Dynamics of Serum Inflammatory Markers and Adipokines in Patients: Implications for Monitoring Abnormal Body Weight: Preliminary Research.

This study aimed to investigate the influence of abnormal body weight on inflammatory markers and adipokine levels across varied body mass index (BMI) categories. The cohort included 46 participants categorized into normal BMI (group I; n = 19), overweight (group II; n = 14), and obesity (group III; n = 13). Inflammatory markers (hsCRP and IL-6) and adipokines (Adiponectin, Leptin, Nesfatin-1, and Zinc-α2-glycoprotein) were assessed to discern effective indicators of inflammation in individuals with abnormal body weight. Additionally, the full lipid profile was also assessed (total cholesterol, triglycerides, LDL-C, HDL-C). The results indicated significant biochemical changes, particularly in IL-6 and Leptin levels, in participants with a BMI over 25. The levels of ZAG protein were negatively correlated with the HDL-C and LDC-L levels with statistical significance (Pearson: -0.57, p = 0.001, and Pearson: -0.41, p = 0.029, for HDL-C and LDL-C, respectively), suggesting that the level of ZAG is also inversely proportional to the amount of cholesterol. Statistical analyses revealed decreased Zinc-α2-glycoprotein (ZAG) levels and increased Adiponectin, Leptin, and IL-6 levels in individuals with abnormal body weight. Correlation analyses demonstrated a statistically significant upward trend for IL-6 (p = 0.0008) and Leptin (p = 0.00001), with a similar trend observed for hsCRP without statistical significance (p = 0.113). IL-6 levels in the overweight group were 158.71% higher than in the normal-weight group, while the obese group exhibited a 229.55% increase compared to the normal-weight group. No notable changes have been recorded for the levels of Nesfatin-1. Based on our results, we propose IL-6, Leptin, and ZAG as potential biomarkers for monitoring interventions and assessing patient conditions in those with abnormal BMIs. Further research with a larger patient cohort is warranted to validate these correlations in overweight and obese individuals.

Potential Role of IL-37 in Atopic Dermatitis.

Interleukin 37 (IL-37) is a recently discovered member of the IL-1 cytokine family that appears to have anti-inflammatory and immunosuppressive effects in various diseases. IL-37 acts as a dual-function cytokine, exerting its effect extracellularly by forming a complex with the receptors IL-18 α (IL-18Rα) and IL-1R8 and transmitting anti-inflammatory signals, as well as intracellularly by interacting with Smad3, entering the nucleus, and inhibiting the transcription of pro-inflammatory genes. Consequently, IL-37 is linked to IL-18, which plays a role in the pathogenesis of atopic dermatitis (AD), consistent with our studies. Some isoforms of IL-37 are expressed by keratinocytes, monocytes, and other skin immune cells. IL-37 has been found to modulate the skewed T helper 2 (Th2) inflammation that is fundamental to the pathogenesis of AD. This review provides an up-to-date summary of the function of IL-37 in modulating the immune system and analyses its potential role in the pathogenesis of AD. Moreover, it speculates on IL-37's hypothetical value as a therapeutic target in the treatment of AD.

Effect of Manual Lymphatic Drainage on the Concentrations of Selected Adipokines, Cytokines, C-Reactive Protein and Parameters of Carbohydrate and Lipid Metabolism in Patients with Abnormal Body Mass Index: Focus on Markers of Obesity and Insulin Resistance.

The aim of the study was to assess the impact of manual lymphatic drainage (MLD) on the parameters of carbohydrate metabolism, lipid metabolism and the level of selected adipokines and cytokines in people with abnormal body mass index (BMI). In addition, an attempt was made to assess the optimal cut-off values of serum concentrations of the biochemical parameters studied in identifying the risk of obesity and insulin resistance (IR). The study included 60 subjects who underwent 10 and 30 min long MLD sessions three times a week. The study group included 15 patients with a normal body mass index (group I; n = 15), overweight patients (group II; n = 15) and obese patients (group III; n = 10). The control group was IV; n = 20 subjects not undergoing MLD. Biochemical tests were carried out on all subjects at stage 0' (before MLD therapy) and at stage 1' (one month after MLD therapy). In the control group, the time between the sample collection at stage 0' and stage 1' was the same as in the study group. Our results showed that 10 MLD sessions may have a positive effect on the selected biochemical parameters, including insulin, 2h-PG, leptin and HOMA-IR values in normal weight and overweight patients. In addition, in the study group, the highest AUCROC values in identifying the risk of obesity were found for leptin (AUCROC = 82.79%; cut-off = 17.7 ng/mL; p = 0.00004), insulin (AUCROC = 81.51%; cut-off = 9.5 µIU/mL; p = 0.00009) and C-peptide (AUCROC = 80.68%; cut-off = 2.3 ng/mL; p = 0.0001) concentrations as well as for HOMA-IR values (AUCROC = 79.97%; cut-off = 1.8; p = 0.0002). When considering the risk of IR, we observed the highest diagnostic value for insulin (AUCROC = 93.05%; cut-off = 1.8 ng/mL; p = 0.053), which was followed by C-peptide (AUCROC = 89.35%; cut-off = 17.7 ng/mL; p = 0.000001), leptin (AUCROC = 79.76%; cut-off = 17.6 ng/mL; p = 0.0002) and total cholesterol (AUCROC = 77.31%; cut-off = 198 mg/dL; p = 0.0008). Our results indicate that MLD may have a positive effect on selected biochemical parameters, including insulin, 2h-PG, leptin and HOMA-IR, in normal weight and overweight patients. In addition, we successfully established optimal cut-off values for leptin in the assessment of obesity and insulin in the assessment of insulin resistance in patients with abnormal body mass index. Based on our findings, we hypothesize that MLD, when combined with caloric restriction and physical activity, may serve as an effective preventive intervention against the development of obesity and insulin resistance.

Impact of hydration with beverages containing free sugars or xylitol on metabolic and acute kidney injury markers after physical exercise.

The proper fluid and carbohydrates intake is essential before and during physical exercise, and for this reason most athletes drink beverages containing a high amount of free sugars. Sweetened soft drinks are also commonly consumed by those not doing any sport, and this habit seems to be both unhealthy and also the cause of metabolic problems. Recently, several sweeteners have been proposed to replace sugars in popular beverages. To examine the impact of free sugars and the popular sweetener xylitol on metabolic profile and the markers of kidney function and injury after exercise the present study was conducted with semi-professional football players. All participants were healthy, with a mean age of 21.91 years. Their sports skills were on the level of the 4th-5th division of the league. The subjects took part in four football training sessions. During each session they drank a 7% solution of sugar (sucrose, fructose, glucose) or xylitol. The tolerability of these beverages and well-being during exercise was monitored. Before and after each training session, blood and urine were collected. The markers of kidney function and injury, uric acid, electrolytes, complete blood count, CRP, serum albumin, serum glucose and the lipid profile were analyzed. The main finding of this study was that the xylitol beverage is the least tolerated during exercise and 38.89% of participants experienced diarrhea after training and xylitol intake. Xylitol also led to unfavorable metabolic changes and a large increase in uric acid and creatinine levels. A mean increase of 1.8 mg/dl in the uric acid level was observed after xylitol intake. Increases in acute kidney injury markers were observed after all experiments, but changes in urine albumin and cystatin C were highest after xylitol. The other three beverages (containing "free sugars" - glucose, fructose and sucrose) had a similar impact on the variables studied, although the glucose solution seems to have some advantages over other beverages. The conclusion is that sweeteners are not a good alternative to sugars, especially during exercise. Pure water without sweeteners should be drunk by those who need to limit their calorie consumption. Clinical Trial Registration: ClinicalTrials.gov, (NCT04310514).

Data on the identification of microsatellite markers in Eisenia fetida and Eisenia andrei.

Eisenia fetida and Eisenia andrei are closely related earthworm species that play a crucial part in soil and influence its structure and organic matter cycling. Due to their essential environmental role, they are widely used as model organisms in a vast spectrum of research areas. In this work, we partially sequenced genomes of E. fetida and E. andrei, using Illumina technology (Nano 2 × 250 v2 - MiSeq) and de novo assembly strategy. A total of 3785 and 4258 microsatellite or Simple Sequence Repeat (SSR) markers were identified within E. fetida and E. andrei genomic DNA, respectively. The microsatellite markers will facilitate the analyses of genetic diversity and population genetics studies for the two selected earthworm species and their interspecific hybrids.

Air Pollution, Oxidative Stress, and the Risk of Development of Type 1 Diabetes.

Despite multiple studies focusing on environmental factors conducive to the development of type 1 diabetes mellitus (T1DM), knowledge about the involvement of long-term exposure to air pollution seems insufficient. The main focus of epidemiological studies is placed on the relationship between exposure to various concentrations of particulate matter (PM): PM1, PM2.5, PM10, and sulfur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (O3), versus the risk of T1DM development. Although the specific molecular mechanism(s) behind the link between increased air pollution exposure and a higher risk of diabetes and metabolic dysfunction is yet unknown, available data indicate air pollution-induced inflammation and oxidative stress as a significant pathway. The purpose of this paper is to assess recent research examining the association between inhalation exposure to PM and associated metals and the increasing rates of T1DM worldwide. The development of modern and more adequate methods for air quality monitoring is also introduced. A particular emphasis on microsensors, mobile and autonomous measuring platforms, satellites, and innovative approaches of IoT, 5G connections, and Block chain technologies are also presented. Reputable databases, including PubMed, Scopus, and Web of Science, were used to search for relevant literature. Eligibility criteria involved recent publication years, particularly publications within the last five years (except for papers presenting a certain novelty or mechanism for the first time). Population, toxicological and epidemiological studies that focused particularly on fine and ultra-fine PM and associated ambient metals, were preferred, as well as full-text publications.

Significant Decrease in Glycated Hemoglobin, 2h-Post-Load Glucose and High-Sensitivity C-Reactive Protein Levels in Patients with Abnormal Body Mass Index after Therapy with Manual Lymphatic Drainage.

The objective of this study was to investigate the effect of manual lymphatic drainage (MLD) on the insulin resistance parameter (HOMA-IR), glycated hemoglobin (HbA1c), C-peptide, insulin, fasting plasma glucose (FPG), 2h-post-loadglucose (2h-PG) and the concentration of high-sensitivity C-reactive protein (hsCRP) in patients with abnormal body mass index. The study involved 30 patients, including patients with normal body weight (as a control group; group I; n = 14), overweight patients (group II; n = 9) and obese patients (group III; n = 7). Each patient underwent 10 sessions of MLD therapy, 3 times a week for 30 min. In addition, we measured body mass index (BMI) and waist-to-hip ratio (WHR) and performed body composition analysis as well as biochemical tests before MLD therapy (stage 0') and after MLD therapy (stage 1'). A statistically significant correlation was demonstrated between the concentration of C-peptide, BMI, the amount of visceral adipose tissue (r = 0.87, p = 0.003; r = 0.76, p = 0.003, respectively), and the HOMA-IR index, BMI and the amount of visceral adipose tissue (r = 0.86, p = 0.005; r = 0.84, p = 0.042, respectively), before and after MLD therapy. In overweight patients (group II), a statistically significant (p = 0.041) decrease in the hsCRP level by 2.9 mg/L and a significant (p = 0.050) decrease in the 2h-PG level by 12 mg/dL after the MLD therapy was detected. Moreover, in the group of obese patients (group III), a statistically significant (p = 0.013) decrease in HbA1c level by 0.2% after MLD therapy was demonstrated. Our results indicate that MLD may have a positive effect on selected biochemical parameters, with the most favorable changes in overweight patients. Further studies in a larger number of patients are warranted to confirm our findings, to test in-depth their mechanism, and to investigate clinical benefits of this alternative therapy in patients with abnormal body mass index.

Effect of Manual Therapy Compared to Ibuprofen on Primary Dysmenorrhea in Young Women-Concentration Assessment of C-Reactive Protein, Vascular Endothelial Growth Factor, Prostaglandins and Sex Hormones.

BACKGROUND: The study aimed to assess if manual therapy, compared to ibuprofen, impacts the concentration of inflammatory factors, sex hormones, and dysmenorrhea in young women Methods: Thirty-five women, clinically diagnosed with dysmenorrhea, were included in the study. They were divided into group A-manual therapy (n = 20) and group B-ibuprofen therapy (n = 15). Inflammatory factors such as vascular endothelial growth factor (VEGF), C-reactive protein (CRP), prostaglandin F2α (PGF2α), E2 (PGE2) and sex hormones levels were measured. Dysmenorrhea assessed with the numerical pain rating scale (NPRS), myofascial trigger points, and muscle flexibility were examined before and after the interventions. RESULTS: The difference in the level of 17-ß-estradiol after manual and ibuprofen therapy was significant, as compared to baseline (p = 0.036). Progesterone levels decreased in group A (p = 0.002) and B (p = 0.028). The level of CRP was negatively correlated with sex hormones. Decrease in dysmenorrhea was significant in both groups (group A p = 0.016, group B p = 0.028). Non-significant differences were reported in prostaglandins, VEGF and CRP levels, in both groups. CONCLUSIONS: There were no significant differences in CRP, prostaglandins and VEGF factors after manual or ibuprofen therapy. It has been shown that both manual therapy and ibuprofen can decrease progesterone levels. Manual therapy had a similar effect on the severity of dysmenorrhea as ibuprofen, but after manual therapy, unlike after ibuprofen, less muscles with dysfunction were detected in patients with primary dysmenorrhea.

Environmental Factors and the Risk of Developing Type 1 Diabetes-Old Disease and New Data.

The incidence of type 1 diabetes (T1D) is increasing worldwide. The onset of T1D usually occurs in childhood and is caused by the selective destruction of insulin-producing pancreatic islet cells (ß-cells) by autoreactive T cells, leading to insulin deficiency. Despite advanced research and enormous progress in medicine, the causes of T1D are still not fully understood. Therefore, an extensive online search for scientific research on environmental factors associated with diabetes and the identification of new factors of unexplained etiology has been carried out using the PubMed, Cochrane, and Embase databases. The search results were limited to the past 11 years of research and discovered 143 manuscripts published between 2011 and 2022. Additionally, 21 manuscripts from between 2000 and 2010 and 3 manuscripts from 1974 to 2000 were referenced for historical reference as the first studies showcasing a certain phenomenon or mechanism. More and more scientists are inclined to believe that environmental factors are responsible for the increased incidence of diabetes. Research results show that higher T1D incidence is associated with vitamin D deficiency, a colder climate, and pollution of the environment, as well as the influence of viral, bacterial, and yeast-like fungi infections. The key viral infections affecting the risk of developing T1DM are rubella virus, mumps virus, Coxsackie virus, cytomegalovirus, and enterovirus. Since 2020, i.e., from the beginning of the COVID-19 pandemic, more and more studies have been looking for a link between Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and diabetes development. A better understanding of the role of viral, bacterial, and yeast-like fungi infections related to the risk of T1DM in children and adolescents and the identification of new risk factors, especially those spread by the droplet route, is of great importance for people and families with diabetes.

Transcriptome sequencing of Brassica napus highlights the complex issues with soil supplementation with sewage sludge.

The soil supplementation with sewage sludge (SS) has become a widespread method to improve soil quality, but its long-term possible consequences are still relatively unknown. SS may contain several groups of contaminants to which the biological responses of the organisms are still poorly understood mainly due to the mixture toxicity. In this context, RNA-seq has been used to assess the impact of the exposure to sewage sludge supplemented soil at the whole-transcriptome level in the Brassica napus (B. napus). Although the municipal sewage sludge passed all safety regulations set by the EU commission (86/278/EEC), soil supplementation with SS caused a significant (p < 0.05) increase in the content of lead (by 68.8%, 71.4% in plant shoots and roots, respectively), zinc (by 22.4% and 31.2%), nickel (by 67.0% and 30.2%), and copper (by 33.1% and 39.2%). The de-novo assembled transcriptome of B. napus identified 555 differently expressed genes (DEGs) in a response to sewage sludge supplementation at the false detection rate below 0.001 (FDR <0.001). Among them, 313 genes were up-regulated and 242 genes were down-regulated. The gene ontology analysis (GO) had shown, that significantly enriched GO groups included genes involved in photosynthesis, carbohydrate metabolism and photosystems repair (41 genes), response to oxidative stress (50 genes), response to pathogens (36 genes), response to xenobiotics (15 genes), and heavy metals (41 genes), cell death (8 genes), cell wall structure (15 genes). These results suggest a significant impact of contaminants in sewage sludge on plants transcriptome. The transcriptomic approach facilitated a better understanding of the molecular level of the potential toxicity of sewage sludge in B. napus. RNA-seq allowed for the identification of potential novel early-warning molecular markers of environmental contamination. This work highlights the crucial necessity for rapid legislation change concerning the allowable levels of contaminants in sewage sludge applied on land, to mitigate the possible adverse outcomes in the ecosystem after its use as a fertilizer.

The Effects of Wastewater Treatment Plant Failure on the Gulf of Gdansk (Southern Baltic Sea).

In August 2019 and during August/September 2020, the main collection system of the Wastewater Treatment Plant (WWTP) in Warsaw, Poland, malfunctioned. During that system failure, over 4.8 million m3 of untreated wastewater was dropped directly into the Vistula River in just a few days. It is currently considered as one of the largest known failures of WWTP worldwide. In order to assess the environmental impact, water samples were collected from 2 spots at the Vistula river estuary (406 and 415 km from the discharge location, respectively), and 4 spots at the Gulf of Gdansk, situated on the southern shore of the Baltic Sea. The sampling was conducted before the wastewater wave reached the Vistula river's mouth, followed by daily sampling during 21 days after the malfunction occurred. The study showed the decline in water quality at the Vistula river estuary and the Baltic shore waters as the wave of wastewater reached those points, despite being situated over 400 km downstream from the place of the accident. Those changes included the reduction in the dissolved oxygen content (by 0.69-fold at its peak), the increase in Total Organic Carbon (TOC) (by 1.28-fold at its peak), nitrate-nitrogen (N-NO3) (by 1.68-fold at its peak), phosphorous (P) (by 2.41-fold at its peak), conductivity (by 16.8-fold at its peak), and Chemical Oxygen Demand (COD) (by 1.84-fold). In the samples from the Vistula river, the decline in water quality was seen as incidental and lasted 2-3 days. Subsequently, the levels of physical and chemical parameters returned to the levels from before the accident. However, the changes in the Gulf of Gdansk lasted significantly longer, especially on the West side of the Vistula river, where, even after 21 days from the initial accident, some parameters remained altered.

Polymorphic microsatellite markers demonstrate hybridization and interspecific gene flow between lumbricid earthworm species, Eisenia andrei and E. fetida.

The lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) have been used as model organisms for studies on hybridization. Previously they have been identified by species specific sequences of the mitochondrial COI gene of maternal origin ('a' or 'f') and the nuclear 28S gene of maternal/paternal origin ('A' or 'F'). In experimental crosses, these hermaphroditic species produce progeny of genotypes Ea (aAA), Ef (fFF) and hybrids (aAF and fFA) originating by self-fertilization or cross-fertilization. To facilitate studies on new aspects of the breeding biology and hybridization of earthworms, polymorphic microsatellite markers were developed based on 12 Ea and 12 Ef specimens and validated on DNA samples extracted from 24 genotyped specimens (aAA, fFF, aAF and fFA) from three laboratory-raised families and 10 of them were applied in the present study. The results indicate that microsatellite markers are valuable tools for tracking interspecific gene flow between these species.

The effects of exogenous application of melatonin on the degradation of polycyclic aromatic hydrocarbons in the rhizosphere of Festuca.

The study aimed to assess the effects of melatonin, a plant growth regulator, on the degradation of phenanthrene (Phe) and pyrene (Py), in the rhizosphere of the Festuca grass. The experiments were divided into the following groups: 1) soil contaminated with Phe and Py, without the Festuca, 2) contaminated soil + Festuca, 3-5), contaminated soil + Festuca + the application of melatonin in three separate doses: 10, 50, or 100 µM. After 90 days, the effects of melatonin supplementation on the degradation of polycyclic aromatic hydrocarbons (PAHs) were analyzed by evaluating the rate of PAHs degradation, the expression of genes encoding salicylaldehyde dehydrogenase (SDH) and glutathione peroxidase (GPX) enzymes in Pseudomonas putida, as well as by measuring the total activity of dehydrogenase and peroxidase enzymes. Our results have shown that in soil contaminated by 300 mg kg-1 PAHs, application of melatonin (10, 50, 100 µM), resulted in the following increase in the dehydrogenase and peroxidase activity in all three applied doses (19% and 5.7%), (45.3% and 34.3%), (40.9% and 14.3%), respectively in comparison to the control group. The experiment showed that soil supplementation with melatonin at 50 µM, resulted in the highest removal rate of PAHs. According to our results, melatonin demonstrated a potentially favorable role in enhancing plant biomass, as well as an increase in soil bacterial population, and the activity of antioxidative enzymes in P. putida, causing all tested parameters of the soil and the expression of desired genes to be advantageously altered for the degradation of the chosen PAHs.

Species-specific Cd-detoxification mechanisms in lumbricid earthworms Eisenia andrei, Eisenia fetida and their hybrids.

Hermaphroditic lumbricid Eisenia sp. earthworms are ubiquitous and highly resistant to a variety of environmental stressors, including heavy metals. Among the progeny of laboratory mated inter-specific pairs of Eisenia fetida (Ea) and Eisenia andrei (Ef) there are fertile Ha hybrids derived from Ea ova fertilized by Ef spermatozoa and very rare sterile Hf hybrids from Ef ova fertilized by Ea spermatozoa. The aim of the first part of the experiment was to compare the life traits and whole body accumulation of cadmium in adult earthworms from genetically defined Ea, Ef and their hybrids (Ha) exposed for four weeks to commercial soil either unpolluted (control) or cadmium-spiked leading to moderate (M) or high (H) soil pollution (M = 425 and H = 835 mg kg-1 dry soil weight). Such exposure impaired cocoon production but not affected earthworm viability despite the massive Cd bioaccumulation in the whole earthworm bodies reaching at M and H groups 316-454, 203-338, 114-253, and 377-309 mg kg-1 dry body weights of Ea, Ef1, Ef2, and Ha, respectively, surprisingly reaching maximum accumulation quantities in hybrids. The second part of the experiment aimed to investigate cadmium-related defense mechanisms at transcriptomic level in coelomocytes non-invasively extruded from coelomic cavities of the new sets of Ea, Ef, Ha, and Hf earthworms exposed to Cd in microcosms for 0 days (control), 2 days, and 7 days (M = 425 mg kg-1). Expression level of stress-induced Cd-metallothionein (mt) and superoxide dismutase (sod) were gradually up-regulated, while the immune-connected lysenin (lys) was rapidly down-regulated; the expression of glutathione S-transferase (gst) and phytochelatin synthase (pcs) remained unaffected. Mt and sod gene up-regulation and lys gene down-regulation were especially pronounced in Ea-derived hybrids. In sum, capacity of cadmium bioaccumulation and detoxification mechanisms is more efficient in interspecific hybrids than in the pure Ea and Ef species.

Modeling and optimizing the removal of cadmium by Sinapis alba L. from contaminated soil via Response Surface Methodology and Artificial Neural Networks during assisted phytoremediation with sewage sludge.

The study was aimed to model and optimize the removal of cadmium from contaminated post-industrial soil via Sinapis alba L. by comparing two modeling approaches: Response Surface Methodology (RSM) and Artificial Neural Networks (ANN). The experimental design was done using the Box-Behnken Design method. In the RSM model, the quadratic model was shown to predict the closest results in comparison to our experimental data. For ANN approach, a two-layer Feed-Forward Back-Propagation Neural Network model was designed. The results showed that sewage sludge supplementation increased the efficiency of the Sinapis alba plant in removing Cd from the soil. After 28 days of exposure, the removal rate varied from 10.96% without any supplementation to 65.9% after supplementation with the highest possible (law allowed) dose of sewage sludge. The comparison proved that the prediction capability of the ANN model was much higher than that of the RSM model (adjusted R-square: 0.98, standard error of the Cd prediction removal: 0.85 ± 0.02). Thus, the ANN model could be used for the prediction of heavy metal removal during assisted phytoremediation with sewage sludge. Moreover, such approach could also be used to determinate the dose of sewage sludge that will ensure highest process efficiency.

Effects of sewage sludge supplementation on heavy metal accumulation and the expression of ABC transporters in Sinapis alba L. during assisted phytoremediation of contaminated sites.

ATP binding cassette (ABC) transporters, types C, G, and B were monitored via qPCR in order to investigate the influence of heavy metal (HM) contamination of post-industrial and post-agricultural soils and the effects of its supplementation with sewage sludge, on Sinapis alba plants. Five house-keeping genes were selected and validated to ensure the best reference points. The relative expression of ABC types C and G genes was profoundly affected by experimental conditions and included their upregulation after plants exposure to heavy metals and downregulation after supplementation with sewage sludge. However, ABC type C was more responsive then type G. The experimental conditions altered the expression of ABC type C gene faster than ABC type G and thus, the expression of ABC type C can therefore potentially be used as a bioindicator during assisted phytoremediation of degraded sites. In clean soil, supplementation with sewage sludge with a slight content of heavy metals still caused an upregulation in the expression of ABC types C and G, which showed that proper toxicity assessments are necessary to ensure safe application of sewage sludge into soils. Results showed that the analysed genes take a significant part in plants metal detoxification and that their expression is regulated at transcriptional level after exposure to soil contaminated with heavy metals by both, industrial activities and by sewage sludge supplementation. Thus, their expression can potentially be used as an early-warning biomarker when soil supplementation with sewage sludge is incorporated into the soil-management process.

Modelling assisted phytoremediation of soils contaminated with heavy metals - Main opportunities, limitations, decision making and future prospects.

The heavy metals (HMs) soils contamination is a growing concern since HMs are not biodegradable and can accumulate in all living organisms causing a threat to plants and animals, including humans. Phytoremediation is a cost-efficient technology that uses plants to remove, transform or detoxify contaminants. In recent years, phytoremediation is entering the stage of large-scale modelling via various mathematical models. Such models can be useful tools to further our understanding and predicting of the processes that influence the efficiency of phytoremediation and to precisely plan such actions on a large-scale. When dealing with extremely complicated and challenging variables like the interactions between the climate, soil and plants, modelling before starting an operation can significantly reduce the time and cost of such process by granting us an accurate prediction of possible outcomes. Research on the applicability of different modelling approaches is ongoing and presented work compares and discusses available models in order to point out their specific strengths and weaknesses in given scenarios. The main aim of this paper is to critically evaluate the main advantages and limitations of available models for large-scale phytoremediation including, among others, the Decision Support System (DSS), Response Surface Methodology (RSM), BALANS, PLANTIX and various regression models. Study compares their applicability and highlight existing gaps in current knowledge with a special reference to improving the efficiency of large-scale phytoremediation of sites contaminated with heavy-metals. The presented work can serve as a useful tool when choosing the most suitable model for the phytoremediation of contaminated sites.

Gene expression, DNA damage and other stress markers in Sinapis alba L. exposed to heavy metals with special reference to sewage sludge application on contaminated sites.

Bioindicators are promising tools used to detect the long-term effects of selected biosolids on plants development and should be implemented before large-scale supplementation of sewage sludge into the soil. The presented study shows the impact of sewage sludge application on metal-sensitive toxicity biological parameters (biomarkers) in Sinapis alba including: germination, root length, the activity of guaiacol peroxidase, the chlorophyll content, the level of DNA damage and the expression level of Ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and metallothionein (mt). We evaluated data from selected biomarkers in order to broaden our understanding of plants defense mechanisms against heavy metal contamination and the application of sewage sludge into soils. Overall, in contaminated soil after supplementation with both municipal sewage sludges, an increase in toxicity was noticed in DNA damage, mt and rbcl expression and total chlorophyll content. The supplementation of both soils with municipal sewage sludge caused a two-time induction in the mt expression. Moreover, clean soil supplemented with sewage sludge caused an increase in DNA damage shown as the tail moment from approximately 12 µm on control to 40 µm after supplementation. Even if those biosolids increased the initial germination, roots length, and biomass in comparison to the unamended soil, the toxicity was evidenced with other stress markers. Results showed, that in order to accurately assess the influence of sewage sludge application on plants the use of several specific biomarkers is required for safe land restoration. The conducted study also confirmed, both under biochemical and genotoxic tests, that iron enrichment for biosolids or contaminated soil can significantly reduce the bioavailability and toxicity of other metals.

Bioaccumulation, antioxidative response, and metallothionein expression in Lupinus luteus L. exposed to heavy metals and silver nanoparticles.

Yellow-lupin (Lupinus luteus L.) was grown on soils contaminated with heavy metals during two parallel studies. In the first one, the soil was contaminated by industrial activities whereas, in the second one, the soil was artificially contaminated with a single metal including Cd, Pb, Zn, Ni (in nitrate form), and Ag (in nitrate and nanoparticles form). The study was performed to assess a plant's response to contamination including its antioxidative response and molecular mechanisms involved in metal detoxification through the expression level of metallothioneins (MTs). Overall, the study provided insights into identification and validation of housekeeping genes (HKG) in L. luteus under exposure to metal stress and showed the effects of selected heavy metals and silver nanoparticles on the expression of metallothioneins, the activity of guaiacol peroxidase (GPX) and bioaccumulation of metals in leaves of L. luteus. As such, HKG validation using BestKeeper, NormFinder, and geNorm software allowed for the selection of four most stable reference genes in a context metal contamination for the selected plant. Moreover, a significant increase in the expression levels of MT was observed in plants grown under heavy metal stress and none on plants grown on 25 mg kg-1 of silver nanoparticles. Also, the GPX activity and MT expression showed statistically significant changes between different conditions and doses which means that they can be used as highly sensitive stress markers for planning the phytoremediation process on a large scale.