MicroRNA-542-3p targets Pten to inhibit the myoblasts proliferation but suppresses myogenic differentiation independent of targeted Pten.

BACKGROUND: Non-coding RNA is a key epigenetic regulation factor during skeletal muscle development and postnatal growth, and miR-542-3p was reported to be conserved and highly expressed in the skeletal muscle among different species. However, its exact functions in the proliferation of muscle stem cells and myogenesis remain to be determined. METHODS: Transfection of proliferative and differentiated C2C12 cells used miR-542-3p mimic and inhibitor. RT-qPCR, EdU staining, immunofluorescence staining, cell counting kit 8 (CCK-8), and Western blot were used to evaluate the proliferation and myogenic differentiation caused by miR-542-3p. The dual luciferase reporter analysis and rescued experiment of the target gene were used to reveal the molecular mechanism. RESULTS: The data shows overexpression of miR-542-3p downregulation of mRNA and protein levels of proliferation marker genes, reduction of EdU+ cells, and cellular vitality. Additionally, knocking it down promoted the aforementioned phenotypes. For differentiation, the miR-542-3p gain-of-function reduced both mRNA and protein levels of myogenic genes, including MYOG, MYOD1, et al. Furthermore, immunofluorescence staining immunized by MYHC antibody showed that the myotube number, fluorescence intensity, differentiation index, and myotube fusion index all decreased in the miR-542-3p mimic group, compared with the control group. Conversely, these phenotypes exhibited an increased trend in the miR-542-3p inhibitor group. Mechanistically, phosphatase and tensin homolog (Pten) was identified as the bona fide target gene of miR-542-3p by dual luciferase reporter gene assay, si-Pten combined with miR-542-3p inhibitor treatments totally rescued the promotion of proliferation by loss-function of miR-542-3p. CONCLUSIONS: This study indicates that miR-542-3p inhibits the proliferation and differentiation of myoblast and Pten is a dependent target gene of miR-542-3p in myoblast proliferation, but not in differentiation.

CRISPR/Cas9-mediated knockout of the abdominal-B homeotic gene in the global pest, fall armyworm (Spodoptera frugiperda).

The Homeotic complex (Hox) genes play a crucial role in determining segment identity and appendage morphology in bilaterian animals along the antero-posterior axis. Recent studies have expanded to agricultural pests such as fall armyworm (FAW), scientifically known as Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae), which significantly threatens global agricultural productivity. However, the specific role of the hox gene Sfabd-B in FAW remains unexplored. This research investigates the spatial and temporal expression patterns of Sfabd-B in various tissues at different developmental stages using quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, we explored the potential function of the Sfabd-B gene located in the FAW genome using CRISPR/Cas9 technology. The larval mutant phenotypes can be classified into three subgroups as compared with wild-type individuals, that is, an excess of pedis in the posterior abdomen, deficient pedis due to segmental fusion and deviations in the posterior abdominal segments. Importantly, significant differences in mutant phenotypes between male and female individuals were also evident during the pupal and adult phases. Notably, both the decapentaplegic (dpp) and cuticular protein 12 (cp 12) genes displayed a substantial marked decrease in expression levels in the copulatory organ of male mutants and the ovipositor of female mutants compared with the wild type. These findings highlight the importance of Sfabd-B in genital tract patterning, providing a potential target for improving genetic control.

Inhibitory activity and antioomycete mechanism of citral against Phytophthora capsici.

The natural terpenoid citral has antifungal activity against multiple fungi, but its bioactivity against oomycetes is unclear. Therefore, this study investigated the antioomycete activity and mechanism of citral against Phytophthora capsici, a highly destructive invasive oomycete. Results showed that citral not only had a great inhibition on the mycelial growth of P. capsici (EC50 = 94.15 mg/L), but also had a significant inhibition on multiple spores, such as sporangia formation, zoospore discharge and zoospore germination. Citral at 4000 mg/L exhibited favorable protective (73.33%) and curative efficacy (55.11%) against pepper Phytophthora blight. Citral significantly damaged the hyphal morphology, disrupted the cell membrane integrity, increased the permeability of cell membrane, and increased the glycerol content in P. capsici. A total of 250 upregulated and 288 downregulated proteins were identified in iTRAQ-based quantitative proteomic analysis. Downregulated proteins were mostly enriched in pathways of ABC transporters, cyanoamino acid metabolism and starch and sucrose metabolism, suggesting an inhibition of citral on transmembrane transporter (e.g., ABC transporters) and pathogenicity (e.g., ß-glucosidases) proteins. Upregulated proteins were enriched in biosynthesis of unsaturated fatty acids, pyruvate metabolism and glycolysis/gluconeogenesis, suggesting an activation of citral on energy generation proteins, including acyl-CoA oxidase, D-lactate dehydrogenase, pyruvate kinase, acetyl-CoA synthetase and phosphoenolpyruvate carboxykinase. Biochemical and iTRAQ analysis suggested that cell membrane may be the target of citral in P. capsici.

Accumulation and metabolism of pyroxasulfone in tomato seedlings.

Pyroxasulfone (PYS) is an isoxazole herbicide favored for its high activity. However, the metabolic mechanism of PYS in tomato plants and the response mechanism of tomato to PYS are still lacking. In this study, it was found that tomato seedlings had a strong ability to absorb and translocate PYS from roots to shoots. The highest accumulation of PYS was in the apex tissue of the tomato shoots. Using UPLC-MS/MS, five metabolites of PYS were detected and identified in tomato plants, and their relative contents in different parts of tomato plants varied greatly. The serine conjugate, DMIT [5, 5-dimethyl-4, 5-dihydroisoxazole-3-thiol (DMIT)] &Ser, was the most abundant metabolites of PYS in tomato plants. In tomato plants, the conjugation of thiol-containing metabolic intermediates of PYS to serine may mimic the cystathionine ß-synthase-catalyzed condensation of serine and homocysteine (in the pathway sly00260 sourced from KEGG database). This study ground breakingly proposed that serine may play an important role in plant metabolism of PYS and fluensulfone (whose molecular structure is similar to PYS). PYS and atrazine (whose toxicity profile is similar to PYS but not conjugate with serine) produced different regulatory outcomes for endogenous compounds in the pathway sly00260. Differential metabolites in tomato leaves exposed to PYS compared with the control, including amino acids, phosphates, and flavonoids, may play important roles in tomato response to PYS stress. This study provides inspiration for the biotransformation of sulfonyl-containing pesticides, antibiotics and other compounds in plants.

Bioactivity and Control Efficacy of Benzovindiflupyr Against Athelia rolfsii in China.

Athelia rolfsii is a devastating soilborne pathogen that causes stem rot of peanut and severely restricts peanut production. The new generation of succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr has been registered in the United States and Brazil for managing multiple plant diseases. However, it is not registered in China to control peanut stem rot. In this study, 246 isolates from major peanut production areas in Shandong, Henan, and Hebei Provinces of China were used to determine the baseline sensitivity of A. rolfsii to benzovindiflupyr. The frequency of EC50 values of benzovindiflupyr was unimodally distributed with an average EC50 of 0.12 ± 0.05 mg/liter and a range of 0.01 to 0.57 mg/liter. Benzovindiflupyr can also strongly inhibit the germination of sclerotia, with an average EC50 of 2.38 ± 1.04 mg/liter (n = 23). In addition, benzovindiflupyr exhibited great in vivo efficacy against A. rolfsii; the protective or curative efficacy (89.87%, 20.39%) of benzovindiflupyr at a concentration of 50 mg/liter was equivalent to that of the control fungicide thifluzamide at 100 mg/liter (86.39%, 16.21%). At the same concentration (e.g., 100 mg/liter), the protective efficacy (93.99%) of benzovindiflupyr was more than twice as high as the curative efficacy (45.07%). A positive correlation existed between benzovindiflupyr and isopyrazam or mefentrifluconazole, which possibly resulted from similar chemical structures or damage to the cell membrane. Our findings provide valuable information for the application of benzovindiflupyr, and the established baseline sensitivity could facilitate the monitoring and assessment of benzovindiflupyr resistance risk.

Selection and Validation of Reference Genes for Reverse-Transcription Quantitative PCR Analysis in Sclerotium rolfsii.

Reference genes are important for the accuracy of gene expression profiles using reverse-transcription quantitative PCR (RT-qPCR). However, there are no available reference genes reported for Sclerotium rolfsii; it actually has a pretty diverse and wide host range. In this study, seven candidate reference genes (UBC, ß-TUB, 28S, 18S, PGK, EF1α and GAPDH) were validated for their expression stability in S. rolfsii under conditions of different developmental stages, populations, fungicide treatments, photoperiods and pHs. Four algorithm programs (geNorm, Normfinder, Bestkeeper and ΔCt) were used to evaluate the gene expression stability, and RefFinder was used to integrate the ranking results of four programs. Two reference genes were recommended by RefFinder for RT-qPCR normalization in S. rolfsii. The suitable reference genes were GAPDH and UBC across developmental stages, PGK and UBC across populations, GAPDH and PGK across fungicide treatments, EF1α and PGK across photoperiods, ß-TUB and EF1α across pHs and PGK and GAPDH across all samples. Four target genes (atrB, PacC, WC1 and CAT) were selected for the validation of the suitability of selected reference genes. However, using one or two reference genes in combination to normalize the expression of target genes showed no significant difference in S. rolfsii. In short, this study provided reliable reference genes for studying the expression and function of genes in S. rolfsii.

Baseline sensitivity and bioactivity of tetramycin against Sclerotium rolfsii isolates in Huanghuai peanut-growing region of China.

Peanut stem rot caused by Sclerotium rolfsii is a serious soil-borne disease and poses a threat to the peanut production. The antibiotic fungicide tetramycin has a broad antifungal spectrum against multiple pathogens and possess low environmental risks. In current study, a total of 250 isolates collected from Huanghuai peanut-growing region of China (Henan, Shandong and Hebei Province) were used to establish the baseline sensitivity of S. rolfsii to tetramycin. The baseline sensitivity curve was unimodal and distributed from 0.01 to 0.36 mg/L, with a mean EC50 (50% effective concentration) value of 0.11 ± 0.06 mg/L. Tetramycin also had strong inhibitory activity on the formation and germination of sclerotia. There was no significant correlation of S. rolfsii sensitivity to tetramycin and other commonly used SDHI (succinate dehydrogenase inhibitor), QoI (quinone outside respiration inhibitor) and DMI (demethylation inhibitor) fungicides. Moreover, tetramycin significantly increased the cell membrane permeability and reduced the oxalate acid content. Greenhouse experiments showed that tetramycin has both protective and curative efficacy against S. rolfsii, while protective efficacy was higher than curative efficacy. Anyhow, the bioactivity of tetramycin is similar (curative efficacy) or higher (protective efficacy) than the control fungicide validamycin. In terms of application method, root drench may be more suitable for tetramycin than spraying, because root drench of tetramycin obtained a higher efficacy. These results indicated that tetramycin may be a potential alternative fungicide for the efficient control of peanut stem rot.

iTRAQ proteomic analysis of the inhibitory effect of 1,6-O,O-diacetylbritannilactone on the plant pathogenic oomycete Phytophthora capsici.

Phytophthora capsici is a highly destructive oomycete of vegetables; its management is challenging due to its broad host range, rapid dispersion, resilient spores and severe fungicide resistance. Identifying an effective alternative fungicide is important for the control of P. capsici. 1,6-O,O-diacetylbritannilactone (ABLOO), one of the secondary metabolites of Inula Britannica, showed a favorable inhibitory activity against P. capsici at different developmental stages, with a sensitivity order as follows: sporangia formation (30.45 mg/L) > zoospore discharge (77.69 mg/L) > mycelial growth (93.18 mg/L) > cystospore germination (591.48 mg/L). To investigate the mode of action of ABLOO in P. capsici, iTRAQ-based quantitative proteomic analysis was performed by comparing the expression levels of proteins in the control and ABLOO-treated (400 mg/L, inhibition rate of 80%) mycelial groups. A total of 65 downregulated and 75 upregulated proteins were identified in the proteomic analysis. Functional enrichment analyses showed that proteins with transmembrane transport activity were significantly inhibited, while proteins involved in energy production were significantly increased, including proteins involved in ubiquinone and other terpenoid-quinone biosynthesis, oxidative phosphorylation, and glycolysis/gluconeogenesis. The morphological results indicated that ABLOO treatment could decrease the thickness of the cell walls of P. capsici mycelia. Correspondingly, biochemical results showed that ABLOO treatment reduced the ß-1,3-glucan contents (the key component of the cell wall of P. capsici) and increased the cell membrane permeability of P. capsici. ABLOO may exhibit antioomycete activity by destroying the cell membrane of P. capsici. This study provides new evidence regarding the inhibitory mechanisms of ABLOO against P. capsici.

A Joint Low-Rank and Sparse Method for Reference Signal Purification in DTMB-Based Passive Bistatic Radar.

In a digital terrestrial multimedia broadcasting (DTMB)-based passive bistatic radar (PBR) system, the received reference signal often suffers from serious multipath effect, which decreases the detection ability of low-observable targets in urban environments. In order to improve the target detection performance, a novel reference signal purification method based on the low-rank and sparse feature is proposed in this paper. Specifically, this method firstly performs synchronization operations to the received reference signal and thus obtains the corresponding pseudo-noise (PN) sequences. Then, by innovatively exploiting the inherent low-rank structure of DTMB signals, the noise component in PN sequences is reduced. After that, a temporal correlation (TC)-based adaptive orthogonal matching pursuit (OMP) method, i.e., TC-AOMP, is performed to acquire the reliable channel estimation, whereby the previous noise-reduced PN sequences and a new halting criterion are utilized to improve channel estimation accuracy. Finally, the purification reference signal is obtained via equalization operation. The advantage of the proposed method is that it can obtain superior channel estimation performance and is more efficient compared to existing methods. Numerical and experimental results collected from the DTMB-based PBR system are presented to demonstrate the effectiveness of the proposed method.

Quantitative structure-chromatographic retention relationship of synthesized peptides (HGRFG, NPNPT) and their derivatives.

Carapax Trionycis extract peptides (HGRFG, NPNPT) are able to protect against CCl4-induced liver fibrosis. Therefore, this study applies to deal with chromatographic lipophilicity determination of synthesized peptides (HGRFG, NPNPT) and their derivatives using reversed-phase high performance liquid chromatography (RP-HPLC) combined with methanol-water mobile phase and two reversed-phase chromatographic columns (COSMOISL 5C18-MS-II and SHIMADZU-C18). The chromatographic lipophilicity of the analyzed compounds was expressed as logkw constant and correlated with lipophilicity descriptors. Quantitative structure-retention relationships (QSRR) analysis was performed to imitate chromatographic lipophilicity behavior using molecular descriptors. Modeling was performed using linear regression (LR) and multiple linear regression (MLR) methods with the help of principal component analysis (PCA) and hierarchical cluster analysis (HCA). The most influential molecular descriptors were lipophilicity descriptors, which are important for molecules ability to pass through biological membranes. All established QSRR models were statistically validated by standards, cross- and external validation parameters. According to these statistical validation parameters, MLR models (R2 > 0.856) were better for chromatographic lipophilicity prediction of peptide compounds. It can be concluded that chromatographic systems with COSMOISL 5C18-MS-II column were better for modeling of logkw than systems with SHIMADZU-C18 column. Modeling was performed in order to obtain lipophilicity profiles of investigated compounds as future drug candidates.

Characterization of Limonoids Isolated from the Fruits of Melia toosendan and Their Antifeedant Activity against Pieris rapae.

The fruits of Melia toosendan Sieb. et Zucc. (Meliaceae) are a source of bioactive limonoids that can be used as effective pesticides. In this study, two novel limonoids, 6-acetylsendanal and 6-ketocinamodiol, were isolated together with fourteen known compounds, namely four protolimonoids, six trichilin-class limonoids, and four C-seco limonoids. The structures of the new compounds were determined by extensive spectroscopic analyses (HR-ESI-MS, UV, IR, 1D and 2D NMR). The bioassay results revealed that eleven of the extracted limonoids exhibited interesting antifeedant activities against the larvae of Pieris rapae with AFC50 values in the range of 0.11-1.79 mm. Particularly, mesendanin H, with an AFC50 value of 0.11 mm, exhibited a higher activity than the positive control toosendanin. Information on new bioactive limonoids may provide further insight into M. toosendan as a source of bioactive components.

Insights into the phylogeny of Hemiptera from increased mitogenomic taxon sampling.

Although reconstruction of the phylogeny of Hemiptera has progressed tremendously over the past two decades, some higher-level relationships remain poorly resolved. Here, we investigated the Hemiptera higher-level relationships using full mitochondrial genome data from 357 ingroup species, representing the most comprehensive sampling yet undertaken for reconstructing the phylogeny of this group. In this study, 92 mitochondrial genomes were newly determined. Various data treatment methods and substitution models were applied to tree reconstructions. Effects of compositional heterogeneity, rate heterogeneity, model adequacy and taxon sampling on support values and topological stability were explored. Phylogenetic analyses (1) confirmed the monophyly of Hemiptera under site-heterogeneous model, (2) placed Sternorrhyncha as sister to all other Hemiptera, (3) recovered Coccoidea as the sister taxon of Aphidoidea, followed successively by Aleyrodoidea and Psylloidea, and (4) indicated that the grouping of Coleorrhyncha and Fulgoromorpha was the result of long-branch attraction effect.

Plant volatiles mediated the orientation preference of slugs to different plant species.

BACKGROUND: Slugs mechanically damage plant leaves, resulting in significant economic losses. However, there are limited cost-efficient strategies available in slug management. By studying how slugs utilize plant volatiles to locate host plants, we can gain insights into the design of attractants and repellents. RESULTS: Bioassay results suggest slugs (Agriolimax agrestis) prefer to orientate to lettuce (Lactuca sativa), cabbage (Brassica oleracea L.), and young tobacco seedlings, compared with old tobacco seedlings. We analyzed the volatomics of lettuce, cabbage, young and old tobacco seedlings. 2-(2-butoxyethoxy)-ethanol acetate (2EA) had high abundance while nonanal, decanal, and ß-cylocitral had relatively low content in volatiles. Old tobacco seedlings released significantly more hexanal but fewer 1,4-dihydro-4-oxopyridazine (DO). In olfactory tests, hexanal, nonanal, decanal, and ß-cylocitral showed strong repellency to slugs, while DO at a dose of 500 ng/µL and 2EA at a dose of 1% were attractive to slugs. The two alkanes, hexadecane and heptadecane, had no effect on slug orientating to host plants. DO and 2EA can thus alleviate the repellency of hexanal, nonanal, decanal and ß-cylocitral. CONCLUSION: The high emission of hexanal in old tobacco seedlings helps repel slugs, while 2EA and DO attract slugs to lettuce and cabbage. These findings suggest that these chemicals can be utilized in the design of repellents and attractants, and contribute to constructing a push-pull system for slug control. © 2023 Society of Chemical Industry.

Phase-Separated Nano-Antibiotics Enhanced Survival in Multidrug-Resistant Escherichia coli Sepsis by Precise Periplasmic EcDsbA Targeting.

Disulfide bond (Dsb) proteins, especially DsbA, represent a promising but as-yet-unrealized target in combating multidrug-resistant (MDR) bacteria because their precise subcellular targeting through multibarrier remains a significant challenge. Here, a novel heterogenization-phase-separated nano-antibiotics (NCefoTs) is proposed, through the co-assembly of enzyme-inhibiting lipopeptides (ELp component), membrane-recognizing and disrupting lipopeptides (MLp component), and cefoperazone. The self-sorting components of MLp "concentrated island-liked clusters" on the surface of NCefoTs promote the efficient penetration of NCefoTs through the outer membrane. Triggered by the DsbA, the precisely spatiotemporal engineered NCefoTs transform to nanofibers in situ and further significantly enhance the inhibition of DsbA. The hydrolytic activity of ß-lactamase and the motility function of flagella are thereby impeded, confirming the efficacy of NCefoTs in restoring susceptibility to antibiotics and inhibiting infection dissemination. By these synergistic effects of NCefoTs, the minimum inhibitory concentration of antibiotics decreases from over 300 µM to 1.56 µM for clinically isolated E. coli MDR. The survival rate of sepsis-inflicted mice is significantly enhanced from 0% to 92% upon encapsulation of cefoperazone in NCefoTs, which rapidly eliminates invading pathogens and mitigates inflammation. The universally applicable delivery system, based on an "on demands" strategy, presents a promising prospect for undruggable antibiotic targets in the periplasm to combat MDR bacteria.

Complete mitochondrial genomes of the slugs Deroceraslaeve (Agriolimacidae) and Ambigolimaxvalentianus (Limacidae) provide insights into the phylogeny of Stylommatophora (Mollusca, Gastropoda).

In this study, we sequenced two complete mitogenomes from Deroceraslaeve and Ambigolimaxvalentianus. The mitogenome of Ambigolimaxvalentianus represented the first such data from the family Limacidae. The lengths of the mitogenomes of Deroceraslaeve and Ambigolimaxvalentianus were 14,773 bp and 15,195 bp, respectively. The entire set of 37 mitochondrial genes were identified for both mitogenomes. Compared with the mitogenome of Achatinafulica, the trnP_trnA tRNA cluster was rearranged in both Deroceraslaeve and Ambigolimaxvalentianus. The secondary structures of tRNA and rRNA genes for the two species were predicted. Phylogenetic analyses based on amino acid sequences supported (1) monophyly of Stylommatophora, (2) division of Stylommatophora into the 'achatinoid' clade (i.e., the suborder Achatinina) and the 'non-achatinoid' clade (i.e., the suborder Helicina), (3) placement of the Orthurethra in the 'non-achatinoid' clade, and (4) monophyly of each of the superfamilies Helicoidea, Urocoptoidea, Succineoidea, Arionoidea, Pupilloidea and Limacoidea. The exemplars of Helicidae, Philomycidae and Achatinellidae displayed many more mitochondrial gene rearrangements than other species of Stylommatophora.

Simultaneous effect of different chromatographic conditions on the chromatographic retention of pentapeptide derivatives (HGRFG and NPNPT).

Introduction: Oligopeptides exhibit great prospects for clinical application and its separation is of great importance in new drug development. Methods: To accurately predict the retention of pentapeptides with analogous structures in chromatography, the retention times of 57 pentapeptide derivatives in seven buffers at three temperatures and four mobile phase compositions were measured via reversed-phase high-performance liquid chromatography. The parameters ( k H A , k A , and p K a ) of the acid-base equilibrium were obtained by fitting the data corresponding to a sigmoidal function. We then studied the dependence of these parameters on the temperature (T), organic modifier composition (φ, methanol volume fraction), and polarity ( P m N parameter). Finally, we proposed two six-parameter models with (1) pH and T and (2) pH and φ or P m N as the independent variables. These models were validated for their prediction capacities by linearly fitting the predicted retention factor k-value and the experimental k-value. Results: The results showed that log k H A and log k A exhibited linear relationships with 1 / T , φ or P m N for all pentapeptides, especially for the acid pentapeptides. In the model of pH and T, the correlation coefficient (R2) of the acid pentapeptides was 0.8603, suggesting a certain prediction capability of chromatographic retention. Moreover, in the model of pH and φ or P m N , the R2 values of the acid and neutral pentapeptides were greater than 0.93, and the average root mean squared error was approximately 0.3, indicating that the k-values could be effectively predicted. Discussion: In summary, the two six-parameter models were appropriate to characterize the chromatographic retention of amphoteric compounds, especially the acid or neutral pentapeptides, and could predict the chromatographic retention of pentapeptide compounds.

CRISPR/Cas9-Mediated Mutagenesis of Antennapedia in Spodoptera frugiperda.

The homeotic gene Antennapedia (Antp) has been identified as playing a pivotal role in the morphogenesis of the thorax and wings across various insect species. Leveraging insights from previous studies, the functional characterization of Antp in S. frugiperda was undertaken using RT-qPCR and the CRISPR/Cas9 genome-editing system. Phylogenetic analyses indicate that Antp shares a high degree of sequence homology among Lepidoptera species. The expression profile of SfAntp was detected by RT-qPCR. The results showed that SfAntp was expressed in the whole growth cycle of S. frugiperda, the expression level was the highest in the egg stage, and the expression level was higher from 12 h to 48 h. Tissue-specific expression profiling demonstrated that SfAntp was most abundantly expressed in the thoracic segments and legs. To functionally disrupt SfAntp, two sgRNA sites were designed at the first exon of SfAntp and the gene was knocked out by CRISPR/Cas9 via microinjection. The results showed that the deletion of SfAntp produced a mutant phenotype of thoracic fusion, thoracic leg defect, leg-like protrusions between the head and thoracic segments and pupation deformity. In addition, deletion of SfAntp resulted in high embryo mortality. Through DNA sequencing, it was found that the target site of the SfAntp mutant had different degrees of frameshift mutations, indicating that the mutant phenotype was indeed caused by the knockout of SfAntp.

BDNF gene hydroxymethylation in hippocampus related to neuroinflammation-induced depression-like behaviors in mice.

BACKGROUND: Neuroinflammation is a multifactorial condition related to glial cells and neurons activation, and it is implicated in CNS disorders including depression. BDNF is a crucial molecule that related to the pathology of depression, and it is the target of DNA methylation. DNA hydroxymethylation, an active demethylation process can convert 5-mC to 5-hmC by Tets catalyzation to regulate gene transcription. The regulatory function for BDNF gene in response to neuroinflammation remains poorly understood. METHODS: Neuroinflammation and depressive-like behaviors were induced by lipopolysaccharide (LPS) administration in mice. The microglial activation and cellular 5-hmC localization in the hippocampus were confirmed by immunostaining. The transcripts of Tets and BDNF were examined by qPCR method. The global 5-hmC levels and enrichment of 5-hmC in BDNF gene in the hippocampus were analyzed using dot bolt and hMeDIP-sequencing analysis. RESULTS: LPS administration induced a spectrum of depression-like behaviors (including behavioral despair and anhedonia) and increased expression of Iba-1, a marker for microglia activation, in hippocampus, demonstrating that LPS treatment cloud provide stable model of neuroinflammation with depressive-like behaviors as expected. Our results showed that Tet1, Tet2 and Tet3 mRNA expressions and consequent global 5-hmC levels were significantly decreased in the hippocampus of LPS group compared to saline group. We also demonstrated that 5-hmC fluorescence in the hippocampus located in excitatory neurons identified by CaMK II immunostaining. Furthermore, we demonstrated that the enrichment of 5-hmC in BDNF gene was decreased and corresponding BDNF mRNA was down-regulated in the hippocampus in LPS group compared to saline group. CONCLUSION: Neuroinflammation-triggered aberrant BDNF gene hydroxymethylation in the hippocampus is an important epigenetic element that relates with depression-like behaviors.

Bioactivity of the DMI fungicide mefentrifluconazole against Sclerotium rolfsii, the causal agent of peanut southern blight.

BACKGROUND: Sclerotium rolfsii, the causal agent of peanut southern blight, has become increasingly prevalent and harmful in China, causing serious economic losses to the peanut industry. To effectively manage peanut southern blight, this study evaluated the bioactivity of the new-generation sterol demethylation inhibitor (DMI) fungicide mefentrifluconazole against peanut S. rolfsii. RESULTS: In this study, the DMI fungicide mefentrifluconazole exhibited excellent inhibitory activity against the mycelial growth of S. rolfsii, with a mean EC50 value of 0.21 ± 0.11 mg L-1 and a range of 0.02 to 0.55 mg L-1 for 261 isolates collected from Hebei, Henan and Shandong provinces. Mefentrifluconazole significantly reduced the biomass of mycelia and affected the morphology of hyphae. Although sclerotia were more tolerant to mefentrifluconazole than mycelial growth, mefentrifluconazole greatly inhibited the formation and germination of sclerotia. In addition, sclerotia produced by mefentrifluconazole-treated mycelia were deficient in nutrients (e.g., protein, carbohydrate and lipid). These results indicated that mefentrifluconazole may reduce the population of S. rolfsii in the following year. In greenhouse experiments, mefentrifluconazole showed control efficacy and good persistence against peanut S. rolfsii. The preventative and curative activities of mefentrifluconazole at 200 mg L-1 against southern blight still reached 95.36% and 60.94%, respectively, after 9 days of application. No correlation was observed for the sensitivity of S. rolfsii to mefentrifluconazole and the tested DMI, quinone outside inhibitor and succinate dehydrogenase inhibitor fungicides. CONCLUSION: All data indicated that mefentrifluconazole could provide favorable control efficacy against S. rolfsii from peanuts and reduce the infection and population of S. rolfsii in the following year. © 2023 Society of Chemical Industry.

Lymphocyte Membrane- and 12p1-Dual-Functionalized Nanoparticles for Free HIV-1 Trapping and Precise siRNA Delivery into HIV-1-Infected Cells.

Despite the success of small interfering RNA (siRNA) in clinical settings and its potential value in human immunodeficiency virus (HIV) therapy, the rapid clearance and absence of precise delivery to target cells still hinder the therapeutic effect of siRNA. Herein, a new system, which can escape immune recognition, has HIV-1 neutralizing capacity, and the ability to deliver siRNA specifically into HIV-1-infected cells, is constructed by functionalizing siRNA delivery lipid nanoparticles with the lymphocyte membrane and 12p1. The constructed system is shown to escape uptake by the mononuclear phagocyte system. The constructed system exhibits strong binding ability with gp120, thus displaying distinguished neutralizing breadth and potency. The constructed system neutralizes all tested HIV-1 pseudotyped viruses with a geometric mean 80% inhibitory concentration (IC80) of 29.75 µg mL-1 and inhibits X4-tropic HIV-1 with an IC80 of 64.20 µg mL-1 , and R5-tropic HIV-1 with an IC80 of 16.39 µg mL-1 . The new system also specifically delivers siRNA into the cytoplasm of HIV-1-infected cells and exhibits evident gene silencing of tat and rev. Therefore, this new system can neutralize HIV-1 and deliver siRNA selectively into HIV-1-infected cells and may be a promising therapeutic candidate for the precise therapy of HIV.