Lithium and strontium accumulation in native and invasive plants of the Sava River: Implications for bioindication and phytoremediation.

The objective of this study was to investigate the potential of native and invasive plant species for the uptake and accumulation of lithium (Li) and strontium (Sr) along the Sava River, focusing on their bioindication and phytoremediation capabilities. Sampling was carried out in riparian zones exposed to different pollution sources in Slovenia, Croatia, and Serbia. Plant samples of native (Salix alba, Populus alba, Populus nigra, Ulmus glabra, Juglans regia) and invasive (Amorpha fruticosa, Reynoutria japonica, Solidago canadensis, Impatiens glandulifera) species were collected. The content of Li and Sr was analyzed in the soils, roots, and leaves of the selected plants, as well as physical and chemical soil properties. Both Li and Sr content in the soils increased from the source to the mouth of the Sava River. The native species showed significant potential for Li and Sr accumulation based on the metal accumulation index. The highest Sr accumulation was measured in the leaves of Salix alba and the roots of Juglans regia, while the highest Li accumulation was measured in Ulmus glabra. Native species, especially Salix alba, proved to be better bioindicators of Li and Sr. Invasive species, especially Amorpha fruticosa and Impatiens glandulifera, showed a remarkable ability to translocate Sr and Li, respectively, to leaves. These results provide valuable insight into the suitability of plants for biomonitoring soil contamination and potential applications in phytoremediation strategies. In summary, the study shows the importance of native species in the context of the accumulation and bioindication of soil pollution.

Phytoremediation potential of invasive plant species for potentially toxic elements along the Sava River upstream.

Invasive plant species (IPS) have many characteristics that are necessary for successful phytoremediation and the accumulation of large amounts of potentially toxic elements (PTEs). The most common IPS from the source of the Sava River are Reynoutria japonica, Solidago canadensis and Impatiens glandulifera. Considering that the riparian soils of the Sava River are classified as moderately polluted, this study investigated their enrichment with PTEs (As, B, Cd, Cr, Cu, Ni, Pb, Zn) and the potential for accumulation of these elements in roots and leaves of the most common IPS. The soil and plant samples were prepared using the wet digestion method in CEM Mars 6 microwave oven. The content of PTEs in soil and plant samples was determined by ICP-OES. The results showed a moderate to very high enrichment of Cu in the soils. Impatiens glandulifera has the highest uptake capacity in leaves and phytoextraction ability of B, Cd, Cu and Zn. Reynoutria japonica has some potential for phytoextraction of Pb, Zn and especially B, while Solidago canadensis has potential for phytoextraction of B and Zn, while excluding Cr and Cu. The analyzes have shown that the studied species are not suitable for bioindication of PTEs in the riparian soils of the Sava River. Considering the enrichment of soils with Cu, Impatiens glandulifera is the most suitable species for phytoremediation of these elements among the studied species.

Green solution to riparian pollution: Populus alba L. potential for phytoremediation and bioindication of PTEs along the Sava river.

This study addresses the potential of Populus alba L. for bioindication and phytoremediation of the contaminated lower part of the Sava River. The main objectives are to assess soil contamination with potentially toxic elements (PTEs: As, B, Cd, Cr, Cu, Li, Ni, Pb, Sr, and Zn), evaluate their availability, and assess the phytoremediation and bioindication potential of Populus alba. Quantification of the PTE contents was performed using inductively coupled plasma optical emission spectroscopy (ICP-OES), while bioindication and phytoremediation potential were evaluated using accumulation indices. The study revealed phytotoxic contents of Cr, Cu, Ni and Zn in the riparian soils of the lower Sava River. The percentage of available Cd was high, but due to its low total content, its phytotoxic potential is limited. According the metal accumulation index, Populus alba exhibits significant potential to accumulate the PTEs studied (with accumulated toxic contents of B, Cr, Li, Sr, and Zn). The ability of Populus alba to accumulate and bioindicate Cd, Cr, and Ni is promising, as is its ability to potentially remediate B, Cd, and Zn. Copper deficiency in leaves resulted in a reduction in photosynthetic performance, but without visible morphological symptoms. The reduced photosynthetic capacity serves as an adaptive strategy for this species in response to toxic levels of PTEs. Since Populus alba is widely distributed in European riparian forests, it is a good candidate to address soil contamination through phytoremediation and bioindication techniques.

Ethnobotanical and ethnomedicinal research into medicinal plants in the Mt Stara Planina region (south-eastern Serbia, Western Balkans).

BACKGROUND: Ethnobotanical research in Southeast Europe-one of the most important European hotspots for biocultural diversity-is significant for the acquisition of Traditional Ecological Knowledge related to plants as well as for encouraging the development of local environments. The current ethnobotanical research was conducted in the region of Mt Stara Planina (south-eastern Serbia), which is characterised by rich phytodiversity with a large number of endemic and relict plant species. The aim of the study was to document the diversity of uses of medicinal plants and of traditional knowledge on their therapeutic uses. METHODS: Ethnobotanical data was collected through both open and semi-structured interviews with locals. Fifty-one inhabitants were interviewed (26 men and 25 women), aged 30-91, and data was analysed by means of use reports, citation frequency, use values (UV), and the informant consensus factor (ICF). RESULTS: The study identified 136 vascular medicinal plant taxa and one lichen species belonging to 53 families and 116 genera. Lamiaceae (19), Rosaceae (18), and Asteraceae (17) had the highest species diversity. The plant parts most commonly used to make a variety of herbal preparations were the aerial parts (54 citations), leaves (35 citations), fruits (20 citations), flowers (18 citations), and roots (16 citations), while the most common forms of preparation were teas (60.78%), consumption of fresh tubers, leaves, roots, and fructus (6.86%), compresses (5.88%), juices (5.39%), decoctions (3.92%), 'travarica' brandy (3.92%), and syrups (2.45%). Of the recorded species, 102 were administered orally, 17 topically, and 18 both orally and topically. The plants with a maximum use value (UV = 1) were Allium sativum, Allium ursinum, Gentiana asclepiadea, Gentiana cruciata, Gentiana lutea, Hypericum perforatum, Thymus serpyllum and Urtica dioica. The highest ICF value (ICF = 0.95) was recorded in the categories of Skin and Blood, Blood Forming Organs, and Immune Mechanism. CONCLUSIONS: This study shows that medicinal plants in the research area are an extremely important natural resource for the local population as they are an important component of their health culture and provide a better standard of living.

Relationship between potentially toxic elements and macrophyte communities in the Sava river.

Freshwater ecosystems are at significant risk of contamination by potentially toxic elements (PTEs) due to their high inherent toxicity, their persistence in the environment and their tendency to bioaccumulate in sediments and living organisms. We investigated aquatic macrophyte communities and the concentrations of As, Cu, Cd, Cr, Pb, Zn, Ni and Fe in water and sediment samples to identify a pollution pattern along the Sava River and to investigate the potential impact of these PTEs on the diversity and structure of macrophyte communities. The study, which covered 945 km of the Sava River, showed a downstream increase in sediment concentrations of the analyzed elements. Both species richness and alpha diversity of macrophyte communities also generally increase downstream. Ordinary and partial Mantel tests indicate that macrophyte communities are significantly correlated with sediment chemistry, but only weakly correlated with water chemistry. In the lowland regions (downstream), beta diversity decreases successively, which can be attributed to an increasing similarity of environmental conditions at downstream sites. Species richness is relatively low at sites with low concentrations of Cr, Cd, Fe, and Cu in the sediment. However, species richness increases to a certain extent with increasing element concentrations; as element concentrations increase further, species richness decreases, probably as a result of increased toxicity. Some species that are generally more tolerant to high concentrations of PTEs are: Ceratophyllum demersum, Iris pseudacorus, Najas marina, Butomus umbellatus, Vallisneria spiralis, Potamogeton gramineus and Bolboschoenus maritimus maritimus. Potamogeton perfoliatus and the moss species Cinclidotus fontinaloides and Fontinalis antipyretica have narrow ecological amplitudes in relation to the concentrations of PTEs in the sediment.

Association of monocyte CCR2 expression with obesity and insulin resistance in postmenopausal women.

PURPOSE: Monocytes actively participate in inflammatory mechanisms that contribute to the development of adipose tissue dysfunction and atherogenesis. The aim of this study was to evaluate the association of monocyte CCR2 chemokine receptor expression and intracellular oxidative burst with the metabolic and inflammatory factors related to body weight. METHODS: The study was performed in 67 postmenopausal women with normal, overweight and obese body mass index. Monocyte CCR2 surface expression and intracellular oxidative burst activity (determined using 2', 7'-dichlorofluorescin diacetate) were analyzed by flow cytometry. Serum levels of HMW adoponectin, monocyte chemoattractant protein-1 (MCP-1), insulin, glucose, lipids and C-reactive protein were determined. RESULTS: Subjects with homeostasis model assessment-estimated insulin resistance (HOMA-IR) above the median had significantly higher proportion of CCR2+ monocytes and higher mean fluorescence intensity (MFI) of CCR2 and oxidative burst. The proportion of CCR2+ monocytes and CCR2 MFI were correlated with body weight, body mass index, fat mass, insulin and HOMA-IR. Oxidative burst also correlated with anthropometric measures, fat mass and expression of CCR2. No correlations were found between these markers of monocyte activation and HMW adiponectin or monocyte chemoattractant protein-1. The absolute number of monocytes was associated with insulin and this association remained significant after adjusting for C-reactive protein. In the multiple regression model the monocyte number was determined to be an independent predictor of insulin level. CONCLUSION: These results provide support for significant associations of monocyte number and markers involved in monocyte activation with obesity and insulin resistance.

Fine needle aspiration cytology of metastatic Merkel cell carcinoma.

Merkel cell carcinoma (MCC) is uncommon cutaneous malignant neuroendocrine tumour of the elderly people with rapidly growing skin nodules found frequently on sun-exposed areas of the body. MCC is often an aggressive tumour with high tendency for local recurrence, lymph node involvement and distant metastases. This paper reports a case of metastatic MCC diagnosed by fine needle aspiration cytology (FNAC), flow cytometric deoxiribonucleated acid (DNA) analysis, pathohistology and electron microscopy. The cytological features in aspirate (stained with Papenheim and Papanicolaou staining) included increased cellularity, discohesive groups of small-to-medium size malignant cells with uniform, round-to-oval nuclei with moulding effect, fine chromatin, multiple micronucleoli and scanty cytoplasm. DNA flow cytometric analysis of the aspirate showed unexpected results for clinically aggressive behaviour of this tumour (the patient died after 21 months), and revealed that tumour contained diploid peak with DNA index of 1.1. The proliferation was high with elevated S-phase fraction (21%). The cytological diagnosis of possible metastatic MCC was confirmed by histological one as well as by electron microscopy presented the pathognomonic features for this tumour: dense-core neurosecretory granules with diameter of 100-250 nm surrounded by whorls of intermediate filaments. MCC provides an enormous challenge for the morphologist because of a wide range of differential diagnosis and for the clinician because this tumour has a highly malignant potential for local recurrence, nodal and distant spread and very often is combined with other tumours. Therefore it is important to perform FNAC of different lesions in the same patient because it can distinguish MCC from the other tumours.

Monocytes in metabolic disorders--opportunities for flow cytometry contributions.

Chronic inflammation has arised as a major underlying cause of atherosclerosis, obesity and diabetes. It is mediated by cells of innate immune system like macrophages but also by their antecedents, circulating monocytes. Roles of monocyte subsets and different markers of monocyte activation in the context of metabolic disorders have been reviewed. Applying cell based approach through flow cytometry in this field has resulted with new understanding of pathophysiologic mechanisms. Possible implications of these insights in diagnosis, prognosis and revealing of therapeutic targets in metabolic disorders remain a challenge for future.

Flow cytometric analysis of deep-seated lymph nodes.

Flow cytometry (FC) immunophenotyping is an important tool in the evaluation of lymphadenopathy and is widely used in the diagnosis of non-Hodgkin's lymphomas (NHLs) on fine-needle aspirates of lymph nodes and extranodal sites. Because at least 80% of NHLs are of B-cell type, detection of immunoglobulin (Ig) light-chain-restriction is the most commonly used method for confirmation of monoclonality. The aim of our study was to evaluate usefulness of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for FC analysis from deep-seated lymph nodes and to compare results of FC clonality analysis to cytomorphologic diagnosis of sampled lymph nodes. For cytological diagnosis direct smears were made, selected slide was stained for rapid-on site evaluation procedure. Sixteen patients with suspected NHL of deep-seated lymph nodes obtained by EUS-FNA were submitted for FC clonality analysis using four-color multiparameter flow cytometry stained with kappa/lambda/CD19/CD45. Clonality analysis was performed on 11 samples. Monoclonality was demonstrated in seven of 11 cases cytologically diagnosed as NHL and four of 11 cases cytologically diagnosed as benign were polyclonal. Our results show that EUS-FNAC with FC is a sensitive and specific tool in the diagnosis of deep-seated B-NHL. Cytologic diagnosis combined with FC clonality analysis can be performed in majority of cases and may eliminate need for open biopsy in some cases.

The accuracy of fine needle aspiration cytology and flow cytometry in evaluation of nodal and extranodal sites in patients with suspicion of lymphoma.

Today lymphomas are defined according to a combination of morphology, immunophenotype, genetic features and clinical presentation, so beside the pure cytomorphologic analysis in diagnosis of lymphoma ancillary techniques such as cytochemistry, immunocytochemistry, molecular diagnosis and flow cytometry (FC) are often used. Our goal was to determinate how is information given by fine-needle aspiration cytology (FNAC) and FC correlated with pathohistologic diagnosis and to evaluate ability to diagnose and subclassify malignant lymphomas by FNAC and FC. This study is a retrospective chart review of patients with suspicion of lymphoma processed at University Hospital Dubrava in Zagreb. After analysis 50 patients fulfilled inclusion criteria for this study (FNAC diagnosis with or without FC and consecutive confirmatory pathohistological diagnosis). When analyzing accuracy of FNAC according to suspicion of lymphoma or NHL and differential diagnosis lymphoma sensitivity was 97.7%, specificity 85.7% and the diagnostic accuracy was 96%. When analyzing accuracy of FNAC according to the subclassification of lymphoma, sensitivity was 74.4%, specificity 85.7% and the diagnostic accuracy 76%. Combined FNAC and FC improved sensitivity, positive predictive value, negative predictive value and diagnostic accuracy. Sensitivity was 79.1% and the diagnostic accuracy 80%. We have shown that these methods can distinguish benign lymphadenopaties from lymphomas and also subclassify lymphomas and quickly provide clinicians with that information.