Mitochondrial biogenesis disorder and oxidative damage promote refractory apical periodontitis in rat and human.

AIM: To elucidate whether mitochondrial biogenesis disorder and damage from oxidative stress promote refractory apical periodontitis (RAP) in rat and human. METHODOLOGY: Twenty Enterococcus faecalis-induced RAPs were established in the maxillary first molars of male Wistar rats. Concurrently, 12 periapical lesion specimens from patients presenting with RAP were obtained by apicoectomy. Radiographic examination and histologic analysis were conducted to evaluate periapical bone tissue destruction and morphological changes. The expression of key regulators of mitochondrial biogenesis, PGC-1α and Nrf2, were detected by immunohistochemistry and double immunofluorescence staining, Western blot and real-time PCR were also assayed. Mitochondrial ROS (mtROS) was identified by MitoSOX staining. Mitochondrial function was detected by the quantification of ATP production, mitochondrial DNA (mtDNA) copy number and activities of mitochondrial respiratory chain complexes. Furthermore, mitochondrial oxidative stress was evaluated by the determination of 3-nitrotyrosine (3-NT), 4-hydroxy-2-nonenal (4-HNE) and 8-hydroxy-deoxyguanosine (8-OHdG) expression levels, as well as malondialdehyde (MDA) expression and antioxidant capacity. Student's t-test was performed to determine significance between the groups; p < .05 was considered significant. RESULTS: In the maxilla, significantly more bone resorption, greater number of periapical apoptotic cells and Tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells were observed in the RAP group compared with the control group (p < .01). PGC-1α and Nrf2 were significantly reduced in rat and human RAP lesions compared to the control group (p < .01) at both the mRNA and protein levels. Double immunofluorescence analysis of PGC-1α or Nrf2 with TOMM20 also indicated that mitochondrial biogenesis was impaired in RAP group (p < .01). Additionally, mitochondrial dysfunction was observed in RAP group, as reflected by increased mtROS, decreased ATP production, reduced mtDNA copy number and complexes of the mitochondrial respiratory chain. Finally, the expression levels of mitochondrial oxidative stress markers, 3-NT, 4-HNE and 8-OHdG, were significantly increased in the RAP group (p < .01). Consistent with this, systemic oxidative damage was also present in the progression of RAP, including increased MDA expression and decreased antioxidant activity (p < .01). CONCLUSIONS: Mitochondrial biogenesis disorder and damage from oxidative stress contribute to the development of RAP.

The development of an egg-soaking method for delivering dsRNAs into spider mites.

Recently, the first sprayable RNAi biopesticide, Ledprona, against the Colorado potato beetle, Leptinotarsa decemlineata, has been registered at the United States Environmental Protection Agency. Spider mites (Acari: Tetranychidae), a group of destructive agricultural and horticultural pests, are notorious for rapid development of insecticide/acaricide resistance. The management options, on the other hand, are extremely limited. RNAi-based biopesticides offer a promising control alternative to address this emerging issue. In this study, we i) developed an egg-soaking dsRNA delivery method; ii) evaluated the factors influencing RNAi efficiency, and finally iii) investigated the potential mode of entry of this newly developed egg-soaking RNAi method. In comparison to other dsRNA delivery methods, egg-soaking method was the most efficient, convenient/practical, and cost-effective method for delivering dsRNAs into spider mites. RNAi efficiency of this RNAi method was affected by target genes, dsRNA concentration, developmental stages, and mite species. In general, the hawthorn spider mite, Amphitetranychus viennensis, is more sensitive to RNAi than the two-spotted spider mite, Tetranychus urticae, and both of them have dose-dependent RNAi effect. For different life stages, egg and larvae are the most sensitive life stages to dsRNAs. For different target genes, there is no apparent association between the suppression level and the resultant phenotype. Finally, we demonstrated that this egg-soaking RNAi method acts as both stomach and contact toxicity. Our combined results demonstrate the effectiveness of a topically applied dsRNA delivery method, and the potential of a spray induced gene silencing (SIGS) method as a control alternative for spider mites.

A bioenergy-focused versus a reforestation-focused mitigation pathway yields disparate carbon storage and climate responses.

Limiting global warming to 2 °C requires urgent action on land-based mitigation. This study evaluates the biogeochemical and biogeophysical implications of two alternative land-based mitigation scenarios that aim to achieve the same radiative forcing. One scenario is primarily driven by bioenergy expansion (SSP226Lu-BIOCROP), while the other involves re/afforestation (SSP126Lu-REFOREST). We find that overall, SSP126Lu-REFOREST is a more efficient strategy for removing CO2 from the atmosphere by 2100, resulting in a net carbon sink of 242 ~ 483 PgC with smaller uncertainties compared to SSP226Lu-BIOCROP, which exhibits a wider range of -78 ~ 621 PgC. However, SSP126Lu-REFOREST leads to a relatively warmer planetary climate than SSP226Lu-BIOCROP, and this relative warming can be intensified in certain re/afforested regions where local climates are not favorable for tree growth. Despite the cooling effect on a global scale, SSP226Lu-BIOCROP reshuffles regional warming hotspots, amplifying summer temperatures in vulnerable tropical regions such as Central Africa and Southeast Asia. Our findings highlight the need for strategic land use planning to identify suitable regions for re/afforestation and bioenergy expansion, thereby improving the likelihood of achieving the intended climate mitigation outcomes.

To explore the effect of kaempferol on non-small cell lung cancer based on network pharmacology and molecular docking.

Non-small cell lung cancer (NSCLC) is a common pathological type of lung cancer, which has a serious impact on human life, health, psychology and life. At present, chemotherapy, targeted therapy and other methods commonly used in clinic are prone to drug resistance and toxic side effects. Natural extracts of traditional Chinese medicine (TCM) have attracted wide attention in cancer treatment because of their small toxic and side effects. Kaempferol is a flavonoid from natural plants, which has been proved to have anticancer properties in many cancers such as lung cancer, but the exact molecular mechanism is still unclear. Therefore, on the basis of in vitro experiments, we used network pharmacology and molecular docking methods to study the potential mechanism of kaempferol in the treatment of non-small cell lung cancer. The target of kaempferol was obtained from the public database (PharmMapper, Swiss target prediction), and the target of non-small cell lung cancer was obtained from the disease database (Genecards and TTD). At the same time, we collected gene chips GSE32863 and GSE75037 in conjunction with GEO database to obtain differential genes. By drawing Venn diagram, we get the intersection target of kaempferol and NSCLC. Through enrichment analysis, PI3K/AKT is identified as the possible key signal pathway. PIK3R1, AKT1, EGFR and IGF1R were selected as key targets by topological analysis and molecular docking, and the four key genes were further verified by analyzing the gene and protein expression of key targets. These findings provide a direction for further research of kaempferol in the treatment of NSCLC.

Surfactants Improving the Wetting Behavior and Adhesion Mechanism of Pesticide Dilution Droplets on Jujube Leaf Surfaces.

Fruit tree leaves have different chemical compositions and diverse wax layer structures that result in different patterns of wetting and pesticide solution spreading on their surface. Fruit development is a time when pests and diseases occur, during which a large number of pesticides are needed. The wetting and diffusion properties of pesticide droplets on fruit tree leaves were relatively poor. To solve this problem, the wetting characteristics of leaf surfaces with different surfactants were studied. The contact angle, surface tension, adhesive tension, adhesion work, and solid-liquid interfacial tension of five surfactant solution droplets on jujube leaf surfaces during fruit growth were studied by the sessile drop method. C12E5 and Triton X-100 have the best wetting effects. Two surfactants were added to a 3% beta-cyfluthrin emulsion in water, and field efficacy tests were carried out on peach fruit moths in a jujube orchard at different dilutions. The control effect is as high as 90%. During the initial stage when the concentration is low, due to the surface roughness of the leaves, the surfactant molecules adsorbed at the gas-liquid and solid-liquid interfaces reach an equilibrium, and the contact angle on the leaf surface changes slightly. With increasing surfactant concentration, the pinning effect in the spatial structure on the leaf surface is overcome by liquid droplets, thereby significantly decreasing the contact angle. When the concentration is further increased, the surfactant molecules form a saturated adsorption layer on the leaf surface. Due to the existence of a precursor water film in the droplets, surfactant molecules on the interface continuously move to the water film on the surface of jujube tree leaves, thus causing interactions between the droplets and the leaves. The conclusion of this study provides theoretical guidance for the wettability and adhesion of pesticides on jujube leaves, so as to achieve the purpose of reducing pesticide use and improving pesticide efficacy.

Target gene selection for RNAi-based biopesticides against the hawthorn spider mite, Amphitetranychus viennensis (Acari: Tetranychidae).

BACKGROUND: Recently, RNA interference (RNAi)-based biopesticide, a species-specific pest control alternative, has been deregulated and commercialized in the US and Canada. The hawthorn spider mite, Amphitetranychus viennensis Zacher, is a major pest for rosaceous plants, which has been controlled primarily by synthetic pesticides. To address the emerging resistance issues in A. viennensis, we initiated a project to develop RNAi-based biopesticides. RESULTS: In this study, we (i) developed a dietary RNAi system for A. viennensis using leaf disc, (ii) assessed the suitability of multiple control genes to distinguish sequence-specific silencing from non-specific effects within this RNAi system, and (iii) screened for the target gene candidates. As a result, ß-Glucuronidase (GUS), an enzyme derived from E. coli and a broadly used reporter for plants is the appropriate control for A. viennensis RNAi, while green fluorescent protein (GFP), is not suitable due to its significantly higher mortality than the other controls. For target gene screening, suppression was confirmed for all the candidates, including two housekeeping genes (Vacuolar-type H + -ATPase subunit A (V-ATPase A) and Glyceraldehyde 3-phosphate dehydrogenase, (GAPDH)), and three genes associated with development (ATP-dependent RNA Helicase DDX3Y (Belle), CREB-binding protein (CBP), and Farnesoic acid O-methyltransferase (FaMet)). Knocking down of V-ATPase A resulted in the highest mortality (~ 90%) and reduced fecundity (over 90%) than other candidates. As for the genes associated with development, suppression of Belle and CBP, led to approximately 65% mortality, as well as 86% and 40% reduction in fecundity, respectively. Silencing of FaMet, however, had negligible biological impacts on A. viennensis. CONCLUSION: The combined efforts not only establish an effective dsRNA delivery method, but also provide potential target genes for RNAi-based biopesticides against A. viennensis, a devastating invasive pest for fruit trees and woody ornamental plants throughout Asia and Europe. © 2023 Society of Chemical Industry.

Energy Utilization and Flight Muscle Development in Chrysoperla sinica (Neuroptera: Chrysopidae) Female Adults: Relationship With Age and Flight Behavior.

We investigated the accumulation of energy substances, dynamics of flight muscle development, changes in energy substances accumulation, and flight muscle structure after flight activities in female adults of the green lacewing Chrysoperla sinica (Tjeder), a common natural enemy of various insect pests in China. Virgin individuals were chosen at 24, 72, and 120 h after eclosion for energy substance determination and flight muscle observation in this study. Individuals with strong flight ability at 72 h after eclosion were selected for tethered flight assays, followed by detection of energy substances, and flight muscle observation. The results showed that virgin female adults had the highest fat content 72 h after eclosion. Sarcomere length and myofibril diameter changed significantly with age, with the lowest at 24 h after eclosion. With an increase in flight distance, the fat and glycogen contents, sarcomere length, and volume fraction of the transverse tubular system (T-system) decreased and myofibril diameters increased. The volume fraction of the mitochondria did not significantly change, but the structure of the mitochondrial membrane was destroyed, inclusions were reduced, and cavities appeared. The reserves of energy substances, especially lipids, are closely related to the flight ability of C. sinica. The observational results of both flight muscle structure and morphology of mitochondria build a strong relationship with flight behavior. This research should help reveal the regulatory mechanism of flight activity of C. sinica.

Warm pool ocean heat content regulates ocean-continent moisture transport.

The Indo-Pacific Warm Pool (IPWP) exerts a dominant role in global climate by releasing huge amounts of water vapour and latent heat to the atmosphere and modulating upper ocean heat content (OHC), which has been implicated in modern climate change1. The long-term variations of IPWP OHC and their effect on monsoonal hydroclimate are, however, not fully explored. Here, by combining geochemical proxies and transient climate simulations, we show that changes of IPWP upper (0-200 m) OHC over the past 360,000 years exhibit dominant precession and weaker obliquity cycles and follow changes in meridional insolation gradients, and that only 30%-40% of the deglacial increases are related to changes in ice volume. On the precessional band, higher upper OHC correlates with oxygen isotope enrichments in IPWP surface water and concomitant depletion in East Asian precipitation as recorded in Chinese speleothems. Using an isotope-enabled air-sea coupled model, we suggest that on precessional timescales, variations in IPWP upper OHC, more than surface temperature, act to amplify the ocean-continent hydrological cycle via the convergence of moisture and latent heat. From an energetic viewpoint, the coupling of upper OHC and monsoon variations, both coordinated by insolation changes on orbital timescales, is critical for regulating the global hydroclimate.

Atmospheric forcing dominates winter Barents-Kara sea ice variability on interannual to decadal time scales.

The last two decades have seen a dramatic decline and strong year-to-year variability in Arctic winter sea ice, especially in the Barents-Kara Sea (BKS), changes that have been linked to extreme midlatitude weather and climate. It has been suggested that these changes in winter sea ice arise largely from a combined effect of oceanic and atmospheric processes, but the relative importance of these processes is not well established. Here, we explore the role of atmospheric circulation patterns on BKS winter sea ice variability and trends using observations and climate model simulations. We find that BKS winter sea ice variability is primarily driven by a strong anticyclonic anomaly over the region, which explains more than 50% of the interannual variability in BKS sea-ice concentration (SIC). Recent intensification of the anticyclonic anomaly has warmed and moistened the lower atmosphere in the BKS by poleward transport of moist-static energy and local processes, resulting in an increase in downwelling longwave radiation. Our results demonstrate that the observed BKS winter sea-ice variability is primarily driven by atmospheric, rather than oceanic, processes and suggest a persistent role of atmospheric forcing in future Arctic winter sea ice loss.

Effects of Imidacloprid Applied Alone or in Combination With Organosilicone Surfactants on Biological Traits and Predatory Feeding of Chrysoperla nipponensis (Neuroptera: Chrysopidae).

Organosilicone adjuvants are widely used to increase insecticide application on targeted surfaces. In this work, our aim was to investigate side effects of imidacloprid treatment, either applied alone or in combination with organosilicone compounds, against Chrysoperla nipponensis, an important predator of aphids. Four types of organosilicones were mixed with imidacloprid at different concentrations. The toxicity of the mixture to C. nipponensis was measured under laboratory conditions. The LC50 and LC30 of imidacloprid applied alone and in combination with 0.05% organosilicone were determined. Imidacloprid (LC30) applied alone or in combination with 0.05% organosilicone was used to treat second instar larvae of C. nipponensis; thereafter, its effects on the growth, development, longevity, reproduction, and predatory ability of C. nipponensis were evaluated The results demonstrated that the organosilicone Silwet L-77 reduced the LC50 and LC30 of imidacloprid to 6.09 (95% CI: 2.31-9.42) and 10.95 mg/L (95% CI: 8.16-13.63), respectively, and enhanced imidacloprid toxicity to C. nipponensis, as reflected by the resulting extension of the growth and developmental period, reduction in female longevity, and inhibition of reproduction. When applied alone or in combination with an organosilicone, imidacloprid reduced the consumption of Corcyra cephalonica eggs by C. nipponensis. The functional response of C. nipponensis treated with imidacloprid alone or in combination with organosilicone was type II. Concomitantly, the attack rate was reduced and the handling time of prey increased.

Acceleration of western Arctic sea ice loss linked to the Pacific North American pattern.

Recent rapid Arctic sea-ice reduction has been well documented in observations, reconstructions and model simulations. However, the rate of sea ice loss is highly variable in both time and space. The western Arctic has seen the fastest sea-ice decline, with substantial interannual and decadal variability, but the underlying mechanism remains unclear. Here we demonstrate, through both observations and model simulations, that the Pacific North American (PNA) pattern is an important driver of western Arctic sea-ice variability, accounting for more than 25% of the interannual variance. Our results suggest that the recent persistent positive PNA pattern has led to increased heat and moisture fluxes from local processes and from advection of North Pacific airmasses into the western Arctic. These changes have increased lower-tropospheric temperature, humidity and downwelling longwave radiation in the western Arctic, accelerating sea-ice decline. Our results indicate that the PNA pattern is important for projections of Arctic climate changes, and that greenhouse warming and the resultant persistent positive PNA trend is likely to increase Arctic sea-ice loss.

Analysis of Differentially Expressed Genes of Chrysoperla sinica Related to Flight Capacity by Transcriptome.

The lacewing Chrysoperla sinica (Tjeder) is a common natural enemy of many insect pests in China and is frequently employed for biological control programs. Adults make migratory flights after emergence, which reduces their effectiveness as biological control agents. Previously, we proved that 2-d-old unmated females exhibited significantly stronger flight ability than 3-d-old ones. Meanwhile, 3-d-old unmated adults flew significantly longer distances than mated ones. In this study, Illumina RNA sequencing was performed to characterize differentially expressed genes (DEGs) between virgin and mated adults of different ages in a single female strain of C. sinica. In total, 713,563,726 clean reads were obtained and de novo assembled into 109,165 unigenes with an average length of 847 bp (N50 of 1,754 bp), among which 4,382 (4.01%) unigenes matched known proteins. Based on these annotations, many putative transcripts were related to C. sinica's flight capacity and muscle structure, energy supply, growth, development, environmental adaptability, and metabolism of nutritional components and bioactive components. In addition, the differential expression of transcripts between different ages and mating status were analyzed, and DEGs participating in flight capacity and muscles were detected, including glutathione hydrolase, NAD-specific glutamate dehydrogenase, aminopeptidase, and acidic amino acid decarboxylase. The DEGs with functions associated with flight capacity and muscles exhibited higher transcript levels for younger (2 d--old) virgins. This comprehensive C. sinica transcriptomic data provide a foundation for a better understanding of the molecular mechanisms underlying the flight capacity to meet the physiological demands of flight muscles in C. sinica.

Selection of Reference Genes for RT-qPCR Analysis Under Extrinsic Conditions in the Hawthorn Spider Mite, Amphitetranychus viennensis.

Hawthorn spider mite, Amphitetranychus viennensis Zacher, is an economically important arthropod pest for fruit trees and woody ornamental plants. Extensive and repetitive use of synthetic acaricides has led to the development of resistance in A. viennensis. To understand the molecular basis of pesticide resistance, and to develop genetic-based control alternatives (e.g., RNAi-based biopesticides), a standardized protocol for real-time quantitative reverse transcription PCR (RT-qPCR) is needed. In the proceeding phase of this research, we screened for the internal references for RT-qPCR analysis from a pool of A. viennensis housekeeping genes under the intrinsic conditions, including developmental stage, sex, and diapause. Here, we continued our efforts to search for the reference genes under an array of extrinsic conditions, including temperature, humidity, photoperiod, host plant, and dietary RNAi. The stability of these candidate reference genes was investigated using geNorm, NormFinder, BestKeeper, and ΔCt method, respectively. Finally, RefFinder, a statistical platform integrating all four algorisms, provided a comprehensive list of genes for each extrinsic condition: (1) EF1A, α-tubulin and Actin3 were the best candidates for temperature, (2) GAPDH, 18S, and Actin3 were the most stable genes for humidity, (3) V-ATPase B, Actin3, and 18S were the top reference genes for photoperiod, (4) GAPDH, V-ATPase B, and α-tubulin were recommended for host plants, and (5) GAPDH, V-ATPase B, and RPS9 were the top choices for dietary RNAi. Overall, V-ATPase B, GAPDH, and Actin3 were the most commonly selected reference genes in A. viennensis regardless of the experimental conditions, including both intrinsic and extrinsic. Information present here lays the foundation for the genomic and functional genomic research in A. viennensis.

Wetting and adhesion behavior on apple tree leaf surface by adding different surfactants.

OBJECTIVE: This study was conducted to investigate the wetting behavior of different surfactant solutions on the leaf surfaces of apple during the fruit formation stage. METHODS: Five surfactants, including C12E5, Tween-20, Triton X-100, DTAB, and SDS were evaluated in this study. The contact angle, surface tension, adhesion tension, work of adhesion, and solid-liquid interface tension of droplets on the leaf surface were determined by the drop method. RESULTS: The results showed that the nonionic surfactants C12E5 and Triton X-100 had better wetting effects than other surfactants. Moreover, when the concentration of C12E5 and Triton X-100 was 1 × 10-3 mol/L, the leaves reached a completely wet state. Toxicity measurement showed that the incubation rate of Carposina niponensis eggs decreased gradually with increasing content of C12E5 or Triton X-100. Additionally, field efficacy analysis showed that adding C12E5 or Triton X-100 significantly improved the beta-cyfluthrin 3% water emulsion (EW) against C. niponensis. CONCLUSIONS: These results indicate that the surfactants C12E5 and Triton X-100 can significantly improve pesticide application, which will be helpful for reducing pesticide use and developing new pesticides.

Selection of Reference Genes for RT-qPCR Analysis Under Intrinsic Conditions in the Hawthorn Spider Mite, Amphitetranychus viennensis (Acarina: Tetranychidae).

Hawthorn spider mite, Amphitetranychus viennensis Zacher, is one of the most devastating pests of deciduous fruit trees. The overall goal of this research is to develop a standardized protocol for real-time quantitative reverse transcription PCR (RT-qPCR) analysis in A. viennensis following the MIQE (minimum information for publication of Quantitative real time PCR experiments) guidelines. Based on the previous knowledge, we hypothesized that internal references for RT-qPCR analysis reside in housekeeping genes (HKGs). To test this hypothesis, we examined the stability of nine HKGs from A. viennensis, including 18S ribosomal RNA (18S), 28S ribosomal RNA (28S), Elongation factor 1-alpha (EF1A), Actin3, V-ATP vacuolar-type H+-ATPase (V-ATPase), α-tubulin (α-tubulin), Ribosomal protein L13 (RPL13), 40S ribosomal protein S9 (RPS9), and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The expression profile of these candidates under intrinsic conditions was evaluated by a panel of computational programs, including geNorm, Normfinder, BestKeeper, and ΔCt method. Based on RefFinder, a comprehensive software integrating all four above-mentioned algorithms, V-ATPase, Actin3, and GAPDH are the top three reference genes, which are stably expressed across all the intrinsic conditions, including developmental stage, sex, and diapause. In addition, we compared reference genes recommended for different developmental stages among the nine cell-content feeding arthropods, including four spider mites, A. viennensis, Tetranychus urticae, Tetranychus cinnabarinus, and Panonychus citri, and five hemipterans, Myzus persicae, Aphis gossypii, Toxoptera citricida, Lipaphis erysimi, and Sogatella furcifera. Not surprisingly, rRNAs and ribosomal proteins, the most abundant RNA species, is the top choice, and follows by EF1A, Actin, GAPDH, and tubulin. Information present here lays the foundation for the genomic and functional genomic research in cell-content feeding arthropods in general and A. viennensis in particular.

The Combinations Chitosan-Pam3CSK4 and Chitosan-Monophosphoryl Lipid A: Promising Immune-Enhancing Adjuvants for Anticaries Vaccine PAc.

We previously demonstrated that recombinant protein PAc could be administered as an anticaries vaccine. However, the relatively weak immunogenicity of PAc limits its application. In the present study, we investigated the effect of two adjuvant combinations of chitosan plus Pam3CSK4 (chitosan-Pam3CSK4) and of chitosan plus monophosphoryl lipid A (chitosan-MPL) in the immune responses to the PAc protein in vivo and in vitro PAc-chitosan-Pam3CSK4 or PAc-chitosan-MPL promoted significantly higher PAc-specific antibody titers in serum and saliva, inhibited Streptococcus mutans colonization onto the tooth surfaces, and endowed better protection effect with significantly less caries activities than PAc alone. Chitosan-Pam3CSK4 and chitosan-MPL showed no statistically significant differences. In conclusion, our study demonstrated that the chitosan-Pam3CSK4 and chitosan-MPL combinations are promising for anticaries vaccine development.

Identification of FimH derivatives as adjuvant vaccinated with PAc that enhance protection against Streptcoccus mutans colonization.

FimH is the adhesin of type I fimbriae expressed on Escherichia coli that can mediate specific adherence to host cells. High binding mutations in FimH are related to the adaptive evolution of bacteria. However, additional roles that these allelic variations may play remain elusive. To investigate novel biological functions of the mutations in FimH, we introduced four different variants of FimH by incorporating single amino acid substitutions at specific sites, namely A25P, G73R, A106, and T158P, respectively. In this study, adjuvant potential of FimH variants was evaluated by investigating their ability to trigger innate immune response to DC2.4 and adaptive immunity to improve immunological characteristics. The data revealed that purified A106 and T158P up-regulated the expression of co-stimulatory molecules critically involved in DC2.4 activation by interaction with TLR4, whereas A25P and G73R did not induce the phenotypic maturation of DC2.4. Besides, the culture of DC2.4 with A106 and T158P enhanced the release of cytokines and protein phagocytosis. When formulated with PAc, T158P elicited more robust PAc-specific IgG and IgA antibody responses compared to PBS, PAc and PAc+K12 groups and inhibited bacteria colonization. Collectively, the results confirmed that the T158P mutation located around the inter-domain interface of the protein induced a specific enhancement effect on adjuvant characteristics.

FimH as a mucosal adjuvant enhances persistent antibody response and protective efficacy of the anti-caries vaccine.

OBJECTIVE: To investigate whether the recombinant FimH-S.T protein could modulate immune response to anti-caries vaccine in vitro and in vivo. DESIGN: Recombinant FimH protein derived from Salmonella was constructed and purified. The expression of dendritic cell maturation markers and cytokines release were performed by flow cytometry, Real-time PCR and ELISA. In addition, BALB/c mice were administered with anti-caries PAc vaccine plus FimH-S.T, antibody responses were evaluated by ELISA. Splenocytes of immunized mice were detected for their proliferative ability in response to in vitro retreatment with PAc antigen by flow cytometry. Caries protection against dental caries formation was also investigated. RESULTS: The purified FimH-S.T induced phenotypic maturation of DC2.4 by up-regulating the expression of costimulatory molecules and MHC II, provoked the production and secretion of cytokines via TLR4-dependent signaling pathway in vitro. Furthermore, the mice immunized with the mixture of FimH-S.T and PAc significantly enhanced the PAc-specific antibodies in the serum along with saliva and promoted splenocyte proliferation. Our results also confirmed that PAc+FimH-S.T decreased the caries lesions formation which provided high protective efficacy against dental caries. CONCLUSION: Our study demonstrates that recombinant FimH-S.T could enhance specific IgA responses and protection of anti-caries vaccine, possessing mucosal adjuvant ability by activating DC2.4 via TLR4 signaling pathway.

HDAC3-mediated silencing of miR-451 decreases chemosensitivity of patients with metastatic castration-resistant prostate cancer by targeting NEDD9.

BACKGROUND: Treatment of metastatic castration-resistant prostate cancer (mCRPC) with docetaxel often fails due to the emergence of chemoresistance. Thus, restoring chemosensitivity to docetaxel-based therapies remains a challenge in mCRPC treatment. METHODS: microRNA (miR)-451 expression was measured in docetaxel-treated prostate cancer cells and tumor tissues by quantitative reverse-transcription polymerase chain reaction . Cell-counting kit 8 assay was performed to determine docetaxel chemoresistance. Neural-precursor-cell-expressed developmentally downregulated protein 9 (NEDD9) was identified as a novel target of miR-451 by dual-luciferase reporter system. Chromatin immunoprecipitation and co-immunoprecipitation assay were performed to confirm that histone deacetylase 3 (HDAC3)/Sp1 (a highly evolutionarily conserved transcription factor) interacted with the Sp1 binding sites in miR-451 promoter. RESULTS: miR-451 was found to be silenced in docetaxel-treated prostate cancer cells and mCRPC tissues. Low miR-451 expression was closely associated with a high Gleason score, high Eastern Cooperative Oncology Group performance status score, visceral metastasis and poor prognosis. Low expression of miR-451 was significantly correlated with short progression-free survival (PFS) and overall survival (OS) according to Kaplan-Meier analysis, and miR-451 was determined to be an independent poor prognostic factor for PFS and OS in mCRPC patients by univariate and multivariate Cox regression analyses. NEDD9 was identified as a new and functional target of miR-451. Restoration of NEDD9 partially reversed the effects of miR-451 on enhancing chemosensitivity of prostate cancer cells. HDAC3 was confirmed to be involved in silencing of miR-451 expression in prostate cancer cells. CONCLUSIONS: The current data revealed a new HDAC3/Sp1/miR-451/NEDD9 signaling axis that regulates the chemosensitivity of prostate cancer cells and represents a novel therapeutic target for chemosensitizing mCRPC.

Wettability of pear leaves from three regions characterized at different stages after flowering using the OWRK method.

BACKGROUND: A better understanding of leaf surface wettability is critical to improve the adhesion of liquid pesticides. Leaf surface wettability is dependent on the property of the liquid as well as the physical and chemical properties of the leaf, which vary with climate and growth stage. The aim of this study was to characterize the wettability of pear leaves from three different climatic regions at different stages after flowering. RESULTS: The contact angles of different test liquids were measured on both adaxial and abaxial pear leaf surfaces and the Owens-Wendt-Rabel-Kaelble (OWRK) method was used to calculate surface free energy (SFE) and its polar and non-polar components. The results demonstrated that the SFE of both the adaxial and abaxial surface of the pear leaf, and the proportion of polar component, increased with increasing time after flowering. At early growth stages, pear leaves were highly hydrophobic, similar to a polytetrafluoroethylene surface, whereas at later growth stages, pear leaves were hydrophobic, more similar to a polymethylmethacrylate surface. Also, the SFE differed with climatic region. Factors influencing these changes are discussed. CONCLUSION: Changes in contact angles and SFE correlated with the change of the leaf surface wettability. Leaves became easier to wet (higher SFE), with an overall increasing polar component to the surface, with increasing age after flowering. As expected, changes in wettability were found in pear leaves at different stages after flowering and in different regions (P < 0.05). Pear leaves from Yuanping were easier to wet than leaves from Yuci and Linyi, and adaxial surfaces were easier to wet than abaxial surfaces. These results provide beneficial information for the application of agrochemicals for improved wetting and spreading behavior. © 2018 Society of Chemical Industry.