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Pseudomonas aeruginosa rugose small-colony variants (RSCVs) are frequently isolated from chronic infections, yet they are rarely reported in environmental isolates. Here, during comparative genomic analysis of two P. aeruginosa strains isolated from crude oil, we discovered a spontaneous in-frame deletion, wspAΔ280-307 , which led to hyper-biofilm and RSCV phenotypes. WspA is a homolog of methyl-accepting chemotaxis proteins (MCPs) that senses surfaces to regulate biofilm formation by stimulating cyclic-di-guanosine monophosphate (c-di-GMP) synthesis through the Wsp system. However, the methylation sites of WspA have never been identified. In this study, we identified E280 and E294 of WspA as methylation sites. The wspAΔ280-307 mutation enabled the Wsp system to lock into a constitutively active state that is independent of regulation by methylation. The result is an enhanced production of c-di-GMP. Sequence alignment revealed three conserved repeat sequences within the amino acid residues 280 to 313 (aa280-313) region of WspA homologues, suggesting that a spontaneous deletion within this DNA encoding region was likely a result of intragenic recombination and that similar mutations might occur in several related bacterial genera. Our results provide a plausible explanation for the selection of RSCVs and a mechanism to confer a competitive advantage for P. aeruginosa in a crude-oil environment. This article is protected by copyright. All rights reserved.

Background. Ulcerative colitis (UC) is a chronic inflammatory disease with increasing prevalence worldwide. Barrier defect in intestinal epithelial cells (IECs) is one of the main pathogeneses in UC. Pyroptosis is a programmed lytic cell death and is triggered by inflammatory caspases, while little is known about its role in UC. Methods. Differentially expressed genes (DEGs) were identified by comparing UC patients with healthy controls from the GEO datasets. The candidate genes involved in pyroptosis were obtained, and the underlying molecular mechanism in the progression of UC was explored in vivo and in vitro. Results. Pleckstrin homology domain leucine-rich repeat protein phosphatase 2 (PHLPP2), a protein phosphatase, was downregulated and involved in regulating inflammation-induced IEC pyroptosis by modulating the NF-κB signaling in UC through bioinformatics analysis. Moreover, we demonstrated that PHLPP2 was downregulated in UC patients and UC mice. Besides, we found that PHLPP2 depletion activated the NF-κB signaling and increased the expressions of caspase-1 P20, Gasdermin N, IL-18, and IL-1β contributing to IEC pyroptosis and inflammation in UC mice. Furthermore, we found that PHLPP2-/- mice developed hypersensitivity to dextran sulfate sodium (DSS) treatment toward colitis showing activated NF-κB signaling and dramatically induced expressions of caspase-1 P20, Gasdermin N, IL-18, and IL-1β. Mechanically, this inflammation-induced downregulation of PHLPP2 was alleviated by an NF-κB signaling inhibitor in intestinal organoids of PHLPP2-/- mice and fetal colonic cells. Conclusions. PHLPP2 downexpression activated the NF-κB signaling and promoted the IEC pyroptosis, leading to UC progression. Therefore, PHLPP2 might be an attractive candidate therapeutic target for UC.

Synaptotagmins (SYTs), constitute a family of 17 membrane-trafficking protein, palying crucial roles in the development and progression of human cancers. However, only very few studies have investigated the expression profile and prognostic values of SYTs family members in gastric cancer (GC). Therefore, we comprehensively evaluated the expression, methylation, prognosis and immune significance of SYTs family members through bioinformatics analysis from the online databases in GC. The expressions of SYT4, SYT9, and SYT14 were up-regulated, and negatively associated with their methylation levels in GC. Both the over-expression of SYT4, SYT9 and SYT14 and their hypomethylation levels contributed to an unsatisfactory overall survival (OS) and progression-free survival (PFS) in GC. Moreover, the low expressions of several methylation cg sites (cg02795029, cg07581146, cg15149095, cg19922137, cg25371503, cg26158959, cg02269161, cg03226737, cg08185661, cg16437728, cg22723056 and cg24678137) were significantly correlated with an unfavorable OS and PFS in GC. Furthermore, the expression of SYT4, SYT9 and SYT14 played a pivotal role in immune cells infiltration in GC. Collectively, our current finding suggested that SYT4, SYT9 and SYT14 might be potent prognostic indictors and promising immunotherapeutic targets for GC patients.

Strain YIM 132242T, isolated from lichen collected from Pu’er, Yunnan Province, China, was short-rod-shaped, Gram-reaction-negative, aerobic, catalase- and oxidase-positive. Growth of the strain was occurred at 10–39 °C (optimum, 28–35 °C), at pH 4.0–10.0 (optimum, pH 7.0–8.0) and at salinities of 0–8% (w/v) NaCl (optimum, 0–2%). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM 132242T belonged to the genus Paracoccus and had the highest levels of sequence similarity to Paracoccus aerius KCTC 42845T (97.0% similarity), Paracoccus sediminis CMB17T (96.8% similarity), and Paracoccus fontiphilus MVW-1T (96.4% similarity). The major fatty acid was identified as C18:1 ω7c (77.6%). The predominant respiratory quinone was ubiquinone-10 (Q-10). Polar lipid analysis indicated the presence of phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylcholine (PC), diphosphatidylglycerol (DPG), an unidentified lipid (L), and three unidentified phospholipids (PL1-PL3). Based on the draft genome sequence, the DNA G + C content of the strain was 67.1 mol%, and the values of average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) of strain YIM 132242T with Paracoccus aerius KCTC 42845T were 85.4% and 29.1%, respectively. On the basis of the data from this polyphasic characterization, strain YIM 132242T represents a novel species of the genus Paracoccus, for which the name Paracoccus lichenicola sp. nov. is proposed. The type strain is YIM 132242T (= KCTC 72463T = CGMCC1.17191T).

A Gram-stain-positive, non-motile, aerobic, rod-shaped, golden yellow-pigmented actinobacterium, designated strain YIM 123512T, was isolated from soil sampled at Gaoligong Mountain, Yunnan Province, PR China. The strain grew at 10–40 °C (optimum, 28 °C), with 0–3.0 % (w/v) NaCl (optimum, 0–1.0 %) and at pH 5.0–11.0 (optimum, pH 7.0). The major menaquinone was MK-8(H4) and the major cellular fatty acids (>10 %) were identified as iso-C16 : 0 and C17 : 1 ω8c. Strain YIM 123512T had ll-2, 6-diaminopimelic acid as the diagnostic cell-wall diamino acid. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, two unknown phospholipids and one unknown lipid. The genomic DNA G+C content of strain YIM 123512T was 72.7 mol% based on its draft genome sequence. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 123512T appeared to be most closely related to Nocardioides halotolerans KCTC 19274T (97.7 % sequence similarity) and to belong to the genus Nocardioides . The sequence similarity values of strain YIM 123512T to other currently described type strains of the genus Nocardioides were less than 97.0 %. Based on the draft genome sequence, the novel strain showed an average nucleotide identity value of 80.2 % and a digital DNA–DNA hybridization value of 23.1 % with the reference strain N. halotolerans KCTC 19274T. The results of the polyphasic taxonomic study including phenotypic, chemotaxonomic and phylogenetic analyses show that strain YIM 123512T represents a novel species of the genus Nocardioides , for which the name Nocardioides flavescens sp. nov. is proposed. The type strain is YIM 123512T (=KCTC 49303T=CGMCC 4.7628T).

A new diarylheptanoid, (1 R,2S,3S,5S)-2,3-dihydroxy-3',3''-dimethoxy-4'-de-O-methylcentrolobine (1) and a new bisabolane-type sesquiterpenoid, (1 R,7S)-1,12,13-trihydroxybisabola-3,10-diene (2), together with nineteen known compounds (3-21) were isolated from the EtOH extract of the stems and branches of Viscum coloratum (Kom.) Nakai. Their structures were elucidated by extensive analysis of 1 D and 2 D NMR spectra and from the HRESIMS. All the compounds were evaluated for their cytotoxic activity against eight human tumor cell lines.

A novel Gram-stain-positive, aerobic, non-spore-forming, irregular short rod-shaped actinobacterial strain, designated YIM 102482-1T, was isolated from the faeces of Macaca mulatta. Strain YIM 102482-1T grew optimally at 30–37 °C, at pH 8.0 and in the presence of 1.0–3.0% (w/v) NaCl. Similarly, analysis based on 16S rRNA gene sequences showed that strain YIM 102482-1T was a member of the genus Gulosibacter and most closely related to Gulosibacter feacalis NBRC 15706T (97.6 %), Gulosibacter bifidus NBRC 103089T (97.6 %), Gulosibacter chungangensis KCTC 13959T (96.4 %) and Gulosibacter molinativorax DSM 13485T (96.0 %), respectively. Furthermore, phylogenetic trees based on 16S rRNA gene sequences and genomic sequences demonstrated that strain YIM 102482-1T formed a distinct branch with all type strains of the genus Gulosibacter . The major whole-cell sugars and cellular fatty acids (>10.0 %) were ribose and rhamnose, and anteiso-C15 : 0, iso-C16 : 0 and C16 : 0, respectively. The predominant menaquinone was MK-9, and 2,4-diaminobutyric acid and ornithine were the diagnostic diamino acids in the cell-wall peptidoglycan. The dominant polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol and unidentified glycolipid. The DNA G+C content of YIM 102482-1T was 63.0 mol%. Based on analysis results of physiological, biochemical and chemotaxonomic data, strain YIM 102482-1T represents a novel species of the genus Gulosibacter , for which the name Gulosibacter macacae sp. nov. is proposed. The type strain is YIM 102482-1T(=DSM 102156T=CCTCC AB 2016023T).

Background: Microtubule Targeting Agents (MTAs) represent the most successful anticancer drugs for cancer chemotherapy. Through interfering with the tubulin polymerization and depolymerization dynamics, MTAs influence intracellular transport and cell signal pathways, inhibit cell mitosis and cell proliferation, and induce cell apoptosis and death. The tubulin maytansine site binding agents are natural or nature-derived products that represent one type of the MTAs that inhibit tubulin polymerization and exhibit potent antitumor activity both in vitro and in vivo. They are used as Antibody-Drug Conjugates (ADCs) in cancer chemotherapy. Methods: Using SciFinder® as a tool, the publications about maytansine, its derivatives, maytansine binding site, maytansine site binding agents and their applications as MTAs for cancer therapy were surveyed with an exclusion on those published as patents. The latest progresses in clinical trials were obtained from the clinical trial web. Results: This article presents an introduction about MTAs, maytansine, maytansine binding site and its ligands, the applications of these ligands as MTAs and ADCs in cancer therapy. Conclusion: The maytansine site binding agents are powerful MTAs for cancer chemotherapy. The maytansine site ligands-based ADCs are used in clinic or under clinical trials as cancer targeted therapy to improve their selectivity and to reduce their side effects. Further improvements in the delivery efficiency of the ADCs will benefit the patients in cancer targeted therapy.

Strain YIM 102744-1T was isolated from Rhinopithecus roxellanae fecal sample collected at Yunnan Wild Animal Park, Yunnan province, PR China. Cells were Gram-stain-negative, aerobic, non-motile and irregular rods. Optimal growth occurred in the presence of 0–1.0% (w/v) NaCl, at pH 7.0–8.0, and at 30 °C. The predominant ubiquinone was Q-10. The major cellular fatty acids (> 10.0%) were C18:1ω7c and C16:1ω7c/C16:1ω6c. The dominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The DNA G + C content was 62.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that YIM 102744-1T belonged to the family Rhodobacteraceae and shared the highest similarity with the type strain Gemmobacter intermedius 119/4T (96.6%). In addition, phylogenetic trees indicated that strain YIM 102744-1T formed a distinct clade together with the closest relative G. intermedius 119/4T. Based on the results of polyphasic taxonomic analysis, strain YIM 102744-1T is considered to represent a novel species within a new genus Falsigemmobacter, for which the name Falsigemmobacter faecalis gen. nov., sp. nov. is proposed. The type strain of Falsigemmobacter faecalis is YIM 102744-1T(= KCTC 52106T = CCTCC AB 2016031T). Because Gemmobacter intermedius 119/4T formed a branch with YIM 102744-1 in the phylogenetic trees and was clearly separated from the other members within the genus Gemmobacter, it is also proposed to transfer into the genus Falsigemmobacter as Falsigemmobacter intermedius comb. nov. (type strain 119/4T = CIP 110795T = LMG 28215T = CCM 8510T). The type species of the genus Falsigemmobacter is Falsigemmobacter intermedius gen. nov., comb. nov.

RNase J is a prokaryotic 5′-3′ exo/endoribonuclease that functions in mRNA decay and rRNA maturation. Here, we report a novel duplex unwinding activity of mpy-RNase J, an archaeal RNase J from Methanolobus psychrophilus, which enables it to degrade duplex RNAs with hairpins up to 40 bp when linking a 5′ single-stranded overhangs of ≥ 7 nt, corresponding to the RNA channel length. A 6-nt RNA-mpy-RNase J-S247A structure reveals the RNA-interacting residues and a steric barrier at the RNA channel entrance comprising two archaeal loops and two helices. Mutagenesis of the residues key to either exoribonucleolysis or RNA translocation diminished the duplex unwinding activity. Substitution of the residues in the steric barrier yielded stalled degradation intermediates at the duplex RNA regions. Thus, an exoribonucleolysis-driven and steric occlusion-based duplex unwinding mechanism was identified. The duplex unwinding activity confers mpy-RNase J the capability of degrading highly structured RNAs, including the bacterial REP RNA, and archaeal mRNAs, rRNAs, tRNAs, SRPs, RNase P and CD-box RNAs, providing an indicative of the potential key roles of mpy-RNase J in pleiotropic RNA metabolisms. Hydrolysis-coupled duplex unwinding activity was also detected in a bacterial RNase J, which may use a shared but slightly different unwinding mechanism from archaeal RNase Js, indicating that duplex unwinding is a common property of the prokaryotic RNase Js.

A novel actinobacterium, YIM 132087T, isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China. Cells are Gram-stain-positive, catalase-positive and oxidase-negative, aerobic, non-motile and short rod-shaped. Colonies are asporogenous, circular and white brown in colour. Optimal growth occured at 15−35 °C (optimum 28 °C), at pH 5.0−9.0 (optimum pH 6.0), and in the presence of 3% NaCl (w/v). The DNA G+C content of strain YIM 132087T based on the draft genome sequence was 71.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain YIM 132087T belonged to the genus Nakamurella and exhibited high levels of 16S rRNA gene sequence similarity with Nakamurella endophytica CGMCC 4.7038T (97.9%) and Nakamurella intestinalis NBRC 111844T (97.2%). The DNA–DNA hybridization values between strain YIM 132087T and its closest relatives are lower than 26%. Strain YIM 132087T had meso-diaminopimelic acid as the diagnostic cell-wall diamino acid, and MK-8(H4) as the predominant menaquinone. Predominant cellular fatty acids (> 10%) were iso-C16:0, iso-C15:0, C16:0 and anteiso-C15:0. The polar lipid profile were found to be diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, three unknown phospholipids, one unknown aminophospholipid and one unknown lipid. Based on phenotypic, phylogenetic and chemotaxonomic analysis, strain YIM 132087T belongs to the genus Nakamurella and represents a novel species of the genus Nakamurella, for which the name Nakamurella albus sp. nov., with type strain YIM 132087T (=CGMCC 4.7629T =NBRC 114017T), is proposed.

A Gram-stain-positive, yellow-pigmented, catalase-positive and oxidase-negative, strictly aerobic actinobacterium, designated strain YIM 131853T, was isolated from lichen collected from the South Bank of the Baltic Sea. The novel strain was non-spore-forming, short rod-shaped and motile with a single polar flagellum. The strain could grow at 4–37 °C (optimum, 28 °C), at pH 4.0–12.0 (pH 6.0) and at 0–3 % (w/v) NaCl (1 %). The DNA G+C content of strain YIM 131853T based on the draft genome sequence was 68.3 mol%. Predominant cellular fatty acids (>10 %) were identified as anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The polar lipid profile included diphosphatidylglycerol, dimannosyldiacylglycerol, three unknown glycolipids, two unknown phospholipids and one unknown lipid. Strain YIM 131853T had 2,4-diaminobutyric acid as the diagnostic cell-wall diamino acid, galactose and glucose as whole-cell sugars, and MK-10, MK-14, MK-13 and MK-12 as the major menaquinones. Although strain YIM 131853T exhibited a highest 16S rRNA gene sequence similarity (96.6 %) to Amnibacterium kyonggiense NBRC 109360T, phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain formed a tight lineage with Naasia aerilata NBRC 108725T (96.5 % 16S rRNA gene sequence similarity), which was the only species of genus Naasia . Based on the results of phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM 131853T should belong to the genus Naasia and represents a novel species of the genus Naasia , for which the name Naasia lichenicola sp. nov. is proposed. The type strain is YIM 131853T (=CGMCC 4.7565T=NBRC 113605T).

Patients with temporal lobe epilepsy (TLE) frequently suffer from memory disorders, and the pathological changes show widespread regional impairments in the brain. In lithium-pilocarpine (LIP)-treated rats with TLE, an abnormal hippocampal microstructure and functional connectivity have been observed. However, changes in other brain regions are still unclear. In the present study, diffusion tensor imaging and functional magnetic resonance imaging (MRI) signals were collected in LIP-TLE rats and controls using a 7.0 T MRI. Microstructural parameters and functional connectivity were calculated among regions of interest (ROIs), including the bilateral prefrontal cortex, amygdala, hippocampus and entorhinal cortex. A correlation analysis was further performed between the neuroimaging results and the behavioral performance in the novel object and novel location memory tests. In our results, TLE rats showed increased fractional anisotropy (FA) values in the hippocampus and decreased FA values in the amygdala and entorhinal cortex. In addition, decreased functional connectivity between the amygdala and the CA3, and increased connectivity between the prefrontal cortex and the CA1 were observed in the TLE rats compared to control rats. Moreover, FA values in the amygdala, the hippocampus and the entorhinal cortex, as well as the amygdala-CA3 and the prefrontal-CA1 connectivity correlated with the memory performance. Based on our results, both the microstructure and functional connections were impaired in memory-related brain regions of LIP-TLE rats. Furthermore, the abnormal changes in the microstructure and functional connectivity were related to behavioral deficits in object and location memory.

Background: The regulatory network of ulcerative colitis (UC)-associated miRNAs is not fully understood. In this study, we aim to investigate the global profile and regulatory network of UC associated miRNAs in the context of dextran sulfate sodium (DSS). Methods: UC was induced in C57BL mice using DSS. Differentially expressed miRNAs were screened by RNA sequencing and subjected to the Kyoto Encyclopedia of Genes and Genomes Pathway enrichment analysis. RT-qPCR was used to verify the differential expression of miRNAs and candidate target mRNA. Luciferase reporter vector bearing a miRNA binding site was constructed to verify the binding site of the miRNA on mRNA. Results:A total of 95 miRNAs (31 were up-regulated and 64 were down regulated) differentially expressed in the colonic tissues of the UC mice. Among the differentially expressed miRNAs, IL-25 pathway genes were enriched. Subsequent RT-qPCR confirmed a decreased expression of IL-25 and a significant up regulation of IL-25 target miRNAs including mmu-miR-135b-5p, mmu-miR-7239-5p and mmu-miR-691 in UC mice. Conclusion: Using the luciferase assay, we verified the biding sites of mmu-miR-135b-5p and mmu-miR-691 to the IL-25 3ʹUTR. In conclusion, mmu-miR-135b-5p:IL-25 and mmu-miR-691:IL-25 interactions are important pathways that may exert a protective role in UC., Graphical abstract

A Gram-negative, motile, aerobic and coccoid rod-shaped bacterium, designated strain YIM132180T, was isolated from a Lepraria sp. lichen collected from Pu’er, Yunnan Province, China. The strain grew at 15–35 °C (optimum, 25–28 °C), at 0–2% (w/v) NaCl (optimum, 0–1%) and at pH 6.0–9.0 (optimum, pH 7.0). The 16S rRNA gene sequence showed that strain YIM132180T had highest similarity (96.4%) with Aureimonas endophytica 2T4P-2-4T, followed by Aureimonas ureilytica NBRC 106430T (95.7%) and Aureimonas rubiginis CC-CFT034T (95.6%). Phylogenetic analysis showed that the strain grouped with species of the genus Aureimonas. The genomic sequence was 4,779,519 bp and contained 4584 coding sequences (CDSs), 54 RNA genes, 3 complete rRNA genes and 47 tRNA genes. The major fatty acids (>10%) of strain YIM132180T were C18:1ω7c, C-16:0 and C19:0 cyclo ω8c. The predominant menaquinone was ubiquinone 10 (Q-10). The polar lipid profile comprised diphosphatidylglycerol, phosphatidylcholine, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, unidentified phospholipid, amino lipid, lipid and most importantly sulfoquinovosyldiacylglycerol (SQDG). Based on the draft genome sequence, the G +C content of strain YIM132180T was 68.4 mol%. The results of the polyphasic taxonomic study, including phenotypic, chemotaxonomic, and phylogenetic analyses, showed that strain YIM132180T represents a novel species of the genus Aureimonas, for which the name Aureimonas leprariae sp. nov. is proposed. The type strain is YIM 132180T (=KCTC 72462T = CGMCC 1.17389T).

A novel actinobacterium, YIM 132084T, was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China and identified by a polyphasic taxonomic approach. The strain was Gram-stain-positive, aerobic, catalase-positive, oxidase-negative, non-motile and coccus-shaped. Colonies were round, convex, smooth and light orange yellow in colour. It grew at 10–40 ℃ (optimum 28 ℃), at pH 6.0–11.0 (optimum pH 7.0) and in the presence of 0–4 % NaCl (optimum 0 %). Strain YIM 132084T comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol as the major polar lipids, MK-8(H4) as the predominant menaquinone, and anteiso-C15:0, anteiso-C17:0, iso-C15:0 and iso-C16:0 as major fatty acids. Strain YIM 132084T had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The 16S rRNA gene sequence showed high level of similarity with Nakamurella flavida KCTC 19127T (97. 7%) and Nakamurella flava CGMCC 4.7524T (97.7%). The G + C content of the genomic DNA was 72.4 mol%. Strain YIM 132084T showed an average nucleotide identity value of 76.1 % and 74.9 %, a digital DNA-DNA hybridizations value of 20.9 % and 20.6 % with the reference strains Nakamurella flavida and Nakamurella flava based on draft genome sequences, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain YIM 132084T represents a novel species of the genus Nakamurella, for which the name Nakamurella leprariae sp. nov. is proposed. The type strain is YIM 132084T (= CGMCC 4.7667T = NBRC 114280T = KCTC 49367T).

The leaf is an extremely important plant organ exhibiting a broad range of phenotypic variation. In watermelon (Citrullus lanatus), leaf spotting is a rare, valuable trait that can be used by breeders for selection at early growth stages. In this study, we tested a seven-generation family to determine the inheritance and genetic basis of this trait. As revealed by analysis of the lesion mimic mutant clalm, leaf spotting is controlled by a single dominant gene. Whole genome resequencing–bulked segregant analysis demonstrated that this gene is located on chromosome 4 from 3,760,000 bp to 7,440,000 bp, a region corresponding to a physical distance of 3.68 Mb encompassing approximately 72 annotated genes and eight non-synonymous coding SNPs. According to quantitative real-time PCR analysis, the expression level of ClCG04G001930 was significantly lower in the clalm mutant than in normal watermelon. The predicted target gene, ClCG04G001930, encodes a fatty acid amide hydrolase protein that regulates a variety of neurobehavioral processes in animals. Twelve-five SNPs were identified in the ClCG04G001930 gene of F2 individuals of the clalm mutant. RNA interference of the ClCG04G001930 gene, designated as ClPAD4, yielded transgenic lines whose leaves gradually developed chlorotic lesions over 3 weeks. Our results suggest that ClPAD4 is the gene responsible for leaf spotting in the clalm mutant. Our findings may serve as a foundation for elucidating the mechanism underlying the spotted leaf trait and should be useful for marker-assisted selection breeding in watermelon.

BackgroundColorectal cancer (CRC) is a common malignant tumor with high metastatic and recurrent rates. This study probes the effect and mechanism of long non-coding RNA MIR31HG on the progression of CRC cells.Materials and MethodsQuantitative real-time PCR (qRT-PCR) was used to analyze the expression of MIR31HG and miR-361-3p in CRC tissues and normal tissues. Gain- or loss-of-function assays were conducted to examine the roles of MIR31HG, miR-361-3p and YY1 transcription factor (YY1) in the CRC progression. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and colony formation experiment were conducted to test CRC cell proliferation. CRC cell invasion was determined by Transwell assay. The glucose detection kit and lactic acid detection kit were utilized to monitor the levels of glucose and lactate in CRC cells. The glycolysis level in CRC cells was examined by the glycolytic stress experiment. Western blot was performed to compare the expression of glycolysis-related proteins (PKM2, GLUT1 and HK2) and angiogenesis-related proteins (including VEGFA, ANGPT1, HIF1A and TIMP1) in HUVECs. The binding relationships between MIR31HG and miR-361-3p, miR-361-3p and YY1 were evaluated by the dual-luciferase reporter assay and RNA immunoprecipitation (RIP).ResultsMIR31HG was up-regulated in CRC tissues and was associated with poorer prognosis of CRC patients. The in-vitro and in-vivo experiments confirmed that overexpressing MIR31HG heightened the proliferation, growth, invasion, glycolysis and lung metastasis of CRC cells as well as the angiogenesis of HUVECs. In addition, MIR3HG overexpression promoted YY1 mRNA and protein level, and forced overexpression of YY1 enhanced MIR31HG level. Overexpressing YY1 reversed the tumor-suppressive effect mediated by MIR31HG knockdown. miR-361-3p, which was inhibited by MIR31HG overexpression, repressed the malignant behaviors of CRC cells. miR-361-3p-mediated anti-tumor effects were mostly reversed by upregulating MIR31HG. Further mechanism studies illustrated that miR-361-3p targeted and negatively regulated the expression of YY1.ConclusionThis study reveals that MIR31HG functions as an oncogenic gene in CRC via forming a positive feedback loop of MIR31HG-miR-361-3p-YY1.

The highly conserved multifunctional polymerase-associated factor 1 (Paf1) complex (PAF1C), composed of five core subunits Paf1, Leo1, Ctr9, Cdc73, and Rtf1, participates in all stages of transcription and is required for the Rad6/Bre1-mediated monoubiquitination of histone H2B (H2Bub). However, the molecular mechanisms underlying the contributions of the PAF1C subunits to H2Bub are not fully understood. Here, we report that Ctr9, acting as a hub, interacts with the carboxyl-terminal acidic tail of Rad6, which is required for PAF1C-induced stimulation of H2Bub. Importantly, we found that the Ras-like domain of Cdc73 has the potential to accelerate ubiquitin discharge from Rad6 and thus facilitates H2Bub, a process that might be conserved from yeast to humans. Moreover, we found that Rtf1 HMD stimulates H2Bub, probably through accelerating ubiquitin discharge from Rad6 alone or in cooperation with Cdc73 and Bre1, and that the Paf1/Leo1 heterodimer in PAF1C specifically recognizes the histone H3 tail of nucleosomal substrates, stimulating H2Bub. Collectively, our biochemical results indicate that intact PAF1C is required to efficiently stimulate Rad6/Bre1-mediated H2Bub.

A novel Gram-stain-negative strain, WQ 117T, isolated from the faeces of Rhinopithecus bieti collected at Yunnan Snub-nosed Monkey National Park, Yunnan province, PR China, was subjected to a polyphasic taxonomic study. The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate represented a member of the genus Faecalibacter , sharing 97.64 % sequence similarity with the type strain Faecalibacter macacae YIM 102668T. The G+C content of the genomic DNA of WQ117T was 30.5 mol%. The predominant isoprenoid quinone was MK-6. The major cellular fatty acids was iso-C15 : 0. The whole genome average nucleotide identity (gANI) values and the digital DNA–DNA hybridization values between WQ 117T and YIM 102668T were 79.66 % and 22.20 %, respectively. Growth occurred at 0–50 °C (optimally at 28–35 °C), pH 7.0–9.0 (optimally at pH 8.0) and with 0–2 % (w/v) NaCl (optimally without NaCl). On the basis of the taxonomic evidence, a novel species, Faecalibacter rhinopitheci sp. nov., is proposed. The type strain is WQ 117T (=KCTC 82394T=CCTCC AA 2020027T).

A novel Actinobacterium strain YIM 131861 T, was isolated from lichen collected from the South Bank Forest of the Baltic Sea, Germany. It was Gram-stain-positive, strictly aerobic, catalase positive and oxidase negative, yellow pigmented. Cells were motile with a polar flagellum, irregular rod shaped and did not display spore formation. The strain grew at 15 − 30 °C (optimum 25 °C), at pH 6.0 − 10.0 (optimum pH 7.0) and in the presence of 0 − 1.5% (w/v) NaCl (optimum 1%). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 131861 T belonged to the genus Glaciibacter, and exhibited a high sequence similarity (96.4%) with Glaciibacter superstes NBRC 104264 T. The genomic DNA G + C content of strain YIM 131861 T was 68.2 mol%. The average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values between strain YIM 131861 T and Glaciibacter superstes NBRC 104264 T were 73.2 and 19.9% based on the draft genome sequence. The cell-wall peptidoglycan type was B2γ and contained the 2, 4-diaminobutyric acid as the diagnostic amino acid. Whole cell sugars were galactose, rhamnose, ribose and glucose. It contained MK-12 and MK-13 as the predominant menaquinones. The major cellular fatty acids (> 10%) were identified as anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol and two unknown glycolipids. Based on the results of phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM 131861 T should belong to the genus Glaciibacter and represents a novel species of the genus Glaciibacter, for which the name Glaciibacter flavus sp. nov. is proposed. The type strain is YIM 131861 T (= CGMCC 1.16588 T = NBRC 113572 T).

Biofilms are microbial communities that are embedded in the extracellular matrix. The exopolysaccharide (EPS) is a key component of the biofilm matrix that maintains the structure of the biofilm and protects the bacteria from antimicrobials. Microbial glycoside hydrolases have been exploited to disrupt biofilms by breaking down EPSs. PssZ has recently been identified as a glycoside hydrolase that can disperse aggregates of Listeria monocytogenes. In this study, the crystal structure of PssZ has been determined at 1.6 Å resolution. PssZ belongs to glycoside hydrolase family 8 and adopts a classical (α/α)6-barrel fold. This architecture forms a deep groove which may serve as the substrate-binding pocket. The conserved catalytic residues (Glu72, Trp110, Asn119, Phe167, Tyr183 and Asp232) are localized at the centre of the groove. This crystal structure will help to improve the understanding of the hydrolytic mechanism of PssZ and its application as a biofilm disrupter.

Transmembrane chemoreceptors are widely present in Bacteria and Archaea. They play a critical role in sensing various signals outside and transmitting to the cell interior. Here, we report the structure of the periplasmic ligand-binding domain (LBD) of the transmembrane chemoreceptor MCP2201, which governs chemotaxis to citrate and other organic compounds in Comamonas testosteroni. The apo-form LBD crystal revealed a typical four-helix bundle homodimer, similar to previously well-studied chemoreceptors such as Tar and Tsr of Escherichia coli. However, the citrate-bound LBD revealed a four-helix bundle homotrimer that had not been observed in bacterial chemoreceptor LBDs. This homotrimer was further confirmed with size-exclusion chromatography, analytical ultracentrifugation and cross-linking experiments. The physiological importance of the homotrimer for chemotaxis was demonstrated with site-directed mutations of key amino acid residues in C. testosteroni mutants.

Salmonella is one of the most important foodborne pathogens associated with animal and human diseases. In this study, 672 samples of fresh meat (pork, 347; chicken, 196; and duck, 129) wer...

L,D-transpeptidases, widely distributed in bacteria and even in the difficult-to-treat ESKAPE pathogens, can confer antibacterial resistance against the traditional β-lactam antibiotics through bypass of the 4 → 3 transpeptide linkage. LdtMt2, a l , d -transpeptidase in Mycobacteria tuberculosis, is essential for bacterial virulence and is considered as a potential anti-tuberculosis target inhibited by carbapenems. Diverse interaction modes between carbapenems and LdtMt2 have been reported, there are only limited evidences to validate those interaction modes. Herein, we identified the stable binding states of two carbapenems, imipenem and ertapenem, via crystallographic and biochemical studies, discovered that they adopt similar binding conformations. We further demonstrate the absence of the 1-β-methyl group in imipenem and the presence of both Y308 and Y318 residues in LdtMt2 synergistically resulted in one order of magnitude higher affinity for imipenem than ertapenem. Our study provides a structural basis for the rational drug design and evolvement of novel carbapenems against bacterial L,D-transpeptidases.

Numerous studies have shown that the expression of circular RNA (circRNA) is closely related to the malignant progression of cancer. However, the role of circ-MFN2 in colorectal cancer (CRC) is unclear. Our study aims to explore the role and mechanism of circ-MFN2 in CRC progression. The relative expression levels of circ-MFN2, microRNA (miR)-574-3p and insulin-like growth factor 1 receptor (IGF1R) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was determined using 3-(4, 5-dimethyl-2 thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The colony number and radioresistance of cells were assessed using colony formation assay. Moreover, the migration and invasion of cells were measured using transwell assay. Tumor xenograft model was constructed to evaluate the effect of circ-MFN2 knockdown on CRC tumor growth. Furthermore, dual-luciferase reporter assay was used to verify the interaction between miR-574-3p and circ-MFN2 or IGF1R. In addition, the protein level of IGF1R was evaluated by western blot (WB) analysis. Circ-MFN2 expression was elevated in CRC tissues and cells. Knockdown of circ-MFN2 restrained the proliferation, migration, invasion, and radioresistance of CRC cells in vitro. Furthermore, silenced circ-MFN2 also reduced the tumor volume and weight of CRC in vivo. MiR-574-3p could be sponged by circ-MFN2, and its inhibitor reversed the suppression effect of circ-MFN2 silencing on CRC progression. Moreover, IGF1R was a target of miR-574-3p, and its overexpression reversed the inhibition effect of miR-574-3p mimic on CRC progression. In addition, circ-MFN2 could positively regulate IGF1R expression by sponging miR-574-3p. Our results revealed that circ-MFN2 promoted the proliferation, metastasis and radioresistance of CRC through regulating the miR-574-3p/IGF1R axis, suggesting that circ-MFN2 might be a novel therapeutic biomarker for CRC.

Background: Synaptotagmins (SYTs) family members, consisting of 17 isoforms, paly crucial roles in the development and progression of human cancer. However, there is few studies on the expression and prognostic values of SYTs family in gastric cancer (GC). Methods: We comprehensively evaluate the expression, methylation, prognosis and immune significance of SYTs family members through bioinformatics analysis from the online databases in GC. Results: The expressions of SYT4, SYT9 and SYT14 were up-regulated, and negatively associated with their methylation levels in GC. Both SYT4, SYT9 and SYT14 overexpression and their hypomethylation levels contributed to unsatisfactory overall survival (OS) and progression-free survival (PFS) in GC. Moreover, the low expression levels of methylation cg sites (cg02795029, cg07581146, cg15149095, cg19922137, cg25371503, cg26158959, cg02269161, cg03226737, cg08185661, cg16437728, cg22723056 and cg24678137) were significantly correlated with unfavorable OS and PFS in GC. Furthermore, the expression of SYT4, SYT9 and SYT14 played a pivotal role in immune cells infiltration in GC. Conclusion: SYT4, SYT9 and SYT14 might be potent prognostic indictors and promising immunotherapy targets for the patients with GC.

Abscisic acid (ABA) is an important phytohormone regulating plant growth, development and stress responses. A multitude of key factors implicated in ABA signaling have been identified; however, the regulation network of these factors needs for further information. AtS40.4, a plant-specific DUF584 domain-containing protein, was identified previously as a senescence regulator inArabidopsis. In this study, our finding showed that AtS40.4 was negatively involved in ABA signaling during seed germination and early seedling growth.AtS40.4was highly expressed in seeds and seedlings, and the expression level was promoted by ABA. AtS40.4 was localized both in the nucleus and the cytoplasm. Moreover, the subcellular localization pattern of AtS40.4 was affected by ABA. The knockdown mutants ofAtS40.4exhibited an increased sensitivity to ABA, whereas the overexpression ofAtS40.4decreased the ABA response during seed germination and seedling growth ofArabidopsis. Furthermore, AtS40.4 was involved in ABRE-dependent ABA signaling and influenced the expression levels ofABA INSENTIVE(ABI)1-5andSnRK2.6. Further genetic evidence demonstrated thatAtS40.4functioned upstream ofABI4.These findings support the notion that AtS40.4 is a novel negative regulator of the ABA response network during seed germination and early seedling growth.

A Gram-stain-positive, mycelium-forming actinobacterium, YIM 121974T was isolated from an extreme arid soil sample collected at Yuanmou Earth Forest, Yunnan Province, PR China. Classification using a polyphasic approach suggested that strain YIM 121974T belonged to the genus Glycomyces and was closely related to Glycomyces dulcitolivorans SJ-25T (98.3 %), Glycomyces scopariae YIM 56256T (98 %), Glycomyces mayteni YIM 61331T (97.9 %), Glycomyces albidus NEAU-7082T (97.9 %), Glycomyces sambucus CGMCC 4.3147T (97.7 %), Glycomyces artemisiae IXS4T (97.6 %) and Glycomyces parisis CPCC 204357T (97.5 %), but could be distinguished from its closest relatives by a combination of phenotypic and phylogenetic features. Average nucleotide identity values of YIM 121974T to its closest phylogenetic neighbours were 70.7–88.9 %, which are lower than the threshold of 95 %. The digital DNA–DNA hybridization values between YIM 121974T and these relative species were 18.0–36.3 %, which are also well below the cut-off value (>70 %) for species delineation. The DNA G+C content of strain YIM 121974T was 72.3 mol% (draft genome sequence). The predominant menaquinone was MK-11. The phospholipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, phosphoglycolipid and glycolipid. The major fatty acid compositions were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The draft genome of isolate YIM 121974T was found to contain 11 secondary metabolite biosynthesis gene clusters by using the antiSMASH server. Based on the above observations, strain YIM 121974T could be distinguished from closely related species belonging to the genus Glycomyces . Thus, strain YIM 121974T represents a novel species of the genus Glycomyces , for which the name Glycomyces terrestris sp. nov. is proposed. The type strain is YIM 121974T (=KCTC 39870T=DSM 106742T).

Heterotrophic nitrifiers are able to oxidize and remove ammonia from nitrogen-rich wastewaters but the genetic elements of heterotrophic ammonia oxidation are poorly understood. Here, we isolated and identified a novel heterotrophic nitrifier, Alcaligenes ammonioxydans sp. nov. strain HO-1, oxidizing ammonia to hydroxylamine and ending in the production of N2 gas. Genome analysis revealed that strain HO-1 encoded a complete denitrification pathway but lacks any genes coding for homologous to known ammonia monooxygenases or hydroxylamine oxidoreductases. Our results demonstrated strain HO-1 denitrified nitrite (not nitrate) to N2 and N2O at anaerobic and aerobic conditions respectively. Further experiments demonstrated that inhibition of aerobic denitrification did not stop ammonia oxidation and N2 production. A gene cluster (dnfT1RT2ABCD) was cloned from strain HO-1 and enabled E. coli accumulated hydroxylamine. Sub-cloning showed that genetic cluster dnfAB or dnfABC already enabled E. coli cells to produce hydroxylamine and further to 15N2 from (15NH4)2SO4. Transcriptome analysis revealed these three genes dnfA, dnfB and dnfC were significantly upregulated in response to ammonia stimulation. Taken together, we concluded that strain HO-1 has a novel dnf genetic cluster for ammonia oxidation and this dnf genetic cluster encoded a previously unknown pathway of direct ammonia oxidation (Dirammox) to N2.

Hydroxylamine is a key intermediate of microbial ammonia oxidation and plays an important role in the biogeochemical cycling of N-compounds. Hydroxylamine is oxidized to NO or N2O by hydroxylamine oxidases or cytochrome P460 from heterotrophic or autotrophic bacteria, but its enzymatic oxidation to N2 has not yet been observed. Here, we report on the discovery of a novel oxidase that converts hydroxylamine to N2 from the newly isolated heterotrophic nitrifier Alcaligenes strain HO-1. Strain HO-1 accumulated hydroxylamine and produced N2 from ammonia oxidation. Using transcriptome analysis and heterologous expression via fosmid library screening, we identified three genes (dnfABC) of strain HO-1 that enabled E. coli cells not only to produce hydroxylamine from 15N-labelled ammonium but also to further convert it to 15N2. The three genes were individually cloned and expressed, and their translational products DnfA, DnfB, and DnfC were purified. In vitro DnfA bound to hydroxylamine and catalyzed the conversion of hydroxylamine to N2 in the presence of FAD, NADH and O2. Thus, DnfA was identified as a novel hydroxylamine oxidase and catalyzed a previously unknown N-N bond forming reaction with a yet-to-be discovered mechanism. DnfA homologs were detected in different bacterial groups, suggesting that hydroxylamine oxidation to nitrogen might occur in additional microbial taxa.

De-feng Meng,1 Hua Shao,2 Chuan-bo Feng2 1Department of Oncology Surgery, North China University of Science and Technology Affiliated Hospital, Tangshan, Hebei, People’s Republic of China; 2Department of Thyroid and Breast Surgery, The Second People’s Hospital of Lianyungang City, Lianyungang, Jiangsu, People’s Republic of ChinaCorrespondence: Chuan-bo FengThe Second People’s Hospital of Lianyungang City, 41 Hailian East Road, Lianyungang City, Jiangsu, 222000, People’s Republic of ChinaTel +86-0518-85776181Email amdoreducn@163.comPurpose: Long non-coding RNAs (lncRNAs) are known to regulate tumorigenesis. Although breast cancer tissues show a high expression of LINC00894, its specific biological role in breast cancer progression is still unknown. In this study, lncRNA microarray was used to analyze the lncRNA expression in breast cancer tissues, and LINC00894 was selected for further analysis.Materials and Methods: Expression of LINC00894 in 45 pairs of breast cancer tissues and normal tissues obtained from patients with breast cancer was assessed by quantitative reverse transcription-PCR, while proliferation and invasion of breast cancer cells were assessed using a Cell Counting Kit-8 (CCK-8), EdU assay, colony formation experiment, and transwell assays. A dual-luciferase reporter gene assay and bioinformatics analysis were employed to detect potential targets of LINC00894. Additionally, RNA Binding Protein Immunoprecipitation (RIP) and Western blot assays were utilized to clarify its interaction and roles in the regulation of breast cancer progression.Results: High expression of LINC00894 was observed in breast cancer cells, and its overexpression significantly expedited cell proliferation and invasion. Moreover, LINC00894 positively regulated the expression of ZEB1 by competitively binding to miR-429.Conclusion: Taken together, these results suggest that LINC00894 competitively binds to miR-429 to mediate ZEB1 expression; consequently, it is implicated to play a role in the progression of breast cancer.Keywords: lncRNA, ceRNA, proliferation, invasion, breast cancer

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies worldwide due to its ineffective diagnosis and poor prognosis. It is essential to identify differentially expressed genes (DEGs) in PDAC to gain new insights into its underlying molecular mechanisms, as well as identify potential diagnostic and therapeutic targets. We screened 135 DEGs from the GSE15417, GSE16515, and GSE28735 PDAC and normal pancreatic tissue microarray data sets, and identified 16 DEGs that were correlated with PDAC prognosis through the Kaplan-Meier survival analysis and log-rank tests. The Cancer Genome Atlas and Oncomine databases validated the expression levels of 16 candidate genes (SLC6A14, GPRC5A, IFI27, ERP27, SDR16C5, SIDT2, TCN1, COL12A1, MMP1, CEACAM6, DKK1, ITGA2, KRT19, PLAU, ANO1, and GABRP). Weighted gene coexpression network analysis (WGCNA) and protein and protein interaction (PPI) analysis identified three hub genes-ERP27, ITGA2, and MMP1-that are likely important in PDAC prognosis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis demonstrated that they were enriched in functions of extracellular matrix organization, extracellular structure organization, and positive regulation of cell migration. Taken together, we identified three pivotal genes for PDAC, which can improve our understanding of its pathogenesis, progression, and prognosis.

Maturation arrest is a common cause of male infertility which has caused worldwide concern, and its pathophysiological process remains further elucidation. Our study aimed to identify genetic characteristics of maturation arrest by comparing gene expression between maturation arrest and normal samples using microarray technology. A total of 6,373 genes were identified differentially expressed (p .05, fold change 2.0 or-2.0) and 1,594 genes were selected as statistically significant after Bonferroni correction, including 419 up-regulated and 1,175 down-regulated genes. Microarray data were validated by quantitative reverse transcriptase-polymerase chain reaction. Bioinformation analysis was performed to explore genetic function of statistically significant genes. Gene Ontology results showed the statistically significant genes enriched in sexual reproduction, spermatogenesis and male gamete generation. Reactome pathway analysis highlighted the olfactory signalling pathway, fertilisation, developmental biology, etc. One module and eight hub genes were found to be involved in ubiquitin-mediated proteolysis and may affect as indicators of spermatogenic process through protein-protein interaction analysis. Our study provided a comprehensive genetic characteristic of differential expressed genes in testicular tissues from maturation arrest patients and speculated several genes as potential indicators of disease.

Objective The aim of this study is to investigate the role of close homolog of L1 (CHL1) on inflammatory bowel disease (IBD), and the correlation with the balance of Th17/Treg. Methods Dextran sodium sulfate (DSS)-induced IBD mice model was established. CHL1 knockout (KO) mice and CHL1 wild-type (WT) mice were subjected to DSS. CHL1 expression was detected using qRT-PCR. Weight was recorded daily, and disease activity index (DAI) score was assessed. The colon length and histological changes were measured. The number of neutrophils, macrophages and T cells was observed by immunohistochemistry. The expression of inflammatory cytokines and the proportion of Th17/Treg cells were detected by qRT-PCR and flow cytometry. The expression of RORγt, STAT3 and Foxp3 was detected by using immunohistochemistry and Western blot. Results CHL1 expression was upregulated in DSS-induced IBD mice. DSS-CHLl-KO mice exhibited less weight loss than the DSS-CHLl-WT mice. The DAI score and histological score were decreased in DSS-CHLl-KO mice compared with DSS-CHLl-WT mice, while colon length was increased. Number of neutrophils, macrophages and T cells, and expression of TNF-α, IL-6, IL-17A, IL-21 and IL-23 were decreased in DSS-CHLl-KO mice, while IL-10 expression was increased. Moreover, CHL1-deficient inhibited Th17 cells differentiation and promoted Treg cells differentiation in IBD mice. CHL1-deficient also inhibited the expression of RORγt and STAT3, and promoted the expression of Foxp3 in IBD mice. Conclusion CHL1-deficient reduces the inflammatory response by regulating the balance of Th17/Treg in mice with IBD. CHL1 is expected to be a new target for the treatment of IBD.

A novel bacterial strain, designated YIM 132548 T, was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China. The organism was Gram-stain negative, aerobic and methylotrophic. The cell was catalase positive and oxidase negative, asporogenous, rod-shaped and motile with three polar flagella. The strain could grow at 15–30 °C (optimum, 20 °C), at pH 6.0–9.0 (optimum, pH 7.0) and does not grow in the presence of NaCl. According to the 16S rRNA gene sequence analysis, strain YIM 132548 T showed high levels of 16S rRNA gene sequence similarity with Methylobacterium soli YIM 48816 T (97.6%) and Methylobacterium durans NBRC 112876 T (97.3%), less than 97.0% with other validly named type strains of the genus Methylobacterium. Ubiquinone Q-10 was the predominant respiratory ubiquinone. The predominant cellular fatty acid was identified as summed feature 8 (C18:1ω7c). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. The DNA G + C content of the draft genome sequence is 70.2 mol%. The average nucleotide identity and digital DNA–DNA hybridizations values of strain YIM 132548 T with M. soli YIM 48816 T and M. durans NBRC 112876 T were 87.0% and 82.0%, 40.6% and 27.2% based on draft genome sequences, respectively. On the basis of phylogenetic, chemotaxonomic, phenotypic and genomic data, strain YIM 132548 T is concluded to represent a novel species of the genus Methylobacterium, for which the name Methylobacterium planium sp. nov. is proposed. The type strain is YIM 132548 T (= CGMCC 1.17323 T = NBRC 114056 T).

Gastric cancer (GC) is one of the most common malignancies that threatens human health. As the molecular mechanisms unerlying GC are not completely understood, identification of genes related to GC could provide new insights into gene function as well as potential treatment targets. We discovered that UGT2B15 may contribute to the pathogenesis and progression of GC using GEO data and bioinformatic analysis. Using TCGA data, UGT2B15 mRNA was found to be significantly overexpressed in GC tissues; patients with higher UGT2B15 had a poorer prognosis. It was further discovered that UGT2B15 and FOXA1 were both upregulated, and UGT2B15 and Foxa1 were positively correlated in GC. It is known that Foxa1 is a vital threshold to activate the Hippo‑YAP signaling pathway. In addition, we suggest that a potential molecular mechanisms includes UGT2B15 which may upregulate Foxa1, activate the Hippo‑YAP signaling pathway and contribute to the development of GC. Taken together, our findings demonstrate that UGT2B15 may be an oncogene in GC and is a promising therapeutic target for cancer treatment.

Fifteen previously undescribed nor-clerodane diterpenoid glucosides tinosinesides C-Q (1-15), along with four known analogues (16-19), were isolated from the stems of Tinospora sinensis. The structures of the new compounds were elucidated by spectroscopic means, and their absolute configurations were established on the basis of time-dependent density functional theory (TD-DFT) based electronic circular dichroism (ECD) calculation and chemical methods. All the isolates were evaluated for their inhibitory effects on cystathionine γ-lyase (CSE), a natural enzyme responsible for the synthesis of H2S. Compounds 4 and 5 represent rare examples of natural CSE inhibitors and the possible binding mode to CSE was further probed by molecular docking experiment.

RNase J, a prokaryotic 5'-3' exo/endoribonuclease, contributes to mRNA decay, rRNA maturation and post-transcriptional regulation. Yet the processive-exoribonucleolysis mechanism remains obscure. Here, we solved the first RNA-free and RNA-bound structures of an archaeal RNase J, and through intensive biochemical studies provided detailed mechanistic insights into the catalysis and processivity. Distinct dimerization/tetramerization patterns were observed for archaeal and bacterial RNase Js, and unique archaeal Loops I and II were found involved in RNA interaction. A hydrogen-bond-network was identified for the first time that assists catalysis by facilitating efficient proton transfer in the catalytic center. A conserved 5'-monophosphate-binding pocket that coordinates the RNA 5'-end ensures the 5'-monophosphate preferential exoribonucleolysis. To achieve exoribonucleolytic processivity, the 5'-monophosphate-binding pocket and nucleotide +4 binding site anchor RNA within the catalytic track; the 5'-capping residue Leu37 of the sandwich pocket coupled with the 5'-monophosphate-binding pocket are dedicated to translocating and controlling the RNA orientation for each exoribonucleolytic cycle. The processive-exoribonucleolysis mechanism was verified as conserved in bacterial RNase J and also exposes striking parallels with the non-homologous eukaryotic 5'-3' exoribonuclease, Xrn1. The findings in this work shed light on not only the molecular mechanism of the RNase J family, but also the evolutionary convergence of divergent exoribonucleases.

Summary Mycobacterium tuberculosis (Mtb) has a proteasome system that is essential for its ability to cause lethal infections in mice. A key component of the system is the proteasomal adenosine triphosphatase (ATPase) Mpa, which captures, unfolds, and translocates protein substrates into the Mtb proteasome core particle for degradation. Here, we report the crystal structures of near full-length hexameric Mtb Mpa in apo and ADP-bound forms. Surprisingly, the structures revealed a ubiquitin-like β-grasp domain that precedes the proteasome-activating carboxyl terminus. This domain, which was only found in bacterial proteasomal ATPases, buries the carboxyl terminus of each protomer in the central channel of the hexamer and hinders the interaction of Mpa with the proteasome core protease. Thus, our work reveals the structure of a bacterial proteasomal ATPase in the hexameric form, and the structure finally explains why Mpa is unable to stimulate robust protein degradation in vitro in the absence of other, yet-to-be-identified co-factors.

Acrolein has been reported to have diverse toxic effects on various organs, including the reproductive system. However, little is known regarding the effects of maternal acrolein exposure on testicular steroidogenesis in male offspring. The present study investigated the effects of acrolein on fetal testosterone production and associated genes. Pregnant Sprague‑Dawley rats were intraperitoneally injected with vehicle (normal saline) or 1, 2 or 5 mg/kg acrolein from gestational day (GD) 14‑20, and fetal testes were examined on GD 21. Fetal body and testicular weights were markedly reduced in pups following exposure to high doses of acrolein (5 mg/kg) in late pregnancy. Notably, in utero exposure of 5 mg/kg acrolein significantly decreased the testicular testosterone level and downregulated the expression levels of steroidogenic acute regulatory protein (StAR) and 3β‑hydroxysteroid dehydrogenase (3β‑HSD), whereas the levels of other steroidogenic enzymes, including scavenger receptor class B, cholesterol side‑chain cleavage enzyme and steroid 17 alpha‑hydroxylase/17,20 lyase, were unaffected. Furthermore, the 3β‑HSD immunoreactive area in the interstitial region of the fetal testes was reduced at a 5 mg/kg dose, whereas the protein expression levels of 4‑hydroxynonenalwere dose‑dependently increased following maternal exposure to acrolein. mRNA expression levels of insulin‑like factor 3, a critical gene involved in testicular descent, were unaltered following maternal acrolein exposure. Taken together, the results of the present study suggested that maternal exposure to high doses of acrolein inhibited fetal testosterone synthesis, and abnormal expression of StAR and 3β‑HSD may be associated with impairment of the steroidogenic capacity.

Increasing researches have focused on the biological functions of long noncoding RNAs (lncRNAs) in human cancers. HOXA11-AS, a widely known lncRNA, has been confirmed to be involved in the progression of several cancers, including gastric cancer (GC). Whereas, the detailed mechanism of this lncRNA in GC remains to be further illuminated. The abundances of HOXA11-AS, miR-148a and WNT1 in GC tissues and cell lines were examined by qRT-PCR. Clinicopathological and Kaplan-Meier survival analyses were determined to explore the relationship between HOXA11-AS expression and outcomes of patients. Transwell assay was performed for the evaluation of cell migration and invasion. Bioinformatics, dual-luciferase reporter and RNA immunoprecipitation assays were employed to analyze the correlation between HOXA11-AS and miR-148a or miR-148a and WNT1. The protein levels of WNT1 and β-catenin were assessed by western blot assay. Results showed that HOXA11-AS and WNT1 expression levels were upregulated, while miR-148a level was downregulated in GC tissues and cell lines relative to matched controls. Elevated expression of HOXA11-AS was associated with increased tumor size, lymph node metastasis, advanced TNM stage, as well as reduced survival of GC patients. HOXA11-AS induced migration and invasion of GC cells through serving as a molecular sponge for miR-148a. Furthermore, miR-148a inactivated WNT1/β-catenin signaling pathway via directly targeting WNT1. HOXA11-AS increased WNT1/β-catenin pathway activity, which was abolished by miR-148a overexpression in GC cells. In conclusion, overexpression of HOXA11-AS contributed to migration and invasion of GC cells via activation of WNT1/β-catenin signaling pathway through repressing miR-148a, providing a prospective therapeutic target for GC.

A novel strain, YIM 131921T, was isolated from a Physcia sp. lichen collected from the South Bank Forest of the Baltic Sea. The strain is Gram-negative, catalase positive and oxidase negative, strictly aerobic, asporogenous, non-motile and reddish brown in colour. The temperature and pH for growth were found to be 20–30 °C (optimum 28 °C) and pH 6.5–12.0 (optimum pH 7.0 ± 0.5). No growth was observed in the presence of NaCl. Based on 16S rRNA gene sequence similarity, strain YIM 131921T shares high similarities with Rubellimicrobium roseum YIM 48858T (98.3%), followed by Rubellimicrobium mesophilum MSL-20T (96.8%), Rubellimicrobium aerolatum 5715S-9T (96.1%) and Rubellimicrobium thermophilum DSM 16684T (96.0%). Phylogenetic trees showed YIM 131921T forms a cluster with type strains of the genus Rubellimicrobium. The predominant cellular fatty acids (> 20%) were identified as summed feature 8 (C18:1ω7c) and C16:0. Q-10 was found to be the predominant respiratory ubiquinone. The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, glycolipid, phospholipids and an unidentified aminolipid. The DNA G + C content of the draft genome sequence is 66.6 mol%. Strain YIM 131921T showed an average nucleotide identity value of 80.3% and a digital DNA–DNA hybridizations value of 26.1% with the reference strain R. roseum YIM 48858T based on draft genome sequences. Based on comparative analyses of phenotypic, molecular, chemotaxonomic data and genomic comparisons, strain YIM 131921T is concluded to represent a novel species of the genus Rubellimicrobium, for which the name Rubellimicrobium rubrum sp. nov. is proposed. The type strain is YIM 131921T (= CGMCC 1.13958T = NBRC 114054T = KCTC 72461T).

Aminotransferases (ATs) are important biocatalysts for the synthesis of chiral amines because of their capability of introducing amino group into ketones or keto acids as well as their high enantioselectivity, high regioselectivity and no requirement of external addition of cofactor. Among all ATs, branched-chain amino acid aminotransferase (BCAT) can reversibly catalyse branched-chain amino acids (BCAAs), including L-valine, L-leucine, and L-isoleucine, with α-ketoglutaric acid to form the corresponding ketonic acids and L-glutamic acid. Alternatively, BCATs have been used for the biosynthesis of unnatural amino acids, such as L-tert-leucine. In the present study, the BCAT from Pseudomonas sp. (PsBCAT) was cloned and expressed in Escherichia coli for biochemical and structural analyses. The optimal reaction temperature and pH of PsBCAT were 40 °C and 8.5, respectively. PsBCAT exhibited a comparatively broader substrate spectrum, and showed remarkably high activity with L-leucine, L-valine, L-isoleucine and L-methionine with activities of 105 U/mg, 127 U/mg, 115 U/mg and 98 U/mg, respectively. Additionally, PsBCAT had activities with aromatic L-amino acids, L-histidine, L-lysine, and L-threonine. To analyse the catalytic mechanism of PsBCAT with the broad substrate spectrum, the crystal structure of PsBCAT was also determined. Finally, conjugated with the ornithine aminotransferase (OrnAT) from Bacillus subtilis, the coupled system was applied to the preparation of L-tert-leucine with 83% conversion, which provided an approximately 2.7-fold higher yield than the single BCAT reaction.IMPORTANCEDespite the enormous potential of BCATs, the vast majority of enzymes still lack suitably broad substrate scope and activity, thus new sources and novel enzymes are currently being investigated. Here, we described a previously uncharacterized PsBCAT, which showed a surprisingly wide substrate range and was more active towards BCAAs. This substrate promiscuity is unique for the BCAT family and could prove useful in industrial applications. Based on the determined crystal structure, we found some differences in the organization of the substrate binding cavity, which may influence the substrate specificity of the enzyme. Moreover, we demonstrated efficient biocatalytic asymmetric synthesis of L-tert-leucine using a coupling system, which can be used to remove the inhibitory by-product, and to shift the reaction equilibrium towards the product formation. In summary, the structural and functional characteristics of PsBCAT were analysed in detail, and this information will play an important role in the synthesis of chiral amino acids and will be conducive to industrial production of enantiopure chiral amines by aminotransferase.

Rv3899c is a hypothetical protein fromMycobacterium tuberculosiswhich is conserved across mycobacteria. It is predicted to be secreted and has been found in culture filtrates. It has been proposed as a potential vaccine candidate; however, its biological function is unknown. Here, the global structure of Rv3899c184–410, a fragment of Rv3899c, is reported. The structure resembles the shell of a sea snail, and its N- and C-termini form two relatively independent compact domains: an α/β/α sandwich folding domain and an α-helix bundle domain. There are no reported protein structures for any Rv3899c homologues; this structure provides the first structural glimpse of a new protein family consisting of Rv3899c and its homologues.

Psychrophilic enzymes play crucial roles in cold adaptation of microbes and provide useful models for studies of protein evolution, folding, and dynamic properties. We examined the crystal structure (2.2-Å resolution) of the psychrophilic β-glucosidase BglU, a member of the glycosyl hydrolase 1 (GH1) enzyme family found in the cold-adapted bacterium Micrococcus antarcticus . Structural comparison and sequence alignment between BglU and its mesophilic and thermophilic counterpart enzymes (BglB and GlyTn, respectively) revealed two notable features distinct to BglU: (i) a unique long-loop L3 (35 versus 7 amino acids in others) involved in substrate binding and (ii) a unique amino acid, His299 (Tyr in others), involved in the stabilization of an ordered water molecule chain. Shortening of loop L3 to 25 amino acids reduced low-temperature catalytic activity, substrate-binding ability, the optimal temperature, and the melting temperature ( T m ). Mutation of His299 to Tyr increased the optimal temperature, the T m , and the catalytic activity. Conversely, mutation of Tyr301 to His in BglB caused a reduction in catalytic activity, thermostability, and the optimal temperature (45 to 35°C). Loop L3 shortening and H299Y substitution jointly restored enzyme activity to the level of BglU, but at moderate temperatures. Our findings indicate that loop L3 controls the level of catalytic activity at low temperatures, residue His299 is responsible for thermolability (particularly heat lability of the active center), and long-loop L3 and His299 are jointly responsible for the psychrophilic properties. The described structural basis for the cold adaptedness of BglU will be helpful for structure-based engineering of new cold-adapted enzymes and for the production of mutants useful in a variety of industrial processes at different temperatures.

We aim to explore the associations of fascin-1 and cadherin-17 in gastric cancer (GC) to the clinicopathologic features and prognosis of GC. Case group included 204 GC tissues while control group comprised 204 paired adjacent cancer tissues. Expressions of fascin-1 and cadherin-17 were measured with immunohistochemistry and western blot and then analyzed statistically in relation to clinicopathologic features and survival time. Survival curve was drawn by Kaplan-Meier method, and independent prognostic factors were identified with Cox proportional hazards regression model. Fascin-1 was positively expressed in 45.1 % of GC tissues and in 27.5 % of adjacent cancer tissues, respectively (P

Background. In recent studies, aberrant expression of various microRNAs (miRNAs) is reported to be associated with gastric cancer metastasis.Method. Overexpression construct and inhibitor of hsa-miR-376c-3p were expressed in human gastric adenocarcinoma cell line SGC-7901. The expression level of tumor related genes was detected by qPCR, western blot, and immunostaining. Cell apoptosis was determined by flow cytometry. Xenograft of SGC-7901 cells was used to elucidate the function of hsa-miR-376c-3p in gastric tumor growthin vivo.Result. Expression of hsa-miR-376c-3p was detected in SGC-7901 cells. Downregulation of hsa-miR-376c-3p increased the expression level of BCL-2 and decreased the expression of smad4 and BAD. On the contrary, overexpression of hsa-miR-376c-3p increased the expression of BAD and smad4, while it led to the decreasing expression level of BCL-2. Overexpression of hsa-miR-376c-3p also promoted cell apoptosisin vitroand inhibited gastric tumor growthin vivo. Furthermore, the expression of BCL-2 was higher and expression of smad4 and BAD was lower in tumor tissue than the tissue adjacent to tumor from gastric cancer patients.Conclusion. This study demonstrated that hsa-miR-376c-3p plays an important role in the inhibition of gastric tumor growth and tumor related gene expression bothin vitroandin vivo.