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Very strong intra- and inter-molecular hydrogen-bonding interactions coexist with strong π–π overlaps in a pure organic superconductor (TEA)(HEDO-TTF-dc)2·2(H2C2O4).

Nicotinamide adenine dinucleotide (NAD+) is a coenzyme essential for energy production. Recently, associations between NAD+ and aging-related diseases have been reported, and NAD+ precursors that increase NAD+ concentration in the body have been acknowledged as anti-aging supplements. However, there have been only a few studies on the link between aging or aging-related diseases and human blood NAD+ concentration because NAD+ and its precursors are unstable in blood and difficult to measure. Therefore, we aimed to construct a quantitative NAD+ measurement method that is simpler than the existing methods. The calibration standards of NAD+ showed good linearity (0.9936 to 0.9990) in the range of 0.25 to 200 μM, and the lower limit of quantification was 0.5 to 2 μM. We found that QIAcard FTA DMPK-B maintained NAD+ stability of 85% or more for at least 2 weeks at 4 °C and 1 week at room temperature using the dried blood spot method. Additionally, NAD+ stability in the blood extraction solution was more than 90% for 2 months. To our knowledge, there has been no report on a quantitative NAD+ measurement method in human whole blood that can be performed with as little as 5 μL of blood and can be easily implemented at both medical clinics and private homes. Our simple and convenient method has the potential to become the gold standard for NAD+ measurement in blood. It is expected to contribute to the acceleration of research on the correlation between aging or aging-related diseases and NAD+ concentration in human blood.

Background: This study aimed to conduct a preliminary evaluation of the effects of 6-methylsulfinylhexel isothiocyanate (6-MSITC) contained in wasabi rhizomes on brain fatigue and sleep and examine its safety through overdose study. Methods: A total of 20 healthy volunteers who were experiencing daily fatigue were given wasabi extract powder containing 6-MSITC (4.8 mg/day of 6-MSITC) for 4 weeks. Then, fatigue, sleep, autonomic nerves, stress, and immunity were evaluated. In addition, overdose safety study of the extract powder (up to 16 mg/day of 6-MSITC for 4 weeks) was performed with 30 healthy volunteers in double-blind placebo-controlled method. Results: Wasabi extract powder containing 6-MSITC did not improve fatigue after mental task, but fatigue before mental task, sleep and mood were improved significantly after 4 weeks intake. No changes were observed in the autonomic nerve function, stress, or immune markers. In the overdose safety study, no changes in the parameters or side effects were observed, and the results showed that overdosing on wasabi extract powder containing 6-MSITC was safe. Conclusion: This study confirmed the possibility that wasabi extract powder containing 6-MSITC may improve brain fatigue and sleep. However, since the effectiveness evaluation in this study was a single-arm, open-label study and there was no placebo control group, these points must be considered when interpreting the results. Safety was confirmed in an overdose study of more than three times the amount compared to that in the efficacy evaluation study. In the future, further research should be conducted on brain fatigue and sleep with wasabi extract powder containing 6-MSITC. Trial registration: UMIN clinical trial registration system, UMIN000049913. Registered 27 December 2022 Retrospectively registered, https://center6.umin.ac.jp/cgiopen-bin/ctr/ctr_view.cgi?recptno=R000056818

In the brain, many regions work in a network-like association, yet it is not known how durable these associations are in terms of activity and could survive without structural connections. To assess the association or similarity between brain regions with a new “generating” approach, this study evaluated the similarity of activities of neurons at the cellular level within each region after disconnecting between regions. To this end, a multi-layer LSTM (Long-Short Term Memory) model was used. Surprisingly, the results revealed that generation of activity from one region to other regions that had been disconnected was possible with similar reproduction accuracy as generation between the same regions in many cases. Notably, not only firing rates but also synchronization of firing between neuron pairs, which is often used as neuronal representations, could be reproduced with considerable precision. Additionally, their accuracies were associated with the relative distance between brain regions and the strength of the structural connections that initially connected them. This outcome not only enables us to look into principles in neuroscience based on the potential to generate new informative data, but also creates neural activity that has not been measured in adequate amounts and could potentially lead to reduced animal experiments.

The brain is an organ that functions as a network of many elements connected in a non-uniform manner. Especially, the cortex is evolutionarily newest, and is thought to be primarily responsible for the high intelligence of mammals. In the mature mammalian brain, all cortical regions are expected to have some degree of homology, but have some variations of local circuits to achieve specific functions enrolled by individual regions. However, few cellular-level studies have examined how the networks within different cortical regions differ. This study aimed to find rules for systematic changes of connectivity (microconnectomes) across 16 different cortical region groups. We also observed unknown trends in basic parameters in vitro such as firing rate and layer thickness across brain regions. The results revealed that the frontal group shows unique characteristics such as dense active neurons, thick cortex and strong connections with deeper layers. This suggests the frontal side of the cortex is inherently capable of driving, even in isolation.This may suggest that deep layers of frontal node provide the driving force generating a global pattern of spontaneous synchronous activity, such as the Default Mode Network. This finding may explain why disruption in this region causes a large impact on mental health.

Background Wasabi (Eutrema japonicum) is a common pungent spice used in Japan. 6-Methylsulfinylhexyl isothiocyanate (6-MSITC) found in the rhizome of wasabi has been shown to have anti-inflammatory and antioxidant effects, as well as improve neuroinflammation and memory. Therefore, we hypothesized that these effects would be beneficial for treating myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). The present study was conducted to investigate the effectiveness of wasabi extract containing 6-MSITC on ME/CFS in an open-label trial. Methods Fifteen patients (3 males, 12 females, 20–58 years old) were orally administered wasabi extract (9.6 mg of 6-MSITC/day) for 12 weeks. The following parameters and test results were compared pre- and post-treatment: performance status (PS), self-rating questionnaires, pressure pain threshold (PPT) on the occiput, Trail Making test-A (TMT-A), and hemodynamic patterns determined by an active standing test. Results After treatment with 6-MSITC, PS improved significantly (p = 0.001). Although the scores on the 11-item Chalder Fatigue scale (CFS-11) and numerical rating scale (NRS) of fatigue did not show significant changes, subjective symptoms improved significantly, including headache frequency (4.1 to 3.0 times/week, p = 0.001) and myalgia (4.1 to 2.4 times/week, p = 0.019), NRS brain fog scores (5.7 to 4.5, p = 0.011), difficulty finding appropriate words (4.8 to 3.7, p = 0.015), photophobia (4.8 to 3.5, p = 0.008), and the Profile of Mood Status vigor score (46.9 to 50.0, p = 0.045). The PPT of the right occiput (17.3 to 21.3 kPa, p = 0.01) and TMT-A scores (53.0 to 38.1 s, p = 0.007) also changed, suggesting reduced pain sensitivity, and improved cognitive function, respectively. Orthostatic patterns determined by a standing test did not show remarkable changes. There were no serious adverse reactions. Conclusion This study suggests that 6-MSITC improves PS as well as subjective symptoms such as pain and cognitive dysfunction, and psychological vitality of patients with ME/CFS. It also improved cognitive performance and increased pain thresholds in these patients. 6-MSITC may be a promising therapeutic option especially for improving cognitive dysfunction associated with ME/CFS.

Chitin is a key component of hard parts in many organisms, but the biosynthesis of the two distinctive chitin allomorphs, α- and β-chitin, is not well-understood. The accurate determination of chitin allomorphs in natural biomaterials is vital. Many chitin-secreting living organisms, however, produce poorly crystalline chitin which leads to spectrums with only broad lines and imprecise peak positions under conventional analytical methods such as X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), and solid state nuclear magnetic resonance spectroscopy (NMR), resulting in inconclusive identification of chitin allomorphs. Here, we developed a novel method for discerning chitin allomorphs based on their different complexation capacity and guest selectivity, using ethylenediamine (EDA) as a complexing agent. From the peak shift observed in XRD profiles of the chitin/EDA complex, the chitin allomorphs can be clearly discerned. By testing this method on a series of samples with different chitin allomorphs and crystallinity, we show that the sensitivity is sufficiently high to detect the chitin allomorphs even in near-amorphous, very poorly crystalline samples. This is a powerful tool for determining the chitin allomorphs in phylogenetically important chitin-producing organisms and will pave the way to clarify the evolution and mechanism of chitin biosynthesis.

Community-based diversity analyses, such as metabarcoding, are increasingly popular in the field of metazoan zooplankton community ecology. However, some of the methodological uncertainties remain, such as the potential inflation of diversity estimates resulting from contamination by pseudogene sequences. Furthermore, primer affinity to specific taxonomic groups might skew community composition and structure during PCR. In this study, we estimated OTU (operational taxonomic unit) richness, Shannon’s H’, and the phylum-level community composition of samples from a coastal zooplankton community using four approaches: complement DNA (cDNA) and genomic DNA (gDNA) mitochondrial COI (Cytochrome oxidase subunit I) gene amplicon, metatranscriptome sequencing, and morphological identification. Results of mismatch distribution demonstrated that 90% is good threshold percentage to differentiate intra- and inter-species. Moderate level of correlations appeared upon comparing the species/OTU richness estimated from the different methods. Results strongly indicated that diversity inflation occurred in the samples amplified from gDNA because of mitochondrial pseudogene contamination (overall, gDNA produced two times more richness compared with cDNA amplicons). The unique community compositions observed in the PCR-based methods indicated that taxonomic amplification bias had occurred during the PCR. Therefore, it is recommended that PCR-free approaches be used whenever resolving community structure represents an essential aspect of the analysis.

Hyperspectral data in the near infrared range were examined for nine common types of plastic particles of 1 mm and 100-500 μm sizes on dry and wet glass fiber filters. Weaker peak intensities were detected for small particles compared to large particles, and the reflectances were weaker at longer wavelengths when the particles were measured on a wet filter. These phenomena are explainable due to the effect of the correlation between the particle size and the absorption of infrared light by water. We constructed robust classification models that are capable of classifying polymer types, regardless of particle size or filter conditions (wet vs. dry), based on hyperspectral data for small particles measured on wet filters. Using the models, we also successfully classified the polymer type of polystyrene beads covered with microalgae, which simulates the natural conditions of microplastics in the ocean. This study suggests that hyperspectral imaging techniques with appropriate classification models allow the identification of microplastics without the time- and labor-consuming procedures of drying samples and removing biofilms, thus enabling more rapid analyses.

The recent influx of microplastics into the Arctic Ocean may increase environmental stress on the western Arctic marine ecosystem, which is experiencing significant sea-ice loss due to global warming. Quantitative data on microplastics in the western Arctic Ocean are very limited, and the microplastic budget of the water column is completely unknown. To fill in gaps in our knowledge of Arctic microplastics, we observed surface concentrations (number of particles per unit volume of seawater) of meso- and microplastics using a neuston net, and we observed wind speeds and significant wave heights in the Chukchi Sea, Bering Strait, and Bering Sea. From these observations, we estimated the total number (particle inventory) and mass (mass inventory) of microplastics in the entire water column by taking into account the effect of vertical mixing. The particle inventory of microplastics in the Chukchi Sea ranged from 0 to 18,815 pieces km

Few studies have investigated the prevalence of polybrominated diphenyl ethers (PBDEs) in deep-sea sharks. In this study, the levels and profiles of PBDEs were determined in liver samples of eight different species of deep-sea sharks collected in Suruga Bay, Japan. Widespread contamination of PBDEs in the deep-sea environment was reconfirmed in this study as these persistent organic pollutants (POPs) were detected in all specimens analyzed. Mean ΣPBDE levels in the deep-sea sharks ranged from 7 to 517 ng/g of lipid weight. The distribution patterns of BDE homologues were similar in all species where tetra-BDEs provided the dominant contribution to total PBDEs (46%). PBDEs levels were similar to, or higher than, those seen in other deep-sea sharks from different regions. The levels of PBDEs were likely to reflect their feeding preferences as higher PBDE levels were seen in species with higher trophic positions.

Graphical abstract, Removing non-plastic materials is a mandatory process for studying microplastics in environmental samples, and non-plastic materials, both inorganic and organic matter, are often removed chemically through sequential processes. In the multiple chemical treatment processes, the samples need to be collected and the reagent removed at the end of each chemical treatment before the samples are again exposed to a different reagent in a separate container. This leads to a loss of microplastics to some extent. Here, we developed a new, yet simple, small sieve made of stainless-steel that can fit in a laboratory beaker (e.g. 200 ml volume), allowing it to be transferred as-is between chemical treatments of environmental samples, even being soakable in a beaker of acid solution. The collection rates of microplastics were significantly higher in the small stainless-steel sieve than the commonly used filter method for different size of microplastic particles. The use of the new sieve means the processes of rinsing off and filtering samples can be abbreviated throughout the entire process of non-plastic matter removal from environmental samples, contributing to a lower chance of microplastic loss. The time consumed in the sieve method was also significantly lower than for the filtering method due to the elimination of the collection and rinsing steps, thus the use of this sieve can reduce processing time for the samples. The new method is innovative in terms of reducing both the microplastic loss and processing time during chemical treatment processes. • The method developed allows the lower chance of microplastic loss during chemical digestion process • The method reduces the time of sequential processes during chemical digestion

Graphical abstract, Microplastics are abundant even on the deep-sea floor far from land and the ocean surface where human activities take place. To obtain samples of microplastics from the deep-sea floor, a research vessel and suitable sampling equipment, such as a multiple corer, a box corer, or a push corer manipulated by a remotely operated (ROV) or human occupied vehicle (HOV) are needed. Most such corers use sampling tubes made of plastic, such as polycarbonate, acrylic, or polyvinyl chloride. These plastic tubes are easily scratched by sediment particles, in particular during collection of coarse sandy sediments, and, consequently, the samples may become contaminated with plastic from the tube. Here, we report on the use of aluminum tubes with both a multiple corer and a push corer to prevent such plastic contamination. When compared with plastic tubes, aluminum tubes have the disadvantages of heavier weight and non-transparency. We suggest ways to overcome these problems, and we also present an onboard processing protocol to prevent plastic contamination during sediment core sampling when plastic tubes are used. • Use of a sediment corer with aluminum tubes reduces the risk of plastic contamination in the sediment samples • The proposed method allows undisturbed sediment cores to be retrieved with comparable efficiency to conventional transparent core tubes

The deep-sea is considered to be an ultimate sink for marine plastic debris. The abyssal (3500-6500 m) covers the bulk of the deep ocean floor yet little is known about the extent of plastic debris on the abyssal seafloor. We undertook a quantitative assessment of debris presents on the abyssal seafloor (5700-5800 m depth) beneath the Kuroshio Extension current system in the Northwest Pacific, which is one of the major transit points for massive amounts of debris sourced from Asia that are entering the North Pacific Ocean. The dominant type of debris was single-use plastics-mainly bags and food packaging. The density of plastic debris (average 4561 items km−2) in the abyssal zone was the highest recorded for an abyssal plain, suggesting that the deep-sea basin of the Northwest Pacific is a significant reservoir of plastic debris.

The retinoic acid receptor-related orphan nuclear receptor gamma t (RORγt), which is a promising therapeutic target for immune diseases, is a major transcription factor of genes related to psoriasis pathogenesis, such as interleukin (IL)-17A, IL-22, and IL-23R. Inspired by the co-crystal structure of RORγt, a 6-oxo-4-phenyl-hexanoic acid derivative 6a was designed, synthesized, and identified as a ligand of RORγt. The structure-activity relationship (SAR) studies in 6a, which focus on the improvement of its membrane permeability profile by introducing chlorine atoms, led to finding 12a, which has a potent RORγt inhibitory activity and a favorable pharmacokinetic profile.

Plastic waste has become a growing concern in terms of marine pollution, but little information is available on plastic debris and its possible risks of chemical additives exposure in the deep-sea. This study focused on identification of polymer type and additive concentrations in 21 plastic debris collected from deep-sea of Sagami Bay, Japan and West Pacific Ocean under the Kuroshio Extension and its recirculation gyre (KERG) zone (water depth: 1388–5819 m). Polyethylene (PE) was dominant polymer (57% of the total) in samples, followed by polyvinylchloride (PVC), epoxy resin, polyester (PES), and polypropylene. In plastic additives, bis (2-ethylhexyl) phthalate (DEHP) was detected to be contained in a PVC sheet at concentration of 48%. Butylated hydroxytoluene (BHT) was also detected in PE plastic debris with median concentration of 12,000 ng/g. PES clothes were detected to contain dyeing mixtures, 1,2,4-trichlorobenzene (1,2,4-TCB), up to 42,000 ng/g. Knowing the estimated number of plastic debris under KE current, the minimum burden of chemical additives were estimated that 720 kg of dibutyl phthalate, 570 kg of BHT, 230 kg of DEHP, and 160 kg of 1,2,4-TCB exist on the seabed of KERG zone. This result strongly suggests that enormous amount of hazardous additives lie within plastic debris on abyssal level of the ocean.

Plastic pollution has become one of the most emergent issues threating aquatic and terrestrial ecosystems. However, it is still challenging to rapidly detect small microplastics. Here, we present a method to rapidly detect microplastics using hyperspectral imaging in which we optimized a commercially available hyperspectral imaging system (Pika NIR-640, Resonon Inc., USA). The optimizations included: (1) changing the four-lamp assembly to a symmetrical set of converged-light near-infrared lamps that are placed sideways instead of above the sample stage; (2) adopting a macro-photography technique by applying an extension tube between the camera and the lens, and moving the lens of the hyperspectral camera to the imaging target (working distance of ~3 cm); (3) adjusting the exposure and aspect ratio by tuning the frame rate and scan speed of the imaging system. After optimization, the detection resolution of each pixel improved from 250 µm to 14.8 µm. With the optimized system, microplastics down to 100 µm in size were rapidly detected. This result is promising for the application of our new method in the accelerated detection of microplastics and will contribute to a better understanding of the microplastic pollution situation., Graphical abstract Image, graphical abstract

The relative contributions of grazing versus microbial food webs to the production of mesozooplankton communities in coral reef ecosystems remains an important and understudied field of inquiry. Here, we investigated the biomass and production of component organisms within these two food webs, and compared them to those of mesozooplankton on a coral reef in Okinawa, Japan throughout four seasons in 2011–2012. The relative production of grazing (phytoplankton) and microbial (nano and microzooplankton) food webs were on average 39% (7–77%) and 37% (19–57%), respectively, of the food requirements of particle-feeding mesozooplankton. Carbon flows within this planktonic food web suggested that primary production from the grazing food web could not satisfy the nutritional demands of mesozooplankton, and that the microbial food web contributed a significant amount of nutrition to their diets. These results also show that the heterotrophic components of the microbial food web (nano and microzooplankton) and mesozooplankton consume the equivalent of the entire phytoplankton production (particulate net production) each day, while the microzooplankton were almost entirely eaten by higher trophic levels (mesozooplankton) each day. However, even the combined production from both the grazing and microbial food webs did not fulfill mesozooplankton food requirements in some seasons, explaining 26–53%, suggesting that detritus was used to compensate for nutritional deficiencies during these periods. Understanding the flow of energy throughout coral reefs requires a detailed accounting of pelagic sources and sinks of carbon. Our results provide such an assessment and indicate that detailed investigation on the origin and production of detritus is necessary to better understand pelagic trophodynamics in coral ecosystems.

Exudates derived from hermatypic corals were incubated with

Daily field surveys were conducted at a coastal-shelf station in Sagami Bay, Japan after the passage of typhoon Malou in 2010 to evaluate the after-effect of a typhoon passage on the physical-chemical environment, phytoplankton bloom formation and microbial processes within and below the euphotic layer. The passage of Malou induced an abrupt decrease in salinity and increased loading of nutrients to the euphotic layer. Dinoflagellates dominated the phytoplankton community at the surface, whereas diatoms dominated below the surface just after the passage of Malou. Four days later, the dominant dinoflagellate taxa at the surface changed from Protoperidinium spp. to Prorocentrum spp. and Ceratium spp., indicating a dinoflagellate community succession from heterotrophic to autotrophic functional groups. Five days after passage, the dominant phytoplankton taxa shifted from dinoflagellates to diatom groups of Chaetoceros spp. and Cerataulina spp. throughout the water column. Below the euphotic layer, there were increases in diatom frustules, mainly composed of Chaetoceros spp. and Cerataulina spp., bacterial abundance and NH4+ concentrations. Diatom carbon biomass contributed to approximately half of particulate organic carbon (POC) below the euphotic layer, suggesting a significant contribution of diatoms to POC sinking flux after the passage of a typhoon. Bacterial abundance was positively correlated to both phaeopigment concentrations (p

Microplastic pollution has become an urgent issue because it adversely affects ecosystems. However, efficient methods to detect and characterize microplastic particles are still in development. By conducting a series of laboratory assessments based on near-infrared hyperspectral imaging in the wavelength range of 900–1700 nm, we report the fundamental spectral features of (i) 11 authentic plastics and (ii) 11 filter substrate materials. We found that different plastic polymers showed distinct spectral features at 1150–1250 nm, 1350–1450 nm and 1600–1700 nm, enabling their automatic recognition and identification with spectral separation algorithms. Using an improved hyperspectral imaging system, we demonstrated the detection of three types of microplastic particles, polyethylene, polypropylene and polystyrene, down to 100 μm in diameter. As a filter substrate, a gold-coated polycarbonate filter (GPC0847-BA) showed constant reflectance over 900–1700 nm and a large radiative contrast against loaded plastic particles. Glass fiber filters (GF10 and GF/F) would also be suitable substrates due to their low cost and easy commercial availability. This study provides key parameters for applying hyperspectral imaging techniques for the detection of microplastics.

Cabled seafloor platforms can overcome many constraints of stand-alone systems and archived long-term seafloor monitoring, and their advantages include unlimited power, real-time communication, a synchronized clock, and virtually unlimited data storage. Using monthly images from a fixed camera on a cabled deep-sea observatory in the Off Hatsushima seep site in Sagami Bay, Japan, fluctuations in clam density (Phreagena soyoae and P. okutanii) were observed over 5 years, March 1994–July 1999. Eight disturbances were seen, including five phytodetritus deposition events associated with seasonal blooms, two earthquake-associated turbid flows, and one caused by a maintenance visit by a remotely operated vehicle. Overall, these observations indicate that the vesicomyids experienced an ongoing decline throughout the observation series, and strong disturbance events such as earthquakes appeared to be too severe for the clams to survive. As such, vesicomyid clams are unlikely to survive the foreseeable impacts of deep-sea mining. These results show that long-term observations of deep-sea chemosynthetic communities are useful for understanding their resilience and for making an environmental impact assessment.

Most cells for regenerative medicine are currently cultured manually. In order to promote the widespread use of regenerative medicine, it will be necessary to develop automated culture techniques so that cells can be produced in greater quantities at lower cost and with more stable quality. In the field of regenerative medicine technology, cell sheet therapy is an effective tissue engineering technique whereby cells can be grafted by attaching them to a target site. We have developed automated cell culture equipment to promote the use of this cell sheet regenerative treatment. This equipment features a fully closed culture vessel and circuit system that avoids contamination with bacteria and the like from the external environment, and it was designed to allow 10 cell sheets to be simultaneously cultured in parallel. We used this equipment to fabricate 50 sheets of human oral mucosal epithelial cells in five automated culture tests in this trial. By analyzing these sheets, we confirmed that 49 of the 50 sheets satisfied the quality standards of clinical research. To compare the characteristics of automatically fabricated cell sheets with those of manually fabricated cell sheets, we performed histological analyses using immunostaining and transmission electron microscopy. The results confirmed that cell sheets fabricated with the automated cell culture are differentiated in the same way as cultures fabricated manually.

After the passage of typhoon Malou in 2010, daily field samplings were conducted at both inshore (Sta. A) and offshore (Sta. M) stations in Sagami Bay, Japan, to evaluate responses of bacteria and phytoplankton to variations of physical-chemical environments induced by typhoon passage. Malou passage caused an abrupt decline of salinity and a large increase in the amount of nutrients at both stations. The relationships between salinity and nutrient concentrations suggested that major nutrient sources were terrestrial runoff at Sta. M and sediment resuspension in addition to terrestrial runoff at Sta. A. Bacterial production (BP) at Sta. A showed 114 ± 21 mg C m −3 d −1 one day after Malou passage, while primary production (PP) was 76 ± 8 mg C m −3 d −1 , suggesting the dominance of BP (BP/PP ratio = 1.5). PP exceeded BP two days after Malou passage, and then reached a maximum of 554 ± 32 mg C m −3 d −1 five days later (BP/PP ratio = 0.10). PP was always dominant at Sta. M throughout the study period (BP/PP ratio = 0.13 ± 0.05). The ratio of BP to bacterial abundance (BP/BA ratio) at Sta. M showed a positive correlation with PP, suggesting that bacterial productivity depended on autochthonous substrates derived from phytoplankton. The BP/BA ratio at Sta. A showed no relationship with PP, suggesting that bacterial productivity was enhanced not only by PP, but also loading of allochthonous substrates. BP/BA ratios at both stations increased exponentially with the increase of PO 4 and NH 4 concentrations; these concentrations are likely coming from sediment pore waters. The results suggest that sediment resuspension induced by typhoon passage enhanced bacterial productivity abruptly just after the passage at an inshore station. The bacterial response could be regulated by difference in relative contribution of nutrient sources after the passage of typhoon.

Egg surface ornamentation, egg production rates, and hatching success of two calanoid copepod species (Acartia pacifica and Acartia cf. erythraea) were examined in a tropical coral reef of Tioman Island, Malaysia, near the equator (2°N). All observed females of A. pacifica and A. cf. erythraea produced eggs with spines on the surface, either short or long, which are common in resting eggs in temperate ecosystems that hatch when environmental conditions are favorable or after a certain refractory period. However, all of these spiny eggs in the present study hatched within 48 hours when exposed to ambient seawater temperature, suggesting that they were subitaneous eggs. Such spine ornamentation has been thought to be an adaptive feature only for resting eggs to survive during egg dormancy, but the presence of spines on subitaneous eggs suggests further possible functions of these spines such as reducing the risks of ingestion by predators.

Dissolved organic matter (DOM) constitutes the largest organic matter pool in coral reef waters and is released and utilized by various coral reef organisms. In this chapter, we review the distribution and fluctuation of DOM concentrations in coral reefs around the world, with a special focus on Shiraho Reef, Ishigaki Island, Okinawa, Japan, where DOM fluxes have been studied most intensively since the late 1990s. Then, we review the DOM production rates from specific reef organisms and DOM consumption rates by bacteria. Previous studies have shown that both dissolved organic carbon and nitrogen (DOC and DON, respectively) generally have a higher concentration in most coral reefs than in the surrounding ocean. At Shiraho Reef, the average ratio of the net DOC production to the net primary production on the reef flat was 18%, and the C:N ratio of DOM that was produced on the reef flat was estimated to be 9.3. The abundance of heterotrophic bacteria was also higher in most coral reefs than offshore, which indicates that bacterial growth was enhanced by reef-derived DOM. Some of the DOC that was produced in coral reefs was persistent to bacterial decomposition in the long term, which suggests that coral reef ecosystems export some reef-derived DOM to the ambient ocean, irrespective of the water residence time in the reef.

In order to test the hypothesis that species composition of planktonic copepods varies with seasonal monsoons in tropical coral reef waters of South East Asia, we investigated the copepod community structure in a coral reef of Malaysia over different monsoonal periods (the southwest, the northeast and inter-monsoons). Small copepods such as Paracalanus elegans and Oithona spp. were the predominant taxa, comprising 65%–78% of the total copepod abundance and occur throughout the year. The relative abundance of these dominant copepod species showed no clear seasonal pattern in abundance, suggesting that their populations are relatively stable. Yet, significant differences in the composition and abundance of the remaining non-dominant species resulted in distinctive seasonal communities of copepods being detected. Multivariate multiple regressions revealed that the community structure and species composition fluctuates with monsoonal variables (wind velocity and direction, water temperature) and POC concentration, emphasizing that the differences in water source and food availability are important factors in shaping the distinct structure of the copepod assemblages in tropical coral reef waters of South East Asia.

In order to examine small invertebrate scavenging fauna of tropical coral reef waters, baited traps were deployed in a coral reef of Bidong Island, Malaysia. The samples taken by the traps constituted only a single species of isopods (Cirolana sp.) and ostracods from three families (Cypridinidae, Cylindroleberididae, and Paradoxostomatidae). Cirolanid isopods were never collected in net-samples suggesting they are hyperbenthic scavenging species. Judging from its sheer numbers in the bait-attracted community (86.1-98.6%), cirolanid isopods are one of the most important small invertebrate scavengers in coral reef waters of Bidong Island. Abstrak Di dalam mengetahui kehadiran fauna invertebrat pemangsa di kawasan terumbu karang di perairan tropika beberapa perangkap berumpan telah diletakkan di sekitar kawasan terumbu karang di Pulau Bidong. Perangkap berumpan tersebut telah berjaya memerangkap hanya satu spesies tunggal isopod (Cirolana sp) dan juga tiga famili ostrakod (Cypridinidae, Cylindroleberididae, dan Paradoxostomatidae). Didapati, isopod cirolanid tidak pernah direkodkan kehadirannya di dalam mana-mana persampelan yang menggunakan net, menunjukkan bahawa Malaysian Journal of Science 32 (2): 59-66 (2013) 60 kumpulan organisma ini adalah sebagai spesies pemangsa hiperbentik. Merujuk kepada kehadirannya yg amat tinggi di dalam komuniti perangkap-berumpan tersebut (86.1-98.6%), didapati isopod cirolanid ini merupakan salah satu kumpulan invertebrate pemangsa kecil yang penting di kawasan terumbu karang di Pulau Bidong. (Keywords: Bait-trap, Cirolana isopods, Coral reef, Ostracods, Scavenger)

A sophisticated method was developed for evaluating simultaneously and accurately both the average specific resistance and average porosity of the filter cake formed in unstirred dead-end ultrafiltration of nanocolloids such as protein solution and nanosilica sol. In the method, a step-up pressure filtration test was conducted by using a filter with a single-stage reduction in the effective filtration area. The influence of the pressure drop across the cake on not only the average specific cake resistance but also on the average cake porosity of highly compressible filter cake was evaluated using only flux decline data in one dead-end filtration test, taking advantage of the decrease in the cake thickness caused by the pressure increase. As a result, the cake properties were easily determined for a variety of nanocolloids. Constant pressure dead-end ultrafiltration data obtained under various pressures and concentrations were well evaluated based on the method proposed. © 2015 American Institute of Chemical Engineers AIChE J, 61: 4426–4436, 2015

After the adoption of the Aichi Target, data accumulation and evaluation regarding biodiversity have progressed rapidly. The use of ecologically and biologically significant areas (EBSAs) criteria to evaluate important areas enables the identification of effective and prioritized areas for ecosystem management. This includes strategic environmental assessment and discussions aimed at establishing protected marine areas based on scientific data. This paper reviews previous and current ideas as well as the methods used, for the identification of EBSAs. In particular, the following issues are addressed: problems associated with different types of marine ecosystems in the Japanese Archipelago, such as seagrass and seaweed beds, coral reefs, offshore pelagic plankton, and deep-sea benthic ecosystems; and problems associated with the integration of multiple criteria that are not totally exclusive. Several candidate variables accounting for each of the 7 criteria used to identify ecologically important areas are presented. Data availability is the most important criterion that allowed for the comprehensive evaluation of different types of ecosystems in the same localities. In particular, for coastal ecosystems such as seagrass, seaweed beds, and coral reefs, it is possible to carry out broad spatial comparisons using variables representing most of these 7 criteria. Regarding methods for the quantitative evaluation of each criterion and their integration, application of these methods to kelp forest ecosystems in Hokkaido, Northern Japan is presented as a case study.

Summary Background Epidermal cell sheets have been utilized for regeneration of skin when skin defects occur and prevention of esophageal stricture after endoscopic submucosal dissection. To reduce the cost of cultivation, a novel culture method to shorten a culture process needs to be developed. Objectives To shorten a culture process of epidermal cell sheets, we developed a novel culture method to accelerate the fabrication of epidermal cell sheets using γ-secretase inhibitor. Methods Normal human epidermal keratinocytes (NHEKs) were cultured using γ-secretase inhibitor, DAPT, during expansion of the cells to confluence and culture without DAPT during stratification. The cell growth, quantitative gene expression of stem/progenitor or differentiation markers, and protein expression of these markers were analyzed to verify the effectiveness of the novel method. Results The proliferation of NHEKs on cell-culture inserts was promoted using DAPT. However, NHEKs were not stratified completely in the presence of DAPT. In contrast, NHEKs cultured using DAPT were stratified and differentiated by eliminating the inhibitor after the cells reached confluence. Real-time PCR analyses showed that the gene expressions of putative epithelial stem/progenitor cell markers and epidermis differentiation markers in the cell sheets fabricated using this novel method were significantly higher than those in the cell sheets fabricated without DAPT. Histological and immunofluorescence analyses revealed that it was possible to fabricate well-differentiated epidermal cell sheets efficiently by the novel culture method. The culture period was shortened to 67% of the time required for the control group. In feeder-free conditions, stratified epidermal cell sheets were also fabricated using DAPT. Conclusions The novel culture method using γ-secretase inhibitor, DAPT, was found to be effective for fabricating epidermal cell sheets.

A potential method has been developed for evaluating simultaneously both the average specific resistance and average porosity of the filter cake formed in unstirred dead-end ultrafiltration of nanocolloids such as bovine serum albumin solution and silica sol. The method consists of variable pressure filtration followed by constant pressure filtration. The relation between the average specific cake resistance and the pressure drop across the cake was determined from the evolution of the filtration rate with time in the course of the variable pressure filtration period, based on the compressible cake filtration model. The average porosity was evaluated from the significant flux decline caused by a sudden reduction in the cake surface area in the middle of the constant pressure filtration period. The pressure dependences of both the average specific cake resistance and average cake porosity were obtained from only two runs which differed from each other in the pressure profiles. © 2014 American Institute of Chemical Engineers AIChE J, 60: 3869–3877, 2014

An ITO (indium-tin-oxide)-nanoparticle film electrode was prepared by applying an ITO nanoparticle-dispersed solution to an ITO planar plate and then sintering it. The electrochromic properties of 1,1′-dibenzyl-4,4′-bipyridinium (DBV2+) were examined at the nanoparticle film electrode. The one-electron reduction of DBV2+ produced a filmy purple deposit of DBV+, and electrochromism between the white and purple appeared. An incomplete vanishing (ghost image), which often occurs when an ITO planar electrode is used, was not observed at the nanoparticle film electrode. The successful ghost-free electrochromism was explained in terms of the much lower molecular density of the deposited DBV+ per unit real surface area compared to that on the ITO planar electrode. Also, it was demonstrated that the coloration–decoloration cycle was significantly improved by the effect of the ghost-free electrochromism.

Inorganic nutrient contents of mucus released by Acropora corals and its utili-zation by heterotrophic bacteria at several different hour intervals in the coralmucus were investigated at a coral reef in Malaysia. The dissolved inorganicphosphate (DIP) concentration was 135-fold higher than in the ambient seawa-ter, probably due to inorganic P release from the coral gut cavity. We experi-mentally confirmed that heterotrophic bacteria rapidly (within 8 h) consumedca. 80% of the initial concentration of DIP derived from coral mucus. HighDIP concentration in coral mucus may enhance heterotrophic bacterial pro-duction and associated carbon flow in the microbial loop of reef ecosystems.IntroductionThe photosynthetically fixed carbon translocated to thecoral host from endosymbiotic zooxanthellae represents aprimary source of energy for scleractinian corals (e.g.Lesser et al. 2000). However, corals exude up to half ofthe organic carbon provided by zooxanthellae into thesurrounding water (e.g. Crossland et al. 1980) for feeding,shedding of sediments, defense against pathogens, protec-tion against UV radiation, and desiccation resistance(reviewed by Wild et al. 2004a). This organic matter isoften referred to as coral mucus. Previous studies havefound that ca. 80–90% of coral mucus is labile and thusfunctions as a suitable growth substrate for heterotrophicbacteria (hereafter bacteria) in the seawater (Tanaka et al.2011), enabling high bacterial production in coral reefecosystems (e.g. Ferrier-Pages et al. 2000).Live corals continuously release mucus into the ambi-ent water (Naumann et al. 2010) and are apparently themain source for organic carbon in coral reef waters (vanDuyl & Gast 2001). In such an environment, inorganicnutrients could potentially be a limiting factor for bacte-rial growth (van Duyl & Gast 2001) because bacterialproduction is stimulated further when inorganic nutrientsare added to labile carbon (e.g. Zweifel et al. 1993; Cotneret al. 1997; Carlson et al. 2002; Sharon & Rosenberg2008). Bacteria acquire a significant fraction of their Nand P demand via uptake of inorganic forms of these ele-ments (Kirchman 2000), although the availability of Nand P in labile dissolved organic matter can influenceinorganic uptake demand. In oligotrophic reef waters(Sheppard et al. 2009), coral mucus may be an importantinorganic nutrient source for bacterial growth in coralreefs.At present, little is known about the inorganic nutrientcontents of coral mucus (Wild et al. 2005; Nakajimaet al. 2009), although organic carbon of coral mucus byvarious corals has been measured in several studies (e.g.

Aim To characterize the species richness, endemism and community structure of benthic macro-/megafauna in the deep-sea chemosynthetic ecosystems around the Japanese archipelago and determine the conservation priority for deep-sea chemosynthetic habitats. Location Twenty-seven methane seeps from the Kuril–Japan Trench, Nankai Trough, Ryukyu Trench and Sagami Bay as well as 15 hydrothermal vents from the Izu–Bonin Arc and Okinawa Trough around the Japanese archipelago. Methods Species richness and the number of endemic species from 42 vents and seeps were examined using the distributional data of 155 mollusc, annelid and arthropod species. Multivariate analyses were performed to identify groups of sites with similar assemblage composition. Complementarity analysis was performed to determine the minimum number of sites required to conserve at least one population of all species for all 42 sites and for groups of sites detected by the multivariate analyses. Results The species richness of the deep-sea chemosynthetic sites around the Japanese archipelago was higher in the Okinawa Trough and Sagami Bay where higher methane concentrations were observed. Eight distinct biogeographic regions were detected at the community level, and the community structure was classified according to the geological variables of depth and latitude. Complementarity analysis for all 42 sites indicated 21 sites for conservation, while that for each distinct biogeographic region indicated 30 sites for conservation. Main conclusions As unique communities exist in each geographically isolated region around the Japanese archipelago, priority areas should be identified for each type of bioregion for effective conservation. At least 30 (17 seeps and 13 vents) of 42 sites should be conserved to avoid the extinction of the local populations of deep-sea chemosynthetic macro-/megabenthos.

The use of murine 3T3 feeder cells needs to be avoided when fabricating corneal epithelial cell sheets for use in treating ocular surface diseases. However, the expression level of the epithelial stem/progenitor cell marker, p63, is down-regulated in feeder-free culture systems. In this study, in order to fabricate corneal epithelial cell sheets that maintain colony-forming cells without using any feeder cells, we investigated the use of an oxygen-controlled method that was developed previously to fabricate cell sheets efficiently. Rabbit limbal epithelial cells were cultured under hypoxia (1–10% O 2 ) and under normoxia during stratification after reaching confluence. Multilayered corneal epithelial cell sheets were fabricated using an oxygen-controlled method, and immunofluorescence analysis showed that cytokeratin 3 and p63 was expressed in appropriate localization in the cell sheets. The colony-forming efficiency of the cell sheets fabricated by the oxygen-controlled method without feeder cells was significantly higher than that of cell sheets fabricated under 20% O 2 without feeder cells. These results indicate that the oxygen-controlled method has the potential to achieve a feeder-free culture system for fabricating corneal epithelial cell sheets for corneal regeneration.

In order to extract quantitative information on deep-sea benthic animals (no. individuals or biomass in an area) using oblique video/photo images taken by deep-sea submersible survey platforms, a new method was established to estimate the imaged area of the seafloor from the oblique images. The trapezoidal area appearing on the lower half of the screen was calculated using underwater horizontal and vertical aperture angles of the camera, the angle of incidence of the camera, and the camera-to-seafloor distance. The incidence angle of the camera was obtained using the angles of vehicle pitch and camera tilt, while the camera-to-seafloor distance was calculated from the lens-to-vehicle bottom distance, horizontal distance of lens-to-altimeter, and vehicle altitude. The areas estimated by the present method from images taken by some submersible survey platforms were comparable to those that were actually measured. With the above parameters, and by focusing on the lower half of an image, our method can be used for estimating the seafloor area from any oblique video/photo images taken by any submersible survey platform. Thus, this method is useful for the extraction of quantitative data on benthic animals from legacy oblique video/photographs acquired by submersible survey platforms.