<Emphasis Type="Italic">Brevibacterium anseongense</Emphasis> sp. nov., isolated from soil of ginseng field

Gram-positive, aerobic, non-motile, pale-yellow, and rodshaped bacterium, designated as Gsoil 188^T, was isolated from the soil of a ginseng field in Pocheon, South Korea. A phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that the strain formed a distinct lineage within the genus Brevibacterium and was most closely related to B. epidermidis NBRC 14811^T (98.4%), B. sediminis FXJ8.269^T (98.2%), B. avium NCFB 3055^T (98.1%), and B. oceani BBH7^T (98.1%), while it shared less than 98.1% identity with the other species of this genus. The DNA G + C content was 68.1 mol%. The predominant quinone was MK-8(H₂). The major fatty acids were anteiso-C_15:0 and anteiso-C_17:0. The cell wall peptidoglycan of strain Gsoil 188^T contained meso-diaminopimelic acid. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and an unidentified aminolipid. The physiological and biochemical characteristics, low DNA-DNA relatedness values, and taxonomic analysis allowed the differentiation of strain Gsoil 188^T from the other recognized species of the genus Brevibacterium. Therefore, strain Gsoil 188^T represents a novel species of the genus Brevibacterium, for which the name Brevibacterium anseongense sp. nov. is proposed, with the type strain Gsoil 188^T (= KACC 19439^T = LMG 30331^T).     

Genome-wide analysis and stress-responsive expression of CCCH zinc finger family genes in <Emphasis Type="Italic">Brassica rapa</Emphasis>

Background

Ubiquitous CCCH nucleic acid-binding motif is found in a wide-variety of organisms. CCCH genes are involved in plant developmental processes and biotic and abiotic stress responses. Brassica rapa is a vital economic crop and classical model plant of polyploidy evolution, but the functions of CCCH genes in B. rapa are unclear.

Results

In this study, 103 CCCH genes in B. rapa were identified. A comparative analysis of the chromosomal position, gene structure, domain organization and duplication event between B. rapa and Arabidopsis thaliana were performed. Results showed that CCCH genes could be divided into 18 subfamilies, and segmental duplication might mainly contribute to this family expansion. C-X_7/8-C-X₅-C₃-H was the most commonly found motif, but some novel CCCH motifs were also found, along with some loses of typical CCCH motifs widespread in other plant species. The multifarious gene structures and domain organizations implicated functional diversity of CCCH genes in B. rapa. Evidence also suggested functional redundancy in at least one subfamily due to high conservation between members. Finally, the expression profiles of subfamily-IX genes indicated that they are likely involved in various stress responses.

Conclusion

This study provides the first genome-wide characterization of the CCCH genes in B. rapa. The results suggest that B. rapa CCCH genes are likely functionally divergent, but mostly involved in plant development and stress response. These results are expected to facilitate future functional characterization of this potential RNA-binding protein family in Brassica crops.

     

Effect of substrate stiffness on proliferation and differentiation of periodontal ligament stem cells

Objectives

The aim of this study was to understand the effect of substrate stiffness (a mechanical factor of the extracellular matrix) on periodontal ligament stem cells (PDLSCs) and its underlying mechanism.

Materials and methods

Elastic substrates were fabricated by mixing 2 components, a base and curing agent in proportions of 10:1, 20:1, 30:1 or 40:1. PDLSC morphology was observed using scanning electron microscopy (SEM). Cell proliferation and differentiation were assessed after PDLSCs was cultured on various elastic substrates. Data were analysed using one‐way ANOVA.

Results

SEM revealed variations in the morphology of PDLSCs cultured on elastic substrates. PDLSC proliferation increased with substrate stiffness (P < .05). Osteogenic differentiation of PDLSCs was higher on stiff substrates. Notch pathway markers were up‐regulated in PDLSCs cultured on stiff substrates.

Conclusions

Results suggested that the osteogenic differentiation of PDLSCs might be promoted by culturing them in a stiffness‐dependent manner, which regulates the Notch pathway. This might provide a new method of enhancing osteogenesis in PDLSCs.

     

Synthesis of oxidative metabolites of CRA13 and their analogs: Identification of CRA13 active metabolites and analogs thereof with selective CB2R affinity

CRA13; a peripheral dual CB₁R/CB₂R agonist with clinically proven analgesic properties, infiltrates into CNS producing adverse effects due to central CB₁R agonism. Such adverse effects might be circumvented by less lipophilic compounds with attenuated CB₁R affinity. Metabolism produces less lipophilic metabolites that might be active metabolites. Some CRA13 oxidative metabolites and their analogues were synthesized as less lipophilic CRA13 analogues. Probing their CB₁R and CB₂R activity revealed the alcohol metabolite 8c as a more potent and more effective CB₂R ligand with attenuated CB₁R affinity relative to CRA13. Also, the alcohol analogue 8b and methyl ester 12a possessed enhanced CB₂R affinity and reduced CB₁R affinity. The CB₂R binding affinity of alcohol analogue 8b was similar to CRA13 while that of methyl ester 12a was more potent. In silico study provided insights into the possible molecular interactions that might explain the difference in the elicited biological activity of these compounds.     

Cells scaffold complex for Intervertebral disc Anulus Fibrosus tissue engineering: in vitro culture and product analysis

The study was designed to investigate feasibility of tissue culture in vitro utilizing static culture method. Annulus fibrosus cells obtained from spine of rabbits were cultured. Results showed that fibrous tissue infiltration could be detected in shallow layer. With extended time, tissue infiltration depth increased, but there were still a large amount of holes in central part. Fibrous tissue infiltration was detected in the control side products and inner infiltration wasn't obvious. Hydroxyproline content of the control side products gradually increased with extended culture time. Hydroxyproline content of the control side products in the third and fourth month was significantly higher than that in the first month, but lower than those of the experimental side products and normal annulus fibrosus cells. DNA content of the control side products in the third and fourth month was significantly increased compared to the first month. DNA content of the control side products at each phase point was significantly lower than that of the experimental side and normal annulus fibrosus cells. Furthermore, there was lower expression levels of the type I, II collagen mRNA and protein in the experimental side scaffolds compared to the control side product. This study demonstrates the successful formation of Intervertebral disc Anulus Fibrosus in vitro by static culture method.     

NGF 与Neuritin 在脑外伤伴四肢骨折病人血清中的表达及意义

目的:通过检测脑外伤患者、脑外伤合并骨折患者、骨折患者及正常人外周血中NGF,neuritin不同时间段的表达,根据其含量的变化,判断与骨折愈合速度的相关性,寻找骨折愈合的关键因子。方法:收集单纯脑外伤、单纯骨折患者各80例、脑外伤合并骨折患者60例、健康体检人群20例。选取外伤后3天、10天、2周抽取所有实验对象的静脉血,应用ELISA技术测定标本中NGF与Neuritin的含量。结果:损伤后每个时间段里,患者血清中NGF、neuritin含量有不同程度升高,均高于正常对照组,其中又以脑外伤合并骨折组最高。血清NGF在骨折合并脑外伤组伤后第3天含量明显升高,为(0.86±0.21),伤后10天为(1.47±0.29),14天为(2.0 7±0.21),脑外伤合并四肢骨折组neuritin血清含量在伤后第3天略有升高为(83.47±18.85),10天(108.50±31.65),2周(91.86±21.12).脑外伤合并骨折病人血清NGF、neuritin表达明显高于其他对照组,差别均有统计学意义(P<0.05)。结论:脑外伤合并骨折病人血清NGF、neuritin表达明显高于其他对照组,说明与骨折愈合有着密切的相关性,两种因子可能在骨折愈合修复过程中共同起作用,从而,推测可能是脑外伤后骨折愈合加速的重要因素。     

The amino‐terminal tail of Hxt11 confers membrane stability to the Hxt2 sugar transporter and improves xylose fermentation in the presence of acetic acid

Hxt2 is a glucose repressed, high affinity glucose transporter of the yeast Saccharomyces cerevisiae and is subjected to high glucose induced degradation. Hxt11 is a sugar transporter that is stably expressed at the membrane irrespective the sugar concentration. To transfer this property to Hxt2, the N‐terminal tail of Hxt2 was replaced by the corresponding region of Hxt11 yielding a chimeric Hxt11/2 transporter. This resulted in the stable expression of Hxt2 at the membrane and improved the growth on 8% d ‐glucose and 4% d ‐xylose. Mutation of N361 of Hxt11/2 into threonine reversed the specificity for d ‐xylose over d ‐glucose with high d ‐xylose transport rates. This mutant supported efficient sugar fermentation of both d ‐glucose and d ‐xylose at industrially relevant sugar concentrations even in the presence of the inhibitor acetic acid which is normally present in lignocellulosic hydrolysates. Biotechnol. Bioeng. 2017;114: 1937–1945. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.     

Cathepsin B inhibitory activities of three new phthalate derivatives isolated from seahorse, Hippocampus Kuda Bleeler

Three new phthalate acid derivatives, 2,12-diethyl-11-methylhexadecyl 2-ethyl-11-methylhexadecyl phthalate (1), 2-ethyldecyl 2-ethylundecyl phthalate (2), and bis(2-ethyldodecyl) phthalate (3), were isolated from seahorse, Hippocampus Kuda Bleeler, together with a known natural analog bis(2-ethylheptyl) phthalate (4). The structures of these compounds were elucidated mainly by means of the comprehensive analysis of their NMR spectroscopic data. The four phthalate derivatives showed dose-dependent cathepsin B inhibitions activities with IC(50) values of 0.13 mM (1), 0.21 mM (2), 0.18 mM (3), and 0.29 mM (4), respectively.