Oxidative potential, environmentally persistent free radicals and reactive oxygen species of size-resolved ambient particles near highways.

Particulate matter (PM) is a group of atmospheric pollutants with an uncertain toxicity, particularly when collected near highways. This study examined the oxidative potential (OP) of, as well as the environmentally persistent free radicals (EPFRs) and reactive oxygen species (ROS) present in PM samples collected near highways in Xiamen, China. Our findings revealed that PM had a relatively high OP, ranging from 3.8 to 18.5 nmol/min/µg, surpassing values reported in previous research. The oxidative potential of the water-insoluble fraction (OPWIS), which accounted for 68% of the total oxidative potential (OPTotal), demonstrated rapid toxicity, whereas the oxidative potential of the water-soluble fraction (OPWS) displayed a steadier toxicity release pattern. The primary free radicals detected in PM were oxygen-centered. The measured concentration of EPFRs was 6.073 × 1014 spins/m3, which is lower than that reported in previous studies, possibly because of the high relative humidity of the road environment in Xiamen. We also investigated the interaction between PM and water near highways and observed the generation of R and OH radicals. Additionally, we analysed the sample composition and evaluated the contributions of the different components to OPTotal. Transition metals (Fe, Cu, and Zn) were identified as the major contributors, accounting for 33.2% of the OPTotal. The positive correlation observed between EPFRs and ROS suggests that EPFRs may be involved in ROS generation. The correlation analysis indicated that the oxidative potential measured using the DTT method (OPDTT) could serve as an indicator of ROS generation. Finally, based on the relationship between OPDTT, EPFRs, and ROS, we propose that reducing the emission of transition metals, particularly Fe, represents an effective control measure for mitigating PM toxicity near highways.

Current Innovations of Metal Hexacyanoferrates-Based Nanocomposites toward Electrochemical Sensing: Materials Selection and Synthesis Methods.

Mixed valence transition metal hexacyanoferrates (MeHCF)-Prussian blue and its analogs receive enormous research interest in the electrochemical sensing field. In recent years, conducting materials such as conducting polymer, carbon nanomaterial, and noble metals have been used to form nanocomposites with MeHCF. The scope of this review offers the reasons behind the preparation of various MeHCF based nanocomposite toward electrochemical detection. We primarily focus on the current progress of the development of MEHCF-based nanocomposites. The synthesis methods for these nanocomposites are also reviewed and discussed.

Detection and quantification of transition metal leaching in metal affinity chromatography with hydroxynaphthol blue.

The widespread use of immobilized metal-affinity chromatography (IMAC) for fast and efficient purification of recombinant proteins has brought potentially toxic transition elements into common laboratory usage. However, there are few studies on the leaching of metal from the affinity resin, such as nickel-nitrilotriacetic acid (Ni-NTA), with possible deleterious impact on the biological activity. This is of particular importance when reducing or chelating eluants stronger than imidazole are used. We present a detailed study of hydroxynaphthol blue (HNB) as an indicator of several divalent metal cations, but with emphasis on Ni2+, clarifying and correcting many errors and ambiguities in the older literature on this dye compound. The assay is simple and sensitive and many metals, notably Ni2+, Zn2+, Cu2+, Pb2+, Fe2+, Co2+, and Al3+, can be readily detected and quantified at concentrations down to 15-50 nM (1-5 ppb) at neutral pH and in most commonly used buffers using spectroscopic equipment available in typical biochemistry research labs. Using this method, we show that significant amounts of Ni2+ (up to 20 mM) are co-purified with a target protein (cytochrome bc1 complex) when histidine is used to elute from Ni-NTA resin.

Size-resolved particle oxidative potential in the office, laboratory, and home: Evidence for the importance of water-soluble transition metals.

Particulate matter (PM) oxidative potential (OP) is an emerging health metric, but studies examining the OP of indoor PM are rare. This paper focuses on the relationships between respiratory exposure to OP and PM water-soluble composition in indoor environments. Size-resolved PM samples were collected between November 2015 and June 2016 from an office, home (including bedroom, living room, and storeroom), and laboratory using a MOUDI sampler. Particles from each source were segregated into eleven size bins, and the water-soluble metal content and dithiothreitol (DTT) loss rate were measured in each PM extract. The water-soluble OP (OPws) of indoor PM was highest in the office and lowest in the home, varying by factors of up to 1.2; these variations were attributed to differences in occupation density, occupant activity, and ventilation. In addition, the particulate Cu, Mn, and Fe concentrations were closely correlated with OPws in indoor particles; the transition metals may have acted as catalysts during oxidation processes, inducing ·OH formation through the concomitant consumption of DTT. The OPws particle size distributions featured single modes with peaks between 0.18 and 3.2 µm across all indoor sites, reflecting the dominant contribution of PM3.2 to total PM levels and the enhanced oxidative activity of the PM3.2 compared to PM>3.2. Lung-deposition model calculations indicated that PM3.2 dominated the pulmonary deposition of the OPws (>75%) due to both the high levels of metals content and the high deposition efficiency in the alveolar region. Therefore, because OPws has been directly linked to various health effects, special attention should be given to PM3.2.

Summer-winter differences of PM2.5 toxicity to human alveolar epithelial cells (A549) and the roles of transition metals.

Atmospheric fine particulate matters (PM2.5) induce adverse human health effects through inhalation, and the harmful effects of PM2.5 are determined not only by its air concentrations, but also by the particle components varied temporally. To investigate seasonal differences of the aerosol toxicity effects including cell viability and membrane damage, cell oxidative stress and responses of inflammatory cytokines, the human lung epithelial cells (A549) were exposed to PM2.5 samples collected in both summer and winter by the in vitro toxicity bioassays. Toxicological results showed that, the PM2.5 led to the cell viability decrease, cell membrane injury, oxidative stress level increase and inflammatory responses in a dose-dependent manner. Temporally, the cytotoxicity of winter PM2.5 was higher than summer of this studied industrial area of Nanjing, China. According to the different contents of heavy metals accumulated in PM2.5, the transition metals such as Cu might be an important contributor to the aerosol cell toxicity.

Transition metals and trace elements in the retinal pigment epithelium and choroid: correlative ultrastructural and chemical analysis by analytical electron microscopy and nano-secondary ion mass spectrometry.

Understanding the localisation and abundance of structural elements, trace elements and especially transition metals like Cu and Zn in ocular tissue sections is important for physiology, and also for the characterisation of diseases related to oxidative stress like age-related macular degeneration. Transition metal abundances were investigated in an aged donor eye by nano-secondary ion mass spectrometry (nano-SIMS) elemental mapping using Cs+ and O- primary ions, respectively, and correlated to their respective mole fractions investigated by analytical electron microscopy (AEM). The ultrastructure of the tissue and the elemental composition of melanosomes of the choroid and RPE, and RPE lipofuscin and melanolipofuscin granules can adequately be investigated by nano-SIMS using the secondary ion maps. Melanosomes, 0.5-1 µm in size, yield sulphur maps and maps of stored metals like calcium, sodium and copper. Lipofuscin shows especially high phosphorus signals. Elements with mole fractions of about 0.1 at%, e.g. for P and Cu, as investigated by AEM before, can be validated using simultaneous SIMS maps with an estimated lateral resolution of 66 nm with typical acquisition times of 30 minutes for each area of interest. However, Zn (0.19 at%) was not detected by SIMS. Nano-SIMS imaging of CN-, PO2-, S-, Cu-, Ca+, Fe+ and Na+ ions provides excellent detection limits demonstrating the possibilities for chemical mapping with high-sensitivity trace element detection and reduced acquisition times. Quantification of nano-SIMS data was achieved by correlating mole fractions obtained by AEM to secondary ions per pixel obtained by nano-SIMS. Both methods yield the melanin type in melanosomes and trace metal storage.

Genetically encoded fluorescent sensors for measuring transition and heavy metals in biological systems.

Great progress has been made in expanding the repertoire of genetically encoded fluorescent sensors for monitoring intracellular transition metals (TMs). This powerful toolkit permits dynamic and non-invasive detection of TMs with high spatial-temporal resolution, which enables us to better understand the roles of TM homeostasis in both physiological and pathological settings. Here we summarize the recent development of genetically encoded fluorescent sensors for intracellular detection of TMs such as zinc and copper, as well as heavy metals including lead, cadmium, mercury, and arsenic.

Trace Element Analysis of Borrelia burgdorferi by Inductively Coupled Plasma-Sector Field Mass Spectrometry.

Transition metal ions play important structural, regulatory, and catalytic roles in all biological systems by serving as cofactors for proteins. Due to their relatively low levels in the cell compared to abundant metal ions such as potassium and magnesium, transition metals are often considered micronutrients and referred to as trace elements. Manganese (Mn), iron (Fe), copper (Cu), and zinc (Zn) are the most prevalent transition metals in the Lyme disease spirochete Borrelia burgdorferi. Here, we describe a method for the accurate measurement of these trace elements in B. burgdorferi utilizing inductively coupled plasma-sector field mass spectrometry (ICP-SFMS).

Transition and post-transition metals in exhaled breath condensate.

Water vapor in expired air, as well as dispersed non-volatile components, condense onto a cooler surface after exiting the respiratory tract. This exhaled breath condensate (EBC) provides a dilute sampling of the epithelial lining fluid. Accordingly, the collection of EBC imparts a capacity to provide biomarkers of injury preceding clinical disease. Concentrations of transition and post-transition metals in EBC are included among these endpoints. Iron and zinc are the metals with the highest concentration and are measurable in all EBC samples from healthy subjects; other metals are most frequently either at or below the level of detection in this group. Gender, age, and smoking can impact EBC metal concentrations in healthy subjects. EBC metal concentrations among patients diagnosed with particular lung diseases (e.g. asthma, chronic obstructive disease, and interstitial lung disease) have been of research interest but no definite pattern of involvement has been delineated. Studies of occupationally exposed workers confirm significant exposure to specific metals, but such EBC metal measurements frequently provide evidence redundant with environmental sampling. Measurements of metal concentrations in EBC remain a research tool into metal homeostasis in the respiratory tract and participation of metals in disease pathogenesis. The quantification of metal concentrations in EBC is currently not reliable for clinical use in either supporting or determining any diagnosis. Issues that must be addressed prior to the use of EBC metal measurements include the establishment of both standardized collection and measurement techniques.

Role of ammonium ion and transition metals in the formation of secondary organic aerosol and metallo-organic complex within fog processed ambient deliquescent submicron particles collected in central part of Indo-Gangetic Plain.

In this study we observed the role of ammonium ion (NH4+) and transition metals (Fe, Mn, Cr, and Cu) present in ambient submicron particles in stabilizing and enhancing the yield of water soluble organic carbon (WSOC). A good correlation of WSOC with transition metals and NH4+ was found (R2 = 0.87 and 0.71), respectively within foggy episode collected ambient PM1 (particles having aerodynamic diameter ≤1.0 µm) suggesting plausibleness of alternate oxidation (primarily various carbonyls into their respective organic acids, esters and other derivatives.) and aging mechanisms. Molar concentration of ammonium ion was observed to be exceeded over and above to require in neutralizing the sulphate and nitrate which further hints its role in the neutralization, stabilization and enhancement of subset of WSOC such as water soluble organic acids. Transition metals were further apportioned using enrichment factor analysis. The source of Fe, Mn, and Cr was found to be crustal and Cu was tagged to anthropogenic origin. This study also described the plausible role of significant predictors (Fe and Cu) in the secondary organic aerosol (SOA) formation through effect of Fenton chemistry. Mass-to-charge ratio of identified oxalic acid from our published recent field study (carried out from same sampling location) was used for understanding the possible metallo-organic complex with Fe supports the substantial role of Fe in SOA formation in the deliquescent submicron particles facilitated by aqueous-phase chemistry.

Transition and heavy metals compared to oxidative parameter balance in patients with deep vein thrombosis: A case-control study.

Our study compared copper (Cu), zinc (Zn) and manganese (Mn), crucial for human normal physio-logy maintenance, and lead (Pb) levels as environmental pollutant, in subjects suffering of deep vein thrombosis of lower limbs (DVTs) vs. healthy subjects. Furthermore, we evaluated oxidative stress parameters, the thiobarbituric acid reactive substances (TBARS) as the sum of malondialdehyde or 1,1,3,3-tetraethoxypropane (MDA) and 4-hydroxynonenal 4-HNE) and cytosolic superoxide dismutase (Cu/Zn-SOD) concentrations in both groups. We recruited 24 DVT cases and 46 healthy subjects as controls. Questionnaire with socio­demographic, habits and lifestyle were collected. Hair concentrations of Zn, Mn, Cu and Pb were measured by ICP-MS, plasma concentrations of MDA and 4-HNE were measured by HPLC and SOD plasma concentrations were detected by ELISA test. A quantitative and qualitative variables comparison between cases and controls group was made by Mann-Whitney U test and Pearson's Chi-square test, respectively. We found low concentrations of Zn, Mn and Cu vs. high Pb concentrations in DVTs subjects. TBARS were found higher in the cases group, conversely, SOD concentrations were found lower in cases with respect to controls. Furthermore, we found the diet of pathological subjects significantly deficient in vegetables. These results are indicative of a lower enzymatic activity in patients, related to low transition metal levels in the DVTs and high levels of Pb, coupled with an unbalanced diet.

Health risk assessment of potentially harmful elements and dietary minerals from vegetables irrigated with untreated wastewater, Pakistan.

In the developing world, vegetables are commonly grown in suburban areas irrigated with untreated wastewater containing potentially harmful elements (PHEs). In Pakistan, there is no published work on the bioaccessibility aspect of PHEs and dietary minerals (DMs) in sewage-irrigated soil or the vegetables grown on such soils in Pakistan. Several industrial districts of Pakistan were selected for assessment of the risk associated with the ingestion of vegetables grown over sewage-irrigated soils. Both the total and bioaccessible fraction of PHEs (Cd, Co, Cr, Ni, and Pb) and DMs (Fe, Cu, Mn, Zn, Ca, Mg, and I) in soils and vegetable samples were measured. The concentrations of these PHEs and DMs in sewage-irrigated and control soils were below published upper threshold limits. However, compared to control soils, sewage irrigation over the years decreased soil pH (7.7 vs 8.1) and enhanced dissolved organic carbon (1.8 vs 0.8 %), which could enhance the phyto-availability of PHEs and DMs to crops. Of the PHEs and DMs, the highest transfer factor (soil to plant) was noted for Cd and Ca, respectively. Concentrations of PHEs in most of the sewage-irrigated vegetables were below the published upper threshold limits, except for Cd in the fruiting portion of eggplant and bell pepper (0.06-0.08 mg/kg Cd, dry weight) at three locations in Gujarat and Kasur districts. The bioaccessible fraction of PHEs can reduce the context of dietary intake measurements compared to total concentrations, but differences between both measurements were not significant for Cd. Since the soils of the sampled districts are not overly contaminated compared to control sites, vegetables grown over sewage-irrigated soils would provide an opportunity to harvest mineral-rich vegetables potentially providing consumers 62, 60, 12, 104, and 63 % higher dietary intake of Cu, Mn, Zn, Ca, and Mg, respectively. Based on Fe and vanadium correlations in vegetables, it is inferred that a significant proportion of total dietary Fe intake could be contributed by soil particles adhered to the consumable portion of vegetables. Faecal sterol ratios were used to identify and distinguish the source of faecal contamination in soils from Gujranwala, Gujarat, and Lahore districts, confirming the presence of human-derived sewage biomarkers at different stages of environmental alteration. A strong correlation of some metals with soil organic matter concentration was observed, but none with sewage biomarkers.

Comparative study of species sensitivity distributions based on non-parametric kernel density estimation for some transition metals.

Transition metals in the fourth period of the periodic table of the elements are widely widespread in aquatic environments. They could often occur at certain concentrations to cause adverse effects on aquatic life and human health. Generally, parametric models are mostly used to construct species sensitivity distributions (SSDs), which result in comparison for water quality criteria (WQC) of elements in the same period or group of the periodic table might be inaccurate and the results could be biased. To address this inadequacy, the non-parametric kernel density estimation (NPKDE) with its optimal bandwidths and testing methods were developed for establishing SSDs. The NPKDE was better fit, more robustness and better predicted than conventional normal and logistic parametric density estimations for constructing SSDs and deriving acute HC5 and WQC for transition metals in the fourth period of the periodic table. The decreasing sequence of HC5 values for the transition metals in the fourth period was Ti > Mn > V > Ni > Zn > Cu > Fe > Co > Cr(VI), which were not proportional to atomic number in the periodic table, and for different metals the relatively sensitive species were also different. The results indicated that except for physical and chemical properties there are other factors affecting toxicity mechanisms of transition metals. The proposed method enriched the methodological foundation for WQC. Meanwhile, it also provided a relatively innovative, accurate approach for the WQC derivation and risk assessment of the same group and period metals in aquatic environments to support protection of aquatic organisms.

Determination of lithium and transition metals in Li1 Ni1/3 Co1/3 Mn1/3 O2 (NCM) cathode material for lithium-ion batteries by capillary electrophoresis.

In this work, we present a novel electrophoretic method that was developed for the determination of lithium and transition metals in LiNi1/3 Co1/3 Mn1/3 O2 cathode material after microwave digestion. The cations in the digested LiNi1/3 Co1/3 Mn1/3 O2 material were separated by CE and the element content was determined by UV/Vis detection. To characterize the precision of the measurements, the RSDs and concentrations were calculated and compared to those obtained with ICP-optical emission spectrometry (ICP-OES). Furthermore, a certified reference material (BCR 176R-fly ash) was investigated for all techniques. For active material components, the LOD and LOQ were determined. The LODs and LOQs for the metals determined by CE were as follows: lithium (LOD/LOQ): 17.41/62.70 µg/L, cobalt (LOD/LOQ): 348.4/1283 µg/L, manganese (LOD/LOQ): 540.2/2095 µg/L, and nickel (LOD/LOQ): 838.0/2982 µg/L. Recovery rates for lithium were in the range of 95-103%. It could be proven that with the new technique, the results for the determination of the lithium content of active material were comparable with those obtained by ICP-OES and ion chromatography. Furthermore, the recovery rates of the transition metals were determined to be between 96 and 110% by CE and ICP-OES.

ROS-generating/ARE-activating capacity of metals in roadway particulate matter deposited in urban environment.

In this study we investigated the possible causal role for soluble metal species extracted from roadway traffic emissions in promoting particulate matter (PM)-induced reactive oxygen species (ROS) production and antioxidant response element (ARE) promoter activation. To this end, these responses have been evaluated in alveolar macrophage and epithelial lung cells that have been exposed to 'Unfiltered', 'Filtered' and 'Filtered+Chelexed' water extracts of PM samples collected from the roadway urban environments of Thessaloniki, Milan and London. Except for Thessaloniki, our results demonstrate that filtration resulted in a minor decrease in ROS activity of the fine PM fraction, suggesting that ROS activity is attributed mainly to water-soluble PM species. In contrast to ROS, ARE activity was mediated predominantly by the water-soluble component of PM present in both the fine and coarse extracts. Further removal of metals by Chelex treatment from filtered water extracts showed that soluble metal species are the major factors mediating ROS and ARE activities of the soluble fraction, especially in the London PM extracts. Finally, utilizing step-wise multiple-regression analysis, we show that 87% and 78% of the total variance observed in ROS and ARE assays, respectively, is accounted for by changes in soluble metal concentration. Using a statistical analysis we find that As, Zn and Fe best predict the ROS-generating/ARE-activating capacity of the near roadway particulate matter in the pulmonary cells studied. Collectively, our findings imply that soluble metals present in roadside PM are potential drivers of both pro- and anti-oxidative effects of PM in respiratory tract.

COST action TD1407: network on technology-critical elements (NOTICE)--from environmental processes to human health threats.

The current socio-economic, environmental and public health challenges that countries are facing clearly need common-defined strategies to inform and support our transition to a sustainable economy. Here, the technology-critical elements (which includes Ga, Ge, In, Te, Nb, Ta, Tl, the Platinum Group Elements and most of the rare-earth elements) are of great relevance in the development of emerging key technologies-including renewable energy, energy efficiency, electronics or the aerospace industry. In this context, the increasing use of technology-critical elements (TCEs) and associated environmental impacts (from mining to end-of-life waste products) is not restricted to a national level but covers most likely a global scale. Accordingly, the European COST Action TD1407: Network on Technology-Critical Elements (NOTICE)-from environmental processes to human health threats, has an overall objective for creating a network of scientists and practitioners interested in TCEs, from the evaluation of their environmental processes to understanding potential human health threats, with the aim of defining the current state of knowledge and gaps, proposing priority research lines/activities and acting as a platform for new collaborations and joint research projects. The Action is focused on three major scientific areas: (i) analytical chemistry, (ii) environmental biogeochemistry and (iii) human exposure and (eco)-toxicology.

Residential heating contribution to level of air pollutants (PAHs, major, trace, and rare earth elements): a moss bag case study.

In areas with moderate to continental climates, emissions from residential heating system lead to the winter air pollution peaks. The EU legislation requires only the monitoring of airborne concentrations of particulate matter, As, Cd, Hg, Ni, and B[a]P. Transition metals and rare earth elements (REEs) have also arisen questions about their detrimental health effects. In that sense, this study examined the level of extensive set of air pollutants: 16 polycyclic aromatic hydrocarbons (PAHs), and 41 major elements, trace elements, and REEs using Sphagnum girgensohnii moss bag technique. During the winter of 2013/2014, the moss bags were exposed across Belgrade (Serbia) to study the influence of residential heating system to the overall air quality. The study was set as an extension to our previous survey during the summer, i.e., non-heating season. Markedly higher concentrations of all PAHs, Sb, Cu, V, Ni, and Zn were observed in the exposed moss in comparison to the initial values. The patterns of the moss REE concentrations normalized to North American Shale Composite and Post-Archean Australian Shales were identical across the study area but enhanced by anthropogenic activities. The results clearly demonstrate the seasonal variations in the moss enrichment of the air pollutants. Moreover, the results point out a need for monitoring of air quality during the whole year, and also of various pollutants, not only those regulated by the EU Directive.

Recognition- and reactivity-based fluorescent probes for studying transition metal signaling in living systems.

Metals are essential for life, playing critical roles in all aspects of the central dogma of biology (e.g., the transcription and translation of nucleic acids and synthesis of proteins). Redox-inactive alkali, alkaline earth, and transition metals such as sodium, potassium, calcium, and zinc are widely recognized as dynamic signals, whereas redox-active transition metals such as copper and iron are traditionally thought of as sequestered by protein ligands, including as static enzyme cofactors, in part because of their potential to trigger oxidative stress and damage via Fenton chemistry. Metals in biology can be broadly categorized into two pools: static and labile. In the former, proteins and other macromolecules tightly bind metals; in the latter, metals are bound relatively weakly to cellular ligands, including proteins and low molecular weight ligands. Fluorescent probes can be useful tools for studying the roles of transition metals in their labile forms. Probes for imaging transition metal dynamics in living systems must meet several stringent criteria. In addition to exhibiting desirable photophysical properties and biocompatibility, they must be selective and show a fluorescence turn-on response to the metal of interest. To meet this challenge, we have pursued two general strategies for metal detection, termed "recognition" and "reactivity". Our design of transition metal probes makes use of a recognition-based approach for copper and nickel and a reactivity-based approach for cobalt and iron. This Account summarizes progress in our laboratory on both the development and application of fluorescent probes to identify and study the signaling roles of transition metals in biology. In conjunction with complementary methods for direct metal detection and genetic and/or pharmacological manipulations, fluorescent probes for transition metals have helped reveal a number of principles underlying transition metal dynamics. In this Account, we give three recent examples from our laboratory and collaborations in which applications of chemical probes reveal that labile copper contributes to various physiologies. The first example shows that copper is an endogenous regulator of neuronal activity, the second illustrates cellular prioritization of mitochondrial copper homeostasis, and the third identifies the "cuprosome" as a new copper storage compartment in Chlamydomonas reinhardtii green algae. Indeed, recognition- and reactivity-based fluorescent probes have helped to uncover new biological roles for labile transition metals, and the further development of fluorescent probes, including ones with varied Kd values and new reaction triggers and recognition receptors, will continue to reveal exciting and new biological roles for labile transition metals.

Potential importance of transition metals in the induction of DNA damage by sperm preparation media.

STUDY QUESTION: What are the mechanisms by which the preparation of spermatozoa on discontinuous density gradients leads to an increase in oxidative DNA damage? SUMMARY ANSWER: The colloidal silicon solutions that are commonly used to prepare human spermatozoa for assisted reproduction technology (ART) purposes contain metals in concentrations that promote free radical-mediated DNA damage. WHAT IS KNOWN ALREADY: Sporadic reports have already appeared indicating that the use of colloidal silicon-based discontinuous density gradients for sperm preparation is occasionally associated with the induction of oxidative DNA damage. The cause of this damage is however unknown. STUDY DESIGN, SIZE, DURATION: This study comprised a series of experiments designed to: (i) confirm the induction of oxidative DNA damage in spermatozoa prepared on commercially available colloidal silicon gradients, (ii) compare the levels of damage observed with alterative sperm preparation techniques including an electrophoretic approach and (iii) determine the cause of the oxidative DNA damage and develop strategies for its prevention. The semen samples employed for this analysis involved a cohort of >50 unselected donors and at least three independent samples were used for each component of the analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: The setting was a University biomedical science laboratory. The major techniques employed were: (i) flow cytometry to study reactive oxygen species generation, lipid peroxidation and DNA damage, (ii) computer-aided sperm analysis to measure sperm movement and (iii) inductively coupled mass spectrometry to determine the elemental composition of sperm preparation media. MAIN RESULTS AND THE ROLE OF CHANCE: Oxidative DNA damage is induced in spermatozoa prepared on PureSperm(®) discontinuous colloidal silicon gradients (P < 0.001 versus repeated centrifugation) because this medium contains metals, particularly Fe, Al and Cu, which are known to promote free radical generation in the immediate vicinity of DNA. This damage can be significantly accentuated by reducing agents, such as ascorbate (P < 0.001) and inhibited by selective chelation (P < 0.001). This problem is not confined to PureSperm(®); analysis of additional commercial sperm preparation media revealed that metal contamination is a relatively constant feature of such products. LIMITATIONS, REASONS FOR CAUTION: While the presence of metals, particularly transition metals, may exacerbate the levels of oxidative DNA damage seen in human spermatozoa, the significance of such damage has not yet been tested in suitably powered clinical trials. WIDER IMPLICATIONS OF THE FINDINGS: The results explain why the preparation of spermatozoa on discontinuous colloidal silicon gradients can result in oxidative DNA damage. The results are of immediate relevance to the development of safe, effective protocols for the preparation of spermatozoa for ART purposes. STUDY FUNDING/COMPETING INTERESTS: The study was funded by the Australian Health and Medical Research Council. One of the authors (R.J.A.) has had a consultantship with a biotechnology company, NuSep, interested in the development of electrophoretic methods of sperm preparation. He has no current financial interest in this area. None of the other authors have a conflict of interest to declare.

ON-OFF mechanism of a fluorescent sensor for the detection of Zn(II), Cd(II), and Cu(II)transition metal ions.

An ab initio multiconfigurational (CASPT2//CASSCF) approach has been employed to map radiative and nonradiative relaxation pathways for a cyclam-methylbenzimidazole fluorescent sensor and its metal ion (Zn(2+), Cd(2+), and Cu(2+)) complexes to provide an universal understanding of ON-OFF fluorescent mechanisms for the selective identification of these metal ions. The photoinduced electron transfer (PET) between the receptor and the signaling unit is quantitatively attributed for the first time to a newly generated transition of S0→SCT((1)nπ*), which is a typical (1)nπ* excitation but exhibits a significant charge transfer character and zwitterionic radical configuration. The present study contributes the two theoretical models of the competitive coexistence of radiative/nonradiative decay channel in (1)ππ*/SCT((1)nπ*) states for the detection of metal ions with d(10) configuration (i.e., Zn(2+), Cd(2+), etc.) and a downhill ladder relaxation pathway through multi nona-diabatic relays for the probing of d(9) cations (Cu(2+), etc.). These computational results will establish a benchmark for ON-OFF mechanisms of a fluorescent sensor that coordinates various transition metal ions with different electron configuration and radius.