Search

Objective Ferroptosis plays an important role in traumatic brain injury (TBI). The p53 protein is a major mediator of ferroptosis. However, the role of p53-mediated ferroptosis in TBI has not been studied. Sirtuin 2 (SIRT2) exerts a protective effects role in TBI, although the underlying mechanism of this protection remains unclear. In the present study, we tested the hypothesis that that SIRT2 mitigates TBI by regulating p53-mediated ferroptosis. Methods and results To model TBI in mice, we used the controlled cortical impact (CCI) injury method. We found that ferroptosis was significantly activated by CCI, and peaked 3 days following CCI, as evidenced by upregulation of GPX4 and SLC7A11, increased content of decreases glutathione, lipid peroxidation, malondialdehyde and ferrous ion. Inhibition of ferroptosis significantly alleviated neurological indications and brain edema. In addition, knockout of p53 significantly blocked ferroptosis following CCI. Furthermore, we found that inhibition of SIRT2 upregulated the acetylation of p53, as well as p53 expression, and exacerbated ferroptosis following CCI. Interestingly, knockout of p53 rescued the SIRT2 inhibition-induced exacerbation of ferroptosis. Conclusions These findings indicate that p53-mediated ferroptosis contributes to the pathogenesis of TBI. Furthermore, we demonstrate that SIRT2 exerts a neuroprotective effect against TBI by suppressing p53-mediated ferroptosis.

Nelumbo nucifera, more commonly known as the Indian lotus, is an important plant that has been incorporated into traditional herbal remedies along the years. Even today, lotus leaves are considered reservoirs for bioactive compounds that can be used as nutritional supplements to treat various human diseases. However, despite the wide ranging biological activities of lotus polysaccharides, limited information is available regarding the anti-osteoporotic effects of these substances. The aim of this study was to investigate the beneficial effects of pectinase-assisted extractable polysaccharides from lotus leaves (LLEP) on estrogen deficiency-induced bone loss and osteoclast differentiation in bone marrow-derived macrophages. We found that LLEP markedly inhibited receptor activator of the nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation in a dose-dependent manner. It also revoked RANKL-induced activation of osteoclastogenic signals such as the expression of key transcription factors (i.e., c-Fos and nuclear factor of activated T cells cytoplasmic 1), resulting in a decrement in osteoclast-specific marker gene expressions. Microcomputed tomography and morphometric analysis revealed that a four-week oral administration of LLEP notably decreased trabecular bone loss. Taken together, our results suggest that LLEP can mitigate estrogen deficiency-induced bone loss by suppressing osteoclastogenesis, which makes it an excellent candidate for combating osteoporosis.

This study aimed to determine the probiotic potential of Pediococcus acidilactici M76 (PA-M76) for lactic acid fermentation of black raspberry extract (BRE). PA-M76 showed outstanding probiotic properties with high tolerance in acidic GIT environments, broad antimicrobial activity, and high adhesion capability in the intestinal tract of Caenorhabditis elegans. PA-M76 treatment resulted in significant increases of pro-inflammatory cytokine mRNA expression in macrophages, indicating that PA-M76 elicits an effective immune response. When PA-M76 was used for lactic acid fermentation of BRE, an EPS yield of 1.62 g/L was obtained under optimal conditions. Lactic acid fermentation of BRE by PA-M76 did not significantly affect the total anthocyanin and flavonoid content, except for a significant increase in total polyphenol content compared to non-fermented BRE (NfBRE). However, fBRE exhibited increased DPPH radical scavenging activity, linoleic acid peroxidation inhibition rate, and ABTS scavenging activity of fBRE compared to NfBRE. Among the 28 compounds identified in the GC-MS analysis, esters were present as the major groups. The total concentration of volatile compounds was higher in fBRE&nbsp, than that in NfBRE. However, the undesirable flavor of terpenes decreased. PA-M76 might be useful for preparing functionally enhanced fermented beverages with a higher antioxidant activity of EPS and enhanced flavors.

SUMMARY Myeloid lineage cells use TLRs to recognize and respond to diverse microbial ligands. Although unique transcription factors dictate the outcome of specific TLR signaling, whether lineage-specific differences exist to further modulate the quality of TLR-induced inflammation remains unclear. Comprehensive analysis of global gene transcription in human monocytes, monocyte-derived macrophages, and monocyte-derived dendritic cells stimulated with various TLR ligands identifies multiple lineage-specific, TLR-responsive gene programs. Monocytes are hyperresponsive to TLR7/8 stimulation that correlates with the higher expression of the receptors. While macrophages and monocytes express similar levels of TLR4, macrophages, but not monocytes, upregulate interferon-stimulated genes (ISGs) in response to TLR4 stimulation. We find that TLR4 signaling in macrophages uniquely engages transcription factor IRF1, which facilitates the opening of ISG loci for transcription. This study provides a critical mechanistic basis for lineage-specific TLR responses and uncovers IRF1 as a master regulator for the ISG transcriptional program in human macrophages., Graphical abstract, In brief Song et al. examine the responses of human myeloid populations to bacterial and viral ligands and discover the lineage-specific induction of proinflammatory gene expression. TLR4 signaling in macrophages, but not monocytes, enables nuclear translocation of IRF1, which acts as a pioneer transcription factor to increase the chromatin accessibility of IFNB1 and ISG loci.

Significance Inflammation has largely been studied in the context of protein-coding genes. Recent studies have uncovered lncRNAs as important regulators of immunity. The functional characterization of these genes remains an active area of research. In this study, we identify GAPLINC as a functionally conserved lncRNA between human and mouse. GAPLINC depletion results in enhanced expression of immune response genes that are direct NF-κB targets. Astoundingly, we observe that Gaplinc knockout mice show resistance to LPS-induced endotoxic shock and find that basal expression of inflammatory genes prevents clot formation to protect against multiorgan failure and death. These findings have implications in the treatment of sepsis, in which new therapies targeting lncRNAs can contribute valuable information in understanding inflammation and improving patient outcome., Recent studies have identified thousands of long noncoding RNAs (lncRNAs) in mammalian genomes that regulate gene expression in different biological processes. Although lncRNAs have been identified in a variety of immune cells and implicated in immune response, the biological function and mechanism of the majority remain unexplored, especially in sepsis. Here, we identify a role for a lncRNA—gastric adenocarcinoma predictive long intergenic noncoding RNA (GAPLINC)—previously characterized for its role in cancer, now in the context of innate immunity, macrophages, and LPS-induced endotoxic shock. Transcriptome analysis of macrophages from humans and mice reveals that GAPLINC is a conserved lncRNA that is highly expressed following macrophage differentiation. Upon inflammatory activation, GAPLINC is rapidly down-regulated. Macrophages depleted of GAPLINC display enhanced expression of inflammatory genes at baseline, while overexpression of GAPLINC suppresses this response. Consistent with GAPLINC-depleted cells, Gaplinc knockout mice display enhanced basal levels of inflammatory genes and show resistance to LPS-induced endotoxic shock. Mechanistically, survival is linked to increased levels of nuclear NF-κB in Gaplinc knockout mice that drives basal expression of target genes typically only activated following inflammatory stimulation. We show that this activation of immune response genes prior to LPS challenge leads to decreased blood clot formation, which protects Gaplinc knockout mice from multiorgan failure and death. Together, our results identify a previously unknown function for GAPLINC as a negative regulator of inflammation and uncover a key role for this lncRNA in modulating endotoxic shock.

Background Although anxiety disorders are one of the most common mental illness in population, antianxiety drugs often only have single action targets, require long-term use, and are associated with many adverse reactions and dependencies. Professor Yan Zhaojun from Shandong Provincial Hospital of Traditional Chinese Medicine (TCM) has applied the modified Renshu Powder, a TCM formula, to treat anxiety disorders, with satisfactory outcomes. Here, we investigated the mechanism of action of two core herbs (prepared Rehmannia root and Chinese arborvitae kernel) in the Renshu Powder in the treatment of anxiety disorders by using network pharmacology approaches. Methods Candidate compounds of the herb pair of prepared Rehmannia root-Chinese arborvitae kernel were extracted via the Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform. The targets of action of the main compounds were collected using the SwissTargetPrediction database. Targets associated with anxiety disorders were retrieved from DisGeNET, Online Mendelian Inheritance in Man (OMIM), DrugBank, GeneCards, and Comparative Toxicogenomics Database (CTD) databases. The compound-target interaction network was constructed by Cytoscape 3.7.2 software, and the protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) platform. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses the data by using Metascape. Results The main active compounds of the herb pair included arachidonic acid, stigmasterol, and beta-sitosterol. The key targets included Nitric Oxide Synthase 3 (NOS3), Epidermal growth factor (EGF), Prostaglandin-Endoperoxide Synthase 2 (PTGS2), Caspase 3 (CASP3), Mitogen-Activated Protein Kinase 1 (MAPK1), Peroxisome proliferator-activated receptor gamma (PPARG), RELA Proto-Oncogene, NF-KB Subunit (RELA), Estrogen Receptor 1 (ESR1), Solute Carrier Family 6 Member 4 (SLC6A4), and Phosphatase and Tensin homolog deleted on chromosome 10 (PTEN). Anxiety disorder-related GO analysis mainly involved synaptic signaling, neurotransmitter receptor activity, and G protein-coupled neurotransmitter receptor activity. The KEGG pathways involved neuroactive ligand-receptor interaction, serotonergic synapse, PI3K/AKT/mTOR signaling pathway, and MAPK signaling pathway. Conclusions The mechanism of action of the prepared Rehmannia root-Chinese arborvitae kernel in treating anxiety disorders involves multiple ingredients, multiple targets, and pathways.

BackgroundXylitol accumulation is a major barrier for efficient ethanol production through heterologous xylose reductase-xylitol dehydrogenase (XR-XDH) pathway in recombinantSaccharomyces cerevisiae. Mutated NADH-preferring XR is usually employed to alleviate xylitol accumulation. However, it remains unclear how mutated XR affects the metabolic network for xylose metabolism. In this study, haploid and diploid strains were employed to investigate the transcriptional responses to changes in cofactor preference of XR through RNA-seq analysis during xylose fermentation.ResultsFor the haploid strains, genes involved in xylose-assimilation (XYL1,XYL2,XKS1), glycolysis, and alcohol fermentation had higher transcript levels in response to mutated XR, which was consistent with the improved xylose consumption rate and ethanol yield. For the diploid strains, genes related to protein biosynthesis were upregulated while genes involved in glyoxylate shunt were downregulated in response to mutated XR, which might contribute to the improved yields of biomass and ethanol. When comparing the diploids with the haploids, genes involved in glycolysis and MAPK signaling pathway were significantly downregulated, while oxidative stress related transcription factors (TFs) were significantly upregulated, irrespective of the cofactor preference of XR.ConclusionsOur results not only revealed the differences in transcriptional responses of the diploid and haploid strains to mutated XR, but also provided underlying basis for better understanding the differences in xylose metabolism between the diploid and haploid strains.

Determining the layers of gene regulation within the innate immune response is critical to our understanding of the cellular responses to infection and dysregulation in disease. We identified a conserved mechanism of gene regulation in human and mouse via changes in alternative first exon (AFE) usage following inflammation, resulting in changes to isoform usage. Of these AFE events, we identified 50 unannotated transcription start sites (TSS) in mice using Oxford Nanopore native RNA sequencing, one of which is the cytosolic receptor for dsDNA and known inflammatory inducible gene, Aim2. We show that this unannotated AFE isoform of Aim2 is the predominant isoform transcribed during inflammation and contains an iron-responsive element in its 5′UTR enabling mRNA translation to be regulated by iron levels. This work highlights the importance of examining alternative isoform changes and translational regulation in the innate immune response and uncovers novel regulatory mechanisms of Aim2.Summary SentenceAlternative first exon usage was the major splicing event observed in macrophages during inflammation, which resulted in the elucidation of a novel isoform and iron mediated regulatory mechanism of the protein coding gene, Aim2.

Ovarian cancer (OC) is one of gynecological malignancies that seriously affects women's health. Mounting evidence demonstrated that long noncoding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) play important roles in various biological processes related to the pathogenesis of OC. This research aimed to investigate the regulatory mechanism of lncRNA SCAMP1/miR-137/CXCL12 (C-X-C motif chemokine ligand 12) axis on OC progression. In this study, we found that SCAMP1 was highly expressed in OC cells, which promoted OC cell invasion and angiogenesis. In addition, our research confirmed that SCAMP1 could bind with miR-137, and SCAMP1 sponged miR-137 to accelerate the progression of OC. We also observed that CXCL12 was a downstream target gene for miR-137, and miR-137 targeted CXCL12 to participate in the regulation of OC. Finally, through TCGA database, we found that SCAMP1 (or CXCL12) was upregulated as well as miR-137 was downregulated in OC tissues, and high (or low) level of them was associated with poor prognosis. miR-137 expression was negatively correlated with SCAMP1 (or CXCL12) expression, and SCAMP1 expression was positively correlated with CXCL12 expression in OC. In summary, our study clarified the role of SCAMP1/miR-137/CXCL12 axis in OC, and this finding may provide a potential therapeutic target of OC.

We present the data showing that overexpression of the Arabidopsis zinc finger homeodomain protein ZHD5 promotes shoot regeneration and confers other cytokinin-associated phenotypes. To isolate protein factors that bind to the promoter element of ABF2, which is a key regulator of ABA responsive gene expression, we carried out yeast-one-hybrid screen. Four zinc finger homeodomain (ZF-HD) protein genes were identified from the screen. Expression study showed that one of the ZF-HD proteins, ZHD5, is induced by various cytokinins (CKs). We generated and analyzed ZHD5 overexpression (OX) lines to study its function. Our results revealed that ZHD5 promoted shoot regeneration efficiency significantly, obviating CK requirement. Other CK-related phenotypes, such as enhanced cell proliferation and pathogen resistance, were also observed. At the molecular level, changes in the expression levels of a number of genes relevant to the phenotypes were observed. Collectively, our results provde strong evidence that ZHD5 is involved in shoot regeneration and other aspects of CK response.

HPV16 is the predominant type of HPV causing invasive cervical cancer. However, the underlying molecular mechanism of the unparalleled carcinogenic power of HPV16 compared to other types of high-risk (HR)-HPV including HPV18 remains elusive. The PDZ binding motif (PBM) of high-risk HPV E6 plays an important role in neoplasia and progression of cervical cancer. HPV16 E6 rather than HPV18 E6, interacted with NHERF1 by its PBM region, and induced degradation of NHERF1. NHERF1 retarded the assembly of cytoskeleton by downregulation of ACTN4, thereby inhibited the migration and invasion of cervical cancer cells in both cell and mouse model. HPV16 E6 was confirmed to enhance actin polymerization with increased ACTN4 level by downregulation of NHERF1, and result in enhanced migration and invasion of cervical cancer cells. GSEA analysis of cervical cancer specimens also showed that HPV16 E6 rather than HPV18 E6, was significantly associated with actin cytoskeleton assembly. That downregulation of NHERF1 by HPV16 E6 promoted cytoskeleton assembly and cell invasion, was an important cause in cervical cancer carcinogenesis. These findings provided the differential mechanism between HPV16 E6 and HPV18 E6 in the development and progression of cervical cancer, which may partially explain the differences of carcinogenic power between these two types of HR-HPVs.

In this study, enzyme-assisted extraction was used to isolate functional polysaccharides from Korean ginseng (Panax ginseng Meyer) and the physicochemical and biological properties of the extracted polysaccharides were investigated, comparing with those from traditional hot-water extraction (FGWP). In macrophages, their effects on cytokines production could be ordered as FGEP-CA ≥ FGEP-A > FGEP-C > FGWP, suggesting that FGEP-CA (combined cellulase- and α-amylase-extracted polysaccharide) is a potent immunostimulator. In addition, enzymatic digestion led to differences in the monosaccharide profile of the extract. FGWP mainly consisted of rhamnose, arabinose, galactose, galacturonic acid, and glucose in molar percentages of 1.8:10.1:9.2:17.8:60.6, whereas FGEP-CA was 3.2:11.4:16.5:22.3:45.8, respectively, suggesting that enzyme-assisted extraction of ginseng polysaccharides produces a higher proportion of pectin polysaccharides. The HPLC profile of FGEP-CA also showed lower and more heterogeneous molecular weights than FGWP did. In cyclophosphamide-induced immunosuppressed mice, FGEP-CA administration ameliorated decreased spleen and thymus indices (200 mg/kg), lymphocyte proliferation, natural killer cell activity, leukocyte counts, and the serum cytokines, interleukin-2, interleukin-6, and interferon-γ (100 and 200 mg/kg). These results suggest that enzyme-assisted extraction using cellulase and α-amylase is an effective method for the preparation of functional polysaccharides from fresh Korean ginseng, and FGEP-CA could be utilized as a potential immune-stimulatory agent.

Highlights • We aimed to identify the optimum mixing ratio for various ingredients. • The extract yield and amounts of NO and IL-6 were set to maximum for modeling. • At the optimum values, and 35.3% yield, 14.7% NO, and 6.6% IL-6 were obtained. • The combined extract increased the spleen index and T- and B-cell proliferation. • These results provide a basis for the development of functional extracts and drinks., We aimed to identify the optimum mixing ratio for various ingredients to obtain complex extracts with high extract yields and immune-enhancing activity in animals. The extract yield and amounts of nitric oxide (NO) and interleukin (IL)-6 were set to maximum for modeling predictions. The predicted optimum values for the mixing ratio were 49.5% for Rhodiola crenulata, 26.1% for Astragalus membranaceus, and 24.4% for Panax quinquefolius, and the predicted response values were 31.5% yield, 13.4% NO production, and 6.1% IL-6 production; actual values were 35.3% yield, 14.7% NO, and 6.6% IL-6. The optimum mixing ratio extract (OMRE) was used for the animal experiments. Treating mice with OMRE at 200 mg/kg produced significant increases in spleen indexes and T-cell/B-cell proliferation. OMRE treatment increased IL-10 and IL-6 production in concanavalin A- and lipopolysaccharide-induced T- and B- lymphocytes, respectively. These results provide a basis for the development of functional extracts and drinks.

This study compared and examined the effects of aroma gargling, cold water gargling, and wet gauze application on thirst, halitosis, and sore throat in patients after spine surgery. A quasiexperimental pretest/posttest control group design was employed. Samples were total 70 patients (aroma gargling: 24 samples, cold gargling: 24 samples, and wet gauze: 22 samples) after spine surgery in K Hospital in Seoul, Korea. The aroma gargle solution as an experimental intervention was prepared by blending peppermint, tea tree, and lemon oils at a ratio of 1:2:2. A 60 cc of aroma gargle solution was used 3 times for 15 to 20 seconds. The visual analog scale was used to measure the degrees of thirst and sore throat, and a portable device was used to examine the degree of halitosis. There were significant differences in the degrees of thirst, halitosis, and sore throat according to interaction between group and duration. In the comparison among 3 groups, aroma gargling provided better oral health by decreasing thirst, halitosis, and sore throat for patients with spine surgery. Aroma gargling can be utilized as an effective nursing intervention for decreasing thirst, halitosis, and sore throat for patients with spine surgery in clinical practice. Results suggest, therefore, that health professionals should consider an array of methods including aroma gargling for patients after spine surgery.

A gene encoding cinnamoyl esterase (CE), which breaks down chlorogenic acid (ChA) into caffeic and quinic acids, was cloned from Lactobacillus helveticus KCCM 11223. The gene with an open reading frame of 759 nucleotides was expressed in Escherichia coli, which resulted in a 51.6-fold increase in specific activity compared to L. helveticus KCCM 11223. The recombinant CE exists as a monomeric enzyme having a molecular weight of 27.4 kDa. Although the highest activity was observed at pH 7, the enzyme showed stable activity at pH 4.0–10.0. Its optimum temperature was 65°C, and it also possessed a thermophilic activity: the half-life of CE was 24.4 min at 65°C. The half-life of CE was 145.5, 80.5, and 24.4 min at 60, 62, and 65°C, respectively. The Km and Vmax values for ChA were 0.153 mM and 559.6 µM/min, respectively. Moreover, the CE showed the highest substrate specificity with methyl caffeate among other methyl esters of hydroxycinnamic acids such as methyl ferulate, methyl sinapinate, methyl p-c...

BackgroundSystemic lupus erythematosus (SLE) is a clinically heterogeneous autoimmune disease characterized by the development of anti-nuclear antibodies. Susceptibility to SLE is multifactorial, with a combination of genetic and environmental risk factors contributing to disease development. Like other polygenic diseases, a significant proportion of estimated SLE heritability is not accounted for by common disease alleles analyzed by SNP array-based GWASs. Death-associated protein 1 (DAP1) was implicated as a candidate gene in a previous familial linkage study of SLE and rheumatoid arthritis, but the association has not been explored further.ResultsWe perform deep sequencing across the DAP1 genomic segment in 2032 SLE patients, and healthy controls, and discover a low-frequency functional haplotype strongly associated with SLE risk in multiple ethnicities. We find multiple cis-eQTLs embedded in a risk haplotype that progressively downregulates DAP1 transcription in immune cells. Decreased DAP1 transcription results in reduced DAP1 protein in peripheral blood mononuclear cells, monocytes, and lymphoblastoid cell lines, leading to enhanced autophagic flux in immune cells expressing the DAP1 risk haplotype. Patients with DAP1 risk allele exhibit significantly higher autoantibody titers and altered expression of the immune system, autophagy, and apoptosis pathway transcripts, indicating that the DAP1 risk allele mediates enhanced autophagy, leading to the survival of autoreactive lymphocytes and increased autoantibody.ConclusionsWe demonstrate how targeted sequencing captures low-frequency functional risk alleles that are missed by SNP array-based studies. SLE patients with the DAP1 genotype have distinct autoantibody and transcription profiles, supporting the dissection of SLE heterogeneity by genetic analysis.

Botanical polysaccharides have been widely known to possess immunological activity. The objective of this study was to investigate the molecular mechanisms underlying the immunostimulatory properties of polysaccharides isolated from barley leaf (Hordeum vulgare L.) (BLE0) in splenocytes and cyclophosphamide (CYP)-induced immunosuppressed mice. BLE0 showed cell proliferative activity and markedly increased the secretion of both Th1-cytokines (IFN-γ and IL-2) and Th2-cytokines (IL-4 and IL-10) in CD3/CD28-activated splenocytes. Molecular data revealed that BLE0 up-regulated the expression of T-bet with enhanced phosphorylation of Janus kinase (JAK)-signal transducer and activator of transcription (STAT) 1 signaling pathway. BLE0 also increase the phosphorylation of GATA3 via toll-like receptor (TLR) 2-mediated signaling pathway with nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein 1 (AP-1) activation. Oral administration of BLE0 effectively improved CYP-induced decrease of body weight, splenocyte proliferation, and natural killer (NK) cell cytotoxic activity and significantly increased Th1 and Th2 cytokines, T-bet, and GATA3 mRNA expression. Dietary intake of BLE0 improves the immunological manifestations by stimulating both Th1 and Th2 responses via JAK/STAT1/T-bet and TLR2/GATA3, respectively.

The pubertal period is an important window during the development of the female reproductive system. Development of the pubertal ovary, which supplies the oocytes intended for fertilization, requires growth factors, hormones, and neuronal factors. It has been reported that zinc oxide nanoparticles (ZnO NPs) cause cytotoxicity of neuron cells. However, there have been no reports of the effects of ZnO NPs on neuronal factors and neuroendocrine cells in the ovary (in vivo). For the first time, this in vivo study investigated the effects of ZnO NPs on gene and protein expression of neuronal factors and the population of neuroendocrine cells in ovaries. Intact NPs were detected in ovarian tissue and although ZnO NPs did not alter body weight, they reduced the ovary organ index. Compared to the control or ZnSO4 treatments, ZnO NPs treatments differentially regulated neuronal factor protein and gene expression, and the population of neuroendocrine cells. ZnO NPs changed the contents of essential elements in the ovary; however, they did not alter levels of the steroid hormones estrogen and progesterone. These data together suggest that intact ZnO NPs might pose a toxic effect on neuron development in the ovary and eventually negatively affect ovarian developmental at puberty.

The accumulation of free polyamines (PAs) plays critical role in plants under abiotic stress conditions, while protective traits of bound PAs receiving much less attention, our understanding of the regulation of free PAs converted to bound PAs is limited. In this study, we demonstrated that the PA mediator transglutaminase (TGase) play an important role in improving tomato tolerance to salinity stress, in part because its critical role in inducing accumulation of bound PAs under salt stress. TGase expression was induced upon salt stress. Exogenous phenanthroline (O-phen, a TGase and bound PAs synthesis inhibitor) abolished the TGase-mediated salt tolerance with terminating bound PAs accumulation in TGaseOE plants. O-phen also had a negatively altered on Na+/K+ uptake or homeostasis under salt stress in leaves of TGaseOE plants. Instead, the overexpression of TGase effectively alleviated salt-induced oxidative damage and reduced reactive oxygen species (ROS) over-production, but it was significantly aggravated by O-phen in the TGaseOE plants. Although the ROS generating enzymes (PA oxidase, diamine oxidase and NADPH oxidase) was not affected by application of O-phen, the antioxidant enzyme (superoxide dismutase, catalase, ascorbate peroxidase and monodehydroascorbate reductase) activity was significantly decreased by exogenous O-phen in plants. Furthermore, further analysis showed that the antioxidant enzyme activity positively and closely correlated with the level of bound PAs in TGaseOE plant. Together, these results uncovered a probably function mechanism of TGase-mediated antioxidant enzyme activity to reduced salt-induced oxidative damage, which is partially depended on bound PAs accumulation.

Polysaccharides isolated from various plants are considered precious bioactive materials owing to their potent biological activities. Previously, we prepared a polysaccharide fraction (BLE0) isolated from young barley leaves (Hordeum vulgare L.), demonstrating its anti-osteoporotic and immunostimulatory activities. However, data regarding BLE0 toxicity is lacking. To establish its safety, in vitro genotoxicity (chromosomal aberration and bacterial reverse mutation assays) and acute oral toxicity assays were conducted. In the in vitro genotoxicity assays, bacterial reverse mutation and chromosomal aberration assays showed that BLE0 possessed no mutagenicity or clastogenicity. Furthermore, the median lethal dose (LD50) of BLE0 was higher than 5000 mg/kg in female and male Sprague-Dawley (SD) rats and no adverse effects were observed in terms of mortality and abnormal changes in clinical signs (body weight and necropsy). Based on these results, BLE0 was found to be safe with regards to genotoxicity under our test conditions, demonstrating no acute oral toxicity up to 5000 mg/kg in SD rats.

Phosphorylation of PTEN plays an important role in carcinogenesis and progression of gastric cancer. However, the underlying mechanism of PTEN phosphorylation regulation remains largely elusive. In the present study, PDZK1 was identified as a novel binding protein of PTEN by association of PTEN through its carboxyl terminus and PDZ domains of PDZK1. By direct interaction with PTEN, PDZK1 inhibited the phosphorylation of PTEN at S380/T382/T383 cluster and further enhanced the capacity of PTEN to suppress PI3K/AKT activation. PDZK1 suppressed gastric cancer cell proliferation by diminishing PI3K/AKT activation via inhibition of PTEN phosphorylation in vitro and in vivo. The expression of PDZK1 was frequently downregulated in gastric cancer specimens and correlated with progression and poor prognosis of gastric cancer patients. Downregulation of PDZK1 was associated with PTEN inactivation, AKT signaling and cell proliferation activation in clinical specimens. Thus, low levels of PDZK1 in gastric cancer specimens lead to increase proliferation of gastric cancer cells via phosphorylation of PTEN at the S380/T382/T383 cluster and constitutively activation of PI3K/AKT signaling, which results in poor prognosis of gastric cancer patients.

The edible and medicinal perennial herb Aster scaber is known to have anticancer, antioxidant, and immunomodulatory properties. However, the biological effects of its polysaccharides are not well understood. Here, we aimed to extract novel polysaccharides with enhanced biological properties from Aster scaber using enzyme-assisted methods. Amylase, cellulase, and pectinase were used to extract enzyme-assisted polysaccharide (ASEP)-A, ASEP-C, and ASEP-P, respectively. The yields, physicochemical properties, and immunostimulatory activities of the polysaccharides were investigated and compared with those of hot water extracted polysaccharide (ASWP). The highest yield (3.8%) was achieved for ASEP-P extracted using pectinase digestion. Fourier-transform infrared spectroscopy (FT-IR) and chemical composition analysis revealed that ASWP and three ASEPs were typical acidic heteropolysaccharides, mainly comprising rhamnose, arabinose, galactose, glucose, and galacturonic acid. Immunostimulatory activity assays on RAW264.7 macrophages showed ASEP-P to have the greatest immunostimulatory potential in terms of nitric oxide (NO) and cytokine productions and phagocytic activity. ASEP-P administration improved immune-enhancing effects in normal mice by improving the spleen index and splenic lymphocyte proliferation, and in immunosuppressed mice by modulating lymphocyte proliferation, natural killer (NK) cell activity, and leukocyte counts. The ASEP-P derived from pectinase hydrolysate of Aster scaber demonstrated efficacious immunostimulatory properties and has potential applications as an immune stimulator.

Innate cytokines are critical drivers of priming and differentiation of naive CD4 T cells, but their functions in memory T cell response are largely undefined. Here we show that IL-1 acts as a licensing signal to permit effector cytokine production by pre-committed Th1 (IFN-γ), Th2 (IL-13, IL-4, and IL-5) and Th17 (IL-17A, IL-17F, and IL-22) lineage cells. This licensing function of IL-1 is conserved across effector CD4 T cells generated by diverse immunological insults. IL-1R signaling stabilizes cytokine transcripts to enable productive and rapid effector functions. We also demonstrate that successful lineage commitment does not translate into productive effector functions in the absence of IL-1R signaling. Acute abrogation of IL-1R signaling in vivo results in reduced IL-17A production by intestinal Th17 cells. These results extend the role of innate cytokines beyond CD4 T cell priming and establish IL-1 as a licensing signal for memory CD4 T cell function., CD4 T cell polarizations and functions are regulated by cytokines from innate cells. Here the authors show that IL-1 deficiency does not impair the differentiation of Th1, Th2 and Th17, but IL-1 signaling is required for maintaining the expressions of their respective key cytokines to ‘license’ the functions of these T cell subsets.

Background Systemic lupus erythematosus (SLE) is characterized by the presence of multiple autoantibodies as well as clinical evidence of multi-system immune-mediated pathology. Individuals who develop SLE typically demonstrate serological autoimmunity for several years prior to the onset of clinical disease. Measuring anti-nuclear antibodies (ANA) is a common test for humoral autoimmunity. It is sensitive (95+%) for SLE, but not specific as up to 25% of healthy controls (HC) will have a measurable ANA. While very few of the ANA +HC will develop SLE, they represent a group at higher risk of the disease. Understanding the risks for ANA positivity provides essential knowledge about the development of SLE. Methods Serum and DNA were collected from 2903 healthy individuals with no personal history of autoimmunity. Antinuclear antibodies were detected using Inova QuantaLite ELISA. Sera from subset (n=724) individuals (ANA− HC, ANA+ HC, and SLE) were assayed by protein microarray quantifying IgM and IgG responses to 89 previously known human autoantigens. A nested cohort consisting of all the ANA +Caucasian individuals and age/gender matched ANA− controls were genotyped using the ImmunoChipv.1 SNP array. Results In HC, 16% had moderate and 10% had high levels of IgG ANA. Autoantigen microarray data showed that ANA+ HC had a high prevalence of antibodies to non-nuclear and cytoplasmic antigens while subjects with SLE predictably produced antibodies to a variety of nuclear antigens as well. A quantitative genetic association test with ANA identified the locus c11orf30 or EMSY, associated with high ANA phenotype in the healthy population (see figure 1). This locus codes for a negative transcriptional regulator. A haplotype comprised of many potentially regulatory polymorphisms at EMSY contributed to strong risk for ANA in healthy individuals [(p=3.83E-04, OR=2.6)]. eQTL data suggests that the ANA-associated EMSY haplotype leads to reduced expression of EMSY protein in human macrophages and EBV cell lines. Autoantibody profiles of serum samples from healthy individuals with the EMSY risk genotype exhibited high titers of anti-IgG and IgA antibodies targeted to multiple autoantigens as well as food and environmental allergens. Conclusions EMSY, a locus previously linked with atopy, psoriasis and inflammatory bowel disease was associated with ANA in healthy individuals. The EMSY protein is a BRCA2-associated transcriptional repressor. Individuals with risk haplotypes in EMSY make a wide variety of disease-associated antibodies, suggesting an early common pathway for autoimmune and allergic conditions.

Background The antibody specificities of an infant progressively form in response to infections, environmental exposures, and vaccinations. While many adults develop antibodies to self-antigens, it remains unknown if these are present in infants and toddlers. Methods Serum, peripheral blood, and clinical data have been collected from 102 healthy children (1–2 year of age). ANA titers were measured by QuantiaLite ELISA (Inova) and reactivity to 125 diverse autoantigens tested by autoantibody array. Targeted sequencing was performed in 60 children to capture HLA alleles and potentially pathogenic genetic variations in 100 plus loci that are implicated in various autoimmune, rheumatic and immune system related diseases. Sequencing libraries were made using KAPA Biosystems kits. Custom target oligos were synthesized from Nimblegen. Deep sequencing was performed using Illumina HiSeq 4000 platform. Sequencing reads were aligned to reference genome and variants were called using GATK Pipeline. Secondary data analysis is done using Metlab, GraphPad Prism, Haploview and Golden Helix programs. Results Approximately 28% of very young children have moderate to high-titer autoantibodies, similar to adults, however, no female gender bias was observed. Significant differences between the child and adult immune repertoires were seen. Some samples demonstrated strong signatures of non-nuclear antigens reactivities. Interestingly, the ANA positive group of children exhibited significantly high titers of anti-alpha fodrin IgG (p=0.01), an autoantibody reported in juvenile Sjogren’s syndrome. Analysis of sequencing data identified regulatory polymorphisms in HLA class II and III regions that were associated with ANA positivity. About 8% of the ANA positive children also carried autoimmune disease associated HLA-DR-DQ alleles. HLA alleles and regulatory haplotypes were analyzed in relation to various clinical features in children. Serum C4 level measured in subset of ANA positive children identified few with reduced expression. Conclusions While the immune repertoire of very young children is typically thought of as naive and self-tolerant, a significant fraction of very young children makes autoantibodies as detected by commercial ANA ELISA and autoantigen arrays. This supports the conclusion that ANA and other autoantibodies are consequences of general body development and immune upregulation and not markers of pathology. One of the major genetic determinants of ‘pre-clinical’ autoimmunity as measured by ANA is the human major histocompatibility locus, HLA. Within HLA, several ‘endophenotypes’ emerge, including expression of a complement protein involved in immune complex removal, as well as multiple proteins associated with antigen presentation, including MHC class II. These findings support the idea that there is quantifiable genetic risk for the development of autoimmunity that can be measured in very young individuals.

In this study, a crude water-soluble polysaccharide (LLWP-C) was extracted from lotus leaves and further purified by size exclusion chromatography, to obtain the two main polysaccharides, LLWP-1 and LLWP-3. Physical and chemical analyses showed that they were homogeneous polysaccharides in β-type glycosidic linkage. LLWP-1 was devoid of helical conformation, had a molecular weight of 85.1 kDa and was mainly composed of Rha, Ara, Gal, Glu, and GalA in a molar ratio of 7.0:24.8:28.0:6.0:26.4. LLWP-3 showed a helical conformation, had a molecular weight of 12.5 kDa and consisted mainly of Rha, Ara, Gal, Glu, Man, and GalA in a molar ratio of 6.6:9.8:15.0:8.9:6.1:47.2. It was demonstrated that LLWP-C and both purified LLWP-1 and LLWP-3 could effectively enhance the proliferation, phagocytosis, nitric oxide (NO), and cytokine secretions by activating corresponding mRNA expression in macrophages, via MAPK and NF-κB pathways. LLWP-3 displayed the greatest immunostimulatory potential, followed by LLWP-1 and LLWP-C. These findings suggest that polysaccharides extracted from lotus leaf exert immunostimulatory activity that could be further investigated to develop functional foods and natural immunopotentiating therapeutic agents.

Persimmon (Diospyros kaki L.f.) leaves have traditionally been used as a phytomedicine, in health beverages to treat cardiovascular and respiratory disease and to promote maternal health in East Asia. In particular, polysaccharides from persimmon are known to have anti-coagulant, anti-oxidant, and immune-stimulatory activities. However, their beneficial effects against osteoporosis have not been reported. In the present study, we investigated the anti-osteoporotic effects of polysaccharides from persimmon leaves (PLE0) using an in vivo model of ovariectomy (OVX)-induced bone loss and an in vitro system of receptor activator of nuclear factor-&kappa, B ligand (RANKL)-induced osteoclast differentiation. In the OVX mouse model, PLE0 remarkably improved OVX-induced trabecular bone loss by suppressing osteoclast activity. In primary bone marrow-derived macrophages (BMMs), PLE0 dose-dependently inhibited osteoclast differentiation. In addition, PLE0 down-regulated RANKL-induced activation of mitogen-activated protein kinases (MAPKs) such as p38, ERK, and JNK resulting in suppression of nuclear factor of activated T cells cytoplasmic 1 (NFATc1) expression. Our results indicate that PLE0 has anti-osteoporotic effects in OVX-induced bone loss via inhibition of osteoclast differentiation. Taken together, PLE0 from persimmon may prevent postmenopausal bone loss and osteoporotic bone fragility.

Cervical cancer is one of the most lethal types of cancer in female. Aberrant activation of Wnt/β-catenin signaling pathway has been found to be involved in cervical cancer development and progression, whereas the underlying molecular mechanisms remain poorly understood. The present study showed that NHERF1 was a novel gene associated with both cell proliferation and Wnt signaling pathway in cervical cancer by analysis of differential gene expression and gene cluster for the cervical cancer specimens from GEO data sets. It was further demonstrated in cellular study that NHERF1 inhibition of cervical cancer cell proliferation through Wnt/β-catenin signaling was dependent on α-actinin-4 (ACTN4) expression. A negative association between NHERF1 expression and levels of ACTN4 and β-catenin was found in mouse xenograft model and cervical cancer specimens. Low levels of NHERF1 in cervical cancer specimens were found to associate with activation of cell proliferation and Wnt/β-catenin signaling by gene set enrichment analysis, and also were an independent predictive factor for worse prognosis of cervical cancer patients by Cox regression analysis. These findings demonstrate that NHERF1 inhibits Wnt signaling-mediated proliferation of cervical cancer via suppression of ACTN4, and NHERF1 downregulation may contribute to the progression of cervical cancer. These findings may also shed some lights for understanding the underlying mechanisms of cisplatin resistance and worse prognosis of HPV-inactive cervical cancer patients.

Gliomas are the most common primary brain malignancies and are associated with a poor prognosis. Here, we showed that the PDZ domain-containing protein membrane-associated guanylate kinase inverted 3 (MAGI3) was downregulated at the both mRNA and protein levels in human glioma samples. MAGI3 inhibited proliferation, migration, and cell cycle progression of glioma cells in its overexpression and knockdown studies. By using GST pull-down and co-immunoprecipitation assays, we found that MAGI3 bound to β-catenin through its PDZ domains and the PDZ-binding motif of β-catenin. MAGI3 overexpression inhibited β-catenin transcriptional activity via its interaction with β-catenin. Consistently, MAGI3 overexpression in glioma cells C6 suppressed expression of β-catenin target genes including Cyclin D1 and Axin2, whereas MAGI3 knockdown in glioma cells U373 and LN229 enhanced their expression. MAGI3 overexpression decreased growth of C6 subcutaneous tumors in mice, and inhibited expression of β-catenin target genes in xenograft tumors. Furthermore, analysis based on the Gene Expression Omnibus (GEO) glioma dataset showed association of MAGI3 expression with overall survival and tumor grade. Finally, we demonstrated negative correlation between MAGI3 expression and activity of Wnt/β-catenin signaling through GSEA of three public glioma datasets and immunohistochemical staining of clinical glioma samples. Taken together, these results identify MAGI3 as a novel tumor suppressor and provide insight into the pathogenesis of glioma.

Phosphatase of regenerating liver-3 (PRL-3) has been reported to have a critical role in metastatic progression of cancers. Here, we investigate how PRL-3 increases the malignant degree of melanoma cells. The expression of PRL-3 increased gradually during the malignant progression of melanoma. The phosphorylation of Akt was elevated in highly malignant melanoma cells, which was accompanied by a decrease in nuclear phosphatase and tensin homolog (PTEN). The phosphorylation of NHERF1 in the serine site was regulated by PRL-3 and showed cytoplasmic translocation upon dephosphorylation, which resulted in a decrease in nuclear PTEN. The co-translocation of NHERF1 and PTEN from the nucleus to the cytoplasm was observed during the malignant progression of melanoma cells. Tumor growth was inhibited significantly, and the survival was prolonged upon knockdown of cytoplasmic NHERF1 in B16BL6 cells prior to the inoculation into mice. Taken together, to our knowledge previously unreported, we have identified NHERF1 as a potential substrate of PRL-3. Its phosphorylation status as well as its change in cellular localization and association with PTEN correlated with the malignant progression of melanoma. Our data provide an explanation for how PRL-3 promotes the malignant progression of melanoma, as well as a diagnostic marker or therapeutic target for malignant melanoma.

Flavor development in soy sauce is significantly related to the diversity of yeast species. Due to its unique fermentation with meju, the process of making Korean soy sauce gives rise to a specific yeast community and, therefore, flavor profile; however, no detailed analysis of the identifying these structure has been performed. Changes in yeast community structure during Korean soy sauce fermentation were examined using both culture-dependent and culture-independent methods with simultaneous analysis of the changes in volatile compounds by GC-MS analysis. During fermentation, Candida, Pichia, and Rhodotorula sp. were the dominant species, whereas Debaryomyces, Torulaspora, and Zygosaccharomyces sp. were detected only at the early stage. In addition, Cryptococcus, Microbotryum, Tetrapisispora, and Wickerhamomyces were detected as minor strains. Among the 62 compounds identified in this study, alcohols, ketones, and pyrazines were present as the major groups during the initial stages, whereas the abundance of acids with aldehydes increased as the fermentation progressed. Finally, the impacts of 10 different yeast strains found to participate in fermentation on the formation of volatile compounds were evaluated under soy-based conditions. It was revealed that specific species produced different profiles of volatile compounds, some of which were significant flavor contributors, especially volatile alcohols, aldehydes, esters, and ketones.

Changes in immune function during the course of systemic lupus erythematosus (SLE) are well characterized. Class-switched antinuclear antibodies are the hallmark of SLE, and T/B-cell interactions are thus critical. However, changes in immune function contributing to disease susceptibility are unknown. Here, we have analyzed primary T and B cells from a mouse model of SLE prior to the onset of disease. To allow cognate T-cell activation with low affinity, we have developed a lower potency peptide ligand for the OTII TCR. T- and B-cell couples formed less frequently and retained their polarity less efficiently preferentially in response to low-affinity stimulation in SLE-prone mice. This matched decreased recruitment of actin and Vav1 and an enhanced PKCΘ recruitment to the cellular interface in T cells. The induction of the GC B-cell marker GL7 was increased in T/B cell couples from SLE-prone mice when the T-cell numbers were limited. However, the overall gene expression changes were marginal. Taken together, the enhanced cell-couple transience may allow a more efficient sampling of a large number of T/B cell couples, preferentially in response to limiting stimuli, therefore enhancing the immune reactivity in the development of SLE.

Two lactic acid bacteria (LAB), Lactobacillus homohiochii JBCC 25 and L. homohiochii JBCC 46, were isolated from a naturally fermented vinegar beverage. Fermented pepper leaves beverage (FPLB-3) which indicated good assessment based on a sensory evaluation test was prepared using the mixed strains. The prepared FPLB-3 showed enhanced functional aspects based on γ-aminobutyric acid (23.6 mg%), total polyphenol (2,828.2 gallic acid equivalents μg/g), and flavonoid contents (2,008.8 catechin equivalents μg/g). Moreover, FPLB-3 showed significantly stronger scavenging activities for the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (84.6%) and superoxide radical (90.8%) and outstanding inhibitory activity against α-glucosidase of 65.9% at 50 mg/mL compared to the raw material. Based on the results, the newly developed FPLB using Wongi-1 pepper leaves and Lactobacillus is expected to be a novel functional food with natural antioxidant and antidiabetic capacities.

The vent sexing and the auto-sexing by using sex-linked traits are general sexing methods of day-old chicks. Currently, the feather sexing which is based on the differences in the feather characteristics at hatching is the representative sexing method of chicken, because the late-feathering is sex-linked trait. The feather sexing can be used if the breed has dominant feathering gene (K) in maternal and recessive gene (k + ) in paternal. Therefore it is necessary to identify the associa- tion of feathering genes and quantitative traits in chickens. In this study, we investigated the influence of the rate of feathering on productive traits in Korean Native Chicken. In results, there was no significant difference between early-feathering chickens and late-feathering chickens in reproductive performance such as fertility and hatchability. Livability, body weights, egg production, egg weight and egg quality also did not significantly differ between early- and late-feathering chickens. Age at first egg was the only trait of those tested in which significant difference was observed. The early-feathering chickens laid eggs 3 days earlier than late-feathering chicken. As a result, there is no influence of feathering phenotypes on productive performance in Korean Native Chickens. Consequentially, establishing the feather sexing strain is available using the Korean Native Chicken breed without considering of the effect of feathering genes on productive traits.

Osteopontin (OPN) is known to be significantly involved in the pathogenesis of rheumatoid arthritis (RA). This study aimed to evaluate if the serum concentration of OPN in patients with RA before and after therapeutic treatments was correlated to disease activity and response to therapy. Blood samples from 40 patients with RA were collected at baseline and six months after starting treatment with disease-modifying antirheumatic drugs (DMARDs) and/or tumor necrosis factor (TNF)-α blockers. Serum levels of OPN were measured by ELISA. At baseline, the serum OPN level in RA patients was significantly higher than that of the healthy group. The OPN level at baseline in RA patients with severe disease activity as evaluated by DAS28 was slightly higher than that of those with moderate disease activity. The serum OPN level in RA patients was not significantly correlated with the DAS28 level. The serum OPN level in both responders and non-responders after therapy was significantly decreased regardless of responsiveness to therapy. Also, the OPN level at baseline did not affect the responsiveness to therapeutic treatments. In conclusion, serum OPN level was not correlated with disease activity or responsiveness of RA patients to therapeutic treatments.

This work is aimed to increase knowledge of the functional exopolysaccharide (EPS) from lactic acid bacteria (LAB) in makgeolli, a Korean fermented rice wine. Among LAB strains isolated from makgeolli, strain M76 was selected as a functional strain producing a bioactive EPS, based on its antioxidative activity on the DPPH radical. The 16S rRNA gene sequencing analysis showed a high sequence similarity (99.0%) with P. acidilactici, but had different biochemical properties with the already known P. acidilactici type strains in the aspect of carbohydrates utilization. The obtained P. acidilactici M76 produced a soluble EPS above 2 g/l. One-step chromatography using gel filtration after ethanol precipitation from the supernatant of P. acidilactici M76 was enough to obtain purified EPS with a single peak, showing a molecular mass of approximately 67 kDa. Componential and structural analyses of EPS by TLC, HPLC, and FT-IR indicated that the EPS is a glucan, consisting of glucose units. The purified EPS had antioxidant activity on the DPPH radical of 45.8% at a concentration of 1 mg/ml. The purified EPS also showed proliferative effect on the pancreatic RIN-m5F cell line and remarkable protection activity on alloxan-induced cytotoxicity. This potent antioxidant and antidiabetic EPS by LAB in makgeolli may contribute to understanding the functionality of makgeolli.

A strain producing exopolysaccharide (EPS) with strong hydroxyradical scavenging activity and antityrosinase activity has been isolated previously. However, the EPS production rate of the strain (7.3 g/L) was relatively low to utilize it at the industrial level. Therefore, in this study, optimal medium and fermentation conditions were examined to enhance EPS production by Bacillus licheniformis KS-17. Maximum EPS production was obtained in medium with 125 g/L sucrose, 30 g/L ammonium sulfate, and 10 mM calcium chloride without a phosphate source at 37°C for 5 days with the initial pH adjusted to 8.5. Under optimal culture conditions, EPS was produced at up to 27.2 (at 5 days) vs. 12.0 g/L (at 7 days under basal conditions), which was 2.3 times greater and in a shorter time than the production yield possible without optimizing conditions.

G protein-coupled estrogen receptor (GPER) signaling is activated in triple-negative breast cancer (TNBC); however, the detailed mechanisms of its regulation remain unclear. The present study aimed to elucidate the molecular mechanisms involved in GPER activation in TNBC. In MDA-MB-231 cells, a TNBC cell line, NHERF1 interaction with GPER was verified by co-immunoprecipitation and immunofluorescent staining assays. Overexpression of NHERF1 in MDA-MB-231 cells inhibited GPER-mediated proliferation and phosphorylation of ERK1/2 and Akt. Furthermore, NHERF1 expression levels were negatively correlated with the gene signatures of GPER activation, ERK1/2 and Akt signaling, and cell proliferation in early stage of TNBC tumors from the TCGA data set. Taken together, NHERF1 inhibited the activation of GPER-mediated signaling and suppressed the proliferation of triple-negative breast cancer cells. Loss of NHERF1 expression may play a pivotal role in the early stage of TNBC carcinogenesis.

The human immune system defends the body against microbes and other threats. However, if this process goes wrong the immune system can attack the body’s own healthy cells, which can lead to serious autoimmune diseases. Systemic lupus erythematosus (SLE) is an autoimmune disease in which immune cells often attack internal organs – including the kidneys, nervous system and heart. Over the past decade, multiple genes have been linked with an increased risk of SLE. However, it is largely unknown how the sequences of these genes differ between individuals with SLE and healthy individuals, and the precise changes that lead to an increased risk of SLE are also not clear. Now, Raj, Rai et al. have determined the genetic sequences of over 700 people with SLE and over 500 healthy individuals and looked for differences that influence susceptibility to the disease. The vast majority of differences were discovered in stretches of DNA that regulate the expression of nearby genes, rather than in DNA that encodes the structures of proteins. Notably, extensive differences were found in a region of the human genome that regulates the production of proteins called Human Leukocyte Antigen class II molecules; which are known to play a critical role in activating the immune system. Raj, Rai et al. found that slight changes to the regulatory DNA sequences resulted in an overabundance of these proteins, which led to a hyperactive immune system that is strongly associated with SLE. Future studies could now ask if the changes to the regulatory DNA sequences highlighted by Raj, Rai et al. increase susceptibility to other autoimmune disorders as well. It may also be possible to use the increased understanding of how the immune system is regulated to develop new ways to minimize the rejection of organ transplants.

Objectives To identify non-major histocompatibility complex susceptible genes that might contribute to Behcet9s disease (BD). Methods We performed a genome-wide association study using DNA samples from a Korean population consisting of 379 BD patients and 800 controls. A replication study was performed in a Japanese population (363 BD patients and 272 controls). To evaluate the functional implication of the target single nucleotide polymorphisms (SNP), gene expression levels in peripheral T cells, allele-specific modulation of promoter activity and biological effect of mRNA knockdown were investigated. Results We found a novel association of BD to the GIMAP locus, mapped to chromosome 7q36.1 (rs1608157, p=6.01×10 −8 in a minor allele dominant model; rs11769828, allele based p=1.60×10 −6 ). A fine mapping study identified an association with four additional SNP: rs1522596 (OR=1.45, p=7.70×10 −6 ) in GIMAP4 ; rs10266069 (OR=1.32, p=2.67×10 −4 ) and rs10256482 (OR=1.27, p=5.27×10 −4 ) in GIMAP2 ; and rs2286900 (OR=1.61, p=3.53×10 −5 ) in GIMAP1 areas. Replication study using DNA samples from the Japanese population validated the significant association between BD and the GIMAP locus. The GIMAP4 promoter construct plasmid with the minor allele of rs1608157 displayed significantly lower activity than one with the major allele. Moreover, CD4 T cells from BD patients showed a lower level of GIMAP4 mRNA, and GIMAP4 knockdown was protective against Fas-mediated apoptosis. Conclusions These results suggest that a GIMAP cluster is a novel susceptibility locus for BD, which is involved in T-cell survival, and T-cell aberration can contribute to the development of BD.

Oxygen generating (OG) hydrogels immobilizing glucose oxidase (GOx, EC 1.1.3.4) and peroxidase (PO) (EC 1.11.1.7) were prepared using a γ-ray irradiation polymerization technique, and the properties were examined. In physiological tests of the OG hydrogels ranging from 15 to 70 kGy, GOx or PO showed higher gel contents at 87% for the poly(vinyl alcohol) (PVA) hydrogel and 90% for the poly(vinylpyrrolidone) (PVP) hydrogel. The optimal irradiation conditions affecting the entrapment ratio for preparing the OG hydrogel was 35 kGy for the PVP hydrogel, and both enzymes (GOx, 95% and PO, 89%). Optimal GOx and PO activity of the prepared OG hydrogel occurred at pH 6.0 and 35 °C, and both enzymes showed stable activity under acidic conditions. The optimal glucose condition was 4 mg, which resulted in a maximum oxygen generating yield by the OG hydrogels. When oxygen concentration was compared, the OG hydrogels showed higher oxygen pressure than that of control hydrogels without the GOx and PO enzymes. Moreover, the OG hydrogel continuously converted potassium iodide to iodine in a glucose dosage and time dependant manner for 15 h, producing approximately 0.1 M iodine in 5 h.

The method of sexing based on differences in the rate of feather growth provides a convenient and inexpensive approach. The locus of feather development gene (K) is located on the Z chromosome and can be utilized to produce phenotypes that distinguish between the sexes of chicks at hatching. To establish the auto-sexing native chicken strains, this study analyzed the genotype frequency of the feathering in domestic chicken breeds. The method of classification of slow- and rapid-feathering chickens was also investigated. In the slow-feathering chicks, the coverts were either the same length or longer than the primary wing feathers at hatching. However, the rapid-feathering chicks had the primary wing feathers that were longer than the coverts. The growth pattern of tail feather also distinctively differed between the rapid- and slow-feathering chicks after 5-days. The accuracy of wing feather sexing was about 98% compared with tail sexing. In domestic chicken breeds, Korean Black Cornish, Korean Rhode Island Red, and Korean Native Chicken-Red had both dominant (K) and recessive () feathering genes. The other breeds of chickens, Korean Brown Cornish, Ogol, White Leghorn, Korean Native Chicken-Yellow, -Gray, -White and -Black had only the recessive feathering gene (). Consequently, feather sexing is available using the domestic chicken breeds. Establishing the maternal stock with dominant gene (K-) and paternal stock with recessive gene (), the slow-feathering characteristic is passed from mothers to their sons, and the rapid-feathering characteristic is inherited by daughters from their fathers.

Improvement of natamycin production by Streptomyces gilvosporeus ATCC 13326 was performed by recursive protoplast fusion in a genome-shuffling format. After four rounds of genome shuffling, the best producer, GS 4-21, with genetic stability was obtained and its production of natamycin reached 4.69 ± 0.05 g/L in shaking flask after 96 h cultivation, which was increased by 97.1% and 379% in comparison with the highest parental strain pop-72A(r)07 and the initial strain ATCC 13326, respectively. Compared with the initial strain ATCC 13326, the recombinant GS 4-21 presented higher polymorphism. Fifty-four proteins showed differential expression levels between the recombinant GS 4-21 and initial strain ATCC 13326. Of these proteins, 34 proteins were upregulated and 20 proteins were downregulated. Of the upregulated proteins, one protein, glucokinase regulatory protein, was involved in natamycin biosynthesis. This comprehensive analysis would provide useful information for understanding the natamycin metabolic pathway in S. gilvosporeus.

This study examined the efficient production and optimal separation procedures for pure L-threo-3,4-dihydroxyphenylserine (L-threo-DOPS) from a mixture of diastereomers synthesized by whole-cell aldol condensation reaction, harboring diastereoselectivity-enhanced L-threonine aldolase in Escherichia coli JM109. The addition of the reducing agent sodium sulfite was found to stimulate the production of L-threo-DOPS without affecting the diastereoselectivity ratio, especially at the 50 mM concentration. The optimal pH for diastereoselective synthesis was 6.5. The addition of Triton X-100 also strongly affected the synthesis yield, showing the highest conversion yield at a 0.75% concentration; however, the diastereoselectivity of the L-threonine aldolase was not affected. Lowering the temperature to 10°C did not significantly affect the diastereoselectiviy without affecting the synthesis rate. At the optimized conditions, a mixture of L-threo-DOPS and L-erythro-DOPS was synthesized by diastereoselectivity-enhanced L-threonine aldolase from E. coli in a continuous process for 100 hr, yielding an average of 4.0 mg/mL of L-threo-DOPS and 60% diastereoselectivity (de), and was subjected to two steps of ion exchange chromatography. The optimum separation conditions for the resin and solvent were evaluated in which it was found that a two-step process with the ion-exchange resin Dowex 50 W × 8 and activated carbon by washing with 0.5 N acetic acid was sufficient to separate the L-threo-DOPS. By using two-step ion-exchange chromatography, synthesized high-purity L-threo-DOPS of up to 100% was purified with a yield of 71%. The remaining substrates, glycine and 3,4-dihydroxybenzaldehyde, were recovered successfully with a yield of 71.2%. Our results indicate this potential procedure as an economical purification process for the synthesis and purification of important L-threo-DOPS at the pharmaceutical level.

This experiment was carried out in order to collect the basic data on the standardization of the manufactur- ing process of Rehmannia glutinosa Libosch. var. purpurea Makino drying. By the drying methods of R. glutinosa, the con- tent of water, inorganic components, reducing sugar, catalpol and benzo(α)pyrene were investigated. The water content was 15.6~17.2% when R. glutinosa was dried by cold-warm air moisture absorption drying method (CAMAD) at 60℃ during 6 days. Among of the inorganic components of R. glutinosa the K content was the most followed by P, Na, Ca and Mg. The reducing sugar content of R. glutinosa by the hot air drying method (HAD) was much more than that by the CAMAD. The catalpol content of R. glutinosa was not different by the drying temperature when it was dried by the CAMAD. The catalpol content of the large size tuber (about 50.0 g/unit) showed a tendency to increase from 60℃ until 70℃ drying temperature, but that of the small size tuber(about 4.0 g/unit) was decreased as being a trend as the drying temperature high when R. glu- tinosa was dried by the HAD, But the catalpol content R. glutinosa had a tendency to drop significantly at drying tempera- ture above 80℃. The benzo(α)pyrene content was little detected when R. glutinosa was dried by both the SLD and the CAMAD, and the sampling by the HAD indicated within the scope of 5 ㎍/㎏ which was the scope to regulate by Korean food and drug administration. In conclusion, it seemed that an appropriate drying temperature of R. glutinosa by the CAMAD and the HAD was about 60℃ and about 70℃, respectively, when we consider the catalpol content and benzo(α)pyrene detection in the manufacturing process of drying R. glutinosa.

Fermented sugar-soaked black soybean snacks (FSBSS) were prepared using optimized conditions at 2 times sugar soaking process to give a desirable form and texture after fermentation with Lactobacillus plantarum (LP) and Bacillus subtilis (BS) for 48 h. The prepared FSBSS showed higher total polyphenol and flavonoid contents than an unfermented (NF) sample; total isoflavone content increased approximately 31 and 17% (LP and BS, respectively), as compared to NF. γ-Aminobutyric acid was also contributed by LP and BS, indicating significantly higher contents of 16.9 and 21.6 mg/g (LP and BS, respectively) compared to NF (15.2 mg/g). Moreover, the FSBSS showed antioxidant activities on DPPH, superoxide, and hydroxyl radicals as compared to NF. Based on the results, the newly developed FSBSS using solid-fermentation indicate a useful and convenient way to replace traditional soybean products from the standpoint that also provides an easy means to supply high nutritive and functional properties.

The eutrophication of water in an urban river was treated by combining two submerged aquatic plantsElodeanuttalliiandPotamogeton crispuswith a biological growth-promoting agent Bio-energizer (BE) on a pilot scale. The results showed that the two submerged plants could remove CODCr effectively and steadily, with a removal rate of 58% and 52.3% respectively. All experimental treatments were effective at removing TN, TP, NH4+-N, NO3+-N, but the treatment in which BE was added at the lower concentration (from 1 to 3 ml/m3) was better than that at the higher concentration (from 5 to 10 ml/m3).Elodea nuttalliiproduced the best treatment efficiency when 1 ml/m3BE was added to the experimental water. The rate of removal of TP and TN was 71.0%, and 72.4% within one week while that in the control group was 50% and 51% respectively. The number of bacteria changed during the different stages of the experiment.

Human liposarcoma is the most common soft tissue sarcoma. There is no effective therapy so far except for surgery. In this study, we report for the first time that curcumin induces endoplasmic reticulum (ER) stress in human liposarcoma cells via interacting with sarcoplasmic/endoplasmic reticulum Ca2+-ATPase 2 (SERCA2). Curcumin dose-dependently inhibited the cell survival of human liposarcoma cell line SW872 cells, but did not affect that of human normal adipose-derived cells. Curcumin-mediated ER stress via inhibiting the activity of SERCA2 caused increasing expressions of CHOP and its transcription target death receptor 5 (TRAIL-R2), leading to a caspase-3 and caspase-8 cascade-dependent apoptosis in SW872 cells in vitro and in vivo. Moreover, 70% of human liposarcoma tissues showed an elevated SERCA2 expression compared with normal adipose tissues. Curcumin dose-dependently inhibited the activity of SERCA2, and the interaction of molecular docking and colocalization in ER of curcumin with SERCA2 were further observed. These findings suggest that curcumin may serve as a potent agent for curing human liposarcoma via targeting SERCA2. Mol Cancer Ther; 10(3); 461–71. ©2011 AACR.

Several approaches have been used to encourage the differentiation of cardiomyocytes from human embryonic stem cells. However, the differentiation efficiency is low, and appropriate culture protocols are needed to produce adequate numbers of cardiomyocytes for therapeutic cell transplantation. This study investigated the effects of serum on cardiomyocyte differentiation in suspension culture medium during embryoid body (EB) formation by human embryonic stem cells. The addition of ascorbic acid, dimethylsulfoxide and 5-aza-2'-deoxycytidine during days 5-7 at the EB-forming stage resulted in an increase in the numbers of rhythmically contracting clusters of derived cardiomyocytes. Treatment with 0.1 mmol L(-1) ascorbic acid alone, or more notably in combination with 10 micromol L(-1) 5-aza-2'-deoxycytidine, induced the formation of beating cells within EBs. Most of the beating clusters had spontaneous contraction rates similar to those found in human adults, and their contractile activity lasted for up to 194 days.