Stacking Structure of Vaterite Revealed by Atomic Imaging and Diffraction Analysis.

Anhydrous calcium carbonate crystals exist as three polymorphs: calcite, aragonite, and vaterite. Although vaterite is a metastable phase rarely found in the geological environment, it is intriguing that various biominerals are composed of vaterite. The processes of stable vaterite formation in biological systems cannot be understood without elucidating the nature of vaterite. The crystal structure of vaterite has been discussed for nearly a century but is still an open question. Here we propose the actual structure of vaterite by combining atomic imaging and diffraction analysis with simulations of disordered stacking sequences. Vaterite basically appears as layers of hexagonal calcium planes and carbonate (CO32-)-containing sheets stacked with +60°, -60°, or 180° rotations from the underlying layer. However, equivalent carbonate positions in alternating layers are forbidden, and four-layer stacking in which the fourth layer rotates 180° relative to the first layer are predominant, forming an orthogonal reciprocal lattice in diffraction patterns. These stacking characteristics replicate the intensity distribution in the electron and X-ray diffraction patterns. This study has almost completely elucidated the crystal structure and stacking sequence of vaterite. Our findings provide insights into the thermodynamic stability of vaterite, which facilitates comprehension of the biomineralization processes and growth dynamics of calcium carbonate.

Effect of phosphoproteins on intracellular calcification of bacteria.

This study aimed to evaluate the effects of phosphoproteins on bacterial mineralization. Dental calculus formation is attributed to bacterial mineralization in the oral cavity; however, the influence of phosphoproteins (which are abundant in saliva) is not clear. The model bacterium Escherichia coli was suspended in a calcification solution containing casein as a model phosphoprotein. To evaluate mineralization independent of bacterial metabolism, bacteria killed by heat treatment at 70°C were compared with viable bacteria. After incubation at 37°C for 24 h, the mode of calcification was observed using electron microscopy and energy dispersive x-ray spectroscopy. Solutions without casein produced precipitation in solution, which was identical to that in experiments without bacteria. In contrast, calcification solutions with 200 ppm casein only produced calcium phosphate deposition intracellularly. Without heat treatment, intracellular calcification rarely occurred, even when casein was added. Thus, phosphoproteins promoted intracellular calcification of dead bacteria; this is similar to the calcification of insoluble matrices, such as collagen fibrils, promoted by acidic polymers. We concluded that intracellular calcification is caused by the collagen fibril-like behavior of dead bacteria. The promotion of intracellular calcification of dead bacteria by phosphoproteins suggested a basic principle of dental calculus formation.

Dissolution behavior of radiocesium-bearing microparticles as a function of solution compositions.

More than a decade has passed since the Fukushima nuclear accident in 2011 and contamination around the nuclear power plant is primarily caused by 137Cs. One of the materials retaining radiocesium in the environment is radiocesium-bearing silicate glass microparticles (CsMPs), which have not been reported in previous nuclear accidents. Although the prediction of environmental fates of CsMPs is of interest because of their extremely high specific radioactivity, knowledge about their physicochemical properties is still limited. Here we show that the dissolution behavior of CsMPs is comparable to that of silica-rich glass and significantly depends on the surrounding environment. CsMP dissolution experiments were conducted in solutions with various solute components and pH levels at 60 °C. In neutral and basic solutions, the estimated dissolution rate was accelerated by alkali ions such as Na+, which is known to play a catalytic role for the dissolution of silica. In contrast, the dissolution in acid was slow even in the presence of alkali ions. The dissolution under acid conditions was possibly retarded by a thin amorphous silica layer formed on the CsMP surfaces. Such characteristics of the dissolution are consistent with that of silica-rich glass. To infer the dissolution behavior of CsMPs in the human body, the dissolution rate in Ringer's solution at 37 °C was estimated as 1.00 ± 0.37 µm/year.

A simple multilevel sampler for synchronous collection of stratified waters.

Stratified water collection plays a crucial role in water quality monitoring, as most water bodies are not perfectly mixed systems. In order to precisely collect stratified waters, we developed an inexpensive, simple, and high-resolution sampler to simultaneously collect and measure physical and chemical parameters along vertical water profiles. The water sampler predominantly consists of two parts: (1) an apparatus for measuring sampling depth below the water and (2) water sampling units secured below the water. Proof of concept water sampling was performed in Caohai wetland (Southwest China) at 40 cm intervals, as sampling depth and interval are adjustable. Stratified waters in four sampling sites were characterized by markedly different levels of major and trace elements as well as physicochemical parameters. Results indicate this simple multilevel sampler to be a cheap, precise, and portable option for simultaneously collecting water samples at different depths in a wide array of water body types.

Deposition of Lead Phosphate by Lead-Tolerant Bacteria Isolated from Fresh Water near an Abandoned Mine.

Specialist bacteria can synthesize nanoparticles from various metal ions in solution. Metal recovery with high efficiency can be achieved by metal-tolerant microorganisms that proliferate in a concentrated metal solution. In this study, we isolated bacteria (Pseudomonas sp. strain KKY-29) from a bacterial library collected from water near an abandoned mine in Komatsu City, Ishikawa Prefecture, Japan. KKY-29 was maintained in nutrient medium with lead acetate and synthesized hydrocerussite and pyromorphite nanoparticles inside the cell; KKY-29 also survived nanoparticle synthesis. Quantitative PCR analysis of genes related to phosphate metabolism showed that KKY-29 decomposed organic phosphorus to synthesize lead phosphate. KKY-29 also deposited various metal ions and synthesized metal nanoparticles when incubated in various metal salt solutions other than lead. The present study considers the development of biotechnology to recover lead as an economically valuable material.

Functional analyses of chitinolytic enzymes in the formation of calcite prisms in Pinctada fucata.

The mollusk shells present distinctive microstructures that are formed by small amounts of organic matrices controlling the crystal growth of calcium carbonate. The shell of Pinctada fucata has the prismatic layer consisting of prisms of single calcite crystals and the nacreous layer consisting of aragonite tablets. The calcite crystal of prisms contains small angle grain boundaries caused by a dense intracrystalline organic matrix network to improve mechanical strength. Previously, we identified chitin and chitinolytic enzymes as components of this intracrystalline organic matrix. In this study, to reveal the function of those organic matrices in calcium carbonate crystallization, calcites synthesized in chitin gel with or without chitinolytic enzymes were analyzed by using transmission electron microscope (TEM) and atom probe tomography (APT), showing ion clusters derived from chitin inside of a calcite and small angle grain boundaries at optimal chitinolytic concentration. Furthermore, we performed the experiment in which chitinase inhibitor was injected into a living P. fucata. Nanoindentation and electron back scattered diffraction (EBSD) show that mechanical properties and crystal orientation were changed. These results suggested that chitinolytic enzymes work cooperatively with chitin to regulate the crystal growth and mechanical properties of the prismatic layer.

Microstructure and mineral components of the outer dentin of Chimaera phantasma tooth plates.

Tooth plates are a unique dental organ found in holocephalan fishes and lungfish. The chimaeroid tooth plates are atypical in terms of biomineralization, due to the hard tissue composition of whitlockite and apatite, while those of lungfish and other vertebrates are composed of apatite. The tooth plates are overlaid by a thin veneer-outer dentin-whose composition and role are not known. We aimed to test whether the outer dentin is composed of whitlockite or apatite, and whether it protects the osteodentin from abrasion and supports its overall strength. For this purpose, the mineral components and microstructure of outer dentin were studied. Our analyses of the outer dentin from the anterior (vomerine) tooth plates of Chimaera phantasma revealed that the mineral component is magnesium- and carbonate-containing calcium-deficient apatite and that the outer dentin has a three-zone structure. The main body is sandwiched between thin zones, which are less mineralized than the main body. Furthermore, in the outer zone and the main body, a higher-order structure was formed in accordance with the organization of wide and narrow fibers. Mineralization made the main body a composite of bundles of fibers and apatite. Transmission electron microscopy showed a structural relationship between apatite and the fibrous component on which the apatite was formed. Such a structure of the main body could be highly effective as a framework to resist abrasion and support the overall strength of the tooth plate.

Identification and quantification of contributions to karst groundwater using a triple stable isotope labeling and mass balance model.

Although karst groundwater systems provide critical ecosystem services in many regions worldwide, anthropogenic contamination has seriously degraded groundwater quality. Properly elucidating geochemical processes, quantifying contributions of natural and anthropogenic end members, and then protecting karst aquifer systems remain challenging from scientific and engineering aspects. To identify the hydrochemical processes and quantifying contributions of end members (especially, contamination end members), 49 samples were collected from cave waters (CW), artesian springs (AS), and gravity springs (GS) in a karst watershed in Guiyang, China. With increased anthropogenic contamination, the CW, AS, and GS characterized by a Ca-Mg-SO42--HCO3- composition often had pH and SO42- concentrations exceeding USEPA secondary drinking water standards. That is attributed to the influence of water-rock interaction, rainfall, and anthropogenic sources (mainly, sewage and fertilizers), in agreement with the compositions of δ34SSO4, δ18OSO4, and 87Sr/86Sr as well as the results of principal component analysis and correlation coefficients. Based on an end-member mixing model, contributions of rainfall and anthropogenic sources were 47% and 33% of GS, 52% and 41% of CW, and 58% and 35% of AS, respectively. It suggests that the karst groundwater quality is predominantly controlled by rainfall and anthropogenic sources (especially, land use). Results may be applied to properly evaluate the impacts of natural and anthropogenic sources on karst aquifers, coupled with actions to efficiently control potential contamination end members.

Author Correction: Formation of gold nanoparticles by glycolipids of Lactobacillus casei.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Reactor environment during the Fukushima nuclear accident inferred from radiocaesium-bearing microparticles.

Radiocaesium-bearing microparticles (CsMPs), which are substantially silicate glass, were formed inside the damaged reactor and released to the environment by the Fukushima Dai-ichi Nuclear Power Plant accident in March 2011. The present study reports several valuable findings regarding their composition and structure using advanced microanalytical techniques. X-ray absorption near-edge structure of Fe L3-absorption indicated that the oxidation state of the iron dissolved in the glass matrix of the CsMPs was originally nearly divalent, suggesting that the atmosphere in which the CsMPs were formed during the accident was considerably reductive. Another major finding is that sodium, which has not been recognised as a constituent element of CsMPs thus far, is among the major elements in the glass matrix. The atomic percent of Na is higher than that of other alkali elements such as K and Cs. Furthermore, halite (NaCl) was found as an inclusion inside a CsMP. The existence of Na in CsMPs infers that seawater injected for cooling might reach the inside of the reactor before or during the formation of the CsMPs. These results are valuable to infer the environment inside the reactor during the accident and the debris materials to be removed during the decommissioning processes.

Tracing and quantifying contributions of end members to karst water at a coalfield in southwest China.

Karst water, which provides 25% of the world's drinking water, is especially vulnerable to anthropogenic contamination. Such is the case in southwestern China with trace element pollution in important karst aquifers. Approximately 20% of the total study area consisted of abandoned mine tailings with elevated concentrations of Fe, S, Mn, As, Cu, and Cr. Acid mine drainage (AMD) water originating from pyrite oxidation of the tailings was characterized by low pH and high concentrations of Fe, SO42-, and As. Concentrations of Fe, Mn, Al, SO42-, As, Cd, and Pb in spring water in wet and dry seasons were greater than WHO and USEPA drinking water guidelines. Based on the results of mineral characterizations, hydrogeochemistry, and isotopic compositions (δ34SSO4, δ18OSO4, and δ18OH2O), the chemistry of AMD water was primarily controlled by pyrite oxidation, river water by atmospheric precipitation, and spring water by carbonate rock dissolution and mixing with river and AMD waters. A three-end-member mixing model identified the contributions of these different end members to spring water quality. Although AMD water was characterized by the lowest mixing percentages during the wet (14.1%) and dry (26.9%) seasons, it played a very important role in degrading spring water quality. Based on these findings, an investigation strategy was developed for illuminating seasonal water quality and potential remediation methods corresponding to the contaminants in the spring water are also proposed to manage this seriously polluted karst system. Results could benefit remediation planning for these distinctively complex and vulnerable systems in other regions of the world.

A review of Cs-bearing microparticles in the environment emitted by the Fukushima Dai-ichi Nuclear Power Plant accident.

Scientists face challenge in identifying the radioactive materials which are found as dotted images on various imaging plate (IP) autoradiographic photos of radioactively contaminated materials by the Fukushima Dai-ichi Nuclear Power Plant (F1NPP, or FDNPP) accident, such as air filter, fugitive dust, surface soil, agricultural materials, and water-shed samples. It has been revealed that they are minute particles with distinct morphology and elemental composition with high specific radioactivity, and different from those of the so-called Chernobyl hot particles. Basically, they are glassy particles once molten, composed of Si, O, Fe, Zn etc. with highly concentrated radiocaesium, which can be called as radiocaesium-bearing microparticles (CsMP). At present, CsMP can be classified into two types, Types-A and -B, which are characterized by different specific radioactivity, 134Cs/137Cs ratio, size and morphology, and geographic distribution around F1NPP. Such studies on the CsMP from various aspects have provided valuable information about what happened in the nuclear reactors during the F1NPP accident and fates of the CsMP in the environment. This review first provides a retrospective view on the research history of the CsMP, which is helpful to understand the unique character of the CsMP. Subsequently, more details about the current understanding of the natures of these hot particles, such as origin, morphology, chemical compositions, thermal properties, water-solubility, and secondary migration of CsMP in river and ocean systems are described with future prospects.

Dissolution behaviour of radiocaesium-bearing microparticles released from the Fukushima nuclear plant.

Radiocaesium-bearing microparticles (CsMPs) composed of silicate glass were released by the Fukushima Daiichi Nuclear Power Plant accident in March 2011. Since CsMPs contain a high concentration of radiocaesium, their dynamics and fate in the environment are urgent issues to be investigated. Here, we show that CsMPs are dissolved by weathering in the environment and that their radioactivity is more rapidly decreased by dissolution than the physical decay of radiocaesium. We conducted dissolution experiments with CsMPs in pure water that absorbed CO2 from the atmosphere and in artificial seawater at several temperatures. The dissolution progress was monitored by the decrease in the 137Cs radioactivity in CsMPs, and the dissolution rate was estimated. The activation energy for the dissolution of CsMPs was estimated to be 65 and 88 kJ/mol and the dissolution rate at 13 °C (approximate annual mean temperature in Fukushima City) was 0.014 and 0.140 µm/y for pure water and seawater, respectively, assuming that radiocaesium is uniformly distributed in spherical CsMPs. The shapes of the CsMPs dissolved in pure water were considerably altered, suggesting that the dissolution proceeded without maintaining the geometry. Tin oxide and iron oxide nanoparticulates formed on the surfaces. Such features were similar to those observed in CsMPs recently collected in Fukushima Prefecture, indicating that weathering dissolution of CsMPs is also occurring in the environment. For the CsMPs dissolved in seawater, a crust of secondary Mg- and Fe-rich minerals was formed, and the glass matrix inside the crust decreased, creating space between the crust and the glass matrix.

Inner structure and inclusions in radiocesium-bearing microparticles emitted in the Fukushima Daiichi Nuclear Power Plant accident.

Radiocesium-bearing microparticles (CsMPs), consisting substantially of silicate glass, were released to the environment during the Fukushima Daiichi Nuclear Power Plant accident in March 2011. Since the CsMPs were formed inside the damaged reactors during the accident, we investigate the inner structures of several CsMPs by transmission electron microscopy to understand the events within the reactors. Elemental mapping of the CsMPs shows a distinct radial distribution of Cs with a higher concentration near the surface of the CsMPs, implying that Cs was in a gaseous state in the reactor atmosphere and diffused into the glass matrix after formation of the glass particles. In some CsMPs, Zn and Fe also showed a similar radial distribution to Cs, suggesting that those elements may have diffused outward where Cs was abundant. In addition, submicron crystals were present as inclusions in several of the CsMPs and were identified as chromium spinels ((Fe2+,Zn)(Cr,Fe3+)2O4), acanthite (Ag2S), molybdenite (MoS2) and hessite (Ag2Te). The spinels contained ferrous iron (Fe2+), suggesting that the atmosphere inside the reactors was reductive to some extent. Also, boron was not detected in the glass matrix of the CsMPs despite using electron energy-loss spectroscopy, indicating that most of the control rods made of B4C might have created a eutectic alloy without vaporization. These detailed investigations of the inner structures in the CsMPs may offer information on the damaged reactors that are difficult to access because of the high radiation fields.

Loss of radioactivity in radiocesium-bearing microparticles emitted from the Fukushima Dai-ichi nuclear power plant by heating.

Radiocesium-bearing microparticles (CsPs) substantially made of silicate glass are a novel form of radiocesium emitted from the broken containment vessel of Fukushima Dai-ichi nuclear power plant. CsPs have a potential risk of internal radiation exposure caused by inhalation. Radiation-contaminated waste including CsPs is being burned in incinerators; therefore, this study has investigated the responses of CsPs to heating in air. The radioactivity of CsPs gradually decreased from 600 °C and was almost lost when the temperature reached 1000 °C. The size and spherical morphology of CsPs were almost unchanged after heating, but cesium including radiocesium, potassium and chlorine were lost, probably diffused away from the CsPs. Iron, zinc and tin originally dissolved in the glass matrix were crystallized to oxide nanoparticles inside the CsPs. When the CsPs were heated together with weathered granitic soil that is common in Fukushima, the radiocesium released from CsPs was sorbed by the surrounding soil. From these results, it is expected that the radioactivity of CsPs will be lost when radiation-contaminated waste including CsPs is burned in incinerators.

Cesium desorption behavior of weathered biotite in Fukushima considering the actual radioactive contamination level of soils.

For the better understanding of radioactive contamination in Fukushima Prefecture at present and in future, Cs desorption experiments have been conducted mainly using weathered biotite (WB) collected from Fukushima Prefecture and considering the actual contamination level (∼10-10 wt%) of radiocesium in Fukushima Prefecture. In the experiments, 137Cs sorbed to WB by immersing in 137Cs solution for one day was mostly desorbed by solutions of 1 M NaNO3, 1 M LiNO3, 10-1 M HCl, and 10-1 M HNO3, although it was barely desorbed by 1 M KNO3, 1 M CsNO3, 1 M NH4NO3, and natural seawater. X-ray diffraction analysis of WB after immersing in these solutions suggested that the collapse of the hydrated interlayers in WB suppressed the desorption of Cs. On the other hand, 137Cs was barely desorbed from WB even by the treatments with solutions of NaNO3 and LiNO3 if the duration for the sorption was longer than approximately two weeks, as well as radioactive WB collected from actual contaminated soils in Fukushima Prefecture. This result implies that Cs sorbed in WB became more strongly fixed with time. Probably removal of radiocesium sorbed in weathered granitic soil at Fukushima Prefecture is difficult by any electrolyte solutions, as more than seven years have passed since the accident.

Involvement of Acidic Polysaccharide Ph-PS-2 and Protein in Initiation of Coccolith Mineralization, as Demonstrated by In Vitro Calcification on the Base Plate.

Coccolithophorids, unicellular marine microalgae, have calcified scales with elaborate structures, called coccoliths, on the cell surface. Coccoliths generally comprise a base plate, CaCO3, and a crystal coat consisting of acidic polysaccharides. In this study, the in vitro calcification conditions on the base plate of Pleurochrysis haptonemofera were examined to determine the functions of the base plate and acidic polysaccharides (Ph-PS-1, -2, and -3). When EDTA-treated coccoliths (acidic polysaccharide-free base plates) or low pH-treated coccoliths (whole acidic polysaccharide-containing base plates) were used, mineralization was not detected on the base plate. In contrast, in the case of coccoliths which were decalcified by lowering of the pH and then treated with urea (Ph-PS-2-containing base plates), distinct aggregates, probably containing CaCO3, were observed only on the rim of the base plates. Energy dispersive X-ray spectroscopy (EDS) confirmed that the aggregates contained Ca and O, although X-ray diffraction analysis did not reveal any evidence of crystalline materials. Also, in vitro mineralization experiments performed on EDTA-treated coccoliths using isolated acidic polysaccharides demonstrated that the Ca-containing aggregates were markedly formed only in the presence of Ph-PS-2. Furthermore, in vitro mineralization experiments conducted on protein-extracted base plates suggested that the coccolith-associated protein(s) are involved in the Ca deposition. These findings suggest that Ph-PS-2 associated with the protein(s) on the base plate rim initiates Ca2+ binding at the beginning of coccolith formation, and some other factors are required for subsequent calcite formation.

Structural and functional analyses of a TIMP and MMP in the ligament of Pinctada fucata.

The bivalve hinge ligament is the hard tissue that functions to open and close shells. The ligament contains fibrous structures consisting of aragonite crystals surrounded by a dense organic matrix. This organic matrix may contribute to the formation of fibrous aragonite crystals, but the mechanism underlying this formation remains unclear. In this study, we identified a novel ligament-specific protein, Pinctada fucata tissue inhibitor of metalloproteinase (PfTIMP), from the fibrous organic matrix between aragonite crystals in the ligament using the amino acid sequence and cDNA cloning methods. PfTIMP consists of 143 amino acid residues and has a molecular weight of 13,580.4. To investigate the activity of PfTIMP, inhibition of matrix metalloproteinase (MMP) activity was measured. PfTIMP strongly inhibited human MMP13 and MMP9. Eight MMP homologs were identified from a P. fucata genomic database by BLAST search. To identify the specific MMP that may contribute to ligament formation, the expression level of each MMP was measured in the mantle isthmus, which secretes the ligament. The expression of MMP54089 increased after scratching of the ligament, while the expressions of other MMPs did not increase after doing the same operation. To identify the role of MMP54089 in forming the ligament structure, double stranded (ds) RNA targeting MMP54089 was injected into living P. fucata to suppress the function of MMP54089. Scanning electron microscopic images showed disordered growing surfaces of the ligament in individuals injected with MMP54089-specific dsRNA. These results suggest that PfTIMP and MMP54089 play important roles in the formation of the fibrous ligament structure.

Crystal defects induced by chitin and chitinolytic enzymes in the prismatic layer of Pinctada fucata.

Biomineralization, in which organisms create biogenic hard tissues, with hardness or flexibility enhanced by organic-inorganic interaction is an interesting and attractive focus for application of biomimetic functional materials. Calcites in the prismatic layer of Pinctada fucata are tougher than abiotic calcites due to small crystal defects. However, the molecular mechanism of the defect formation remains unclear. Here, chitin and two chitinolytic enzymes, chitinase and chitobiase, were identified as organic matrices related to for the formation of small crystal defects in the prismatic layer. Experiments with a chitinase inhibitor in vivo showed chitinase is necessary to form the prismatic layer. Analysis of calcite crystals, which were synthesized in a chitin hydrogel treated with chitinolytic enzymes, by electron microscopy and X-ray diffraction showed that crystal defects became larger as chitin was more degraded. These results suggest that interactions between chitin and calcium carbonate increase as chitin is thinner.

Weak size dependence of resuspended radiocesium adsorbed on soil particles collected after the Fukushima nuclear accident.

Most studies of the properties of airborne radionuclides emitted from the Fukushima Daiichi Nuclear Power Plant have focused on the relatively early stages of the accident, and little is known about the characteristics of radiocesium in the long-term. In this study, we analyzed activity size distributions of airborne radiocesium collected over 5 months in Tsukuba, Japan. Radiocesium in the accumulation mode size range (0.1-2 µm in aerodynamic diameter) was overwhelming in the early aerosol samples and decreased with time, while that associated with coarse aerosols remained airborne. We examined the radiocesium adsorbed onto airborne soil particles, and found that the size dependence of 137Cs surface density adsorbed on soil particles was weak. That is, radiocesium was distributed homogeneously throughout the aerodynamic diameter range of 2.1-11 µm. This characteristic may be related to the reported structure of radiocesium-bearing soil particles collected from the ground, which consisted of an aggregate of specific clay minerals and other non-cesium adsorbing particles. The resuspension factors for the first two aerosol samples collected during late April and May 2011 were close to those in European cities in the months following the Chernobyl accident, despite different soil and weather conditions.