for the rapid and sensitive detection of bio markers (creatinine, uric acid and bilirubin) and pH in urine. The hydrogel’s excellent adsorption and water-locking properties greatly simplify the collection of urine samples. At the same time, its encapsulation effect can effectively maintain the freshness of the urine sample and ensure the accuracy of the test. The introduction of silver nano dual-structure improves the antimicrobial properties of the hydrogel and reduces the risk of infection when the sensor comes into contact with the skin. Moreover, the silver nano dual-structure triggers abundant electromagnetic “hot spots” around intersecting nanojunctions and gaps, which greatly improves SERS sensitivity. For the detection of creatinine, uric acid and bilirubin, the LODs were as low as 0.59 μM, 69 nM, 89 nM. Finally, the urine pH was accurately detected using a functionalized Ag NWs-Ag NCs/CCNF/Gel/PVA hydrogel. This wearable diaper sensor is expected to be used for personal health condition monitoring and holds great promise for effective care of infants, the elderly, and those with limited mobility.

Studies of the interaction of graphene oxide (GO) with endothelial cells under static and flow conditions

Graphene oxide, due to its unique properties, has several potential applications in biomedicine, especially as a drug carrier. Despite emerging studies on its cytotoxicity and uptake into cells, there are still gaps in knowledge on this area. When analyzing the internalization of nanomaterials, many different factors must be considered, including particle size, surface modifications, and interactions with biological fluids that can change their properties. In the present study, we evaluated the effects of graphene oxide fractions in different sizes and samples incubated in human serum on endothelial cells (HUVECs). In addition, the study was conducted in both macroscale and microscale using Cell-on-a-Chip technology to better replicate in vivo conditions. Our findings indicate that samples incubated with serum reduce the efficiency of fraction uptake into cells. It was also observed that the uptake efficiency of graphene oxide (GO) fractions is higher in the microscale (in more real to in vivo environment) compared to the macroscale. Our research has shown that in order to determine the correct interaction of new materials into mammalian cells, it is necessary to take into account many different biochemical and physical factors.

Mathematical analysis of isothermal study of reverse roll coating using Micropolar fluid

This article demonstrates a mathematical model and theoretical analysis of the Micropolar fluid in the reverse roll coating process. It is important because micropolar fluids account for the microstructure and microrotation of particles within the fluid. These characteristics are significant for accurately describing the behavior of complex fluids such as polymer solutions, biological fluids, and colloidal suspensions. First, we modeled the flow equations using basic laws of fluid dynamics. The flow equations are made modified using low Reynolds number theory. The simplified equations are solved analytically. The exact expression for velocity and pressure gradient are obtained, while pressure is calculated numerically using Simpson Rule. Graphical depictions are carried out to comprehend the impact of the newly emerged physical constraints. The influence of micropolar and microrotation parameters on the velocity, pressure and pressure gradient are elaborated with the help of different graphs.

SPRi Biosensor for Simultaneous Determination of HIF-1α, Angiopoietin-2, and Interleukin-1β in Blood Plasma.

A new analytical method, based on SPRi biosensors, has been developed for the simultaneous determination of the pro-angiogenic factors HIF-1α, angiopoietin-2 (ANG-2), and interleukin-1β (IL-1β) in biological fluids. These proteins take part in the process of angiogenesis, i.e., the creation of new blood vessels, which is a key stage of cancer development and metastasis. A separate validation process was carried out for each individual compound, indicating that the method can also be used to study one selected protein. Low values of the limit of detection (LOD) and quantification (LOQ) indicate that the developed method enables the determination of very low concentrations, in the order of pg/mL. The LOD values obtained for HIF-1α, ANG-2, and IL-1β were 0.09, 0.01, and 0.01 pg/mL, respectively. The LOQ values were 0.27, 0.039, and 0.02 pg/mL, and the response ranges of the biosensor were 5.00-100.00, 1.00-20.00, and 1.00-15.00 pg/mL. Moreover, determining the appropriate validation parameters confirmed that the design offers high precision, accuracy, and sensitivity. To prove the usefulness of the biosensor in practice, determinations were made in plasma samples from a control group and from a study group consisting of patients with diagnosed bladder cancer. The preliminary results obtained indicate that this biosensor can be used for broader analyses of bladder cancer. Each of the potential biomarkers, HIF-1α, ANG-2, and IL-1β, produced higher concentrations in the study group than in the control group. These are preliminary studies that serve to develop hypotheses, and their confirmation requires the analysis of a larger number of samples. However, the constructed biosensor is characterized by its ease and speed of measurement, and the method does not require special preparation of samples. SPRi biosensors can be used as a sensitive and highly selective method for determining potential blood biomarkers, which in the future may become part of the routine diagnosis of cancers.

The application of vibrational spectroscopy in forensic analysis of biological evidence.

Vibrational spectroscopy is a powerful analytical domain, within which Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy stand as exemplars, offering high chemical specificity and sensitivity. These methodologies have been instrumental in the characterization of chemical compounds for an extensive period. They are particularly adept at the identification and analysis of minute sample quantities. Both FTIR and Raman spectroscopy are proficient in elucidating small liquid samples and detecting nuanced molecular alterations. The application of chemometrics further augments their analytical prowess. Currently, these techniques are in the research phase within forensic medicine and have yet to be broadly implemented in examination and identification processes. Nonetheless, studies have indicated that a combined classification model utilizing FTIR and Raman spectroscopy yields exceptional results for the identification of biological fluid-related information and the determination of causes of death. The objective of this review is to delineate the current research trajectory and potential applications of these two vibrational spectroscopic techniques in the detection of body fluids and the ascertainment of causes of death within the context of forensic medicine.

Analytical methods for the determination of xylazine in pharmaceutical, clinical and forensic matrices - A review.

Xylazine, a non-opioid veterinary anaesthetic tranquillizer that is not licensed for human use, has been linked to an increase in overdose fatalities worldwide. The study delves into the forensic aspects of xylazine usage, emphasizing on chemical, clinical and toxicological analyses of drug seizures, bodily fluids and tissues. It advocates for validated analytical methods for determining xylazine. This study provides supporting material to pave the path for the usage and development of relevant and verified alternative screening and confirmation methods for laboratories. Google Scholar, Scopus, Science Direct and PubMed were searched for relevant articles and case reports in relation to xylazine misuse and established analytical methods for forensic investigation until April 2023. A total of 79 articles were evaluated, and 40 publications met the inclusion criteria. The most prevalent xylazine exposures recorded were incidental and intentional misuse/abuse. Common symptoms upon presentation were hypotension, bradycardia, drowsiness and lethargy, although mortality was less prevalent. Solid-phase extraction and liquid-liquid extraction are two extensively used sample preparation techniques. These techniques are used to extract desired analytes from complex matrices. Several analytical techniques have been stated, including GC-MS, LC-MS/MS, HPLC-DAD and others. The analytical procedures used are determined by the matrices involved, the amount of xylazine present, interfering compounds, the degree of precision required and the laboratory infrastructure. In the present context, the LC-MS/MS methods are preferred as the gold standard. In the near future, many analytical techniques such as capillary electrophoresis, PSI-MS, immuno-analytical techniques and SERRS may show significant potential.

Knowledge of Hepatitis-B Management and its Prevention among the Senior Staff Nurses Working at Bangabandhu Sheikh Mujib Medical University (BSMMU)

Background: Hepatitis B, caused by the Hepatitis B virus (HBV), presents a significant global health challenge, especially in regions like Bangladesh where its prevalence is intermediate. The virus is transmitted through percutaneous or mucosal exposure to infectious blood or bodily fluids, posing a substantial risk to healthcare workers. Hepatitis B can lead to severe liver diseases if untreated. In Bangladesh, the burden of Hepatitis B is high, necessitating urgent intervention, particularly among healthcare professionals such as senior staff nurses who are on the frontline of managing and preventing the disease. Methods: This cross-sectional study assessed the knowledge levels of 210 senior staff nurses at Bangabandhu Sheikh Mujib Medical University (BSMMU) regarding Hepatitis B management and prevention. A structured questionnaire, translated into Bengali, was used for data collection through structured interviews. Convenience sampling was employed, and data were analyzed using SPSS (Version 24.0). Descriptive statistics and t-tests/ANOVA were used to explore the relationship between socio-demographic characteristics and nurses' knowledge about Hepatitis B. Ethical clearance was obtained from the Institutional Review Board (IRB) of BSMMU. Results: The majority of respondents were female (70%), aged below 30 years (59.5%), and had basic nursing education (89.5%). Most had 1-10 years of work experience (89%) and earned less than 35,000 BDT monthly (78.1%). Regarding vaccination, 64.3% had completed it, 26.7% were partially vaccinated, and 9% were unvaccinated. Occupational exposure to blood or body fluids was reported by 73.3% of nurses. Knowledge about Hepatitis B infection, transmission, and prevention was generally high, with 85.44%, 89.14%, and 85.11% of nurses demonstrating good knowledge in these areas, respectively. Marital status and vaccination status were significantly associated with higher knowledge levels (p < 0.05). Conclusion: Senior staff nurses at BSMMU exhibit substantial knowledge about Hepatitis B infection, transmission, and prevention. However, misconceptions persist regarding non-transmission routes and some preventive measures. Married nurses and those with complete vaccination status demonstrated higher knowledge levels. These findings highlight the need for targeted educational interventions to address knowledge gaps and reinforce preventive practices among nurses. This study underscores the importance of continuous education and vaccination programs to enhance the effectiveness of Hepatitis B management and prevention efforts in healthcare settings.

Potential of high-resolution mass spectrometry for identification and structural elucidation of scopolamine metabolomic biomarkers in a confirmed case of Brugmansia intoxication. Specially application in drug-facilitated crimes

In forensic toxicology, scopolamine remains as one of the most challenging alkaloid in terms of analytical detection. Given its rapid elimination, the detection window in common matrices is short. Taking advantage of a real case of Brugmansia intoxication, a metabolic study was carried out. We report the real case of a 16-year-old boy admitted to an Emergency Unit after consumption at high school of a beverage made of Brugmansia dried flowers. The medical staff noticed agitation, mydriasis and tachycardia. Scopolamine and atropine were positively detected in biological fluids using liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS/MS). To better characterize the intake and identify metabolic biomarkers, we developed a data mining workflow specific to tropane alkaloids and applied it to the urine sample. This metabolic profile may be useful in providing analytical methods with a wider detection window particularly in drug-facilitated crimes (DFC). Scopolamine and atropine metabolites were predicted in silico with GLORYx freeware to assist in metabolite identification. The previously published metabolic pathways for scopolamine and atropine in mammals were studied as well. A total of fifteen phase I and II metabolites were tentatively identified for scopolamine, while one metabolite was detected for atropine. In addition, we identified some tropane alkaloids from the plant that were also metabolized. These metabolites can be used as biomarkers of exposure to Solanaceae plants and may also be useful to distinguish between natural product use and clinical therapy.

Self-Assembled Monolayer Transporters Enable Reagentless Analysis of Small Molecule Analytes.

The detection of small molecules beyond glucose remains an ongoing challenge in the field of biomolecular sensing owing to their small size, diverse structures, and lack of alternative non-enzymatic sensing methods. Here, we present a new reagentless electrochemical approach for small molecule detection that involves directed movement of electroactive analytes through a self-assembled monolayer to an electrode surface. Using this method, we demonstrate detection of several physiologically relevant small molecules as well as the capacity for the system to operate in several biological fluids. We anticipate that this mechanism will further improve our capacity for small molecule measurement and provide a new generalizable monolayer-based technique for electrochemical assessment of various electroactive analytes.

Development of borazine-assisted-oriented molecularly imprinted electrochemical sensor for the detection of umifenovir in serum and urine by EIS and DPV methods

Herein, borazine (B3N3) assisted-oriented surface imprinted electrochemical sensor was designed for umifenovir (UMI) detection in serum and urine samples. An imprinted electrochemical sensor was developed via photopolymerization whereas butyl methacrylate monomer (BuMA) was selected as the functional monomer for coordinating the template molecules, UMI in the presence of B3N3 as orientating UMI molecules between polymeric phase and the electrode surface. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to verify the electrochemical characterizations of the alterations at each step of the MIP production process. The created sensor was also characterized via contact angle, Fourier transform infrared spectrophotometry attenuated total reflection (FTIR-ATR), atomic force spectroscopy (AFM), and scanning electron microscopy (SEM) measurements. Furthermore, the electronic and molecular alterations on the electrode surface were assessed using quantum chemical calculations. Using differential pulse voltammetry (DPV) and EIS methods in standard solution and biological fluid samples, UMI was determined separately. The dynamic linear range of the designed sensor under optimized experimental conditions was found to be 0.50–7.50 pM and 0.25–5.00 pM for DPV and EIS methods, respectively. The developed sensor exhibited good recognition ability towards UMI molecules in the presence of its analogues as well as other interferents. As a result, the proposed sensors demonstrated excellent selectivity, high sensitivity, repeatability, and reproducibility performance for UMI analysis in commercial serum and urine samples.

Forensic efficiency evaluation of a mtDNA whole genome sequencing system constructed with long fragment amplification strategy on DNA nanoball sequencing platform

Mitochondrial DNA (mtDNA) is an important genetic marker for degraded biological sample identification, maternal pedigree tracing, and population genetic structure study owing to its characteristics of high copy number, anti-degradable ring structure, and maternal inheritance. Whole mtDNA genome sequencing is an optimal method for the analysis of mtDNA polymorphism and heterogeneity because it allows for the comprehensive use of maternal genetic information. However, because of lacking quantitative evaluations for sequencing data, the scientific interpretation standards for mtDNA sequencing results of the previously used sequencing systems are often different, and false positive or false negative results are prone to occur when faced with the interference of nuclear genomic DNA, or the heterogeneities of mtDNA sequence and structure. In this study, we evaluated a novel mtDNA whole genome sequencing system using long fragment amplification strategy on the DNA nanoball (DNB) sequencing platform. This system demonstrated high sequencing quality and specific mtDNA sequencing efficiencies on positive control DNA and FTA bloodstain samples, as the average Q20 and Q30 values of the corresponding samples were 97.17 % and 91.93 %; 97.37 % and 92.48 %, respectively. The mean mapping percentages for the reference sequences of whole genome DNA (wgDNA), mtDNA, and nuclear genomic DNA (ngDNA) in the corresponding samples were 99.98 %, 99.97 %, 0.03 %, and 99.91 %, 99.40 %, 0.60 %; respectively. The average error calling rates for the bases A, C, G, and T of the whole mtDNA genome were 0.2519 %, 0.2550 %, 0.2906 %; and 0.2392 %, respectively. The efficacy of heteroplasmy identification was assessed using a set of theoretical sites with predetermined rates. These sites were created by combining the samples with known mtDNA haplotypes in certain proportions. The absolute errors between observed and theoretical heteroplasmy values were 89.59 %, 74.68 %, 50.20 %, 12.65 %, 8.31 %, and 4.85 %, while the theoretical heteroplasmy values were 5 %, 10 %, 20 %, 80 %, 90 %, and 95 %, respectively. The absolute error exhibited relative stability when the mtDNA sequencing depth exceeded 500×. Furthermore, the system sequencing efficiency was also confirmed among different kinds of samples, and these samples included natural samples (e.g., peripheral blood samples preserved on FTA cards for 2 and 11 years, and on filter paper for 6 and 9 years), degraded samples, sensitivity samples, samples derived from various bodily fluids, and maternal pedigree samples. In summary, the whole mtDNA genome sequencing system used for forensic identification demonstrated high performance in analyzing mtDNA sequence information, and showed significant prospects for forensic application and maternal genetic research.