Non-invasive Biological Samples Research Articles

Semiconductor-Based Biosensor Functionalized with Jacalin for Detection of Secretory IgA in Sweat Sample

Recently, mental stress-related diseases, such as integration disorder syndrome and depression, affect people's health. Previously, the relation between concentration of stress-markers and mental stress condition has been reported, suggesting that a simple and daily detection of these markers from biological samples is important to prevent the disease. Secretory immunoglobulin A (s-IgA) which is present in the non-invasive biological samples, such as sweat or saliva, is one of the candidates to measure the mental stress. For the detection of s-IgA, field effect transistor (FET) biosensor is a promising device to easily and quickly detect biomolecules, as the FET characteristics can be responded to the change of surface potential caused by the adsorption of charged molecules. To achieve a sensitive detection by using FET biosensors, small receptors have been applied for an effective use of charge-recognition region, Debye length [1]. In this study, jacalin, a small tetrameric lectin from plant (66 kDa), was selected as a receptor to detect s-IgA. Each monomer of jacalin has a glycan-binding site to the hinge region of IgA, thus the sensitivity expected to be improved due to increasing the amounts of specifically adsorbed s-IgA. The SiO2 gate surface was exposed to O2 plasma (200 W for 1 min) in order to introduce hydroxyl groups reacting with triethoxysilane groups of self-assembled monolayer (SAM). Then, FET was immersed in 1%(v/w) 3-aminopropyltriethoxysilane toluene solution at 60ºC for 7 min in an argon atmosphere, followed by the cross-linking by glutaraldehyde. After the modification with the cross-linker, jacalin was immobilized on FET gate surfaces, and ethanolamine capping was performed to prevent the non-specific adsorption of contaminating molecules. The electrical responses were calculated by the FET characteristics before and after the addition of target solution. The FET characteristics were measured by sweeping the gate-voltage from -2.0 V to 0 V with 0.1 V drain voltage in 0.01 × phosphate buffered saline (pH 7.4). The immobilization of jacalin through the aminopropylsilane SAM was confirmed from atomic force microscopic images on the surface morphology change of the FET gate surfaces. Additionally, to optimize the immobilization condition, the relationship between concentration of jacalin solution and magnitudes of electrical responses were examined. Then, the specificity of jacalin-immobilized FET biosensor to s-IgA were confirmed by the comparison of responses caused by the addition of s-IgA and human serum albumin (HSA). The ΔV g was shifted in a positive direction due to the negative charges of s-IgA (isoelectric point = 4.5-5.0). On the other hand, when the HSA was added to the FET biosensor, the responses was scarcely observed, suggesting that the jacalin-immobilized surface specifically captured the s-IgA (Figure 1). In addition, a correlation between the FET responses and s-IgA from 100 ng/mL to 100 μg/mL, including the fluctuations of concentration in human sweat sample, was clearly obtained. These results suggested that jacalin could be useful as a receptor for FET biosensors to achieve a simple and sensitive detection of s-IgA. [1] S. Cheng, K. Hotani, S. Hideshima, S. Kuroiwa, T. Nakanishi, M. Hashimoto, Y. Mori and T. Osaka, Materials, 7, (4), 2490-2500, (2014). Figure 1

Recruitment and Retention of Pregnant and Breastfeeding Mothers for Longitudinal Clinical Microbiome Studies (OR30-08-19)

ObjectivesAccumulating data suggest the health benefits of breastfeeding are, in part, mediated through interactions with the human microbiome. Despite these observations, recruitment of pregnant mothers in longitudinal clinical microbiome studies remains a significant challenge. The goal of this study is to develop data-driven strategies for recruitment and retention of breastfeeding mothers’ into longitudinal clinical microbiome studies. MethodsWe recruited a total of 40 mothers (20 pregnant and 20 breastfeeding) to complete in-depth semi-structured interviews at the UF Clinical and Translational Science Institute. Additionally, mothers were asked to use self-administered microbiome collection kits and complete questionnaires related to health history, infant feeding practices, and physical activity. Informative interviews were designed to identify women’s understanding, motivations and barriers surrounding longitudinal clinical research participation and their preferences for providing non-invasive biological samples. ResultsOur preliminary analysis indicates that 1) barriersfor participating in research include convenience, confidentiality and child protection; 2) factors motivating participation in research include general interest in breastfeeding, schedule, and perceived research relevance; 3) participant recruitment should consider social media and 4) biological sample collection should include adequate instructions and drop-off convenience. ConclusionsThe results of this study provide valuable theoretical and practical insights on how to effectively recruit and retain pregnant and breastfeeding women into longitudinal microbiome studies. Funding Sources1K01DK115632-01A1.

Validated analytical method to determine new salivary lipid peroxidation compounds as potential neurodegenerative biomarkers

Lipid peroxidation is closely related to neurodegenerative diseases since brain shows high lipid composition and oxygen consumption. The determination of lipid peroxidation compounds in non-invasive biological samples would help to monitor the patients’ oxidative stress status. A new analytical method based on ultrasound-assisted liquid-liquid semi-microextraction (UA-LLsME) followed by Ultra Performance Liquid Chromatography coupled to tandem Mass Spectrometry was developed to determine 18 lipid peroxidation biomarkers in saliva samples. The variables affecting the UA-LLsME efficiency were systematically studied. Under the optimum conditions, the methodology was validated and showed high-throughput, high sensitivity (limits of detection 0.02-2 nmol L−1), and satisfactory precision (coefficients of variation 2–11% (intra-day) and 5–12% (inter-day)). The reliability of the described method was assessed analysing spiked saliva samples, and the recoveries were between 80% and 120% for most of the analytes. Then, the method suitability was demonstrated by analysing saliva samples (n = 30) from elderly people with neurodegenerative diseases. To conclude, the new developed analytical method is a useful tool to determine salivary lipid peroxidation compounds as potential biomarkers in further clinical studies in which oxidative stress plays an important role.

Manganese in toenails is associated with hearing loss at high frequencies in humans

Purpose: Elevated hearing thresholds from high frequencies are known to be one of the hallmarks of age-related hearing loss. Our recent study showed accumulation of manganese (Mn) in inner ears resulting in acceleration of age-related hearing loss in mice orally exposed to Mn. However, there is no evidence showing an association between Mn in non-invasive biological samples and hearing loss in humans evaluated by pure tone audiometry (PTA). In this study, we evaluated Mn in non-invasive biological samples as a possible biomarker for hearing loss in humans.Materials and methods: We determined hearing levels by PTA and Mn levels in toenails, hair and urine with an inductively coupled plasma mass spectrometer (ICP-MS) in 145 healthy subjects in Bangladesh.Results: Multivariable analyses showed that Mn levels in toenails, but not in hair and urine samples, were significantly associated with hearing loss at 8 kHz and 12 kHz. Moreover, our experimental study showed a significant correlation between Mn levels in inner ears and nails, but not hair, in mice orally exposed to Mn.Conclusions: The results provide novel evidence that Mn in toenails is a possible biomarker for hearing loss at high frequencies in humans.

Methadone Concentrations in Exhaled Breath Condensate, Serum and Urine of Patients Under Maintenance Treatment.

Drug abuse is a serious problem causing health, economical and psycho-social negative outcomes. Methadone is commonly used drug for management of drug addiction. Exhaled breath condensate (EBC) is a promising non-invasive biological sample which attracted more attention in recent years. This work aimed to extend the applicability of a developed preconcentration - liquid chromatographic method for analysis of methadone in serum and urine samples. Drug concentrations in EBC, serum and urine are also investigated for dose-concentration and their inter-correlations. Biological samples were collected from 53 patients receiving methadone and the concentrations were determined using a validated analytical method after a pre-concentration step. Methadone measured in all samples and there are correlations between administered dose of methadone and its serum and urine concentrations. A weak correlation is observed between dose and EBC concentration. Wider variations in EBC concentrations of methadone could be justified concerning a number of affecting parameters such as relative humidity of the collection area and further investigations are required for standardization of EBC as a non-invasive biological sample to be used in clinical practice.

Analysis of deferiprone in exhaled breath condensate using silver nanoparticle-enhanced terbium fluorescence

Exhaled breath condensate (EBC) has been proposed as an alternative non-invasive biological sample for therapeutic drug monitoring.

Urinary micro-RNA biomarker detection using capped gold nanoslit SPR in a microfluidic chip.

Successful diagnosis and treatment of many diseases depends on the availability of sensitive, reliable and low cost tools for the detection of the biomarkers associated with the diseases. Simple methods that use non-invasive biological samples are especially suitable for the deployment in the clinical environment. In this paper we demonstrate the application of a method that employs a capped gold nanoslit surface plasmon resonance (SPR) sensor and a microfluidic chip for the detection of a urinary nucleic acid biomarker in clinical samples. This method detects low concentrations of the biomarker in a relatively large volume (∼1 mL) of the sample. The method utilizes magnetic nanoparticles (MNPs) for the isolation of target molecules and signal enhancement in conjunction with surface plasmon resonance (SPR) on capped gold nanoslits. The ability of the method to detect urinary miRNA-16-5p in AKI patients was tested and the result was compared with the data obtained with the polymerase chain reaction (PCR). miRNA-16-5p has been found to be a specific and noninvasive biomarker for acute kidney injury (AKI). Our method allows the detection of the biomarker in the urine of AKI patients without amplification and labeling of the target molecules.

Correction: Evaluation of Non-Invasive Biological Samples to Monitor Staphylococcus aureus Colonization in Great Apes and Lemurs

Correction: Evaluation of Non-Invasive Biological Samples to Monitor Staphylococcus aureus Colonization in Great Apes and Lemurs

Evaluation of Non-Invasive Biological Samples to Monitor Staphylococcus aureus Colonization in Great Apes and Lemurs

IntroductionReintroduction of endangered animals as part of conservational programs bears the risk of importing human pathogens from the sanctuary to the natural habitat. One bacterial pathogen that serves as a model organism to analyze this transmission is Staphylococcus aureus as it can colonize and infect both humans and animals. The aim of this study was to evaluate the utility of various biological samples to monitor S. aureus colonization in great apes and lemurs.MethodsMucosal swabs from wild lemurs (n=25, Kirindy, Madagascar), feces, oral and genital swabs from captive chimpanzees (n=58, Ngamba and Entebbe, Uganda) and fruit wadges and feces from wild chimpanzees (n=21, Taï National Parc, Côte d’Ivoire) were screened for S. aureus. Antimicrobial resistance and selected virulence factors were tested for each isolate. Sequence based genotyping (spa typing, multilocus sequence typing) was applied to assess the population structure of S. aureus.ResultsOro-pharyngeal carriage of S. aureus was high in lemurs (72%, n=18) and captive chimpanzees (69.2%, n=27 and 100%, n=6, respectively). Wild chimpanzees shed S. aureus through feces (43.8, n=7) and fruit wadges (54.5, n=12). Analysis of multiple sampling revealed that two samples are sufficient to detect those animals which shed S. aureus through feces or fruit wadges. Genotyping showed that captive animals are more frequently colonized with human-associated S. aureus lineages.ConclusionOro-pharyngeal swabs are useful to screen for S. aureus colonization in apes and lemurs before reintroduction. Duplicates of stool and fruit wadges reliably detect S. aureus shedding in wild chimpanzees. We propose to apply these sampling strategies in future reintroduction programs to screen for S. aureus colonization. They may also be useful to monitor S. aureus in wild populations.

Non-invasive reproductive and stress endocrinology in amphibian conservation physiology.

Non-invasive endocrinology utilizes non-invasive biological samples (such as faeces, urine, hair, aquatic media, and saliva) for the quantification of hormones in wildlife. Urinary-based enzyme immunoassay (EIA) and radio-immunoassay have enabled the rapid quantification of reproductive and stress hormones in amphibians (Anura: Amphibia). With minimal disturbance, these methods can be used to assess the ovarian and testicular endocrine functions as well as physiological stress in captive and free-living populations. Non-invasive endocrine monitoring has therefore greatly advanced our knowledge of the functioning of the stress endocrine system (the hypothalamo-pituitary-interrenal axis) and the reproductive endocrine system (the hypothalamo-pituitary-gonadal axis) in the amphibian physiological stress response, reproductive ecology, health and welfare, and survival. Biological (physiological) validation is necessary for obtaining the excretory lag time of hormone metabolites. Urinary-based EIA for the major reproductive hormones, estradiol and progesterone in females and testosterone in males, can be used to track the reproductive hormone profiles in relationship to reproductive behaviour and environmental data in free-living anurans. Urinary-based corticosterone metabolite EIA can be used to assess the sublethal impacts of biological stressors (such as invasive species and pathogenic diseases) as well as anthropogenic induced environmental stressors (e.g. extreme temperatures) on free-living populations. Non-invasive endocrine methods can also assist in the diagnosis of success or failure of captive breeding programmes by measuring the longitudinal patterns of changes in reproductive hormones and corticosterone within captive anurans and comparing the endocrine profiles with health records and reproductive behaviour. This review paper focuses on the reproductive and the stress endocrinology of anurans and demonstrates the uses of non-invasive endocrinology for advancing amphibian conservation physiology. It also provides key technical considerations for future research that will increase the accuracy and reliability of the data and the value of non-invasive endocrinology within the conceptual framework of conservation physiology.

Endometrial fluid is a specific and non-invasive biological sample for protein biomarker identification in endometriosis

Endometrial fluid is a specific and non-invasive biological sample for protein biomarker identification in endometriosis

Endometrial fluid is a specific and non-invasive biological sample for protein biomarker identification in endometriosis

The development of non-invasive diagnostic methods for endometriosis requires sensitive and disease specific biomarkers. Here, we describe the use of aspirated endometrial fluid from women with and without endometriosis as a novel biological sample for biomarker discovery. Differential protein expression profiling of aspirates from women with early endometriosis (n = 14), advanced endometriosis (n = 32) and without evidence of the disease (n = 32) was assessed by two-dimensional gel electrophoresis (2-DE). A biomarker validation study was performed in an independent cohort (early endometriosis n = 6 and advanced endometriosis n = 14, controls n = 15). The analysis resulted in the identification of 31 proteins showing statistically significant differences in expression. The proteins identified are related to cell signalling, cell death and cell movement, processes that may be involved in the onset and/or progression of endometriosis. The differences in expression observed for 14-3-3 (signal transduction) and moesin (cytoskeletal structure) were confirmed in an independent group of endometriosis patients. Endometrial fluid represents a novel sample for proteomic analysis offering reliable, disease specific information on protein expression, facilitating the discovery of biomarkers for endometriosis. The results described here complement previous proteomic studies, providing new endometriosis-related proteins to be validated as diagnostic markers.

Feasibility of collecting oral fluid samples in the home setting to determine seroprevalence of infections in a large-scale cohort of preschool-aged children

Oral fluid is a non-invasive biological sample, which can be returned by post, making it suitable for large-scale epidemiological studies in children. We report our experience of oral fluid collection from 14 373 preschool-aged children in the UK Millennium Cohort Study. Samples were collected by mothers in the home setting following the guidance of trained interviewers, and posted to the laboratory. Samples were received from 11 698 children (81.4%). Children whose mothers were of Black Caribbean ethnicity and who lived in non-English-speaking households were less likely to provide a sample, and those with a maternal history of asthma more likely to provide a sample [adjusted risk ratio (95% CI) 0.85 (0.73-0.98), 0.87 (0.77-0.98) and 1.03 (1.00-1.05) respectively]. Collection of oral fluid samples is feasible and acceptable in large-scale child cohort studies. Formal interpreter support may be required to increase participation rates in surveys that collect biological samples from ethnic minorities.