Variation in Bombyx mori immune response against fungal pathogen Beauveria bassiana with variability in cell wall ß-1,3-glucan.

Entomopathogenic fungi are protected by a cell wall with dynamic structure for adapting to various environmental conditions. ß-1,3-Glucan recognition proteins activate the innate immune system of insects by recognizing surface molecules of fungi. However, the associations between pathogenicity and the different components of entomopathogenic fungal cell walls remain unclear. Three Beauveria bassiana strains were selected that have significantly differing virulence against Bombyx mori. The molecular mechanisms underlying the immune response in B. mori were investigated using RNA sequencing, which revealed differences in the immune response to different B. bassiana strains at 12 h post-infection. Immunofluorescence assays revealed that ß-1,3-glucan content had an opposite trend to that of fungal virulence. ß-1,3-Glucan injection upregulated BmßGRP4 expression and significantly reduced the virulence of the high-virulence strain but not that of the medium-virulence or low-virulence strains. BmßGRP4 silencing in B. mori with RNA interference resulted in the opposite virulence pattern, indicating that the virulence of B. bassiana was affected by the cell walls' content of ß-1,3-glucan, which could be recognized by BmßGRP4. Furthermore, interference with the gene CnA (calcineurin catalytic A subunit) involved in ß-1,3-glucan synthesis eliminated differences in virulence between B. bassiana strains. These results indicate that strains of a single species of pathogenic fungi that have differing cell wall components are recognized differently by the innate immune system of B. mori.

MicroRNA-like RNA Functions Are Required for the Biosynthesis of Active Compounds in the Medicinal Fungus Sanghuangporus vaninii.

miRNA-like RNAs (milRNAs) have been recognized as sequence-specific regulators of posttranscriptional regulation of gene expression in eukaryotes. However, the functions of hundreds of fungal milRNAs in the biosynthesis of metabolic components are obscure. Sanghuangporus produces diverse bioactive compounds and is widely used in Asian countries. Here, genes encoding two Dicers, four Argonautes, and four RdRPs were identified and characterized in Sanghuangporus vanini. Due to the lack of an efficient gene manipulation system, the efficacy of spray-induced gene silencing (SIGS) was determined in S. vanini, which showed efficient double-stranded RNA (dsRNA) uptake and gene silencing efficiency. SIGS-mediated gene knockdown showed that SVRDRP-3, SVRDRP-4, SVDICER-1, and SVDICER-2 were critical for mycelial biomass, flavonoid, triterpenoid, and polysaccharide production. Illumina deep sequencing was performed to characterize the milRNAs from S. vanini mycelium and fruiting body. A total of 31 milRNAs were identified, out of which, SvmilR10, SvmilR17, and SvmilR33 were Svrdrp-4- and Svdicer-1-dependent milRNAs. Importantly, SIGS-mediated overexpression of SvmilR10 and SvmilR33 resulted in significant changes in the yields of flavonoids, triterpenoids, and polysaccharides. Further analysis showed that these milRNA target genes encoding the retrotransposon-derived protein PEG1 and histone-lysine N-methyltransferase were potentially downregulated in the milRNA overexpressing strain. Our results revealed that S. vanini has high external dsRNA and small RNA uptake efficiency and that milRNAs may play crucial regulatory roles in the biosynthesis of bioactive compounds. IMPORTANCE Fungi can take up environmental RNA that can silence fungal genes with RNA interference, which prompts the development of SIGS. Efficient dsRNA and milRNA uptake in S. vanini, successful dsRNA-targeted gene block, and the increase in intracellular miRNA abundance showed that SIGS technology is an effective and powerful tool for the functional dissection of fungal genes and millRNAs. We found that the RdRP, Dicer, and Argonaute genes are critical for mycelial biomass and bioactive compound production. Our study also demonstrated that overexpressed SVRDRP-4- and SVDICER-1-dependent milRNAs (SvmilR10 and SvmilR33) led to significant changes in the yields of the three active compounds. This study not only provides the first report on SIGS-based gene and milRNA function exploration, but also provides a theoretical platform for exploration of the functions of milRNAs involved in biosynthesis of metabolic compounds in fungi.

Functional Analysis of Keto-Acid Reductoisomerase ILVC in the Entomopathogenic Fungus Metarhizium robertsii.

Ketol-acid reductoisomerase (ILVC) is the second enzyme in the branched-chain amino acid (BCAA) biosynthesis, which regulates many physiological activities in a variety of organisms from bacteria to fungi and plants. In this work, function mechanisms of ILVC in Metarhizium robertsii Metchnikoff (Hypocreales: Clavicipitaceae) were explored with site-directed mutagenesis, reductase activity assays and transcriptomics analysis. The reductase activity assays showed that ILVC from phytopathogenic fungi exhibited significantly higher activities than those from entomopathogenic fungi but lower than those from yeast. Site-directed mutagenesis and enzymatic activities of MrILVC with different active-site mutants (Arg-113, Ser-118, Asp-152, Asp-260, and Glu-264) confirmed that active sites of MrILVC are conserved with plant and bacterial ILVCs. Deleting MrilvC causes the complete failures of vegetative growth and conidial germination, feeding with branched-chain amino acids (BCAAs) recovers the fungal growth but not conidial germination, while both characteristics are restored when supplemented with yeast extract. Compared to ΔMrilvC cultured in czapek agar (CZA), plenty of genes involved in the biosynthesis of antibiotics and amino acids were up- or down-regulated in the wild type or ΔMrilvC feeding with either BCAAs or yeast extract. Further analysis showed some genes, such as catalase A, participate in mycelial growth and conidial germination was down-regulated in ΔMrilvC from CZA, revealing that MrILVC might control the fungal development by gene regulation and BCAAs or yeast extract could play partial roles of MrILVC. This study will advance our understanding of ILVC function mechanisms in fungi.

High quality N-polar GaN films grown with varied V/III ratios by metal-organic vapor phase epitaxy.

We studied the growths and characterizations of N-polar GaN films grown with constant and varied V/III ratios in high-temperature (HT) GaN growth on offcut c-plane sapphire substrates by metal-organic vapor phase epitaxy. It is found that growth with a constantly low V/III ratio resulted in a high crystallinity but a rough surface and a high oxygen concentration, whereas growth with a high V/III ratio led to a smooth surface but a high carbon concentration and a degraded crystallinity. The overall quality of the N-polar GaN epilayer cannot be effectively improved simply by tuning the V/III ratio. The growth with varied V/III ratios was conducted by lowering the V/III ratio in the initial HT-GaN growth and keeping the V/III ratio constantly high in the subsequent growth. Such a change of V/III ratio resulted in a 3D-to-2D like growth mode transition during the early stage of HT-GaN growth which helped reduce threading dislocations and suppress impurity incorporation. By optimizing the nucleation temperature and the thickness of the initial low-V/III-ratio layer, the minimum full-widths at half-maximum of (002̄)/(102̄) rocking curves obtained were 288/350 arcsec and the oxygen concentration was reduced significantly from 1.6 × 1018 cm-3 to 3.7 × 1017 cm-3 while keeping a hillock-free smooth surface morphology. The overall quality of the N-polar GaN films was considerably improved. We believe that this simple, yet effective growth technique has great application prospects for high-performance N-polar GaN-based electron devices.

Pavlovian conditioning demonstrated with neuromorphic memristive devices.

Pavlovian conditioning, a classical case of associative learning in a biological brain, is demonstrated using the Ni/Nb-SrTiO3/Ti memristive device with intrinsic forgetting properties in the framework of the asymmetric spike-timing-dependent plasticity of synapses. Three basic features of the Pavlovian conditioning, namely, acquisition, extinction and recovery, are implemented in detail. The effects of the temporal relation between conditioned and unconditioned stimuli as well as the time interval between individual training trials on the Pavlovian conditioning are investigated. The resulting change of the response strength, the number of training trials necessary for acquisition and the number of extinction trials are illustrated. This work clearly demonstrates the hardware implementation of the brain function of the associative learning.

Mimicking the brain functions of learning, forgetting and explicit/implicit memories with SrTiO3-based memristive devices.

To implement the complex brain functions of learning, forgetting and memory in a single electronic device is very advantageous for realizing artificial intelligence. As a proof of concept, memristive devices with a simple structure of Ni/Nb-SrTiO3/Ti were investigated in this work. The functions of learning, forgetting and memory were successfully mimicked using the memristive devices, and the "time-saving" effect of implicit memory was also demonstrated. The physics behind the brain functions is simply the modulation of the Schottky barrier at the Ni/SrTiO3 interface. The realization of various psychological functions in a single device simplifies the construction of the artificial neural network and facilitates the advent of artificial intelligence.

Pt/WO3/FTO memristive devices with recoverable pseudo-electroforming for time-delay switches in neuromorphic computing.

A recoverable pseudo-electroforming process was discovered in Pt/WO3/FTO devices. Unlike conventional electroforming, which is usually destructive, pseudo-electroforming can be recovered when the electrical stimulation is removed. Furthermore, the time-dependent recovery process can be tuned by diverse voltage pulses applied in pseudo-electroforming; therefore, the device can be used as a time-delay switch in memristor-based neuromorphic networks. This "volatile" electroforming process can be attributed to the high oxygen vacancy concentration in the fluorine-doped tin oxide (FTO) bottom electrode, which acts as a non-blocking electrode in the resistive switching.

Revival of "dead" memristive devices: case of WO3-x.

Inappropriate operation could make a memristive device "dead" and cause the loss of resistive switching performance. In this study, the revival of "dead" devices was investigated in the case of WO3-x-based memristive devices. It is believed that inappropriate operation with a high-voltage pulse creates an ordered structure of oxygen vacancies and such an ordered structure makes the normal reset process fail. By precisely controlled voltage sweeping at certain compliance currents, a "dead" device can be revived. The revival operation disrupts the ordered structure by Joule heating and recovers Schottky-like barrier modulation-based switching.

Synaptic Metaplasticity Realized in Oxide Memristive Devices.

Metaplasticity, a higher order of synaptic plasticity, as well as a key issue in neuroscience, is realized with artificial synapses based on a WO3 thin film, and the activity-dependent metaplastic responses of the artificial synapses, such as spike-timing-dependent plasticity, are systematically investigated. This work has significant implications in neuromorphic computation.

Effects of cognitive rehabilitation training on schizophrenia: 2 years of follow-up.

OBJECTIVE: Schizophrenia is a mental disorder and characterized by abnormal social behavior and failure to recognize what is real. The current study was to explore the long-term effects of cognitive rehabilitation training on schizophrenia. METHODS: Eighty six cases of hospitalized patients with schizophrenia were randomly divided into study group and control group. The relapse and employment (attending school) rates were used as indicators to assess the treatment effect. All patients were followed up by 2 years. Kaplan-Meier survival analysis was conducted with relapse and employment (attending school) rates. RESULTS: The rates of relapse in the study group and the control group were 18% and 41%, and relapse free survival time was 22.22 months and 18.55 months; the rates of employment (attending school) were 64% and 43%, and not employment (attending school) time were 10.68 months and 15.74 months, respectively. There was significant difference between the two groups (P<0.05). CONCLUSIONS: We found that the cognitive rehabilitation training could significantly reduce schizophrenic relapse rate, prolong the time of patients without relapse, improve the employment (attending school) rate, and shorten the discharged time, which is a powerful treatment method to improve social competence in schizophrenia patients.

The role of Schottky barrier in the resistive switching of SrTiO3: direct experimental evidence.

Single crystalline SrTiO3 doped with 0.1 wt% Nb was used as a model system to evaluate the role of the Schottky barrier in the resistive switching of perovskites. The Ti bottom electrode formed an ohmic contact in the Ni/Nb:SrTiO3/Ti stack, whereas the Ni top electrode created a strong Schottky barrier, which was reflected in a huge semi-circle in the impedance spectrum of the stack. Bipolar switching was achieved in the voltage range of -4 to 4 V for the stack, two clear resistance states were created by electric pulses, and the Schottky barrier heights corresponding to the high/low resistance states were experimentally determined. A direct relationship between the resistance state and the Schottky barrier height was thus established.

Acoustic grating fringe projector for high-speed and high-precision three-dimensional shape measurements.

A new acoustic grating fringe projector (AGFP) was developed for high-speed and high-precision 3D measurement. A new acoustic grating fringe projection theory is also proposed to describe the optical system. The AGFP instrument can adjust the spatial phase and period of fringes with unprecedented speed and accuracy. Using rf power proportional-integral-derivative (PID) control and CCD synchronous control, we obtain fringes with fine sinusoidal characteristics and realize high-speed acquisition of image data. Using the device, we obtained a precise phase map for a 3D profile. In addition, the AGFP can work in running fringe mode, which could be applied in other measurement fields.