Fluorescence studies on the interaction of a synthetic signal peptide and its analog with liposomes

AbstractThe N-terminal signal sequence of glucitol permease of Escherichia coli (Gut22: MIETITHGAEWFIGLFQKGGEC) and its analog (Gut22Ana: MIETITPGAVWFIGLFQKGGEC) were synthesized. The analog had a Pro residue substituted for the His at the 7th position of Gut22 and a Val residue substituted for the Glu at the 10th position. Previous studies indicated that due to its structural rigidity, the interaction of Gut22Ana with lipid bilayer was much weaker than that of Gut22 (Wang, Q.D., Cui, D.F. and Lin, Q.S. (1996) Science in China (Series C) 39, 395–405). To further probe the location of the tryptophan residues of the peptides in lipid bilayer, the membrane penetration depth of the tryptophan residue of Gut22 was measured using spin-labeled phospholipids, and fluorescence quenching of the peptides by iodide and acrylamide in the presence and absence of phosphatidylserine/phosphatidylcholine liposomes were also studied. Fluorescent labeling of the peptides enabled the study of their association with membrane by fluorospectrophotometry. In the presence of liposomes, the peptides were protected from reaction with chymotrypsin, indicating that the peptide incorporated into the membrane. However, dithionite, which acts external to the membrane, reacted with the peptide, showing that the peptides did not translocate across lipid bilayer spontaneously

Experimental Study on the Mechanical Behaviors of PVA-ECC after Freeze-Thaw Cycles

yesIn order to study the mechanical behaviors of engineered cementitious composites (ECC) reinforced with various types of polyvinyl alcohol (PVA) fibers and different fiber volume fractions after the freeze-thaw cycles, the rapid freeze-thaw method was used to test the mass loss ratios, longitudinal relative dynamic elastic modulus, compressive strength and flexural strength. The results showed that specimens incurred more serious damage with the increasing of freeze-thaw cycles; however their performance would be improved by fiber type and dosage. Mass loss rate of JPA (specimen with 2% volume content of JP fiber) was lower than JPB (specimen with 1% volume content of JP fiber). Furthermore, the mass loss rate of JPB was lower than that of CPB (specimen with 1% volume content of CP fiber). The longitudinal relative dynamic elastic modulus of JPA was higher than that of JPB while the longitudinal relative dynamic elastic modulus of JPB was higher than that of CPB. In addition, the compressive strength and flexural strength decreased with the increasing of freeze-thaw cycles. Mechanical behaviors of specimens with fiber exhibited better strength than specimens without fiber. Based on the SL 211-2006 code for the design of hydraulic structures against ice and freezing action, JPA and JPB specimens are adequate for use in severe cold regions, while specimen CPA and CPB can be used in cold regions, specimen JPC only can be used in warm regions

Overexpression of BMI-1 Promotes Cell Growth and Resistance to Cisplatin Treatment in Osteosarcoma

Background: BMI-1 is a member of the polycomb group of genes (PcGs), and it has been implicated in the development and progression of several malignancies, but its role in osteosarcoma remains to be elucidated. Methodology/Principal Findings: In the present study, we found that BMI-1 was overexpressed in different types of osteosarcomas. Downregulation of BMI-1 by lentivirus mediated RNA interference (RNAi) significantly impaired cell viability and colony formation in vitro and tumorigenesis in vivo of osteosarcoma cells. BMI-1 knockdown sensitized cells to cisplatininduced apoptosis through inhibition of PI3K/AKT pathway. Moreover, BMI-1-depletion-induced phenotype could be rescued by forced expression of BMI-1 wobble mutant which is resistant to inhibition by the small interfering RNA (siRNA). Conclusions/Significance: These findings suggest a crucial role for BMI-1 in osteosarcoma pathogenesis

Comprehensive investigation and regulatory function of lncRNAs engaged in western honey bee larval immune response to Ascosphaera apis invasion

Ascosphaera apis is a fungal pathogen that exclusively infects bee larvae, causing chalkbrood disease, which results in severe damage for beekeeping industry. Long non-coding RNAs (lncRNAs) are versatile regulators in various biological processes such as immune defense and host-pathogen interaction. However, expression pattern and regulatory role of lncRNAs involved in immune response of bee host to A. apis invasion is still very limited. Here, the gut tissues of Apis mellifera ligustica 4-, 5-, and 6-day-old larvae inoculated by A. apis spores (AmT1, AmT2, and AmT3 groups) and corresponding un-inoculated larval guts (AmCK1, AmCK2, and AmCK3 groups) were prepared and subjected to deep sequencing, followed by identification of lncRNAs, analysis of differentially expressed lncRNAs (DElncRNAs), and investigation of competing endogenous RNA (ceRNA) network. In total, 3,746 A. m. ligustica lncRNAs were identified, including 78 sense lncRNAs, 891 antisense lncRNAs, 1,893 intergenic lncRNAs, 346 bidirectional lncRNAs, and 210 intronic lncRNAs. In the 4-, 5-, and 6- comparison groups, 357, 236, and 505 DElncRNAs were discovered. Additionally, 217, 129, and 272 DElncRNAs were respectively predicted to regulate neighboring genes via cis-acting manner, and these targets were associated with a series of GO terms and KEGG pathways of great importance, such as response to stimulus and Jak-STAT signaling pathway. Moreover, 197, 95, and 356 DElncRNAs were observed to target 10, eight, and 21 DEmiRNAs and further target 147, 79, and 315 DEmRNAs, forming complex regulatory networks. Further investigation suggested that these targets were engaged in several key cellular and humoral immune pathways, such as phagosome and MAPK signaling pathway. Ultimately, the expression trends of nine randomly selected DElncRNAs were verified by RT-qPCR, confirming the authenticity and reliability of our transcriptome data. Findings in this current work not only provide candidate DElncRNAs for functional study, but also lay a foundation for unclosing the mechanism underlying DElncRNA-regulated larval immune responses to A. apis invasion

First identification of long non-coding RNAs in fungal parasite Nosema ceranae

International audienceAbstractNosema ceranae is a unicellular fungal parasite of honey bees and causes huge losses for apiculture. Until present, no study on N. ceranae long non-coding RNAs (lncRNAs) was documented. Here, we sequenced purified spores of N. ceranae using strand-specific library construction and high-throughput RNA sequencing technologies. In total, 83 novel lncRNAs were predicted from N. ceranae spore samples, including lncRNAs, long intergenic non-coding RNAs (lincRNAs), and sense lncRNAs. Moreover, these lncRNAs share similar characteristics with those identified in mammals and plants, such as shorter length and fewer exon number and transcript isoforms than protein-coding genes. Finally, the expression of 12 lncRNAs was confirmed with RT-PCR, confirming their true existence. To our knowledge, this is the first evidence of lncRNAs produced by a microsporidia species, offering novel insights into basic biology such as regulation of gene expression of this widespread taxonomic group

Population genomic analysis reveals that homoploid hybrid speciation can be a lengthy process

This work was supported by grants from National key research and development program (2017YFC0505203), National Natural Science Foundation of China (grant numbers 31590821, 31670665, 91731301), National Key Project for Basic Research (2014CB954100), CAS “Light of West China” Program and Graduate Student’s Research and Innovation Fund of Sichuan University (2018YJSY007).An increasing number of species are thought to have originated by homoploid hybrid speciation (HHS), but in only a handful of cases are details of the process known. A previous study indicated that Picea purpurea, a conifer in the Qinghai–Tibet Plateau (QTP), originated through HHS from P. likiangensis and P. wilsonii. To investigate this origin in more detail, we analysed transcriptome data for 114 individuals collected from 34 populations of the three Picea species from their core distributions in the QTP. Phylogenetic, principal component and admixture analyses of nuclear SNPs showed the species to be delimited genetically and that P. purpurea was admixed with approximately 60% of its ancestry derived from P. wilsonii and 40% from P. likiangensis. Coalescent simulations revealed the best‐fitting model of origin involved formation of an intermediate hybrid lineage between P. likiangensis and P. wilsonii approximately 6 million years ago (mya), which backcrossed to P. wilsonii to form P. purpurea approximately one mya. The intermediate hybrid lineage no longer exists and is referred to as a “ghost” lineage. Our study emphasizes the power of population genomic analysis combined with coalescent analysis for reconstructing the stages involved in the origin of a homoploid hybrid species over an extended period. In contrast to other studies, we show that these stages can in some instances span a relatively long period of evolutionary time.PostprintPeer reviewe

Reticulate evolution within a spruce (Picea) species complex revealed by population genomic analysis

This work was supported by grants from National key research and development program (2017YFC0505203), National Natural Science Foundation of China (grant numbers 31590821, 31670665, 91731301), National Key Project for Basic Research (2014CB954100), “1000 Youth Talents Plan” of Yunnan Province and CAS “Light of West China” Program.The role of reticulation in the rapid diversification of organisms is attracting greater attention in evolutionary biology. Evidence of genetic exchange between diverging taxa is reported frequently, although most studies fail to show how hybridization and introgression contribute to the adaptation and differentiation of introgressed taxa. Here, we report a population genomics approach to test the role of hybridization and introgression in the evolution of the Picea likiangensis species complex, which comprises four taxa occurring in the biodiversity hotspot of the Hengduan-Himalayan mountains. Based on 84,793 SNPs detected in transcriptomes of 82 trees collected from 35 localities, we identified 18 hybrids (including backcrosses) distributed within the range boundaries of the four taxa. Coalescent simulations, for each pair of taxa and for all taxa taken together, rejected several tree-like divergence models and supported instead a reticulate evolution model with secondary contacts occurring during Pleistocene glacial cycles after initial divergence in the late Pliocene. Significant gene flow occurred among some taxa after secondary contact according to an analysis based on modified ABBA-BABA statistics that accommodated a rapid diversification scenario. A novel finding was that introgression between certain taxa can contribute to increasing divergence (and possibly reproductive isolation) between those taxa and other taxa within a complex at some loci. These results illuminate the reticulate nature of evolution within the P. likiangensis complex and highlight the value of population genomic data in detecting the effects of introgression in the rapid diversification of related taxa.PostprintPeer reviewe

Effects of excluding grazing on the vegetation and soils of degraded sparse-elm grassland in the Horqin Sandy Land, China

Livestock grazing is a crucial cause of vegetation degradation and desertification in sandy lands. The sparse-elm grassland of Horqin Sandy Land, China has suffered severe degradation of biodiversity and ecosystem services. Management to exclude grazing is often necessary for ecological restoration, especially in arid and semi-arid regions. We report effects on vegetation and soils in a 10-year experiment to exclude livestock, completely or seasonally, in comparison with a continuously grazed area in Horqin. Complete exclusion of grazing and restriction of grazing to summer both led to significantly increased plant cover and density relative to the grazed control. Species richness increased, reflected in higher Shannon-Wiener indices; only complete exclusion increased the Simpson diversity index, whereas Pielou evenness was significantly lowest under seasonal grazing. Exclosure treatments were also associated with improved soil texture, and increased water retention, available nitrogen, total nitrogen, total carbon and total phosphorus. Soil pH and C/N ratio were highest under the seasonal grazing regime. The results indicated that exclosure management indeed improved biodiversity and ecosystem services in an erosion-prone region. Although total exclosure was most effective in restoration of degraded sparse-elm grassland, seasonal grazing management was highly beneficial and represented a good compromise with resource utilization and economic development

Recent Advances in Synthetic Bioelastomers

This article reviews the degradability of chemically synthesized bioelastomers, mainly designed for soft tissue repair. These bioelastomers involve biodegradable polyurethanes, polyphosphazenes, linear and crosslinked poly(ether/ester)s, poly(ε-caprolactone) copolymers, poly(1,3-trimethylene carbonate) and their copolymers, poly(polyol sebacate)s, poly(diol-citrates) and poly(ester amide)s. The in vitro and in vivo degradation mechanisms and impact factors influencing degradation behaviors are discussed. In addition, the molecular designs, synthesis methods, structure properties, mechanical properties, biocompatibility and potential applications of these bioelastomers were also presented