The Effect of Age on 12 Chemical Element Contents in the Intact Prostate of Adult Men Investigated by Inductively Coupled Plasma Atomic Emission Spectrometry

The effect of age on 12 chemical element contents in intact prostate of 64 apparently healthy, 13-60-year-old men (mean age 36.5 years) was investigated by inductively coupled plasma atomic emission spectrometry. Mean values (M ± SΕΜ) for mass fraction (milligrams/kilogram, on dry weight basis) of Ba, Ca, Cu, Fe, K, Mg, Na, P, S, Sr, and Zn were: Ba 1.18 ± 0.12, Ca 2,178 ± 160, Cu 10.7 ± 0.9, Fe 122 ± 5, K 12,530 ± 360, Mg 1,100 ± 70, Na 10,470 ± 320, P 7,580 ± 300, S 8,720 ± 180, Sr 1.85 ± 0.28, and Zn 782 ± 97, respectively. The upper limit of mean content of V was ≤0.22 mg/kg. A tendency of age-related increase in Ca, Fe, Na, and Zn mass fraction as well an increase in Zn/Ba, Zn/Ca, Zn/Cu, Zn/Fe, Zn/K, Zn/Mg, Zn/Na, Zn/P, Zn/S, and Zn/Sr ratios in prostate was observed. A significant positive correlation was seen between the prostatic zinc and Ca, Cu, Fe, Mg, Na, and P contents.     

Distribution of N,P, K,Ca, Mg,S, Zn,Fe, Mn and Cu in groundnut

Summary Concentration of N, P, K, Ca, Mg and S in summer groundnut crop was higher than in kharif while Zn, Fe, Mn and Cu contents were higher in summer crop. Kernel's N, P and Zn; Leaflet's Ca and Mn; Stem's K and Fe; Root's S and Cu and Petiole's Mg contents were highest. Shell's N, P, K, Mg, S, Zn and Cu; Kernel's Ca, Fe and Mn contents were the least. N, P, K, S, Zn and Cu concentrations decreased linearly as the crop grew. Ca, Mg, Fe and Mn concentrations did not display any distinct pattern. Ca concentration was positively correlated with pod yield in both the seasons.     

The elemental composition of the chloragosomes of two earthworm species (Lumbricus terrestris and Allolobophora longa) determined by electron probe X-ray microanalysis of freeze-dired cryosections

Summary The elemental composition of the chloragosomes of the two earthworm species, one with complex highly active Ca-secreting calciferous glands (L. terrestris) and the other with non-secretory glands (A. longa), was determined by the electron probe X-ray microanalysis of unfixed freeze-dried cryosections. The predominant constituents of the chloragosomes of both species were P, Ca, Zn and S, with lesser quantities of K, Cl and Fe. The most striking species differences in chloragosomal chemistry were the higher concentrations (expressed as relative mass fractions) of P(×2.1), Ca(×1.5), and Zn(×2.3) in L. terrestris, and the much higher S(×19.6) in A. longa. These differences were discussed in relation to the general ecophysiology of the two species, and more specifically in relation to heavy metal uptake and binding.     

Remarkable effect of mobile phase buffer on the SEC-ICP-AES derived Cu, Fe and Zn-metalloproteome pattern of rabbit blood plasma

The development of an analytical method to quantify the major Cu, Fe and Zn-containing metalloproteins in mammalian plasma has been recently reported. This method is based on the separation of plasma proteins by size exclusion chromatography (SEC) followed by the on-line detection of the metalloproteins by an inductively coupled plasma atomic emission spectrometer (ICP-AES). To assess whether the mobile phase buffer can affect the SEC-ICP-AES-derived metalloproteome pattern, thawed rabbit plasma was analyzed using phosphate buffered saline (PBS)-buffer (0.15 M, pH 7.4), Tris-buffer (0.1 and 0.05 M, pH 7.4), Hepes-buffer (0.1 M, pH 7.4) or Mops-buffer (0.1 M, pH 7.4). In contrast to the Cu-specific chromatograms, the Fe and Zn-specific chromatograms that were obtained with Tris, Hepes and Mops-buffer were considerably different from those attained with PBS-buffer. The Tris, Hepes and Mops-buffer mediated redistribution of ~25% plasma Zn(2+) from <100 kDa to >100-600 kDa plasma proteins and to a smaller extent to a <10 kDa (Tris)(2)Zn(2+)-complex can be rationalized in terms of the abstraction of Zn(2+) from the weak binding site on albumin. In contrast, only Hepes and Mops-buffer redistributed ~20% of plasma Fe(3+) from the <100 kDa to the >600 kDa elution range. Based on these results and considering that the utilization of PBS-buffer has previously resulted in the detection of a number of Cu, Fe and Zn-containing metalloentities in rabbit plasma that was most consistent with literature data, this mobile phase buffer is recommended for metallomic studies regarding mammalian blood plasma.     

Medicago sativa - Sinorhizobium meliloti Symbiosis Promotes the Bioaccumulation of Zinc in Nodulated Roots

In this study we investigated effects of Zn supply on germination, growth, inorganic solutes (Zn, Ca, Fe, and Mg) partitioning and nodulation of Medicago sativa This plant was cultivated with and without Zn (2 mM). Treatments were plants without (control) and with Zn tolerant strain (S532), Zn intolerant strain (S112) and 2 mM urea nitrogen fertilisation. Results showed that M. sativa germinates at rates of 50% at 2 mM Zn. For plants given nitrogen fertilisation, Zn increased plant biomass production. When grown with symbionts, Zn supply had no effect on nodulation. Moreover, plants with S112 showed a decrease of shoot and roots biomasses. However, in symbiosis with S532, an increase of roots biomass was observed. Plants in symbiosis with S. meliloti accumulated more Zn in their roots than nitrogen fertilised plants. Zn supply results in an increase of Ca concentration in roots of fertilised nitrogen plants. However, under Zn supply, Fe concentration decreased in roots and increased in nodules of plants with S112. Zn supply showed contrasting effects on Mg concentrations for plants with nitrogen fertilisation (increase) and plants with S112 (decrease). The capacity of M. sativa to accumulate Zn in their nodulated roots encouraged its use in phytostabilisation processes.     

镉胁迫对秋华柳根系活力及其Ca、Mg、Mn、Zn、Fe积累的影响

以秋华柳为试验材料,采用水培试验方式,设置CK(0 mg·L^-1 Cd²⁺)、T₁(2 mg·L^-1 Cd²⁺)、T₂(10 mg·L^-1Cd²⁺)、T₃(20 mg·L^-1Cd²⁺)、T₄(50 mg·L^-1Cd²⁺)5种镉处理浓度,通过对秋华柳根系活力,叶、韧皮部、木质部和根部的镉含量,以及Ca、Mg、Mn、Zn、Fe 5种常规金属元素含量的测定,研究了不同浓度镉胁迫下秋华柳根系活力及Ca、Mg、Mn、Zn、Fe 等金属元素含量的变化.结果表明:1)秋华柳根系活力随着镉处理浓度的增加而逐渐下降,当镉浓度≥10 mg·L^-1时,根系活力与对照相比显著下降.2)随着镉处理浓度的增加,秋华柳叶中Fe的积累受到显著抑制;韧皮部Mg、Mn、Fe的积累受到显著抑制;木质部Ca、Mg、Mn、Zn、Fe 等5种常规金属元素的积累无显著性差异;根部5种常规金属元素的吸收和积累受到显著抑制,表现出镉对其他金属元素积累的拮抗作用.其中,50 mg·L^-1的镉胁迫下,根部Zn的积累量降幅最大,受Cd的抑制最明显.3)各处理组Fe的转移系数与对照相比均无显著性差异;Ca、Mg、Mn、Zn的转移系数均高于对照,且在一定的镉处理水平上差异显著.4)镉胁迫下,秋华柳根部镉的积累量与Ca的积累量呈显著负相关,与其他4种常规金属元素的积累量呈极显著负相关,说明根部常规金属元素的变化可作为秋华柳受镉毒害程度的指示之一.     

Trace elements in the teeth of Steller sea lions (<Emphasis Type="Italic">Eumetopias jubatus</Emphasis>) from the North Pacific

The occurrence of trace elements in the teeth of Steller sea lions (Eumetopias jubatus) collected from the North Pacific from 1968 to 1999 is described. Using particle-induced X-ray emission analysis, we detected eight trace elements: Ca, Mn, Fe, Cu, Zn, Br, Sr and Pb. We used a general linear model to evaluate long-term trends of trace element accumulation. Zn (Zn/Ca) and Fe (Fe/Ca) levels both varied during the last three decades, and showed significant differences between animals of different age (Fe/Ca), season (Zn/Ca and Fe/Ca), and sex (Zn/Ca); both Zn/Ca and Fe/Ca ratios were higher in young males than older males. Pb was detected in 39% of the samples from Hokkaido (Japan) and 22% of those from Alaska. The high levels of Pb recorded in 1975 were thought to be related to the use of leaded gasoline during the 1970s. Our results indicate that the levels of trace elements in the teeth of Steller sea lions can be used as indicators of temporal and spatial variations of trace element pollution.     

Simultaneous Cu-, Fe-, and Zn-specific detection of metalloproteins contained in rabbit plasma by size-exclusion chromatography–inductively coupled plasma atomic emission spectroscopy

Analytical methods which are capable of determining the plasma or serum metalloproteome have inherent diagnostic value for human diseases associated with increased or decreased concentrations of specific plasma metalloproteins. We have therefore systematically developed a method to rapidly determine the major Cu-, Fe-, and Zn-containing metalloproteins in rabbit plasma (0.5 mL) based on size-exclusion chromatography (SEC; stationary phase Superdex 200, mobile phase phosphate-buffered saline pH 7.4) and the simultaneous online detection of Cu, Fe, and Zn in the column effluent by an inductively coupled plasma atomic emission spectrometer (ICP-AES). Whereas most previous studies reported on the analysis of serum, our investigations clearly demonstrated that the analysis of plasma within 30 min of collection results in the detection of one more Cu peak (blood coagulation factor V) than has been previously reported (transcuprein, ceruloplasmin, albumin-bound Cu, and small molecular weight Cu). The average amount of Cu associated with these five proteins corresponded to 21, 18, 21, 30 and 10% of total plasma Cu, respectively. In contrast, only two Fe metalloproteins (ferritin and transferrin, corresponding to an average of 9 and 91% of total plasma Fe) and approximately five Zn metalloproteins (α₂-macroglobulin and albumin-bound Zn, which corresponded to an average of plasma Zn) were detected. Metalloproteins were assigned on the basis of the coelution of the corresponding metal and protein identified by immunoassays or activity-based enzyme assays. The SEC-ICP-AES approach developed allowed the determination of approximately 12 Cu, Fe, and Zn metalloproteins in rabbit plasma within approximately 24 min and can be applied to analyze human plasma, which is potentially useful for diagnosing Cu-, Fe-, and Zn-related diseases. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Parts of the work described in this paper were presented at HPLC 2007 in Ghent, Belgium. An erratum to this article can be found at     

Interfacial electrochemical electron transfer in biology - towards the level of the single molecule

Physical electrochemistry has undergone a remarkable evolution over the last few decades, integrating advanced techniques and theory from solid state and surface physics. Single-crystal electrode surfaces have been a core notion, opening for scanning tunnelling microscopy directly in aqueous electrolyte (in situ STM). Interfacial electrochemistry of metalloproteins is presently going through a similar transition. Electrochemical surfaces with thiol-based promoter molecular monolayers (SAMs) as biomolecular electrochemical environments and the biomolecules themselves have been mapped with unprecedented resolution, opening a new area of single-molecule bioelectrochemistry. We consider first in situ STM of small redox molecules, followed by in situ STM of thiol-based SAMs as molecular views of bioelectrochemical environments. We then address electron transfer metalloproteins, and multi-centre metalloenzymes including applied single-biomolecular perspectives based on metalloprotein/metallic nanoparticle hybrids.     

Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS

Metals are essential for protein function as cofactors to catalyze chemical reactions. Disruption of metal homeostasis is implicated in a number of diseases including Alzheimer''s and Parkinson''s disease, but the exact role these metals play is yet to be fully elucidated. Identification of metalloproteins encounters many challenges and difficulties. Here we report an approach that allows metalloproteins in complex samples to be quantified. This is achieved using size exclusion chromatography coupled with inductively coupled plasma - mass spectrometry (SEC-ICP-MS). Using six known metalloproteins, the size exclusion column can be calibrated and the respective trace elements (iron, copper, zinc, cobalt, iodine) can be used for quantification. SEC-ICP-MS traces of human brain and plasma are presented. The use of these metalloprotein standards provides the means to quantitatively compare metalloprotein abundances between biological samples. This technique is poised to help shed light on the role of metalloproteins in neurodegenerative disease as well as other diseases where imbalances in trace elements are implicated.     

The influence of electrical stimulation of vagus nerve on elemental composition of dopamine related brain structures in rats

Recent studies of Parkinson's disease indicate that dorsal motor nucleus of nerve vagus is one of the earliest brain areas affected by alpha-synuclein and Lewy bodies pathology. The influence of electrical stimulation of vagus nerve on elemental composition of dopamine related brain structures in rats is investigated. Synchrotron radiation based X-ray fluorescence was applied to the elemental micro-imaging and quantification in thin tissue sections. It was found that elements such as P, S, Cl, K, Ca, Fe, Cu, Zn, Se, Br and Rb are present in motor cortex, corpus striatum, nucleus accumbens, substantia nigra, ventral tectal area, and dorsal motor nucleus of vagus. The topographic analysis shows that macro-elements like P, S, Cl and K are highly concentrated within the fiber bundles of corpus striatum. In contrast the levels of trace elements like Fe and Zn are the lowest in these structures. It was found that statistically significant differences between the animals with electrical stimulation of vagus nerve and the control are observed in the left side of corpus striatum for P (p = 0.04), S (p = 0.02), Cl (p = 0.05), K (p = 0.02), Fe (p = 0.04) and Zn (p = 0.02). The mass fractions of these elements are increased in the group for which the electrical stimulation of vagus nerve was performed. Moreover, the contents of Ca (p = 0.02), Zn (p = 0.07) and Rb (p = 0.04) in substantia nigra of right hemisphere are found to be significantly lower in the group with stimulation of vagus nerve than in the control rats.     

An improved method for estimating original mineral contents in excavated bone using sulfur

Trace element analysis in excavated bones is complicated by the lack of a reliable index for estimating the original amount of bone material. In this study, we subjected modern human bones to alkali treatment to simulate aging. Alkali treatment of vertebrae with attached muscle did not affect sulfur (S) content; it increased the magnesium (Mg), phosphorus (P), and zinc (Zn) contents, and tended to decrease iron (Fe) content of the bones. When vertebrae cleaned of muscle were used, alkali treatment did not affect S and Fe contents but increased Mg, P, Ca, and Zn contents Ca and S contents were higher in excavated bones (200–1300 yr old) than in their surrounding soils. In contrast, S, Mg, and Ca contents per dry weight did not differ between the excavated bones and the alkali-treated modern bones. These results indicate that S can provide a more accurate index of excavated bones than the often-used Ca content or dry wt measures, especially for bones excavated from calcium-rich soils.     

Elemental composition of subcellular structures of human spermatozoa. A study by energy dispersive analysis of X-ray

The elemental chemical composition of selected intracellular structures (membrane, mitochondria and nucleus) of the human spermatozoa were studied by the combined use of energy dispersive analyses of X-rays and electron microscopy and the results compared with those obtained by atomic absorption spectrometry of isolated subcellular structures (heads and tails) of the same type of cells. In nuclei EDAX studies showed the following relative concentrations P>S>Mg, K, Zn, Si>Ca, Fe. The mitochondrial spectrum showed the presence of important concentrations of Ca>Fe, K, P>Mg, S>Mn. Human spermatozoa membrane was found to be particularly rich in Ca, S and $\text{Z}$n. By atomic absorption it was found that K was the most concentrated element in both isolated fractions (heads and tails). Sperm heads were found richer than tails in Na, Cu and Zn, while sperm tails had higher concentrations of Ca. The zinc concentration of human sperm cells and their subfractions was considerably higher than the reported Zn concentration in any other human cells or their subfractions.     

应用电子探针对植物根际和根内营养元素微区分布的探讨

用电子探针可检测出玉米、大豆根际和根内含有Na,Mg,Al,Si,P,S,Cl,K,Ca,Ti,Fe,Cu和Zn 13种元素。这些元素在根际土壤、粘液层和根组织内的含量分布有一定的规律性。除Si,Al,Ca,Fe在根际土壤中峰值较高外,Ti仅在土壤中达到可检测量;S,Fe和Zn富集在粘液层,Mg,P,Cl只在根组织内才有较明显的峰。这些规律可作为区分根—土界面的参考指标。K含量在根内明显高于根际土壤,并由表皮层到中柱径向增加;Ca则与K不同,且受植物种类的影响。     

Probing the viral metallome: searching for metalloproteins in bacteriophage λ-- the hunt begins

Although the proteome and genome of bacteriophages are well developed, there is little knowledge about metals and their interactions with the phages, even though metals have been observed in stabilizing phage particles. With expanding studies of phage display and its promising applications, metalloprotein investigations in the bacteriophage areas are necessary to understand whether or not metalloproteins are included in the viral coat proteome. Since these virus studies are still in their infancy, lambda phage was chosen due to its high metal-binding potential as suggested by the cysteine/methionine rich proteins in the viral coat. After large-scale preparation and further purification of lambda phage according to standard protocols, state-of-the-art metallomics techniques via combinations of chromatographies and mass spectrometries were utilized for screening metal-associated species in lambda phage. The lambda phage sample was first separated using non-denaturing size exclusion chromatography with selective metal detection by ICPMS for screening associated metals and generating size distribution fractions for the various metal species, some of which include metalloproteins. Various molecular size distribution patterns were exhibited for the metals detected, Mn, Fe, Co, Ni, Cu and Zn, at different molecular weight ranges. On the other hand numerous other metals were not associated with the coat proteins, as they were not detected in the different molecular weight fractions. Further identification for putative metallopeptides and metalloproteins was accomplished by collecting various metal species' fractions offline and subsequently analyzing tryptically-digested fractions via nanoLC-Chip-ESI-MS. By searching appropriate MS databases with both Spectrum Mill and MASCOT search engines, the main capsid protein, gpE, a capsid decoration protein, gpD, and main tail component protein, gpV, were found and are known for associations with the detected transition metals. These findings will likely provide valuable information for lambda phage engineered applications.     

Effects of three chelating agents,EDTA, NTA,and TPP,on the concentration of elements in rat tissues

Ethylene diamine tetraacetic acid (EDTA), nitrilotriacetic acid (NTA), and tripolyphosphate (TPP) sodium salts were given orally to rats at the dose of 1 mmol/kg/d for 35 d. The concentrations of Na, K, Ca, Mg, P, S, Fe, Sr, Cu, and Zn were determined in blood, plasma, brain, heart, muscle, liver, kidney, duodenum, and bone of control rats and of the rats receiving EDTA, NTA, and TPP. The main effect induced by EDTA, NTA, and TPP was a decrease of the concentrations of several elements Ca, Mg, Fe, P in the duodenum. Otherwise, EDTA induced an increase of Zn in the kidney (+ 20%), NTA, an increase of Fe in liver (+ 29%), and particularly an increase of Zn in bone (+ 44%). TPP induced a slight decrease of Zn and Cu in liver. In conclusion, EDTA, NTA, and TPP taken orally at the dose of 1 mmol/kg/d for 35 d induced moderate changes of the concentrations of some elements in rat tissues, but without signs of toxicity.     

Effects of thyroparathyroidectomy and of thyroxin and calcitonin on the tissue distribution of twelve elements in the rat

The effects of thyroparathyroidectomy (TPTY) and of replacement therapy using thyroxin (T4) and calcitonin (CT) on the tissue distribution of elements were studied in the rat under semichronic conditions. The elements Na, K, Ca, Mg, Fe, S, P, Rb, Sr, Mn, Cu, and Zn were determined in whole blood, plasma, brain, liver, heart, kidney, skeletal muscle, and bone. TPTY modified concentrations of all elements tested but only small changes were observed for K, Mg, S, and P. The mineral bone composition was slightly modified, 28 d after TPTY, whereas plasma was the most altered. The consequences of TPTY were corrected fairly well by T4 for Na, Cu, Zn, Fe, and S, and by CT for K, P, Rb but with less efficiency for Ca. This study revealed that hormones of the thyroid gland, mainly T4, play an important role in the plasma and tissue balance of elements. It is suggested that T4 participates in tissue fixation of Cu, Zn, and Fe and that CT influences phosphoremia and cellular Ca binding.     

Synchrotron radiation for direct analysis of metalloproteins on electrophoresis gels

Metalloproteomics requires analytical techniques able to assess and quantify the inorganic species in metalloproteins. The most widely used methods are hyphenated techniques, based on the coupling of a high resolution chromatographic method with a high sensitivity method for metal analysis in solution. An alternative approach is the use of methods for solid sample analysis, combining metalloprotein separation by gel electrophoresis and direct analysis of the gels. Direct methods are based on beam analysis, such as lasers, ion beams or synchrotron radiation beams. The aim of this review article is to present the main features of synchrotron radiation based methods and their applications for metalloprotein analysis directly on electrophoresis gels. Synchrotron radiation X-ray fluorescence has been successfully employed for sensitive metal identification, and X-ray absorption spectroscopy for metal local structure speciation in proteins. Synchrotron based methods will be compared to ion beam and mass spectrometry for direct analysis of metalloproteins in electrophoresis gels.