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RabGTPase is a member of the Ras superfamily of small GTPases, which share a GTP-binding pocket containing highly conserved motifs that promote GTP hydrolysis. In Arabidopsis, the RabA group, which corresponds to the Rab11 group in animals, functions in the recycling of endosomes that control docking and fusion during vesicle transport. However, their molecular mechanisms remain unknown. In this study, we determined the crystal structures of the GDP-bound inactive form and both GppNHp- and GTP-bound active forms of RabA1a, at resolutions of 2.8, 2.6, and 2.6 Å, respectively. A bound sulfate ion in the active site of the GDP-bound structure stabilized Switch II by bridging the interaction between a magnesium ion and Arg74. Comparisons of the two states of RabA1a with Rab11 proteins revealed clear differences in the Switch I and II loops. These results suggested that conformational change of the Switch regions of RabA1a, derived by GTP or GDP binding, could maintain subcellular membrane traffic through the specific interaction of effector molecules.

Isoflavonoids are a class of biologically active natural products that accumulate in soybean ( Glycine max L.) seeds during development, play vital roles in plant defense, and act as phytoestrogens with important human health benefits. Plant cell suspension cultures represent an excellent source of biologically important secondary metabolites. We found that methyl jasmonate (MJ) treatment increased isoflavone production in soybean suspension cell cultures. To investigate the underlying mechanism, we examined the expression of structural genes ( CHS6, CHS7, CHI1, IFS1, IFS2, IFMaT, and HID) in the isoflavonoid biosynthesis pathways in soybean suspension cells under various abiotic stress conditions. MJ treatment had the most significant effect on gene expression and increased the production of three glycosidic isoflavones (daidzin, malonyldaidzin, and malonylgenistin), with the maximum total isoflavone production (∼10-fold increase) obtained on day 9 after MJ application. MJ treatment significantly increased total phenolic contents and upregulated isoflavonoid biosynthesis genes, shedding light on the underlying mechanism.

TAGLN is an actin-binding protein family that comprises three isoforms with theorized roles in smooth muscle differentiation, tumour development, lymphocyte activation, and brain chemistry. However, their fundamental characteristics in regulation of the actin-based cytoskeleton are not fully understood. Here we show that TAGLN2 (including TAGLN1 and TAGLN3) extensively nucleates G-actin polymerization under low-salt conditions, where polymerization would be completely suppressed. The calponin homology domain and actin-binding loop are essential to mechanically connect two adjacent G-actins, thereby mediating multimeric interactions. However, TAGLN2 blocked the Arp2/3 complex binding to actin filaments under physiological salt conditions, thereby inhibiting branched actin nucleation. In HeLa and T cells, TAGLN2 enhanced filopodium-like membrane protrusion. Collectively, the dual functional nature of TAGLN2—G-actin polymerization and Arp2/3 complex inhibition—may account for the mechanisms of filopodia development at the edge of Arp2/3-rich lamellipodia in various cell types.

Sophorae Radix (Sophora flavescens Aiton) has long been used in traditional medicine in East Asia due to the various biological activities of its secondary metabolites. Endogenous contents of phenolic compounds (phenolic acid, flavonol, and isoflavone) and the main bioactive compounds of Sophorae Radix were analyzed based on the qualitative HPLC analysis and evaluated in different organs and at different developmental stages. In total, 11 compounds were detected, and the composition of the roots and aerial parts (leaves, stems, and flowers) was significantly different. trans-Cinnamic acid and p-coumaric acid were observed only in the aerial parts. Large amounts of rutin and maackiain were detected in the roots. Four phenolic acid compounds (benzoic acid, caffeic acid, ferulic acid, and chlorogenic acid) and four flavonol compounds (kaempferol, catechin hydrate, epicatechin, and rutin) were higher in aerial parts than in roots. To identify putative genes involved in phenolic compounds biosynthesis, a total of 41 transcripts were investigated. Expression patterns of these selected genes, as well as the multiple isoforms for the genes, varied by organ and developmental stage, implying that they are involved in the biosynthesis of various phenolic compounds both spatially and temporally.

Introduction: Endovascular recanalization (EVT) for acute ischemic stroke patients with sizable penumbral tissues seems plausible even beyond 6 hours from their last seen normal (LSN), provided that there remain salvageable tissues. Methods: A total of 111 acute ischemic stroke patients were included in the analysis, who had ICA or MCA occlusion cases with baseline NIHSS score ≥6-point and arrived 6 - 12 hours after LSN. Baseline demographics, stroke information and functional outcomes were prospectively collected during routine clinical practice. Attending physicians made treatment decisions for EVT by current guidelines and institutional protocols. MR image parameters, including volume of diffusion-restrcted lesions (ADC value ≤450) and mapping of hypoperfused area (T max delay ≥6 seconds), were quantified using an automated commercial software with minimal manual operations. Binary logistic regression analysis models taking mRS score 0 - 1 at 3 months after stroke as a dependent variable were constructed. Results: Of the 111 ICA or MCA occlusion cases who arrived 6 - 12 hours after LSN, 58% had mismatch ratio of ≥1.8 at baseline and 42% had favorable imaging profiles by DEFUSE-2 study. EVT was done in 54% of cases. Three-month mRS score 0 - 1 was found in 19% and mortality in 14%. EVT was associated with significantly increased odds of having mRS score 0 - 1 at 3 months after stroke (adjusted OR, 6.15 [95% CI, 1.12 - 44.10]) after controlling for covariates. Good response to EVT was dependent upon the baseline infarct volume and time delay (Figure) Sensitivity analyses for propensity weighting, location of occlusion, preceding intravenous tPA, size of penumbral tissues showed consistent associations favoring for EVT. Conclusions: EVT is feasible and efficacious for anterior circulation occlusion with substantial neurological deficits even 6 - 12 hours after LSN. More research is warranted to target the best candidates of EVT using image parameters.

Background: Clinical trials have shown that benefits of endovascular recanalization (EVT) for acute ischemic stroke patients with sizable penumbral tissues seems plausible even beyond 6 h after their last seen normal (LSN). Persistency of ischemic penumbra remains unclear in delayed periods. Methods: From a prospective stroke registry database, we identified 111 acute ischemic stroke patients who had internal carotid artery or middle cerebral artery occlusion with baseline National Institutes of Health Stroke Scale scores ≥6 points and arrived 6–12 h after LSN. Baseline information and functional outcomes were prospectively collected as a clinical registry. Attending physicians made treatment decisions for EVT based on the current guidelines and institutional protocols. MR image parameters, including the volume of diffusion-restricted lesions and mapping of the ­hypoperfused area, were quantified using automated commercial software. Binary logistic regression analysis models, with modified Rankin Scale (mRS) scores of 0–1 at 3 months after stroke included as a dependent variable, were constructed. Results: Between 6 and 12 h after onset, 58% had a mismatch ratio of ≥1.8 at baseline and 42% had favorable imaging profiles as determined by DEFUSE 2 study. After 9 h, there was a mismatch ratio of ≥1.8 in 47 and 38% favorable profiles. EVT was performed in 54% of cases. A 3-month mRS score of 0–1 was found in 19% (25% in EVT and 12% in medical treatment groups) of cases. EVT was associated with an increased OR of having a mRS score of 0–1 at 3 months after stroke (adjusted OR 7.59 [95% CI 1.28–61.60]). Conclusions: Penumbral tissues were persistent in a substantial proportion of anterior circulation occlusion cases 6–12 h after LSN. EVT at 6–12 h in a predominantly Asian cohort resulted in better outcomes.

Figure S2. Schematic illustration of the strategy used to clone BoMYBL2–1 from various B. oleracea species. A: Genomic region around the BoMYBL2–1 based on information obtained from two different databases (upper: http://brassicadb.org/brad ; Liu et al. 2014; lower: http://plants.ensembl.org/Brassica_oleracea/Info/Index ; Parkin et al. 2014). B: Illustration showing the positions of the fragments, amplified using different combinations of primer sets, used to assemble the entire promoter and coding sequence of BoMYBL2–1. The yellow block represents a 159 bp repeat sequence. All primer sets are listed in Table 2. BoLPR2 is B. oleracea multicopper oxidase LPR2; BoPUB10 is B. oleracea U-box domain-containing protein 10. (DOCX 72 kb)

Figure S3. Results of genomic DNA-PCR preformed to detect the presence or absence of genes regulating anthocyanin biosynthesis other than BoMYBL2–1. A: B. oleracea var. capitata F. alba or rubra varieties with contrasting characteristics in anthocyanin accumulation were selected for analysis; varieties with green and purple colors are indicated in green or purple. B: PCR analysis of other varieties of B. oleracea. The color of each variety is indicated above each lane. (DOCX 293 kb)

Figure S5. Comparison of different BoMYBL2–1 nucleotide sequences obtained from cabbages. Shaded regions indicate exon sequences. Sequences corresponding to Bol016162 from B. oleracea var. capitata were omitted. (DOCX 31 kb)

Table S2. List of primer sequences used in RT-PCR and genomic PCR analyses of BoMYBL2. (DOCX 32Â kb)

Figure S1. Total anthocyanin content (A) of samples of the cabbages (B) shown in Fig. 1. (DOCX 109 kb)

Table S1. Primers used to clone BoMYBL2. (DOCX 19Â kb)

Figure S4. Expression of genes associated with anthocyanin biosynthesis in various purple cabbages. Daebakna is a green cabbage used as a reference. (DOCX 387Â kb)

Macrophages are one of the major cell types that produce IL-1β. IL-1β maturation occurs via inflammasome activation, and mature IL-1β is then released from the cell. Secreted IL-1β mediates inflammatory reactions in various pathological environments, such as those in infectious, autoimmune, and cancerous diseases. Although the mechanism of IL-1β production has been discovered in infectious and autoimmune diseases, its production mechanism in the tumor microenvironment is unclear. Therefore, the mechanism of IL-1β production in macrophages in the tumor microenvironment was investigated in this study. First, bone marrow-derived macrophages obtained from C57BL/6 mice were treated with B16F10 tumor-conditioned media (TCM) in vitro. TCM increased the levels of IL-1β via glucose-mediated activation of the inflammasome. Moreover, TCM enhanced the activation of both NF-κB and mTOR pathways in a glucose-dependent manner. In particular, the expression levels of mTORC1 component proteins were dependent on the TCM-induced activation of NF-κB signaling. In addition, TCM affected ASC-ASC interactions through increasing intracellular reactive oxygen species levels. Finally, glucose inhibition by inoculation with 2-deoxy-D-glucose in vivo decreased the IL-1β levels in both the blood and tumor region of B16F10-bearing C57BL/6 mice relative to those in PBS-injected tumor-bearing mice. These results suggest that glucose supplied from blood vessels might be important for IL-1β production in tumor-associated macrophages via the integrated signals of the NF-κB and mTOR pathways in the tumor microenvironment.

In this study, hierarchically three-dimensional (3D) nanotubular sea urchin-shaped iron oxide nanostructures (3D-Fe2O3) were synthesized by a facile and rapid ultrasound irradiation method. Additives, templates, inert gas atmosphere, pH regulation, and other complicated procedures were not required. Dense 3D-Fe2O3 with a relatively large Brunauer–Emmett–Teller (BET) surface area of 129.4 m2/g was synthesized within 23 min, and the BET surface area was further improved to 282.7 m2/g by a post heat-treatment process. In addition, this post processing led to phase changes from maghemite (γ phase) to hematite (α phase) Fe2O3. Subsequent characterization suggested that the growth mechanism of the 3D-Fe2O3 follows self-assembly and oriented attachment. The prepared 3D-Fe2O3 was applied to wastewater purification. Ultrasound-irradiated 3D-Fe2O3 can eliminate a As(V) and Cr(VI) from water with 25 times faster removal rate by using a one third smaller amount than commercial α-Fe2O3. This was attributed to the inter-particle pores and relatively positively charged surface of the nanostructure. In addition, post heat treatment on ultrasound-irradiated 3D-Fe2O3 significantly influenced the photocatalytic degradation of methylene blue and phenol, with a 25 times higher removal efficiency than that of commercial α-Fe2O3, because of both high BET surface area and good crystallization of the prepared samples.

In this paper, it is first reported that gray hydrogenated TiO

Background Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is known to play a role in the pathogenesis of various inflammatory diseases. However, no study has been performed on childhood asthma. Methods Ninety-five children with asthma and 78 controls aged 5–18 years were included. Sputum induction, pulmonary function test (PFT), and methacholine challenge test were performed. The subjects were divided into the eosinophilic airway (EA) and non-EA (NEA) groups based on sputum analysis and into the high and low TWEAK groups according to the TWEAK cutoff level (263.0 pg/mL). TWEAK in induced sputum supernatant was measured through enzyme-linked immunosorbent assay. Results Children with asthma had higher TWEAK levels than healthy controls [493.0 (157.1–904.3) vs. 118.2 (67.5–345.5) pg/mL, P < 0.001]. Sputum TWEAK levels were significantly correlated with PFT parameters reflecting airway obstruction. This association was particularly prominent in subjects with NEA inflammation. Significant differences in FEF25–75 (maximum midexpiratory flow, % predicted; P = 0.017), AX (reactance area; P < 0.001), R5-R20 (difference between resistance at 5 and 20 Hz; P = 0.025), and X5 (reactance at 5 Hz, % predicted; P < 0.001) were noted between the high and low TWEAK groups within the NEA group. Sputum TWEAK level also showed significant positive correlations with asthma severity (r = 0.358, P = 0.001) and control status (r = 0.470, P < 0.001), distinctively in subjects with NEA inflammation. Conclusions Airway TWEAK may play a role in small airway inflammation especially in children with noneosinophilic asthma. This article is protected by copyright. All rights reserved.

This article attempts to develop a practical theology of resistance for religious education. It is inspired by the struggle of indigenous people in Guatemala in their memorialization of the Rio Negro Massacre in Guatemala, celebrations of International Women's Day, and the creation of a school for survivors of the violence surrounding Rio Negro. Remembrance, relationship-building, and reclaiming space will be suggested as practices to be employed by religious education. This article proposes that these three interrelated elements are integral to religious education. It concludes that a theology of resistance equipped with these three aspects promotes and deepens a practice of faith, emunah, to nourish.

The resistance to Diamondback moth insect in cabbage is governed by many minor loci in quantitative nature, and at least four genetic loci should be incorporated in marker-assisted breeding program for developing partially resistant DBM cabbage cultivars. The Diamondback moth (DBM), Plutella xylostella (L.), is the most destructive insect infesting cruciferous plants worldwide. Earlier studies have reported that the glossy leaves of cabbage are associated with resistance to this insect. However, until now, genetics of DBM resistance has not been studied in detail, and no QTL/gene mapping for this trait has been reported. In this paper, we report quantitative trait loci (QTL) mapping of DBM-resistant trait using 188 randomly selected segregating F 3 population derived from crossing a partially DBM-resistant glossy leaf cabbage (748) with a susceptible smooth cabbage line (747). Quantitative trait loci mapping using phenotypic data of four consecutive years (2008, 2009, 2010, and 2011) on DBM insect infestation detected a total of eight QTL on five linkage groups suggesting that DBM resistance is a quantitative in nature. Of these QTL, four QTL, i.e., qDbm 1 on LG1, qDbm5 and qDbm6 on LG7, and qDbm8 on LG9, were detected in different tests and years. The QTL, qDbm6 on LG7, was consecutively detected over 3 years. Tightly linked molecular markers have been developed for qDbm8 QTL on LG9 which could be used in marker-assisted breeding program. Our research demonstrated that for desired DBM resistance cultivar breeding, those four genetic loci have to be taken into consideration. Furthermore, the comparative study revealed that DBM resistance QTL is conserved between close relative model plant Arabidopsis thaliana and Brassica oleracea genome.

The aerial surfaces of plants are covered with a wax layer that serves the essential functions of limiting non-stomatal water loss and acting as protective barrier against environmental stresses. We selected two broccoli lines, bloomed (MC91) and bloomless (MC117), and analyzed their phenotypes related to cuticular wax accumulation. The total wax amount was 1.93-fold higher in MC91 leaves compared to MC117 leaves. All of the studied cuticular wax compounds were 1.07–3.79-fold higher in MC91 plants compared to MC117 plants except for the C 31 alkane. The wax compositions did not essentially different between the two broccoli lines, but some compounds were found at significantly higher levels in MC91 plants compared to MC117 plants, mainly reflecting differences in C 29 alkanes, C 29 secondary alcohols and C 29 ketones. To investigate gene regulation by bloom phenotype, we analyzed the mRNA expression patterns of various cuticular wax biosynthetic genes. Our results revealed that LACS1 , KCS1 , KCR1 , ECR , CER3 and MAH1 were expressed more in MC91 plants compared to MC117 plants at both 3 and 10 weeks. The expression levels of the studied cuticular wax biosynthetic genes were significantly induced by drought stress, which is known to induce cuticular wax deposition. Together, these results show that the cuticular wax accumulation of broccoli is regulated by cuticular wax biosynthetic gene expression and can be affected by environmental signals.

Solution phase post-modification of a trimesic acid (TMA) network on Au(111) with Zn(2+) ions was found to induce transformation of a bar-featured scanning tunneling microscopy image of (5√3 × 5√3) to a chevron-pair image of (10√3 × 10√3). Voltammetric determination of Zn coverage in the modified TMA network supports the fact that the chevron feature consists of three TMA molecules combined via two coordination bonds between Zn(2+) ions and carboxylates of TMA.

To characterize of the pathogenicity gene from the soft rot pathogen Pantoea sp. PPE7 in Pleurotus eryngii , we constructed over 10,000 kanamycin-resistant transposon mutants of Pantoea sp. strain PPE7 by transposon mutagenesis. One mutant, Pantoea sp. NPPE9535, did not cause a soft rot disease on Pleurotus eryngii was confirmed by the pathogenicity test. The transposon was inserted into the wzc gene and the disruption of the wzc gene resulted in the reduction of polysaccharide production and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii . Analysis of the hydropathic profile of this protein indicated that it is composed of two main domains: an N-terminal domain including two transmembrane α-helices and a C-terminal cytoplasmic domain consisting of a tyrosine-rich region. Comparative analysis indicated that the amino acid sequence of Wzc is similar to that of a number of proteins involved in the synthesis or export of polysaccharides in other bacterial species. Purified GST-Wzc was found to affect the phosphorylation of tyrosine residue in vivo . These results showed that the wzc gene might play an important role in the virulence of Pantoea sp. strain PPE7 in P. eryngii .

The present study investigated variations in soil microbial communities and the chemical properties of forest soils by differing amounts of penetrating sunlight. The soil temperature was significantly higher in higher light-penetrated soils. Higher light-penetrated soils (LP70) showed significantly more fungal communities than the lower light-penetrated soils (LP40 and LP50) (p 4- N concentration in LP70 was significantly lower than those of LP40 and LP50, whereas the other chemical properties showed no significant difference among the soils. The cy19:0 to 18:1ω7c ratio was significantly lower in LP70 than in LP 40 and LP50 showing the negative correlation of light level with microbial stresses (p 4- N, actinomycetes, Gram-negative bacteria, and bacteria. Consequently, the amount of penetrating light was responsible for microbial compositions in the forest soils in correlation with the concentration of NH 4- N and soil temperature.

Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

Plants must constantly adjust their growth and defense responses to deal with the wide variety of stresses they encounter in their environment. Among phytohormones, brassinosteroids (BRs) are an important group of plant steroid hormones involved in numerous aspects of the plant lifecycle including growth, development and responses to various stresses including insect attacks. Here, we show that BRs regulate glucosinolate (GS) biosynthesis and function in insect herbivory. Preference tests and larval feeding experiments using the generalist herbivore, diamondback moth (Plutella xylostella), revealed that the larvae prefer to feed on Arabidopsis thaliana brassinosteroid insensitive 1 (bri1-5) plants over wild-type Ws-2 or BRI1-Flag (bri1-5 background) transgenic plants, which results in an increase in larval weight. Analysis of GS contents showed that 3-(methylsulfinyl) propyl GS (C3) levels were higher in bri1-5 than in Ws2 and BRI1-Flag transgenic plants, whereas sinigrin (2-propenylglucosinolate), glucoerucin (4-methylthiobutylglucosinolate) and glucobrassicin (indol-3-ylmethylglucosinolate) levels were lower in this mutant. We investigated the effect of brassinolide (BL) on GS biosynthesis in Arabidopsis and radish (Raphanus sativus L.) by monitoring the expression levels of GS biosynthetic genes, including MAM1, MAM3, BCAT4 and AOP2, which increased in a BL-dependent manner. These results suggest that BRs regulate GS profiles in higher plants, which function in defense responses against insects.

Kohlrabi (Brassica oleracea var. gongylodes L.) is an important vegetable of the Brassicaceae family. The main edible part of kohlrabi is the swollen stem. The purple cultivars make anthocyanin mainly in the peel of the swollen stem, while in the leaf, it is limited to the midrib, but green cultivars do not. Anthocyanins are advantageous for both plants as well as humans. Two anthocyanin compounds were detected by high pressure liquid chromatography (HPLC) only in the peel of the purple kohlrabi cultivar. Three MYBs, three bHLHs, and one WD40 TF were identified as the candidate regulatory genes in kohlrabi. There was an abundance of transcript levels for the late biosynthetic genes more specifically for BoF3′H, BoDFR, BoLDOX, and BoGST in the purple peel while scarcely detectable in other tissues for both cultivars. The expression of BoPAP2 and BoTT8 was higher in the peel of the purple cultivar than the green cultivar. The expression of BoMYBL2.2 orthologue of Arabidopsis MYBL2, a negative regulator of anthocyanins, was dramatically decreased in the purple peel. The expression of BoACO1, a key gene for ethylene biosynthesis, and BoNCED3, an important gene of the ABA pathway, was down- and upregulated, respectively, in the peel of purple kohlrabi.

Cultivated strawberry (Fragaria × ananassa Duch.) is a high value horticultural crop. In this study, the genetic diversity of 160 strawberry accessions was determined using five highly polymorphic simple sequence repeat (SSR) markers. Sixty different alleles were identified, with allele frequencies in the range of 0.006 to1. Similarity scores were in the range of 0.034 to 0.963 (average: 0.507). The accessions were categorized into five groups. Group 1 contained two diploid Fragaria vesca species and one unknown accession. Group 2 contained one accession (F x ananassa). Group 3 contained 20 F × ananassa accessions and six unknown accessions. Group 4 contained 48 F. × ananassa accessions, one octaploid Fragaria chiloensis species, and six unknown accessions while Group 5 contained 69 F. × ananassa accessions and six unknown accessions. Accessions within a pedigree were frequently grouped together. A total of 30 novel accessions were categorized alongside existing accessions. These results will allow breeders to develop strategies which incorporate more genetic diversity into new cultivars.

During cytokinesis in plants, trans-Golgi network-derived vesicles accumulate at the center of dividing cells and undergo various structural changes to give rise to the planar cell plate. However, how this conversion occurs at the molecular level remains elusive. In this study, we report that SH3 Domain-Containing Protein 2 (SH3P2) in Arabidopsis thaliana plays a crucial role in converting vesicles to the planar cell plate. SH3P2 RNAi plants showed cytokinesis-defective phenotypes and produced aggregations of vesicles at the leading edge of the cell plate. SH3P2 localized to the leading edge of the cell plate, particularly the constricted or curved regions of the cell plate. The BAR domain of SH3P2 induced tubulation of vesicles. SH3P2 formed a complex with dynamin-related protein 1A (DRP1A) and affected DRP1A accumulation to the cell plate. Based on these results, we propose that SH3P2 functions together with DRP1A to convert the fused vesicles to tubular structures during cytokinesis.

Rapid and reliable diagnosis of methicillin-resistant Staphylococcus aureus (MRSA) is crucial for guiding effective patient treatment and preventing the spread of MRSA infections. Nonetheless, further simplification of MRSA detection procedures to shorten detection time and reduce labor relative to that of conventional methods remains a challenge. Here, we have demonstrated a Clustered regularly interspaced palindromic repeats (CRISPR)-mediated DNA-FISH method for the simple, rapid and highly sensitive detection of MRSA; this method uses CRISPR associated protein 9/single-guide RNA (dCas9/sgRNA) complex as a targeting material and SYBR Green I (SG I) as a fluorescent probe. A dCas9/sgRNA-SG I based detection approach has advantages over monoclonal antibody in conventional immunoassay systems due to its ability to interact with the target gene in a sequence-specific manner. The detection limit of MRSA was as low as 10 cfu/ml and was found to be sufficient to effectively detect MRSA. Unlike conventional gene diagnosis methods in which PCR must be accompanied or genes are isolated and analyzed, the target gene can be detected within 30min with high sensitivity without performing a gene separation step by using cell lysates. We showed that the fluorescence signal of the MRSA cell lysate was more than 10-fold higher than that of methicillin-susceptible S. aureus (MSSA). Importantly, the present approach can be applied to any target other than MRSA by simply changing the single-guide RNA (sgRNA) sequence. Because dCas9/sgRNA-SG I based detection approach has proved to be easy, fast, sensitive, and cost-efficient, it can be applied directly at the point of care to detect various pathogens as well as MRSA in this study.

Objective: Asthma is characterized by airway hyperresponsiveness (AHR), inflammation, and obstruction. AHR to stimuli that indirectly cause bronchial smooth muscle (BSM) contractions via release of endogenous mediators is thought to better reflect airway inflammation than AHR to stimuli that act directly on BSM. Fractional exhaled nitric oxide (FeNO) is a useful parameter for noninvasive clinical airway inflammation assessments. Accordingly, this study aimed to examine the relationships of mannitol and methacholine challenge test outcomes with FeNO and the influence of inhaled corticosteroid treatment in children with asthma. Methods: One hundred thirty-four asthmatic children (89 males; ages: 5–17 years, median: 9 years) underwent spirometry, FeNO measurement, serum total/specific IgE testing, and blood eosinophil count. All subjects were challenged with mannitol dry powder (MDP; AridolH, Pharmaxis, Australia) and methacholine at 7-day intervals. Data of steroid-treated and steroid-naïve children were compared. Results: Positive responses to MDP and methacholine challenge tests were observed in 74.6% and 67.2% of total subject group, respectively, and 72 children had positive response to both challenge tests. The median FeNO level, response-dose ratio (RDR) of PC20 methacholine, and RDR of PD15 MDP were significantly higher in the steroid-treated group than in the steroid-naïve group (p < 0.001, 0.226, and 0.004, respectively). FeNO levels associated significantly with PD15 MDP and RDR PD15 MDP in total subject populations (p = 0.016 and 0.003, respectively); however, a significant correlation between FeNO and RDR PD15 MDP was observed only in the steroid-naïve group. Conclusions: Compared with AHR to methacholine, AHR to MDP more closely reflected the level of FeNO in steroid-naïve asthmatic children.

Motherhood as an identity is not inherently fixed but should be understood as an identity that changes, and grows as a result of relationships. However, a sterilized and sanitized image of motherhood is prevalent in the dominant culture. A heterosexual domesticated role of the mother is a barrier to appreciate the heterogeneous nature of motherhood. In this chapter, motherhood as an identity is explored as a self-giving and self-receiving relationships. This reciprocal relationship can generate life-giving power. This mundane and ordinary act can lead to a redemptive act that is scared and holy. Motherhood gives an alternative to the supposedly self-less salvific acts associated with masculine heroes. As a communal and everyday reality of motherhood illuminates God’s first desire which is to be in relationship.

Activated macrophages have a greater ability of phagocytosis against pathogens that is mediated by large-scale actin rearrangement. However, molecular machineries that conduct this task have not been fully identified. Here, we demonstrate an unanticipated role of TAGLN2, a 22-kDa actin-binding protein, in Toll-like receptor (TLR)-stimulated phagocytosis. TAGLN2 was greatly induced in macrophages in response to lipopolysaccharide (LPS), a ligand for TLR4, partly via the NF-κB pathway. TAGLN2-deficient macrophages (TAGLN2−/−) showed defective phagocytic functions of IgM- and IgG-coated sheep red blood cells as well as bacteria. Cell signaling pathways involved in actin rearrangement—PI3 kinase/AKT and Ras-ERK—were also down-regulated in LPS-stimulated TAGLN2-deficient macrophages. Moreover, TAGLN2−/− mice showed higher mortality after bacterial infection than wild-type littermates. Thus, our results revealed a novel function of TAGLN2 as a molecular armament required for host defense.

The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that senses environmental insults, was found to be involved in the pathology of psoriasis. Autophagy as an endogenous defense mechanism for environmental abnormalities was also reported to be related to pathogenesis of psoriasis. However, the cross-talk between AhR and autophagy and detailed mechanism of these in the pathogenesis of psoriasis are still mystery. The aim of this study was to explore the effects of AhR activation on the autophagy in HaCaT cells and to confirm the differences of expression of AhR and autophagy-related factors in human psoriasis lesions. HaCaT cells and M5 (IL-17A, IL-22, oncostatin M, TNF-u03b1, and IL-1u03b1)-stimulated HaCaT cells showing features of psoriasis were treated with the AhR agonists, TCDD and FICZ and AhR and CYP1A1 were detected by real-time RT-PCR and western blot. The levels of autophagy-related proteins were determined by western blot. We also measured the autophagosomes using immunostaining with an anti-LC3 antibody. In human psoriasis lesion, the degree of CYP1A1, AhR and LC3 expression was determined using immunohistochemistry, immunofluorescence and real time RT-PCR. FICZ and TCDD upregulated CYP1A1 and AhR in HaCaT cells. The expression of CYP1A1 and AhR were more increased in FICZ+M5, TCDD+M5-treated HaCaT cells. The LC3 punctuated cells were significantly decreased in HaCaT cells with FICZ and TCDD treatment. The production of autophgosomes was more suppressed in FICZ+M5, TCDD+M5-treated HaCaT cells. LC3, ATG5, p62, and beclin-1 were downregulated in TCDD and FICZ treated HaCaT cells. The expression of autophagy-related proteins were more decreased in FICZ+M5, TCDD+M5-treated HaCaT cells. In human psoriasis lesion, the expression of CYP1A1 and AhR was increased and LC3 was decreased compared to normal skin. These data demonstrated that AhR could modulate the autophagy in human HaCaT cells, which suggest a new mechanism in pathogenesis of psoriasis.

The bacterial pathogen Pantoea sp. strain PPE7 causes severe soft rot disease in the king oyster mushroom, Pleurotus eryngii. Typical symptoms of the disease include a dark-brown water drop at early stages of infection, ultimately leading to soft rot accompanied with an offensive odour. In this study, we showed that inoculation of Flammulina velutipes and Agaricus bisporus mushrooms with Pantoea sp. strain PPE7 also resulted in severe disease, including water-soaked lesions and soft rot symptoms. Treatment with chlorine solution (175 to 700 ppm active chlorine) significantly reduced the incidence of soft rot symptoms in Pleurotus eryngii. However, exposure to concentrations greater than 350 ppm caused harmful effects such as bleaching on the cap of the fruit body. Calcium hypochlorite solutions with 175 ppm active chlorine were effective for reducing soft rot disease of Pleurotus eryngii without affecting the mushroom yield.

In this study, we investigated anthocyanin accumulation and gene expression in response to light and dark conditions during the development of white and red cabbage seedlings. Two-day-old white cabbage seedlings expressed the highest transcript level for most of the genes under light conditions. Red cabbage also showed higher expression under light than under dark conditions, although gene expression (evaluated based on transcript levels normalized to that of a housekeeping gene) in 2-day-old red cabbage sprouts was much lower than that in the corresponding white cabbage sprouts. Trends in anthocyanin accumulation were similar for red and white cabbage but much greater accumulation was observed in red cabbage. Anthocyanin levels were higher in seedlings grown under light conditions compared to those grown under dark conditions for both cabbage cultivars. Especially, red cabbage accumulated 1.94–4.05 times greater total anthocyanin in 4-, 6-, 8-, and 10-day-old seedlings, when compared to white cabbage cultivar under light/dark and dark conditions. Our findings can improve understanding of the effects of light on accumulation of secondary metabolites in the seedling stages of various crops.

In this paper, an experimental investigation was carried out to evaluate the application of engineered cement composite (ECC) wing wall elements for seismic strengthening of reinforced concrete (RC) buildings with non-ductile reinforcement details. The ECC is mixed with cement, silica fume, fly ash and polyvinyl alcohol (PVA) fiber. The ECC wing wall elements were fitted on three side of RC frame; upper beam, lower beam and column. Two specimens, non-ductile RC bare frame and RC frame strengthened with ECC wing wall, were made for this study. These specimens were made as a third scale for this study. The specimens were loaded through 1,000 kN actuator that consists of two cycles at a displacement level. The test results showed that the seismic strengthening method of ECC wing wall elements had a significant effect to improve seismic performance of non-ductile RC bare frame. As a result, the application of ECC wing wall elements in non-ductile RC bare frame may be effective alternative to seismic retrofit of non-ductile buildings.

The present study investigated variations in soil microbial communities by fatty acid methyl ester (FAME) and the chemical properties at 24 sites of upland soils in Gyeongnam Province. The electrical conductivity of the soil under potato cultivation was significantly higher than those of the red pepper and soybean soils (p < 0.05). The gram-negative bacteria community in potato soil was significantly lower than those in the garlic and soybean soils (p < 0.05). The communities of actinomycetes and arbuscular mycorrhizal fungi in the red pepper soil were significantly higher than those in the potato soil (p < 0.05). In addition, the cy17:0 to 16:1ω7c ratio was significantly lower in red pepper, soybean, and garlic soils compared with potato soil, indicating that microbial stress decreased. Consequently, differences in soil microbial community were highly associated with cultivated crop species, and this might be resulted from the difference in soil chemical properties.

Brassica rapa and Raphanus sativus are strictly self-incompatible (SI) plants; however, xBrassicoraphanus (baemoochae) is an intergeneric hybrid synthesized following hybridization of B. rapa and R. sativus that is self-compatible (SC). Self-incompatibility in Brassicaceae is controlled by multiple alleles of the S-locus. Two S-locus genes, SRK (S-locus receptor kinase) and SP11/SCR (S-locus protein 11 or S-locus cysteine-rich), have been reported to date, both of which are classified into class I and II. In this study, we investigate if there is an alteration in the structure or the expression in SRK or SP11 genes behind the alteration of SI to SC in baemoochae. Class I and II SRK were identified by PCR of the genomic DNA of baemoochae using SRK-specific universal primers. Cloning and sequencing of both classes of SRK was conducted and compared with SRK genes of parental species. Comparison analysis showed no genomic alterations and both of them showed expression in the stigma. Similarly, SP11 genes also showed no genomic alterations and normally are expressed in the anther. Other SI-related genes (MLPK, ARC1, THL) also showed normal expression in the stigma and anther. Taken together, these results revealed that no structural/gene expression change in these genes was responsible for the breakdown of SI in baemoochae. Rather, the transformation from SI parents to SC descendants could be responding to epigenetic mechanisms.

In 2009-2010, unusual symptoms were observed on Pleurotus eryngii grown in mushroom farms in Gyeongnam Province, Republic of Korea. One of the main symptoms was a cobweb-like growth of fungal mycelia over the surface of the mushroom. The colonies on the surface rapidly overwhelmed the mushrooms and developed several spores within 3-4 days. The colonized surface turned pale brown or yellow. The fruit body eventually turned dark brown and became rancid. Koch's postulates were completed by spraying and spotting using isolated strains. The phylogenetic tree obtained from the internal transcribed spacer sequence analysis showed that the isolated fungal pathogen corresponded to Cladobotryum mycophilum (99.5%). In the fungicide sensitivity tests, the ED50 values for the isolate with respect to benomyl and carbendazim were from 0.29 to 0.31 ppm. Benzimidazole fungicides were most effective against C. mycophilum, a causal agent of cobweb disease in P. eryngii.

In every city, low-volume roads and parking lots are composed of dense-graded pavements that increase storm-water runoff; however, urban flooding problems can be prevented if an appropriate pavement system is achieved. One preferable solution to this problem for low volume roads and parking lots is to use permeable interlocking block pavement. To determine the storage capacity of this kind of pavement, a series of material property tests were performed such as specific gravity, void ratios, and porosity etc., which was further used to calculate the water storage capacity of the pavement. And rainfall simulation tests were performed to evaluate surface runoff, delay time, storage capacity, and outflows over two hours. A regression analysis showed that the water storage capacity measurement from rain simulation tests agreed with the storage capacity calculation depending on laboratory materials-properties tests, based on their high correlation and coefficient values, each within the 95% confidence interval.

AtRGP (AT4G17080, Arabidopsis thaliana reduction in growth and productivity) contains two N-terminal transmembrane helices and seven membrane occupation and recognition nexus motifs at its C-terminus, and associates with phosphatidylinositol phosphate kinase. To elucidate the function of AtRGP, we employed mutant plants to analyze gene expression, plant phenotypes, protein localization, structure and function of the chloroplast, and freezing tolerance. Overexpression of AtRGP increased growth rate, hypocotyl elongation, leaf size, seed production, photosynthetic rate, and freezing tolerance, and promoted chloroplast organization and stacking of grana. By contrast, Atrgp null mutants exhibited a smaller plant size, reduced seed production, photosynthetic rate, and freezing tolerance, and displayed abnormal chloroplast organization with insufficient stacking of grana. Considering these data, we postulate that AtRGP may bind transiently to the chloroplast envelope and interact with other proteins under certain conditions, thereby regulating cellular processes involved in growth and abiotic stress responses.

This paper reports the experimental results to evaluate in-plane shear performance of insulated concrete sandwich panel (ICSP) with glass fiber-reinforced polymer (GFRP) grid shear connectors. The variables considered in this study are the grid size (35 and 53mm) of GFRP shear connectors and the types of insulation (expanded polystyrene, EPS and extruded polystyrene with special slots, XPSS). For loading in-plane shear force to interface between inner and outer wall of ICSP system, the ICSP specimens were supported vertically at the bottom edge of the two concrete outer walls by steel blocks. The test results indicate that ICSP with XPSS developed higher shear flow strengths in ICSP with EPS when 35mm spacing of GFRP grid is used. Also, the test results indicated that as the grid spacing of GFRP shear connector decreases, the shear flow strength of ICSP with XPSS insulation was higher, but the shear flow strength of ICSP with EPS insulation was lower.

In this paper, an experimental investigation was carried out to examine the influence of hooked end steel fiber volume fraction and curing conditions on the compressive performance of concrete produced by using ordinary portland cement (OPC) and alkali-activated slag (AAS). Three different volume fractions of 0.5%, 1.0% and 1.5% were used in OPC and AAS concrete mixtures. Cylindrical specimens with 100 x 200mm were tested for compressive behavior of both concretes at 3, 7 and 28 days of curing age. Test results showed that curing conditions had a significant effect on compressive properties in the hardened OPC and AAS concretes. The addition of steel fibers generated a decrease in compressive strength of OPC while an increase in the compressive strength of AAS concrete was shown with adding steel fiber.

QTLs and candidate gene markers associated with leaf morphological and color traits were identified in two immortalized populations of Brassica rapa, which will provide genetic information for marker-assisted breeding. Brassica rapa is an important leafy vegetable consumed worldwide and morphology is a key character for its breeding. To enhance genetic control, quantitative trait loci (QTLs) for leaf color and plant architecture were identified using two immortalized populations with replications of 2 and 4 years. Overall, 158 and 80 QTLs associated with 23 and 14 traits were detected in the DH and RIL populations, respectively. Among them, 23 common robust-QTLs belonging to 12 traits were detected in common loci over the replications. Through comparative analysis, five crucifer genetic blocks corresponding to morphology trait (R, JU, F and E) and color trait (F, E) were identified in three major linkage groups (A2, A3 and A7). These might be key conserved genomic regions involved with the respective traits. Through synteny analysis with Arabidopsis, 64 candidate genes involved in chlorophyll biosynthesis, cell proliferation and elongation were co-localized within QTL intervals. Among them, SCO3, ABI3, FLU, HCF153, HEMB1, CAB3 were mapped within QTLs for leaf color; and CYCD3;1, CYCB2;4, AN3, ULT1 and ANT were co-localized in QTL regions for leaf size. These robust QTLs and their candidate genes provide useful information for further research into leaf architecture with crop breeding.

There is an increasing awareness that as a result of structural variation, a reference sequence representing a genome of a single individual is unable to capture all of the gene repertoire found in the species. A large number of genes affected by presence/absence and copy number variation suggest that it may contribute to phenotypic and agronomic trait diversity. Here we show by analysis of the Brassica oleracea pangenome that nearly 20% of genes are affected by presence/absence variation. Several genes displaying presence/absence variation are annotated with functions related to major agronomic traits, including disease resistance, flowering time, glucosinolate metabolism and vitamin biosynthesis., Brassica oleracea is a single species that includes diverse crops such as cabbage, broccoli and Brussels sprouts. Here, the authors identify genes not captured in existing B. oleracea reference genomes by the assembly of a pangenome and show variations in gene content that may be related to important agronomic traits