Dioscin induces osteosarcoma cell apoptosis by upregulating ROS-mediated P38 MAPK signaling.

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. Many patients with osteosarcoma readily develop resistance to chemotherapy and have an extremely dismal prognosis. Dioscin, a saponin, is known to exhibit potent anticancer activities and induce cellular death of a variety of cancer types. However, the inhibitory effect of dioscin on osteosarcoma cells and its underlying mechanisms have not been fully elucidated. We investigated the responses of human U2-OS and MG63 osteosarcoma cells to dioscin with regard to proliferation, apoptosis, migration, and invasion, and studied the effect of dioscin on MAPK-related proteins by western blot analysis assays. Dioscin inhibited osteosarcoma cell proliferation, migration, and invasion. Moreover, it induced osteosarcoma cell apoptosis via reactive oxygen species (ROS)-dependent apoptotic signaling. N-acetylcysteine, a reactive oxygen species inhibitor, suppressed dioscin-induced apoptosis, indicating that ROS play an essential role in dioscin-induced apoptosis. Western blot analysis assays showed that p38 MAPK was upregulated after dioscin treatment, and that dioscin induced apoptosis by upregulating ROS-mediated p38 MAPK signaling. Our study suggests that dioscin possesses antitumor activities against human osteosarcoma cells, inhibits osteosarcoma cell proliferation, migration and invasion, and induces osteosarcoma cell apoptosis through upregulating ROS-mediated p38 MAPK signaling. This study may provide a new therapeutic strategy and potential clinical applications for the treatment of osteosarcoma.

Unique bacterial communities associated with components of an artificial aquarium ecosystem and their possible contributions to nutrient cycling in this microecosystem.

In order to better understand the bacterial distribution characteristics in a whole microecosystem, the bacterial communities in different components of an artificial aquarium (i.e., plants, fishes, sand and water) were characterized using high throughput sequencing of bacterial 16S rRNA genes. Across all samples, 2873 operational taxonomic units were identified and assigned to 771 genera in 36 phyla. In a principle coordinate analysis, samples clustered according to their origin, indicating that bacterial communities from the same component were most similar. Further taxonomic analysis revealed that most dominant genera, even those with the similar functions, were biased to one component: Nitrospira and Rhodobacter were mainly abundant in plant samples; Rhodococcus, Serratia, Ralstonia, Sphingobacterium and Pseudomonas were most common in sand samples; Cetobacterium and Aeromonas dominated fish samples; and Flavobacterium, Alpinimonas and Limnobacter were especially common in water samples. Functional predictions performed by PICRUSt and the dominant genera exhibited that bacteria detected in each component could participate in all nutrient cycles in the aquarium. However, those involved in carbon and nitrogen cycling were most common in plant and fish samples, while phosphate metabolism-related pathways were more abundant in sand and water samples. Moreover, the aquarium plants, in association with their bacterial communities might be the most important component in the aquarium, as indicated by their highest bacterial richness and diversity. This study adds to our understanding on the differences in the microbiome of different components and their possible contributions to nutrient cycling in a self-sustaining aquarium.

Galectin-3 deficiency protects lipopolysaccharide-induced chondrocytes injury via regulation of TLR4 and PPAR-γ-mediated NF-κB signaling pathway.

The aim of the present study was to identify the functional role of galectin-3 (Gal-3) in lipopolysaccharide (LPS)-induced injury in ATDC5 cells and to explore the probable molecular mechanisms. Here, we identified that LPS is sufficient to enhance the expression of Gal-3 in ATDC5 cells. In addition, repression of Gal-3 obviously impeded LPS-stimulated inflammation damage as exemplified by a reduction in the release of inflammatory mediators interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α, as well as the production of nitric oxide and prostaglandin E2 (PGE2) concomitant with the downregulation of matrix metalloproteinases (MMP)-13 and MMP-3 expression in ATDC5 cells after LPS administration. Moreover, ablation of Gal-3 dramatically augmented cell ability and attenuated cell apoptosis accompanied by an increase in the expression of antiapoptotic protein Bcl-2 and a decrease in the expression of proapoptotic protein Bax and caspase-3 in ATDC5 cells subjected with LPS. Importantly, we observed that forced expression of TLR4 or blocked PPAR-γ with the antagonist GW9662 effectively abolished Gal-3 inhibition-mediated anti-inflammatory and antiapoptosis effects triggered by LPS. Mechanistically, depletion of Gal-3 prevents the NF-κB signaling pathway. Taken together, these findings indicated that the absence of Gal-3 exerted chondroprotective properties dependent on TLR4 and PPAR-γ-mediated NF-κB signaling, indicating that Gal-3 functions as a protector in the development and progression of osteoarthritis.

Sphingobacterium athyrii sp. nov., a cellulose- and xylan-degrading bacterium isolated from a decaying fern (Athyrium wallichianum Ching).

Assessment of the bacterial diversity associated with a decaying fern, Athyrium wallichianum Ching, revealed the presence of a novel bacterial strain named M46T. It was Gram-stain-negative, rod-shaped, non-motile and aerobic with cellulose and xylan degradation abilities. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain M46T was affiliated to the genus Sphingobacterium, exhibiting the highest sequence similarity of 97.9 % to Sphingobacterium ginsenosidimutans THG 07T, Sphingobacterium canadense CR11T and Sphingobacterium detergens6.2 ST. Multilocus sequence analysis (MLSA) based on concatenated sequences of the rpoB, cpn60 and 16S rRNA genes showed that strain M46T clustered together with S. canadense CR11T. The genome of strain M46T had a G+C content of 40.6 mol% and chromosome of 6 853 865 bp. Average nucleotide identity (ANI) between strain M46T and S. detergens 6.2 ST and S. siyangense SY1T was 85.1 and 78.1 %, respectively. DNA-DNA relatedness values among strain M46T and other closely related Sphingobacterium species were <70 %. ANI and DNA-DNA relatedness findings strongly supported M46T as a putative novel strain of Sphingobacterium. The predominant fatty acids of strain M46T were iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and iso-C17 : 0 3-OH, and MK-7 was the dominant isoprenoid quinone. The polar lipid profile of strain M46T contained phosphatidylethanolamine as the dominant component, while minor amounts of phosphoglycolipid, one unidentified aminophospholipid, two unidentified phospholipids and four unidentified lipids were also detected. Based on 16S rRNA gene sequence similarities, MLSA results, genomic characteristics, and phenotypic and biochemotaxonomic analyses, strain M46T is considered to represent a novel species in the genus Sphingobacterium, for which the name Sphingobacterium athyrii sp. nov. is proposed. The type strain is M46T (=CGMCC 1.13466T=JCM 32543T).

Identification of co-expression modules and pathways correlated with osteosarcoma and its metastasis.

BACKGROUND: Osteosarcoma is the most common bone tumor that occurs in children. METHODS: To identify co-expression modules and pathways correlated with osteosarcoma and its clinical characteristics, we performed weighted gene co-expression network analysis (WGCNA) on RNA-seq data of osteosarcoma with 52 samples. Then we performed pathway enrichment analysis on genes from significant modules. RESULTS: A total of 5471 genes were included in WGCNA, and 16 modules were identified. Module-trait analysis identified that a module involved in microtubule bundle formation, drug metabolism-cytochrome P450, and IL-17 signaling pathway was negatively correlated with osteosarcoma and positively correlated with metastasis; a module involved in DNA replication was positively correlated with osteosarcoma; a module involved in cell junction was positively correlated with metastasis; and a module involved in heparin binding negatively correlated with osteosarcoma. Moreover, expression levels in four of the top ten differentially expressed genes were validated in another independent dataset. CONCLUSIONS: Our analysis might provide insight for molecular mechanisms of osteosarcoma.

Stenosing tenosynovitis : Evaluation of percutaneous release with a specially designed needle vs. open surgery.

PURPOSE: The purpose of this study was to evaluate the effectiveness of conventional open surgery and percutaneous release with a specially designed needle for treating stenosing tenosynovitis in terms of cure, relapse and complication rates. METHODS: In this study 89 fingers from 76 patients were randomly assigned and allocated to one of the treatment groups. A total of 37 patients were treated with open surgery in group 1 and 39 patients with percutaneous release using a specially designed needle in group 2. A patient-based 4-inch visual analogue scale (VAS), Quinnell grading (QG), disability of arm shoulder and hand (DASH) score and finger total joint range of motion (FTROM) score were evaluated before treatment and after 7, 30 and 180 days. When finger QG scores were equal or greater than 2 points at follow-up at 180 days this was defined as recurrence.. RESULTS: There were no significant differences between the two groups (P > 0.05) in terms of VAS, DASH and QG scores and the degree of FTROM. At 7 days all the data were significantly different (p < 0.05) compared with preoperative data, 30 days was significantly different (p < 0.05) compared with 7 days while at 180 days no significant differences could be found (p > 0.05) compared with 30 days. The recurrence rate in group 1 was 4.65% and 6.55% in group 2. CONCLUSION: The percutaneous release and open surgery methods displayed similar effectiveness regarding the cure and recurrence of trigger finger disorder. The use of a specially designed needle for release is a safe and reliable method.

Connexin 43 Modulates Osteogenic Differentiation of Bone Marrow Stromal Cells Through GSK-3beta/Beta-Catenin Signaling Pathways.

BACKGROUND/AIMS: Bone marrow stromal cells (BMSCs) are multipotent precursors that give rise to osteoblasts, and contribute directly to bone formation. Connexin 43 (Cx43) is the most ubiquitous gap junction protein expressed in bone cell types, and plays crucial roles in regulating intercellular signal transmission for bone development, differentiation and pathology. However, the precise role and mechanism of Cx43 in BMSCs are less known. Here, we investigate the function of Cx43 in osteogenic differentiation of BMSCs in vitro. METHODS: BMSCs were isolated by whole bone marrow adherent culture. Knock down of Cx43 was performed by using lentiviral transduction of Cx43 shRNA. BMSCs were induced to differentiate by culturing in a-MEM, 10% FBS, 50 µM ascorbic acid, 10 mM beta-glycerophosphate, and 100 nM dexamethasone. Alkaline phosphatase (ALP) activity and alizarin red S staining were used to evaluate osteogenic differentiation in calcium nodules. Target mRNAs and proteins were analyzed by using real-time quantitative PCR (qPCR) and western blotting. RESULTS: Cx43 expression markedly increased during osteogenic differentiation. Osteogenic differentiation was suppressed following lentiviral-mediated knockdown of Cx43 expression, as judged by decreased levels of Runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), osteocalcin (Bglap), Osterix (Osx), alkaline phosphatase (ALP) activity and the number of calcium nodules in response to osteogenic differentiation stimuli. Knock down of Cx43 reduced the level of phosphorylation of GSK-3beta at Ser9 (p-GSK-3beta), resulting in decreased beta-catenin expression and activation. Furthermore, treatment of Cx43-knockdown cells with lithium chloride (LiCl), a GSK-3beta inhibitor, reduced osteogenic differentiation and decreased GSK-3beta levels, as well as partially rescued levels of both total and activated beta-catenin. CONCLUSION: These findings indicate that Cx43 positively modulates osteogenic differentiation of BMSCs by up-regulating GSK-3beta/beta-catenin signaling pathways, suggesting a potential role for Cx43 in determining bone mass and bone mineral density by modulating osteogenesis.

Ruthenium-Catalyzed Decarboxylative C-H Alkenylation in Aqueous Media: Synthesis of Tetrahydropyridoindoles.

We disclose herein a Ru(II)-catalyzed decarboxylative and oxidative coupling of N-substituted indolyl carboxylic acids with broad substrate scope in an aqueous solution. This method provides a sustainable and efficient access to synthesize various indole-fused cyclohexanyl acetic acids under mild conditions.

Identification of thienopyridine carboxamides as selective binders of HIV-1 trans Activation Response (TAR) and Rev Response Element (RRE) RNAs.

Small organic molecules that can selectively bind to RNA with specificity are relatively rare. Here we report the synthesis, biochemical and structural studies of thienopyridine carboxamide derivatives with the capacity of selectively recognizing and binding with HIV-1 TAR and RRE RNAs that are essential elements for viral replication.

Moss habitats distinctly affect their associated bacterial community structures as revealed by the high-throughput sequencing method.

To better understand the factors that influence the distribution of bacteria associated with mosses, the communities inhabiting in five moss species from two different habitats in Beijing Songshan National Nature Reserve were investigated using the high-throughput sequencing method. The sequencing was performed based on the bacterial 16S rRNA and 16S rDNA libraries. Results showed that there are abundant bacteria inhabiting in all the mosses sampled. The taxonomic analysis of these bacteria showed that they mainly consisted of those in the phyla Proteobacteria and Actinobacteria, and seldom were from phylum Armatimonadetes, Bacteroidetes and Firmicutes. The hierarchical cluster tree, based on the OTU level, divided the bacteria associated with all samples into two branches according to the habitat types of the host (terrestrial and aquatic). The PCoA diagram further divided the bacterial compositions into four groups according to both types of habitats and the data sources (DNA and RNA). There were larger differences in the bacterial community composition in the mosses collected from aquatic habitat than those of terrestrial one, whether at the DNA or RNA level. Thus, this survey supposed that the habitat where the host was growing was a relevant factor influencing bacterial community composition. In addition, the bacterial community detected at the RNA level was more sensitive to the habitat of the growing host, which could also be proved by the significantly differences in the predicted function by PICRUSt and the metabolically active dominant genera between different groups. This study expands the knowledge about the interactions between mosses and microbes.

AgI /TMG-Promoted Cascade Reaction of Propargyl Alcohols, Carbon Dioxide, and 2-Aminoethanols to 2-Oxazolidinones.

Chemical valorization of CO2 to access various value-added compounds has been a long-term and challenging objective from the viewpoint of sustainable chemistry. Herein, a one-pot three-component reaction of terminal propargyl alcohols, CO2 , and 2-aminoethanols was developed for the synthesis of 2-oxazolidinones and an equal amount of α-hydroxyl ketones promoted by Ag2 O/TMG (1,1,3,3-tetramethylguanidine) with a TON (turnover number) of up to 1260. By addition of terminal propargyl alcohol, the thermodynamic disadvantage of the conventional 2-aminoethanol/CO2 coupling was ameliorated. Mechanistic investigations including control experiments, DFT calculation, kinetic and NMR studies suggest that the reaction proceeds through a cascade pathway and TMG could activate propargyl alcohol and 2-aminoethanol through the formation of hydrogen bonds and also activate CO2 .

Illumina sequencing of bacterial 16S rDNA and 16S rRNA reveals seasonal and species-specific variation in bacterial communities in four moss species.

In order to better understand the factors that influence bacterial diversity and community composition in moss-associated bacteria, a study of bacterial communities in four moss species collected in three seasons was carried out via high-throughput sequencing of 16S rDNA and 16S rRNA. Moss species included Cratoneuron filicinum, Pylaisiella polyantha, Campyliadelphus polygamum, and Grimmia pilifera, with samples collected in May, July, and October 2015 from rocks at Beijing Songshan National Nature Reserve. In total, the bacterial richness and diversity were high regardless of moss species, sampling season, or data source (DNA vs. RNA). Bacterial sequences were assigned to a total of 558 OTUs and 279 genera in 16 phyla. Proteobacteria and Actinobacteria were the two most abundant phyla, and Cellvibrio, Lapillicoccus, Jatrophihabitans, Friedmanniella, Oligoflexus, and Bosea the most common genera in the samples. A clustering algorithm and principal coordinate analysis revealed that C. filicinum and C. polygamum had similar bacterial communities, as did P. polyantha and G. pilifera. Metabolically active bacteria showed the same pattern in addition to seasonal variation: bacterial communities were most similar in summer and autumn, looking at each moss species separately. In contrast, DNA profiles lacked obvious seasonal dynamics. A partial least squares discriminant analysis identified three groups of samples that correlated with differences in moss species resources. Although bacterial community composition did vary with the sampling season and data source, these were not the most important factors influencing bacterial communities. Previous reports exhibited that mosses have been widely used in biomonitoring of air pollution by enriching some substances or elements in the moss-tag technique and the abundant moss associated bacteria might also be important components involved in the related biological processes. Thus, this survey not only enhanced our understanding of the factors which influence microbial communities in mosses but also would be helpful for better use and development of the moss-tag technique in the environmental biomonitoring.

Illumina sequencing-based community analysis of bacteria associated with different bryophytes collected from Tibet, China.

BACKGROUND: Previous studies on the bacteria associated with the bryophytes showed that there were abundant bacteria inhabited in/on these hosts. However, the type of bacteria and whether these discriminate between different bryophytes based on a particular factor remains largely unknown. RESULTS: This study was designed to analyze the biodiversity and community of the bacteria associated with ten liverworts and ten mosses using Illumina-sequencing techniques based on bacterial 16S rRNA gene. A total of 125,762 high quality sequences and 437 OTUs were obtained from twenty bryophytes. Generally, there were no obvious differences between the richness of bacteria associated with liverworts and mosses; however, the diversity was significantly higher in liverworts than in mosses. The taxonomic analyses showed that there were abundant bacteria inhabited with each bryophyte and those primarily detected in all samples were within the phyla Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Armatimonadetes and Planctomycetes. In addition, bacteria assigned to Chloroflexi, Fibrobacteres, Gemmatimonadetes, Chlamydiae, group of TM6 and WCHB1-60 also appeared in part of the bryophytes. The assigned bacteria included those adapted to aquatic, anaerobic and even extreme drought environments, which is consistent with the bryophyte transition from aquatic to terrestrial conditions. Of them, approximately 10 recognized genera were shared by all the samples in a higher proportion, such as Burkholderia, Novosphingobium, Mucilaginibacter, Sorangium, Frankia, Frondihatitans, Haliangium, Rhizobacter, Granulicella and Hafnia, and 11 unclassified genera were also detected in all samples, which exhibited that large amounts of unclassified bacteria could interact with the bryophytes. The Heatmap and Principle Coordinate Analyses showed that bacteria associated with six mosses displayed a higher community similarity. Notably, the bacteria associated with another four mosses exhibited higher similarity with the ten liverworts. CONCLUSIONS: The result of further analysis of the bacterial community in different bryophytes revealed that the phylogeny of hosts might portray a strong influence on the associated bacterial community and that niche also played important roles when the hosts were phylogenetically more similar. Further studies are needed to confirm the role of phylogeny on bacterial communities and determine the level of influence on predicting which bacteria is associated with the host.

Paenibacillus marchantiophytorum sp. nov., isolated from the liverwort Herbertus sendtneri.

A Gram-stain-positive, aerobic, motile, rod-shaped, endospore-forming bacterium, designated strain R55T, was isolated from the liverwort Herbertus sendtneri growing at Gawalong glacier, Tibet, and characterized by using a polyphasic taxonomic approach. The major fatty acids of strain R55T were anteiso-C15 : 0, C16 : 0 and iso-C16 : 0. Diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were the predominant polar lipids and occurred along with two unidentified aminophospholipids, one unidentified phospholipid and one unidentified aminolipid. Strain R55T contained MK-7 as the dominant menaquinone and meso-diaminopimelic acid as the major diagnostic diamino acid in the cell-wall peptidoglycan. The G+C content of the genomic DNA was 36.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain R55T was affiliated to species of the genus Paenibacillus, and was related most closely to Paenibacillus ferrarius CY1T (97.1 % similarity). However, the DNA-DNA relatedness between this strain and strain R55T was only 44.1 %. Based on phenotypic, chemotaxonomic and phylogenetic properties, strain R55T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus marchantiophytorum sp. nov. is proposed. The type strain is R55T ( = CGMCC 1.15043T = DSM 29850T).

The elevated preoperative neutrophil-to-lymphocyte ratio predicts poor prognosis in intrahepatic cholangiocarcinoma patients undergoing hepatectomy.

A high preoperative peripheral blood neutrophil-to-lymphocyte ratio (NLR) has been reported to be a predictor of poor survival in patients with various cancers. The aim of this study was to evaluate the predictive significance of the NLR in patients undergoing hepatectomy for intrahepatic cholangiocarcinoma (ICC). From 2005 to 2011, 322 patients who underwent hepatectomy for ICC were enrolled in this retrospective study. Clinicopathological parameters, including NLR, were evaluated to identify predictors of overall and recurrence-free survival after hepatectomy. The best cutoff for NLR was 2.49, and 177 of 322 patients (54.9 %) had an NLR ≥ 2.49. The 5-year survival rate after hepatectomy was 51.1 % in patients with NLR < 2.49 and 24.8 % in those with NLR ≥ 2.49 (P = 0.0001). Univariate analyses revealed that NLR was significantly associated with recurrence-free survival (RFS) and overall survival (OS; both P < 0.05). Multivariable analyses revealed that elevated NLR independently predicted poorer OS (P = 0.003, hazard ratio [HR] = 1.600). In summary, our results indicate that elevated NLR is a promising independent predictor of poor survival after hepatectomy in patients with ICC.

Chryseobacterium takakiae sp. nov., a member of the phylum Bacteroidetes isolated from Takakia lepidozioides.

A Gram-stain-negative, rod-shaped and non-endospore-forming bacterium, designated strain AG1-2(T), was isolated from Takakia lepidozioides collected from the Gawalong glacier in Tibet, China and characterized using a polyphasic taxonomic approach. The predominant fatty acids of strain AG1-2(T) were iso-C15 : 0 (36.0 %), iso-C17:0 3-OH (20.2 %), summed feature 9 (iso-C17:1ω9c and/or C16:0 10-methyl, 16.4%) and summed feature 3 (C16:1ω7c and/or C16:1ω6c, 11.1%). The major polar lipids were phosphatidylethanolamine, three unidentified aminolipids and two unidentified lipids. Strain AG1-2(T) contained MK-6 as the dominant menaquinone, and the genomic DNA G+C content was 37.3 mol%. The phylogenetic analysis based on the 16S rRNA gene sequences showed that strain AG1-2(T) was affiliated to species of the genus Chryseobacterium, and its closest related species were Chryseobacterium taiwanense Soil-3-27(T), Chryseobacterium hispalense AG13(T), Chryseobacterium camelliae THG C4-1(T) and Chryseobacterium taeanense PHA3-4(T) with a sequence similarity of 98.0, 97.8, 97.3 and 97.1%, respectively. However, the DNA-DNA relatedness values between these strains and strain AG1-2(T) were 29, 21, 21 and 45%, respectively. Based on phylogenetic inference and phenotypic data, strain AG1-2(T) is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium takakiae sp. nov. is proposed. The type strain is AG1-2(T) ( = CGMCC 1.12488(T) = DSM 26898(T)).

Chryseobacterium polytrichastri sp. nov., isolated from a moss (Polytrichastrum formosum), and emended description of the genus Chryseobacterium.

A Gram-stain negative, rod-shaped and non-endospore forming bacterium, designated strain YG4-6(T), was isolated from Polytrichastrum formosum collected from Gawalong glacier in Tibet, China and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain YG4-6(T) were identified as iso-C15:0 (29.3 %), summed feature 3 (C16:1 ω7c and/or C16:1 ω6c as defined by MIDI, 23.5 %) and iso-C17:0 3-OH (16.5 %). The major polar lipids were found to consist of five unidentified aminolipids and three unidentified lipids. Strain YG4-6(T) was found to contain MK-6 as the dominant menaquinone and the G+C content of its genomic DNA was determined to be 37.3 mol%. The phylogenetic analysis based on 16S rRNA gene sequences showed that strain YG4-6(T) is affiliated to Chryseobacterium species, and its closest related species were Chryseobacterium aahli T68(T) (97.9 % sequence similarity), Chryseobacterium zeae JM-1085(T) (97.8 % sequence similarity), Chryseobacterium yeoncheonense DCY67(T) (97.6 % sequence similarity) and Chryseobacterium soldanellicola NBRC 100864(T) (97.2 % sequence similarity). However, the DNA-DNA relatedness values between these strains and strain YG4-6(T) were found to be clearly below 70 %. Based on the phylogenetic inference and phenotypic data, strain YG4-6(T) is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium polytrichastri sp. nov. is proposed. The type strain is YG4-6(T) (=CGMCC 1.12491(T) = DSM 26899(T)). An emended description of the genus Chryseobacterium is also proposed.

Paenibacillus herberti sp. nov., an endophyte isolated from Herbertus sendtneri.

Strain R33(T), an endophyte recovered from Herbertus sendtneri, was identified as representing a novel species of the genus Paenibacillus by using a polyphasic taxonomic approach. The novel strain was observed to be a Gram-stain positive, aerobic, rod-shaped, motile and endospore-forming bacterium. The major polar lipids of strain R33(T) were identified as diphosphatidylglycerol, phosphatidylethanolamine, along with lesser amounts of phosphatidylglycerol, three unidentified aminophospholipids, two unidentified phospholipids and two unidentified lipids. The predominant isoprenoid quinone was identified as MK-7. The major fatty acids (>8.0 %) were found to be anteiso-C15:0 (40.0 %), C16:1 ω11c (9.4 %), C16:1 ω7c alcohol (8.5 %) and C16:0 (8.2 %). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The G+C content of genomic DNA was determined to be 56.9 mol%. The 16S rRNA gene sequence similarities of strain R33(T) to other Paenibacillus species ranged from 91.6 to 97.2 %, with high similarities to Paenibacillus humicus PC-147(T) and Paenibacillus pasadenensis SAFN-007(T). The phylogenetic analyses based on 16S rRNA gene sequences and the partial rpoB gene confirmed that strain R33(T) belongs to the genus Paenibacillus. However, strain R33(T) shows differential molecular characteristics compared to other related Paenibacillus species based on 16S rDNA-RFLP analyses; the DNA-DNA relatedness values between strain R33(T) and P. humicus PC-147(T), and that between strain R33(T) and P. pasadenensis SAFN-007(T), were 35.0 ± 2.0 and 41.4 ± 0.9 %, respectively. Based on its phenotypic, chemotaxonomic and phylogenetic properties, strain R33(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus herberti is proposed (type strain R33(T) = CGMCC 1.15042(T) = DSM 29849(T)).

Cnuella takakiae gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from Takakia lepidozioides.

A Gram-staining-negative, rod-shaped and non-spore-forming bacterium, designated strain RG1-1(T), was isolated from Takakia lepidozioides collected from Gawalong glacier in Tibet, China, and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain RG1-1(T) were iso-C(15 : 0) (19.8%), summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c, 17.0%), C(16 : 0 (9.9)%) and iso-C(17 : 0) 3-OH (9.4%); its major polar lipids were phosphatidylethanolamine, four unidentified aminolipids, one unidentified phospholipid, one unidentified aminoglycolipid, one unidentified glycolipid, and three unidentified lipids. Strain RG1-1(T) contained MK-7 as the dominant menaquinone, and the G+C content of its genomic DNA was 49.1 mol%. Strain RG1-1(T) exhibited the highest 16S rRNA gene sequence similarity (91.8%) with Flavisolibacter ginsengiterrae Gsoil 492(T) and Flavisolibacter ginsengisoli Gsoil 643(T). Phylogenetic analysis showed that strain RG1-1(T) was a member of the family Chitinophagaceae, phylum Bacteroidetes. On the basis of 16S rRNA gene sequence analysis, and phenotypic and chemotaxonomic data, strain RG1-1(T) is considered to represent a novel species of a novel genus, for which the name Cnuella takakiae gen. nov., sp. nov. is proposed. The type strain is RG1-1(T) ( = CGMCC 1.12492(T) = DSM 26897(T)).

Mucilaginibacter polytrichastri sp. nov., isolated from a moss (Polytrichastrum formosum), and emended description of the genus Mucilaginibacter.

A Gram-staining-negative, rod-shaped, non-spore-forming bacterium, designated strain RG4-7(T), was isolated from the moss Polytrichastrum formosum collected from Gawalong glacier in Tibet, China, and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain RG4-7(T) were summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid, two unidentified aminolipids and one unidentified lipid. Strain RG4-7(T) contained MK-7 as the dominant menaquinone and the G+C content of its genomic DNA was 39.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain RG4-7(T) was affiliated to species of the genus Mucilaginibacter, and its closest relative was Mucilaginibacter jinjuensis YC7004(T) (97.0 % sequence similarity). However, the DNA-DNA relatedness between this strain and strain RG4-7(T) was only 49.1±3.7 %. Based on phylogenetic inference and phenotypic data, strain RG4-7(T) is considered to represent a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter polytrichastri sp. nov. is proposed. The type strain is RG4-7(T) ( = CGMCC 1.12493(T) = DSM 26907(T)). An emended description of the genus Mucilaginibacter is also proposed.