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MicroRNAs (miRNAs) have emerged as key players in host–pathogen interaction and many virus-encoded miRNAs have been identified (computationally and/or experimentally) in a variety of organisms. A novel Bombyx mori nucleopolyhedrosis virus (BmNPV)-encoded miRNA miR-415 was previously identified through high-throughput sequencing. In this study, a BmNPV-miR-415 expression vector was constructed and transfected into BmN cells. The differentially expressed protein target of rapamycin isoform 2 (TOR2) was observed through two-dimensional gel electrophoresis and mass spectrometry. Results showed that TOR2 is not directly a target gene of BmNPV-miR-415, but its expression is up-regulated by BmNPV-miR-415 via Bmo-miR-5738, which could be induced by BmNPV. [ABSTRACT FROM AUTHOR]

[Display omitted] • OfPGRP8 exhibits the highest transcript level in fat body. • The mRNA level of OfPGRP8 significantly increases upon invading microbes. • OfPGRP8 exhibits intensive binding ability to peptidoglycans. • OfPGRP8 inhibits the growth of Staphylococcus aureus and Micrococcus luteus. • Knockdown of OfPGPR8 leads to a marked decrease in the expression of AMPs genes. Peptidoglycan recognition proteins (PGRPs) discriminate and bind peptidoglycans by acting as pattern recognition receptors (PRRs) in insects, and function pivotal roles in innate immune response. In the present study, we cloned a full-length PGRP gene designed as OfPGRP8 from the Asian corn borer, Ostrinia furnacalis (Guenée). Its mRNA exhibited the highest abundance in fat body, and its expression level upregulated dramatically after bacterial challenges. Purified recombinant OfPGRP8 exhibited intensive binding capacity to peptidoglycans from Staphylococcus aureus and Micrococcus luteus. Additionally, recombinant OfPGRP8 could inhibit the growth of S. aureus , M. luteus. However, recombinant OfPGRP8 could not cause agglutination of S. aureus , M. luteus or Escherichia coli. Furthermore, we also demonstrated that OfPGRP8 may be involved in modulating the signaling pathway of antimicrobial peptides (AMPs) synthesis. In sum, our results provided evidence that OfPGRP8 discriminates peptidoglycans from microbes and acts as a PRR to initiate downstream immune signaling pathways. [ABSTRACT FROM AUTHOR]

The silkworm, Bombyx mori , is an important model organism of lepidopteran insects, and its testis is a main male reproductive organ and spermatogenesis place. Studying the testis helps to understand the mechanisms of genetic sterility of lepidopteran insects and to achieve sterile insect technique (SIT) for pest control. Herein, we performed a comparative transcriptome analysis of testes between three biological replicates of the GMS mutant and wild strain 898WB, respectively. In total, 1872 up-regulated genes and 1823 down-regulated genes were identified in the testis of the GMS mutant. Several genes contribute significantly to spermatogenesis and testis development, such as "serine/threonine protein kinase", "organic cation transporter protein", "tyrosine protein kinase", "lncRNAs" and "immune-associated genes". The KEGG pathway analysis shows that the DEGs were annotated to 123 pathways, and 10 pathways were significantly enriched, such as "metabolic pathway", "biosynthesis of amino acids", and "phagosome–lysosome pathway", which are associated with testis development and spermatogenesis. The results of the qPCR expression were consistent with the RNA-seq data, which shows that the RNA-seq results were accurate. The DEGs of the testes between GMS mutant and 898WB were screened by RNA-Seq technology, which provides a reliable reference to understand the molecule mechanism of male sterility of the GMS mutant. Based on testis transcriptome sequencing, we compared the gene expression patterns between the testis of the GMS mutant and 898WB. Functional analyses of DEGs demonstrate that the infertility of the GMS mutant may be attributed to distinct gene expression during spermatogenesis. Our study provides basic data for understanding the mechanism of reproductive development and developing insect sterility technique. [Display omitted] • This is the first report about transcriptome studies on male sterile line (GMS), Bombyx mori. • Several genes related to spermatogenesis were abnormally expressed in the GMS mutant. • KEGG pathways which associated with spermatogenesis were significantly enriched. [ABSTRACT FROM AUTHOR]

The acoustic spin pumping (ASP) stands for the generation of a spin voltage from sound waves in a ferromagnet/paramagnet junction. In this letter, we propose and demonstrate a method for pure detection of the ASP, which enables the separation of sound-wave-driven spin currents from the spin Seebeck effect due to the heating of a sample caused by a sound-wave injection. Our demonstration using a Pt/YIG/PZT sample shows that the ASP signal in this structure measured by a conventional method is considerably offset by the heating signal and that the pure ASP signal is one order of magnitude greater than that reported in the previous study. [ABSTRACT FROM AUTHOR]

• We explored multiple coexisting factors associated with NTDs. • High tHcy coexisting with inadequate free T4 was 3 times more at risk of NTDs. • High Hcy might affect metabolism of thyroid hormones through modifying genes. This study tested the hypothesis that abnormal maternal metabolism of both homocysteine and thyroid hormone network in pregnant women is associated with neural tube defects (NTDs) in a part of China with high NTD prevalence. A case–control study was performed between 2007 and 2009 in Lüliang Mountains, Shanxi Province. This study included 83 pregnant women who had fetuses with NTDs (cases) and 90 pregnant women with normal fetuses (controls). In addition, a cell model to illustrate the epidemiological findings was established. Fetuses of mother who had both high total homocysteine (tHcy) and inadequate free thyroxine were 3 times more at risk of developing NTDs (adjusted odds ratio = 3.5; 95 % confidence interval = 1.2–10.4; cases vs. controls) using multivariate logistic regression models. Furthermore, biological interaction between metabolisms of Hcy and thyroid hormones was demonstrated in vitro. In homocysteine thiolactone of a metabolite of Hcy-treated mouse embryonic neural stem NE4C cells, genes (Bmp7, Ctnnb1, Notch 1, Gli2, and Rxra) related to both neural tube closure and thyroid hormone network were shown to be regulated by H3K79 homocysteinylation, which increased their expression levels. The effect of maternal serum high tHcy on risk of developing NTDs is depended on maternal serum level of thyroxine. Meanwhile, a higher level of tHcy might also affect both maternal metabolism of thyroid hormone and neural tube closure in embryogenesis through homocysteinylation of histones. [ABSTRACT FROM AUTHOR]

Abstract: Bisphenol A (BPA) is a representatively endocrine disruptor, which shows a highly toxic effect on life system. Its potential toxicity on plants that are the primary producers in earth’s ecosystem is not well documented. Here, the effects of BPA on the growth, photosynthesis, content of chlorophyll (Chl), initial fluorescence (F 0), maximal photochemical efficiency (F v/F m), effective quantum yield of photosystem II (Φ PSII) and photosynthetic electron transport rate (ETR) in soybean seedlings were investigated by using gas exchange measurement system and chlorophyll fluorometer to understand the toxic effect of BPA on plants. It was found that when soybean seedlings were treated with BPA at the low concentration (1.5mgL−1), the growth indices (the plant height, fresh and dry weights of stems, fresh and dry weights of leaves, leaf area) were increased obviously compared with those of the control, which was not related with the photosynthesis, the content of Chl and the chlorophyll fluorescence. When soybean seedlings were treated with BPA at the high concentrations (7.0, 12.0, 17.2 and 50.0mgL−1), the growth indices, net photosynthetic rate, the content of chlorophyll, F v/F m, Φ PSII, ETR were decreased, while F 0 was increased, compared with those of the control. Obviously, BPA at the high concentrations showed the toxic effect on soybean seedlings. The results from correlation analysis indicated that the inhibition in the growth of soybean seedlings treated with BPA was related to the decrease in photosynthesis because of the decrease in the content of chlorophyll and the change in chlorophyll fluorescence. [Copyright &y& Elsevier]

Abstract: In this work, indium zinc oxide (IZO) films have been deposited on a polyethylene terephthalate substrate coated with an SiO x film. Based on a comparative investigation of an IZO monolayer and an IZO/SiO x multilayer, it is shown that oxygen has a great effect on the electrical properties of the thin films. A mechanism is described to explain the influence of the introduced SiO x buffer layer. It is considered that an interfacial layer has come into being at the interface between the SiO x layer and IZO layer, and the properties of this layer have been evaluated. Moreover, the electrical properties of the IZO/SiO x multilayer have been successfully improved by controlling the oxygen content of the interfacial layer. [Copyright &y& Elsevier]

The electronic structure and magnetic properties of Co/Al2O3/Co with O-terminated and Al-terminated interface models of different thicknesses are investigated by first-principles discrete variational method with the local-spin-density approximation. Our calculations results show that the magnetic moment of interface Co layer is enhanced for Al-terminated and weakened for O-terminated interface compared with that of bulk Co. For O-terminated interface models, spin polarization at Fermi energy of Co layer at interface exhibits negative and becomes positive for O layer at interface. In contrast, both Co and Al layers at interface possess negative SP for the Al-terminated interface models. We have also found that TMR ratio of Al-terminated interface models is much larger than that of O-terminated interface. In addition, the change of SP with the thickness of insulating layer is in a similar way as that of magnetic moment. [Copyright &y& Elsevier]

Flexoelectric effect in ferroelectric epitaxy films has attracted significant attention due to its theoretical and application potentials. However, the origin and control of flexoelectric field is not yet fully understood. In this study, we investigated the surface structure of single-domain epitaxial BiFeO 3 films with a (111) orientation using grazing incidence X-ray diffraction and X-ray reflection methods. We observed large strain gradient on the surface of the BiFeO 3 films, which produces a giant flexoelectric field up to 82.55 MV/m at the surface of 15 nm thick BiFeO 3 film. Additionally, we discovered the presence of a surface layer with lower electron density than that of the underlying BiFeO 3 layer, which is related to the flexoelectric field. Moreover, with increased film thickness, we found the strain distribution along growth direction of the film is changed, thereby leads to the decreased flexoelectric field and increasing thickness of the surface layer respectively. These findings convinced the coupling between the strain state and surface structure, changing the thickness of the ferroelectric film and consequently the strain state is an efficient way to adjust the flexoelectric field and the surface structure. [ABSTRACT FROM AUTHOR]

The natural colorful cuticles of insects play important roles in many physiological processes. Pigmentation is a physiological process with a complex regulatory network whose regulatory mechanism remains unclear. Bombyx mori pigmentation mutants are ideal materials for research on pigmentation mechanisms. The purple quail-like (q-l p ) and brown quail-like (q-l b ) mutants originated from plain silkworm breeds 932VR and 0223JH respectively exhibit similar cuticle pigmentation to that of the quail mutant. The q-l p mutant also presents a developmental abnormality. In this study, genes controlling q-l p and q-l b mutants were located on chromosome 8 by positional cloning. Then the neuropeptide gene orcokinin (OK) was identified to be the major gene responsible for two quail-like mutants. The B. mori orcokinin gene (BommoOK) produces two transcripts, BommoOKA and BommoOKB, by alternative splicing. The CRISPR/Cas9 system and orcokinin peptides injection were used for further functional verification. We show a novel function of BommoOKA in inhibiting pigmentation, and one mature peptide of orcokinin A, OKA_type2, is the key factor in pigmentation inhibition. These results provide a reference for studying the function of orcokinin and are of theoretical importance for studying the regulatory mechanism of pigmentation. The purple quail-like (q-l p ) and brown quail-like (q-l b ) mutants originated from plain silkworm breeds 932VR and 0223JH respectively. By positional cloning, the neuropeptide gene orcokinin (OK) was identified to be the major gene responsible for two quail-like mutants, one base deletion in q-l p mutant and a 4.3 kb fragment insertion in q-l b mutant. The quail-like pigmentation appeared after BommoOK gene knockout by CRISPR/cas9, and on the contrast the mutant characteristics was rescued by injecting peptides of BommoOK, which completely indicated that orcokinin gene was the key factor of two quail-like mutants. Image 1 • We confirmed that BommoOK involves in the regulation of pigmentation by its mature peptide OKA_type2. • A new method, peptide injection was established for exploration of peptide function. • The results are of theoretical importance for the study of mechanism of insect pigmentation and function of BommoOK gene. [ABSTRACT FROM AUTHOR]

Aldose reductase (AR) is a rate-limiting enzyme in the polyol pathway and is also the key enzyme involved in diabetic complications. The silkworm purple quail-like mutant ( q-l p ) exhibits pigmented dots on its epidermis. The q-l p mutant also shows developmental abnormalities and decreased vitality. In this study, fat bodies from the q-l p mutant and the wildtype 932VR strain were subjected to two-dimensional gel electrophoresis (2-DE) analysis, and the Bombyx mori AR ( BmAR ) protein was found to be significantly downregulated in the q-l p mutant. The expression of BmAR at the mRNA level was also significantly downregulated, as verified through quantitative reverse transcription PCR (qRT-PCR). Knockdown of the expression of BmAR via RNAi resulted in a reduction of silkworm weight. The sorbitol level in q-l p was significantly lower than in the wildtype. These results suggested that the BmAR gene is closely related to the development of the q-l p mutant. Investigation of the cause of BmAR downregulation in the q-l p mutant could contribute to revealing the function of AR in insects and offers a new method of identifying AR inhibitors for the treatment of diabetic complications. [ABSTRACT FROM AUTHOR]

Congenital hydrocephalus is heterogeneous in its etiology, and in addition to a genetic component, has been shown to be caused by environmental factors. Until now, however, no methylation alterations of target genes have been connected with congenital hydrocephalus in humans. Frizzled 3(FZD3) is a planar cell polarity (PCP) gene required for PCP signaling. Partial restoration of frizzled 3 activities in FZD3 mutant mice results in hydrocephalus. To analyze the possible roles of epigenetic modifications of the FZD3 gene in congenital hydrocephalus pathogenesis, DNA methylation in the promoter region of FZD3 was assayed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Gene expression and chromatin accessibility were also determined to assess the role of methylation alterations. Our study found methylation levels of the FZD3 gene were increased in congenital hydrocephalus, especially in males (10.57±3.90 vs. 7.08±0.94, p=0.001). Hypermethylation of FZD3 increased congenital hydrocephalus risk, with an odds ratio of 10.125 (p=0.003). Aberrant methylation modification of FZD3 altered both chromatin structure in this region and FZD3 expression levels. Totally, aberrant methylation modification of the FZD3 gene increases the risk of congenital hydrocephalus by altering chromatin structure and disturbing gene expression. [ABSTRACT FROM AUTHOR]

• The amino acid sequences of DpPGRP-S1 resembled canonical characteristic features of PGRPs. • The transcript level of DpPGRP-S1 was differentially expressed in tissues, abundantly in fat body. • Recombinant PGRP-S1 demonstrated distinct binding affinity to different microorganism cell wall components. • Recognition and binding of PGRP-S1 to microbes might be related to proPO stimulation cascade. Recognition of invading foreign exogenous pathogen is the first step to initiate the innate immune response of insects, which accomplished by the pattern recognition receptors (PRRs). Peptidoglycan recognition proteins (PGRPs) serve as an important type of PRRs, which activate immune response by detecting peptidoglycan of microbial cell wall. In this study, we have cloned the full-length cDNA of PGRP gene called PGRP-S1 from the Diaphania pyloalis (Walker). The open reading frame (ORF) of D. pyloalis PGRP-S1 encodes 211 amino acids which containing a secretion signal peptide and a canonical PGRP domain. Multisequence alignment revealed that PGRP-S1 possess the amino acid residues responsible for zinc binding and amidase activity. D. pyloalis PGRP-S1 exhibited the highest transcript level in fat body and followed in head. The mRNA concentration dramatically increased after an injection of Escherichia coli or Micrococcus luteus. Purified recombinant PGRP-S1 exhibit binding ability to peptidoglycans from Staphylococcus aureus or Bacillus subtilis and cause intensive agglutination of E. coli , M. luteus or S. aureus in the presence of zinc ions. Furthermore, phenoloxidase activity significantly increased when the plasma from larvae was incubated with recombinant PGPR-S1 and peptidoglycans from B. subtilis or M. luteus simultaneously. These results implied that PGRP-S1 was a member involving the prophenoloxidase activation pathway. Overall, our results indicated that D. pyloalis PGRP-S1 serve as a PRR to participate in the recognition of foreign pathogen and prophenoloxidase pathway stimulation. [ABSTRACT FROM AUTHOR]

Abstract: The egg stage is an important stage in the silkworm (Bombyx mori) life cycle. Normal silkworm eggs are usually short, elliptical, and laterally flattened, with a sometimes hollowed surface on the lateral side. However, the eggs laid by homozygous recessive “Ming” lethal egg mutants (l-em ) lose water and become concaved around 1h, ultimately exhibiting a triangular shape on the egg surfaces. We performed positional cloning, and narrowed down the region containing the gene responsible for the l-em mutant to 360kb on chromosome 10 using 2287 F2 individuals. Using expression analysis and RNA interference, the best l-em candidate gene was shown to be BmEP80. The results of the inverse polymerase chain reaction showed that an ~1.9kb region from the 3′ untranslated region of BmVMP23 to the forepart of BmEP80 was replaced by a >100kb DNA fragment in the l-em mutant. Several eggs laid by the normal moths injected with BmEP80 small interfering RNAs were evidently depressed and exhibited a triangular shape on the surface. The phenotype exhibited was consistent with the eggs laid by the l-em mutant. Moreover, two-dimensional gel electrophoresis showed that the BmEP80 protein was expressed in the ovary from the 9th day of the pupa stage to eclosion in the wild-type silkworm, but was absent in the l-em mutant. These results indicate that BmEP80 is responsible for the l-em mutation. [Copyright &y& Elsevier]

Abstract: Elevated Programmed Death-1 (PD-1) expression can inhibit T cell activity and is a potential barrier to achieving persisting and optimal immunity via therapeutic vaccination. Using a direct lymph node-targeted vaccination procedure that enabled uncoupling of synthetic peptide (signal 1, TCR-mediated) and adjuvant (signal 2, non-TCR-mediated), we evaluated the impact of varied doses of Toll-like receptor (TLR)-9 ligand CpG oligodeoxynucleotide (ODN) adjuvant on epitope-specific CD8+ T cell-associated PD-1 expression. Peptide vaccination without adjuvant yielded CD8+ T cells with significantly elevated PD-1 expression. This conferred impaired function ex vivo, but was reversible by antibody-mediated PD-1 blockade. By comparison, peptide vaccination with escalating doses of CpG ODN adjuvant yielded higher magnitudes of CD8+ T cells with progressively lower PD-1 expression and greater ex vivo function. CpG ODN adjuvant in context of titrated peptide doses for vaccination yielded the lowest overall PD-1 expression levels, demonstrating that fine-tuning both TCR-independent (adjuvant dose) and -dependent (antigen dose) stimuli can synergize to co-regulate PD-1 expression on epitope-specific CD8+ T cells. These data hint at strategies to elicit PD-1low CD8+ T cells using TLR-9 ligand adjuvants, and also shed light on the PD-1-regulated homeostasis of CD8+ T cells. [Copyright &y& Elsevier]

Abstract: DNA vaccines or peptides are capable of inducing specific immunity; however, their translation to the clinic has generally been problematic, primarily due to the reduced magnitude of immune response and poor pharmacokinetics. Herein, we demonstrate that a novel immunization strategy, encompassing sequential exposure of the lymph node milieu to plasmid and peptide in a heterologous prime-boost fashion, results in considerable MHC class I-restricted immunity in mice. Plasmid-primed antigen expression was essential for the generation of a population of central memory T cells, expressing CD62L and low in PD-1, with substantial capability to expand and differentiate to peripheral memory and effector cells, following subsequent exposure to peptide. These vaccine-induced T cells dominated the T cell repertoire, were able to produce large amounts of chemokines and pro-inflammatory cytokines, and recognized tumor cells effectively. In addition to outlining a feasible and effective method to transform plasmid DNA vaccination into a potentially viable immunotherapeutic approach for cancer, this study sheds light on the mechanism of heterologous prime-boost and the considerable heterogeneity of MHC class I-restricted T cell responses. [Copyright &y& Elsevier]