Living Target of Ce(III) Action on Horseradish Cells: Proteins on/in Cell Membrane

Positive and negative effects of rare earth elements (REEs) in life have been reported in many papers, but the cellular mechanisms have not been answered, especially the action sites of REEs on plasma membrane are unknown. Proteins on/in the plasma membrane perform main functions of the plasma membrane. Cerium (Ce) is the richest REEs in crust. Thus, the interaction between Ce(III) and the proteins on/in the plasma membrane, the morphology of protoplast, and the contents of nutrient elements in protoplast of horseradish were investigated using the optimized combination of the fluorescence microscopy, fluorescence spectroscopy, circular dichroism, scanning electron microscopy, and X-ray energy dispersive spectroscopy. It was found that Ce(III) at the low concentrations (10, 30???M) could interact with proteins on/in the plasma membrane of horseradish, leading to the improvement in the structure of membrane proteins and the plasma membrane, which accelerated the intra-/extra-cellular substance exchange and further promoted the development of cells. When horseradish was treated with Ce(III) at the high concentrations (60, 80???M), Ce(III) also could interact with the proteins on/in the plasma membrane of horseradish, leading to the destruction in the structure of membrane proteins and the plasma membrane. These effects decelerated the intra-/extra-cellular substance exchange and further inhibited the development of cells. Thus, the interaction between Ce(III) and proteins on/in the plasma membrane in plants was an important reason of the positive and negative effects of Ce(III) on plants. The results would provide some references for understanding the cellular effect mechanisms of REEs on plants.     

Microcalorimetric study of the effect of Ce(III) on metabolic activity of mitochondria isolated from indice rice 9311

Rare earth elements (REEs) have beneficial influence on plant growth and are widely used in agriculture practice, but little is known about behavior of the REEs on mitochondria in plant cells. Thermogenic metabolic curves were determined by the ampoule method at 303 K using a TAM air isothermal microcalorimeter in mitochondria isolated from indice rice 9311 (Oryza sativa L.), and the effect of Ce(III) on mitochondrial metabolism was investigated. By analyzing the obtained heat flux curves, the crucial parameters such as activity recovery rate constant (k) and maximum heat power (P(m)) were investigated. Application of Ce3+ in concentrations ranging from 0 to 120 microg/ml significantly increased k and P(m) values, with the maximum reaching 261 and 180% of the control, respectively. Concentrations from 140 to 150 microg/ml had the opposite effect. These results were consistent with previous reports on the effects of REEs on plant growth. It was concluded that the Ce(III)-induced change of mitochondrial metabolic activity is a possible mechanism by which Ce(III) influenced indice rice 9311 growth.     

Electron Microscopic Autoradiography: An Improved Procedure Using Aclar® Film and Grid-Sticks

A simple, time saving technique for electron microscopic autoradiography (EMARG) has been developed using both Aclar® film and Grid-sticks. Rats were given 3 mg/kg [pyridine-5-³H] nicorandil orally. After 15 min, animals were sacrificed, the hearts were removed, and myocardial samples for EMARG were prepared. Silver grains were detected clearly in myofibrils and in mitochondria, all showing well preserved ultrastructure. The use of Grid-sticks in the procedure improves handling of bulk samples.     

Behavior of neutron-activated uranium dioxide dust particles in the gastrointestinal tract of the rat

The behavior of neutron-irradiated, simulated Chernobyl UO2 particles containing 141Ce, 144Ce, 95Zr, 95Nb, and 103Ru in the gastrointestinal tract was investigated to obtain basic information for dosimetric and risk analyses of nuclear accidents. After the UO2 particles were administered to rats intragastrically, the distribution and retention of specific radionuclides were studied by using whole-body autoradiography and gamma-spectrometric analysis of tissues. None of the radionuclides were detected in liver, kidney, muscle, bone, brain, blood, and urine. Approximately 98% of the total administered radioactivity was excreted in feces within 3 days. A two times greater intestinal retention (about 6%) of 95Nb than for the other radionuclides was observed 1 day after administration. The results indicate that this kind of relatively insoluble particulate material is not absorbed or retained significantly in the epithelial cells of the intestinal wall. Fallout particles containing high-energy beta sources, 106Ru and 144Ce, result in a very high radiation dose (up to several Gy/day) in the vicinity of a hot particle. Niobium-95 with low average beta energy (0.043 MeV (100%)) does not increase the total dose to the GI tract significantly despite its longer retention in the intestine. Evaluation of the biological effects of these particles in the GI tract by using a dosimetric model based on uniform distribution of activity may be misleading.     

稀土元素在赣南非稀土矿区和不同稀土矿区土壤-铁芒萁(Dicranopteris linearis)系统中的分布、累积和迁移

ICP-M法测定了江西赣南地区非稀土矿区和4处不同稀土矿区内,土壤-铁芒萁系统中15个稀土元素的含量,并对稀土元素在土壤剖面层及铁芒萁植物体内的分布、迁移特征进行了研究.结果表明,稀土元素总量在土壤剖面层的底土层含量最高,但表土层铈相对富集.稀土元素在铁芒萁植物体内的分布规律是叶、根＞茎＞叶柄.铁芒萁根中稀土元素的丰度与其母土表土层,尤其是母土表土层可溶态稀土元素的分布模式基本相似.稀土元素在铁芒萁体内的迁移过程中,发生了明显的分馏作用,茎、叶柄、叶中的重稀土相对贫乏.     

感耦等离子体发射光谱法研究土壤—铁芒萁系统中稀土元素的分布、累积和迁移特征

用感耦等离子体发射光谱法（ICP－AES）测定了东西赣南地区非稀土矿区和4处不同稀土矿区内,土壤－铁芒萁系统中La、Ce、Pr、Nd、Sm、Gd、Dy、Yb和Y的含量,并对其在土壤剖面层及铁芒萁植物体内的分布、迁移特征进行了研究,结果表明,土训层的底土层含量最高,但表土层铈相对富集、稀土元素总量（∑REE）在铁芒萁植物体内的分布规律是叶＞根＞茎＞叶柄,单一稀土的分布规律各异,La、Ce、Pr、Nd的分布规律表现为：叶＞根＞茎＞叶柄;Sm和Gd在不同采样点表现为叶＞根＞茎＞叶柄或根＞叶＞茎＞叶柄;Dy、Yb和Y均有3种不同的分布模式：叶＞根＞茎＞叶柄、根＞叶＞茎＞叶柄及根＞茎＞叶＞叶柄,稀土元素在铁芒萁体内的迁移过程中,发生了明显的分馏作用,茎、叶柄、叶中的重稀土相对贫乏。     

感耦等离子体发射光谱法研究土壤-铁芒萁系统中稀土元素的分布、累积和迁移特征

用感耦等离子体发射光谱法(ICP-AES)测定了江西赣南地区非稀土矿区和处不同稀土矿区内,土壤-铁芒萁系统中La、Ce、Pr、Nd、Sm、Gy、Db、Yb和Y的含量,并对其在土壤剖面层及铁芒萁植物体内的分布、迁移特征进行了研究.结果表明,土壤剖面层的底土层含量最高,但表土层铈相对富集.稀土元素总量(∑REE)在铁芒萁植物体内的分布规律是叶＞根＞茎＞叶柄,单一稀土元素的分布规律各异,La、Ce、Pr、Nd的分布规律表现为:叶＞根＞茎＞叶柄;Sm和Gd在不同采样点表现为叶＞根＞茎＞叶柄或根＞叶＞茎＞叶柄;Dy、Yb和Y均有3种不同的分布模式:叶＞根＞茎＞叶柄、根＞叶＞茎＞叶柄及根＞茎＞叶＞叶柄.稀土元素在铁芒萁体内的迁移过程中,发生了明显的分馏作用,茎、叶柄、叶中的重稀土相对贫乏.     

长期喷施稀土对土壤-植物(小麦)系统中稀土元素分布、累积及运移的影响

在我国施用稀土时间最长的黑龙江省花园农场,研究比较了施用稀土12 a和不施稀土的对照处理上小麦和土壤中的稀土元素的分布、累积及运移.结果表明,长期叶面喷施稀土并未造成耕层土壤和下部土层的稀土元素的累积.成熟期小麦植株各部位的稀土元素含量顺序为根>叶>茎、壳;稀土元素主要累积在根部,其次是叶,茎和壳累积较少.喷施处理小麦根部的稀土元素累积量高于对照,叶部也有此趋势,而茎、壳等部位差异不大.根、茎、叶、壳稀土元素分布模式与土壤中相似,与施用的常乐稀土差别较大;长期喷施稀土未曾造成籽粒中稀土元素的明显累积.     

Ultrastructural studies of the intracellular translocation of endocytosed alpha-foetoprotein (AFP) by cytochemistry and of the uptake of 3H-arachidonic acid bound to AFP by autoradiography in rat rhabdomyosarcoma cells

A covalent conjugate of alpha-foetoprotein (AFP) and horseradish peroxidase (HRP) has been used to follow, at the ultrastructural level, the pathway of AFP uptake and translocation in a rat rhabdomyosarcoma cell line. The cells were incubated for several times at 4 degrees C and/or 37 degrees C, and fixed. AFP-HRP was found to enter the cells via coated pits and receptosomes and to move to tubular elements of the trans-reticular portion of the Golgi. Some observations suggest that AFP can be recycled back to the cell surface. On the other hand, the cells were incubated with a noncovalent conjugate of AFP and 3H-arachidonic acid [3H-(20:4)], and the uptake of the fatty acid molecules studied by ultrastructural autoradiography. The cytoplasmic labeling, very low after an incubation in the presence of [3H-(20:4)]-AFP for 2 hours at 4 degrees C, increased rapidly after transfer of the cells for 5 minutes to 37 degrees C. These observations support the hypothesis that AFP plays a role in the intracellular delivery of polyunsaturated fatty acids.     

稀土元素在小麦体内分配行为的研究

采用水培,土培试验及中子活化分析技术,在作物生长效应曲线研究的基础上,系统地研究了稀土远征顷作物体内的含量、吸收、分布和转移等行为。所获结果表明,名稀土元素在作物体现人的分配行为受生物的内外因素与稀土来源、自身特征和元素间关系的影响,是作物稀土元素分配行为已有研究成果的重要补充与深化,并为土壤施用稀土元素提供促进一步的科学依据。     

Bioaccumulation of rare earth elements in cultured hela cells

HeLa S-3 cells were grown in minimal essential medium supplemented with 10% calf serum and 1 mM L-glutamine without adding any rare earth elements (REEs). Exponentially growing cells were collected, and dried materials were used to analyze their REE content by inductively coupled plasma-mass spectrometry. The results showed that the cells accumulated REEs in individually different manners; namely the accumulation ratio was higher in the lighter REEs than in the heavier REEs. To deduce the implication of the accumulation of REEs in HeLa cells, the accumulation ratios for REEs were compared with those of other biologically important elements. It was seen that the accumulation ratios obtained for REEs (from 31.8 [Ce] to 14.7 [Lu]) were intermediate among those of many bioelements: Fe (124), Mg (54.5), K (38.8), Cr (12.7), Na (11.8), Mn (11.3), Zn (10.7), Ca (8.8), and V (6.7).     

Rare earth elements in soil and in soil-grown plants

Concentrations of the rare earth elements (REEs) La, Ce, Nd, Sm, Eu, Gd, Tb, Yb and Lu were determined in leaves of 6 plant species (Norway spruce, silver fir, maple, ivy, blackberry, and wood fern), and in pertinent soils and soil extracts, also taken from the same site. The distribution of the individual REEs in plants showed little or no agreement with that in the soil or the soil extracts. Ce had a negative anomaly with respect to the soil in all plants. The REE distribution patterns of fir and spruce were almost identical, but differed profoundly from that of the other species. In most cases, concentration ratios between species were a smooth function of the atomic number of the REE. Very similar results were obtained at 2 additional sites.     

One of the possible mechanisms for the inhibition effect of Tb(III) on peroxidase activity in horseradish (<Emphasis Type="Italic">Armoracia rusticana</Emphasis>) treated with Tb(III)

One of the possible mechanisms for the inhibition effect of Tb(III) on peroxidase activity in horseradish (Armoracia rusticana) treated with Tb(III) was investigated using some biophysical and biochemical methods. Firstly, it was found that a large amount of Tb(III) can be distributed on the cell wall, that some Tb(III) can enter into the horseradish cell, indicating that peroxidase was mainly distributed on cell wall, and thus that Tb(III) would interact with horseradish peroxidase (HRP) in the plant. In addition, peroxidase bioactivity was decreased in the presence of Tb(III). Secondly, a new peroxidase-containing Tb(III) complex (Tb–HRP) was obtained from horseradish after treatment with Tb(III); the molecular mass of Tb–HRP is near 44 kDa and the pI is about 8.80. Thirdly, the electrocatalytic activity of Tb–HRP is much lower than that of HRP obtained from horseradish without treatment with Tb(III). The decrease in the activity of Tb–HRP is due to the destruction (unfolding) of the conformation in Tb–HRP. The planarity of the heme active center in the Tb–HRP molecule was increased and the extent of exposure of Fe(III) in heme was decreased, leading to inhibition of the electron transfer. The microstructure change in Tb–HRP might be the result of the inhibition effect of Tb(III) on peroxidase activity in horseradish.     

Measurement of markers for breast cancer in a model system using laser scanning cytometry

BACKGROUND: A variety of markers, including Ki67, estrogen receptors (ER), and progesterone receptors (PgR), are frequently measured in fine needle aspirates (FNA) from human breast carcinomas. We used a human breast carcinoma cell line, MCF7, as a model system to investigate the use of laser scanning cytometry (LSC) for the measurement of these markers. Additionally, we measured the number of apoptotic cells. METHODS: Cells were treated with drugs to vary the expression of markers and the number of apoptotic cells. They were then fixed on microscope slides. For LSC, the cells were stained for the different markers with fluorescein using immunofluorescence and for apoptotic cells using the TUNEL assay. The nuclei were counterstained with propidium iodide. A parallel set of slides was stained using horseradish peroxidase and diaminobenzidine and scored manually by conventional light microscopy. RESULTS: The results from the LSC closely paralleled those obtained by manual scoring of immunohistochemical stains. CONCLUSIONS: It should be possible to use LSC for the routine measurement of nuclear markers in FNAs from human breast carcinomas.     

甜椒始花期吸收的氮素在体内的动态分配规律

 通过水培试验,利用15N示踪技术研究了甜椒(Capsicum annuum L.)始花期吸收的氮素在体内的动态分配规律。结果表明：甜椒营养器官干物质积累动态呈快速直线增长趋势,而果实则从盛采前期开始 超过其它器官。根与果实含氮量从始采期开始一直维持在同一水平;叶片中氮的含量则随生长发育而不断下降。在果实盛采期,叶片、果实和根中的氮素含量差异不明显,但是均比茎和侧枝高两倍。甜椒植株各器官的氮素积累动态与干物质积累动态相似。在始花期通过根吸收的大部分标记态氮首先分配、贮存在叶片中,吸收后第二周发现有１/2的标记态氮运转到了植物的新生部分,至吸收后的第四周标记态氮运转显著下降,但是,在吸收后的第六周仍然发现有小部分标记态氮运转到了新生部分。说明新近吸收的氮素在甜椒体内的再运转相对容易,其再利用率也高,随着在植物体内某器官存在时间的延长其再利用率逐渐降低。在整个果实收获期间,甜椒不同器官之间以果实对氮素的竞争力最强。在整株水平上,始花期吸收的标记态氮从营养器官向生殖器官的运转,既受到氮素竞争库（果实）强度和容量的影响,也受到时间因素的制约。     

Translocation of Bax to mitochondria during apoptosis measured by laser scanning cytometry

BACKGROUND: During induction of apoptosis, the pro-apoptotic member of the Bcl-2 protein family (Bax) undergoes translocation to the mitochondria. The translocation, which leads to accumulation of Bax in the mitochondrial intermembrane space, appears to be the critical event determining release of cytochrome c to cytosol: the latter event triggers the irreversible steps of apoptosis, namely, the activation of caspases and the initiation of the degradation of many proteins. The aim of this study was to utilize the morphometric capabilities of the laser scanning cytometer (LSC) and adapt this instrument to detect and measure in situ the process of translocation of Bax to mitochondria. METHODS: Human breast carcinoma MCF-7 cells growing on microscope slides were treated with the DNA topoisomerase I inhibitor, camptothecin (CPT). CPT is known to induce apoptosis preferentially of S-phase cells. The cells were fixed and permeabilized on the slides, their DNA was stained with propidium iodide (PI), Bax was detected immunocytochemically with the fluoresceinated antibody, and red and green fluorescence emission was measured by the LSC. RESULTS: Prior to induction of apoptosis, Bax was uniformly and diffusely dispersed in the cell nucleus and cytoplasm. Its translocation and accumulation in mitochondria in cells undergoing apoptosis were detected and measured by the LSC as the increase in intensity of maximal pixel of Bax immunofluorescence. Bivariate analysis of DNA content versus maximal pixel of Bax fluorescence revealed that the CPT-induced Bax translocation into mitochondria was preferential to S-phase cells. Total cellular Bax immunofluorescence measured by flow cytometry, however, was increased in all phases of the cycle without a preference to S-phase cells. CONCLUSION: Changes in abundance and localization of particular proteins that undergo translocation within the cell, leading to their altered local density, may be conveniently detected by the LSC by taking advantage of its morphometric capabilities. Measurement of total cellular Bax immunofluorescence by flow cytometry combined with analysis of its translocation by LSC revealed that apoptosis of S-phase cells induced by CPT was unrelated to overall Bax abundance per cell but correlated with its accumulation in mitochondria.     

放射性铈在模拟水生生态系中的行为

研究＾１４１Ｃｅ在水－底泥－水生动植物系统中的迁移和消长动态,结果表明,系统各组分中＾１４１Ｃｅ浓度与时间关系由多项指数描述,其中池水中的＾１４１Ｃｅ浓度随时间延长单调地降低,底泥中＾１４１Ｃｅ浓度随时间延长增加,水生动植物中＾１４１Ｃｅ浓度则各自经历某一最大值后逐渐降低,水生动植物对＾１４１Ｃｅ皆有一定的富集能力,其大小次序依次为：金鱼藻〉螺狮〉鱼,而鱼体中的＾１４１Ｃｅ主要集中内脏,螺壳与螺肉     

Non-invasive microelectrode cadmium flux measurements reveal the spatial characteristics and real-time kinetics of cadmium transport in hyperaccumulator and nonhyperaccumulator ecotypes of Sedum alfredii

This study aims to determine the spatial characteristics and real-time kinetics of cadmium transport in hyperaccumulator (HE) and non hyperaccumulator (NHE) ecotypes of Sedum alfredii using a non-invasive Cd-selective microelectrode. Compared with the NHE S. alfredii, the HE S. alfredii showed a higher Cd influx in the root apical region and root hair cells, as well as a significantly higher Cd efflux in the leaf petiole after root pre-treatment with cadmium chloride (CdCl₂). Thus, HE S. alfredii has a higher capability for the translocation of absorbed Cd to the shoot. Moreover, the mesophyll tissues, isolated mesophyll protoplasts, and intact vacuoles from HE S. alfredii exhibited an instantaneous influx of Cd in response to CdCl₂ treatment with mean rates that are markedly higher than those from NHE S. alfredii. Therefore, the hyper-accumulating trait of HE S. alfredii is characterized by the rapid Cd uptake in specific root regions, including the apical region and root hair cells, as well as by the rapid root-to-shoot translocation and the highly efficient Cd-permeable transport system in the plasma membrane and mesophyll cell tonoplast. We suggest that the non-invasive Cd-selective microelectrode is an excellent method with a high degree of spatial resolution for the study of Cd transport at the tissue, cellular, and sub-cellular levels in plants.     

Leaf structure and translocation in sugar beet

Anatomical and ultrastructural details of a translocating 10-cm leaf of sugar beet (Beta vulgaris L. var. Klein Wanzleben) were correlated with translocation rate data. The minor veins were found to be 13 times as extensive as the major veins and measure 70 cm/cm² leaf lamina. Measurements disclosed that a 33-μ length of minor vein services 29 mesophyll cells with the result that translocate moves an average of 73 μ or 2.2 cell diameters during transport from mesophyll cells to a minor vein. High-resolution, freeze-dry autoradiography revealed that assimilates accumulate in organelle-rich cells of the minor vein phloem. Correlation of phloem volume and loading rate for minor veins yielded an uptake rate of 735 μmoles of sucrose per g fresh weight of phloem. The arrangement and structural features of minor veins appeared to be consistent with the concept that vein loading precedes translocation.     

Bioaccessibility and health risk assessment of rare earth elements in Porphyra seaweed species

In this study, the content and bioaccessibility of rare earth elements (REEs) in Porphyra spp. Samples before and after thermal treatments were investigated. Additionally, the risks of REEs exposure to human health were assessed. REEs content significantly reduced after thermal processing (P < 0.01), and the removal rate of REEs was approximately 30%. Thermal treatment increased REEs bioaccessibility from 44% to 64.34%. The concentration and bioaccessibility of Ce, La, Y, Nd were high in raw and thermally treated Porphyra samples, and there was no correlation between REEs content and bioaccessibility. Based on the following parameters: highest content of REEs in the studied seaweed samples (13.45 mg/kg), the highest daily seafood consumption (44.9 g/day), and the highest bioaccessibility (64.34%), the ratio of the calculated daily intake (DI) to daily allowable intake by diet (DAI_diet) of REEs did not exceed the reference value in rare earth mining areas or under extreme conditions. The DI via seafood consumption would be exceeded when the content of REEs in the seafood sample is greater than 15.77 mg/kg. In this study, the concentration of REEs did not exceed 15.77 mg/kg in any sample. Thus, the human health risks of REEs associated with seafood are low.