TC-G 1008 facilitates epileptogenesis by acting selectively at the GPR39 receptor but non-selectively activates CREB in the hippocampus of pentylenetetrazole-kindled mice.

The pharmacological activation of the GPR39 receptor has been proposed as a novel strategy for treating seizures; however, this hypothesis has not been verified experimentally. TC-G 1008 is a small molecule agonist increasingly used to study GPR39 receptor function but has not been validated using gene knockout. Our aim was to assess whether TC-G 1008 produces anti-seizure/anti-epileptogenic effects in vivo and whether the effects are mediated by GPR39. To obtain this goal we utilized various animal models of seizures/epileptogenesis and GPR39 knockout mice model. Generally, TC-G 1008 exacerbated behavioral seizures. Furthermore, it increased the mean duration of local field potential recordings in response to pentylenetetrazole (PTZ) in zebrafish larvae. It facilitated the development of epileptogenesis in the PTZ-induced kindling model of epilepsy in mice. We demonstrated that TC-G 1008 aggravated PTZ-epileptogenesis by selectively acting at GPR39. However, a concomitant analysis of the downstream effects on the cyclic-AMP-response element binding protein in the hippocampus of GPR39 knockout mice suggested that the molecule also acts via other targets. Our data argue against GPR39 activation being a viable therapeutic strategy for treating epilepsy and suggest investigating whether TC-G 1008 is a selective agonist of the GPR39 receptor.

Dietary Zinc Differentially Regulates the Effects of the GPR39 Receptor Agonist, TC-G 1008, in the Maximal Electroshock Seizure Test and Pentylenetetrazole-Kindling Model of Epilepsy.

The G-protein coupled receptor 39 (GPR39) is gaining increasing attention as a target for future drugs, yet there are gaps in the understanding of its pharmacology. Zinc is an endogenous agonist or an allosteric modulator, while TC-G 1008 is a synthetic, small molecule agonist. Zinc is also a positive allosteric modulator for the activity of TC-G 1008 at GPR39. Activation of GPR39 by TC-G 1008 facilitated the development of epileptogenesis in the pentylenetetrazole (PTZ)-induced kindling model of epilepsy. Congruently, TC-G 1008 decreased the seizure threshold in the maximal electroshock seizure threshold (MEST) test. Here, we investigated the effects of TC-G 1008 under the condition of zinc deficiency. Mice were fed a zinc-adequate diet (ZnA, 50 mg Zn/kg) or a zinc-deficient diet (ZnD, 3 mg Zn/kg) for 4 weeks. Following 4 weeks of dietary zinc restriction, TC-G 1008 was administered as a single dose and the MEST test was performed. Additional groups of mice began the PTZ-kindling model during which TC-G 1008 was administered repeatedly and the diet was continued. TC-G 1008 administered acutely decreased the seizure threshold in the MEST test in mice fed the ZnD diet but not in mice fed the ZnA diet. TC-G 1008 administered chronically increased the maximal seizure severity and the percentage of fully kindled mice in those fed the ZnA diet, but not in mice fed the ZnD diet. Our data showed that the amount of zinc in a diet is a factor contributing to the effects of TC-G 1008 in vivo.

Alterations of Serum Magnesium Concentration in Animal Models of Seizures and Epilepsy-The Effects of Treatment with a GPR39 Agonist and Knockout of the Gpr39 Gene.

Several ligands have been proposed for the GPR39 receptor, including the element zinc. The relationship between GPR39 and magnesium homeostasis has not yet been examined, nor has such a relationship in the context of seizures/epilepsy. We used samples from mice that were treated with an agonist of the GPR39 receptor (TC-G 1008) and underwent acute seizures (maximal electroshock (MES)- or 6-hertz-induced seizures) or a chronic, pentylenetetrazole (PTZ)-induced kindling model of epilepsy. MES seizures and PTZ kindling, unlike 6 Hz seizures, increased serum magnesium concentration. In turn, Gpr39-KO mice that underwent PTZ kindling displayed decreased concentrations of this element in serum, compared to WT mice subjected to this procedure. However, the levels of expression of TRPM7 and SlC41A1 proteins-which are responsible for magnesium transport into and out of cells, respectively-did not differ in the hippocampus between Gpr39-KO and WT mice. Furthermore, laser ablation inductively coupled plasma mass spectrometry applied to hippocampal slices did not reveal differences in magnesium levels between the groups. These data show the relationship between magnesium homeostasis and certain types of acute or chronic seizures (MES seizures or PTZ kindling, respectively), but do not explicitly support the role of GPR39 in mediating magnesium balance in the hippocampus in the latter model. However, decreased expression of TRPM7 and increased expression of SLC41A1-which were observed in the hippocampi of Gpr39-KO mice treated with TC-G 1008, in comparison to WT mice that received the same treatment-implicitly support the link between GPR39 and hippocampal magnesium homeostasis.

Association between the Concentrations of Essential and Toxic Elements in Mid-Trimester Amniotic Fluid and Fetal Chromosomal Abnormalities in Pregnant Polish Women.

The present study aimed to investigate the relationship between the concentrations of essential and toxic elements present in the amniotic fluid (AF) and fetal chromosomal abnormalities in pregnant women. A total of 156 pregnant white Polish women aged between 20 and 43 years and screened to detect high risk for chromosomal defects in the first trimester were included in the study. AF samples were collected from these women during routine diagnostic and treatment procedures at mid-gestation (15-22 weeks of their pregnancies). The concentrations of various minerals in the AF were determined by inductively coupled plasma mass spectrometry. Genomic hybridization and cytogenetic karyotyping were performed to detect chromosomal aberrations in the fetuses. The genetic analysis revealed chromosomal aberrations in 19 fetuses (over 12% of all the evaluated women). The major abnormalities identified were trisomy 21 (N = 11), trisomy 18 (N = 2), and triploidy (N = 2). Fetuses with chromosomal abnormalities more frequently showed lower manganese concentration in the AF in the second trimester as compared to those with normal karyotype. A coincidence was observed between high iron levels in the AF and a higher risk of chromosomal abnormalities in the fetuses.

LC/ICP-MS AND COMPLEMENTARY TECHNIQUES IN BESPOKE AND NONTARGETED SPECIATION ANALYSIS OF ELEMENTS IN FOOD SAMPLES.

Chemical elements speciation analysis of food samples has been among the most important scientific topics over the last decades. Food samples are comprised of high variety of chemical compounds, from which many can interact with metals and metalloids, forming complex elemental species with various influence on the human body. It is particularly important not only to determine the amount of certain chemical element in food sample but also to identify the form in which given element occurs in given sample. Employment of bespoke and nontargeted speciation methods, with the use of liquid chromatography inductively coupled plasma mass spectrometry (LC/ICP-MS) and complementary techniques, provides more complete picture on the metals and metalloids speciation in food. This review discusses issues concerning speciation analysis of metals and metalloids in food samples with the use of LC/ICP-MS as a leading technique in elemental speciation nowadays and a complimentary technique intended for their identification. © 2020 John Wiley & Sons Ltd. Mass Spec Rev.

Chemometric approach to find relationships between physiological elements and elements causing toxic effects in herb roots by ICP-MS.

In this paper 13 elements, both physiological and causing toxic effects, were determined by inductively coupled plasma mass spectrometry in roots of 26 species of herbs used in Traditional Chinese Medicine. The herbs were purchased from online shop in two batches 1 year apart to verify the variability of elemental content in time. The multivariate statistical methods-multiple regression, canonical variates and interaction effect analysis-were applied to interpret the data and to show the relationships between elements and two batches of herb roots. The maximum permissible concentration of Cd (0.3 mg kg-1) was exceeded in 7 herb roots which makes 13% of all specimens. The multiple regression analysis revealed the significant relationships between elements: Mg with Sr; V with Pb, As and Ba; Mn with Pb; Fe with As and Ba; Co with Ni and Sr, Cu with Pb, Cd and As; Zn with Pb, Cd, As and Ba. The canonical variates analysis showed that the statistical inference should not be based solely on the type of herb or number of batch because of the underlying interaction effects between those two variables that may be a source of variability of the content of determined elements.

A new procedure for the determination of 21 macro- and trace elements in human fetal urine using an inductively coupled plasma mass spectrometry with dynamic reaction cell (ICP-DRC-MS) equipped with a micro-flow nebulizer.

The procedure for determination of 21 macro- and trace elements - Li, Na, Mg, Al, K, Ca, V, Cr, Mn, Fe, Co, Cu, Zn, Se, Sr, As, Cd, Sb, Ba, Pb and U - in human fetal urine by inductively coupled plasma mass spectrometry (ICP-MS) was developed and validated. The application of a micronebulizer and a dynamic reaction cell (DRC) allowed to perform a full analysis of small volumes (200 µL) of urine collected from human fetuses without the need for sample digestion with closed microwave systems. The procedure and ICP-MS instrument was thoroughly optimized in order to reliably determine both macroelements and ultra-trace concentrations of elements. The internal standard method (Ge, Rh and Tb) was applied in order to encompass signal drift and non-spectral interferences. The rules of metrology were used in order to ensure the quality of the results: (1) the procedure was validated, (2) the uncertainty of the measurement results was estimated and (3) the traceability of the measurement result was established by using the certified reference material with matching matrix (Seronorm Trace Elements Urine L-1). Also, the analyte addition method to the artificial urine was employed for additional confirmation of trueness of the procedure. The selected parameters of the procedure were as follows: (a) limits of detection - (0.00023-53 µg L-1) for U and Ca, respectively, (b) recoveries of the reference value - 81%-136% for Mn and Cd, respectively (c) linearity expressed as R - greater than 0.999, and (d) expanded relative uncertainties (k = 2) - 13%-66% for Sr and Cd, respectively. The developed and validated procedure was applied to 58 samples of urine collected from human fetuses. The samples were diluted with nitric acid and analyzed without further treatment. The procedure allowed to reliably determine both macro- and trace elements in very low volume of sample in a single analytical run.

Total Arsenic and Arsenic Species Determination in Freshwater Fish by ICP-DRC-MS and HPLC/ICP-DRC-MS Techniques.

Analytical methods for the determination of total arsenic (TAs) and arsenic species (arsenite-As(III), arsenate-As(V), monomethylarsenic acid-MMA, dimethylarsenic acid-DMA and arsenobetaine-AsB) in freshwater fish samples were developed. Inductively coupled plasma mass spectrometry with dynamic reaction cell (ICP-DRC-MS) and high-performance liquid chromatography hyphenated to ICP-DRC-MS were used for TAs and arsenic species determination, respectively. The DRC with oxygen as a reaction gas was used. Sample preparation, digestion, and extraction were optimized. Microwave assisted digestion and extraction provided good recovery and extraction efficiency. Arsenic species were fully separated in 8 min using 10 mmol L-1 of ammonium dihydrogen phosphate and 10 mmol L-1 of ammonium nitrate. Overlapping of AsB and As(III) of arsenic species in the presence of a high concentration of AsB and trace amounts of As(III) were studied. Detailed validation of analytical procedures proved the reliability of analytical measurements. Both procedures were characterized by short-term and long-term precision: 2.2% (TAs) up to 4.2% (AsB), and 3.6% (TAs) up to 7.2% (DMA), respectively. Limits of detection (LD) were in the range from 0.056 µg L-1 for TAs to 0.15 µg L-1 for As(V). Obtained recoveries were in the range of 85%⁻116%. Developed methods were applied to freshwater fish samples analysis.

Relationship between pre-pregnancy body mass index and mineral concentrations in serum and amniotic fluid in pregnant women during labor.

The aim of the study was to determine the correlations between body mass index (BMI) values before pregnancy and the concentrations of selected elements (Mg, Co, Cu, Zn, Sr, Cd, Ba, Pb, U, Ca, Cr, Al, Mn, V, Fe) in blood serum and amniotic fluid (AF) in pregnant women. Elemental analysis of serum and amniotic fluid in 225 Polish women (Caucasian/white) showed a relationship between the concentration of minerals in the above-mentioned samples and the pre-pregnancy BMI. Analysis of blood serum was performed by using ICP-MS and it demonstrated that iron concentration was significantly lower in overweight and obese women. Being underweight in pregnant women was associated with a significantly lower concentration of magnesium and cobalt in the blood serum. Both underweight and overweight women were associated with significantly lower concentrations of calcium and strontium in the blood serum. The concentration of cobalt was significantly higher in underweight women. The concentration of lead in the blood serum of overweight and obese women was significantly higher than in other groups. Analysis of the AF showed that the concentration of copper was significantly lower in overweight and obese women, and the concentration of manganese and vanadium significantly higher than in other groups of women. A deficiency in essential minerals and an excess of heavy metals in women may be associated with abnormal body weight and this is important in the etiopathogenesis of pregnancy and fetal development disorders.

Metrological approach to quantitative analysis of clinical samples by LA-ICP-MS: A critical review of recent studies.

Analysis of clinical specimens by imaging techniques allows to determine the content and distribution of trace elements on the surface of the examined sample. In order to obtain reliable results, the developed procedure should be based not only on the properly prepared sample and performed calibration. It is also necessary to carry out all phases of the procedure in accordance with the principles of chemical metrology whose main pillars are the use of validated analytical methods, establishing the traceability of the measurement results and the estimation of the uncertainty. This review paper discusses aspects related to sampling, preparation and analysis of clinical samples by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) with emphasis on metrological aspects, i.e. selected validation parameters of the analytical method, the traceability of the measurement result and the uncertainty of the result. This work promotes the introduction of metrology principles for chemical measurement with emphasis to the LA-ICP-MS which is the comparative method that requires studious approach to the development of the analytical procedure in order to acquire reliable quantitative results.

Usefulness of laser ablation ICP-MS for analysis of metallic particles released to oral mucosa after insertion of dental implants.

Despite the fact that titanium is considered highly biocompatible, its presence in the oral cavity (an environment of frequently changing pH and temperature) may result in the release of titanium from intraosseous implants into the oral mucosa, causing a range of reactions from the human body. Fragments of oral mucosa collected from patients after dental implant insertion were analyzed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The study revealed an elevated content of elements (Ti, Al, V) which are components of the metal implants and temporary cover screws. Dynamic ablation of the tissue surface was used in order to obtain maps of the content and distribution of analyzed elements. The material consisted of 30 oral mucosa tissue fragments collected 3-5 months after implantation and 10 samples collected before implantation (control group). The application of optical microscope allowed for indication and confirmation of the location of metal particles prior to LA-ICP-MS analysis. The so-obtained map permitted location of regions containing metal particles. LA-ICP-MS analysis revealed groups of samples with similar properties of metal particles, thus confirming that those metal particles were the main source of the elevated content of metals (Ti, Al, V) in the tissue after implantation. A calibration strategy based on matrix matched solid standards with powdered egg white proteins as matrix material was applied with 34S as an internal standard. The accuracy of the analytical method was verified by ablating pellets of certified reference material ERM-BB422 Fish muscle.

Influence of stormwater runoff on macroinvertebrates in a small urban river and a reservoir.

The impact of stormwater on benthic macroinvertebrates was studied in two annual cycles. Five small catchments drained by stormwater sewers to a small urban river and a small and shallow reservoir situated in its course were selected. These catchments were located in residential areas with single-family houses or blocks of flats as well as industrial areas, i.e., a car factory, a glassworks and showroom as well as the parking lots of a car dealer and servicing company. In addition to the five stations situated in the vicinity of the stormwater outlets, three stations not directly influenced by stormwater were also established. Macroinvertebrates were sampled in every season, four times per year. Both abundance and biomass were assessed. Stormwater from industrial areas associated with cars, whose catchments showed a high percentage of impervious areas, had the greatest impact on benthic macroinvertebrates. This was due to a large amount of stormwater and its contamination, including heavy metals. Stormwater outflow from residential multi-family houses exerted the least influence. Macroinvertebrates in the water reservoir were found to undergo more extensive changes than those in the river. The cascade of four reservoirs resulted in a marked improvement of water quality in the river, which was confirmed by species composition, abundance and biomass of macroinvertebrates and indicators calculated on their basis for the stations below the cascade in comparison to the stations above and in the first reservoir. These reservoirs replaced constructed wetlands or other measures, which should be undertaken for stormwater management prior to its discharge into urban rivers and other water bodies.

New procedure of quantitative mapping of Ti and Al released from dental implant and Mg, Ca, Fe, Zn, Cu, Mn as physiological elements in oral mucosa by LA-ICP-MS.

A new procedure for determination of elements derived from titanium implants and physiological elements in soft tissues by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is presented. The analytical procedure was developed which involved preparation of in-house matrix matched solid standards with analyte addition based on certified reference material (CRM) MODAS-4 Cormorant Tissue. Addition of gelatin, serving as a binding agent, essentially improved physical properties of standards. Performance of the analytical method was assayed and validated by calculating parameters like precision, detection limits, trueness and recovery of analyte addition using additional CRM - ERM-BB184 Bovine Muscle. Analyte addition was additionally confirmed by microwave digestion of solid standards and analysis by solution nebulization ICP-MS. The detection limits are in range 1.8µgg-1 to 450µgg-1 for Mn and Ca respectively. The precision values range from 7.3% to 42% for Al and Zn respectively. The estimated recoveries of analyte addition line within scope of 83%-153% for Mn and Cu respectively. Oral mucosa samples taken from patients treated with titanium dental implants were examined using developed analytical method. Standards and tissue samples were cryocut into 30µm thin sections. LA-ICP-MS allowed to obtain two-dimensional maps of distribution of elements in tested samples which revealed high content of Ti and Al derived from implants. Photographs from optical microscope displayed numerous particles with µm size in oral mucosa samples which suggests that they are residues from implantation procedure.

Chromium and its speciation in water samples by HPLC/ICP-MS--technique establishing metrological traceability: a review since 2000.

Chromium holds a special position among living organisms because depending on its species it can be either essential or toxic. Cr(VI) even at very low concentrations is harmful and carcinogenic, while Cr(III) is a necessary microelement for cellular metabolism. Therefore, a simple analysis of Cr concentration in collected samples will not be able to distinguish these differences effectively: for a proper chemical analysis we need to perform a reliable detection and quantification of Cr species. Separation and detection of chromium can be accomplished with high performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC/ICP-MS) in a one-step. Our review assembles articles published since 2000 regarding chromium speciation in water samples with the use of HPLC/ICP-MS. It addresses the following issues: chromium chemistry, the possibilities of dealing with interferences, metrological aspects, analytical performance and speciated isotope dilution mass spectrometry (SIDMS) which is a definitive measurement method. The authors would like to advocate this hyphenated advanced technique as well as the metrological approach in speciation analysis of chromium.

Over a century of detection and quantification capabilities in analytical chemistry--historical overview and trends.

The detection limit (LD) and the quantification limit (LQ) are important parameters in the validation process. Estimation of these parameters is especially important when trace and ultra-trace quantities of analyte are to be detected. When the apparatus response from the analyte is below the detection limit, it does not necessarily mean that the analyte is not present in the sample. It may be a message that the analyte concentration could be below the detection capabilities of the instrument or analytical method. By using a more sensitive detector or a different analytical method it is possible to quantitatively determine the analyte in a given sample. The terms associated with detection capabilities have been present in the scientific literature for at least the past 100 years. Numerous terms, definitions and approaches to calculations have been presented during that time period. This paper is an attempt to collect and summarize the principal approaches to the definition and calculation of detection and quantification abilities published from the beginning of 20th century up until the present. Some of the most important methods are described in detail. Furthermore, the authors would like to popularize the knowledge of metrology in chemistry, particularly that part of it which concerns validation of the analytical procedure.