Polycystic ovary syndrome: clinical and laboratory variables related to new phenotypes using machine-learning models.

PURPOSE: Polycystic Ovary Syndrome (PCOS) is the most frequent endocrinopathy in women of reproductive age. Machine learning (ML) is the area of artificial intelligence with a focus on predictive computing algorithms. We aimed to define the most relevant clinical and laboratory variables related to PCOS diagnosis, and to stratify patients into different phenotypic groups (clusters) using ML algorithms. METHODS: Variables from a database comparing 72 patients with PCOS and 73 healthy women were included. The BorutaShap method, followed by the Random Forest algorithm, was applied to prediction and clustering of PCOS. RESULTS: Among the 58 variables investigated, the algorithm selected in decreasing order of importance: lipid accumulation product (LAP); abdominal circumference; thrombin activatable fibrinolysis inhibitor (TAFI) levels; body mass index (BMI); C-reactive protein (CRP), high-density lipoprotein cholesterol (HDL-c), follicle-stimulating hormone (FSH) and insulin levels; HOMA-IR value; age; prolactin, 17-OH progesterone and triglycerides levels; and family history of diabetes mellitus in first-degree relative as the variables associated to PCOS diagnosis. The combined use of these variables by the algorithm showed an accuracy of 86% and area under the ROC curve of 97%. Next, PCOS patients were gathered into two clusters in the first, the patients had higher BMI, abdominal circumference, LAP and HOMA-IR index, as well as CRP and insulin levels compared to the other cluster. CONCLUSION: The developed algorithm could be applied to select more important clinical and biochemical variables related to PCOS and to classify into phenotypically different clusters. These results could guide more personalized and effective approaches to the treatment of PCOS.

Natural Materials Modified and Applied to the Detection of Drugs In Situ: Modification of Eggshell and Quantification of Oxytetracycline.

This work describes a novel sensing system using eggshells as substrate for the first time, targeting the detection and semiquantitative determination of antibiotics in waters from aquaculture, enabling simple, inexpensive, and in situ drug monitoring. Eggshell was ground and the resulting powder was modified by adsorption of suitable reagents, and it takes a typical colour after contact with the antibiotic. The colour intensity is correlated with the concentration of the antibiotic. This novel approach was applied to oxytetracycline, one of the antibiotics commonly used in aquaculture. The chemical changes on the eggshell powder were evaluated and optimised to produce an intense colour change as a function of the concentration of the antibiotic. The colour changes were evaluated by visual comparison with images taken with a digital camera, applying an appropriate mathematical treatment to the colour coordinates of the HSL system used by Windows. The selectivity of the response was tested against other antibiotic drugs. The materials were also used in the analysis of a spiked environmental water sample. Overall, this work presents a rapid, inexpensive, simple and equipment-free method for screening and discrimination of tetracycline drugs in aquaculture. The method is a green approach by reusing eggshells and decreasing the level of contamination correlated to analytical methods, thus being a promising tool for local, rapid, and cost-effective antibiotic monitoring.

Soliton-like structures in the spectrum and the corresponding eigenstates morphology for the quantum desymmetrized Sinai billiard.

By continuously varying certain geometric parameters γ of the totally desymmetrized quantum Sinai billiard, we study the formation of the so-called soliton-like structures in the spectra of the resulting family of systems. We present a detailed characterization of the eigenstate ψn morphologies along such structures. Usually, scarring and bouncing ball mode states are expected to fully explain the solitons. However, we show that they do not exhaust all the possibilities. States with strong resemblance to very particular solutions of the associated integrable case ( 45°- 45° right triangle) also account for the ψn's. We argue that for the emergence of the solitons, in fact, there must be an interplay between the spatial localization properties of the soliton-related ψn's and the rescaling properties of the billiards with γ. This is illustrated, e.g., by comparing the behavior of the eigenwavelengths along the solitons and the billiard size dependence on γ. Considerations on how these findings could extend to other type of billiards are also briefly addressed.

Imprinting Technology in Electrochemical Biomimetic Sensors.

Biosensors are a promising tool offering the possibility of low cost and fast analytical screening in point-of-care diagnostics and for on-site detection in the field. Most biosensors in routine use ensure their selectivity/specificity by including natural receptors as biorecognition element. These materials are however too expensive and hard to obtain for every biochemical molecule of interest in environmental and clinical practice. Molecularly imprinted polymers have emerged through time as an alternative to natural antibodies in biosensors. In theory, these materials are stable and robust, presenting much higher capacity to resist to harsher conditions of pH, temperature, pressure or organic solvents. In addition, these synthetic materials are much cheaper than their natural counterparts while offering equivalent affinity and sensitivity in the molecular recognition of the target analyte. Imprinting technology and biosensors have met quite recently, relying mostly on electrochemical detection and enabling a direct reading of different analytes, while promoting significant advances in various fields of use. Thus, this review encompasses such developments and describes a general overview for building promising biomimetic materials as biorecognition elements in electrochemical sensors. It includes different molecular imprinting strategies such as the choice of polymer material, imprinting methodology and assembly on the transduction platform. Their interface with the most recent nanostructured supports acting as standard conductive materials within electrochemical biomimetic sensors is pointed out.

Screen-printed electrode produced by printed-circuit board technology. Application to Cancer Biomarker Detection by means of plastic antibody as sensing material.

This research work presents, for the first time, a screen-printed electrode (SPE) made on a PCB board with silver tracks (Ag) and a three electrode configuration (AgxO-working, AgxO-counter and Ag/AgxO-reference electrodes), following the same approach as printed-circuit boards (PCBs). This low cost and disposable device was tested for screening a cancer biomarker in point-of-care. The selected biomarker was carcinogenic embryonic antigen (CEA) protein, routinely used to follow-up the progression of specific cancer diseases. The biosensor was constructed by assembling a plastic antibody on the Ag-working electrode area, acting as the biorecognition element of the device. The protein molecules that were entrapped on the polymer and positioned at the outer surface of the polypyrrole (PPy) film were removed by protease action. The imprinting effect was tested by preparing non-imprinted (NPPy) material, including only PPy as biorecognition element. Infrared and Raman studies confirmed the surface modification of these electrodes. The ability of the sensing material to rebind CEA was measured by several electrochemical techniques: cyclic voltammetry (CV), impedance spectroscopy (EIS) and square wave voltammetry (SWV). The linear response ranged from 0.05 to 1.25 pg/mL against logarithm concentration. Overall, producing screen-printed electrodes by means of conventional PCB technology showed promising features, mostly regarding cost and prompt availability. The plastic antibody-based biosensor also seems to be a promising tool for screening CEA in point-of-care, with low response time, low cost, good sensitivity and high stability.

Discrimination of the effects on zebrafish reproduction from pollutants in drinking water via female, via male and/or via fecundation water.

The lack of preventive policy legislation and the low removal rate of organic pollutants in conventional potabilization treatments lead to some of them being present in drinking water. The problem arises because some of these substances have detrimental effects on human reproduction health, via females, via males or even both. In this work, we established the zebrafish as a bioindicator of these types of substances with the goal of discriminating the effects through three different pathways: male, female or water where the fertilization took place. For this purpose, four parameters were analysed: fertility rate, hatching rate and survival and abnormalities rates. So, for each parameter two groups were formed, according to whether adult males or females were reared in bottled spring water (Z) or tap water (B) and if the in vitro fertilization took place in water Z or B. Results revealed a decline in the fertility and hatching rate in water B, due to a water effect. The most plausible explanation could be the presence of substances which affect the micropyle and chorion. Moreover, a decrease in the fertility rate due to an effect over the female was also observed, but in this case by an alteration of the oocyte quality.

Zebrafish as a possible bioindicator of organic pollutants in drinking waters with effects on reproduction: are effects cumulative or reversible?

Organic pollutants are present in drinking waters due to inefficient detection and removal treatments. For this reason, zebrafish is proposed as a complementary indicator in conventional potabilization treatments. Based on the most sensitive parameters detected in our previous work, in this study we attempted to examine the possible cumulative effect between generations of environmental pollutants likely present in drinking waters, when specimens were cultured in the same water and/or the possible reversibility of these effects when cultured in control water. To this end, embryos with the chorion intact were cultured in three drinking waters from different sources and in one control water for up to 5 months in 20 l glass tanks. Four replicates were performed in all water groups. Results in water group C (tap water from a city also located in a region with intensive agricultural activity, but from the hydrological basin of the river Xúquer) revealed a non-reversible effect on fertility rate. Also in water C there was an alteration of sex ratio towards females, although in this case the alteration was reversible. A transgenerational alteration in the germ-line via an epigenetic mechanism from the previous generation is proposed as the most plausible explanation of this effect.

Influence of leaf quality in microbial decomposition in a headwater stream in the Brazilian cerrado: a 1-year study.

The temporal dynamics of leaf decomposition in a tropical stream were evaluated by measuring the input of leaves from riparian vegetation. The resulting mixture of leaves was processed by decomposer microorganisms and, in particular, by aquatic hyphomycetes, the most important microorganisms associated with leaf litter decomposition. Leaf decomposition and the characteristics and activity of the microbial community were investigated over the course of 1 year; these parameters depended on the composition and availability of leaves in the stream. We assessed the biomass, activity, and community structure of the fungi associated with the decomposing leaf mixtures monthly in a tropical stream with little seasonal variation in water characteristics. The leaf material included 27 riparian species; the amount of leaves of each contributing species varied from month to month, with higher overall values in the rainy season. Leaf decomposition was slow, and leaf mass loss did not occur monthly; however, leaf mass loss was significantly lower in May and September. The values of ATP, the ergosterol concentration, and the sporulation of aquatic hyphomycetes varied during the experiment, and only the ergosterol concentration (fungal biomass) was correlated with leaf mass loss. The sporulation rates of aquatic hyphomycetes peaked at the beginning of the rainy season (October), when only three hyphomycete species were present out of the total of seven that were found during the year. In this tropical stream, where the physical and chemical characteristics of the stream water varied little during the year, we hypothesize that changes related to leaf chemical composition could be the cause of changes in decomposition and the abundance and composition of the fungal assemblages. No significant differences were found for the parameters analyzed (lignin, cellulose, fiber, phosphorous, and polyphenols).

Zebrafish (Danio rerio) as a possible bioindicator of epigenetic factors present in drinking water that may affect reproductive function: is chorion an issue?

Emerging organic contaminants have been monitored in stream waters, raw and finished waters and wastewater effluents. Most of these contaminants, such as epigenetic substances, have been detected at very low levels. Unfortunately, their complete monitoring and/or removal are very difficult, given the increasing presence of new contaminants and due to analytical and economic considerations. For this reason, bioindicators are used as an alternative to monitor their presence. To this end, zebrafish is being used to assess certain contaminants in water quality studies. As our long-term aim is to determine if zebrafish (Danio rerio) can be used to detect environmental epigenetic factors in drinking waters with effects on human reproduction, an initial question is whether the chorion could interfere with the possible action of epigenetic factors in two reproductive events: genital ridge formation and migration of the primordial germ cells (PGCs) to these genital ridges. In the first experiment, we attempted to partially degrade the chorion of mid blastula transition (MBT) embryos with pronase, with acceptable survival rates at 5 days post fertilisation (dpf), with the group exposed for 15 min giving the best survival results. As denuded early embryos require a specific culture medium, in the next experiment embryo survival was evaluated when they were cultured up to 5 dpf in drinking waters from six different sources. Results showed a negative effect on embryo survival at 5 dpf from several waters but not in others, thus distorting the survival outcomes. These results suggest using embryos with the chorion intact from the outset when drinking waters from different sources are to be tested.

Zebrafish as a possible bioindicator of organic pollutants with effects on reproduction in drinking waters.

Organic contaminants can be detected at low concentrations in drinking water, raising concerns for human health, particularly in reproduction. In this respect, we attempted to use the zebrafish as a bioindicator to detect the possible presence of these substances in drinking water, aiming to define the most relevant parameters to detect these substances, which particularly affect the development and reproduction of zebrafish. To this end, batches of 30 embryos with the chorion intact were cultured in drinking waters from different sources, throughout their full life-cycle up to 5 months, in 20 L tanks. Six replicates were performed in all water groups, with a total of 24 aquariums. Two generations (F0 and F1) were studied and the following parameters were tested: in the F0 generation, survival and abnormality rates evaluated at 5 dpf (days post-fertilization) and at 5 mpf (months post-fertilization), the onset of spawning and the fertility rate from 3 mpf to 5 mpf, and the sex ratio and underdeveloped specimens at 5 mpf. Furthermore, in the F0 offspring (F1), survival and abnormality rates were evaluated at 5 dpf and the hatching rate at 72 hpf. These results revealed that the hatching rate is the most sensitive parameter to distinguish different levels of effects between waters during the early life stages, whereas the rate of underdeveloped specimens is more suitable at later life stages. Regarding adult reproduction, fertility rate was the most sensitive parameter. The possible reversibility or accumulative nature of such effects will be studied in future work.

Graphene-based biomimetic materials targeting urine metabolite as potential cancer biomarker: application over different conductive materials for potentiometric transduction.

This works presents a novel surface Smart Polymer Antibody Material (SPAM) for Carnitine (CRT, a potential biomarker of ovarian cancer), tested for the first time as ionophore in potentiometric electrodes of unconventional configuration. The SPAM material consisted of a 3D polymeric network created by surface imprinting on graphene layers. The polymer was obtained by radical polymerization of (vinylbenzyl)trimethylammonium chloride and 4-styrenesulfonic acid (signaling the binding sites), and vinyl pivalate and ethylene glycol dimethacrylate (surroundings). Non-imprinted material (NIM) was prepared as control, by excluding the template from the procedure. These materials were then used to produce several plasticized PVC membranes, testing the relevance of including the SPAM as ionophore, and the need for a charged lipophilic additive. The membranes were casted over solid conductive supports of graphite or ITO/FTO. The effect of pH upon the potentiometric response was evaluated for different pHs (2-9) with different buffer compositions. Overall, the best performance was achieved for membranes with SPAM ionophore, having a cationic lipophilic additive and tested in HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer, pH 5.1. Better slopes were achieved when the membrane was casted on conductive glass (-57.4mV/decade), while the best detection limits were obtained for graphite-based conductive supports (3.6×10-5mol/L). Good selectivity was observed against BSA, ascorbic acid, glucose, creatinine and urea, tested for concentrations up to their normal physiologic levels in urine. The application of the devices to the analysis of spiked samples showed recoveries ranging from 91% (± 6.8%) to 118% (± 11.2%). Overall, the combination of the SPAM sensory material with a suitable selective membrane composition and electrode design has lead to a promising tool for point-of-care applications.

Validation of two real-time PCRs targeting the PE-PGRS 20 gene and the region of difference 4 for the characterization of Mycobacterium bovis isolates.

This study aimed to develop and validate real-time PCR for the diagnosis of Mycobacterium bovis isolates. Two hundred and seventy-four M. bovis isolates and 156 M. tuberculosis isolates were tested. Both qPCRs amplified all of the 274 M. bovis samples, but none of the 156 M. tuberculosis samples. The qPCR for PE-PGRS 20 had 91% efficiency and a detection limit of 0.32 ng (sensitivity and specificity for qPCR "Mbovis.100" were 99.64 and 100%, respectively). The qPCR for RD4 had 100% efficiency, and a detection limit of 4 pg (diagnostic sensitivity and specificity were 100 and 100%. The qPCR tests were performed using 4 extraction sets, 3 qPCR kits, and with a range of equipment; yet, all combinations produced similar results in a diagnostic test, demonstrating the robustness of this method. The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.

Chaotic escape of impurities and sticky orbits in toroidal plasmas.

We investigate chaotic impurity transport in toroidal fusion plasmas (tokamaks) from the point of view of passive advection of charged particles due to E×B drift motion. We use realistic tokamak profiles for electric and magnetic fields as well as toroidal rotation effects, and consider also the effects of electrostatic fluctuations due to drift instabilities on particle motion. A time-dependent one degree-of-freedom Hamiltonian system is obtained and numerically investigated through a symplectic map in a Poincaré surface of section. We show that the chaotic transport in the outer plasma region is influenced by fractal structures that are described in topological and metric point of views. Moreover, the existence of a hierarchical structure of islands-around-islands, where the particles experience the stickiness effect, is demonstrated using a recurrence-based approach.

Portable and Autonomous PEDOT-Modified Flexible Paper-Based Methanol Fuel Cell Sensing Platform Applied to L1CAM Recombinant Protein Detection.

This work describes first a 5-stack direct methanol fuel cell (DMFC) based on poly(3,4-ethylenedioxythiophene)-modified paper (PEDOT/PB-DMFC), which acts as an energy source and biosensor, coupled to an electrochromic cell (EC). It is autonomous and monitors the biosensor response by color change, as appropriate for point-of-care (POC) applications. In detail, DMFC strips were developed from square Whatman paper, and the EC was made on baking paper treated with polydimethylsiloxane (PDMS). The PEDOT/PB-DMFCs operate in a passive mode with a few microliters of diluted methanol. The biosensor layer was obtained on the anode ink (a composite of EDOT, oxidized multiwalled carbon nanotubes, and carbon black with platinum and ruthenium) by electropolymerizing 3,4-ethylenedioxythiophene (EDOT), in situ, in the presence of L1CAM. Each PEDOT/PB-DMFC single cell generates a voltage in the range of 0.3-0.35 V depending on the cell, and a five-cell stack delivers a 1.5-1.6 V voltage range when fed with 0.5 M methanol. The fabricated PEDOT/PB-DMFC/biosensor was calibrated against L1CAM, showing linear responses from 1.0 × 10-12 to 1.0 × 10-8 M with a detection limit of 1.17 × 10-13 M (single cell mode). When the EC was connected to the PEDOT/PB-DMFC device, a color gradient was observed. Overall, this work opens horizons to the use of biosensors even in places with energy scarcity and offers an alternative to reducing the current energy demand.

Wave amplitude gain within wedge waveguides through scattering by simple obstacles.

Wave confinement, e.g., in waveguides, gives rise to a huge number of distinct phenomena. Among them, amplitude gain is a recurrent and relevant effect in undulatory processes. Using a general purpose protocol to solve wave equations, the boundary wall method, we demonstrate that for relatively simple geometries, namely, a few leaky or opaque obstacles inside a θ wedge waveguide (described by the Helmholtz equation), one can obtain a considerable wave amplification in certain spatially localized regions of the system. The approach relies on an expression for the wedge waveguide exact Green's function in the case of θ=π/M (M=1,2,...), derived through the method of images allied to group theory concepts. The formula is particularly amenable to numerical calculations, greatly facilitating simulations. As an interesting by-product of the present framework, we are able to obtain the eigenstates of certain closed shapes (billiards) placed within the waveguide, as demonstrated for triangular structures. Finally, we briefly discuss possible concrete realizations for our setups in the context of matter and electromagnetic (for some particular modes and conditions) waves.

Multiparameter DNA content analysis identifies distinct groups in primary breast cancer.

BACKGROUND: Multiparameter flow cytometry is a robust and reliable method for determining tumour DNA content applicable to formalin-fixed paraffin-embedded (FFPE) tissue. This study examined the clinical and pathological associations of DNA content in primary breast cancer using an improved multiparametric technique. METHODS: The FFPE tissue from 201 primary breast cancers was examined and the cancers categorised according to their DNA content using multiparametric flow cytometry incorporating differential labelling of stromal and tumour cell populations. Mathematical modelling software (ModFit 3.2.1) was used to calculate the DNA index (DI) and percentage S-phase fraction (SPF%) for each tumour. Independent associations with clinical and pathological parameters were sought using backward stepwise Binary Logistic Regression (BLR) and Cox's Regression (CR) analysis. RESULTS: Tumours were grouped into four categories based on the DI of the tumour cell population. Low DI tumours (DI=0.76-1.14) associated with progesterone receptor-positive status (P=0.012, BLR), intermediate DI (DI=1.18-1.79) associated with p53 mutant tumours (P=0.001, BLR), high DI (DI1.80) tumours with human epidermal growth factor receptor 2 (HER2)-positive status (P=0.004, BLR) and 'multiploid tumours' (two or more tumour DNA peaks) did not show any significant associations. Tumours with high SPF% (10%) independently associated with poor overall survival (P=0.027, CR). CONCLUSION: Multiparametric flow analysis of FFPE tissue can accurately assess tumour DNA content. Tumour sub-populations associated with biomarkers of prognosis or likely response to therapy. The alterations in DNA content present the potential for greater understanding of the mechanisms underlying clinically significant biomarker changes in primary breast cancer.

Correlation between plasminogen activator inhibitor-1 (PAI-1) promoter 4G/5G polymorphism and metabolic/proinflammatory factors in polycystic ovary syndrome.

Polycystic Ovary Syndrome (PCOS) is the most common cause of subfertility associated to metabolic disorders. The aim of this study was to correlate metabolic and proinflammatory factors in women with PCOS. The frequency of Plasminogen Activator Inhibitor-1 (PAI-1) promoter 4 G/5 G polymorphism was also compared to healthy controls. We evaluated 79 PCOS and 79 healthy women. PAI-1 levels are positively correlated with proinflammatory factors in PCOS group. 4 G allele in PAI-1 gene was more frequent in PCOS and the 4G/4 G genotype was associated with increased PAI-1 levels. A correlation between insulin resistance and proinflammatory and overweight was also observed. C-reactive protein, serum levels of vascular cell adhesion molecule-1 (sVCAM-1), Lipid Accumulation Product (LAP) and vitamin D are good tools to evaluated factors associated to cardiovascular risk in women with PCOS.

Effects of forage type, body condition and single-nucleotide polymorphisms in the bovine cytochrome P450 regulatory region on cow productivity.

Single-nucleotide polymorphisms (SNP) in the coding sequence of cytochrome p450 (CYP3A28) have been associated with milk yield and composition, and calving traits in cows. In this study, we aimed to determine whether (i) the CYP3A28 regulatory region was polymorphic and (ii) SNP genotype, forage type, body condition and their interactions affect cow productivity. Primers for CYP3A28 promoter were designed to amplify a 483-bp segment by PCR. Amplicon sequences revealed seven SNP (T-318C, T-113A, C-189T, T-78G, A6G, G17A and T21C) in Brahman (38 cows), Brahman x Angus reciprocal crosses (47 cows) and crossbreds (98 cows). Angus cows (n = 41) appeared to be fixed at those SNP locations. Genotype and forage {endophyte-infected tall fescue [KY+; Lolium arundinaceum (Schreb.) S. J. Darbyshire] vs. bermudagrass [Cynodon dactylon (L.) Pers.]} effects on lifetime (8-years) calving rate, and calf weaning weights and heights were determined in Herd 1 (126 cows); genotype and BC (low vs. moderate) effects on calving date and calving percent were determined in Herd 2 (98 cows). Four SNP (T-318C, T-113A, A06G and T21C) appeared to be related to cattle productivity, CC cows at T-318C having a lower (p < 0.05) lifetime calving rate than TC or TT cows (65%, 85% and 81% respectively). Cows that grazed KY+ and were TT at T-318C produced calves that tended (p < 0.07) to weigh less than their contemporaries. Moreover, calves of TT cows were shorter (p < 0.05) at weaning than calves of CC or TC cows. In Herd 2, moderate-BC cows that were TT or AA at T-318C, T-113A, T-78G, A6G and T21C had greater (p < 0.05) calving rates (74-80%) than heterozygous cows (46-60%), and low-BC cows that were AA at G17A calved at least 6 days earlier (p < 0.05) than heterozygous cows. Our findings suggest that SNP in the CYP3A28 regulatory region of Brahman-influenced cows are associated with cattle productivity.

New autonomous and self-signaling biosensing device for sarcosine detection.

An early diagnosis is the gold standard for cancer survival. Biosensors have proven their effectiveness in monitoring cancer biomarkers but are still limited to a series of requirements. This work proposes an integrated power solution, with an autonomous and self-signaling biosensing device. The biorecognition element is produced in situ by molecular imprinting to detect sarcosine, a known biomarker for prostate cancer. The biosensor was assembled on the counter-electrode of a dye-sensitized solar cell (DSSC), simultaneously using EDOT and Pyrrole as monomers for the biomimetic process and the catalytic reduction of triiodide in the DSSC. After the rebinding assays, the hybrid DSSC/biosensor displayed a linear behavior when plotting the power conversion efficiency (PCE) and the charge transfer resistance (RCT) against the logarithm of the concentration of sarcosine. The latter obtained a sensitivity of 0.468 Ω/decade of sarcosine concentration, with a linear range between 1 ng/mL and 10 µg/mL, and a limit of detection of 0.32 ng/mL. When interfacing an electrochromic cell, consisting of a PEDOT-based material, with the hybrid device, a color gradient between 1 ng/mL and 10 µg/mL of sarcosine was observed. Thus, the device can be used anywhere with access to a light source, completely equipment-free, suitable for point-of-care analysis and capable of detecting sarcosine within a range of clinical interest.

Determination of microcystin-LR in waters in the subnanomolar range by sol-gel imprinted polymers on solid contact electrodes.

The present work reports new sensors for the direct determination of Microcystin-LR (MC-LR) in environmental waters. Both selective membrane and solid contact were optimized to ensure suitable analytical features in potentiometric transduction. The sensing layer consisted of Imprinted Sol-Gel (ISG) materials capable of establishing surface interactions with MC-LR. Non-Imprinted Sol-Gel (NISG) membranes were used as negative control. The effects of an ionic lipophilic additive, time of sol-gel polymerization, time of extraction of MC-LR from the sensitive layer, and pH were also studied. The solid contact was made of carbon, aluminium, titanium, copper or nickel/chromium alloys (80 : 20 or 90 : 10). The best ISG sensor had a carbon solid contact and displayed average slopes of 211.3 mV per decade, with detection limits of 7.3 × 10(-10) M, corresponding to 0.75 µg L(-1). It showed linear responses in the range of 7.7 × 10(-10) to 1.9 × 10(-9) M of MC-LR (corresponding to 0.77-2.00 µg L(-1)), thus including the limiting value for MC-LR in waters (1.0 µg L(-1)). The potentiometric-selectivity coefficients were assessed by the matched potential method for ionic species regularly found in waters up to their limiting levels. Chloride (Cl(-)) showed limited interference while aluminium (Al(3+)), ammonium (NH(4)(+)), magnesium (Mg(2+)), manganese (Mn(2+)), sodium (Na(+)), and sulfate (SO(4)(2-)) were unable to cause the required potential change. Spiked solutions were tested with the proposed sensor. The relative errors and standard deviation obtained confirmed the accuracy and precision of the method. It also offered the advantages of low cost, portability, easy operation and suitability for adaptation to flow methods.