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This study incorporated hydrogeochemical facies, the entropy-weighted water quality index (EWQI), multivariate statistics, and probabilistic human exposure assessment to investigate hydrogeochemistry, analyze groundwater quality, and estimate potential risks to human health in a lithium-rich ore area (Jadar River basin, Serbia). The findings designated the Ca·Mg-HCO 3 hydrogeochemical type as the predominant type of groundwater, in which rock weathering and evaporation control the major ion chemistry. Due to the weathering of a lithium-rich mineral (Jadarite), the lithium content in the groundwater was very high, up to 567 mg/L, with a median value of 4.3 mg/L. According to the calculated EWQI, 86.4% of the samples belong to poor and extremely poor quality water for drinking. Geospatial mapping of the studied area uncovered several hotspots of severely contaminated groundwater. The risk assessment results show that groundwater contaminants pose significant non-carcinogenic and carcinogenic human health risks to residents, with most samples exceeding the allowable limits for the hazard index (HI) and the incremental lifetime cancer risk (ILCR). The ingestion exposure pathway has been identified as a critical contaminant route. Monte Carlo risk simulation made apparent that the likelihood of developing cancerous diseases is very high for both age groups. Sensitivity analysis highlighted ingestion rate and human body weight as the two most influential exposure factors on the variability of health risk assessment outcomes., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Historical changes of sediment characteristics and levels of inorganic and organic contaminants were studied in dated sediment cores from the Visovac Lake, situated in the Krka National Park, Croatia, to identify the main sources of anthropogenic pressures on this highly protected system. Depth distributions of lithogenic elements showed a steady decrease of terrigenous inputs due to the reduction in agricultural activities in the area, which was particularly pronounced during the 1991-1995 war in Croatia. Vertical and longitudinal distributions of Cd and Zn indicated that they are predominately of anthropogenic origin. The historical profiles of these toxic metals coincide well with the recorded production of metal industry in the upper reach of the Krka River with a sharp decrease reflecting the interruption by the war and slow recovery afterwards. By contrast, the recovery of the tourist industry in Krka NP after the war was accompanied by increasing contamination by elements characteristic of boat and car traffic (Sn, Cu, Pb) as well as oil pollution. The contamination with polycyclic aromatic hydrocarbons and polychlorinated biphenyls was only moderate. Although levels of metallic and organic contamination can be considered relatively low, the observed shift from industrial to tourism-related sources indicated that touristic activities should also be regarded as a possible threat for this vulnerable karst aquatic ecosystem., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Due to rapid urbanization and industrial growth, groundwater globally is continuously deteriorating, posing significant health risks to humans. This study employed a comprehensive methodology to analyze groundwater in the Western Banat Plain (Serbia). Using Piper and Gibbs plots, hydrogeochemistry was assessed, while the entropy-weighted water quality index (EWQI) was used to evaluate groundwater quality. Pollution sources were identified using positive matrix factorization (PMF) accompanied by Pearson correlation and hierarchical cluster analysis, while Monte Carlo simulation assessed health risks associated with groundwater consumption. Results showed that groundwater, mainly Ca-Mg-HCO 3 type, is mostly suitable for drinking. Geogenic pollution, agricultural activities, and sewage were major pollution sources. Consumption of contaminated groundwater poses serious non-carcinogenic and carcinogenic health risks. Additionally, arsenic from geogenic source was found to be the main health risks contributor, considering its worryingly elevated concentration, ranging up to 364 μg/L. These findings will be valuable for decision-makers and researchers in managing groundwater vulnerability. PRACTITIONER POINTS: Groundwater is severely contaminated with As in the northern part of the study area. The predominant hydrochemical type of groundwater in the area is Ca-Mg-HCO 3 . The PMF method apportioned three groundwater pollution sources. Monte Carlo identified rock dissolution as the primary health risk contributor. Health risks and mortality in the study area are positively correlated., (© 2024 Water Environment Federation.)

Considering the adaptability and responsiveness of microorganisms to environmental changes, their indicator potential is still not acknowledged in European directives. This comprehensive study examined the changes of microbial communities in sediments and a range of geochemical parameters from pristine and anthropogenically impacted coastal areas in the eastern Adriatic Sea. Various analytical methods found evidence of sediment contamination (high toxicity level, enrichments of metals, tributyltin) in certain areas, leading to the categorization of sediments based on the level of anthropogenic disturbance. Prokaryotes were identified as the most promising group of microbes for further research, with specific bacterial families (Rhodobacteraceae, Ectothiorhodospiraceae, Cyclobacteriaceae) and genera (Boseongicola, B2M28, Subgroup 23, Sva0485, Thiogranum) proposed as potential indicators of environmental status. Finally, predictive models were developed to identify key indicator variables for assessing anthropogenic impact in sediments. This research represents an essential step toward incorporating microbial communities into assessments of benthic environmental health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, we investigated the levels of macro, minor, and trace elements in roasted ground and instant coffees (n = 56). We assessed dietary mineral intake and health risks associated with potentially toxic elements (PTEs) using deterministic and probabilistic approaches. The limits of detection (LOD) ranged from 0.13 µg/kg for Be to 3.7 mg/kg for K, with corresponding limits of quantification (LOQ) at 0.43 µg/kg and 12.2 mg/kg. The recovery values (R%) ranged from 89 to 107%. The most abundant element was K, followed by Mg, Ca, and Na. Other elements followed this order: Fe > Mn > B > Cu > Sr > Zn > Al > Ba > Ni > Cr > Co > Se > Sn > Pb > Li > Ag > V > As > Cd > Hg > Be. Instant coffees generally exhibited higher K, Mg, and Na levels than ground-roasted coffees. Notably, Hg, Li, and Se were not detected in 34, 2, and 1 samples, respectively. Coffee samples were generally a good source of dietary elements such as Cu, Mn, Cr, and Se. The PTEs found in coffee products posed negligible risks to human health. The total target hazard quotient (TTHQ) remained below 1, and the incremental lifetime cancer risk (ILCR) did not exceed the threshold of 1 × 10 -6 . Nevertheless, coffee consumption contributed to Pb and As levels below 15% of the benchmark dose lower confidence limit (BMDL) values, and Sn, Hg, and Cd levels below 0.90% of the provisional tolerable weekly intake (PTWI)., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Leafy plants are commonly consumed as vegetables in India due to their high nutrient and vitamin content. This study, conducted in Ambagarh Chowki (India), investigated the accumulation potential of 52 elements (including Al, As, Ba, Be, Bi, Ca, Cd, Ce, Co, Cr, Cu, Dy, Er, Eu, Fe, Ga, Gd, Ge, Ho, K, La, Li, Lu, Mg, Mn, Mo, Na, Nb, Nd, Ni, P, Pb, Pr, Rb, Sb, Sc, Se, Sm, Sn, Sr, Tb, Te, Th, Ti, Tl, Tm, U, V, W, Y, Yb, and Zn) in seven leafy vegetable species, namely Amaranthus tricolor L., Corchorus olitorius L., Cordia myxa L., Hibiscus sabdariffa L., Ipomoea batatas (L.) Lam., Moringa oleifera Lam., and Spinacia oleracea L. Technique: Inductively coupled plasma mass spectrometry (ICP-MS) was employed for analysis. The maximum concentrations of elements such as Al, Ba, Be, Bi, Cd, Co, Cr, Fe, Ga, Ge, Li, Mn, Ni, Pb, Sb, Th, Tl, U, V, W, and REEs were observed in S. oleracea leaves, indicating their highest accumulation potential. In contrast, the maximum concentrations of As were found in H. sabdariffa leaves; Ca and Si in M. oleifera leaves; Mg, Sr, and Mo in A. tricolor leaves; and P, K, Cu, and Zn in C. myxa leaves, respectively. Twenty-one elements (Cr, Cd, Pb, Ni, Co, V, Cu, Zn, Fe, Mn, Th, Sb, Ba, Be, Li, Sr, Tl, U, Se, Sn, and REEs) exceeded permissible limits set by the WHO. The elevated hazard index values indicated significant non-carcinogenic effects. The sources of these elements could be attributed to a combination of geological factors and agricultural practices. This study highlights the need for further investigation into the potential health implications of consuming these vegetables in the aforementioned region., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A common evolutionary mechanism in biology to drive function is protein oligomerization. In prokaryotes, the symmetrical assembly of repeating protein units to form homomers is widespread, yet consideration in vitro of whether such assemblies have functional or mechanistic consequences is often overlooked. Dye-decolorizing peroxidases (DyPs) are one such example, where their dimeric α + β barrel units can form various oligomeric states, but the oligomer influence, if any, on mechanism and function has received little attention. In this work, we have explored the oligomeric state of three DyPs found in Streptomyces lividans, each with very different mechanistic behaviors in their reactions with hydrogen peroxide and organic substrates. Using analytical ultracentrifugation, we reveal that except for one of the A-type DyPs where only a single sedimenting species is detected, oligomer states ranging from homodimers to dodecamers are prevalent in solution. Using cryo-EM on preparations of the B-type DyP, we determined a 3.02 Å resolution structure of a hexamer assembly that corresponds to the dominant oligomeric state in solution as determined by analytical ultracentrifugation. Furthermore, cryo-EM data detected sub-populations of higher-order oligomers, with one of these formed by an arrangement of two B-type DyP hexamers to give a dodecamer assembly. Our solution and structural insights of these oligomer states provide a new framework to consider previous mechanistic studies of these DyP members and are discussed in terms of long-range electron transfer for substrate oxidation and in the "storage" of oxidizable equivalents on the heme until a two-electron donor is available., (© 2024 The Author(s). Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.)

Population growth, urbanization, industry, floods, and agriculture globally degrade groundwater in river plains, necessitating action for its quality assessment and management. Hence, a comprehensive methodology, including hydrogeochemical facies (Piper, Gibbs), irrigation indices (SAR, Wilcox), entropy-weighted water quality index (EWQI), positive matrix factorization (PMF), and Monte Carlo simulation of source-specific health risks was used in this study to analyze groundwater in the Morava river plain (Serbia). The results revealed a prevalent Ca-Mg-HCO3 groundwater type, influenced by water-rock interactions. Although groundwater was found suitable for irrigation, only 66.7 % of the samples were considered drinkable. Agricultural activities, natural processes, and municipal wastewater were identified as primary pollution sources. The incremental lifetime cancer risk (ILCR) and hazard index (HI) threshold exceedance for adults and children ranged from 8.5 % to 39 % of the samples, with arsenic identified as the most risk-contributing contaminant. These findings provide valuable insights for researchers studying groundwater vulnerability in river plains., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

This study investigated the occurrence and distribution of rare earth elements (REEs), including 14 lanthanoids, scandium (Sc), and yttrium (Y), in groundwater around a large coal-fired thermal power plant (TPP). The ICP-MS technique was used to analyze 16 REEs in groundwater samples collected from monitoring wells. REE concentrations ranged from 59.9 to 758 ng/L, with an average of 290 ng/L. The most abundant was Sc, followed by La, accounting for 54.2% and 21.4% of the total REE concentration, respectively. Geospatial analysis revealed the REE enrichment at several hotspots near the TPP. The highest REE concentrations were observed near the TPP and ash landfill, decreasing with the distance from the plant and the landfill. REE fractionation ratios and anomalies suggested the Light REE dominance, comprising over 78% of the total REEs. Correlation and principal component analyses indicated similar behavior and sources for most REEs. Health risk assessment found hazard indices (HI) of 1.36 × 10 -3 and 1.98 × 10 -3 for adults and children, respectively, which are far below the permissible limit (HI = 1). Likewise, incremental lifetime cancer risks (ILCR) were all below 1 × 10 -6 . Nevertheless, ongoing ash disposal and potential accumulation in the environment could elevate the REE exposure over time.

This paper investigates occurrence of metal(loid)s, and size-dependent changes in their concentration in recent marine sediments from coastal and open-sea environments in the eastern Adriatic. Size fractionation of sediments was performed after removal of organic matter (OM), and the individual fractions, comprising particles below 8 μm, 4 μm, 2 μm, 1 μm and 0.45 μm, were analysed using HR ICP-MS. The concentrations of most elements increased with decreasing particle size, as a result of accumulation of clay minerals and Fe and Mn (oxyhydr)oxides. A decrease in concentrations was observed for Ba, Sr, Ti and U, due to lowering of the carbonate content and presence in the coarse-grained and heavy mineral fraction. The highest element concentrations were determined in the fraction comprising particles below 1 μm. Occasionally, depending on the sedimentological environment and/or the element in question, the peak concentrations occurred in the <2 μm or <0.45 μm fraction. The lowest size-dependent enrichment was observed for elements associated with aluminosilicates (Al, Be, Cs, Co, Fe, K, Li, Rb). A different size-dependent behaviour of the elements was observed between deep-sea areas and shallow environments under greater coastal influence, mainly due to differences in sediment sorting, and between the northern and central vs. southern Adriatic due to the different catchment geologies. The Fe and Mn (oxyhydr)oxides, abundant in the deep-sea sediments, played an important role in the geochemical cycle of As, Cd, Co, Mo, Sb and V., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

A capacity to determine the provenance of high-value food products is of high scientific and economic interest. With the aim to develop a tool for geographical traceability of Croatian extra virgin olive oils (EVOO), multielement composition and 13 C/ 12 C isotope ratio in EVOO as well as the geochemistry of the associated soils were analysed in samples collected from three regions along the Croatian Adriatic coast. Soil geochemistry was shown to influence the transfer and elemental composition of EVOO. The most discriminating variables to distinguish EVOO from different regions were S, Mo, Rb, Mg, Pb, Mn, Sn, K, V and δ 13 C. The predictive models achieved high sensitivity and specificity, especially when carbon isotope composition was added. The results suggest that interregional geographical traceability of Croatian EVOO is possible based on matching their multielement composition with that of the soils in the provenance area., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In heme enzymes, such as members of the dye-decolorising peroxidase (DyP) family, the formation of the highly oxidising catalytic Fe(iv)-oxo intermediates following reaction with hydrogen peroxide can lead to free radical migration (hole hopping) from the heme to form cationic tyrosine and/or tryptophan radicals. These species are highly oxidising (∼1 V vs. NHE) and under certain circumstances can catalyse the oxidation of organic substrates. Factors that govern which specific tyrosine or tryptophan the free radical migrates to in heme enzymes are not well understood, although in the case of tyrosyl radical formation the nearby proximity of a proton acceptor is a recognised facilitating factor. By using an A-type member of the DyP family (DtpAa) as an exemplar, we combine protein engineering, X-ray crystallography, hole-hopping calculations, EPR spectroscopy and kinetic modelling to provide compelling new insights into the control of radical migration pathways following reaction of the heme with hydrogen peroxide. We demonstrate that the presence of a tryptophan/tyrosine dyad motif displaying a T-shaped orientation of aromatic rings on the proximal side of the heme dominates the radical migration landscape in wild-type DtpAa and continues to do so following the rational engineering into DtpAa of a previously identified radical migration pathway in an A-type homolog on the distal side of the heme. Only on disrupting the proximal dyad, through removal of an oxygen atom, does the radical migration pathway then switch to the engineered distal pathway to form the desired tyrosyl radical. Implications for protein design and biocatalysis are discussed., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

This study investigates the effects of ultrasound, in combination with chemical pretreatments, on the quality attributes (total phenolic and carotenoid content, antioxidant activity (2,2-Diphenyl-1-picrylhydrazyl assay (DPPH)), ferric-reducing ability (FRAP), CIE L* a* b* color, non-enzymatic browning, rehydration ratio, textural and morphological properties) of red pepper subjected to drying (hot air drying or freeze drying). The fractional factorial design was used to assess the impact of factors. The global Derringer desirability function was used to determine the optimal conditions for the best quality attributes of dried pepper. The drying method influenced total phenolic content, a* (redness), and initial rehydration ratio; pretreatment time significantly affected FRAP antiradical activity, a*, chroma and non-browning index, while pH-value had a significant effect on the texture of dried pepper. Non-enzymatic browning was reduced to 72.6%, while the DPPH antioxidant capacity of freeze-dried peppers was enhanced from 4.2% to 71.9%. Ultrasonic pretreatment led to changes in the pepper morphology, while potassium metabisulfite (KMS) was a more effective additive than citric acid., Competing Interests: The authors have declared no conflicts of interest for this article.

Pesticides are extensively used in the cultivation and postharvest protection of citrus fruits, therefore continuous monitoring and health risk assessments of their residues are required. This study aimed to investigate the occurrence of pesticide residues on citrus fruits and to evaluate the acute and chronic risk for adults and children. The risk ranking of twenty-three detected pesticides was carried out according to a matrix ranking scheme. Multiple residues were detected in 83% of 76 analyzed samples. In addition, 28% contained pesticides at or above maximum residue levels (MRLs). The most frequently detected pesticides were imazalil, azoxystrobin, and dimethomorph. According to the risk ranking method, imazalil was classified in the high-risk group, followed by prochloraz, chlorpyrifos, azinphos-methyl, tebufenpyrad, and fenpiroximate, which were considered to pose a medium risk. The majority of detected pesticides (74%) posed a low risk. The health risk assessment indicated that imazalil and thiabendazole contribute to acute (HQa) and chronic (HQc) dietary risk, respectively. The HQc was negligible for the general population, while the HQa of imazalil and thiabendazole exceeded the acceptable level in the worst-case scenario. Cumulative chronic/acute risk (HIc/HIa) assessment showed that chronic risk was acceptable in all samples for children and adults, while the acute risk was unacceptable in 5.3% of citrus fruits for adults and 26% of citrus fruits for children. Sensitivity analyses indicated that the ingestion rate and individual body weight were the most influential risk factors.

The remnants of historical industrial contamination can be detected in many aquatic ecosystems worldwide even at present time. Mrežnica is a river in Croatia that has been, for more than a hundred years, continually exposed to effluents of various industries, which have, in modern time, mostly ceased to operate. Our aim was to establish the level of current contamination and pollution of the Mrežnica river-water and sediments. The study of river contamination at three sites (reference site; site nearby former cotton industry facility in Duga Resa - DRF; industrial zone of Karlovac town - KIZ) in three sampling campaigns (May 2020, April and September 2021) encompassed analyses of physico-chemical water parameters, screening of 369 pesticides, measurement of metal (loid) concentrations in the sediments, and in the dissolved and particulate phases of the river-water. The sediment pollution was assessed through the analyses of total bacteria abundance (by targeting 16S rRNA genes), and their associated metal resistance genes (cnrA, pbrT and czcD) and class 1 integrons (intl1). At the DRF site, industrial organic contaminants that can be traced to textile production were detected (dye and nylon components), as well as increased levels of some metals bound to suspended particulate matter and sediments. At the most downstream KIZ site, occasional high level of industrial herbicide neburon was measured in the river-water, and metal contamination of suspended particulate matter and sediments was evident. Although, based on the comparison with legislation and literature data, the level of contamination was rather mild, the effects on microbial communities were unquestionable, confirmed by increased abundance of the czcD gene at DRF site and the intI1 gene at both industrially impacted sites. Obtained results indicated long-term sediment retention of some industrial contaminants at the places of historical freshwater contamination, and, thus, the necessity for their monitoring even after the termination of contamination sources., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Controlling the reactivity of high-valent Fe(IV)-O catalytic intermediates, Compounds I and II, generated in heme enzymes upon reaction with dioxygen or hydrogen peroxide, is important for function. It has been hypothesized that the presence (wet) or absence (dry) of distal heme pocket water molecules can influence whether Compound I undergoes sequential one-electron additions or a concerted two-electron reduction. To test this hypothesis, we investigate the role of water in the heme distal pocket of a dye-decolorizing peroxidase utilizing a combination of serial femtosecond crystallography and rapid kinetic studies. In a dry distal heme site, Compound I reduction proceeds through a mechanism in which Compound II concentration is low. This reaction shows a strong deuterium isotope effect, indicating that reduction is coupled to proton uptake. The resulting protonated Compound II (Fe(IV)-OH) rapidly reduces to the ferric state, giving the appearance of a two-electron transfer process. In a wet site, reduction of Compound I is faster, has no deuterium effect, and yields highly populated Compound II, which is subsequently reduced to the ferric form. This work provides a definitive experimental test of the hypothesis advanced in the literature that relates sequential or concerted electron transfer to Compound I in wet or dry distal heme sites., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

The history of metal pollution in the semi-enclosed and human-influenced marine system of the Boka Kotorska Bay (Southern Adriatic) was studied considering geological composition of the surrounding catchment, the sedimentation rate and the mineralogical and early diagenetic processes in the recent sediments. The determination of background concentrations of metals, undertaken for the first time in this environment, proved to be particularly important for Ni and Cr, which are naturally enriched in the sediments of the southern Adriatic. The results showed widespread moderate contamination with Pb and Sn since the 1970s, while the upper layers of sediments near shipyards, marinas and urban areas were more contaminated with Sn, Cu, Pb, Zn, Cd, As, Sb and Mo. The transport of material through the narrow straits separating different parts of Boka Kotorska Bay resulted in a different geochemical composition of the smaller bays and a limited distribution of contaminated sediments from local sources., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The spatial and temporal variability of dissolved trace elements (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Li, Mn, Mo, Ni, Pb, Sb, Sr, Ti, U, V and Zn) in the upper and middle reaches of the Sava River during two campaigns (2007/2008 and 2017/2018) was examined. The study also included important tributaries and wastewater effluents. No crucial change in metal concentrations in the Sava River water between the two sampling periods was detected. Elements Co, Cr, Cu, Fe, Sb and Zn had higher concentrations at several sites in the first period, whereas Ba, Cd and Pb in the second period. The influence of wastewater discharges proved to be negligible. The highest levels of potentially toxic metals (Cd, Cr, Cu, Sb, V and Zn) were found downstream of the industrial town of Sisak. Concentration-discharge relationship of dissolved elements showed a chemostatic behaviour for As, Cr, Cu, Sr and V and chemodynamic behaviour for all other elements. The UNMIX model, used to identify and quantify metal sources, indicated that soil weathering was the main source of most elements (Al, As, Ba, Co, Cu, Fe, Li, Ni, Sb, Sr and V), with an anthropogenic contribution for Cd, Mo, Pb, Ti and Zn, while Mn and U were associated with colloid formation. The Sava River was defined as moderately polluted regarding metals, whose water quality meets environmental quality standards (EQS). These results may help to better understand the behaviour of dissolved metals in rivers under anthropogenic pressure and to control pollution in the Sava River Basin., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

The increased demand for technology-critical elements (TCEs) in emerging technologies brings about the need to determine their spatial distribution in the environment and establish regulatory guidelines. In this paper, concentrations of Li, Sc, Nb, W, Ga, Ge, and REY (rare earth elements, including yttrium) in soils collected from different parent materials in the upper catchment of the Sava River (Slovenia, Croatia) were analysed. Results of multivariate (principal component analysis) and univariate (threshold methods) statistical techniques were used to determine geochemical characteristics of studied soils to identify the background variations and to establish geochemical threshold values. The investigated area is characterized by great lithological diversity and substantial variation of TCE concentrations. Among methods for assessment of geochemical threshold, the TIF (Tukey inner fence) and 97.5th percentile delivered the most reasonable results. Some exceedances above the 97.5th percentile were natural in origin, caused by local geology. These findings can provide baseline data because little is known about TCE variation on different geological substrata., (© 2020. Springer Science+Business Media, LLC, part of Springer Nature.)

The occurrence and vertical distribution of ten technology critical elements (TCEs) (Li, Nb, Sc, Ga, Y, La, Sb, Ge, Te, and W) were studied in sediment cores collected from remote freshwater and marine lakes (Plitvice, Visovac and Mir Lakes) in three protected areas of Croatia. These environmental archives were used to assess natural TCE levels in lake sediments and temporal trends in historical anthropogenic atmospheric deposition. TCE was determined after complete sediment digestion using high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS). The measured TCE concentrations spanned a wide range, which can be attributed to the varying input of terrigenous material into the studied lake systems. All obtained TCE concentrations were close to natural conditions and therefore could be used as a reference for other equivalent sediment systems in the coming years. The evaluation of anthropogenic influence on TCE concentrations showed a slight anthropogenic enrichment with Sb and Te in the upper sediment layers of some lakes (Plitvice and Mir Lakes), indicating a widespread atmospheric deposition, which, however, cannot be related to the recent increase in the use of TCE in modern technology., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Structure determination of proteins and enzymes by X-ray crystallography remains the most widely used approach to complement functional and mechanistic studies. Capturing the structures of intact redox states in metalloenzymes is critical for assigning the chemistry carried out by the metal in the catalytic cycle. Unfortunately, X-rays interact with protein crystals to generate solvated photoelectrons that can reduce redox active metals and hence change the coordination geometry and the coupled protein structure. Approaches to mitigate such site-specific radiation damage continue to be developed, but nevertheless application of such approaches to metalloenzymes in combination with mechanistic studies are often overlooked. In this review, we summarize our recent structural and kinetic studies on a set of three heme peroxidases found in the bacterium Streptomyces lividans that each belong to the dye decolourizing peroxidase (DyP) superfamily. Kinetically, each of these DyPs has a distinct reactivity with hydrogen peroxide. Through a combination of low dose synchrotron X-ray crystallography and zero dose serial femtosecond X-ray crystallography using an X-ray free electron laser (XFEL), high-resolution structures with unambiguous redox state assignment of the ferric and ferryl (Fe IV  = O) heme species have been obtained. Experiments using stopped-flow kinetics, solvent-isotope exchange and site-directed mutagenesis with this set of redox state validated DyP structures have provided the first comprehensive kinetic and structural framework for how DyPs can modulate their distal heme pocket Asp/Arg dyad to use either the Asp or the Arg to facilitate proton transfer and rate enhancement of peroxide heterolysis., (© 2021. The Author(s).)

Both O 2 and H 2 O 2 can oxidize iron at the ferroxidase center (FC) of Escherichia coli bacterioferritin (EcBfr) but mechanistic details of the two reactions need clarification. UV/Vis, EPR, and Mössbauer spectroscopies have been used to follow the reactions when apo-EcBfr, pre-loaded anaerobically with Fe 2+ , was exposed to O 2 or H 2 O 2 . We show that O 2 binds di-Fe 2+ FC reversibly, two Fe 2+ ions are oxidized in concert and a H 2 O 2 molecule is formed and released to the solution. This peroxide molecule further oxidizes another di-Fe 2+ FC, at a rate circa 1000 faster than O 2 , ensuring an overall 1:4 stoichiometry of iron oxidation by O 2 . Initially formed Fe 3+ can further react with H 2 O 2 (producing protein bound radicals) but relaxes within seconds to an H 2 O 2 -unreactive di-Fe 3+ form. The data obtained suggest that the primary role of EcBfr in vivo may be to detoxify H 2 O 2 rather than sequester iron., (© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Both O 2 and H 2 O 2 can oxidize iron at the ferroxidase center (FC) of Escherichia coli bacterioferritin (EcBfr) but mechanistic details of the two reactions need clarification. UV/Vis, EPR, and Mössbauer spectroscopies have been used to follow the reactions when apo-EcBfr, pre-loaded anaerobically with Fe 2+ , was exposed to O 2 or H 2 O 2 . We show that O 2 binds di-Fe 2+ FC reversibly, two Fe 2+ ions are oxidized in concert and a H 2 O 2 molecule is formed and released to the solution. This peroxide molecule further oxidizes another di-Fe 2+ FC, at a rate circa 1000 faster than O 2 , ensuring an overall 1:4 stoichiometry of iron oxidation by O 2 . Initially formed Fe 3+ can further react with H 2 O 2 (producing protein bound radicals) but relaxes within seconds to an H 2 O 2 -unreactive di-Fe 3+ form. The data obtained suggest that the primary role of EcBfr in vivo may be to detoxify H 2 O 2 rather than sequester iron., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Angewandte Chemie published by Wiley-VCH GmbH.)

Obtaining structures of intact redox states of metal centers derived from zero dose X-ray crystallography can advance our mechanistic understanding of metalloenzymes. In dye-decolorising heme peroxidases (DyPs), controversy exists regarding the mechanistic role of the distal heme residues aspartate and arginine in the heterolysis of peroxide to form the catalytic intermediate compound I (Fe IV =O and a porphyrin cation radical). Using serial femtosecond X-ray crystallography (SFX), we have determined the pristine structures of the Fe III and Fe IV =O redox states of a B-type DyP. These structures reveal a water-free distal heme site that, together with the presence of an asparagine, imply the use of the distal arginine as a catalytic base. A combination of mutagenesis and kinetic studies corroborate such a role. Our SFX approach thus provides unique insight into how the distal heme site of DyPs can be tuned to select aspartate or arginine for the rate enhancement of peroxide heterolysis., (© 2020 The Authors. Published by Wiley-VCH GmbH.)

Dye decolourising peroxidases (DyPs) are oxidative haem containing enzymes that can oxidise organic substrates by first reacting with hydrogen peroxide. Herein, we have focused on two DyP homologs, DtpAa and DtpA, from the soil-dwelling bacterium Streptomyces lividans. By using X-ray crystallography, stopped-flow kinetics, deuterium kinetic isotope studies and EPR spectroscopy, we show that both DyPs react with peroxide to form compound I (a Fe IV [double bond, length as m-dash]O species and a porphyrin π-cation radical), via a common mechanism, but the reactivity and rate limits that define the mechanism are markedly different between the two homologs (DtpA forms compound I rapidly, no kinetic isotope effect; DtpAa 100-fold slower compound I formation and a distinct kinetic isotope effect). By determining the validated ferric X-ray structure of DtpAa and comparing it with the ferric DtpA structure, we attribute the kinetic differences to a subtle structural repositioning of the distal haem pocket Asp side chain. Through site-directed mutagenesis we show the acid-base catalyst responsible for proton-transfer to form compound I comprises a combination of a water molecule and the distal Asp. Compound I formation in the wild-type enzymes as well as their distal Asp variants is pH dependent, sharing a common ionisation equilibrium with an apparent pK a of ∼4.5-5.0. We attribute this pK a to the deprotonation/protonation of the haem bound H 2 O 2 . Our studies therefore reveal a mechanism for compound I formation in which the rate limit may be shifted from peroxide binding to proton-transfer controlled by the distal Asp position and the associated hydrogen-bonded water molecules.

Posttranslational modifications of immunoglobulins have been a topic of great interest and have been repeatedly reported as a major factor in disease pathology. Cost-effective, reproducible, and high-throughput (HTP) isolation of immunoglobulins from human serum is vital for studying the changes in protein structure and the following understanding of disease development. Although there are many methods for the isolation of specific immunoglobulin classes, only a few of them are applicable for isolation of all subtypes and variants. Here, we present the development of a scheme for fast and simultaneous affinity purification of α (A), γ (G), and μ (M) immunoglobulins from human serum through affinity monolith chromatography. Affinity-based monolithic columns with immobilized protein A, G, or L were used for antibody isolation. Monolithic stationary phases have a high surface accessibility of binding sites, large flow-through channels, and can be operated at high flow rates, making them the ideal supports for HTP isolation of biopolymers. The presented method can be used for HTP screening of human serum in order to simultaneously isolate all three above-mentioned immunoglobulins and determine their concentration and changes in their glycosylation pattern as potential prognostic and diagnostic disease biomarkers., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Case studies of unusual traits can provide unique snapshots of the effects of modified systems. In this study, we report on an individual from a Serbian family with the ability to rapidly, accurately and voluntarily speak backwards. We consider psychological, neural and genetic correlates of this trait to identify specific relevant neural mechanisms and new molecular pathways for working memory and speech-related tasks. EEG data suggest that the effect of word reversal precedes semantic integration of visually presented backward-words, and that event-related potentials above the frontal lobe are affected by both word reversal and the maintenance of backward-words in working memory. fMRI revealed that the left fusiform gyrus may facilitate the production of backward-speech. Exome sequencing identified three novel coding variants of potential significance in the RIC3, RIPK1 and ZBED5 genes. Taken together, our data suggest that, in this individual, the ability to speak backwards is afforded by an extraordinary working memory capacity. We hypothesise that this is served by cholinergic projections from the basal forebrain to the frontal cortex and supported by visual semantic loops within the left fusiform gyrus and that these neural processes may be mediated by a genetic mutation in RIC3; a chaperone for nicotinic acetylcholine receptors.

Cerebellar involvement in cognitive functions has been revealed in numerous anatomical, clinical and neuroimaging studies and several hypotheses about potential the role of the cerebellum in higher level brain function have been established. The aim of this study was to show involvement of the cerebellum in simple cognitive tasks. For this matter, we contrasted two tasks from the same semantic domain with specific cognitive content and level of practice: counting forward and counting backward. Twelve volunteers participated in this fMRI study and they were asked to perform both tasks within the same number range (1 to 30 and vice versa). Results showed greater activation in the right cerebellum for the task of counting forward than for counting backward, while for counting backward greater activation was found in prefrontal cortex, supplementary motor area, and anterior cingulate of both hemispheres. Our results correlate with already established hypotheses about cerebellar role in precise and smooth control, not only in well-trained motor but in well trained cognitive tasks as well.

This study reports the synthesis of polymeric matrices based on N-isopropylacrylamide and itaconic acid and its application for immobilization of lipase from Candida rugosa. The lipase was immobilized by entrapment method. Free and immobilized lipase activities, pH and temperature optima, and storage stability were investigated. The optimum temperature for free and entrapped lipase was found to be 40 and 45 °C, while the optimum pH was observed at pH 7 and 8, respectively. Both hydrolytic activity in an aqueous medium and esterolytic activity in an organic medium have been evaluated. Maximum reaction rate (V max) and Michaelis-Menten constants (K m ) were also determined for immobilized lipase. Storage stability of lipase was increased as a result of immobilization process. Furthermore, the operational stability and reusability of the immobilized lipase in esterification reaction have been studied, and it was observed that after 10 cycles, the residual activity for entrapped lipase was as high as 50%, implying that the developed hydrogel and immobilized system could provide a promising solution for the flavor ester synthesis at the industrial scale.

Number words could be considered as a part of general verbal lexicon, although clinical and scientific data suggest that there are specific brain areas that are involved in specific aspects of number and language processing. In this fMRI study we examine the brain response to two number words generation tasks that should be closely related to the process of language generation: counting forward and counting backward. Our results imply that both investigated tasks strongly rely on brain structures usually related to language generation processes with a distinct activation of the prefronatal area for both counting tasks. Furthermore, we found differences in brain response among the investigated tasks regarding activation in the posterior part of the superior temporal gyrus and parietal cortex for the task of counting backward. Those findings suggest that the tasks of counting rely on established language-related brain areas with specific differences depending on cognitive content of the specified task.

Introduction: Foetal tumours are relatively rare; prenatal diagnosis enables additional diagnostics, and thus the decision on the continuation of pregnancy and planning of delivery. The paper presents prenatal ultrasound diagnostics of foetal head tumour with additionally analyzed magnetic resonance imaging (MRI)., Case Outline: On ultrasound imaging, in a 27-year-old nullipara, a tumour of the foetal head was suspected at the 22nd gestational week. By consultative ultrasound, a 45 x 41 x 50 mm echogenic soft tissue tumour with hypoechogenic areas was confirmed, which originated from the left side of the skull spreading towards the neck on the left behind the ear. On colour Doppler no vascularisation was seen within the tumour but several vessels were seen along the border-line and on the surface. The scan showed no associated abnormalities. Karyotype was 46, XY. MRI scan using triplanar SSFSE and HASTE sequences was done at 26 weeks and confirmed a 84 x 45 x 71 mm tumour originating from the scalp, spreading along the endocranium over the entire surface of the fronto-temporal sqama and left parietal bone. The tumour did not disturb the integrity of the internal tabula, and did not spread intracranially. Based on these findings, the diagnosis of haemangioma was made. At 38 weeks gestation, a 3810 g male infant was delivered by Caesarean section. A bluish, well-defined, non-pulsatile tumour arising from the left frontoparietal region was seen, thus confirming the diagnosis of haemangioma., Conclusion: MRI improved delineation of the tumour facilitating better planning of postnatal management and mode of delivery, with the information on the postnatal course and prognosis. In the prenatal period, MRI following an ultrasound diagnosed foetal tumour, is an additional part of diagnostic examinations, and is not contraindicated during pregnancy.

Tympanometric examination of the air pressure in the middle ear and of the mobility of the tympano-ossicular system was carried out on 82 otologically "healthy" children. Pathologic tympanogram was found in 25.00 per cent of examined ears, i.e. in 10.37 per cent of ears with tympanogram type B and in 14.63 per cent of ears with tympanogram type C. The tonal audiometric results in children with pathologic tympanogram revealed the reduction of conductive hearing from 21 to 30 dB. In patients with tympanogram type B thanks to myringotomy a dense mucos and serous secretion was found in the middle ear of 88.24 per cent of children. In the authors' opinion tympanometry is a very advantageous method in early detection of diseases of the middle ear and deafness in children, especially of the forms with poor symptomatology and evolutive processes. The authors plead for the routine use of screening tympanometry in otologic practice, which should prevent conductive deafness in children and improve both therapeutic and social rehabilitation of children with damaged conductive hearing.