Calcium carbonate deposition in a cell wall sac formed in mulberry idioblasts

Summary. Although calcium carbonate is known to be a common biomineral in plants, very little attention has been given to the biological control of calcium carbonate deposition. In mulberry leaves, a subcellular structure is involved in mineral deposition and is described here by a variety of cytological techniques. Calcium carbonate was deposited in large, rounded idioblast cells located in the upper epidermal layer of mulberry leaves. Next to the outmost region (“cap”) of young idioblasts, we found that the inner cell wall layer expanded to form a peculiar outgrowth, named cell wall sac in this report. This sac grew and eventually occupied the entire apoplastic space of the idioblast. Inside the mature cell wall sac, various cellulosic membranes developed and became the major site of Ca carbonate deposition. Concentrated Ca²⁺ was pooled in the peripheral zone, where small Ca carbonate globules were present in large numbers. Large globules were tightly packed among multiple membranes in the central zone, especially in compartments formed by cellulosic membranes and in their neighboring membranes. The maximum Ca sink capacity of a single cell wall sac was quantified using enzymatically isolated idioblasts as approximately 48 ng. The newly formed outgrowth in idioblasts is not a pure calcareous body but a complex cell wall structure filled with substantial amounts of Ca carbonate crystals. Correspondence and reprints: Graduate School of Science and Technology, Kyoto Institute of Technology, Goshokaido, Matsugasaki, Sakyo, Kyoto 606-8585, Japan.     

Tolerance,accumulation and distribution of zinc and cadmium in hyperaccumulator Potentilla griffithii

In this study, zinc (Zn) and cadmium (Cd) tolerance, accumulation and distribution was conducted in Potentilla griffithii H., which has been identified as a new Zn hyperaccumulator found in China. Plants were grown hydroponically with different levels of Zn²⁺ (20, 40, 80 and 160 mg L^?1) and Cd²⁺ (5, 10, 20 and 40 mg L^?1) for 60 days. All plants grew healthy and attained more biomass than the control, except 40 mg L^?1 Cd treatment. Zn or Cd concentration in plants increased steadily with the increasing addition of Zn or Cd in solution. The maximum metal concentrations in roots, petioles and leaves were 14,060, 19,600 and 11,400 mg kg^?1 Zn dry weight (DW) at 160 mg L^?1 Zn treatment, and 9098, 3077 and 852 mg kg^?1 Cd DW at 40 mg L^?1 Cd treatment, respectively. These results suggest that P. griffithii has a high ability to tolerate and accumulate Cd and Zn, and it can be considered not only as Zn but also as a potential cadmium hyperaccumulator. Light microscope (LM) with histochemical method, scanning electron microscope combined with energy dispersive spectrometry (SEM-EDS) and transmission electron microscope (TEM) were used to determine the distribution of Zn and Cd in P. griffithii at tissue and cellular levels. In roots, SEM-EDS confirmed that the highest Zn concentration was found in xylem parenchyma cells and epidermal cells, while for Cd, a gradient was observed with the highest Cd concentration in rhizodermal and cortex cells, followed by central cylinder. LM results showed that Zn and Cd distributed mainly along the walls of epidermis, cortex, endodermis and some xylem parenchyma. In leaves, Zn and Cd shared the similar distribution pattern, and both were mostly accumulated in epidermis and bundle sheath. However, in leaves of 40 mg L^?1 Cd treatment, which caused the phytotoxicity, Cd was also found in the mesophyll cells. The major storage site for Zn and Cd in leaves of P. griffithii was vacuoles, to a lesser extent cell wall or cytosol. The present study demonstrates that the predominant sequestration of Zn and Cd in cell walls of roots and in vacuoles of epidermis and bundle sheath of leaves may play a major role in strong tolerance and hyperaccumulation of Zn and Cd in P. griffithii.     

Response of Vicia faba L. to metal toxicity on mine tailing substrate: Geochemical and morphological changes in leaf and root

Vicia faba L. seeds were grown in a pot experiment on soil, mine tailings, and a mixture of both to mimic field situations in cultivated contaminated areas near mining sites. Metals in the substrates and their translocation in root, stem and leaf tissues were investigated, including morphological responses of plants growing on mine tailings. Metal concentration – and generally bioaccumulation – was in the order: roots > leaves > stems, except Pb and Cd. Translocation was most significant for Zn and Cd, but limited for Pb. Metal concentration in root and leaf was not proportional to that in the substrates, unexpectedly the minimum being observed in the mixed substrate whilst plant growth was retarded by 20% (38% on tailings). Calcium, pH, organic matter and phosphorus were the main influencing factors for metal translocation. The ultrastructure of V. faba L. cells changed a lot in the mine tailings group: root cell walls were thickened with electron dense Pb, Zn and C particles; in chloroplasts, the number of plastoglobuli increased, whereas the thylakoids were swollen and their number decreased in grana. Finally, needle-shaped crystalline concretions made of Ca and P, with Zn content, were formed in the apoplast of the plants. The stratagems of V. faba L. undergoing high concentrations of toxic metals in carbonate substrate, suggest root cell wall thickening to decrease uptake of toxic metals, a possible control of metal transport from roots to leaves by synthesizing phytochelators–toxic metal complexes, and finally a control of exceeded Ca and metal concentration in leaves by crystal P formation as ultimate response to stress defense. The geochemical factors influencing metal availability, guaranty a reduction of metal content in plant growing on mixed tailing/soil substrate as far as carbonate are not completely dissolved.     

The concentrations of major and trace elements in rat kidney: Aging effects and mutual relationships

The concentrations of 26 major to trace elements in rat kidneys aging from 5 to 113 weeks old were determined. The rats investigated were the same rats used previously reported to have 29 elements in bones (femurs). The samples were decomposed by high purity nitric acid and hydrogen peroxide. Eight elements (Na, Mg, Si, P, K, Ca, Fe and Zn) were determined using inductively coupled plasma atomic emission spectrometry (ICP-AES) and 18 elements (Mn, Co, Ni, Cu, As, Se, Rb, Sr, Mo, Cd, Sn, Sb, Cs, Ba, Tl, Pb, Bi and U) were determined using inductively coupled plasma mass spectrometry (ICP-MS). The aging effects on the concentrations of these elements and mutual elemental relationships were investigated. Analysis of variance (ANOVA) for age variations indicated that the concentrations of P, K, Mn and Mo were almost constant across the age of rats (p > 0.3). The concentration of many elements such as Na, Mg, Ca, Fe, Co, Cu, Zn, As, Se, Cd, Sn, Sb, Tl, Pb and Bi, showed significant increasing trends (p < 0.01) with different patterns. Rubidium, Cs, Pb and Bi showed significant age variations but not monotonic trends. Silicon, Ni, Sr, Ba and U showed large concentration scatterings without any significant trends (p > 0.01). The metabolism of these elements may not be well established in the kidney. Many toxic elements such as As, Cd, Sn, Pb and Bi showed a narrow concentration range among age-matched rats. The kidney may have established metabolic mechanisms to confine or accumulate these toxic elements even though their concentrations are very low (e.g., 10 ng g^?1 of Cd). These elements also closely coupled with Fe. A cluster analysis was performed using an elemental correlation matrix and indicated that these elements, including Fe, formed a cluster. However, another cluster analysis using “an aging effect eliminated” elemental correlation showed different clustering in which the Fe, Cd cluster disappeared.     

Effects of mulberry leaves to replace rapeseed meal on performance of sheep feeding on ammoniated rice straw diet

This study was conducted to investigate effects of leaves of mulberry tree (Morus alba) as a protein supplement to isonitrogenously replace rapeseed meal (RSM) on performance of growing lambs offered ammoniated rice straw (ABRS) (Trial 1), and to evaluate the digestive characteristics of the ABRS supplemented with different ratios of RSM and mulberry leaves in terms of in vitro gas production (Trial 2). In Trial 1, 45 Huzhou lambs were divided into five equal groups according to their body weight and gender. Lambs in each group were kept in three pens (male, female and mixed (one male and two females)), and received one of the following dietary treatments: 100 g RSM (A), 75 g RSM plus 60 g mulberry leaves (B), 50 g RSM plus 120 g mulberry leaves (C), 25 g RSM plus 180 g mulberry leaves (D), and 240 g mulberry leaves (E). All animals were given ABRS ad libitum along with 100 g ground corn per head per day. The intake of ABRS was slightly increased with the supplementary level of mulberry leaves, and hence total intake increased with the increasing level of mulberry leaves. The growth rates were higher in diets A and E than those in other treatments (P<0.05), with little difference between diets A and E, and the slowest in C. Animals of all genders showed a similar trend, though male lambs was higher in weight gain than the female. While feed efficiency was higher in diet A, concentrate consumption per kilogram of weight gain was lower when higher level of mulberry leaves was supplemented (diets D and E). Feed cost per kilogram gain was lower in diets E and A compared to other treatments. Degradation of dry matter in the rumen of sheep were higher for mulberry leaves than for RSM, but crude protein was less degraded for mulberry leaves than for RSM. The potential GP was significantly higher in diet A than those in B, C and D (P<0.05), and higher in E than in C (P<0.05) (Trial 2), indicating a negative associate effect of mulberry leaves and RSM on digestion. It is inferred that mulberry leaves may be used as a protein supplement to ammoniated straw diets to fully substitute for RSM, but these two supplements should unlikely be supplemented together to avoid the negative associate effect.     

Lead,zinc, cadmium hyperaccumulation and growth stimulation in Arabis paniculata Franch

Metal hyperaccumulation is of great interest in recent years because of its potential application for phytoremediation of heavy metal contaminated soils. In this study, a field survey and a hydroponic experiment were conducted to study the accumulation characteristics of lead (Pb), zinc (Zn) and cadmium (Cd) in Arabis paniculata Franch., which was found in Yunnan Province, China. The field survey showed that the wild population of A. paniculata was hyper-tolerant to extremely high concentrations of Pb, Zn and Cd, and could accumulate in shoots an average level of 2300 mg kg^?1 dry weight (DW) Pb, 20,800 mg kg^?1 Zn and 434 mg kg^?1 Cd, with their translocation factors (TFs) all above one. Under the hydroponic culture, stimulatory effects of Pb, Zn and Cd on shoot dry biomass were noted from 24 to 193 μM Pb, 9 to 178 μM Cd and all Zn supply levels in nutrient solution, while the effects were not obvious in the roots. Chlorophyll concentrations in Pb, Zn and Cd treatments showed an inverted U-shaped pattern, consistent with the change of plant biomass. Pb, Zn and Cd concentrations in the shoots and roots increased sharply with increasing Pb, Zn and Cd supply levels. They reached > 1000 mg kg^?1 Pb, 10,000 mg kg^?1 Zn and 100 mg kg^?1 Cd DW in the 24 μM Pb, 1223 μM Zn and 9 μM Cd treatment, respectively, in which the plants grew healthy and did not show any symptoms of phytotoxicity. The TFs of Zn were basically higher than one and the amount of Zn taken by shoots ranged from 78.7 to 90.4% of the total Zn. However, the TFs of Pb and Cd were well below one, and 55.0–67.5% of total Pb and 57.8–83.5% of total Cd was accumulated in the shoots. These results indicate that A. paniculata has a strong ability to tolerate and hyperaccumulate Pb, Zn and Cd. Meanwhile, suitable levels of Pb, Zn and Cd could stimulate the biomass production and chlorophyll concentrations of A. paniculata. Thus, it provides a new plant material for understanding the mechanisms of stimulatory effect and co-hyperaccumulation of multiple heavy metals.     

Accumulation of zinc and cadmium and localization of zinc in Picris divaricata Vant.

Picris divaricata Vant., a plant species native to subtropical China, was recently identified as the first Cd/Zn hyperaccumulator from Asteraceae. P. divaricata was grown from wild collected seed for 4 months in a series of pH adjusted test soils with added Zn levels 0–7000 mg kg⁻¹ and Cd levels 0–150 mg kg⁻¹. Plants did not hyperaccumulate Zn (threshold >3000 μg g⁻¹) and weakly hyperaccumulated Cd with little or no dose–response.P. divaricata has multicellular simple trichomes concentrated on the leaf margins and midrib. X-ray analysis showed that Zn was concentrated in larger trichomes and epidermal cells adjacent to the trichome but virtually absent in other leaf tissues. Within the trichomes, Zn was localized in ovate spots around the tips of individual cells. These tips and other locations in the trichome cell contained black electron dense material when examined with transmission electron microscopy, some of which was identified as SiO₂. Silicon and Mn were concentrated in the same areas as Zn. Si has been previously associated with alleviating Zn, Mn and Cd toxicity. Our results support this observation and further investigation is warranted.Calcium and P were concentrated in the distal tips of trichomes, similar to patterns previously observed for calcicole plants grown in elevated Ca soils. Overall, nonsecretory trichomes from many plant families may have a common origin as tissues adapted to handle a variety of environmental metals.     

The effect of age and gender on 38 chemical element contents in human iliac crest investigated by instrumental neutron activation analysis

ProjectTo understand the role of major, minor, and trace elements in the etiology of bone diseases including osteoporosis, it is necessary to determine the normal levels and age-related changes of bone chemical elements.ProcedureThe effect of age and gender on 38 chemical element contents in intact iliac crest of 84 apparently healthy 15–55 years old women (n=38) and men (n=46) was investigated by neutron activation analysis.ResultsMean values (M±SEM) for mass fraction (on dry weight basis) of Ca, Cl, Co, Fe, K, Mg, Mn, Na, P, Rb, Sr, and Zn for both female and male taken together were Ca – 169±3 g/kg, Cl – 1490±43 mg/kg, Co – 0.0073±0.0024 mg/kg, Fe – 177±24 mg/kg, K – 1820±79 mg/kg, Mg – 1840±48 mg/kg, Mn – 0.316±0.013 mg/kg, Na – 4970±87 mg/kg, P – 79.7±1.5 g/kg, Rb – 1.89±0.22 mg/kg, Sr – 312±15 mg/kg, and Zn – 65.9±3.4 mg/kg, respectively. The upper limit of mean contents of Cs, Eu, Hg, Sb, Sc, and Se were Cs≤0.09 mg/kg, Eu≤0.005 mg/kg, Hg≤0.005 mg/kg, Sb≤0.004 mg/kg, Sc≤0.001 mg/kg, and Se≤0.1 mg/kg, respectively. In all bone samples the contents of Ag, As, Au, Ba, Br, Cd, Ce, Cr, Gd, Hf, La, Lu, Nd, Sm, Ta, Tb, Th, U, Yb, and Zr were under detection limits.ConclusionsThe Ca, Mg, and P contents decrease with age, regardless of gender. Higher Ca, Mg, P, and Sr mass fractions as well as lower Fe content are typical of female iliac crest as compared to those in male bone.     

Changes in the elemental content of rat heart as a result of the fixation in formalin analyzed by synchrotron radiation X-ray fluorescent analysis

The aim of the study was to investigate possible changes in the elemental content of small biological tissue samples (with low weight) during the short period of fixation in formalin. Therefore, the effect of the fixation time (4 h to 6 days) was studied on the elemental content of the tissue. Synchrotron radiation X-ray fluorescent analysis (SRXRF) technique was used for the determination of the elemental concentrations. The levels of Cl, K, Ca, Br, Rb, and Sr in the samples fixed for 4 h (and longer) show significant differences in comparison with levels in fresh samples, because these ions have a high mobility and capacity for moving through the cell membrane. The content of Zn, Se, Fe, and Cu in the samples fixed by formalin solution during 4 h do not show significant differences compared with fresh samples. The ions of these elements can form complexes with the large protein molecules in the tissue and can hardly be removed from it.     

Subcellular distribution and chemical forms of cadmium in Bechmeria nivea (L.) Gaud.

Bechmeria nivea (L.) Gaud. (Ramie) is a promising species for Cd phytoextraction with large biomass and fast growth rate. Nevertheless, little information is available on its tolerance mechanisms towards Cd. Determination of Cd distribution and chemical speciation in ramie is essential for understanding the mechanisms involved in Cd accumulation, transportation and detoxification. In the present study, ramie plants were grown in hydroponics with increasing Cd concentrations (0, 1, 3, 7 mg l^?1). The subcellular distribution and chemical forms of Cd in different tissues were determined after 20 days exposure to this metal. To assess the effect of Cd uptake on plant performance, nitrate reductase activity in leaves and root activity were analyzed during the entire experimental period. Increased Cd level in the medium caused a proportional increase in Cd uptake, and the highest Cd concentration occurred in roots, followed by stems and leaves. Subcellular fractionation of Cd-containing tissues indicated that about 48.2–61.9% of the element was localized in cell walls and 30.2–38.1% in soluble fraction, and the lowest in cellular organelles. Cd taken up by ramie rapidly equilibrated among different chemical forms. Results showed that the greatest amount of Cd was found in the extraction of 1 M NaCl and 2% HAC, and the least in residues in all test tissues. In roots, the subdominant amount of Cd was extracted by d-H₂O and 80% ethanol, followed by 0.6 M HCl. While in stems and leaves, the amount of 0.6 M HCl-extractable Cd was comparable with that extracted by 80% ethanol or d-H₂O. 1 mg l^?1 Cd stimulated nitrate reductase activity in leaves and root activity, while a concentration-dependent inhibitory effect was observed with increasing Cd concentration, particularly at 7 mg l^?1 Cd. It could be suggested that the protective mechanisms evolved by ramie play an important role in Cd detoxification at relatively low Cd concentrations (below 3 mg l^?1 Cd) but become restricted to maintain internal homeostasis with higher Cd stress.     

Copper and zinc in Elodea canadensis from rivers with various pollution levels

The anthropogenic impact of xenobiotics contributes to environmental risk for the aquatic environment and thus, must be controlled. Elodea canadensis, a cosmopolitan aquatic macrophyte with an important role in the ecology of many littoral zones, may provide an integrated record of pollution. Therefore, it was interesting to investigate the accumulation of Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in this species and in water and bottom sediments collected from rivers with various levels of contamination. Of these rivers one control and one polluted was selected for the collection of E. canadensis for an experiment to compare the ability of this species to accumulate Cu and Zn. These elements were supplemented at concentrations (mg L⁻¹) of 0.01, 0.02, 0.03, 0.05, 0.08 and 0.14 as CuSO₄·5H₂O, and 0.4, 0.6, 0.9, 1.4, 2.03 and 3.04 as ZnSO₄·7H₂O and in a mixture containing (mg L⁻¹) 0.01Cu + 0.4Zn, 0.02Cu + 0.6Zn, 0.03Cu + 0.9Zn, 0.05Cu + 1.4Zn, 0.08Cu + 2.03Zn and 0.14Cu + 3.04Zn. After the experiment, E. canadensis from the polluted river contained significantly higher Cu and Zn concentrations when applied separately and also significantly higher Cu and Zn concentrations when applied as a mixture compared to the control river. These higher concentrations in E. canadensis from the polluted river were found in all combinations in the experiment. Thus, E. canadensis habituated in polluted sites to the exposure, and long-term influence of elevated metal levels appeared to be better adapted, and it also exhibited a higher increase in biomass than plants from the control river in all the experimental Cu and Zn solutions. Younger leaves of E. canadensis were more resistant to the effects of Cu and Zn than older leaves. Both Cu and Zn negatively affected the cell structure of older leaves, although the influence of Cu on plasma membrane integrity and chloroplast distribution was stronger than that of Zn. The influence of the Cu + Zn mixture on E. canadensis resulted in less pronounced cell disintegration than the influence of Cu added separately.The explanation of differences in the E. canadensis biomass increase and metal concentrations under the binary Cu and Zn impact needs further examination.     

Calcium protection of PS2 complex of Phaseolus coccineus from cadmium toxicity: In vitro study

The action of 10 and 20 mM Ca against harmful Cd effect on PS2 complex isolated from leaves of Phaseolus coccineus L. cv. Pi?kny Ja? was studied. The changes in fast chlorophyll a fluorescence induction kinetics and protein composition of PS2 complex were the symptoms of Cd toxicity and Ca protection of PS2 complex. Calcium applied at 10 mM concentration prevented F₀ reduction caused by the presence of 250–1000 μM Cd in the incubation mixture, but that of (the variable chlorophyll a fluorescence) F_v, F_m, F_v/F₀, and F_v/F_m only at 250 μM Cd. Ca concentration doubling in the incubation mixture resulted in complete overcoming the toxicity of 250–1000 μM Cd to F_v and F_m. However, the protection of F_v/F₀ and the photochemical efficiency of PS2 (F_v/F_m) from 1000 μM Cd was only partial even at 20 mM Ca. A protective effect of 10 mM Ca on D1, D2 and 17 kDa proteins was found in PS2 complex exposed to 250 μM Cd, and on 43 kDa protein in the complex treated with 500 μM Cd. However, 20 mM Ca counteracted the toxicity of 500 μM Cd to the 43, 47 and 17 kDa proteins, as well as the harmful effect of 1000 μM Cd on 47 and 17 kDa ones.     

Heavy metal bioaccumulation by the organs of Phragmites australis (common reed) and their potential use as contamination indicators

The concentrations of heavy metals in the roots, rhizomes, stems and leaves of the aquatic macrophyte Phragmites australis (common reed), and in the corresponding water and sediment samples from the mouth area of the Imera Meridionale River (Sicily, Italy), were investigated to ascertain whether plant organs are characterized by differential accumulation, and to test the suitability of the various organs for heavy metal biomonitoring of water and soil. Heavy metals considered were Cd, Cr, Cu, Hg, Mn, Ni, Pb, Zn. Results showed that belowground organs were the primary areas of metal accumulation. In particular, metal concentrations in plant organs decreased in the order of root > rhizome ≥ leaf > stem. All four organs showed significant differences in concentration for Cr, Hg, Mn, Zn, thus suggesting low mobility from roots to rhizomes and to aboveground organs. Although the organs followed different decreasing trends of metal concentration, the trend Mn > Zn > Pb > Cu was found in each plant organ. Mn showed the highest concentrations in all organs whereas the lowest concentrations regarded Cd and Cr in the belowground and aboveground organs, respectively. The toxic threshold was exceeded by Cr in roots, rhizomes and leaves, Mn in roots and leaves, Ni in roots. The highest average concentrations were found as follows: Cd, Hg, Pb, Zn in root, Cr, Mn, Ni in sediment, Cu in water. Positive linear relationships were found between heavy metal concentrations in all plant organs and those in water and sediment, thus indicating the potential use of such organs for pollution monitoring of water and sediment. Advantages of using plant species as biomonitors, especially Phragmites australis, were also discussed.     

Age-related effects of major and trace element concentrations in rat liver and their mutual relationships

The concentrations of 22 major and trace elements in livers from rats aging from 5 to 113 weeks old were determined. The rats investigated were the same rats previously reported with respect to 29 elements in bones (femur) and 26 elements in kidneys. The samples were decomposed with high-purity nitric acid and hydrogen peroxide. Seven elements (Na, Mg, P, K, Ca, Fe and Zn) were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES), and 15 elements (Mn, Co, Cu, As, Se, Rb, Sr, Mo, Cd, Sn, Sb, Cs, Ba, Pb and Bi) were determined by inductively coupled plasma mass spectrometry (ICP-MS). Analysis of variance (ANOVA) for age variations indicated that the concentrations of many elements, such as Mg, P, K, Mn, Fe, Cu, Zn, Sr, Mo and Cd, were almost constant across the ages of the rats with the exception of 5 weeks old (p > 0.05). Arsenic, Pb and Bi showed significant increasing trends, while Na and Co showed decreasing trends (p < 0.01). Selenium showed a decreasing trend except at the initial stage of 5–9 weeks old. Calcium, Rb, Sn, Sb, Cs and Ba showed significant age-related variations, but their patterns were not monotonic. The liver clearly contrasts with the kidneys, in which many elements showed significant age-related variations with increasing trends. The concentration ranges of Mg, P, K, Mn, Cu, Zn, and Mo were controlled within 15% across all ages of rats. The homeostasis of the aforementioned elements may be well established in the liver. The toxic elements, such as Cd, Pb and Bi, showed a narrow concentration range among age-matched rats.     

Anatomical variations in primary teeth microelements with known differences in lead content by micro-Synchrotron Radiation X-Ray Fluorescence (μ-SRXRF) – A preliminary study

Shed teeth have been proposed as trace element biomarkers. This study determined variations in the spatial distribution of Ca, K, Zn, Pb, Mn, Cu, and Sr in four anatomical locations: superficial enamel (SE, 0–10 μm), subsuperficial enamel (SSE, 10–30 μm), primary dentin (PD), and secondary dentin (SD). Five primary incisors were analyzed by micro Synchrotron Radiation X-Ray Fluorescence (μ-SRXRF). Two teeth had low concentrations of lead in the SE (<250 μg/g), while three contained very high lead concentrations in the SE (>2000 μg/g). Teeth were sliced, and five spot measurements (20 μm beam diameter) were accomplished in each location. The data are shown as absolute values and as the ratio between the different elements and Ca. The distribution of K was close to that of Ca. Zn was the third most abundant element, with the highest levels being found in the SE and SD and low levels detected in the PD. Increasing Sr levels were found progressing from the enamel to the dentin, with the highest levels being found in the SD, a distribution that was unique. Pb, Mn, and Cu exhibited a similar trend, with higher signals for these elements detected in the SE. This study provides preliminary data on the heterogeneous distribution of different elements in the tooth, highlighting the importance of the first 10 μm of the SE for determination of some elements, such as Zn, Pb, Mn, and Cu.     

Histopathology and cytotoxicity as biomarkers in treated rats with cadmium and some therapeutic agents

The present study aimed to investigate the protective role of ascorbic acid (vitamin C) and zinc (Zn) against cadmium (Cd) induced histopathological changes in tissues of liver, kidney, lung and testis of rats as well as chromosomal aberrations. For this purpose, 60 male albino rats were divided into six groups; each group contained 10 animals. The first group served as control and was given only distilled water. The second and third groups received distilled water supplemented with 2 g ascorbic acid/l and 500 mg Zn/l, respectively. The fourth group received a daily oral dose containing 3 mg Cd/kg b.w. (1/30 LD₅₀). The fifth group received Cd + ascorbic acid (3 mg Cd/kg b.w. + 2 g ascorbic acid/l), while the sixth group received Cd + Zn (3 mg Cd/kg b.w. +500 mg Zn/l). The treatment in all groups lasted for 90 consecutive days. Rats exposed to cadmium showed severe histopathological changes in the liver, kidney, lung and testicular tissues as well as chromosomal aberrations such as: break, ring, centromeric separation and polyploidy. Co-treatment with zinc partially improved the histopathological changes and chromosomal aberrations while co-treatment with vitamin C exhibited a more protective role and markedly reduced tissues damage induced by Cd.     

Relationship of strontium and calcium concentrations with the parameters of cell structure in Siberian spruce and fir tree-rings

This study addressed distribution of calcium and strontium in Siberian spruce (Picea obovata Ledeb.) and Siberian fir (Abies sibirica Ledeb.) tree-rings and its dependence on these woody species cell structure. Calcium concentration was found to decrease gradually from earlywood to latewood, whereas strontium showed an opposite trend. However, their trends at the scale of several rings are co-directed in the samples analyzed. A strong linear relationship was identified between the distribution of Sr/Ca concentration ratio and tree-ring density profile for both woody species. Radiographic density of Siberian spruce tree-ring cell walls and Ca and Sr concentrations in them were determined to have negative correlation with cell wall thickness. In earlywood of annual rings of a spruce the radiographic density of cell wall reaches 2.0 g/cm³ and decreases to 1.2 g/cm³ in latewood. The hypothesis put forward in this study to explain these strontium and calcium distributions in the tree-rings is that the concentrations of the element ions change with development of different cell wall layers. The high value of radiographic density of a cellular wall in earlywood and its relationship with cell wall thickness can be explained by the presence of ions of calcium in a cellular wall. Ions of calcium absorb X-ray radiation more strongly in comparison with light chemical elements. It can become the reason of observable relationship between radiographic density of cell wall and cell wall thickness.     

Evidence of century-scale environmental changes: Trace element in tree-ring from Fuling Mausoleum Shenyang,China

In order to detect the response of trees to environmental pollution in Shenyang during the last century, tree rings of two 100+ year old Chinese pine (Pinus tabulaeformis Carr.) were analyzed for nine trace element concentrations (K, Ca, Mg, Mn, Cu, Zn, Al, Pb and Cd) and compared to local environmental change. The concentrations of Cu, Zn and Al had significant (p < 0.01) increasing trends after 1940s and correlated significantly (p < 0.01), with increasing production of nonferrous metal from local industrial processing. Concentrations of nutrient element (K, Ca and Mg) showed significant (p < 0.01) increasing trend from heartwood to sapwood with declines in the last three decades. Significant correlation between climatic factors and element concentration in tree rings were also found, in which the temperature (November to May) and relative humidity in April and May showed significant positive correlation with Ca, Zn and Cu (p < 0.05) as well as Al (p < 0.01) concentrations in tree rings. April rainfall showed significant positive correlation with K and Zn (p < 0.05) as well as Al (p < 0.01) concentration in tree rings. The results suggest that tree rings from Chinese pine in Shenyang are sensitive to environmental change and have the potential to be used as a biomonitor for the environmental pollution. The results also demonstrate the feasibility of applying dendrochemical techniques to areas with limited samples.     

Effects of zinc pre-treatment on blood glutathione,serum zinc and kidney histological organisation in male rats exposed to cadmium

The effects of sub-chronic exposure to cadmium (Cd) on the blood glutathione, serum zinc and on the kidney histological organisation in rats as well as the possible protective role of zinc (Zn) are the object of this study. For this purpose, 60 male Wistar rats (8 weeks old) were divided into three groups: the first group was exposed to Cd in the form of CdCl₂, administered in five doses (each of 0.4 mg Cd/kg b.w.) on days 5, 10, 15, 20 and 25, giving a total dose of 2 mg Cd/kg b.w., i.p.; the second group was simultaneously exposed to Zn and Cd with the same timeline and the same doses of Cd as the first group but with, in addition, injections of Zn in the form of ZnCl₂, administered in doses of 0.8 mg Zn/kg b.w., giving a total dose of 4 mg Zn/kg b w, i.p.; a control group received 0.5 mL of physiological saline in an identical manner. Intoxication with Cd was followed by a significant decrease in blood glutathione, increase in oxidized glutathione as well as histological damage in kidneys. Pre-treatment with Zn exhibited a protective role against Cd toxicity with a significant decrease in serum zinc content. This fact may be explained by an excessive use of zinc in metallothionein synthesis as a cadmium detoxification agent.