Determination of ruthenium on DNA by XRF

An X-ray fluorescence (XRF) technique is used to quantitate the binding of [H₂O(NH₃)₅Ru^II]²⁺ to DNA. This method is shown to be more sensitive, precise, and convenient than conventional optical absorption (OA) spectroscopy, differential pulse voltammetry (DPV), or atomic absorption (AA) techniques. X-ray fluorescence is insensitive to the oxidation state or coordination environment of Ru and so can be used to determine total Ru. The minimum detectable amount of Ru is 10 ng in 1 h of counting time, using a 100-mCi¹²⁵I source. The specific advantages of the XRF method over the conventional methods are outlined.     

Intracellular sequestration of manganese and phosphorus in a metal-resistant fungus <Emphasis Type="Italic">Cladosporium cladosporioides</Emphasis> from deep-sea sediment

A heavy metal resistant fungus was isolated from the sediment of Pacific Ocean, and identified to be Cladosporium cladosporioides. It grew normally in a medium containing 60 mM Mn²⁺ and could endure 1,200 mM as the highest concentration tested. Quantification analysis confirmed a high accumulation of Mn which was 58 mg/g in dried biomass. Under transmission electron microscope, many intracellular crystals were observed in the cytoplasm of the hypha cells grown in a Mn-rich medium, and varied from a few nanometers to 200 nm in length. Energy dispersive X-ray (EDX) analysis showed that the crystals were composed of manganese and phosphorus in atomic ratio of 1.6:1 (Mn/P). Further, factors which might influence the resistance of this fungus were investigated. As a result, its high resistance to Mn²⁺ was found dependent on the presence of Mg²⁺, and could be further enhanced by phosphate. However, the effect of phosphate was not observed without the presence of Mg²⁺. In addition, the resistance was also influenced by pH of the medium, which was lost above pH 8. This is the first report on a fungus which showed a hyper resistance to manganese by forming a large quantity of intracellular Mn/P crystals.     

Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells

It remains unexplored in what way osteogenic stimulation with dexamethasone, ascorbic acid and β-glycerol phosphate (DAG) influences the process of mineralization, the composition and structure of the assembled mineral. Therefore, we analyzed and characterized biomineralization in DAG-stimulated and unstimulated 3D human unrestricted somatic stem cell (USSC) cultures. Microspheres were analyzed by histological staining, scanning electron microscopy (SEM), semi-quantitative energy-dispersive X-ray spectroscopy (EDX), quantitative wavelength-dispersive X-ray spectroscopy (WDX), transmission electron microscopy (TEM), selected area electron diffraction (SAED) and Raman spectroscopy.Mineral material was detected by SEM and histological staining in both groups, and showed structural differences. DAG influenced the differentiation of USSCs and the formation, structure and composition of the assembled mineral. SEM showed that cells of the + DAG spheres exhibited morphological signs of osteoblast-like cells. EDX and WDX confirmed a Ca-P mineral in both groups. Overall, the mineral material found showed structural similarities to the mineral substance of bony material.     

Metal Content in Air Samples Collected in an Urban Zone in Tampico,México: A First Survey

With the aim to know possible risks to the population, Cd, Co, Cr, Cu, Mn, Ni, Pb, and Tl were determined for the first time in airborne samples of particulate matter in an urban zone in Tampico, México, during the winter of 2003. The 24-hour PM₁₀ samples were collected every 6 days on quartz-filters by using a high volume sampler and analyzed by Inductively Coupled Plasma Optical Emission Spectrometer. Standard reference material was used to verify metal recovery. The maximum PM₁₀ and lead concentrations were 12.05 and 0.040 μ g/m³, respectively, not exceeding Méxican standard values. The greatest metal concentration was that of manganese with 0.90 μ g/m³, followed by Cu and Ni with 0.17 and 0.012 μ g/m³, respectively. Agglomerates, well-defined particles, and heavy metals (e.g., Mn and Cu) were found in PM₁₀ using Scanning Electron Microscopy and Energy Dispersive Spectroscopy. Meteorological conditions associated with the sampling period showed that Pb and Ni are being continuously emitted, and that Mn, Cu, and Co could come from one industry located to the WSW of the region. All of these concentrations do not constitute a potential risk to human health, although it is necessary to continue studying the high concentrations of Mn and Cu in longer sampling periods.     

How much manganese is safe for infants? A review of the scientific basis of intake guidelines and regulations relevant to the manganese content of infant formulas

BackgroundRecent research has uncovered the potential for excess manganese (Mn) intakes causing significant neurotoxic effects for early brain development.MethodsWe identified the Mn tolerable intakes (TI) published by the U.S. Institute of Medicine (IOM), World Health Organization (WHO), Agence nationale de sécurité sanitaire (ANSES), and U.S. Environmental Protection Agency (US EPA) and examined the primary studies on which regulatory TIs are based. We converted the TIs to μg of Mn/kg/day using standard assumptions specific to each agency. We estimated μg of Mn/kg/day intakes due to formulas. Using our estimates for formula intakes, weights, and kcal content, we converted regulatory maxima and minima from μg of Mn/100 kcals to estimates of μg of Mn/kg/day.ResultsExcept for the proposed ANSES TI for drinking water, none of the primary studies on which Mn intake guidelines and regulations are based measured health outcomes. Some infant formulas may exceed the regulatory TIs, especially if prepared with water containing considerable concentrations of Mn (e.g. 250 μg/L), even while meeting national and international regulatory standards or guidelines.ConclusionsInfant formula regulations must be revised to reduce the potential for excess manganese intakes and the practice of manganese supplementation of infant formulas should be ceased.     

Distribution of metals in the termite Tumulitermes tumuli (Froggatt): two types of Malpighian tubule concretion host Zn and Ca mutually exclusively

The aim of this study was to determine specific distribution of metals in the termite Tumulitermes tumuli (Froggatt) and identify specific organs within the termite that host elevated metals and therefore play an important role in the regulation and transfer of these back into the environment. Like other insects, termites bio-accumulate essential metals to reinforce cuticular structures and utilize storage detoxification for other metals including Ca, P, Mg and K. Previously, Mn and Zn have been found concentrated in mandible tips and are associated with increased hardness whereas Ca, P, Mg and K are accumulated in Malpighian tubules. Using high resolution Particle Induced X-Ray Emission (PIXE) mapping of whole termites and Scanning Electron Microscope (SEM) Energy Dispersive X-ray (EDX) spot analysis, localised accumulations of metals in the termite T. tumuli were identified. Tumulitermes tumuli was found to have proportionally high Mn concentrations in mandible tips. Malpighian tubules had significant enrichment of Zn (1.6%), Mg (4.9%), P (6.8%), Ca (2.7%) and K (2.4%). Synchrotron scanning X-ray Fluorescence Microprobe (XFM) mapping demonstrated two different concretion types defined by the mutually exclusive presence of Ca and Zn. In-situ SEM EDX realisation of these concretions is problematic due to the excitation volume caused by operating conditions required to detect minor amounts of Zn in the presence of significant amounts of Na. For this reason, previous researchers have not demonstrated this surprising finding.     

Estimating daily intakes of manganese due to breast milk,infant formulas,or young child nutritional beverages in the United States and France: Comparison to sufficiency and toxicity thresholds

BackgroundAlthough manganese (Mn) is an essential nutrient, recent research has revealed that excess Mn in early childhood may have adverse effects on neurodevelopment.MethodsWe estimated daily total Mn intake due to breast milk at average body weights by reviewing reported concentrations of breast milk Mn and measurements of body weight and breast milk intake at 3 weeks, 4.25 months, 7 months, and 18 months. We compared these figures to the Mn content measured in 44 infant, follow-up, and toddler formulas purchased in the United States and France. We calculated Mn content of formula products made with ultra-trace elemental analysis grade water (0 μg Mn/L) and with water containing 250 μg Mn/L, a concentration which is relatively high but less than the World Health Organization Health-based value of 400 μg Mn/L or the United States Environmental Protection Agency Health Advisory of 350 μg Mn/L.ResultsEstimated mean daily Mn intake from breast milk ranged from 1.2 μg Mn/kg/day (3 weeks) to 0.16 μg Mn/kg/day (18 months), with the highest intakes at the youngest age stage we considered, 3 weeks. Estimated daily Mn intake from formula products reconstituted with 0 μg Mn/L water ranged from 130 μg Mn/kg/day (3 weeks) to 4.8 μg Mn/kg/day (18 months) with the highest intakes at 3 weeks. Formula products provided 28–520 times greater than the mean daily intake of Mn from breast milk for the 4 age stages that we considered. Estimated daily Mn intake from formula products reconstituted with water containing 250 μg Mn/L ranged from 12 μg Mn/kg/day to 170 μg Mn/kg/day, which exceeds the United States Environmental Protection Agency Reference Dose of 140 μg Mn/kg/day for adults.ConclusionsMn deficiency is highly unlikely with exclusive breast milk or infant formula feeding, but established tolerable daily intake levels for Mn may be surpassed by some of these products when following labeled instructions.     

Ethanol inhibits leptin-induced STAT3 activation in Huh7 cells

The manganese (Mn) complex of photosystem II catalyzes water oxidation. For the first time, its advancement through the reaction cycle was monitored by time-resolved X-ray absorption measurements at the Mn K-edge at room temperature. The complex was stepped through its four oxidation states by nano-second-laser flashes applied to samples exposed to the X-ray beam. Time courses of the X-ray fluorescence intensity were recorded during a flash sequence. Extended X-ray absorption fine-structure spectra were recorded with the S(1), S(2), and S(3) oxidation states highly populated. The room temperature data is compatible with the formation of a third di-mu-oxo bridge between two Mn atoms upon the S(2)-->S(3) transition.     

Detection of airborne lactococcal bacteriophages in cheese manufacturing plants

The dairy industry adds starter bacterial cultures to heat-treated milk to control the fermentation process during the manufacture of many cheeses. These highly concentrated bacterial populations are susceptible to virulent phages that are ubiquitous in cheese factories. In this study, the dissemination of these phages by the airborne route and their presence on working surfaces were investigated in a cheese factory. Several surfaces were swabbed, and five air samplers (polytetrafluoroethylene filter, polycarbonate filter, BioSampler, Coriolis cyclone sampler, and NIOSH two-stage cyclone bioaerosol personal sampler) were tested. Samples were then analyzed for the presence of two Lactococcus lactis phage groups (936 and c2), and quantification was done by quantitative PCR (qPCR). Both lactococcal phage groups were found on most swabbed surfaces, while airborne phages were detected at concentrations of at least 10(3) genomes/m(3) of air. The NIOSH sampler had the highest rate of air samples with detectable levels of lactococcal phages. This study demonstrates that virulent phages can circulate through the air and that they are ubiquitous in cheese manufacturing facilities.     

Mn exerts stronger structural effects than the Mn-citrate complex on the human erythrocyte membrane and molecular models

While traces of manganese (Mn) take part in important and essential functions in biology, elevated exposures have been shown to cause significant toxicity. Chronic exposure to the metal leads to manganese neurotoxicity (or manganism), a brain disorder that resembles Parkinsonism. Toxic effect mechanisms of Mn is not understood, toxic concentrations of manganese are not well defined and blood manganese concentration at which neurotoxicity occurs has not been identified. There are reports indicating that the most abundant Mn-species in Mn carriers within blood is the Mn-citrate complex. Despite the well-documented information about the toxic effects of Mn, there are scarce reports concerning the effects of manganese compounds on both structure and functions of cell membranes, particularly those of human erythrocytes. With the aim to better understand the molecular mechanisms of the interaction of Mn with cell membranes, MnCl₂, and the Mn-citrate complex were incubated with intact erythrocytes, isolated unsealead human erythrocyte membranes (IUM), and molecular models of the erythrocyte membrane. These consisted in bilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), phospholipid classes present in the outer and inner monolayers of the erythrocyte membrane, respectively. The capacity of the Mn compounds to perturb the bilayer structures of DMPC and DMPE was evaluated by X-ray diffraction, IUM were studied by fluorescence spectroscopy, and intact human erythrocytes were observed by scanning electron microscopy (SEM). In all these systems it was found that Mn²⁺ exerted considerable higher structural perturbations than the Mn-citrate complex.     

More manganese accumulates in maple sun leaves than in shade leaves

Nuclear magnetic resonance and neutron activation analysis were used to measure manganese concentrations in leaves of Acer platanoides. Mn was found to accumulate in both the vacuoles and the chloroplasts, with more Mn (per unit area) in sun leaves than in shade leaves. No Mn was lost at senescence. Different seasonal patterns of Mn accumulation were found in sun and shade leaves. The quantity of chloroplast reserve Mn (bound to the outer surface of thylakoid membranes) increased rapidly in sun leaves from bud-break through midsummer, and then remained approximately constant through senescence. In shade leaves, however, the quantity of reserve Mn increased slowly, and at approximately a constant rate throughout the growing season.     

Bone calcium/phosphorus ratio determination using dual energy X-ray method

Non-invasive dual energy methods have been used extensively on osteoporosis diagnosis estimating parameters, such as, Bone Mineral Density (BMD) and Bone Mineral Content (BMC). In this study, an X-ray dual energy method (XRDE) was developed for the estimation of the bone Calcium-to-Phosphorous (Ca/P) mass ratio, as a bone quality index. The optimized irradiation parameters were assessed by performing analytical model simulations. X-ray tube output, filter material and thickness were used as input parameters. A single exposure technique, combined with K-edge filtering, was applied. The optimal X-ray spectra were selected according to the resulted precision and accuracy values. Experimental evaluation was performed on an XRDE system incorporating a Cadmium Telluride (CdTe) photon counting detector and three bone phantoms with different nominal mass Ca/P ratios. Additionally, the phantoms’ mass Ca/P ratios were validated with energy-dispersive X-ray spectroscopy (EDX). Simulation results showed that the optimum filter atomic number (Z) ranges between 57 and 70. The optimum spectrum was obtained at 100 kVp, filtered with Cerium (Ce), with a surface density of 0.88 g/cm². All Ca/P ratio measurements were found to be accurate to within 1.6% of the nominal values, while the precision ranged between 0.91 and 1.37%. The accuracy and precision values of the proposed non-invasive method contributes to the assessment of the bone quality state through the mass Ca/P ratio determination.     

The Protective Effects of Riluzole on Manganese-Induced Disruption of Glutamate Transporters and Glutamine Synthetase in the Cultured Astrocytes

Chronic exposure to excessive manganese (Mn) can lead to manganism, a type of neurotoxicity accomplished with extracellular glutamate (Glu) accumulation. To investigate this accumulation, this study focused on the role of astrocyte glutamate transporters (GluTs) and glutamine synthetase (GS), which have roles in Glu transport and metabolism, respectively. And the possible protective effects of riluzole (a glutamatergic modulator) were studied in relation to Mn exposure. At first, the astrocytes were exposed to 0, 125, 250, and 500 μM MnCl(2) for 24 h, and 100 μM riluzole was pretreated to astrocytes for 6 h before 500 μM MnCl(2) exposure. Then, [(3)H]-glutamate uptake was measured by liquid scintillation counting; Na(+)-K(+) ATPase and GS activities were determined by a colorimetric method; glutamate/aspartate transporter (GLAST), glutamate transporter-1 (GLT-1), and GS mRNA expression were determined by RT-PCR and protein levels were measured by western blotting. The results showed that Mn inhibited Glu uptake, Na(+)-K(+) ATPase and GS activities, GLAST, GLT-1, and GS mRNA, and protein in a concentration-dependent manner. And they were significantly higher for astrocytes pretreated with 100 μM riluzole than the group exposed to 500 μM MnCl(2). The results suggested that Mn disrupted Glu transport and metabolism by inhibiting GluTs and GS. Riluzole activated protective effects on enhancing GluTs and GS to reverse Glu accumulation. In conclusion, Mn exposure results in the disruption of GLAST, GLT-1, and GS expression and function. Furthermore, riluzole attenuates this Mn toxicity.     

Vitamin C in premature and full-term human neonates

Summary

Plasma concentrations of vitamin C (ascorbic acid, AA) are known to be higher in full-term human neonates than their mothers. Immaturity of placental AA transport could result in low plasma AA concentrations in pre-term infants. We found that plasma AA concentrations in umbilical cord blood of 25 full-term neonates (38–42 weeks gestation) and 33 pre-term neonates (24–36 weeks gestation) were always significantly higher than in the corresponding maternal blood (P < 0.0001). However, plasma AA levels were significantly higher in pre-term than in full-term infants (146 ± 93 vs 102 ± 27 μM, respectively; P = 0.03). Furthermore, a rapid and sharp decrease in plasma AA concentrations from 229 ± 166 μM to 45 ± 18 μM (P < 0.0001) over the first 3 days of life was observed in eight very low birth weight infants (460–1090 g, 24–28 weeks gestation). These findings raise important questions about the in utero functions of AA in the developing fetus and the adequacy of postnatal vitamin C supplementation of the premature infant.     

Effect of manganese toxicity on growth and N2 fixation in groundnut, Arachis hypogaea

The development of manganese (Mn) toxicity symptoms and its effects on the growth, nodulation, and nitrogen fixation of groundnut genotypes were examined using a quartz-sand/solution culture system. The 11 genotypes tested all accumulated considerable concentrations of manganese (1.04–3.07 mg g^-1dry matter) when supplied with 15 μg Mn ml^-1of nutrient solution daily. Toxicity symptoms differed between genotypes: some showed no visual effects, some produced marginal leaf spots, and others developed marginal leaf spots coupled with an inward rolling of the margins of the younger leaves. The growth of one genotype (ICG 5394) grown with inorganic nitrogen as its source of N was more severely affected by Mn toxicity than when dependent on symbiotic fixation for its nitrogen.     

Isolierung und Zählung von Bodenactinomyceten auf Erdplatten mit Membranfiltern

Summary A technique is described to separate actinomycetes from other micro-organisms with membrane filters on soil plates. A measured quantity of soil dilution was passed through a membrane filter and the filter with the adhering organisms was placed upside down on a medium provided with soil. After an incubation period of two weeks all actinomycetes had penetrated the filter developing colonies on the sterile surface, whereas the other organisms were retained by the filter. By this procedure the counting and isolation of actinomycetes has been facilitated. The selection of bacteriostatic active forms is practicable by stamping the crowded filter on a medium that contains test bacteria.

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Nutrient concentrations of 17- year-old Pinus taeda annual tree-rings analyzed by X-ray fluorescence microanalysis

Tree-rings are sensitive indicators of soil chemical changes. X-ray fluorescence microanalysis (μ-XRF) can reveal the elemental distribution pattern along these rings. However, reports on quantitative μ-XRF methods targeted to wood analysis are scarce. This study aimed to analyze iron (Fe), manganese (Mn), calcium (Ca), potassium (K), sulfur (S) and phosphorus (P) in annual tree-rings of wood cores cut from 24 trees of 17 year-old Pinus taeda planted in soil amended with six doses of composted pulp-mill sludge (CPMS). The nutrient concentrations were accessed using calibration curves built with spiked P. taeda wood pellets. Calcium and Mn content decreased from the pith to bark direction; K and S decreased from the pith up to 3rd tree-ring and, then, increased to the bark. Iron and P slight decreased from the pith up to the 13–14th tree-ring. Calcium, K and S presented strong and positive correlation with the rainier and hotter season (r > 0.4, p < 0.05). The CPMS increased the Ca, K, Fe and S and decreased Mn and P concentration in P. taeda wood in the 2nd–5th years. Furthermore, the P. taeda annual tree-ring molar ratios of Ca/Mn and K/Ca were good indicators of soil-pH and wood cambium activity. The μ-XRF methodology, as non-destructive method of nutrient concentration analysis in tree-rings, revealed potential uses in monitoring soil fertilizer treatments.     

Structural Influences of Sodium and Calcium Ions on the Biogenic Manganese Oxides Produced by the Marine Bacillus Sp., Strain SG-1

Natural manganese oxide nanoparticles and grain coatings profoundly impact contaminant cycling in the environment through their ability to degrade organic compounds and sequester metal ions. Previous studies of biogenic manganese oxides have shown that the interlayer cation may have an important effect on the resulting oxide structure. The effect of Na and Ca ions was investigated to determine their fundamental roles in the stabilization of the phyllomanganate biooxide structure, its unit cell symmetry, and order/disorder relations. Biogenic oxides were created by incubating Mn(II) with spores of the marine Bacillus sp., strain SG-1 and the resulting oxide structures examined using X-ray absorption spectroscopy and X-ray diffraction to determine the short-range and long-range atomic structure. Phyllomanganates were observed exclusively, with differing degrees of layer stacking disorder, degree of crystallinity, and layer symmetry, depending on the cation present. In general, Ca was found to promote biooxide long-range order. We conclude that the presence of Ca in these oxides will confer greater stability to these bacteriogenic manganese bioxodes.     

A comparative study of hepatic trace element levels in wild moose, roe deer, and reindeer from Norway

Liver samples from 422 wild moose (Alces alces), 280 roe deer (Capreolus capreolus), and 73 reindeer (Rangifer tarandus) collected by hunters in various localities in Norway, 2002-2003, were analyzed for the essential trace elements cobalt, copper (Cu), manganese (Mn), molybdenum, selenium (Se), and zinc. Significant differences in hepatic concentrations among species were found for all elements except for Mn, and considerable individual and geographic variations were seen. Roe deer had statistically significant lower Se levels (median: 0.51 μg Se/g dry weight) than did moose (0.77 μg Se/g) and reindeer (0.85 μg Se/g). Moose from two coastal municipalities with high precipitation had considerably higher Se concentrations than those from the other localities studied. Seventy-nine roe deer (28%) and 36 moose (9%) had Se concentrations below that regarded as deficient in domestic ruminants. The Se status in roe deer was lower than previously reported in Europe. Moose had a significantly higher Cu (222 μg Cu/g dw) than did roe deer (112 μg Cu/g) and reindeer (105 μg Cu/g). The Cu status of moose and roe deer in Norway are among the highest reported in Europe. However, a suboptimal Se and Cu status was found in moose from Tvedestrand, a population which has suffered from a reduced condition and productivity. The variability in trace element status among hunted cervids, with no apparent signs of deficiency or toxicity, probably reflects adaptations in these wild ruminant species to cope with this. However, subtle clinical signs and lesions are difficult to detect and further research is needed.     

Investigation of Cr(VI) adsorption onto chemically treated Helianthus annuus: Optimization using Response Surface Methodology

In the present study, chemically treated Helianthus annuus flowers (SHC) were used to optimize the removal efficiency for Cr(VI) by applying Response Surface Methodological approach. The surface structure of SHC was analyzed by Scanning Electron Microscopy (SEM) coupled with Energy Dispersive X-ray Analysis (EDX). Batch mode experiments were also carried out to assess the adsorption equilibrium in aqueous solution. The adsorption capacity (q_e) was found to be 7.2 mg/g. The effect of three parameters, that is pH of the solution (2.0-7.0), initial concentration (10-70 mg/L) and adsorbent dose (0.05-0.5 g/100 mL) was studied for the removal of Cr(VI) by SHC. Box-Behnken model was used as an experimental design. The optimum pH, adsorbent dose and initial Cr(VI) concentration were found to be 2.0, 5.0 g/L and 40 mg/L, respectively. Under these conditions, removal efficiency of Cr(VI) was found to be 90.8%.