Asian dust exposure triggers acute myocardial infarction.

AIMS: To elucidate whether Asian dust is associated with the incidence of acute myocardial infarction (AMI) and to clarify whether patients who are highly sensitive to Asian dust will develop AMI. METHODS AND RESULTS: Twenty-one participating institutions located throughout Kumamoto Prefecture and capable of performing coronary intervention were included in the study. Data for ground-level observations of Asian dust events were measured at the Kumamoto Local Meteorological Observatory. Data collected between 1 April 2010 and 31 March 2015 were analysed, and 3713 consecutive AMI patients were included. A time-stratified case-crossover design was applied to examine the association between Asian dust exposure and AMI. The occurrence of Asian dust events at 1 day before the onset of AMI was associated with the incidence of AMI [odds ratio (OR), 1.46; 95% confidence interval (CI), 1.09-1.95] and especially, non-ST-segment elevation myocardial infarction was significant (OR 2.03; 95% CI, 1.30-3.15). A significant association between AMI and Asian dust was observed in patients with age ≥75 years, male sex, hypertension, diabetes mellitus, never-smoking status, and chronic kidney disease (CKD). However, Asian dust events had a great impact on AMI onset in patients with CKD (P < 0.01). A scoring system accounting for several AMI risk factors was developed. The occurrence of Asian dust events was found to be significantly associated with AMI incidence among patients with a risk score of 5-6 (OR 2.45; 95% CI: 1.14-5.27). CONCLUSION: Asian dust events may lead to AMI and have a great impact on its onset in patients with CKD.

Lipid droplets of arbuscular mycorrhizal fungi emerge in concert with arbuscule collapse.

Plants share photosynthetically fixed carbon with arbuscular mycorrhizal (AM) fungi to maintain their growth and nutrition. AM fungi are oleogenic fungi that contain numerous lipid droplets in their syncytial mycelia during most of their life cycle. These lipid droplets are probably used for supporting growth of extraradical mycelia and propagation; however, when and where the lipid droplets are produced remains unclear. To address these issues, we investigated the correlation between intracellular colonization stages and the appearance of fungal lipid droplets in roots by a combination of vital staining of fungal structures, selective staining of lipids and live imaging. We discovered that a surge of lipid droplets coincided with the collapse of arbuscular branches, indicating that arbuscule collapse and the emergence of lipid droplets may be associated processes. This phenomenon was observed in the model AM fungus Rhizophagus irregularis and the ancestral member of AM fungi Paraglomus occultum. Because the collapsing arbuscules were metabolically inactive, the emerged lipid droplets are probably derived from preformed lipids but not de novo synthesized. Our observations highlight a novel mode of lipid release by AM fungi.

Isolation and phenotypic characterization of Lotus japonicus mutants specifically defective in arbuscular mycorrhizal formation.

Several symbiotic mutants of legume plants defective in nodulation have also been shown to be mutants related to arbuscular mycorrhizal (AM) symbiosis. The origin of the AM symbiosis can be traced back to the early land plants. It has therefore been postulated that the older system of AM symbiosis was partially incorporated into the newer system of legume-rhizobium symbiosis. To unravel the genetic basis of the establishment of AM symbiosis, we screened about 34,000 plants derived from ethyl methanesulfonate (EMS)-mutagenized Lotus japonicus seeds by microscopic observation. As a result, three lines (ME778, ME966 and ME2329) were isolated as AM-specific mutants that exhibit clear AM-defective phenotypes but form normal effective root nodules with rhizobial infection. In the ME2329 mutant, AM fungi spread their hyphae into the intercellular space of the cortex and formed trunk hyphae in the cortical cells, but the development of fine branches in the arbuscules was arrested. The ME2329 mutant carried a nonsense mutation in the STR-homolog gene, implying that the line may be an str mutant in L. japonicus. On the ME778 and ME966 mutant roots, the entry of AM fungal hyphae was blocked between two adjacent epidermal cells. Occasionally, hyphal colonization accompanied by arbuscules was observed in the two mutants. The genes responsible for the ME778 and ME966 mutants were independently located on chromosome 2. These results suggest that the ME778 and ME966 lines are symbiotic mutants involved in the early stage of AM formation in L. japonicus.

Aerosol soluble proteins in Asian dust in southwestern Japan.

Aerosol proteins, as core biological components of bioaerosols, are garnering increasing attention due to their environmental significance, including their roles in atmospheric processes and associated health risks. However, observational data on the proteins are very limited, leaving their distribution and variation in the atmosphere poorly understood. To investigate the long-distance transport of proteins with Asian dust in the Northern Hemisphere middle latitude westerlies to remote downwind areas, we quantified the soluble proteins in aerosol particles, referred to as aerosol soluble proteins (ASPs), collected in the coastal city of Kumamoto, Japan, during the spring of 2023, when three dust events occurred. The concentration of ASPs ranged from 0.22 to 1.68 µg m-3, with an average concentration of 0.73 ± 0.36 µg m-3 under dust conditions and 0.31 ± 0.05 µg m-3 under non-dust conditions. During the dust periods, the largest concentration of ASPs (1.68 µg m-3) coincided with the peak concentration of suspended particulate matter, and the concentration strongly correlated with the mass concentration of particles larger than 2.5 µm, indicating a close dependence of ASPs on dust particles. Primary estimations indicated a dry deposition flux of ASPs at approximately 1.10 ± 0.87 mg m-2 d-1 under the dust conditions. These results prove that Asian dust efficiently transports proteins, facilitating their dispersion in the atmosphere.

High heterogeneity and aging state of mineral particles in a slowly-moving dust plume on the southwestern coast of Japan.

Aerosol particles in two size ranges, namely 0.18-1.4 µm (fine) and larger than 1.4 µm (coarse), were collected in the pre-dust, in-dust, and post-dust air during the passage of a slowly-moving dust event at a coastal site in southwestern Japan. We identified the composition and size of individual particles using a scanning electron microscope to investigate the variations during dust passage. The particles could be classified as mineral-seasalt mixtures, non-mixture minerals, sulfur-containing minerals, and seasalt particles, and the number fractions of these type particles in the two size ranges exhibited significant variation across the three periods. In the coarse size range, mixture particles accounted for 17.6 %, 26.8 %, and 37.8 % of the particles in the pre-dust, in-dust, and post-dust air, respectively. Non-mixture particles made up 36.8 %, 29.2 %, and 24.3 % in the same respective periods. In the in-dust air, the average relative ratio of sulfur content in sulfur-containing mineral particles in the coarse range was 5.5 %, whereas in the fine range, it was 17.2 %. The aging state of sea salt components, described by the Cl loss and reflecting the changes in particles due to chemical reactions, exhibited significant differences in the two size ranges. In the fine range, the aging of >90 % particles was predominantly influenced by sulfate formation in the in-dust air. In contrast, nitrate likely played a certain role in both the pre-dust and post-dust air. In the coarse range, the aging was independent of sulfate formation. These results indicate the close dependence of the aging of dust particles on their size and the notable variations of the aged states, underscoring the essentiality to treat dust particles properly according to time and space for a better understanding on their roles in the marine atmosphere.

Generic UPLC-MS/MS and Gyrolab assays with blood microsampling for pharmacokinetic assessments of therapeutic antibodies in mice.

Serial blood sampling from one animal is useful to understand relationship between pharmacokinetics (PK) and pharmacological or toxicological events in individual animals. To assess its feasibility in mice, two therapeutic antibodies were used to evaluate impacts by different blood sampling methods, sampling sites, and assay platforms on PK. Denosumab and Panitumumab were intravenously administered to mice and only 0.05 mL of blood sample per point was collected from jugular vein or tail vein. Blood samples were collected serially from a mouse or collected by traditional composite sampling from each mouse. Plasma concentrations of the two drugs were assayed by a generic ligand binding assay using Gyrolab or by a generic ultra-performance liquid chromatography with tandem mass spectrometry. The two assay platforms showed acceptable accuracy and precision and gave comparable PK parameters of the drugs, suggesting that both assays were successfully applied to the PK assessments. Comparable results in the PK profiles were noted between serial and composite blood samplings and differences in the two sampling sites did not impact PK. These findings suggest that microsampling combined with generic assays is useful to assess PK profiles of therapeutic antibodies in mice.

Genomic features of Lactococcus lactis IO-1, a lactic acid bacterium that utilizes xylose and produces high levels of L-lactic acid.

Lactococcus lactis IO-1 (JCM7638) produces L-lactic acid predominantly when grown at high xylose concentrations, and its utilization is highly desired in the green plastics industry. Therefore it is worthwhile studying its genomic traits. In this study, we focused on (i) genes of possible horizontal transfer derivation (prophages, the nisin-sucrose transposon, and several restriction-modification systems), and (ii) genes for the synthetic pathways of amino acids and vitamins in the IO-1 genome. In view of the results of this analysis, we consider their meanings in strain IO-1.

Number size distribution of bacterial aerosols in terrestrial and marine airflows at a coastal site of Japan.

Size-differentiated concentration of bacterial aerosols is essential for investigating their dissemination via the atmosphere. In this study, the number size distribution of bacterial aerosols was measured at a coastal site in southwestern Japan (32.324°N, 129.993°E) using a size-segregated eight-stage (>11, 7.0-11, 4.7-7.0, 3.3-4.7, 2.1-3.3, 1.1-2.1, 0.65-1.1, and 0.43-0.65µm) sampler. The results showed that the distribution differed according to the source areas: terrestrial air, oceanic air, or a combination of the two. The distribution in the long-distance transported terrestrial air from the Asian continent was monomodal, with a peak of 3.3-4.7 µm. The distribution in local land breeze air was bimodal, with the peaks at 0.43-1.1 and 3.3-4.7 µm. A similar bimodal distribution was encountered when the local island air and long-distance transported terrestrial air mixed. In contrast, the size distribution did not show clear peaks in the air from either nearby or remote marine areas. According to the air mass backward trajectories, the further the distance the air moved in the 72 h before arriving at the site, the lower the concentration of total bacterial aerosols. The estimation of dry deposition fluxes of bacterial cells showed that the deposition was dominated by cells larger than 1.1 µm with a relative contribution from 70.5 % to 93.7 %, except for the local land breeze cases, where the contributions in the size ranges larger and smaller than 1.1 µm were similar. These results show the distinctive number size distributions and removal processes of bacterial aerosols in different types of air. In addition, they indicate that size-dependent characteristics of airborne bacteria should be considered when studying their activities and roles in the atmospheric environment.

Decrease of bioaerosols in westerlies from Chinese coast to the northwestern Pacific: Case data comparisons.

The dissemination of bioaerosols in the westerly wind from the Asian continent to the northwestern Pacific constantly links the land and marine ecosystems. Several observation campaigns targeting bioaerosols were conducted in the coastal city Qingdao of China (QD), at a coast site of Kumamoto in southwestern Japan (KM), and in the northwestern Pacific (NP) between 2014 and 2016. We compared the concentration of bioaerosols in the range of 1.1-7.0 µm obtained in those campaigns to investigate their variation in the westerly wind. The substantial influence of westerlies on bioaerosol concentration was confirmed in the three areas. In the case of non-dust air, the arrival of the continental air led to a 29 % decrease of bioaerosols at KM while a 57 % increase at NP, indicating that the concentration in non-dust air was lower than the local level in the island air while higher than that in the remote marine air. In case of dust occurrence, bioaerosols in the air decreased with the distance from the Asian continent at KM and NP consecutively, and the arrival of the air caused a 2-fold increase at KM and a 1.7-fold increase at NP. The relative concentration increase rate of bioaerosols (IRRC), defined as the ratio of the increment of bioaerosols caused by long-distance transported air to the local level in each area, decreased rapidly after the air left the continent in the dust cases, which is similar to the decrease of the dry deposition flux of dust reported in the literature. This result indicates that the reduction of bioaerosols in the dusty air was likely dominated by the removal of bioaerosols attached to dust particles.

Complete genome sequence of Lactococcus lactis IO-1, a lactic acid bacterium that utilizes xylose and produces high levels of L-lactic acid.

We report the complete genome sequence of Lactococcus lactis IO-1 (= JCM7638). It is a nondairy lactic acid bacterium, produces nisin Z, ferments xylose, and produces predominantly L-lactic acid at high xylose concentrations. From ortholog analysis with other five L. lactis strains, IO-1 was identified as L. lactis subsp. lactis.

Identification of functional clock-controlled elements involved in differential timing of Per1 and Per2 transcription.

It has been proposed that robust rhythmic gene expression requires clock-controlled elements (CCEs). Transcription of Per1 was reported to be regulated by the E-box and D-box in conventional reporter assays. However, such experiments are inconclusive in terms of how the CCEs and their combinations determine the phase of the Per1 gene. Whereas the phase of Per2 oscillation was found to be the most delayed among the three Period genes, the phase-delaying regions of the Per2 promoter remain to be determined. We therefore investigated the regulatory mechanism of circadian Per1 and Per2 transcription using an in vitro rhythm oscillation-monitoring system. We found that the copy number of the E-box might play an important role in determining the phase of Per1 oscillation. Based on real-time bioluminescence assays with various promoter constructs, we provide evidence that the non-canonical E-box is involved in the phase delay of Per2 oscillation. Transfection experiments confirmed that the non-canonical E-box could be activated by CLOCK/BMAL1. We also show that the D-box in the third conserved segment of the Per2 promoter generated high amplitude. Our experiments demonstrate that the copy number and various combinations of functional CCEs ultimately led to different circadian phases and amplitudes.

Regular feeding plays an important role in cholesterol homeostasis through the liver circadian clock.

RATIONALE: Peripheral clock control and the relevance of the circadian rhythm to physiology and disease are major questions in mammalian circadian biology. OBJECTIVE: We examined the physiological functions of the liver clock. METHODS AND RESULTS: We established a suppressed feeding schedule regimen constituting a high-cholesterol diet delivered every 6 hours without changes in energy and cholesterol intake. We found that rats exposed to this regimen developed hypercholesteremia. In the liver, the rhythmicity of expression of several clock genes was disrupted. Furthermore, the nocturnal expression of the CYP7A1 gene, which encodes the rate-limiting enzyme for the conversion of cholesterol to bile acids, was shifted to a diurnal pattern. Indeed, suppression of a regular feeding rhythm increased the secretion rate of very-low-density lipoprotein cholesterol from the liver and decreased the excretion of fecal bile acids. CONCLUSIONS: Our results demonstrated that not only the amount and quality of food but also the timing of meals has crucial health implications.

CERBERUS, a novel U-box protein containing WD-40 repeats, is required for formation of the infection thread and nodule development in the legume-Rhizobium symbiosis.

Endosymbiotic infection of legume plants by Rhizobium bacteria is initiated through infection threads (ITs) which are initiated within and penetrate from root hairs and deliver the endosymbionts into nodule cells. Despite recent progress in understanding the mutual recognition and early symbiotic signaling cascades in host legumes, the molecular mechanisms underlying bacterial infection processes and successive nodule organogenesis are still poorly understood. We isolated a novel symbiotic mutant of Lotus japonicus, cerberus, which shows defects in IT formation and nodule organogenesis. Map-based cloning of the causal gene allowed us to identify the CERBERUS gene, which encodes a novel protein containing a U-box domain and WD-40 repeats. CERBERUS expression was detected in the roots and nodules, and was enhanced after inoculation of Mesorhizobium loti. Strong expression was detected in developing nodule primordia and the infected zone of mature nodules. In cerberus mutants, Rhizobium colonized curled root hair tips, but hardly penetrated into root hair cells. The occasional ITs that were formed inside the root hair cells were mostly arrested within the epidermal cell layer. Nodule organogenesis was aborted prematurely, resulting in the formation of a large number of small bumps which contained no endosymbiotic bacteria. These phenotypic and genetic analyses, together with comparisons with other legume mutants with defects in IT formation, indicate that CERBERUS plays a critical role in the very early steps of IT formation as well as in growth and differentiation of nodules.

[Evaluation of a program to promote network building between disciplinary agencies and informal community organizations: trial in a community comprehensive support center].

OBJECTIVE: To examine the effectiveness of a program promoting network building between disciplinary agencies and informal community organizations (IGOs) comprising community residents, by implemention with staff of a community comprehensive support center (CJCSG). METHODS: The program was implemented for nine staff of a GGSG in Setagaya Ward, Tokyo for a year. For process evaluation, items were assessed concerning the contents of the program such as satisfaction and understandability, participants' goal attainment level at each period of the program, and program satisfaction as a whole. Outcome evaluation included measurement of participants' self-efficacy regarding network building with ICOs before and after the program, using interviews of the members who completed the program. RESULT: Eight out of the nine participants completed the program. All positively evaluated the contents of the program and their own goal attainment at each period of the program. After its completion, they felt highly satisfied. Moreover, there was an improvement in the cognition of the participants, including self-efficacy on network building with IGOs and the atmosphere in the GGSG with regard to network building. CONCLUSION: The efficacy of this program could be confirmed as demonstrated by the staff of the CCSC, although a more detailed assessment of validity and effectiveness will be necessary in the future.

[Program development to promote network building between disciplinary agencies and informal community organizations].

OBJECTIVE: To develop a program that promotes network building between disciplinary agencies and informal community organizations (RIO) comprising community residents. METHODS: Questionnaire and interview surveys were conducted for the staff of a community comprehensive support center in order to assess the needs regarding network building with ICOs. Then, a program was developed based on the social cognitive theory. The aim of the program was to raise awareness about network building and to develop necessary skills. RESULTS: The program induded ten sessions, with themes encompassing "reviewing the past activities", "understanding the significance of network building with IGOs", "integrating the aims and visions of staff", and "learning ways to build networks with IGOs". Objectives of each session were set on the basis of the particular theme covered. Participants performed group work during the sessions. CONCLUSION: Establishment and actual implementation of programs in clinical practice could be expected to strengthen networking between disciplinary agencies and ICOs. It is necessary to implement programs and evaluate their effectiveness in the future.

Different chain length specificity among three polyphosphate quantification methods.

Polyphosphate is ubiquitous among living organisms and has a variety of biochemical functions. Arbuscular mycorrhizal fungi have been known to accumulate polyphosphate as a key compound for their function. However, an enzymatic assay using polyphosphate kinase (PPK) reverse reaction, in which polyphosphate is converted to adenosine triphosphate (ATP) and quantified by luciferase assay, failed to detect accumulation of polyphosphate in some mycorrhizal root. When yeast exopolyphosphatase (PPX) was applied to these samples, a much higher polyphosphate level was detected than when the PPK assay was applied. Detailed analysis of substrate chain length specificity of these methods using polyphosphate chain length standards revealed that the PPX method was the most appropriate to detect short-chain polyphosphate. The average chain length of the shortest polyphosphate fraction that could be quantified with more than 50% efficiency was 3 for the PPX method and 38 for the PPK method. It was also suggested that the ratio of the PPK value to the PPX value may be useful as a simple and relative index to compare polyphosphate chain length distribution in different samples.

Genetics of symbiosis in Lotus japonicus: recombinant inbred lines, comparative genetic maps, and map position of 35 symbiotic loci.

Development of molecular tools for the analysis of the plant genetic contribution to rhizobial and mycorrhizal symbiosis has provided major advances in our understanding of plant-microbe interactions, and several key symbiotic genes have been identified and characterized. In order to increase the efficiency of genetic analysis in the model legume Lotus japonicus, we present here a selection of improved genetic tools. The two genetic linkage maps previously developed from an interspecific cross between L. japonicus Gifu and L. filicaulis, and an intraspecific cross between the two ecotypes L. japonicus Gifu and L. japonicus MG-20, were aligned through a set of anchor markers. Regions of linkage groups, where genetic resolution is obtained preferentially using one or the other parental combination, are highlighted. Additional genetic resolution and stabilized mapping populations were obtained in recombinant inbred lines derived by a single seed descent from the two populations. For faster mapping of new loci, a selection of reliable markers spread over the chromosome arms provides a common framework for more efficient identification of new alleles and new symbiotic loci among uncharacterized mutant lines. Combining resources from the Lotus community, map positions of a large collection of symbiotic loci are provided together with alleles and closely linked molecular markers. Altogether, this establishes a common genetic resource for Lotus spp. A web-based version will enable this resource to be curated and updated regularly.

Notch signaling may be involved in the abnormal differentiation of epidermal keratinocytes in psoriasis.

Localization of each keratin isoform differs among epidermal layers. Proliferating basal cells synthesize keratin 14 (K14) and suprabasal cells express keratin 10 (K10) in normal skin. Notch signaling is essential for keratinocyte differentiation. Notch1 is expressed in all epidermal layers, Notch2 in the basal cell layer and Notch3 in basal cell and spinous cell layers in normal epidermis. It has been poorly elucidated how localization and expression levels of Notch molecules are related to epidermal molecular markers K10 and K14 in psoriatic skin with abnormal differentiation of epidermal tissue. This study aimed to investigate the relationship between abnormal differentiation of epidermal cells in psoriatic skin and expression of Notch molecules. We investigated keratins (K14 and K10) and Notches (1, 2, 3 and 4) using immunohistochemistry in psoriatic skin (n=30) and normal skin (n=10). In normal skin, K14 and K10 were discretely observed in the basal cell layer and suprabasal layer, respectively. In psoriatic skin, K14 was expressed in the pan epidermal layer while it and K10 were co-expressed in some middle suprabasal layer cells. Notch1, 2, 3, and 4 localized in all epidermal layers in normal skin. In psoriatic skin, Notch1, 2, and 4 mainly localized in suprabasilar layers and Notch3 is lacalized in pan epidermal, suprabasilar, and basilar layers. Protein and mRNA of Notch1, 2, and 3 isoforms decreased in psoriatic epidermis compared with normal epidermis. These data suggest that decrements in these Notch molecules might cause aberrant expression of K10 and K14 leading to anomalous differentiation of the epidermis in psoriatic lesions.

Complete genome sequence of Enterococcus mundtii QU 25, an efficient L-(+)-lactic acid-producing bacterium.

Enterococcus mundtii QU 25, a non-dairy bacterial strain of ovine faecal origin, can ferment both cellobiose and xylose to produce l-lactic acid. The use of this strain is highly desirable for economical l-lactate production from renewable biomass substrates. Genome sequence determination is necessary for the genetic improvement of this strain. We report the complete genome sequence of strain QU 25, primarily determined using Pacific Biosciences sequencing technology. The E. mundtii QU 25 genome comprises a 3 022 186-bp single circular chromosome (GC content, 38.6%) and five circular plasmids: pQY182, pQY082, pQY039, pQY024, and pQY003. In all, 2900 protein-coding sequences, 63 tRNA genes, and 6 rRNA operons were predicted in the QU 25 chromosome. Plasmid pQY024 harbours genes for mundticin production. We found that strain QU 25 produces a bacteriocin, suggesting that mundticin-encoded genes on plasmid pQY024 were functional. For lactic acid fermentation, two gene clusters were identified-one involved in the initial metabolism of xylose and uptake of pentose and the second containing genes for the pentose phosphate pathway and uptake of related sugars. This is the first complete genome sequence of an E. mundtii strain. The data provide insights into lactate production in this bacterium and its evolution among enterococci.

Long-term efficacy of psoriasis vulgaris treatments: analysis of treatment with topical corticosteroid and/or vitamin D3 analog, oral cyclosporin, etretinate and phototherapy over a 35-year period, 1975-2010.

Various therapies have been tried for psoriasis. In Japan, biologics began to be used for psoriasis treatment in January 2010. Their clinical efficacy is well known, but biologics cannot be used in all psoriasis patients for reasons such as side-effects and cost. It is necessary to evaluate the effect of long-term psoriasis treatment, but there have been no reports evaluating long-term treatment. Therefore, the outcomes of patients who had been treated at the Tokai University Hospital for more than 5 years, before biological agents were released, were examined. Three categories, classified by initial severity, changes in severity by method of treatment and background characteristics, were investigated. In conclusion, cases of long-term treatment with a combination of topical corticosteroid and topical vitamin D3 analog or oral cyclosporin were found to be effective therapies. Patients with a history of diabetes mellitus or cardiovascular disease of psoriasis were likely to be treatment resistant.