Dataset on motivations and perceptions regarding teaching as a career among teacher education students.

The Factors Influencing Teaching Choice (FIT-Choice) scale was adapted with permission from the original creators [4]. The adapted Japanese version of the FIT-Choice scale was validated using a sample of 202 teacher education students in Japan. This scale measures 12 motivational and six perceptual constructs, as theorised in the FIT-Choice framework, using a 7-point Likert scale. This dataset enhances our understanding of the motivations and perceptions related to pursuing a teaching career among teacher education students in Japan. The entire raw dataset was included in a CSV file. Readers are referred to the original creators' publications [2] according to an agreement restricting the disclosure of all scale items in subsequent publications by the author.

Associations of sensory sensitivity, pain catastrophizing, and alexithymia with dental anxiety.

We aimed to reveal interrelationships between alexithymia, catastrophic thinking, sensory processing patterns, and dental anxiety among 460 participants who were registrants of a Japanese research company. Measures used were the Modified Dental Anxiety Scale, the Adult Sensory Profile, the Pain Catastrophizing Scale, and the 20-item Toronto Alexithymia Scale. The interrelationships among the constructs were analyzed using structural equation modeling, adjusting for age, gender, and negative dental treatment experience. Data from 428 participants were used in the analyses. Sensory sensitivity and pain catastrophizing were independently associated with anticipatory and treatment-related dental anxiety, while difficulty identifying feelings was not. In the mediation model, sensory sensitivity and pain catastrophizing served as full mediators between difficulty identifying feelings and the dimensions of dental anxiety (indirect effects were between 0.13 and 0.15). The strength of the associations was 0.55 from difficulty identifying feelings to both pain catastrophizing and sensory sensitivity, and between 0.24 and 0.26 to anticipatory and treatment-related dental anxiety. The association between trait-like phenomena, such as alexithymia, and dental anxiety may be mediated by neurophysiological and cognitive factors such as sensory sensitivity and pain catastrophizing. These findings could be crucial for new and innovative interventions for managing dental anxiety.

Edible plant oil (EPO)-consumption activity of the isolate Fusarium keratoplasticum EN01 and other relative Fusarium species.

Edible oil is used in humans' daily lives, and the degradation of edible oil is a key process in sewage water treatment and in compost production from food wastes. In this study, a mixed microbial strain EN00, which showed high edible plant oil (EPO)-consumption activity, was obtained from soil via enrichment cultivation. A fungal strain EN01 was isolated from EN00 and relegated to Fusarium keratoplasticum, based on the nucleotide sequences of the TEF1-α gene. Strain EN01 eliminated more than 90% of hydrophobic compounds from the medium containing 1.0% (w/v) EPO within 10 days at 30 °C. The rate of consumption of EPO by EN01 was comparable with that of EN00, suggesting that EN01 was the main microorganism involved in the EPO-consumption ability of EN00. Strain EN01 efficiently utilized EPO as a sole carbon source. The EPO-consumption rate of EN01 was highest among six tested strains of Fusarium solani species complex (FSSC), while two FSSC strains of F. mori and F. cuneirostrum, whose phylogenetic relationships were relatively distant from EN01, had little EPO-eliminating activity. This data implies that the potent EPO-eliminating activity is not general in FSSC strains but is restricted to selected members of this complex. EN01 showed good growth at 25-30 °C, in media with an initial pH of 4-10, and in the presence of 0-3% (w/v) sodium chloride. Although the safety including pathogenicity must be strictly evaluated, some FSSC strains including EN01 have potentials for use in the degradation and elimination of edible oil.

Crystal structure of the GH-46 subclass III chitosanase from Bacillus circulans MH-K1 in complex with chitotetraose.

BACKGROUND: Chitosanases (EC 3.2.1.132) hydrolyze chitosan which is a polymer of glucosamine (GlcN) linked by ß - 1,4 bonds, and show cleavage specificity against partially acetylated chitosan containing N-acetylglucosamine (GlcNAc) residues. Chitosanases' structural underpinnings for cleavage specificity and the conformational switch from open to closed structures are still a mystery. METHODS: The GH-46 subclass III chitosanase from Bacillus circulans MH-K1 (MH-K1 chitosanase), which also catalyzes the hydrolysis of GlcN-GlcNAc bonds in addition to GlcN-GlcN, has had its chitotetraose [(GlcN)4]-complexed crystal structure solved at 1.35 Å resolution. RESULTS: The MH-K1 chitosanase's (GlcN)4-bound structure has numerous structural similarities to other GH-46 chitosanases in terms of substrate binding and catalytic processes. However, subsite -1, which is absolutely specific for GlcN, seems to characterize the structure of a subclass III chitosanase due to its distinctive length and angle of a flexible loop. According to a comparison of the (GlcN)4-bound and apo-form structures, the particular binding of a GlcN residue at subsite -2 through Asp77 causes the backbone helix to kink, which causes the upper- and lower-domains to approach closely when binding a substrate. CONCLUSIONS: Although GH-46 chitosanases vary in the finer details of the subsites defining cleavage specificity, they share similar structural characteristics in substrate-binding, catalytic processes, and potentially in conformational change. GENERAL SIGNIFICANCE: The precise binding of a GlcN residue to the -2 subsite is essential for the conformational shift that occurs in all GH-46 chitosanases, as shown by the crystal structures of the apo- and substrate-bound forms of MH-K1 chitosanase.

Psychometric evaluation of the Japanese version of the fear of pain questionnaire-III and its association with dental anxiety: a cross-sectional study.

BACKGROUND: Fear of pain is a significant concern related to chronic pain and its impact on daily functioning. It is also associated with dental anxiety, highlighting its relevance in dental practice. This study aimed to validate the Japanese version of the Fear of Pain Questionnaire-III (FPQ-III) and explore its relationship with dental anxiety. METHODS: 400 participants completed the Japanese version of the FPQ-III, with 100 participants re-evaluated after one month. Convergent validity was tested against dental anxiety and pain catastrophizing, while discriminant validity was assessed by examining general anxiety and depression correlations. Confirmatory factor analysis was used to examine the factorial validity of the FPQ-III and a shortened version of the FPQ-III (FPQ-9). Item response theory was applied for each subscale to estimate the discriminative power of each item and draw a test information curve. Structural equation modeling (SEM) was used to investigate the relationship between fear of pain and dental anxiety. RESULTS: Data from 400 participants (200 women, 44.9 ± 14.5 years) were analyzed. The FPQ-III showed good internal validity, intra-examiner reliability, discriminant validity, and convergent validity. Confirmatory factor analysis results supported a three-factor structure, and the FPQ-9 showed a good fit. Test information curves demonstrated that the FPQ-9 maintained high accuracy over a similarly wide range as the FPQ-III. SEM revealed that fear of minor pain was associated with dental anxiety via fear of medical pain even in individuals without painful medical or dental experiences (indirect effect 0.48 [95% CI: 0.32-0.81]). Fear of severe pain tended to be higher in individuals with chronic pain compared to those without (latent mean values 0 vs. 0.27, p = 0.002) and was also associated with dental anxiety via fear of medical pain in women (indirect effect 0.15 [95% CI: 0.01-0.34]). CONCLUSION: The Japanese version of the FPQ-9 demonstrated high reliability and validity, making it a valuable tool in dental clinical and research settings. It provides insights into the fear of pain among individuals with chronic pain and dental anxiety, informing potential intervention strategies.

Barley Cultivar Sarab 1 Has a Characteristic Region on the Thylakoid Membrane That Protects Photosystem I under Iron-Deficient Conditions.

The barley cultivar Sarab 1 (SRB1) can continue photosynthesis despite its low Fe acquisition potential via roots and dramatically reduced amounts of photosystem I (PSI) reaction-center proteins under Fe-deficient conditions. We compared the characteristics of photosynthetic electron transfer (ET), thylakoid ultrastructure, and Fe and protein distribution on thylakoid membranes among barley cultivars. The Fe-deficient SRB1 had a large proportion of functional PSI proteins by avoiding P700 over-reduction. An analysis of the thylakoid ultrastructure clarified that SRB1 had a larger proportion of non-appressed thylakoid membranes than those in another Fe-tolerant cultivar, Ehimehadaka-1 (EHM1). Separating thylakoids by differential centrifugation further revealed that the Fe-deficient SRB1 had increased amounts of low/light-density thylakoids with increased Fe and light-harvesting complex II (LHCII) than did EHM1. LHCII with uncommon localization probably prevents excessive ET from PSII leading to elevated NPQ and lower PSI photodamage in SRB1 than in EHM1, as supported by increased Y(NPQ) and Y(ND) in the Fe-deficient SRB1. Unlike this strategy, EHM1 may preferentially supply Fe cofactors to PSI, thereby exploiting more surplus reaction center proteins than SRB1 under Fe-deficient conditions. In summary, SRB1 and EHM1 support PSI through different mechanisms during Fe deficiency, suggesting that barley species have multiple strategies for acclimating photosynthetic apparatus to Fe deficiency.

Physiological and genomic analyses of cobalamin (vitamin B12)-auxotrophy of Lysobacter auxotrophicus sp. nov., a methionine-auxotrophic chitinolytic bacterium isolated from chitin-treated soil.

A novel bacterium, designated 5-21aT, isolated from chitin-treated upland soil, exhibits methionine (Met) auxotrophy and chitinolytic activity. A physiological experiment revealed the cobalamin (synonym, vitamin B12)(Cbl)-auxotrophic property of strain 5-21aT. The newly determined complete genomic sequence indicated that strain 5-21aT possesses only the putative gene for Cbl-dependent Met synthase (MetH) and lacks that for the Cbl-independent one (MetE), which implies the requirement of Cbl for Met-synthesis in strain 5-21aT. The set of genes for the upstream (corrin ring synthesis) pathway of Cbl synthesis is absent in the genome of strain 5-21aT, which explains the Cbl-auxotrophy of 5-21aT. This strain was characterized via a polyphasic approach to determine its taxonomic position. The nucleotide sequences of two copies of the 16S rRNA gene of strain 5-21aT indicated the highest similarities to Lysobacter soli DCY21T(99.8 and 99.9 %) and Lysobacter panacisoli CJ29T(98.7 and 98.8 %, respectively), whose Cbl-auxotrophic properties were revealed in this study. The principal respiratory quinone was Q-8. The predominant cellular fatty acids were iso-C15:0, iso-C16:0 and iso-C17:1 ω9c. The complete genome sequence of strain 5-21aT revealed that the genome size was 4 155 451 bp long and the G+C content was 67.87 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain 5-21aT and its most closely phylogenetic relative L. soli DCY21T were 88.8 and 36.5%, respectively. Based on genomic, chemotaxonomic, phenotypic and phylogenetic data, strain 5-21aT represents a novel species in the genus Lysobacter, for which the name Lyobacter auxotrophicus sp. nov. is proposed. The type strain is 5-21aT (=NBRC 115507T=LMG 32660T).

Effects of Nodulation on Metabolite Concentrations in Xylem Sap and in the Organs of Soybean Plants Supplied with Different N Forms.

The effects of nodulation on N metabolism in soybean plants supplied with various forms of N are not fully understood. Ureides are the principal forms of N transported from nodules, but nitrate and asparagine are the primary N compounds transported from roots supplied with NO3-. In this research, the effects of 1-day treatments of NO3-, NH4+, urea, or NO3- + NH4+ on N metabolite concentrations in xylem sap and each organ were compared between nodulated and non-nodulated soybeans. Capillary electrophoresis and colorimetry were used for the analysis. In the xylem sap of the nodulated plants with an N-free solution, ureides were the major N metabolites, followed by asparagine and glutamine. Ureides concentrations were much lower in the xylem sap of the non-nodulated soybeans. In the NO3- treatment, the concentrations of ureides in the xylem sap of the nodulated plants decreased compared to the control plants. In the NH4+, urea, and NO3- + NH4+ treatments, the concentrations of asparagine and glutamine increased significantly compared with the control and NO3- treatments. Similar changes with the N treatments were observed between the nodulated and non-nodulated soybeans, suggesting that nodulation does not have significant effects on the metabolism of absorbed N in roots.

Evaluation of a Capillary Microsampling Device for Analyzing Plasma Lenvatinib Concentration in Patients With Hepatocellular Carcinoma.

BACKGROUND: The anticancer drug, Lenvima (lenvatinib), has severe side effects. Therapeutic drug monitoring helps ensure its efficacy and safety. Regular and optimally timed blood sampling is tough, especially when lenvatinib is self-medicated. Microsampling using the easy to handle Microsampling Wing (MSW) may help circumvent this problem. However, current lenvatinib detection methods are not sensitive enough to detect its concentrations in microsamples (<50-250 µL). Thus, the aim of this study was 2-fold (1) develop an analytic method to estimate plasma lenvatinib concentrations in microsamples and (2) verify whether this method works on micro (5.6 µL) blood plasma samples obtained clinically through MSW from patients with unresectable hepatocellular carcinoma (HCC). METHODS: A simple, highly sensitive, and specific liquid chromatography-electrospray ionization tandem mass spectrometry method was developed. Using this novel protocol, the trough blood plasma concentration of lenvatinib was measured for both blood sampled conventionally and that using MSW. Thirty-five venous whole blood samples were obtained from 11 patients with HCC. Furthermore, the stability of lenvatinib in MSW samples during storage was evaluated. RESULTS: The mean plasma lenvatinib concentration estimates were not significantly different between the MSW and conventional venous blood samples. CV for interday and intraday assays was low. Up to day 5, the lenvatinib concentration in the MSW samples was 85%-115% of the initial day concentration (when stored at 25°C or 4°C). The interference of endogenous matrix components in the human plasma was low. CONCLUSIONS: These results indicate that the novel mass spectrometry protocol accurately measures lenvatinib in human plasma and is reproducible. Thus, MSW could be a useful microsampling device for lenvatinib therapeutic drug monitoring in patients with HCC when used in combination with this novel liquid chromatography-electrospray ionization tandem mass spectrometry detection method.

Shotgun proteomic investigation of methyltransferase and methylation profiles in lipopolysaccharide stimulated RAW264.7 murine macrophages.

Arginine methylation is a common post-translational modification which functions as an epigenetic regulator of transcription and plays a key role in various cell signaling pathways. The methylation of arginine residues is catalyzed by protein arginine methyltransferase (PRMT). However, the expression pattern and underlying mechanism of PRMTs and protein methylation profile in lipopolysaccharide (LPS)-induced innate immune responses are poorly understood. Using a shotgun proteomic approach, we found that LPS stimulation increased arginine and proline metabolism and responses to inflammation and bacterial infections. In comparison, cysteine and methionine metabolism, the pentose phosphate pathway, purine metabolism, and protein methylation factors were also decreased in LPS stimulated murine macrophage cell lines. We revealed that LPS stimulation downregulated PRMT1, PRMT5, and protein arginine methylation profiles in RAW264.7 cells using western blot analysis. Additionally, this phenomenon occurred in parallel with nitric oxide accumulation in LPS-induced macrophages. Using inflammation models, we demonstrate for the first time that LPS stimulation decreases PRMTs, leading to the decreasing of arginine methylation in macrophages.

Prevalence of saliva immunoglobulin A antibodies reactive with severe acute respiratory syndrome coronavirus 2 among Japanese people unexposed to the virus.

While the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a threat to public health as the number of cases and COVID-19-related deaths are increasing worldwide, the incidence of the virus infection is extremely low in Japan compared with many other countries. To explain this uncommon phenomenon, we investigated the prevalence of naturally occurring ("natural") antibodies, focusing on those of the secretory immunoglobulin A (sIgA) form, reactive with SARS-CoV-2 among Japanese people. One hundred and eighty healthy Japanese volunteers of a wide range of age who had been considered to be unexposed to SARS-CoV-2 participated in this study. Saliva samples and blood samples were collected from all of the 180 participants and 139 adults (aged ≥ 20 years) included therein, respectively. The determination of saliva IgA antibodies, mostly comprising sIgA antibodies, as well as serum IgA and immunoglobulin G antibodies, reactive with the receptor binding domain of the SARS-CoV-2 spike-1 subunit proteins was conducted using an enzyme-linked immunosorbent assay. The major findings were that 52.78% (95% confidence interval, 45.21%-60.25%) of the individuals who had not been exposed to SARS-CoV-2 were positive for saliva IgA antibodies with a wide range of levels between 0.002 and 3.272 ng/mL, and that there may be a negative trend in positivity for the antibodies according to age. As we had expected, a frequent occurrence of assumable "natural" sIgA antibodies reactive with SARS-CoV-2 among the studied Japanese participant population was observed.

Cicada-Wing-Inspired Nanopillar Hydrogels Consisting of Poly(vinyl alcohol) and Poly(methacrylic acid) for Capturing Bacteria through Their Flexibility and Wide Range of Motion.

The human race has learned about countering bacterial infection from other living organisms. In this study, cicada-wing-inspired hydrophilic nanopillars were prepared using commercially available porous alumina templates without the use of specialized equipment and additional materials. Hydrophilic polymers of poly(vinyl alcohol) (PVA) and poly(methacrylic acid) (poly(MAAc)) were used to construct these nanopillars. The water-insoluble nanopillars formed by thermal cross-linking between the hydroxy and carboxy groups were highly flexible, with no significant structural disruption being observed on tilting them. The nanopillars captured Gram-negative and Gram-positive bacteria, despite their negatively charged surface leading an electrostatic repulsion with bacteria. The wing of the cicada has evolved to remain dry and maintain its lightweight. The water-containing cicada-wing structure may be an unknown encounter for bacteria and, thus, may become a countermeasure against the ever-changing bacteria.

"Live-Autoradiography" Technique Reveals Genetic Variation in the Rate of Fe Uptake by Barley Cultivars.

Iron (Fe) is an essential trace element in plants; however, the available Fe in soil solution does not always satisfy the demand of plants. Genetic diversity in the rate of Fe uptake by plants has not been broadly surveyed among plant species or genotypes, although plants have developed various Fe acquisition mechanisms. The "live-autoradiography" technique with radioactive 59Fe was adopted to directly evaluate the uptake rate of Fe by barley cultivars from a nutrient solution containing a very low concentration of Fe. The uptake rate of Fe measured by live autoradiography was consistent with the accumulation of Fe-containing proteins on the thylakoid membrane. The results revealed that the ability to acquire Fe from the low-Fe solution was not always the sole determinant of tolerance to Fe deficiency among barley genotypes. The live-autoradiography system visualizes the distribution of ß-ray-emitting nuclides and has flexibility in the shape of the field of view. This technique will strongly support phenotyping with regard to the long-distance transport of nutrient elements in the plant body.

Application of Nitrate, Ammonium, or Urea Changes the Concentrations of Ureides, Urea, Amino Acids and Other Metabolites in Xylem Sap and in the Organs of Soybean Plants (Glycine max (L.) Merr.).

Soybean (Glycine max (L.) Merr.) plants form root nodules and fix atmospheric dinitrogen, while also utilizing the combined nitrogen absorbed from roots. In this study, nodulated soybean plants were supplied with 5 mM N nitrate, ammonium, or urea for 3 days, and the changes in metabolite concentrations in the xylem sap and each organ were analyzed. The ureide concentration in the xylem sap was the highest in the control plants that were supplied with an N-free nutrient solution, but nitrate and asparagine were the principal compounds in the xylem sap with nitrate treatment. The metabolite concentrations in both the xylem sap and each organ were similar between the ammonium and urea treatments. Considerable amounts of urea were present in the xylem sap and all the organs among all the treatments. Positive correlations were observed between the ureides and urea concentrations in the xylem sap as well as in the roots and leaves, although no correlations were observed between the urea and arginine concentrations, suggesting that urea may have originated from ureide degradation in soybean plants, possibly in the roots. This is the first finding of the possibility of ureide degradation to urea in the underground organs of soybean plants.

Enhancement of Photosynthetic Iron-Use Efficiency Is an Important Trait of Hordeum vulgare for Adaptation of Photosystems to Iron Deficiency.

Leaf iron (Fe) contents in Fe-deficiency-tolerant plants are not necessarily higher than that in Fe-deficiency-susceptible ones, suggesting an unknown mechanism involved in saving and allowing the efficient use of minimal Fe. To quantitatively evaluate the difference in Fe economy for photosynthesis, we compared the ratio of CO2 assimilation rate to Fe content in newly developed leaves as a novel index of photosynthetic iron-use efficiency (PIUE) among 23 different barley (Hordeum vulgare L.) varieties. Notably, varieties originating from areas with alkaline soil increased PIUE in response to Fe-deficiency, suggesting that PIUE enhancement is a crucial and genetically inherent trait for acclimation to Fe-deficient environments. Multivariate analyses revealed that the ability to increase PIUE was correlated with photochemical quenching (qP), which is a coefficient of light energy used in photosynthesis. Nevertheless, the maximal quantum yield of photosystem II (PSII) photochemistry, non-photochemical quenching, and quantum yield of carbon assimilation showed a relatively low correlation with PIUE. This result suggests that the ability of Fe-deficiency-tolerant varieties of barley to increase PIUE is related to optimizing the electron flow downstream of PSII, including cytochrome b6f and photosystem I.

Transition of the Bacterial Community and Culturable Chitinolytic Bacteria in Chitin-treated Upland Soil: From Streptomyces to Methionine-auxotrophic Lysobacter and Other Genera.

Chitin amendment is an agricultural management strategy for controlling soil-borne plant disease. We previously reported an exponential decrease in chitin added to incubated upland soil. We herein investigated the transition of the bacterial community structure in chitin-degrading soil samples over time and the characteristics of chitinolytic bacteria in order to elucidate changes in the chitinolytic bacterial community structure during chitin degradation. The addition of chitin to soil immediately increased the population of bacteria in the genus Streptomyces, which is the main decomposer of chitin in soil environments. Lysobacter, Pseudoxanthomonas, Cellulosimicrobium, Streptosporangium, and Nonomuraea populations increased over time with decreases in that of Streptomyces. We isolated 104 strains of chitinolytic bacteria, among which six strains were classified as Lysobacter, from chitin-treated soils. These results suggested the involvement of Lysobacter as well as Streptomyces as chitin decomposers in the degradation of chitin added to soil. Lysobacter isolates required yeast extract or casamino acid for significant growth on minimal agar medium supplemented with glucose. Further nutritional analyses demonstrated that the six chitinolytic Lysobacter isolates required methionine (Met) to grow, but not cysteine or homocysteine, indicating Met auxotrophy. Met auxotrophy was also observed in two of the five type strains of Lysobacter spp. tested, and these Met auxotrophs used d-Met as well as l-Met. The addition of Met to incubated upland soil increased the population of Lysobacter. Met may be a factor increasing the population of Lysobacter in chitin-treated upland soil.

Effects of Different Chemical Forms of Nitrogen on the Quick and Reversible Inhibition of Soybean Nodule Growth and Nitrogen Fixation Activity.

It has been reported that supply of nitrate to culture solution rapidly and reversibly inhibits nodule growth and nitrogen fixation activity of soybean. In this study, the effects of ammonium, urea, or glutamine on nodule growth and nitrogen fixation activity are compared with that for nitrate. Soybean plants were cultivated with a nitrogen-free nutrient solution, then 1 mM-N of nitrate, ammonium, glutamine, or urea were supplied from 12 DAP until 17 DAP. Repression of nodule growth and nitrogen fixation activity at 17 DAP were observed by ammonium, urea, and glutamine like nitrate, although the inhibitory effects were milder than nitrate. The removal of nitrogen from the culture solutions after nitrogen treatments resulted in a recovery of the nodule growth. It was found that the glutamine treatment followed by N-free cultivation gave highest nitrogen fixation activity about two times of the control. Tracer experiments with 15N and 13C were performed to evaluate the translocation of N and C to the different tissues. Culture solutions containing a 15N-labeled nitrogen source were supplied from 21 DAP, and the whole shoots were exposed to 13CO2 for 60 min on 23 DAP, and plants were harvested on 24 DAP. The percentage distribution of 15N in nodules was highest for ammonium (1.4%) followed by glutamine (0.78%), urea (0.32%) and nitrate (0.25%). The percentage distribution of 13C in the nodules was highest for the control (11.5%) followed by urea (5.8%), glutamine (2.6%), ammonium (2.3%), and nitrate (2.3%). The inhibitory effects of nitrogen compounds appeared to be related to a decrease in photoassimilate partitioning in the nodules, rather than 15N transport into the nodules. The free amino acid concentrations after nitrogen treatments were increased in the nodules and leaves by nitrate, in the roots by ammonium, in the stems by urea, and the roots, stems, and leaves by glutamine treatment. The concentrations of asparagine, aspartate, and glutamine were increased after nitrogen treatments. By the long-term supply of nitrogen for 2-weeks, nitrate significantly increased the lateral roots and leaf growth. The long-term supply of urea and glutamine also promoted the lateral roots and leaf growth, but ammonium suppressed them.

A rice PHD-finger protein OsTITANIA, is a growth regulator that functions through elevating expression of transporter genes for multiple metals.

Essential metal absorption for plant growth is mediated predominantly by metal-specific transporters, with expression that responds to the environmental or cellular conditions of specific metals. Differing from metal-specific regulation, we describe a constitutively expressed transcription factor that regulates the transport of several metals in rice. We characterized the rice mutant LOW CADMIUM 5 (LC5), which exhibited reduced growth and accumulation of essential metals (e.g., copper [Cu], zinc [Zn] and manganese [Mn]) in shoots. LC5 was dwarf and developed less tillers than the wild type, but the structure of vasculature was apparently normal. Molecular genetic analysis revealed that the causal gene of LC5 is an ortholog of the transcriptional regulator Arabidopsis thaliana TITANIA (TTA), known as a transcriptional regulator. Expression analyses demonstrated that the OsTTA gene encodes a nucleus-localized protein containing a plant homeodomain-finger (PHD-finger) domain and is expressed ubiquitously in rice plants. RNA sequencing and quantitative PCR analyses revealed that the mRNA accumulation of transporter genes for essential metals, including iron (Fe), Zn, or Mn, were substantially lower in LC5 roots than in the wild type. Unlike known transcription factors of metal transport regulation, OsTTA transcript accumulation was not affected by metal availability. In addition, the growth defect of LC5 was partially rescued by Fe, Zn, or Mn supplementation, respectively. Taken together, OsTTA is a constitutively expressed regulator of multiple metal transporter genes responsible for essential metals delivery to shoots for their normal growth.

NgcESco Acts as a Lower-Affinity Binding Protein of an ABC Transporter for the Uptake of N,N'-Diacetylchitobiose in Streptomyces coelicolor A3(2).

In the model species Streptomyces coelicolor A3(2), the uptake of chitin-degradation byproducts, mainly N,N'- diacetylchitobiose ([GlcNAc]2) and N-acetylglucosamine (GlcNAc), is performed by the ATP-binding cassette (ABC) transporter DasABC-MsiK and the sugar-phosphotransferase system (PTS), respectively. Studies on the S. coelicolor chromosome have suggested the occurrence of additional uptake systems of GlcNAc-related compounds, including the SCO6005-7 cluster, which is orthologous to the ABC transporter NgcEFG of S. olivaceoviridis. However, despite conserved synteny between the clusters in S. coelicolor and S. olivaceoviridis, homology between them is low, with only 35% of residues being identical between NgcE proteins, suggesting different binding specificities. Isothermal titration calorimetry experiments revealed that recombinant NgcESco interacts with GlcNAc and (GlcNAc)2, with Kd values (1.15 and 1.53 µM, respectively) that were higher than those of NgcE of S. olivaceoviridis (8.3 and 29 nM, respectively). The disruption of ngcESco delayed (GlcNAc)2 consumption, but did not affect GlcNAc consumption ability. The ngcESco-dasA double mutation severely decreased the ability to consume (GlcNAc)2 and abolished the induction of chitinase production in the presence of (GlcNAc)2, but did not affect the GlcNAc consumption rate. The results of these biochemical and reverse genetic analyses indicate that NgcESco acts as a (GlcNAc)2- binding protein of the ABC transporter NgcEFGSco-MsiK. Transcriptional and biochemical analyses of gene regulation demonstrated that the ngcESco gene was slightly induced by GlcNAc, (GlcNAc)2, and chitin, but repressed by DasR. Therefore, a model was proposed for the induction of the chitinolytic system and import of (GlcNAc)2, in which (GlcNAc)2 generated from chitin by chitinase produced leakily, is mainly transported via NgcEFG-MsiK and induces the expression of chitinase genes and dasABCD.

Simultaneous analysis of multiple urinary biomarkers for the evaluation of oxidative stress by automated online in-tube solid-phase microextraction coupled with negative/positive ion-switching mode liquid chromatography-tandem mass spectrometry.

This study described an automated online method for the simultaneous determination of 8-isoprostane, 8-hydroxy-2'-deoxyguanosine, and 3-nitro-l-tyrosine in human urine. The method involves in-tube solid-phase microextraction using a Carboxen 1006 PLOT capillary column as an extraction device, followed by liquid chromatography with tandem mass spectrometry using a CX column and detection in the negative/positive switching ion-mode by multiple reaction monitoring. Using their stable isotope-labeled internal standards, each of these oxidative stress biomarkers showed good linearity from 0.02 to 2.0 ng/mL. Their detection limits (S/N = 3) were 3.4-21.5 pg/mL, and their intra- and inter-day precisions (relative standard deviations) were >3.9 and 6.5% (n = 5), respectively. This method was applied successfully to the analysis of urine samples, without any other pretreatment and interference peaks.