A case of paralytic shellfish poisoning caused by consumption of visceral balls from geoduck Panopea japonica in Japan.

In the end of March 2018, an unprecedented food poisoning incident due to ingestion of the visceral balls of geoduck Panopea japonica occurred in Japan. The patient, presented with symptoms of numbness on the lips and general weakness, was diagnosed as paralytic shellfish poisoning (PSP). The patient immediately treated with the mechanical ventilation recovered and left the hospital after 3 days treatment. Saxitoxins (STXs) in the plasma and urinary samples collected from the patient on the first and second day after hospitalization were analyzed by ultra high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC/MS/MS) and liquid chromatography with post-column fluorescent detection (LC/FLD). The STXs levels of 499.1 and 6.0 µg/L of STX dihydrochloride equivalent (STX·2HCl eq.) were quantitated by LC/FLD in the urinary samples on the first and second day, respectively. In addition, geoducks harvested from the same areas of the PSP causative specimens after the incident were analyzed by LC/FLD, and the results showed the level of STXs in their whole bodies of the geoducks exceeding 0.8 mg STX·2HCl eq./kg which is the maximum levels of STX in CODEX STAN 292-2008. Prominent toxins in STXs that detected in urinary and geoduck samples and identified by UHPLC/MS/MS and LC/FLD were gonyautoxin-1+4 (GTX1+4). These results concluded that the incident was the food poisoning due to STXs accumulated in the geoducks. This is the first PSP case caused by consumption of geoducks in Japan. This is also the first PSP case that causative toxins are detected in urinary samples of patients involved in PSP in Japan.

New azaspiracid analogues detected as bi-charged ions in Azadinium poporum (Amphidomataceae, Dinophyceae) isolated from Japanese coastal waters.

Lipophilic marine biotoxin azaspiracids (AZAs) are produced by dinoflagellates Azadinium and Amphidoma. Recently, several strains of Azadinium poporum were isolated from Japanese coastal waters, and detailed toxin profiles of two strains (mdd421 and HM536) among them were clarified by several detection techniques on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOFMS). In our present study, AZA analogues in seven strains of A. poporum from Japanese coastal waters (including two previously reported strains) were determined by these detection techniques. The dominant AZA in the seven strains was AZA2 accompanied by small amounts of several known AZAs and twelve new AZA analogues. Eight of the twelve new AZA analogues discovered in our present study were detected as bi-charged ions on the positive mode LC/MS/MS. This is the first report describing AZA analogues detected as bi-charged ions with hexose and sulfate groups in their structures.

Azaspiracid accumulation in Japanese coastal bivalves and ascidians fed with Azadinium poporum producing azaspiracid-2 as the dominant toxin component.

The filter-feeding bivalves often accumulate marine toxins by feeding on toxic dinoflagellates that produce marine toxins. Azaspiracids (AZAs) are a group of lipophilic polyether toxins which have been detected in a variety of organisms in many countries. In our present study, accumulation kinetics and toxin distributions in the tissues of seven bivalve species and ascidians relevant to Japanese coastal waters were investigated by experimentally feeding a toxic dinoflagellate Azadinium poporum, which produces azaspiracid-2 (AZA2) as the dominant toxin component. All bivalve species and ascidians investigated in this study had the capability to accumulate AZA2 and no metabolites of AZA2 were detected in the bivalves and the ascidians. Japanese short-neck clams, Japanese oysters, Pacific oysters and ascidians accumulated AZA2 with the highest concentrations on the hepatopancreas, whereas the highest concentrations of AZA2 were found on the gills in surf clams and horse clams. Hard clams and cockles accumulated high levels of AZA2 in both the hepatopancreas and the gills. As far as we know, this is the first report describing detailed tissue distribution of AZAs in several bivalve species other than mussels (M. edulis) and scallops (P. maximus). Variation of accumulation rates of AZA2 in Japanese short-neck clams on different cell densities or temperatures were observed.

Coherent Doppler LIDAR with long-duration frequency-modulated pulses for wind sensing.

A coherent Doppler LIDAR (CDL) with long-duration frequency-modulated pulses was demonstrated and validated by analyzing the data observed by a prototype. In traditional CDL using short-duration single-frequency pulses (PCDL; pulsed CDL), there exists a trade-off relationship between distance and velocity resolution. Meanwhile, in earlier work, a theoretical framework of CDL using long-duration frequency-modulated pulses (FMCDL; frequency-modulated CDL) was put forth to eliminate the trade-off. We developed the prototype to be operated as both a PCDL and FMCDL. Analyses of data observed by the PCDL and FMCDL modes showed that the FMCDL worked in good agreement with the PCDL for wind ranging and velocimetry. Furthermore, the performance of the FMCDL in terms of received power and resolution of distance and velocity was quantitatively consistent with ones theoretically expected.

Unveiling the reaction pathways of hydrocarbons via experiments, computations and data science.

Reaction networks of hydrocarbons are explored using first principles calculations, data science, and experiments. Transforming hydrocarbon data into networks reveals the prevalence of the formation and reaction of various molecules. Graph theory is implemented to extract knowledge from the reaction network. In particular, centralities analysis reveals that H+, CCC, CH3+, CC, and [CH2+]C have high degrees and are thus very likely to form or react with other molecules. Additionally, H+, CH3+, C2H5+, C8H15+, C8H17+, and C6H11+ are found to have high control throughout the network and lead towards a series of additional reactions. The constructed network is also validated in experiments while the shortest path analysis is implemented for further comparison between experiment and the network. Thus, combining network analysis with first principles calculations uncovers key points in the development of various hydrocarbons that can be used to improve catalyst design and targeted synthesis of desired hydrocarbons.

Assessing the ADC of Bone-marrow on Whole-body MR Images in Relation to the Fat-suppression Method and Fat Content.

PURPOSE: To compare apparent diffusion coefficients (ADCs) of bone marrow on diffusion-weighted imaging (DWI) between two fat-suppression techniques, and to evaluate the association between bone-marrow ADCs and the proton density fat fraction (PDFF). METHODS: Seventy-seven patients underwent whole-body DWI with short-inversion time inversion-recovery (STIR) (DWISTIR) and/or STIR + selective water-excitation (spectral-spatial RF [SSRF]) (DWISTIR+SSRF). ADCs of lumbar vertebrae (L3 and L4) were compared between DWISTIR and DWISTIR+SSRF, and correlated with the PDFF. RESULTS: Lumbar ADCs obtained by DWISTIR and DWISTIR+SSRF were significantly correlated (L3: r = 0.90, P < 0.0001, L4: r = 0.90, P < 0.0001). Lumbar ADCs (× 10-6 mm2/s) obtained by DWISTIR were significantly lower than those by DWISTIR+SSRF (L3: 479 ± 137 and 490 ± 148, P < 0.05, L4: 456 ± 114 and 471 ± 118, P < 0.005). Residual fat signals were more clearly observed on DWISTIR than on DWISTIR+SSRF. The ADCs of L3 obtained by DWISTIR and DWISTIR+SSRF exhibited significant positive correlations with the PDFF (r = 0.51, P < 0.0001, and r = 0.45, P < 0.0001, respectively), and the ADCs of L4 obtained by DWISTIR and DWISTIR+SSRF exhibited significantly positive correlations with the PDFF (r = 0.40, P < 0.0005, and r = 0.40, P < 0.0005, respectively). CONCLUSION: Irrespective of different fat-suppression methods, lumbar ADCs were positively correlated with the PDFF, being inconsistent with previous studies. Lumbar ADCs obtained by DWISTIR were significantly lower than those obtained by DWISTIR+SSRF, probably due to residual fat signals on DWISTIR. However, this difference (< 4%) did not explain the positive correlation between lumbar ADC and PDFF.

[Rapid Screening System for Paralytic Shellfish Toxins in Bivalves by Oligonucleotide Lateral Flow Immunoassay].

The mouse bioassay (MBA) for paralytic shellfish toxins (PSTs) in bivalves has been used as an official method in Japan. It is necessary to develop an alternative method to animal experiments in PSTs assay because 3Rs (Replacement, Reduction, and Refinement) of animal experiments are required from the animal welfare point of view. Various methods such as HPLC-FL, receptor binding assay, LC-MS/MS and ELISA have been established to detect PSTs without performing animal experiments. The present study was undertaken to develop a screening method using oligonucleotide lateral flow immunoassay (OLFIA) for detecting PSTs in bivalves. The screening level was defined as positive at 2 MU/g of MBA that is the half regulation limit of PSTs monitoring in Japan. All 20 positive (equal to or more than 2 MU/g) samples judged from MBA showed a positive reaction in the OLFIA. No positive samples resulted in a false negative reaction. The OLFIA exhibited high accuracy at 2 MU/g of screening criteria. The authors demonstrated here that the OLFIA can be useful for rapid detection of PSTs in bivalves.

Complex profiles of azaspiracid analogues in two culture strains of Azadinium poporum (Amphidomataceae, Dinophyceae) isolated from Japanese coastal waters determined by LC-MS/MS.

Lipophilic marine biotoxins azaspiracids (AZAs) are produced by dinoflagellates Azadinium and Amphidoma. Recently, several strains of Azadinium poporum were isolated from Japanese coastal waters. In our present study, AZA analogues in two strains (mdd421 and HM536) of A. poporum were analyzed by several detection techniques on the liquid chromatography-tandem mass spectrometry (LC-MS/MS) and liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOFMS). The dominant AZA analogue in the Japanese A. poporum strains was AZA2. Other known AZA analogues were AZA11, AZA35, AZA2 methyl ester and AZA2 phosphate ester. Besides these AZAs, thirteen new AZA analogues were discovered in the two strains. A putative AZA analogue (Compound 1) with the smallest molecular weight ever found in nature was also discovered in the two strains. This is the first report describing detailed AZA profiles in Japanese isolates of A. poporum.

Development of an absolute quantification method for ribosomal RNA gene copy numbers per eukaryotic single cell by digital PCR.

Recent increase of Harmful Algal Blooms (HAB) causes world-wide ecological, economical, and health issues, and more attention is paid to frequent coastal monitoring for the early detection of HAB species to prevent or reduce such impacts. Use of molecular tools in addition to traditional microscopy-based observation has become one of the promising methodologies for coastal monitoring. However, as ribosomal RNA (rRNA) genes are commonly targeted in molecular studies, variability in the rRNA gene copy number within and between species must be considered to provide quantitative information in quantitative PCR (qPCR), digital PCR (dPCR), and metabarcoding analyses. Currently, this information is only available for a limited number of species. The present study utilized a dPCR technology to quantify copy numbers of rRNA genes per single cell in 16 phytoplankton species, the majority of which are toxin-producers, using a newly developed universal primer set accompanied by a labeled probe with a fluorophore and a double-quencher. In silico PCR using the newly developed primers allowed the detection of taxa from 8 supergroups, demonstrating universality and broad coverage of the primer set. Chelex buffer was found to be suitable for DNA extraction to obtain DNA fragments with suitable size to avoid underestimation of the copy numbers. The study successfully demonstrated the first comparison of absolute quantification of 18S rRNA copy numbers per cell from 16 phytoplankton species by the dPCR technology.

Determination of optimal culture conditions for toxin production by a Prorocentrum lima complex strain with high diarrhetic shellfish toxins yield.

Diarrhetic shellfish poisoning (DSP) is caused by the consumption of shellfish contaminated by diarrhetic shellfish toxins (DSTs) such as okadaic acid (OA) and dinophysistoxins (DTXs) produced by some species of dinoflagellates. To prevent the occurrence of human intoxication cases, inspection of DSTs (OA and DTXs) in shellfish is important. An instrumental method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been recently employed in Japan for the monitoring of OA and DTXs in shellfish. For such analysis, reference materials (RMs) of OA and DTXs are essential. Demand for the reference materials, especially dinophysistoxin-1 (DTX1), is recently increasing in Japan. Production of the materials has been performed by mass cultivation of a dinoflagellate (Prorocentrum lima) strain that produces DTXs and OA, which indicates that the efficiency of production depends on the toxin production of the strain used. In this study, P. lima complex subclade 1e strain MIO12P was determined to be a high DTX1 producer among the three Japanese strains of the P. lima complex (subclades 1e, 1f, and 1i). It was clarified that the culture medium suitable for toxin production by strain MIO12P was metals mix SWII medium, and the optimal temperature and salinity for toxin production were 25 °C and salinity 30, respectively. The DTX1 yield (1265.3 ng ml-1) of strain MIO12P cultured under the conditions described above was the highest reported worldwide. Prorocentrum lima complex subclade 1e strain MIO12P is expected to be useful for the sustainable production of DTX1 as a source of RMs for chemical and biochemical methods in the future.

SxtA localizes to chloroplasts and changes to its 3'UTR may reduce toxin biosynthesis in non-toxic Alexandrium catenella (Group I)✰.

SxtA is the enzyme that catalyses the first step of saxitoxin biosynthesis. We developed an immunofluorescent method to detect SxtA using antibodies against SxtA peptides. Confocal microscopy revealed the presence of abundant, sub-cellularly localized signal in cells of toxic species and its absence in non-toxic species. Co-localization of SxtA with Rubisco II and ultra-structural observation by transmission electron microscopy strongly suggested the association of SxtA with chloroplasts. We also characterized a non-toxic sub-clone of Alexandrium catenella (Group I) to elucidate the mutation responsible for its loss of toxicity. Although sxtA4 gene copy number was indistinguishable in toxic and non-toxic sub-clones, mRNA and protein expression were significantly reduced in the non-toxic sub-clone and we uncovered sequence variation at the 3' untranslated region (3'UTR) of sxtA4 mRNA. We propose that differences in the sxtA4 mRNA 3'UTR lead to down-regulation of STX biosynthesis post-transcriptionally, thereby explaining the differences in toxicity amongst different A. catenella (Group I) sub-clones.

Integrated [18F]FDG PET/MRI demonstrates the iron-related bone-marrow physiology.

We identified predictors for bone-marrow [18F]FDG uptake and MR signals among complete blood count, C-reactive protein (CRP), and anthropometric factors, and demonstrated the bone-marrow physiology using integrated [18F]FDG-PET/MRI. 174 oncology patients without bone-marrow lesions underwent whole-body [18F]FDG-PET/MRI. The standardized uptake value (SUV), apparent diffusion coefficient (ADC), proton density fat-fraction (PDFF), and a reciprocal of T2* relaxation time (R2*) were measured in lumbar vertebrae (L3-5) and bilateral ilia. Vertebrae, pelvis, and ribs were evaluated by 3-point visual scoring on DWI. The association of the PET/MR features with the predictors was examined. Multi-regression analyses identified CRP as the strongest predictor for lumbar and iliac SUVs (standardized coefficient: ß = 0.31 and ß = 0.38, respectively), and for lumbar and iliac R2* (ß = 0.31 and ß = 0.46, respectively). In contrast, age was the strongest factor influencing lumbar and iliac ADCs (ß = 0.23 and ß = 0.21, respectively), and lumbar and iliac PDFFs (ß = 0.53 and ß = 0.54, respectively). Regarding DWI-visual scores, age was the strongest predictor for vertebrae (ß = - 0.47), and the red cell distribution width (RDW) was the strongest predictor for pelvis and ribs (ß = 0.33 and ß = 0.47, respectively). The bone-marrow [18F]FDG uptake and R2* reflect anemia of inflammation (increased granulopoiesis and reduced iron metabolism), whereas bone-marrow DWI and PDFF reflect age and anemia-responsive erythropoiesis.

Abundance of the benthic dinoflagellate Prorocentrum and the diversity, distribution, and diarrhetic shellfish toxin production of Prorocentrum lima complex and P. caipirignum in Japan.

In the present study, the abundance of Prorocentrum and the molecular phylogeny, distribution, and DST production of P. lima complex and P. caipirignum in Japan were investigated. First, the cell densities of Prorocentrum were assessed from the temperate to subtropical zones in Japan between 2014 and 2018. The cell density in the subtropical zone [19.0 ± 40.2 cells/g wet weight (ww) algae] was significantly higher than that in the temperate zone (1.4 ± 3.4 cells/g ww algae). A total of 244 clonal strains were established from the temperate and subtropical zones. Phylogenetic analyses based on the large-subunit ribosomal DNA D1/D2 revealed that the strains were separated into four species/species complex/phylotypes (P. lima complex, P. caipirignum, and new phylotypes Prorocentrum spp. types 1 and 2). The strains of P. lima complex could be separated into two clades (1 and 3). Furthermore, the strains of clades 1 and 3 could be separated into nine subclades (1a, 1c, 1d, 1e, 1f, 1g, 1h, 1i, and 1j) and three subclades (3a, 3b, and 3c), respectively. The strains of P. caipirignum were separated into two subclades (b and e). Each phylotype/subclade showed a unique distribution pattern in Japan: P. lima complex subclades 1a, 1c, and 3a and P. caipirignum subclades b and e were widespread from the temperate to subtropical zones. On the other hand, P. lima complex subclades 1e and 1i were restricted to the temperate zone, and P. lima complex subclades 1d, 1f, 1g, 1h, 1j, 3b, and 3c and Prorocentrum spp. types 1 and 2 were restricted to the subtropical zone. Furthermore, the DST production of the 243 clonal strains was assessed by LC/MS/MS analysis. The results revealed that all strains produced okadaic acid (OA) and that the OA contents of P. lima complex subclades 1d and 1f, P. caipirignum subclades b and e, and Prorocentrum sp. type 2 tended to be higher than those of the other subclades. While P. lima complex subclades 1a, 1e, 1f, and 1i produced DTX1, the other phylotype/subclades produced either no or low quantities of DTX1. A strain of P. lima complex subclade 1e showed the highest OA and DTX1 contents (55.27 and 70.73 pg/cell, respectively) in the world. These results suggest that there are potential risks for DST accumulation in benthic animals in Japan.

Zero Echo Time-Based PET/MRI Attenuation Correction in Patients With Oral Cavity Cancer: Initial Experience.

PURPOSE: The aims of this study were to demonstrate the feasibility of zero echo time (ZTE) MRI for jawbone identification, and to evaluate the quantitative performance of F-FDG PET/MRI with ZTE-based attenuation correction (ZTE-AC) compared with PET/CT and PET/MRI with Dixon MR-based AC (Dixon-AC) in patients with oral cavity cancer (OCC). MATERIALS AND METHODS: Thirteen OCC patients underwent whole-body F-FDG PET/CT and subsequent regional PET/MRI with Dixon-AC and ZTE-AC in 1 day. SUVs of the primary OCC and metastatic cervical lymph nodes (CLNs) were measured on PET/CT (SUVCT), PET/MRI with Dixon-AC (SUVDixon), and ZTE-AC (SUVZTE). The SUVs were then compared. RESULTS: The ZTE MRI scans minimized the effects of metal artifacts from dentures, and ZTE-AC maps correctly delineated the jawbones. SUVDixon and SUVZTE had significant positive correlations with SUVCT (Pearson r = 0.97 and r = 0.99 for OCC, and r = 0.98 and r = 0.98 for CLNs, respectively). The mean ± SD of SUVCT, SUVDixon, and SUVZTE were 14.4 ± 8.0, 14.5 ± 8.6, and 15.6 ± 8.8 for OCC, and 6.3 ± 3.0, 8.0 ± 4.0, and 7.6 ± 3.9 for CLNs, respectively. For OCCs, SUVZTE was significantly higher than SUVCT (P < 0.05), whereas there was no significant difference between SUVCT and SUVDixon or between SUVDixon and SUVZTE. For CLNs, SUVDixon and SUVZTE were significantly higher than SUVCT (P < 0.01 and P < 0.05, respectively), and SUVDixon was significantly higher than SUVZTE (P < 0.01). CONCLUSIONS: ZTE MRI can correctly identify jawbones while minimizing the effects of metal artifacts. The ZTE-AC method in F-FDG PET/MRI reduces the underestimation of tracer uptake due to Dixon-AC jawbone errors and improves the quantitative performance of PET for OCC patients.

Heptavalinamide A, an Extensively N-Methylated Linear Nonapeptide from a Cyanobacterium Symploca sp. and Development of a Highly Sensitive Analysis of N,N-Dimethylvaline by LCMS.

An extensively N-methylated linear nonapeptide heptavalinamide A (1) was isolated from the marine cyanobacterium Symploca sp. collected at Kabira Reef of Ishigaki Island, Okinawa. The amino acid sequence of 1 was assigned by interpretation of 2D NMR and MS/MS data. The absolute configurations of the constituent amino acids were determined by the application of Marfey's method. A method to assign the configuration of N,N-dimethylvaline by LCMS is discussed.

Monitoring of the toxic dinoflagellate Alexandrium catenella in Osaka Bay, Japan using a massively parallel sequencing (MPS)-based technique.

Since 2002, blooms of Alexandrium catenella sensu Fraga et al. (2015) and paralytic shellfish toxicity events have occurred almost yearly in Osaka Bay, Japan. To better understand the triggers for reoccurring A. catenella blooms in Osaka Bay, phytoplankton community was monitored during the spring seasons of 2012-2015. Monitoring was performed using massively parallel sequencing (MPS)-based technique on amplicon sequences of the 18S rRNA gene. Dense blooms of A. catenella occurred every year except in 2012, however, there was no significant correlation with the environmental parameters investigated. Plankton community diversity decreased before and middle of the A. catenella blooms, suggesting that the decline in diversity could be an indicator for the bloom occurrence. The yearly abundance pattern of A. catenella cells obtained by morphology-based counting coincided with the relative sequence abundances, which supports the effectiveness of MPS-based phytoplankton monitoring.

Quantitation of Tetrodotoxin and Its Analogues with a Combination of Liquid Chromatography-Tandem Mass Spectrometry and Quantitative 1H-NMR Spectroscopy.

Tetrodotoxin and its analogues are the causative toxins of pufferfish poisoning. Tetrodotoxin has been recently detected in bivalve mollusks collected in New Zealand and Europe, highlighting the need to include tetrodotoxin in monitoring programs for bivalves by instrumental methods. In the present study, tetrodotoxin and its analogues in commercially available tetrodotoxin reagents were quantitated accurately by quantitative 1H-nuclear magnetic resonance (qNMR) spectroscopy. The results were applied to estimate relative molar responses of tetrodotoxin and its analogues in hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC/MS/MS). All four components (tetrodotoxin hemilactal form (1), tetrodotoxin 10,7-lactone form (2), 4-epitetrodotoxin (3), and 4,9-anhydrotetrodotoxin (4)) generated by equilibrating tetrodotoxin in aqueous solution were prepared as a mixture. From the HSQC spectrum of the mixture, the separated signals derived from three components, excluding 1, were selected and used for the quantitation. In addition, the relative molar responses of 3 and 4 on HILIC/MS/MS were calculated to be 0.73 and 0.46, respectively. These values could be useful for quantitation of 3 and 4 using the tetrodotoxin standard by HILIC/MS/MS. Our results also indicate that qNMR is useful for preparation of tetrodotoxin certified reference material.

Development of Ultra-Performance Liquid Chromatography with Post-Column Fluorescent Derivatization for the Rapid Detection of Saxitoxin Analogues and Analysis of Bivalve Monitoring Samples.

Saxitoxin (STX) and its analogues produced by toxic dinoflagellates accumulate in bivalves, and routine monitoring of bivalves is important to prevent cases of human poisoning. In this study, we describe a rapid detection method for the analysis of STXs using ultra-performance liquid chromatography with post-column fluorescent detection and to investigate water depths and sampling points optimal for shellfish toxin monitoring. Cultured scallops (Mizuhopecten yessoensis) and mussels (Mytilus galloprovincialis) were collected from various water depths and sampling points were used in this study. Irrespective of bivalve species, toxin concentrations in bivalves were lower at deeper water depths. The toxin concentrations of bivalves did not differ greatly when bivalves were collected from the same bay. Although the levels of contamination of bivalves with STXs can depend on various environmental and geographical factors, our findings are useful for formulating a sampling protocol for the prevention of harvesting contaminated shellfish.

Whole-body bone marrow DWI correlates with age, anemia, and hematopoietic activity.

PURPOSE: To identify the predictors of bone-marrow DWI signals from anthropometric, complete blood count (CBC), and C-reactive protein (CRP), and to evaluate the association with fat-content in patients. METHOD: This retrospective study was approved by the institutional review board. A total of 113 consecutive tumor patients underwent whole-body PET/MRI. Apparent diffusion coefficient (ADC) and proton density fat fraction (PDFF) were measured and averaged in lumbar vertebrae (L3-5) and bilateral ilia. Due to respiratory motion, ribs were evaluated by 3-point visual scoring on DWI with b = 800 (1: invisible, 2: partially visible, 3: fully visible). The relationships between ADC/visual scores and anthropometric, CBC, CRP, and PDFF were examined. In females, the age-dependency was evaluated. RESULTS: Multi-regression analyses identified age as the strongest predictor of lumbar ADC (standardized coefficient: ß = 0.45), followed by red cell distribution width (RDW) (ß = -0.24), while age was the strongest predictor of iliac ADC (ß = 0.43), followed by hemoglobin (Hb) (ß = 0.22). RDW was the strongest predictor (ß = 0.47) for rib visual score and age was the second (ß = -0.39). ADC showed significant positive correlations with PDFF at L3-5 and ilium. Lumbar ADC showed a decreasing trend during middle age in females. CONCLUSIONS: Age, anemia (lower Hb), and increased hematopoietic activity (higher RDW) are the predominant predictors of ADC and the visibility of red marrow on DWI. Fat-suppression methods and bone-marrow physiology in middle-aged females may have affected the measured correlations between ADC and PDFF inconsistent with previous studies.

Genetic relatedness of a new Japanese isolates of Alexandrium ostenfeldii bloom population with global isolates.

In recent years, blooms of toxic Alexandrium ostenfeldii strains have been reported from around the world. In 2013, the species formed a red tide in a shallow lagoon in western Japan, which was the first report of the species in the area. To investigate the genetic relatedness of Japanese A. ostenfeldii and global isolates, the full-length SSU, ITS and LSU sequences were determined, and phylogenetic analyses were conducted for isolates from western and northern Japan and from the Baltic Sea. Genotyping and microsatellite sequence comparison were performed to estimate the divergence and connectivity between the populations from western Japan and the Baltic Sea. In all phylogenetic analyses, the isolates from western Japan grouped together with global isolates from shallow and low saline areas, such as the Baltic Sea, estuaries on the east coast of U.S.A. and from the Bohai Sea, China. In contrast, the isolates from northern Japan formed a well-supported separate group in the ITS and LSU phylogenies, indicating differentiation between the Japanese populations. This was further supported by the notable differentiation between the sequences of western and northern Japanese isolates, whereas the lowest differentiation was found between the western Japanese and Chinese isolates. Microsatellite genotyping revealed low genetic diversity in the western Japanese population, possibly explained by a recent introduction to the lagoon from where it was detected. The red tide recorded in the shallow lagoon followed notable changes in the salinity of the waterbody and phytoplankton composition, potentially facilitating the bloom of A. ostenfeldii.