Chemical analysis of wetland plants to evaluate the bioaccumulation and metabolism of hexachlorocyclohexane (HCH).

Hexachlorocyclohexanes (HCH) belong to the banned pesticides with short-time production and use during the last century. However, the consequences of this short period are still present as persistent environmental contamination. This study represents the large lab-scale experiment focused on the HCH accumulation and metabolism in selected wetland plants (Juncus effuses, Typha latifolia, Phragmites australis) and trees (Alnus glutinosa) after the exposure to the technical mix of HCH isomers (t-HCH) or δ-HCH at three different concentration. During the three-month exposure, morphological (biomass, height, relative chlorophyll content) and physiological (photosynthetic measurements - photosynthetic rate, stomatal conductance, transpiration and dark transpiration) parameters were measured to assess the HCH effect on plant's growth. The results showed that all selected plant species supported HCH removal from the soil. The total removal efficiency was lower for the t-HCH than for δ-HCH exposure, and the best results were provided by Alnus glutinosa tree. Also, no isomer preference was observed in plants exposed to t-HCH. Most HCH remained accumulated in the root biomass, and mainly α-HCH and δ-HCH were transported to the above-ground parts due to their physicochemical properties. Simultaneously, HCH uptake and metabolization to chlorobenzenes (CB) and chlorophenols (CP) occur. Non-targeted analysis showed that CP could be conjugated to glucose and malonyl in plant tissue, and secondary plant metabolism is affected positively and negatively after exposure to t-HCH depending on plant species and chemical concentration. Luteolin, quercetin and quercetin-3-O-glucoside found common to all species showed quantitative changes due to HCH. Nevertheless, most morphological and physiological parameters were adversely affected without statistical significance. This large-scale study provides information on the fate of HCH in the soil-plant system, the suitability of selected plants and their adaptation to chemical stress for use in the phytoremediation process.

Assessing HCH isomer uptake in Alnus glutinosa: implications for phytoremediation and microbial response.

Although the pesticide hexachlorocyclohexane (HCH) and its isomers have long been banned, their presence in the environment is still reported worldwide. In this study, we investigated the bioaccumulation potential of α, ß, and δ hexachlorocyclohexane (HCH) isomers in black alder saplings (Alnus glutinosa) to assess their environmental impact. Each isomer, at a concentration of 50 mg/kg, was individually mixed with soil, and triplicate setups, including a control without HCH, were monitored for three months with access to water. Gas chromatography-mass spectrometry revealed the highest concentrations of HCH isomers in roots, decreasing towards branches and leaves, with δ-HCH exhibiting the highest uptake (roots-14.7 µg/g, trunk-7.2 µg/g, branches-1.53 µg/g, leaves-1.88 µg/g). Interestingly, α-HCH was detected in high concentrations in ß-HCH polluted soil. Phytohormone analysis indicated altered cytokinin, jasmonate, abscisate, and gibberellin levels in A. glutinosa in response to HCH contamination. In addition, amplicon 16S rRNA sequencing was used to study the rhizosphere and soil microbial community. While rhizosphere microbial populations were generally similar in all HCH isomer samples, Pseudomonas spp. decreased across all HCH-amended samples, and Tomentella dominated in ß-HCH and control rhizosphere samples but was lowest in δ-HCH samples.

Biodiversity in wetland+ system: a passive solution for HCH dump effluents.

The hexachlorocyclohexane isomers (HCH) are long-banned pesticides. Even though their use has been prohibited for decades, their presence in the environment is still reported worldwide. Wetland + is a registered trademark of the remedial treatment technology consisting of an aerobic sedimentary tank, a permeable reactive barrier, a biosorption system, and an aerobic wetland. This proven method combines a reductive treatment known from PRBs with the natural wetland self-cleaning processes. The average efficiency of the system is 96.8% for chlorobenzenes (ClB) and 81.7% for HCH, during the first 12 months of the system operation. The presence of the genes encoding enzymes involved in the degradation of the HCH compounds indicates that the removal of HCH and ClB occurs not only by chemical removal but also through aerobic and anaerobic combining biodegradation. Changes in abundance and the composition of the diatom community were found to be suitable indicators of the water quality and of the impact of the Wetland + operation on the water ecosystem. The system's annual operation exhibited a markedly higher number of diatom species in the closing profiles of the Ostrovský Creek, the Wetland + effluent recipient.

Influence of delta-hexachlorocyclohexane (δ-HCH) to Phytophthora ×alni resistant Alnus glutinosa genotypes - Evaluation of physiological parameters and remediation potential.

Hexachlorocyclohexanes (HCHs) are persistent organochlorine pesticides with the adverse effects on human health and the environment. The effect of delta-isomer of hexachlorocyclohexane (δ-HCH) on germination, growth parameters and physiological parameters was studied in different Alnus glutinosa (L.) Gaertn. progeny of resistant genotypes to pathogen Phytophthora ×alni. Two experiments were performed: a short-term experiment to determine the effect of δ-HCH on total germination (GT), germination energy (GE), speed of germination (SG), shoot length and biomass of seedlings, and a long-term experiment devoted to remediation aspects. In addition, changes in the hormonal system of alders were monitored in both cases. Significant differences were found between the treated and control group in most of the evaluated characteristics. Also, the content of studied phytohormones differs between groups. Furthermore, the obtained results indicate genetically determined variability in response to δ-HCH. Of the six tested, the Brezové and Turany progeny seem to be suitable candidates for phytoremediation because of the adaptation to stress conditions or high remediation efficiency. The rest of tested progeny seems to be unsuitable due to higher mortality, lower remediation efficiency and higher levels of stress hormones resulting in significant decrease in biomass and plant height. Moreover, results indicate the role of the plant as a remediation accelerator, probably through released exudates, and a positive effect on the soil microbiome as the presence of plants increased the remediation efficiency by 20.85 - 35.89%. The obtained research findings may be helpful in better understanding the processes involved in removing these pesticides from the soil. Further research should be focused on rhizosphere microbiome, mechanism of in-plant isomerization and metabolites identification.

Phytoremediation potential of Miscanthus sinensis And. in organochlorine pesticides contaminated soil amended by Tween 20 and Activated carbon.

The organochlorine pesticides (OCPs) have raised concerns about being persistent and toxic to the environment. Phytoremediation techniques show promise for the revitalization of polluted soils. The current study focused on optimizing the phytoremediation potential of Miscanthus sinensis And. (M. sinensis), second-generation energy crop, by exploring two soil amendments: Tween 20 and activated carbon (AC). The results showed that when M. sinensis grew in OCP-polluted soil without amendments to it, the wide range of compounds, i.e., α-HCH, ß-HCH, γ-HCH, 2.4-DDD, 4.4-DDE, 4.4-DDD, 4.4-DDT, aldrin, dieldrin, and endrin, was accumulated by the plant. The introduction of soil amendments improved the growth parameters of M. sinensis. The adding of Tween 20 enhanced the absorption and transmigration to aboveground biomass for some OCPs; i.e., for γ-HCH, the increase was by 1.2, for 4.4-DDE by 8.7 times; this effect was due to the reduction of the hydrophobicity which made pesticides more bioavailable for the plant. The adding of AC reduced OCPs absorption by plants, consequently, for γ-HCH by 2.1 times, 4.4-DDD by 20.5 times, 4.4-DDE by 1.4 times, 4.4-DDT by 8 times, α-HCH was not adsorbed at all, and decreased the translocation to the aboveground biomass: for 4.4-DDD by 31 times, 4.4-DDE by 2.8 times, and γ-HCH by 2 times; this effect was due to the decrease in the bioavailability of pesticides. Overall, the amendment of OCP-polluted soil by Tween 20 speeds the remediation process, and incorporation of AC permitted to produce the relatively clean biomass for energy.

Prevalence of Rickettsia species in ticks including identification of unknown species in two regions in Kazakhstan.

BACKGROUND: Over 60 years ago clinical patterns resembling tick-borne rickettsioses have been described for the first time in Kazakhstan. Since 1995 the incidence of clinical cases of tick-borne rickettsioses in humans seems to be rising but studies on epidemiological data regarding the occurring etiological agents, tick vector species, prevalence and distribution throughout Kazakhstan are still scarce to date. The aim of the study was molecular investigation of ticks for spotted-fever group rickettsiae in the endemic Kyzylorda region and the so far considered as non-endemic Almaty region. A total of 2341 ticks was collected in the two regions in Kazakhstan and sorted in 501 pools: Ixodes persulcatus (243); Dermacentor marginatus (129); Haemaphysalis punctata (104); Hyalomma asiaticum (17); Dermacentor reticulatus (3); and Rhipicephalus turanicus (5). Pools were tested for Rickettsia spp. using real-time PCR. For positive samples multilocus sequence typing (MLST) was performed. RESULTS: The calculated minimum infection rate (MIR) for rickettsiae in the investigated ticks in Almaty region varied between 0.4-15.1% and 12.6-22.7% in the Kyzylorda region. At least four different Rickettsia species were identified in the two selected regions of Kazakhstan. Two of these are already known to science: Rickettsia raoultii and R. slovaca, the latter being reported for the first time in Almaty region One new form, "Candidatus R. yenbekshikazakhensis", was described by MLST of six gene fragments in Almaty region and one new genotype, "genotype R. talgarensis" was detected using three gene fragments. CONCLUSIONS: Kazakh physicians should be aware of rickettsioses after tick bites in both regions studied. Both, R. raoultii and R. slovaca should be included in the diagnostics. The role for human diseases has further to be investigated for the newly described rickettsiae, "Candidatus R. yenbekshikazakhensis" and "Genotype R. talgarensis".

Alterations of oncogenes expression in NK cells in patients with cancer.

INTRODUCTION: C-kit/SCF signaling plays a key role in regulating NK cell homeostasis, maturation, proliferation, and cytotoxicity. C-kit-deficiency in NK cells results in significant reduction of their number, suggesting an imperative role for c-kit signaling in NK cell biology. We have recently showed that human NK cells express not only c-kit-receptor, but also both membrane-bound and soluble forms of c-kit ligand-Stem cell factor. The goal of this study was to characterize the c-kit/SCF autocrine loop in peripheral blood NK cells obtained from patients with cancer. METHODS: Using Smart Flare and qRT-PCR, we have characterized expression of c-kit and two forms of SCF in patients' NK cells and correlated these results with the expression of c-myc and STAT3. RESULTS: Our results demonstrated that the expression of proto-oncogenes c-myc and c-kit was significantly decreased in NK cells from all cancer patients. Expression of membrane-bound SCF in NK cells correlated with the presence of remote metastases. CONCLUSIONS: We suggest that the abnormal signaling and expression of c-kit/SCF, c-myc, and STAT3 in NK cells is responsible for the defect in their cytolytic activity in cancer and these defects at the gene expression level may be the cause rather than the result of tumor progression.

Transcriptional profiling of interleukin-2-primed human adipose derived mesenchymal stem cells revealed dramatic changes in stem cells response imposed by replicative senescence.

Inflammation is a double-edged sword with both detrimental and beneficial consequences. Understanding of the mechanisms of crosstalk between the inflammatory milieu and human adult mesenchymal stem cells is an important basis for clinical efforts. Here, we investigate changes in the transcriptional response of human adipose-derived stem cells to physiologically relevant levels of IL-2 (IL-2 priming) upon replicative senescence. Our data suggest that replicative senescence might dramatically impede human mesenchymal stem cell (MSC) function via global transcriptional deregulation in response to IL-2. We uncovered a novel senescence-associated transcriptional signature in human adipose-derived MSCs hADSCs after exposure to pro-inflammatory environment: significant enhancement of the expression of the genes encoding potent growth factors and cytokines with anti-inflammatory and migration-promoting properties, as well as genes encoding angiogenic and anti-apoptotic promoting factors, all of which could participate in the establishment of a unique microenvironment. We observed transcriptional up-regulation of critical components of the nitric oxide synthase pathway (iNOS) in hADSCs upon replicative senescence suggesting, that senescent stem cells can acquire metastasis-promoting properties via stem cell-mediated immunosuppression. Our study highlights the importance of age as a factor when designing cell-based or pharmacological therapies for older patients and predicts measurable biomarkers characteristic of an environment that is conducive to cancer cells invasiveness and metastasis.