Yasuto Matsui

Finite element 3D reconstruction of the pulmonary acinus imaged by synchrotron X-ray tomography

The alveolated structure of the pulmonary acinus plays a vital role in gas exchange function. Three-dimensional (3D) analysis of the parenchymal region is fundamental to understanding this structure-function relationship, but only a limited number of attempts have been conducted in the past because of technical limitations. In this study, we developed a new image processing methodology based on finite element (FE) analysis for accurate 3D structural reconstruction of the gas exchange regions of the lung. Stereologically well characterized rat lung samples (Pediatr Res 53: 72-80, 2003) were imaged using high-resolution synchrotron radiation-based X-ray tomographic microscopy. A stack of 1,024 images (each slice: 1024 x 1024 pixels) with resolution of 1.4 mum(3) per voxel were generated. For the development of FE algorithm, regions of interest (ROI), containing approximately 7.5 million voxels, were further extracted as a working subunit. 3D FEs were created overlaying the voxel map using a grid-based hexahedral algorithm. A proper threshold value for appropriate segmentation was iteratively determined to match the calculated volume density of tissue to the stereologically determined value (Pediatr Res 53: 72-80, 2003). The resulting 3D FEs are ready to be used for 3D structural analysis as well as for subsequent FE computational analyses like fluid dynamics and skeletonization.

Fecal Source Tracking in Water by Next-Generation Sequencing Technologies Using Host-Specific Escherichia coli Genetic Markers

High levels of fecal bacteria are a concern for the aquatic environment, and identifying sources of those bacteria is important for mitigating fecal pollution and preventing waterborne disease. Escherichia coli has been used as an indicator of fecal pollution, however less success has been achieved using this organism for library-independent microbial source tracking. In this study, using next-generation sequencing technology we sequenced the whole genomes of 22 E. coli isolates from known sources (9 from humans, 2 from cows, 6 from pigs, and 5 from chickens) and identified candidate host-specific genomic regions. Specificity testing on the candidate regions was performed using 30 E. coli isolates from each source. Finally, we identified 4 human-, 2 cow-, 3 pig-, and 4 chicken-specific genetic markers useful for source tracking. We also found that a combination of multiplex PCR and dual index sequencing is effective for detecting multiple genetic markers in multiple isolates at one time. This technique was applied to investigating identified genetic markers in 549 E. coli isolates obtained from the Yamato River, Japan. Results indicate that humans constitute a major source of water contamination in the river. However, further work must include isolates obtained from geographically diverse animal hosts to make this method more reliable.

Future projection of radiocesium flux to the ocean from the largest river impacted by Fukushima Daiichi Nuclear Power Plant

Following the initial fall out from Fukushima Dai-ichi Nuclear Power Plant (FDNPP), a significant amount of radiocesium has been discharged from Abukuma River into the Pacific Ocean. This study attempted to numerically simulate the flux of radiocesium into Abukuma River by developing the multiple compartment model which incorporate the transport process of the radionuclide from the ground surface of the catchment area into the river, a process called wash off. The results from the model show that the sub-basins with a high percentage of forest area release the radionuclides at lower rate compared to the other sub-basins. In addition the results show that the model could predict the seasonal pattern of the observed data. Despite the overestimation observed between the modeled data and the observed data, the values of R2 obtained from 137Cs and 134Cs of 0.98 and 0.97 respectively demonstrate the accuracy of the model. Prediction of the discharge from the basin area for 100 years after the accident shows that, the flux of radiocesium into the Pacific Ocean is still relatively high with an order of magnitude of 109 bq.month−1 while the total accumulation of the discharge is 111 TBq for 137Cs and 44 TBq for 134Cs.

Characterization of Pathogenic Escherichia coli in River Water by Simultaneous Detection and Sequencing of 14 Virulence Genes

The occurrence of pathogenic Escherichia coli in environmental waters increases the risk of waterborne disease. In this study, 14 virulence genes in 669 E. coli isolates (549 isolates from the Yamato River in Japan, and 30 isolates from each of the following hosts: humans, cows, pigs, and chickens) were simultaneously quantified by multiplex PCR and dual index sequencing to determine the prevalence of potentially pathogenic E. coli. Among the 549 environmental isolates, 64 (12%) were classified as extraintestinal pathogenic E. coli (ExPEC) while eight (1.5%) were classified as intestinal pathogenic E. coli (InPEC). Only ExPEC-associated genes were detected in human isolates and pig isolates, and 11 (37%) and five (17%) isolates were classified as ExPEC, respectively. A high proportion (63%) of cow isolates possessed Shiga-toxin genes (stx1 or stx2) and they were classified as Shiga toxin-producing E. coli (STEC) or enterohemorrhagic E. coli (EHEC). Among the chicken isolates, 14 (47%) possessed iutA, which is an ExPEC-associated gene. This method can determine the sequences as well as the presence/absence of virulence genes. By comparing the sequences of virulence genes, we determined that sequences of iutA were different among sources and may be useful for discriminating isolates, although further studies including larger numbers of isolates are needed. Results indicate that humans are a likely source of ExPEC strains in the river.

Characterization and source profiling of volatile organic compounds in indoor air of private residences in Selangor State, Malaysia

Volatile Organic Compounds (VOCs) in indoor air were investigated at 39 private residences in Selangor State, Malaysia to characterize the indoor air quality and to identify pollution sources. Twenty-two VOCs including isomers (14 aldehydes, 5 aromatic hydrocarbons, acetone, trichloroethylene and tetrachloroethylene) were collected by 2 passive samplers for 24h and quantitated using high performance liquid chromatography and gas chromatography mass spectrometry. Source profiling based on benzene/toluene ratio as well as statistical analysis (cluster analysis, bivariate correlation analysis and principal component analysis) was performed to identify pollution sources of the detected VOCs. The VOCs concentrations were compared with regulatory limits of air quality guidelines in WHO/EU, the US, Canada and Japan to clarify the potential health risks to the residents. The 39 residences were classified into 2 groups and 2 ungrouped residences based on the dendrogram in the cluster analysis. Group 1 (n=30) had mainly toluene (6.87±2.19μg/m3), formaldehyde (16.0±10.1μg/m3), acetaldehyde (5.35±4.57μg/m3) and acetone (11.1±5.95μg/m3) at background levels. Group 2 (n=7) had significantly high values of formaldehyde (99.3±10.7μg/m3) and acetone (35.8±12.6μg/m3), and a tendency to have higher values of acetaldehyde (23.7±13.5μg/m3), butyraldehyde (3.35±0.41μg/m3) and isovaleraldehyde (2.30±0.39μg/m3). The 2 ungrouped residences showed particularly high concentrations of BTX (benzene, toluene and xylene: 235μg/m3 in total) or acetone (133μg/m3). The geometric mean value of formaldehyde (19.2μg/m3) exceeded an 8-hour regulatory limit in Canada (9μg/m3), while those in other compounds did not exceed any regulatory limits, although a few residences exceeded at least one regulatory limit of benzene or acetaldehyde. Thus, the VOCs in the private residences were effectively characterized from the limited number of monitoring, and the potential health risks of the VOCs exposure, particularly formaldehyde, should be considered in the study area.

Effect of surfactants on the aggregation and stability of TiO2 nanomaterial in environmental aqueous matrices

The high likelihood of the coexistence of surfactants and engineered nano-materials (ENMs) in aqueous environments redoubles the need to study the effects of surfactants on the behavior and transport of nano-materials. In this study, we chose anionic sodium dodecyl sulfate (SDS) and nonionic nonylphenol ethoxylate (NPEO, Tergitol NP-9) to study the effect of surfactants on the behaviors of a typical TiO2 ENM in water environment. An adsorption isotherm and zeta potential study showed that both surfactants could be adsorbed onto the surface of nano-TiO2 but that only SDS can significantly decrease the zeta potential of TiO2. For a better simulation of the dynamic natural water environment, we conducted a 24-h batch experiment to study the aggregation behaviors of nano-TiO2 in the presence of SDS or NP-9 surfactants and recorded the sedimentation of the aggregates in solutions containing the surfactants over a 24h-period. SDS and NP-9 both reduced the growth of the TiO2 aggregates and retarded the aggregate sedimentation at surfactant concentrations ≥0.015% (w:v). The aggregation of TiO2 was far more remarkable in six environmental water matrices from different natural origins than in Milli-Q water, mainly because of the relatively high ionic strength of the former. The introduction of surfactants, however, still retarded the aggregation and sedimentation of TiO2 in the six environmental water matrices. SDS exerted a stronger reductive effect than NP-9. This study reveals that the presence of surfactants may increase the bioavailability and environmental risk of nanomaterial contaminants by retarding the aggregation and promoting the transport of nanomaterials in natural aqueous environments.

Functional-dependent and size-dependent uptake of nanoparticles in PC12

It is suggested that the uptake of nanoparticles is changed by the particle size or the surface modification. In this study, we quantified the uptake of nanoparticles in PC12 cells exposed Quantum Dots with different surface modification or fluorescent polystyrene particles with different particle size. The PC12 cells were exposed three types of the Quantum Dots (carboxyl base-functionalized, amino base-functionalized or non-base-functionalized) or three types of the fluorescent particles (22 nm, 100 nm or 1000 nm) for 3 hours. The uptake of the nanoparticles was quantified with a spectrofluorophotometer. The carboxyl base-functionalized Quantum Dots were considerably taken up by the cells than the non-base-functionalized Quantum Dots. Conversely, the amino base-functionalized Quantum Dots were taken up by the cells less frequently than the non-base-functionalized Quantum Dots. The particle number of the 22 nm-nanoparticles taken up by the cells was about 53 times higher than the 100 nm-particles. However, the particle weight of the 100 nm-particles taken up by the cells was higher than that of the 22 nm-nanoparticles. The 1000 nm-particles were adhered to the cell membrane, but they were little taken up by the cells. We concluded that nanoparticles can be taken up nerve cells in functional-dependent and size-dependent manners.

Bioaccessibility of Fukushima‐Accident‐Derived Cs in Soils and the Contribution of Soil Ingestion to Radiation Doses in Children

Ingestion of contaminated soil is one potential internal exposure pathway in areas contaminated by the Fukushima Daiichi Nuclear Power Plant accident. Doses from this pathway can be overestimated if the availability of radioactive nuclides in soils for the gastrointestinal tract is not considered. The concept of bioaccessibility has been adopted to evaluate this availability based on in vitro tests. This study evaluated the bioaccessibility of radioactive cesium from soils via the physiologically-based extraction test (PBET) and the extractability of those via an extraction test with 1 mol/L of hydrochloric acid (HCl). The bioaccessibility obtained in the PBET was 5.3% ± 1%, and the extractability in the tests with HCl was 16% ± 3%. The bioaccessibility was strongly correlated with the extractability. This result indicates the possibility that the extractability in HCl can be used as a good predictor of the bioaccessibility with PBET. In addition, we assessed the doses to children from the ingestion of soil via hand-to-mouth activity based on our PBET results using a probabilistic approach considering the spatial distribution of radioactive cesium in Date City in Fukushima Prefecture and the interindividual differences in the surveyed amounts of soil ingestion in Japan. The results of this assessment indicate that even if children were to routinely ingest a large amount of soil with relatively high contamination, the radiation doses from this pathway are negligible compared with doses from external exposure owing to deposited radionuclides in Fukushima Prefecture.

Aggregation is a critical cause of poor transfer into the brain tissue of intravenously administered cationic PAMAM dendrimer nanoparticles

Dendrimers have been expected as excellent nanodevices for brain medication. An amine-terminated polyamidoamine dendrimer (PD), an unmodified plain type of PD, has the obvious disadvantage of cytotoxicity, but still serves as an attractive molecule because it easily adheres to the cell surface, facilitating easy cellular uptake. Single-photon emission computed tomographic imaging of a mouse following intravenous injection of a radiolabeled PD failed to reveal any signal in the intracranial region. Furthermore, examination of the permeability of PD particles across the blood–brain barrier (BBB) in vitro using a commercially available kit revealed poor permeability of the nanoparticles, which was suppressed by an inhibitor of caveolae-mediated endocytosis, but not by an inhibitor of macropinocytosis. Physicochemical analysis of the PD revealed that cationic PDs are likely to aggregate promptly upon mixing with body fluids and that this prompt aggregation is probably driven by non-Derjaguin–Landau– Verwey–Overbeek attractive forces originating from the surrounding divalent ions. Atomic force microscopy observation of a freshly cleaved mica plate soaked in dendrimer suspension (culture media) confirmed prompt aggregation. Our study revealed poor transfer of intravenously administered cationic PDs into the intracranial nervous tissue, and the results of our analysis suggested that this was largely attributable to the reduced BBB permeability arising from the propensity of the particles to promptly aggregate upon mixing with body fluids.

Cost-Effective Strategy for the Investigation and Remediation of Polluted Soil Using Geostatistics and a Genetic Algorithm Approach

The geostatistical technique of Kriging has extensively been used for the investigation and delineation of soil heavy metal pollution. Kriging is rarely used in practical circumstances, however, because the parameter values are difficult to decide and relatively optimal locations for further sampling are difficult to find. In this study, we used large numbers of assumed actual polluted fields (AAPFs) randomly generated by unconditional simulation (US) to assess the adjusted total fee (ATF), an assessment standard developed for balancing the correct treatment rate (CTR) and total fee (TF), based on a traditional strategy of systematic (or uniform) grid sampling (SGS) and Kriging. We found that a strategy using both SGS and Kriging was more cost-effective than a strategy using only SGS. Next, we used a genetic algorithm (GA) approach to find optimal locations for the additional sampling. We found that the optimized locations for the additional sampling were at the joint districts of polluted areas and unpolluted areas, where abundant SGS data appeared near the threshold value. This strategy was less helpful, however, when the pollution of polluted fields showed no spatial correlation.