Characterization of a Small Cysteine-Rich Secreted Effector, TcSCP_9014, in Tilletia controversa.

Tilletia controversa J. G. Kühn is the causal agent of wheat dwarf bunt (DB), a destructive disease causing tremendous economic losses. Small cysteine-rich secreted proteins (SCPs) of plant fungi are crucial in modulating host immunity and promoting infection. Little is known about the virulence effectors of T. controversa. Here, we characterized TcSCP_9014, a novel effector of SCPs, in T. controversa which suppressed programmed cell death triggered by BAX without relying on its signal peptide (SP). The SP in the N-terminus of TcSCP_9014 was functional in the secretory process. Live-cell imaging in the epidermal cells of Nicothiana benthamiana suggested that TcSCP_9014 localized to the plasma membrane, cytoplasm, and nucleus. Furthermore, yeast cDNA library screening was performed to obtain the interacting proteins in wheat. Yeast two-hybrid and BiFC assays were applied to validate the interaction of TcSCP_9014 with TaMTAN and TaGAPDH. Our work revealed that the novel effector TcSCP_9014 is vital in modulating plant immunity, which opens up new avenues for plant-pathogen interactions in the T. controversa infection process.

Assessment and management of radiation-induced trismus in patients with nasopharyngeal carcinoma: a best practice implementation project.

INTRODUCTION AND AIMS: Intensity-modulated radiotherapy (IMRT) is the most commonly used radiotherapy technology in oncology, which enables precise conformation of the radiation dose to the target volume and reduces the risk of radiation damage to the adjacent normal structures. Nevertheless, it is still inevitable for IMRT of head and neck cancer to cause radiation-related toxic and side effects, such as dry mouth, mucositis, oral dysarthria, taste disorder, osteonecrosis, and trismus. Trismus is one of the most common late side effects caused by radiotherapy of nasopharyngeal carcinoma (NPC), which seriously affects the quality of life for patients with NPC. However, the current clinical assessment and management of trismus after radiotherapy for NPC are still imperfect. This best practice implementation project aimed to implement an evidence-based practice in assessing and managing trismus for NPC patients who underwent radiotherapy, thereby improving the compliance of clinical practice with the best evidence and the quality of life of patients with NPC. METHODS: This evidence-based audit and feedback project was implemented using a three-phase approach at a third-class hospital in China, following JBI's Practical Application of Clinical Evidence System (PACES) and GRiP evidence application. The first phase included a baseline audit with six evidence-based audit criteria derived from the best available evidence. The second phase included analyzing the results of the baseline audit, identifying barriers to compliance with best practice principles, and developing and implementing strategies to address the barriers identified in the baseline audit. The third phase involved a follow-up audit to assess the results of the interventions implemented to improve practice. RESULTS: After evidence application, the compliance rate for audit criterion 1 increased from 0% at baseline audit to 70% at follow-up audit. The compliance rate for audit criterion 2 increased from 0% to 100%. The compliance rate for audit criterion 3 increased from 22 to 62%. The compliance rate for audit criterion 4 increased from 88 to 100%. The compliance rate for audit criterion 5 was 100% at baseline audit and follow-up audit. The compliance rate for audit criterion 6 increased from 0 to 55%. CONCLUSION: Implementation of the best evidence for the assessment and management of trismus of patients with NPC after radiotherapy is conducive to improving the compliance of clinical practice with the best evidence, standardizing clinical nursing practice, improving the quality of clinical nursing, and better preventing severe trismus in patients with NPC after radiotherapy.

Inhibitory Effect of Bovine Adipose-Derived Mesenchymal Stem Cells on Lipopolysaccharide Induced Inflammation of Endometrial Epithelial Cells in Dairy Cows.

Endometritis is a disease that affects reproductive health in dairy cows and causes serious economic damage to the dairy industry world-wide. Although in recent years, the application of mesenchymal stem cell (MSC) therapy for the treatment of inflammatory diseases has attracted much attention, there are few reports of the use of MSCs in dairy cows. In the present study, our objective was to explore the inhibitory effects of bovine adipose-derived mesenchymal stem cells (bAD-MSCs) on lipopolysaccharide (LPS) induced inflammation in bovine endometrial epithelial cells (bEECs) along with the potential underlying molecular mechanisms. We characterized isolated bAD-MSCs using cell surface marker staining and adipogenic/osteogenic differentiation, and analyzed them using immunofluorescence, flow cytometry (surface marker staining), and adipogenic and osteogenic differentiation. Furthermore, to understand the anti-inflammatory effects of bAD-MSCs on LPS induced bEEC inflammation, we used a bAD-MSC/bEEC co-culture system. The results showed that bAD-MSC treatments could significantly decrease LPS induced bEEC apoptosis and pro-inflammatory cytokine expression levels, such as interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Furthermore, our results showed that bAD-MSC treatments could also significantly downregulate LPS induced p38, IkB-a, and JAK1 phosphorylation and Bax protein expression levels, which are closely related to inflammatory progress and cellular apoptosis in bEECs. Our findings demonstrate that bAD-MSCs play an inhibitory role in LPS induced bEEC inflammation and provide new insights for the clinical therapy of endometritis in dairy cows.

G-CSF and IL-6 may be involved in formation of endometriosis lesions by increasing the expression of angiogenic factors in neutrophils.

Evidence accumulated in recent years has revealed that neutrophils are involved in the initial establishment of endometriosis, which is well-known as a chronic inflammatory disease. So far, why and how neutrophils promote the formation of early endometriosis are still unclear. In this study, using a mouse model of endometriosis, we demonstrated that endometriosis mice (EMs mice) had a significantly increased number of neutrophils in peritoneal fluids and lesions, and increased levels of granulocyte colony-stimulating factor (G-CSF) and IL-6 in serum and peritoneal fluids compared to the control group. In the neutrophils and uterine fragments co-injection experiment, neutrophils regulated by G-CSF and IL-6 had a similar effect to neutrophils from EMs mice, increasing the number, area, weight and microvessel density (MVD) of endometriotic lesions. Blocking the effect of G-CSF and IL-6 in EMs mice resulted in a decrease in the number, area and weight of endometriotic lesions. Following the depletion of neutrophils in vivo using a anti-Ly6G antibody, the MVD in the lesions of mice treated with neutrophils from EMs mice and neutrophils from pG/pI6 mice were significantly reduced. Neutrophils from EMs mice and neutrophils from pG/pI6 mice altered the expression levels of Mmp9, Bv8 and Trail genes compared to the neutrophils from PBS-treated mice. IL-6 together with G-CSF induced a higher expression of phospho-STAT3 and STAT3 in neutrophils. These findings suggest that neutrophils modulated by G-CSF and IL-6 through the STAT3 pathway alter the expression levels of the angiogenesis-related genes Mmp9, Bv8 and Trail, and may promote the establishment of early endometriosis.

Cosmc transfection decreases malignant behavior of Tn+ cells and enhances sensitivity to apoptosis when induced by Apo2L/TRAIL via alteration of O-glycan structure.

Cosmc mutations may cause abnormal O-glycosylation and result in Tn antigen expression. In the current study, it was discovered that proliferation and migration of Tn+ cells (Jurkat T and LS174T-Tn+ cells) with mutant Cosmc decreased after transfected Cosmc, and their sensitivity to apoptosis induced by Apo2L/TRAIL increased. Core 1-, 2-, and 3-derived O-glycans were absent in Tn+ cells. After Cosmc transfection, normal extended core 1-derived O-glycans appeared and were accompanied by increased T-synthase activity. Core 2-derived O-glycans appeared in transfected LS174T-Tn+ cells, and their structural types and levels were lower than those in LS174T-Tn- cells. Core 3-derived O-glycans were present only in LS174T-Tn- cells. The activity of C3GnT in LS174T-Tn+ cells was lower than that in LS174T-Tn- cells, and it was absent in Jurkat T cells. Cosmc transfection did not alter C3GnT activity or core 3-derived O-glycans in Jurkat T and LS174T-Tn+ cells. The results demonstrated that the composition and structure of O-glycans were different among various Tn+ cells, which not only affected cell malignant behavior but also modulated sensitivity to apoptotic stimuli. Thus, Cosmc transfection may effectively decrease the malignant behavior of Tn+ tumor cells and enhance their sensitivity to apoptosis when induced by Apo2L/TRAIL through modification of O-glycans.

Interleukin-37b inhibits the growth of murine endometriosis-like lesions by regulating proliferation, invasion, angiogenesis and inflammation.

Endometriosis is a gynecological disease with abnormal expression of interleukin (IL)-37 which can suppress inflammation and the immune system. Here we investigated the role of the IL-37b splice variant in endometriosis in vivo and in vitro. In a murine model of endometriosis, in vivo administration of IL-37b significantly inhibited the development of lesions judged by the number (P = 0.0213), size (P = 0.0130) and weight (P = 0.0152) of lesions. IL-37b had no effect on the early stage of lesion formation, however administration in the growth stage of lesions decreased the number (P = 0.0158), size (P = 0.0158) and weight (P = 0.0258) of lesions compared with PBS control, an effect that was not reversed by macrophage depletion. Expressions of inflammatory factors, matrix metalloproteinases and vascular endothelial growth factor-A mRNA/protein were significantly inhibited in ectopic lesions following IL-37b administration, and in uterine segments treated in vitro. In vitro treatment of uterine segments with IL-37b inhibited phosphorylation of Akt and Erk1/2 in uterine segments. Isolated mouse endometrial stromal treated with IL-37b and transfected with pIL-37b plasmid got suppressed cell proliferation, invasion, angiogenesis and the expression of inflammatory factors. In addition, transfection with pIL-37b significantly decreased the phosphorylation of Akt and Erk1/2. IL-37b also inhibited proliferation and the expression of inflammatory and angiogenesis factors in epithelial cell line RL95-2. These findings suggest that IL-37b may inhibit the growth of lesions by regulating proliferation, invasion, angiogenesis and inflammation through Akt and Erk1/2 signaling pathway.

Thalassorhabdomicrobium marinisediminis gen. nov., sp. nov., a member of the family Hyphomonadaceae isolated from the Bohai Sea.

A novel Gram-stain-negative bacterium, designated strain BH-SD16T, was isolated from a marine sediment sample collected in the Bohai Sea. Cells of strain BH-SD16T are aerobic, non-flagellated oval-shaped rods, showing oxidase- and catalase-positive activities. Growth occurs between 15-45 °C (optimum, 30 °C), at pH 6.0-9.0 (pH 7.0-7.5) and with 1-10 % (w/v) NaCl (3.0 %). Strain BH-SD16T contains C18 : 1ω7c (49.2 %), C16 : 0 (17.7 %) and C18 : 1ω7c 11-methyl (16.6 %) as the predominant fatty acids and ubiquinone-10 as the major respiratory quinone. The major polar lipids comprise phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, diphosphatidylglycerol and two glycolipids. The size of the draft genome is 3 442 538 bp, including 3213 protein-coding genes, 40 tRNA genes and three rRNA genes, and the DNA G+C content is 63.4 mol%. Strain BH-SD16T shows the highest 16S rRNA gene sequence similarity to Pseudooctadecabacter jejudonensis (95.7 %), strains of the genus Octadecabacter(95.4-95.6 %) and strains of the genus Loktanella(93.8-95.4 %). Phylogenetic trees based on 16S rRNA gene sequences show that strain BH-SD16T forms a distinct lineage within the family Hyphomonadaceae, which is also confirmed in the multigenic phylogenetic tree calculated by RAxML. Based on the results of phenotypic, chemotaxonomic and phylogenetic analysis, strain BH-SD16T is considered to represent a novel genus and species in the family Hyphomonadaceae, for which the name Thalassorhabdomicrobium marinisediminis gen. nov., sp. nov. is proposed. The type strain is BH-SD16T (=CCTCC AB 2017073T=KCTC 62201T).

Atom-Thick Interlayer Made of CVD-Grown Graphene Film on Separator for Advanced Lithium-Sulfur Batteries.

Lithium-sulfur batteries are widely seen as a promising next-generation energy-storage system owing to their ultrahigh energy density. Although extensive research efforts have tackled poor cycling performance and self-discharge, battery stability has been improved at the expense of energy density. We have developed an interlayer consisting of two-layer chemical vapor deposition (CVD)-grown graphene supported by a conventional polypropylene (PP) separator. Unlike interlayers made of discrete nano-/microstructures that increase the thickness and weight of the separator, the CVD-graphene is an intact film with an area of 5 × 60 cm2 and has a thickness of ∼0.6 nm and areal density of ∼0.15 µg cm-2, which are negligible to those of the PP separator. The CVD-graphene on PP separator is the thinnest and lightest interlayer to date and is able to suppress the shuttling of polysulfides and enhance the utilization of sulfur, leading to concurrently improved specific capacity, rate capability, and cycle stability and suppressed self-discharge when assembled with cathodes consisting of different sulfur/carbon composites and electrolytes either with or without LiNO3 additive.

CD3/CD28 dynabeads induce expression of tn antigen in human t cells accompanied by hypermethylation of the cosmc promoter.

Glycosylation is an important protein post-translational modification. In this process, the intermediate product, Tn antigen, arises from somatic mutations in core1ß3-galactosyltransferase-specific molecular chaperone (Cosmc), which is required for the formation of active core1ß3-galactosyltransferase (T-synthase). As a type of tumor-associated carbohydrate antigen, Tn antigen is mainly expressed in many human tumor cells and is absent in normal cells. Surprisingly, it is also expressed in normal activated T cells after in vitro stimulation, but the mechanism underlying its expression remains unclear. This study demonstrated that Tn antigen was expressed in activated T cells and that the percentage of positive (Tn+) cells increased and subsequently decreased within 72h after stimulation with CD3/CD28 Dynabeads, with peak expression occurring at 48h. During activation, interleukin-4 (IL-4) expression in the T-cell supernatant consistently increased with Tn+ cells, and was inversely correlated with serum interferon gamma (IFN-γ) levels. Compared with unactivated (without CD3/CD28 Dynabead stimulation) T cells, the level of T-synthase transcription in activated T cells did not significantly change, whereas T-synthase activity and Cosmc transcription significantly decreased, accompanied by a further increase in methylation of the Cosmc promoter. The results also showed that Cosmc transcription and translation decreased and then increased, and that Cosmc promoter methylation was a dynamic process during T cell activation. These data suggest that hypermethylation of the Cosmc promoter may induce the expression of Tn antigen in activated T cells.

Functional study of gene hp0169 in Helicobacter pylori pathogenesis.

Many virulence genes have been reported to play important roles in Helicobacter pylori pathogenesis. However the detailed mechanisms of many of them have not been completely clear. In this study, we found gene hp0169, encoding a putative collagenase (HpPrtC), was involved in pathogenesis of H. pylori. Recombinant HpPrtC shows activities to both native and heat-denatured collagens. This result indicated that HpPrtC may act as a virulence factor to help the bacterium colonize in their host stomach by degrading surrounding collagens. hp0169 was deleted by homologous recombination to study its function in bacterium-host cell interaction. For the pathogenic functions on the host cells, the hp0169 mutant exhibits no significant changes on inducing apoptosis of GES-1 cells. However, the viability and proliferation rate of GES-1 cells infected with mutant strain were higher than the cells infected with wild-type strain. These results indicated that except for its collagenolytic activity, HpPrtC might participate in H. pylori pathogenesis through an additional pathway. Functional studies on hp0169 involved in pathogenesis would shed light on deep understanding of the pathogenic mechanism of H. pylori.

Aberrant Cosmc genes result in Tn antigen expression in human colorectal carcinoma cell line HT-29.

The Tn antigen, which arises from mutation in the Cosmc gene is one of the most common tumor associated carbohydrate antigens. Cosmc resides in X24 encoded by a single gene and functions as a specific molecular chaperone for T-synthase. While the Tn antigen cannot be detected in normal cells, Cosmc mutations inactivate T-synthase and consequently result in Tn antigen expression within certain cancers. In addition to this Cosmc mutation-induced expression, the Tn antigen is also expressed in such cell lines as Jurkat T, LSC and LS174T. Whether the Cosmc mutation is present in the colon cancer cell line HT-29 is still unclear. Here, we isolate HT-29-Tn+ cells from HT-29 cells derived from a female colon cancer patient. These HT-29-Tn+ cells show a loss of the Cosmc gene coding sequence (CDS) leading to an absence of T-synthase activity and Tn antigen expression. Additionally, almost no methylation of Cosmc CpG islands was detected in HT-29-Tn+ as well as in HT-29-Tn- and Tn- tumor cells from male patients. In contrast, the methylation frequency of CpG island of Cosmc in normal female cells was ~50%. Only one active allele of Cosmc existed in HT-29-Tn+ and HT-29-Tn- cells as based upon detection of SNP sites. These results indicate that Tn antigens expression and T-synthase inactivity in HT-29-Tn+ cells can be related to the absence of CDS in Cosmc active alleles, while an inactive allele deletion of Cosmc in HT-29 cells has no influence on Cosmc function.