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The razor clam Sinonovacula constricta is a commercially important bivalve in Japan. The current distribution of this species in Japan is limited to Ariake Bay, where the fishery stock is declining. It is necessary to understand the genetic population structure of this species in order to restore the fishery stock while preserving the genetic diversity of the clam. Here, we report for the first time the genetic population structure of S. constricta in Ariake Bay, Japan. Paired-end restriction site-associated DNA sequencing (RAD-Seq) analyzed samples of S. constricta collected from seven mudflats located along Ariake Bay. Two different genetic populations exist in Ariake Bay, one inhabiting wild habitats and the other inhabiting the transplanted area of artificial seedlings. Our results suggest that genetic differentiation occurred between these two populations ( F st value = 0.052), and a high level of genetic differentiation is maintained between the two groups. In the future, monitoring the interbreeding status of the two genetically distinct populations and the genetic differentiation within each population is important for conserving the genetic diversity of S. constricta in Japan.

In the year 2018, black-spot shell disease, causing black discoloration of both the shell and pearls, spread widely and significantly impacted the cultured pearl industry. According to our previous study, black-spot shell disease has been attributed to the infection caused by Tenacibaculum sp. Pbs-1. To detect this bacterium specifically, we developed a specific quantitative polymerase chain reaction (qPCR) assay for Tenacibaculum sp. Pbs-1 in the present study. A specific primer set targeted for the internal spacer region (ISR) was designed. Thereafter, the specificity of the primer set was examined with strains of the genus Tenacibaculum . As a result of the optimization of the reaction conditions for qPCR, the original assays we developed were found to have high linearity and quantitative correlations (coefficient of 0.998) for standard plasmid DNA. The assay was able to quantify the strain Pbs-1 when only ten copies of the target ISR were present. Furthermore, we found that the quantification of the strain Pbs-1 was successfully conducted in both the shells of infected pearl oysters and natural seawater from the surrounding environment. These results indicate that our developed quantification method will contribute to clarifying the infection mechanism and assessing the severity of black-spot shell disease.

Two experiments tested a total of 509 participants on insight problems (the radiation problem and the nine-dot problem). Half of the participants were first exposed to a 1-min movie that included a subliminal hint. The hint raised the solution rate of people who did not recognize it. In addition, the way they solved the problem was affected by the hint. In Experiment 3 , a novel technique was introduced to address some methodological concerns raised by Experiments 1 and 2 . A total of 80 participants solved the 10-coin problem, and half of them were exposed to a subliminal hint. The hint facilitated solving the problem, and it shortened the solution time. Some implications of subliminal priming for research on and theorizing about insight problem solving are discussed.

Coastal hypoxia is an increasing environmental concern affecting marine ecosystems globally, particularly impacting benthic organisms such as bivalves. Although previous studies focused on the physiological responses of bivalves to hypoxic stress, the role of resident bacteria in the host response to hypoxia remains poorly understood. This study investigated changes in the resident bacterial communities in the gills and hepatopancreatic tissues of the ark shell ( Anadara kagoshimensis ) under hypoxic conditions. Specimens were assigned to three treatment groups: untreated control, hypoxia, and hypoxia with chloramphenicol supplementation (5.0 mg/L). After 3 days, specimens exposed to hypoxia exhibited black precipitation in the culture water, whereas antibiotic treatment reduced these effects. Amplicon sequencing revealed distinct bacterial communities between the tissues, with Arcobacteraceae and Alkalispirochaetaceae dominating in the gills and Metamycoplasmataceae being predominant in the hepatopancreas. The hepatopancreas displayed greater bacterial community changes than the gills under hypoxic conditions, including an increase in the abundance of Metamycoplasmataceae. The predicted metabolic functions suggested that these bacteria contribute to iron sulfide precipitation through sulfate reduction and iron respiration. The antibiotic-treated group displayed bacterial communities more similar to those of the control group, confirming the effectiveness of chloramphenicol in suppressing bacterial changes under hypoxia. This study provided new insights into tissue-specific bacterial responses to hypoxia in A . kagoshimensis and highlighted the potential role of Metamycoplasmataceae in the bivalve’s response to hypoxic stress.

In the inner part of Ariake Bay, located on the west coast of Kyushu, in western Japan, red tides have occurred with more frequency since the second half of the 1990s. Hypoxic waters have occurred during the summer months since the 2000s, despite the fact that nutrient loading from the land to the bay has not increased over the last five decades. We monitored water conditions at nine stations in the inner part of the bay, conducted benthic environmental surveys, and quantitative samplings of macro-benthic communities at the innermost four stations between 2002 and 2008. Each summer, the water was well-stratified due to the development of a halocline and a thermocline. The DO of the water below the pycnocline fell to hypoxic conditions. At the innermost three stations in the bay, the mud content and organic matter content of the sediment increased significantly, and the carbon stable isotope ratios of the organic matter contained in the sediment ranged between −21.3±0.5‰ and −20.7±0.5‰ of δ13C. These facts indicated that the organic matter was derived photosynthetically from marine phytoplankton. The increase in the mud content of the sediment indicates a deceleration in the tidal current. This may be a key event that induces a series of environmental changes and disturbances, including the stratification of the water, the more frequent occurrence of red tides, the progress of the organic enrichment of the sediment, and the occurrence of hypoxic water during the summer.

The objective of this study was to determine the effects of transported hypoxic water on macrobenthic species composition in Isahaya Bay, Kyushu, Japan. To this end, we conducted a field survey of water quality, sediment quality, and the macrobenthic community across seasons. Hypoxic intensity, defined as the percentage of the time that a region exhibits hypoxic conditions [dissolved oxygen (DO) < 3.0 mg L −1 ], was calculated for the summer period from July 1 to September 30, 2014 based on continuous monitoring data derived from six monitoring towers in the bay. Here, we discuss how the hypoxia-induced damage, defined as the change in the species compositions of the macrobenthic communities in the bay due to hypoxia, differs between the inner and middle regions of the bay (with muddy sediment) and the outer regions of the bay (with sandy sediment). A decrease in the density of macrobenthic species that was correlated to the period of lowest DO concentration (< 1.0 mg L −1 ) was observed in the inner region of the bay. A large number of species disappeared from the macrobenthic community in the outer region after the DO concentration had dropped to its lowest level, even though this region presented the lowest hypoxic intensity (6.6%). The species that disappeared, including crustaceans and some polychaetes, were presumed to be sensitive to, and to have escaped from, hypoxia. Fewer species disappeared from the inner and intermediate regions of the bay, which had high hypoxic intensities of 14.0 and 26.7%, respectively, than from the outer region. The reason for this appeared to be a predominance of hypoxia-tolerant species in the inner and intermediate regions but not in the outer region. Our study suggests that estimating the effect of hypoxia in a spatially heterogeneous environment must be done with caution.

Abstract The razor clam Sinonovacula constricta is a commercially important bivalve in Japan. The current distribution of this species in Japan is limited to Ariake Bay, where the fishery stock is declining. It is necessary to understand the genetic population structure of this species in order to restore the fishery stock while preserving the genetic diversity of the clam. Here, we report for the first time the genetic population structure of S. constricta in Ariake Bay, Japan. Paired-end restriction site-associated DNA sequencing (RAD-Seq) analyzed samples of S. constricta collected from seven mudflats located along Ariake Bay. Two different genetic populations exist in Ariake Bay, one inhabiting wild habitats and the other inhabiting the transplanted area of artificial seedlings. Our results suggest that genetic differentiation occurred between these two populations (Fst value = 0.052), and a high level of genetic differentiation is maintained between the two groups. In the future, monitoring the interbreeding status of the two genetically distinct populations and the genetic differentiation within each population is important for conserving the genetic diversity of S. constricta in Japan. Competing Interest Statement The authors have declared no competing interest.

Post-transcriptional modifications have critical roles in tRNA stability and function 1 – 4 . In thermophiles, tRNAs are heavily modified to maintain their thermal stability under extreme growth temperatures 5 , 6 . Here we identified 2′-phosphouridine (U p ) at position 47 of tRNAs from thermophilic archaea. U p 47 confers thermal stability and nuclease resistance to tRNAs. Atomic structures of native archaeal tRNA showed a unique metastable core structure stabilized by U p 47. The 2′-phosphate of U p 47 protrudes from the tRNA core and prevents backbone rotation during thermal denaturation. In addition, we identified the arkI gene, which encodes an archaeal RNA kinase responsible for U p 47 formation. Structural studies showed that ArkI has a non-canonical kinase motif surrounded by a positively charged patch for tRNA binding. A knockout strain of arkI grew slowly at high temperatures and exhibited a synthetic growth defect when a second tRNA-modifying enzyme was depleted. We also identified an archaeal homologue of KptA as an eraser that efficiently dephosphorylates U p 47 in vitro and in vivo. Taken together, our findings show that U p 47 is a reversible RNA modification mediated by ArkI and KptA that fine-tunes the structural rigidity of tRNAs under extreme environmental conditions. Reversible internal RNA phosphrylation contributes to thermal stability and nuclease resistance of tRNA, and cellular thermotolerance of hyperthermophiles. 

Abstract BACKGROUND The hemodynamic changes after superficial temporal artery (STA) to middle cerebral artery (MCA) bypass surgery are unclear. OBJECTIVE To clarify the hemodynamics by comparing flow parameters obtained by 4-dimensional (4D) flow magnetic resonance imaging (MRI) and intraoperative MCA pressure measurement. METHODS We recruited 23 patients who underwent STA-MCA bypass surgery for internal carotid artery (ICA) or MCA stenosis. We monitored intraoperative MCA, STA, and radial artery (RA) pressure. All patients underwent 4D flow MRI preoperatively and 3 wk after surgery to quantify the blood flow volume (BFV) of the ipsilateral ICA (BFViICA), contralateral ICA (BFVcICA), basilar artery (BFVBA), ipsilateral STA (BFViSTA), and contralateral STA (BFVcSTA). The sum of intracranial BFV was defined as BFVtotal. We compared BFV parameters and intraoperative pressure. RESULTS BFViSTA significantly increased after surgery (P < .001). BFViICA and BFVBA significantly decreased after surgery (BFViICAP = .005; BFVBAP = .02). No significant difference was observed between BFVcICA before and after surgery. As a result, BFVtotal postoperatively increased by 6.8%; however, no significant difference was observed. Flow direction at M1 changed from antegrade to unclear after surgery in 5 patients. Intraoperative MCA pressure and MCA/RA pressure ratio significantly increased after surgery (P < .001). We found a stronger positive correlation between MCA pressure increase ratio and BFVtotal increase ratio in patients with lower pre-MCA pressure (r = 0.907, P < .001). CONCLUSION The visual and quantitative assessment of 4D flow MRI revealed that intracranial blood flow changes complementarily after STA-MCA bypass surgery. 4D flow MRI may detect the improvement of cerebral perfusion pressure.

Background We evaluated the prevalence and identified the risk factors for retropharyngeal and retro-styloid lymph node metastasis (LNM) in patients with hypopharyngeal carcinoma (HPC). This was achieved using a combination of magnetic resonance (MR) and [ 18  F]-fluoro-2-deoxy-D-glucose (FDG)–positron emission tomography (PET)/computed tomography (CT) images. Methods Two board-certified radiation oncologists retrospectively reviewed pretreatment FDG–PET/CT images and contrast-enhanced thin-slice CT and MR images of 155 patients with HPC who underwent radiotherapy. Fisher’s exact tests and logistic regression analyses were performed to assess the risk factors for LNM. Results Retropharyngeal LNM (RPLNM) was confirmed in 20 (13%) patients. Posterior wall (PW) tumors (odds ratio [OR]: 4.128, 95% confidence interval [CI]: 1.339–12.727; p  = 0.014) and bilateral or contralateral cervical LNM (OR: 11.577, 95% CI: 2.135–62.789; p  = 0.005) were significantly correlated with RPLNM. The RPLNM was found in 9 (32%) of the 28 patients with PW tumors. Of these 9 patients, 2 (7%) had ipsilateral RPLNM, 3 (11%) had contralateral RPLNM, and 4 (14%) had bilateral RPLNM. The PW tumors were significantly associated with contralateral RPLNM ( p  < 0.001). Retro-styloid LNM (RSLNM) was confirmed in two (1%) patients, both of whom had ipsilateral RSLNM with lymph nodes (LNs) of ≥ 15 mm in the upper limit of ipsilateral level II. A significant association was found between LNs of ≥ 15 mm in the upper limit of ipsilateral level II and ipsilateral RSLNM ( p  = 0.001). Conclusions The RPLNM was identified in 13% of patients with HPC. The PW tumors and bilateral or contralateral cervical LNM were risk factors for RPLNM; particularly, PW tumors were a specific risk factor for contralateral RPLNM. Although the RSLNM was rare, LNs of ≥ 15 mm in the upper limit of ipsilateral level II were a risk factor for ipsilateral RSLNM.