Antimicrobial Peptide LL37 and MAVS Signaling Drive Interferon-ß Production by Epidermal Keratinocytes during Skin Injury.

Type 1 interferons (IFNs) promote inflammation in the skin but the mechanisms responsible for inducing these cytokines are not well understood. We found that IFN-ß was abundantly produced by epidermal keratinocytes (KCs) in psoriasis and during wound repair. KC IFN-ß production depended on stimulation of mitochondrial antiviral-signaling protein (MAVS) by the antimicrobial peptide LL37 and double stranded-RNA released from necrotic cells. MAVS activated downstream TBK1 (TANK-Binding Kinase 1)-AKT (AKT serine/threonine kinase 1)-IRF3 (interferon regulatory factor 3) signaling cascade leading to IFN-ß production and then promoted maturation of dendritic cells. In mice, the production of epidermal IFN-ß by LL37 required MAVS, and human wounded and/or psoriatic skin showed activation of MAVS-associated IRF3 and induction of MAVS and IFN-ß gene signatures. These findings show that KCs are an important source of IFN-ß and MAVS is critical to this function, and demonstrates how the epidermis triggers unwanted skin inflammation under disease conditions.

Dual layer chessboard metasurface sandwiched by a spin-on-carbon for spectral modulation.

Metasurfaces, composed by metals and dielectrics in periodical order with subwavelength pitches, are of great importance for their unique ability to abruptly manipulate optical fields. So far, all the reported metasurfaces are constructed by thermally deposited metals and dielectric films, based on semiconductor processes which are expensive and time-consuming. Inspired by the outstanding dry etch property of spin-on-carbon (SOC) as the interlayer material in CMOS technology, this paper proposes to utilize the SOC as the dielectric layer in a chessboard metasurface with dual layer of gold to form an array of local surface plasmonic resonators (localized surface plasmon resonance). Finite difference and time domain (FDTD) method is used to investigate the spectral characteristics in reflectance of the metasurface in both visible and short wavelengths of infrared light. Electron beam lithography is applied to generate the nanoscale chessboard pattern on ZEP520A, followed by a conventional oxygen-based plasma etch to form high aspect ratio nanopillar arrays in SOC with the feature width under 50 nm, and ended by a thermal deposition of gold to form self-aligned dual layer local surface plasmonic resonators (LSPRs). The measured reflectance spectra agree with the simulated. A wealth of optical properties, such as coupling induced modulations of spectra by LSPRs, are revealed and analyzed. These special modes result in tunable structural colors and wavelength-selective antireflection ability. To the best of our knowledge, this is the first time that SOC is applied in the construction of metasurfaces, which has great potential for next generation nanophotonic devices.

Regulation of integrin and extracellular matrix genes by HNRNPL is necessary for epidermal renewal.

Stratified epithelia such as the epidermis require coordinated regulation of stem and progenitor cell proliferation, survival, and differentiation to maintain homeostasis. Integrin-mediated anchorage of the basal layer stem cells of the epidermis to the underlying dermis through extracellular matrix (ECM) proteins is crucial for this process. It is currently unknown how the expression of these integrins and ECM genes are regulated. Here, we show that the RNA-binding protein (RBP) heterogeneous nuclear ribonucleoprotein L (HNRNPL) binds to these genes on chromatin to promote their expression. HNRNPL recruits RNA polymerase II (Pol II) to integrin/ECM genes and is required for stabilizing Pol II transcription through those genes. In the absence of HNRNPL, the basal layer of the epidermis where the stem cells reside prematurely differentiates and detaches from the underlying dermis due to diminished integrin/ECM expression. Our results demonstrate a critical role for RBPs on chromatin to maintain stem and progenitor cell fate by dictating the expression of specific classes of genes.

Gene losses, parallel evolution and heightened expression confer adaptations to dedicated cleaning behaviour.

BACKGROUND: Cleaning symbioses are captivating interspecific interactions in which a cleaner fish removes ectoparasites from its client, contributing to the health and diversity of natural fish communities and aquaculture systems. However, the genetic signatures underlying this specialized behaviour remain poorly explored. To shed light on this, we generated a high-quality chromosome-scale genome of the bluestreak cleaner wrasse Labroides dimidiatus, a dedicated cleaner with cleaning as primary feeding mechanism throughout its life. RESULTS: Compared with facultative and non-cleaner wrasses, L. dimidiatus was found with notable contractions in olfactory receptors implying their limited importance in dedicated cleaning. Instead, given its distinct tactile pre-conflict strategies, L. dimidiatus may rely more heavily on touch sensory perception, with heightened gene expression in the brain in anticipation of cleaning. Additionally, a reduction in NLR family CARD domain-containing protein 3 might enhance innate immunity of L. dimidiatus, probably assisting to reduce the impacts from parasite infections. In addition, convergent substitutions for a taste receptor and bone development genes across cleaners (L. dimidiatus and facultative cleaners) may provide them with evolved food discrimination abilities and jaw morphology that differentiate them from non-cleaners. Moreover, L. dimidiatus may exhibit specialized neural signal transductions for cleaning, as evidenced by positive selection in genes related to the glutamatergic synapse pathway. Interestingly, numerous glutamate receptors also demonstrated significantly higher expression in L. dimidiatus not engaged in cleaning, as compared to those involved in cleaning. Besides, apparent contractions in L. dimidiatus for protocadherins, which are responsible for neuronal development, may further promote specialized neural signal transductions in this species. CONCLUSIONS: This study reveals that L. dimidiatus harbours substantial losses in specific gene families, convergent evolutions across cleaners and a large-scale high gene expression in preparation for cleaning, allowing for adaptation to the dedicated cleaning behaviour.

Harnessing Oxylipins and Inflammation Modulation for Prevention and Treatment of Colorectal Cancer.

Colorectal cancer (CRC) is one of the most prevalent cancers worldwide, ranking as the third most malignant. The incidence of CRC has been increasing with time, and it is reported that Westernized diet and lifestyle play a significant role in its higher incidence and rapid progression. The intake of high amounts of omega-6 (n - 6) PUFAs and low levels of omega-3 (n - 3) PUFAs has an important role in chronic inflammation and cancer progression, which could be associated with the increase in CRC prevalence. Oxylipins generated from PUFAs are bioactive lipid mediators and have various functions, especially in inflammation and proliferation. Carcinogenesis is often a consequence of chronic inflammation, and evidence has shown the particular involvement of n - 6 PUFA arachidonic acid-derived oxylipins in CRC, which is further described in this review. A deeper understanding of the role and metabolism of PUFAs by their modifying enzymes, their pathways, and the corresponding oxylipins may allow us to identify new approaches to employ oxylipin-associated immunomodulation to enhance immunotherapy in cancer. This paper summarizes oxylipins identified in the context of the initiation, development, and metastasis of CRC. We further explore CRC chemo-prevention strategies that involve oxylipins as potential therapeutics.

High-efficiency focusing and imaging by dielectric kinoform zone plate lenses with soft X-rays.

With fast advances in enhancing the focusing/imaging resolution of Fresnel zone plate lenses toward sub-10 nm, low diffraction efficiency in connection with their rectangular zone shape still remains a big issue in both soft and hard X-ray microscopy. In hard X-ray optics, encouraging progress has recently been reported in our earlier attempts of high focusing efficiency by 3D kinoform shaped metallic zone plates, formed by greyscale electron beam lithography. This paper addresses our efforts towards high focusing/imaging efficiency by developing a novel dielectric kinoform zone plate lens for soft X-rays. The effects of the zone materials and zone shapes on the focusing/imaging quality were first theoretically investigated by a modified thin-grating-approximation method, revealing superior efficiencies of dielectric kinoform zone plates over rectangular ones in metals. Optical characterizations of replicated dielectric kinoform zone plates by greyscale electron beam lithography demonstrate a focusing efficiency of 15.5% with a resolution of 110 nm in the water window of X-rays. Apart from high efficiency, the novel kinoform zone plate lenses developed in this work exhibit significant advantages over conventional zone plates, i.e. simplified process, low cost and no need for a beamstop.

The Transcription Factor NIGT1.2 Modulates Both Phosphate Uptake and Nitrate Influx during Phosphate Starvation in Arabidopsis and Maize.

Phosphorus and nitrogen are essential macronutrients for plant growth and crop production. During phosphate (Pi) starvation, plants enhanced Pi but reduced nitrate (NO3 -) uptake capacity, and the mechanism is unclear. Here, we show that a GARP-type transcription factor NITRATE-INDUCIBLE, GARP-TYPE TRANSCRIPTIOANL REPRESSOR1.2 (NIGT1.2) coordinately modulates Pi and NO3 - uptake in response to Pi starvation. Overexpression of NIGT1.2 increased Pi uptake capacity but decreased NO3 - uptake capacity in Arabidopsis (Arabidopsis thaliana). Furthermore, the nigt1.1 nigt1.2 double mutant displayed reduced Pi uptake but enhanced NO3 - uptake under low-Pi stress. During Pi starvation, NIGT1.2 directly up-regulated the transcription of the Pi transporter genes PHOSPHATE TRANSPORTER1;1 (PHT1;1) and PHOSPHATE TRANSPORTER1;4 (PHT1;4) and down-regulated expression of NO3 - transporter gene NITRATE TRANSPORTER1.1 (NRT1.1) by binding to cis-elements in their promoters. Further genetic assays demonstrated that PHT1;1, PHT1;4, and NRT1.1 were genetically epistatic to NIGT1.2 We also identified similar regulatory pathway in maize (Zea mays). These data demonstrate that the transcription factor NIGT1.2 plays a central role in modulating low-Pi-dependent uptake of Pi and NO3 -, tending toward maintenance of the phosphorus to nitrogen balance in plants during Pi starvation.

CDK12 Is Necessary to Promote Epidermal Differentiation Through Transcription Elongation.

Proper differentiation of the epidermis is essential to prevent water loss and to protect the body from the outside environment. Perturbations in this process can lead to a variety of skin diseases that impacts 1 in 5 people. While transcription factors that control epidermal differentiation have been well characterized, other aspects of transcription control such as elongation are poorly understood. Here we show that of the two cyclin-dependent kinases (CDK12 and CDK13), that are known to regulate transcription elongation, only CDK12 is necessary for epidermal differentiation. Depletion of CDK12 led to loss of differentiation gene expression and absence of skin barrier formation in regenerated human epidermis. CDK12 binds to genes that code for differentiation promoting transcription factors (GRHL3, KLF4, and OVOL1) and is necessary for their elongation. CDK12 is necessary for elongation by promoting Ser2 phosphorylation on the C-terminal domain of RNA polymerase II and the stabilization of binding of the elongation factor SPT6 to target genes. Our results suggest that control of transcription elongation by CDK12 plays a prominent role in adult cell fate decisions.

Optimized Cationic Lipid-assisted Nanoparticle for Delivering CpG Oligodeoxynucleotides to Treat Hepatitis B Virus Infection.

PURPOSE: Hepatitis B virus (HBV) infection is such a global health problem that hundreds of millions of people are HBV carriers. Current anti-viral agents can inhibit HBV replication, but can hardly eradicate HBV. Cytosine-phosphate-guanosine (CpG) oligodeoxynucleotides (ODNs) are an adjuvant that can activate plasmacytoid dendritic cells (pDCs) and conventional dendritic cells (cDCs) to induce therapeutic immunity for HBV eradication. However, efficient delivery of CpG ODNs into pDCs and cDCs remains a challenge. In this study, we constructed a series of cationic lipid-assisted nanoparticles (CLANs) using different cationic lipids to screen an optimal nanoparticle for delivering CpG ODNs into pDCs and cDCs. METHODS: We constructed different CLANCpG using six cationic lipids and analyzed the cellular uptake of different CLANCpG by pDCs and cDCs in vitro and in vivo, and further analyzed the efficiency of different CLANCpG for activating pDCs and cDCs in both wild type mice and HBV-carrier mice. RESULTS: We found that CLAN fabricated with 1,2-Dioleoyl-3-trimethylammonium propane (DOTAP) showed the highest efficiency for delivering CpG ODNs into pDCs and cDCs, resulting in strong therapeutic immunity in HBV-carrier mice. By using CLANCpG as an immune adjuvant in combination with the injection of recombinant hepatitis B surface antigen (rHBsAg), HBV was successfully eradicated and the chronic liver inflammation in HBV-carrier mice was reduced. CONCLUSION: We screened an optimized CLAN fabricated with DOTAP for efficient delivery of CpG ODNs to pDCs and cDCs, which can act as a therapeutic vaccine adjuvant for treating HBV infection.

A compound Kinoform/Fresnel zone plate lens with 15 nm resolution and high efficiency in soft x-ray.

X-ray microscope as an important nanoprobing tool plays a prevailing role in nano-inspections of materials. Despite the fast advances of high resolution focusing/imaging reported, the efficiency of existing high-resolution zone plates is mostly around 5% in soft x-ray and rapidly goes down to 1%-2% when the resolution approaches 10 nm. It is well known that the rectangular zone shape, beamstop, limited height/width ratios, material absorption of light and structural defects are likely responsible for the limited efficiency. Although zone plates with Kinoform profile are supposed to be efficient, progress for achieving both high resolution (<30 nm) and high efficiency (>5%) have hardly been addressed in soft x-ray. In this work, we propose a compound Kinoform/Fresnel zone plate (CKZP) by combing a dielectric Kinoform zone plate with a 15 nm resolution zone plate. Greyscale electron beam lithography was applied to form the 3D Kinoform zone plate and atomic layer deposition was carried out to form the binary zone plate. Optical characterizations demonstrated 15 nm resolution focusing/imaging with over 7.8% efficiency in soft x-ray. The origin of the efficiency improvement behind the proposed compound lens is theoretically analyzed and discussed.

Design, synthesis and antifungal activity of novel amide derivatives containing a pyrrolidine moiety as potential succinate dehydrogenase inhibitors.

To discover new succinate dehydrogenase inhibitors (SDHI) fungicides, a series of amide derivatives containing a pyrrolidine moiety were designed and synthesized, and their antifungal activities were evaluated against Monilinia fructicola (M. fructicola), Rhizoctonia solani (R. solani), Fusarium graminearum schw (F. graminearum), Fusarium oxysporum (F. oxysporum), and Phytophthora infestans (P. infestans). Some compounds showed excellent antifungal activities against the five fungi. Among them, compound 6 showed broad-spectrum inhibitory activities. The EC50 of compound 6 against M. fructicola, R. solani, F. graminearum, F. oxysporum, and P. infestans were 2.13, 14.42, 1.69, 27.79, and 27.12 mg/L, respectively. In addition, compound 6 can effectively inhibit the spore germination of M. fructicola and has moderate damage to the cell membrane. Compound 6 can effectively inhibit succinate dehydrogenase (SDH) of M. fructicola, and can significantly increase the expression levels of SDHC and SDHD. Compound 6 can be used as a lead structure for developing new SDH inhibitors.

DDX6 Orchestrates Mammalian Progenitor Function through the mRNA Degradation and Translation Pathways.

In adult tissues, stem and progenitor cells must balance proliferation and differentiation to maintain homeostasis. How this is done is unclear. Here, we show that the DEAD box RNA helicase, DDX6 is necessary for maintaining adult progenitor cell function. DDX6 loss results in premature differentiation and decreased proliferation of epidermal progenitor cells. To maintain self-renewal, DDX6 associates with YBX1 to bind the stem loops found in the 3' UTRs of regulators of proliferation/self-renewal (CDK1, EZH2) and recruit them to EIF4E to facilitate their translation. To prevent premature differentiation of progenitor cells, DDX6 regulates the 5' UTR of differentiation inducing transcription factor, KLF4 and degrades its transcripts through association with mRNA degradation proteins. Our results demonstrate that progenitor function is maintained by DDX6 complexes through two distinct pathways that include the degradation of differentiation-inducing transcripts and by promoting the translation of self-renewal and proliferation mRNAs.

Dispersion-engineered broadband diffractive optical elements with multilayer subwavelength structures.

Diffractive optical elements (DOEs) play an important role in modern optical applications such as spectral and imaging systems, but it is challenging to balance the diffraction efficiency with the working bandwidth. The core issue is controlling the broadband dispersion of all phase units to achieve achromatic 2π-phase modulation in the broadband domain. Here, we demonstrate broadband DOEs utilizing multilayer subwavelength structures with different materials, making it possible to freely control the phase and phase dispersion of the structural units on a much larger scale than monolayer structures. The desired dispersion-control abilities arose due to a dispersion-cooperation mechanism and vertical mode-coupling effects between the top and bottom layers. An infrared design comprised of two vertically concatenated T i O 2 and Si nanoantennas separated by a S i O 2 dielectric spacer layer was demonstrated. It showed an average efficiency of over 70% in the three-octave bandwidth. This work shows enormous value for broadband optical systems with DOEs such as spectral imaging and augmented reality.

RAS-mediated suppression of PAR3 and its effects on SCC initiation and tissue architecture occur independently of hyperplasia.

Proper epithelial development and homeostasis depends on strict control of oriented cell division. Current evidence shows that this process is regulated by intrinsic polarity factors and external spatial cues. Owing to the lack of an appropriate model system that can recapitulate the architecture of the skin, deregulation of spindle orientation in human epithelial carcinoma has never been investigated. Here, using an inducible model of human squamous cell carcinoma (SCC), we demonstrate that RAS-dependent suppression of PAR3 (encoded by PARD3) accelerates epithelial disorganization during early tumorigenesis. Diminished PAR3 led to loss of E-cadherin-mediated cell adhesion, which in turn contributed to misoriented cell division. Pharmacological inhibition of the MAPK pathway downstream of RAS activation reversed the defects in PAR3 expression, E-cadherin-mediated cell adhesion and mitotic spindle orientation. Thus, temporal analysis of human neoplasia provides a powerful approach to study cellular and molecular transformations during early oncogenesis, which allowed identification of PAR3 as a critical regulator of tissue architecture during initial human SCC development.

Trapezoid-kinoform zone plate lens - a solution for efficient focusing in hard X-ray optics.

X-ray microscopes are powerful tools in the nano-inspection of materials owing to their ultra-high resolution at the molecular level. However, the focusing efficiency of binary zone plate lenses as key components in such probes is merely 5% in practice, hindering their application in advanced scientific research. Although kinoform zone plate lenses are in principle supposed to possess high efficiency beyond binary ones, little progress has been reported so far due to the shortage of both a theoretical calculation approach and greyscale lithography for generating fine three-dimensional (3D) kinoform zones of the lenses. This paper reports our theoretical work for a modified beam propagation method to compute the focusing performance and state-of-the-art 3D greyscale electron beam lithography for kinoform zone plate lenses. Three different zone shapes - binary, kinoform and top-flat kinoform (nicknamed the trapezoid-kinoform) - were compared both theoretically and experimentally. Theoretical calculations suggest, for the first time, that the trapezoid-kinoform zone plate gives rise to the highest focusing efficiency among the three lenses, which was proved by optical characterization of the fabricated lens with hard X-rays. As high as 40% of the focusing efficiency by Au trapezoid-kinoform lenses with resolution of 250 nm at 8 keV has been achieved, which is two times higher than that of binary zone plate lenses. The origin of the enhanced efficiency in the trapezoid-kinoform zone plate lens was explained by the joint contributions from both the refraction through the kinoform slope and the diffraction through the top flat part of the trapezoid-kinoform zone plate. Such a breakthrough in focusing efficiency sheds light on the further development of X-ray lenses with both high resolution and high efficiency.

Study of structural effects on the polarization characteristics of subwavelength metallic gratings in short infrared wavelengths.

Applications of subwavelength grating based-polarizers for polarimetric detections are being hindered due to the limited extinction ratio. In this work, the structural effect, including the line edge roughness (LER), of the gratings on the polarizing characteristics was studied by both numerical simulations using finite difference and time domain (FDTD) method and experiments, aiming to figure out the optimal grating profile for achieving high transmittance as well as high extinction ratio. Two different configurations of the gratings, one is dual layer Au lines and the other is parabolic shaped Al lines on structured spin-on-carbon (SOC) films were systematically studied and compared. Nanofabrication of the gratings by electron beam lithography without lift-off process were conducted and optical measurements of their polarization properties demonstrate superior performance of the developed polarizers. The origin of the structural effect was explained by the local surface plasmonic modes, existing in the nano-slits in metallic gratings, which is instructive for further enhancement of the polarization performance.

Infrared Characterization of Isotopic Analogues of Methanediol in Aqueous Solution.

Dissolved methanediol in aqueous solution has been treated as the precursor for the formation of atmospheric formic acid in multiphase environments. In this work, methanediol, CH2(OH)2, and its isotopic analogues, CH2(OD)2, CD2(OH)2, and CD2(OD)2, in aqueous solution were prepared by dissolving paraformaldehyde and deuterium-substituted paraformaldehyde powders in H2O and D2O under reflux. Their infrared absorption contours of formaldehyde solutions at concentrations of <1 wt % are not dependent on the concentration, mainly referring to the characteristics of the monomeric configuration, and can be categorized into two parts. At wavenumbers >2000 cm-1, broad bands of moderate strengths were ascribed to the stretching modes of two OH or OD groups, observed at 3000-3700 and 2050-2750 cm-1, respectively. At wavenumbers of 950-1200 cm-1, the isotopic analogues of methanediols composed of CH2 moieties are featured with a singlet strong band at ca. 1030 cm-1, mainly attributed to the O-C-O stretching modes; the isotopic methanediols containing CD2 moieties manifested two intense bands at ca. 1100 and 980 cm-1, majorly enveloping the CD2 deformation and O-C-O stretching modes. The aforementioned spectral features were assigned on the basis of density functional theory, ωB97XD, with the basis set aug-cc-pVTZ and the solvent effect using the conductor-like polarizable continuum model. In addition, the predicted energetics suggested that the trans-methanediol is more stable than the cis- conformer by ca. 0.62 kcal mol-1 and majorly contributes to the infrared features. At higher concentrations of CH2(OH)2, extra bands at 920 and 1104 cm-1 appeared and were attributed to the C-O-C stretching modes of the dimeric/polymeric methanediol; that is, HO(CH2O)nH, n ≥ 2. These infrared characterizations of the isotopic analogues of methanediols provided suitable detection windows in the relevant atmospheric and aerosol reactions in the laboratory studies.

Spectral filter in short-wave infrareds based on a metasurface on silicon-on-insulator with polarizing bandpass.

Spectral filters with polarimetric character in short-wave infrareds are urgently needed because of their broad applications in optic-fiber communications, polarimetric detections, and imaging. Based on our earlier progress in developing polarimetric devices in infrared wavelengths, in this work, a plasmonic-metasurface-based polarization-dependency multi-channel narrowband filter in short-wave infrareds was developed. To meet the requirement by the developing trend of polarimetric detection/spectral imaging in short-wave infrareds, a resonant cavity in the form of the Au hat/elliptical Si/SiO2 pillars/Au layer as the filters was proposed. Numerical simulations by finite-difference time-domain (FDTD) show resonant and polarized transmissions of the designed devices to infrared light in short wavelengths, and the peak positions are relevant to the structural dimensions. Optical characteristics of the filters, fabricated by electron beam lithography/dry-etch technique, agree well with the simulated behavior. To enhance the transmission efficiency to the applicable level, nanoprocessing of the filters still needs to be optimized. Nevertheless, the progress reported is promising for this new type of spectral filters based on modern metasurfaces.

Correction: Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development.

[This corrects the article DOI: 10.1371/journal.pgen.1005833.].

Recent research progress and outlook in agricultural chemical discovery based on quinazoline scaffold.

The discovery of new scaffolds and targets for pesticides is still a huge challenge facing the sustainable development of modern agriculture. In recent years, quinazoline derivatives have achieved great progress in drug discovery and have attracted great attention. Quinazoline is a unique bicyclic scaffold with a variety of biological activities, which increases the possibilities and flexibility of structural modification, showing enormous appeal in the discovery of new pesticides. Therefore, the agricultural biological activities, structure-activity relationships (SAR), and mechanism of action of quinazoline derivatives in the past decade were reviewed systematically, with emphasis on SAR and mechanism. Then, we prospected the application of the quinazoline scaffold as a special structure in agricultural chemical discovery, hoping to provide new ideas for the rational design and mechanism of novel quinazoline agricultural chemicals in the future.