Accumulation ability of trace metals by silver birch leaves in areas contaminated by Zn-Pb ore processing: Effects of excessive trace metal accumulation on specialized metabolism.

The study was carried out in the most industrialised region of southern Poland. Four areas were selected for the study: the vicinity of the Miasteczko Slaskie Zn-Pb-works (ZW; with three transects selected: A, B, and C), and the metalliferous waste heaps (H1, H2, H3). Samples were taken from the surface soil layer, while only mature silver birch (Betula pendula Roth) trees (3-4 m high) growing spontaneously in the study area were selected for the collection of leaves. The analysis of metals including trace metals (TMs; Ag, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Ni, Pb, Tl, and Zn) in the soil and leaves, as well as the contents of specialized leaf compounds belonging to polyphenols were performed. In general, the B. pendula trees in transects A and B located within 500 m of the furnaces exhibited 3-9 times higher content of TMs in their leaves compared to the plants located further away from the Zn smelter. In contrast, the quantity of TM in trees from transect C was independent of distance. The accumulation of TMs in the leaves was significantly lower in the metalliferous waste heap sites compared to the ZW area. It was hypothesised that the accumulation of TMs in B. pendula leaves would depend on the location and distance from the polluting source. It was demonstrated that TMs content in soil affects the accumulation of phenolic acids and flavonoids.

In Vitro Evaluation of Bioavailability of Cr from Daily Food Rations and Dietary Supplements from the Polish Market.

Only some of the nutrients consumed with food are able to be absorbed from the gastrointestinal (GI) tract and enter the systemic circulation (blood). Because some elements are essential minerals for humans, their beneficial effect on the body depends significantly on their bioavailable amount (the fraction that can be absorbed and used by the organism). The term bioavailability, which is very often used to describe the part of nutrients that is able to be absorbed, is influenced by various factors of exogenous and endogenous origin. The main purpose of the study was to assess the relative bioavailability of Cr from selected dietary supplements in the presence of various types of diets, which significantly influence the level of bioavailability. The research was performed using a previously developed and optimized two-stage in vitro digestion model using cellulose dialysis tubes of food rations with the addition of pharmaceutical products. Cr was determined using the ICP-OES and GF-AAS methods, depending on its concentration in particular fractions. The determined relative bioavailability ranged between 2.97 and 3.70%. The results of the study revealed that the type of diet, the chemical form of the molecule, and the pharmaceutical form of preparations have a significant influence on the bioavailability of Cr.

Content of selected heavy metals in the umbilical cord blood and anthropometric data of mothers and newborns in Poland: preliminary data.

The ability to accumulate metals in organs and tissues leads to disturbances in the physiological functioning of the body, causing oxidative stress. This negatively affects the functioning of the placenta and may result in miscarriages, premature birth and fetal growth disorders. The aim of the study was to examine the relationship between the levels of selected heavy metals in umbilical cord blood and anthropometric parameters of mothers and the newborns. Content of elements in umbilical cord blood has been assessed by high-resolution inductively coupled plasma optical emission spectroscopy (ICP-OES). The study results were collected and statistically analyzed using IBM SPSS Statistics software (PS IMAGO). The Pearson correlation coefficient was used to test for associations between selected variables. Regression analysis was conducted to identify predictors of anthropometric parameters of studied women and newborns. The study group consisted of women aged 19-41, whose pregnancy was uncomplicated and were not exposed to heavy metals due to their work or smoking. The following metals were identified in all collected cord blood samples: lead (26.25 ± 9.32 µg/L), zinc (2025.24 ± 717.83 µg/L), copper (749.85 ± 203.86 µg/L), manganese (32.55 ± 13.58 µg/L), chromium (8.34 ± 2.16 µg/L) and selenium (158.46 ± 41.58 µg/L). The conducted statistical analysis indicated the relationship between the copper content in the umbilical cord blood and the weight gain of pregnant women. A significant relationship was observed between newborn head circumference and chromium content. In addition, significant positive correlations were found between the content of zinc and copper, manganese and lead, manganese and selenium, lead and selenium, and lead and chromium in umbilical cord blood. The ratio of zinc to copper concentrations was related to neonatal head circumference. Weight gain in pregnant women is positively correlated with the copper level in umbilical cord blood. There is an association between head circumference at birth and the chromium concentration in umbilical cord blood. Copper and zinc levels in umbilical cord blood are positively correlated with head circumference at birth.

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.

In Vitro Evaluation of Bioavailability of Se from Daily Food Rations and Dietary Supplements.

Bioavailability refers to a fraction of a substance that is potentially absorbed from the gastrointestinal tract and enters the systemic circulation (blood). This term is related to various substances, including minerals, that are present in a complex matrix of food which is consumed every day as natural products and pharmaceutical preparations, e.g., dietary supplements. The purpose of the study was to assess the bioavailability of Se from selected dietary supplements, with the simultaneous assessment of the effect the diet type (standard, basic and high-residue diets) has on relative bioavailability. The research included a two-stage in vitro model of digestion using cellulose dialysis tubes of the food rations with the addition of dietary supplements. Se was determined using the ICP-OES method. The bioavailability of Se from dietary supplements, in the presence of food matrix, was determined to be within the range of 19.31-66.10%. Sodium selenate was characterized by the highest value of this parameter, followed by organic forms and sodium selenite. The basic diet, characterized by moderate protein and high carbohydrate and fiber contents, positively influenced the bioavailability of Se. The bioavailability of Se was also influenced by the pharmaceutical form of the product-the highest was for tablets, followed by capsules and coated tablets.

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.

Is there any direct link between hazardous trace metals and the allantoin content in some moss species?

The accumulation of allantoin and trace metals (TMs) in nine moss species was examined after the exposure to stress conditions. Both the environmental anthropopressure effect and laboratory-simulated stress conditions were monitored. Moss samples were collected from different locations, i.e. a non-TM contaminated area, an urban area, and a metalliferous area. The effect of Cd, Pb, Hg, Ni, Zn, salinity, and an acidic environment on the allantoin content was tested. Principal component analysis was performed to reveal the relationship between samples of different origin. Large differences in the metal and allantoin accumulation capability of mosses were noted between samples harvested from the different locations. Seven species were considered as potential metal accumulators, as they exhibited tolerance to elevated levels of heavy metals. The observed TM effect on the allantoin accumulation indicated TM pollution as an important environmental factor that can significantly influence the content of this compound in mosses. Further studies on the contribution of various environmental factors and individual characteristics of plant species are highly expected to recognize the trend in the accumulation of specialized metabolites and TMs in response to hazardous growth conditions.

Reactivation of Vertebral Growth Plate Function in Vertebral Body Tethering in an Animal Model.

Flexible spine tethering is a relatively novel fusionless surgical technique that aims to correct scoliosis based on growth modulation due to the pressure exerted on the vertebral body epiphyseal growth plate. The correction occurs in two phases: immediate intraoperative and postoperative with growth. The aim of this study was to evaluate the reactivation of vertebral growth plate function after applying corrective forces. The rat tail model was used. Asymmetric compression and distraction of caudal growth plates were performed using a modified external fixation apparatus. Radiological and histopathological data were analysed. After three weeks of correction, the activity of the structures increased across the entire growth plate width, and the plate was thickened. The height of the hypertrophic layer and chondrocytes on the concave side doubled in height. The height of chondrocytes and the cartilage thickness on the concave and central sides after the correction did not differ statistically significantly from the control group. Initiation of the correction of scoliosis in the growing spine, with relief of the pressure on the growth plate, allows the return of the physiological activity of the growth cartilage and restoration of the deformed vertebral body.

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.

Some Natural Photosensitizers and Their Medicinal Properties for Use in Photodynamic Therapy.

Despite significant advances in early diagnosis and treatment, cancer is one of the leading causes of death. Photodynamic therapy (PDT) is a therapy for the treatment of many diseases, including cancer. This therapy uses a combination of a photosensitizer (PS), light irradiation of appropriate length and molecular oxygen. The photodynamic effect kills cancer cells through apoptosis, necrosis, or autophagy of tumor cells. PDT is a promising approach for eliminating various cancers but is not yet as widely applied in therapy as conventional chemotherapy. Currently, natural compounds with photosensitizing properties are being discovered and identified. A reduced toxicity to healthy tissues and a lower incidence of side effects inspires scientists to seek natural PS for PDT. In this review, several groups of compounds with photoactive properties are presented. The use of natural products has been shown to be a fruitful approach in the discovery of novel pharmaceuticals. This review focused on the anticancer activity of furanocoumarins, polyacetylenes, thiophenes, tolyporphins, curcumins, alkaloid and anthraquinones in relation to the light-absorbing properties. Attention will be paid to their phototoxic and anti-cancer effects on various types of cancer.

Analytical Problems with Preparation of Paraspinal Tissues from Patients with Spinal Fusion for Analysis of Titanium.

Preparation of paraspinal tissue of patients with implants for elemental analysis is a challenge because it contains titanium in the ionic form, as well as metallic debris. Most literature reports focus on dissolving the tissue, but the impact of digestion conditions on metallic debris of Ti has not been investigated. In our work, various digestion conditions, including systems, compositions of oxidising mixture, and time, were tested aiming (i) to digest the tissue without digestion of metallic titanium to quantify soluble Ti and (ii) to digest metallic titanium debris to asses total Ti content in tissue. The experiments were performed in a closed mode using a microwave-assisted system and a carbon heating block. Our study revealed that total digestion of titanium was impossible in the tested conditions and the maximal level of digested titanium was below 70%. The mineralisation with the use of concentrated nitric acid was optimal to prepare paraspinal samples to analyse the soluble titanium form because metallic titanium passivated and did not migrate to the solution. The elaborated conditions were applied to determine titanium ion in the periimplant tissue of patients with three different titanium-based surgical systems, including traditional growing rod (TGR), guided growth systems (GGS), and vertical expandable prosthesis titanium rib (VEPTR).

Comparison of Different Surgical Systems for Treatment of Early-onset Scoliosis in the Context of Release of Titanium Ions.

STUDY DESIGN: Case-control study. OBJECTIVE: The aim of the study was to compare growth-friendly systems: traditional growing rod, guided growth systems (GGS), and vertical expandable prosthetic titanium rib in the context of titanium release. SUMMARY OF BACKGROUND DATA: The problem of scoliosis affects even up to 3% of the population, and ca. 0.1% of patients need surgical treatment. Surgical treatment carries the risk of a long-term presence of implants in the organism, which may result in release of metal ions into the tissues and bloodstream. METHODS: Seventy-one patients (13.5 ±â€Š3.54 years' old) were treated for spinal deformity using various surgical systems and the samples of paraspinal tissue, blood, nails, and hair were collected before and after treatment. The quantification of titanium was performed using inductively coupled plasma optical emission spectrometry. RESULTS: The metallic particles were released into the peri-implant tissue, and the greatest amounts of titanium were detected in patients with GGS. The concentration of soluble titanium forms in subcutaneous tissue (ST) was low and do not statistically differ from control. The average titanium content in the paraspinal tissue in patients with GGS was two- to three-fold higher than the average value in the other investigated groups. A slightly increased level of titanium compared with the control was noted in all studied groups of patients. The highest content of titanium in blood was observed in patients with the GGS system. CONCLUSION: Neverless the system used, the concentration of soluble titanium forms in both ST and blood was only slightly higher than in the control and did not exceed the allowable levels. The increased level of titanium with GGS system is probably associated with the friction between implant components, whereas the components in the other systems are immobile relative to each other.Level of Evidence: 3.

Morphological, Anatomical, and Phytochemical Studies of Carlina acaulis L. Cypsela.

Carlina acaulis L. has a long tradition of use in folk medicine. The chemical composition of the roots and green parts of the plant is quite well known. There is the lowest amount of data on the cypsela (fruit) of this plant. In this study, the microscopic structures and the chemical composition of the cypsela were investigated. Preliminary cytochemical studies of the structure of the Carlina acaulis L. cypsela showed the presence of substantial amounts of protein and lipophilic substances. The chemical composition of the cypsela was investigated using spectrophotometry, gas chromatography with mass spectrometry, and high-performance liquid chromatography with spectrophotometric and fluorescence detection. The cypsela has been shown to be a rich source of macro- and microelements, vegetable oil (25%), α-tocopherol (approx. 2 g/kg of oil), protein (approx. 36% seed weight), and chlorogenic acids (approx. 22 g/kg seed weight). It also contains a complex set of volatile compounds. The C. acaulis cypsela is, therefore, a valuable source of nutrients and bioactive substances.

Tolerance of Facultative Metallophyte Carlina acaulis to Cadmium Relies on Chelating and Antioxidative Metabolites.

The impact of long-term chronic cadmium stress (ChS, 0.1 µM Cd, 85 days) or short-term acute cadmium stress (AS, 10 µM Cd, 4 days) on Carlina acaulis (Asteraceae) metabolites was compared to identify specific traits. The content of Cd was higher under AS in all organs in comparison with ChS (130 vs. 16 µg·g-1 DW, 7.9 vs. 3.2 µg·g-1 DW, and 11.5 vs. 2.4 µg·g-1 DW in roots, leaves, and trichomes, respectively) while shoot bioaccumulation factor under ChS (ca. 280) indicates efficient Cd accumulation. High content of Cd in the trichomes from the AS treatment may be an anatomical adaptation mechanism. ChS evoked an increase in root biomass (hormesis), while the impact on shoot biomass was not significant in any treatment. The amounts of ascorbic acid and sum of phytochelatins were higher in the shoots but organic (malic and citric) acids dominated in the roots of plants from the ChS treatment. Chlorogenic acid, but not ursolic and oleanolic acids, was elevated by ChS. These data indicate that both chelation and enhancement of antioxidative power contribute to protection of plants exposed to long-term (chronic) Cd presence with subsequent hormetic effect.

Applicability of a Monolithic Column for Separation of Isoquinoline Alkalodis from Chelidonium majus Extract.

Isoquinoline alkaloids are the main group of secondary metabolites present in Chelidonium majus extracts, and they are still the object of interest of many researchers. Therefore, the development of methods for the investigation and separation of the alkaloids is still an important task. In this work, the application potential of a silica-based monolithic column for the separation of alkaloids was assessed. The influence of the organic modifier, temperature, salt concentration, and pH of the eluent on basic chromatographic parameters such as retention, resolution between neighboring peaks, chromatographic plate numbers, and peak asymmetry were investigated. Based on the obtained results, a gradient elution program was developed and used to separate and quantitatively determine the main alkaloids in a Chelidonium majus root extract.

Polyaniline based material as a new SPE sorbent for pre-treatment of Chelidonium majus extracts before chromatographic analysis of alkaloids.

In the present study, deprotonated and protonated polyaniline based sorbent obtained by in situ polymerization of aniline on silica (Si-PANI) was applied for SPE pretreatment of Chelidonium majus extracts before determination of benzophenanthridine, protoberberine and protopine alkaloids. The experimentally optimized conditions, such as protonated polyaniline impregnated with methanol as a sorbent, 2 mL of water and methanol mixture (1/1, v/v) as a washing solution, and 5 mL of 0.1 M methanolic solution of ammonia as an elution solvent, yielded the highest values of analyte recovery (above 96.5%) with simultaneous removal of undesirable plant matrix. Virtually modeled structure of polyaniline revealed possible occurrence of the mixed mode binding mechanism, based both on π-π and ion interactions on protonated form of polyaniline. Moreover, lack of significant changes of the polyaniline film quality assessed by Raman spectroscopy after series of ten experiments proved the reusability of the sorbent.

Effect of Long-Term Strontium Exposure on the Content of Phytoestrogens and Allantoin in Soybean.

Abiotic stress, including metal excess, can modify plant metabolism. Here we investigated the influence of long-term strontium exposure (12 weeks, 0.5⁻4.0 mM Sr) on the content of phytoestrogens and allantoin as well as the mineral composition in soybean. Seven phytoestrogens were identified in the soybean: daidzin, glycitin, genistin, malonyldaidzin, malonylgenistin, daidzein, and coumestrol. The results showed that both malonyldaidzin and malonylgenistin were dominant phytoestrogens; however, the roots contained a relatively high amount of daidzein. It was found that strontium reduced the phytoestrogen content and decreased the antioxidant capacity. Strontium evoked depletion of the sum of all phytoestrogens by 40⁻70% in the leaves, 25⁻50% in the stems and in the seeds, depending on the strontium concentration. In the roots, 0.5 and 4.0 mM of strontium decreased the total phytoestrogen content by 25 and 55%, respectively, while 2.0 mM of strontium did not exert an effect on their accumulation. On the other hand, strontium ions induced allantoin accumulation mainly in the roots. Strontium was preferentially accumulated in the leaves, with a slight impact on macro- and micro-nutrients. Our research showed strontium-secondary metabolites interaction in the soybean, which can be useful for obtaining a natural pharmaceutical product containing both strontium and phytoestrogens for remediation of postmenopausal osteoporosis.

Systematic Evaluation of Chromatographic Parameters for Isoquinoline Alkaloids on XB-C18 Core-Shell Column Using Different Mobile Phase Compositions.

Chelidonium majus L. is a rich source of isoquinoline alkaloids with confirmed anti-inflammatory, choleretic, spasmolytic, antitumor, and antimicrobial activities. However, their chromatographic analysis is difficult because they may exist both in charged and uncharged forms and may result in the irregular peak shape and the decrease in chromatographic system efficacy. In the present work, the separation of main C. majus alkaloids was optimized using a new-generation XB-C18 endcapped core-shell column dedicated for analysis of alkaline compounds. The influence of organic modifier concentration, addition of salts, and pH of eluents on chromatographic parameters such as retention, resolution, chromatographic plate numbers, and peak asymmetry was investigated. The results were applied to elaborate the optimal chromatographic system for simultaneous quantification of seven alkaloids from the root, herb, and fruit of C. majus.

Silica Modified with Polyaniline as a Potential Sorbent for Matrix Solid Phase Dispersion (MSPD) and Dispersive Solid Phase Extraction (d-SPE) of Plant Samples.

Polyaniline (PANI) is one of the best known conductive polymers with multiple applications. Recently, it was also used in separation techniques, mostly as a component of composites for solid-phase microextraction (SPME). In the present paper, sorbent obtained by in situ polymerization of aniline directly on silica gel particles (Si-PANI) was used for dispersive solid phase extraction (d-SPE) and matrix solid-phase extraction (MSPD). The efficiency of both techniques was evaluated with the use of high performance liquid chromatography with diode array detection (HPLC-DAD) quantitative analysis. The quality of the sorbent was verified by Raman spectroscopy and microscopy combined with automated procedure using computer image analysis. For extraction experiments, triterpenes were chosen as model compounds. The optimal conditions were as follows: protonated Si-PANI impregnated with water, 160/1 sorbent/analyte ratio, 3 min of extraction time, 4 min of desorption time and methanolic solution of ammonia for elution of analytes. The proposed procedure was successfully used for pretreatment of plant samples.

Proliferative and antioxidant activity of Symphytum officinale root extract.

The root of Symphytum officinale L. is commonly used in folk medicine to promote the wound healing, reduce the inflammation and in the treatment of broken bones. The objective of our investigation was to analyse the extract from S. officinale in term of its antioxidant activity and the effect on cell viability and proliferation of human skin fibroblast (HSF). Moreover, the quantification of main phenolics and allantoin was conducted using HPLC-DAD method. Five compounds were found: rosmarinic, p-hydroxybenzoic, caffeic, chlorogenic and p-coumaric acid. DPPH, FRAP and TPC assay showed the high antioxidant activity of the extract. MTT test proved the stimulatory effect on cell metabolism and viability of HSF cells. Moreover, no changes in cytoskeleton structure and cells shape were observed. The obtained results indicate that non-toxic extract from S. officinale root has strong antioxidant potential and a beneficial effect on human skin fibroblasts.